Abnormal nuclear envelopes in the striatum and motor deficits in DYT11 myoclonus-dystonia mouse models.

PubMed

Yokoi, Fumiaki; Dang, Mai T; Zhou, Tong; Li, Yuqing

2012-02-15

DYT11 myoclonus-dystonia (M-D) is a movement disorder characterized by myoclonic jerks with dystonic symptoms and caused by mutations in paternally expressed SGCE, which codes for ε-sarcoglycan. Paternally inherited Sgce heterozygous knock-out (KO) mice exhibit motor deficits and spontaneous myoclonus. Abnormal nuclear envelopes have been reported in cellular and mouse models of early-onset DYT1 generalized torsion dystonia; however, the relationship between the abnormal nuclear envelopes and motor symptoms are not clear. Furthermore, it is not known whether abnormal nuclear envelope exists in non-DYT1 dystonia. In the present study, abnormal nuclear envelopes in the striatal medium spiny neurons (MSNs) were found in Sgce KO mice. To analyze whether the loss of ε-sarcoglycan in the striatum alone causes abnormal nuclear envelopes, motor deficits or myoclonus, we produced paternally inherited striatum-specific Sgce conditional KO (Sgce sKO) mice and analyzed their phenotypes. Sgce sKO mice exhibited motor deficits in both beam-walking and accelerated rotarod tests, while they did not exhibit abnormal nuclear envelopes, alteration in locomotion, or myoclonus. The results suggest that the loss of ε-sarcoglycan in the striatum contributes to motor deficits, while it alone does not produce abnormal nuclear envelopes or myoclonus. Development of therapies targeting the striatum to compensate for the loss of ε-sarcoglycan function may rescue the motor deficits in DYT11 M-D patients.

Role of the nuclear envelope in the pathogenesis of age-related bone loss and osteoporosis

PubMed Central

Vidal, Christopher; Bermeo, Sandra; Fatkin, Diane; Duque, Gustavo

2012-01-01

The nuclear envelope is the most important border in the eukaryotic cell. The role of the nuclear envelope in cell differentiation and function is determined by a constant interaction between the elements of the nuclear envelope and the transcriptional regulators involved in signal transcription pathways. Among those components of the nuclear envelope, there is a growing evidence that changes in the expression of A-type lamins, which are essential components of the nuclear lamina, are associated with age-related changes in bone affecting the capacity of differentiation of mesenchymal stem cells into osteoblasts, favoring adipogenesis and affecting the function and survival of the osteocytes. Overall, as A-type lamins are considered as the 'guardians of the soma', these proteins are also essential for the integrity and quality of the bone and pivotal for the longevity of the musculoskeletal system. PMID:23951459

Abnormal nuclear envelopes in the striatum and motor deficits in DYT11 myoclonus-dystonia mouse models

PubMed Central

Yokoi, Fumiaki; Dang, Mai T.; Zhou, Tong; Li, Yuqing

2012-01-01

DYT11 myoclonus-dystonia (M-D) is a movement disorder characterized by myoclonic jerks with dystonic symptoms and caused by mutations in paternally expressed SGCE, which codes for ɛ-sarcoglycan. Paternally inherited Sgce heterozygous knock-out (KO) mice exhibit motor deficits and spontaneous myoclonus. Abnormal nuclear envelopes have been reported in cellular and mouse models of early-onset DYT1 generalized torsion dystonia; however, the relationship between the abnormal nuclear envelopes and motor symptoms are not clear. Furthermore, it is not known whether abnormal nuclear envelope exists in non-DYT1 dystonia. In the present study, abnormal nuclear envelopes in the striatal medium spiny neurons (MSNs) were found in Sgce KO mice. To analyze whether the loss of ɛ-sarcoglycan in the striatum alone causes abnormal nuclear envelopes, motor deficits or myoclonus, we produced paternally inherited striatum-specific Sgce conditional KO (Sgce sKO) mice and analyzed their phenotypes. Sgce sKO mice exhibited motor deficits in both beam-walking and accelerated rotarod tests, while they did not exhibit abnormal nuclear envelopes, alteration in locomotion, or myoclonus. The results suggest that the loss of ɛ-sarcoglycan in the striatum contributes to motor deficits, while it alone does not produce abnormal nuclear envelopes or myoclonus. Development of therapies targeting the striatum to compensate for the loss of ɛ-sarcoglycan function may rescue the motor deficits in DYT11 M-D patients. PMID:22080833

Mechanism of Dissolution of Envelopes of the Extreme Halophile Halobacterium cutirubrum1

PubMed Central

Onishi, H.; Kushner, D. J.

1966-01-01

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 NH4Cl. This lysis did not involve an extensive breakdown of the envelope. No evidence for different sites of Na+, K+, and NH4+ 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. Images PMID:5883109

Nuclear transport of cancer extracellular vesicle-derived biomaterials through nuclear envelope invagination-associated late endosomes.

PubMed

Rappa, Germana; Santos, Mark F; Green, Toni M; Karbanová, Jana; Hassler, Justin; Bai, Yongsheng; Barsky, Sanford H; Corbeil, Denis; Lorico, Aurelio

2017-02-28

Extracellular membrane vesicles (EVs) function as vehicles of intercellular communication, but how the biomaterials they carry reach the target site in recipient cells is an open question. We report that subdomains of Rab7+ late endosomes and nuclear envelope invaginations come together to create a sub-nuclear compartment, where biomaterials associated with CD9+ EVs are delivered. EV-derived biomaterials were also found in the nuclei of host cells. The inhibition of nuclear import and export pathways abrogated the nuclear localization of EV-derived biomaterials or led to their accumulation therein, respectively, suggesting that their translocation is dependent on nuclear pores. Nuclear envelope invagination-associated late endosomes were observed in ex vivo biopsies in both breast carcinoma and associated stromal cells. The transcriptome of stromal cells exposed to cancer cell-derived CD9+ EVs revealed that the regulation of eleven genes, notably those involved in inflammation, relies on the nuclear translocation of EV-derived biomaterials. Our findings uncover a new cellular pathway used by EVs to reach nuclear compartment.

Nuclear Envelope Disruption Involving Host Caspases Plays a Role in the Parvovirus Replication Cycle ▿

PubMed Central

Cohen, Sarah; Marr, Alexandra K.; Garcin, Pierre; Panté, Nelly

2011-01-01

Parvoviruses are small, nonenveloped, single-stranded DNA viruses which replicate in the nucleus of the host cell. We have previously found that early during infection the parvovirus minute virus of mice (MVM) causes small, transient disruptions of the nuclear envelope (NE). We have now investigated the mechanism used by MVM to disrupt the NE. Here we show that the viral phospholipase A2, the only known enzymatic domain on the parvovirus capsid, is not involved in causing NE disruption. Instead, the virus utilizes host cell caspases, which are proteases involved in causing NE breakdown during apoptosis, to facilitate these nuclear membrane disruptions. Studies with pharmacological inhibitors indicate that caspase-3 in particular is involved. A caspase-3 inhibitor prevents nuclear lamin cleavage and NE disruption in MVM-infected mouse fibroblast cells and reduces nuclear entry of MVM capsids and viral gene expression. Caspase-3 is, however, not activated above basal levels in MVM-infected cells, and other aspects of apoptosis are not triggered during early MVM infection. Instead, basally active caspase-3 is relocalized to the nuclei of infected cells. We propose that NE disruption involving caspases plays a role in (i) parvovirus entry into the nucleus and (ii) alteration of the compartmentalization of host proteins in a way that is favorable for the virus. PMID:21367902

Three-Dimensional Reconstruction of Nuclear Envelope Architecture Using Dual-Color Metal-Induced Energy Transfer Imaging.

PubMed

Chizhik, Anna M; Ruhlandt, Daja; Pfaff, Janine; Karedla, Narain; Chizhik, Alexey I; Gregor, Ingo; Kehlenbach, Ralph H; Enderlein, Jörg

2017-12-26

The nuclear envelope, comprising the inner and the outer nuclear membrane, separates the nucleus from the cytoplasm and plays a key role in cellular functions. Nuclear pore complexes (NPCs), which are embedded in the nuclear envelope, control transport of macromolecules between the two compartments. Here, using dual-color metal-induced energy transfer (MIET), we determine the axial distance between Lap2β and Nup358 as markers for the inner nuclear membrane and the cytoplasmic side of the NPC, respectively. Using MIET imaging, we reconstruct the 3D profile of the nuclear envelope over the whole basal area, with an axial resolution of a few nanometers. This result demonstrates that optical microscopy can achieve nanometer axial resolution in biological samples and without recourse to complex interferometric approaches.

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

PubMed

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.

Breaching the nuclear envelope in development and disease

PubMed Central

Hatch, Emily

2014-01-01

In eukaryotic cells the nuclear genome is enclosed by the nuclear envelope (NE). In metazoans, the NE breaks down in mitosis and it has been assumed that the physical barrier separating nucleoplasm and cytoplasm remains intact during the rest of the cell cycle and cell differentiation. However, recent studies suggest that nonmitotic NE remodeling plays a critical role in development, virus infection, laminopathies, and cancer. Although the mechanisms underlying these NE restructuring events are currently being defined, one common theme is activation of protein kinase C family members in the interphase nucleus to disrupt the nuclear lamina, demonstrating the importance of the lamina in maintaining nuclear integrity. PMID:24751535

Dynamic Assembly of Brambleberry Mediates Nuclear Envelope Fusion during Early Development

PubMed Central

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

2012-01-01

Summary To accommodate the large cells following zygote formation, early blastomeres employ modified cell divisions. Karyomeres are one such modification, a mitotic intermediate wherein individual chromatin masses are surrounded by nuclear envelope, which 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. brambleberry is a previously unannotated gene homologous to Kar5p, which participates in nuclear fusion in yeast. We demonstrate that Brambleberry is required for pronuclear fusion following fertilization in zebrafish. As karyomeres form, Brambleberry localizes to the nuclear envelope with prominent puncta evident near karyomere-karyomere interfaces corresponding to membrane fusion sites. Our studies identify the first factor acting in karyomere fusion and suggest that specialized proteins are necessary for proper nuclear division in large dividing blastomeres. PMID:22863006

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

PubMed Central

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. PMID:25932910

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.

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

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.

Old foes, new understandings: nuclear entry of small non-enveloped DNA viruses.

PubMed

Fay, Nikta; Panté, Nelly

2015-06-01

The nuclear import of viral genomes is an important step of the infectious cycle for viruses that replicate in the nucleus of their host cells. Although most viruses use the cellular nuclear import machinery or some components of this machinery, others have developed sophisticated ways to reach the nucleus. Some of these have been known for some time; however, recent studies have changed our understanding of how some non-enveloped DNA viruses access the nucleus. For example, parvoviruses enter the nucleus through small disruptions of the nuclear membranes and nuclear lamina, and adenovirus tugs at the nuclear pore complex, using kinesin-1, to disassemble their capsids and deliver viral proteins and genomes into the nucleus. Here we review recent findings of the nuclear import strategies of three small non-enveloped DNA viruses, including adenovirus, parvovirus, and the polyomavirus simian virus 40. Copyright © 2015 Elsevier B.V. All rights reserved.

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

2012-08-03

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. Copyright © 2012 Elsevier Inc. All rights reserved.

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

The nuclear envelope as an integrator of nuclear and cytoplasmic architecture.

PubMed

Crisp, Melissa; Burke, Brian

2008-06-18

Initially perceived as little more than a container for the genome, our view of the nuclear envelope (NE) and its role in defining global nuclear architecture has evolved significantly in recent years. The recognition that certain human diseases arise from defects in NE components has provided new insight into its structural and regulatory functions. In particular, NE defects associated with striated muscle disease have been shown to cause structural perturbations not just of the nucleus itself but also of the cytoplasm. It is now becoming increasingly apparent that these two compartments display co-dependent mechanical properties. The identification of cytoskeletal binding complexes that localize to the NE now reveals a molecular framework that can seamlessly integrate nuclear and cytoplasmic architecture.

STEF/TIAM2-mediated Rac1 activity at the nuclear envelope regulates the perinuclear actin cap.

PubMed

Woroniuk, Anna; Porter, Andrew; White, Gavin; Newman, Daniel T; Diamantopoulou, Zoi; Waring, Thomas; Rooney, Claire; Strathdee, Douglas; Marston, Daniel J; Hahn, Klaus M; Sansom, Owen J; Zech, Tobias; Malliri, Angeliki

2018-05-29

The perinuclear actin cap is an important cytoskeletal structure that regulates nuclear morphology and re-orientation during front-rear polarisation. The mechanisms regulating the actin cap are currently poorly understood. Here, we demonstrate that STEF/TIAM2, a Rac1 selective guanine nucleotide exchange factor, localises at the nuclear envelope, co-localising with the key perinuclear proteins Nesprin-2G and Non-muscle myosin IIB (NMMIIB), where it regulates perinuclear Rac1 activity. We show that STEF depletion reduces apical perinuclear actin cables (a phenotype rescued by targeting active Rac1 to the nuclear envelope), increases nuclear height and impairs nuclear re-orientation. STEF down-regulation also reduces perinuclear pMLC and decreases myosin-generated tension at the nuclear envelope, suggesting that STEF-mediated Rac1 activity regulates NMMIIB activity to promote stabilisation of the perinuclear actin cap. Finally, STEF depletion decreases nuclear stiffness and reduces expression of TAZ-regulated genes, indicating an alteration in mechanosensing pathways as a consequence of disruption of the actin cap.

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. Copyright © 2015 Elsevier Inc. All rights reserved.

Actomyosin drives cancer cell nuclear dysmorphia and threatens genome stability.

PubMed

Takaki, Tohru; Montagner, Marco; Serres, Murielle P; Le Berre, Maël; Russell, Matt; Collinson, Lucy; Szuhai, Karoly; Howell, Michael; Boulton, Simon J; Sahai, Erik; Petronczki, Mark

2017-07-24

Altered nuclear shape is a defining feature of cancer cells. The mechanisms underlying nuclear dysmorphia in cancer remain poorly understood. Here we identify PPP1R12A and PPP1CB, two subunits of the myosin phosphatase complex that antagonizes actomyosin contractility, as proteins safeguarding nuclear integrity. Loss of PPP1R12A or PPP1CB causes nuclear fragmentation, nuclear envelope rupture, nuclear compartment breakdown and genome instability. Pharmacological or genetic inhibition of actomyosin contractility restores nuclear architecture and genome integrity in cells lacking PPP1R12A or PPP1CB. We detect actin filaments at nuclear envelope rupture sites and define the Rho-ROCK pathway as the driver of nuclear damage. Lamin A protects nuclei from the impact of actomyosin activity. Blocking contractility increases nuclear circularity in cultured cancer cells and suppresses deformations of xenograft nuclei in vivo. We conclude that actomyosin contractility is a major determinant of nuclear shape and that unrestrained contractility causes nuclear dysmorphia, nuclear envelope rupture and genome instability.

Actomyosin drives cancer cell nuclear dysmorphia and threatens genome stability

PubMed Central

Takaki, Tohru; Montagner, Marco; Serres, Murielle P.; Le Berre, Maël; Russell, Matt; Collinson, Lucy; Szuhai, Karoly; Howell, Michael; Boulton, Simon J.; Sahai, Erik; Petronczki, Mark

2017-01-01

Altered nuclear shape is a defining feature of cancer cells. The mechanisms underlying nuclear dysmorphia in cancer remain poorly understood. Here we identify PPP1R12A and PPP1CB, two subunits of the myosin phosphatase complex that antagonizes actomyosin contractility, as proteins safeguarding nuclear integrity. Loss of PPP1R12A or PPP1CB causes nuclear fragmentation, nuclear envelope rupture, nuclear compartment breakdown and genome instability. Pharmacological or genetic inhibition of actomyosin contractility restores nuclear architecture and genome integrity in cells lacking PPP1R12A or PPP1CB. We detect actin filaments at nuclear envelope rupture sites and define the Rho-ROCK pathway as the driver of nuclear damage. Lamin A protects nuclei from the impact of actomyosin activity. Blocking contractility increases nuclear circularity in cultured cancer cells and suppresses deformations of xenograft nuclei in vivo. We conclude that actomyosin contractility is a major determinant of nuclear shape and that unrestrained contractility causes nuclear dysmorphia, nuclear envelope rupture and genome instability. PMID:28737169

The Role of the Nuclear Envelope Protein MAN1 in Mesenchymal Stem Cell Differentiation.

PubMed

Bermeo, Sandra; Al-Saedi, Ahmed; Kassem, Moustapha; Vidal, Christopher; Duque, Gustavo

2017-12-01

Mutations in MAN1, a protein of the nuclear envelope, cause bone phenotypes characterized by hyperostosis. The mechanism of this pro-osteogenic phenotype remains unknown. We increased and decreased MAN1 expression in mesenchymal stem cells (MSC) upon which standard osteogenic and adipogenic differentiation were performed. MAN1 knockdown increased osteogenesis and mineralization. In contrast, osteogenesis remained stable upon MAN1 overexpression. Regarding a mechanism, we found that low levels of MAN1 facilitated the nuclear accumulation of regulatory smads and smads-related complexes, with a concurrently high expression of nuclear β-Catenin. In addition, we found adipogenesis to be decreased in both conditions, although predominantly affected by MAN1 overexpression. Finally, lamin A, a protein of the nuclear envelope that regulates MSC differentiation, was unaffected by changes in MAN1. In conclusion, our studies demonstrated that lower levels of MAN1 in differentiating MSC are associated with higher osteogenesis and lower adipogenesis. High levels of MAN1 only affected adipogenesis. These effects could have an important role in the understanding of the role of the proteins of the nuclear envelope in bone formation. J. Cell. Biochem. 118: 4425-4435, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

LEM2 recruits CHMP7 for ESCRT-mediated nuclear envelope closure in fission yeast and human cells

PubMed Central

Gu, Mingyu; LaJoie, Dollie; Chen, Opal S.; von Appen, Alexander; Ladinsky, Mark S.; Redd, Michael J.; Nikolova, Linda; Bjorkman, Pamela J.; Sundquist, Wesley I.; Ullman, Katharine S.; Frost, Adam

2017-01-01

Endosomal sorting complexes required for transport III (ESCRT-III) proteins have been implicated in sealing the nuclear envelope in mammals, spindle pole body dynamics in fission yeast, and surveillance of defective nuclear pore complexes in budding yeast. Here, we report that Lem2p (LEM2), a member of the LEM (Lap2-Emerin-Man1) family of inner nuclear membrane proteins, and the ESCRT-II/ESCRT-III hybrid protein Cmp7p (CHMP7), work together to recruit additional ESCRT-III proteins to holes in the nuclear membrane. In Schizosaccharomyces pombe, deletion of the ATPase vps4 leads to severe defects in nuclear morphology and integrity. These phenotypes are suppressed by loss-of-function mutations that arise spontaneously in lem2 or cmp7, implying that these proteins may function upstream in the same pathway. Building on these genetic interactions, we explored the role of LEM2 during nuclear envelope reformation in human cells. We found that CHMP7 and LEM2 enrich at the same region of the chromatin disk periphery during this window of cell division and that CHMP7 can bind directly to the C-terminal domain of LEM2 in vitro. We further found that, during nuclear envelope formation, recruitment of the ESCRT factors CHMP7, CHMP2A, and IST1/CHMP8 all depend on LEM2 in human cells. We conclude that Lem2p/LEM2 is a conserved nuclear site-specific adaptor that recruits Cmp7p/CHMP7 and downstream ESCRT factors to the nuclear envelope. PMID:28242692

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

PubMed

Li, Ping; Noegel, Angelika A

2015-11-16

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 l I: nker of N: ucleoskeleton and C: ytoskeleton (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. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Regulation of nuclear envelope dynamics via APC/C is necessary for the progression of semi-open mitosis in Schizosaccharomyces japonicus.

PubMed

Aoki, Keita; Shiwa, Yuh; Takada, Hiraku; Yoshikawa, Hirofumi; Niki, Hironori

2013-09-01

Three types of mitosis, which are open, closed or semi-open mitosis, function in eukaryotic cells, respectively. The open mitosis involves breakage of the nuclear envelope before nuclear division, whereas the closed mitosis proceeds with an intact nuclear envelope. To understand the mechanism and significance of three types of mitotic division in eukaryotes, we investigated the process of semi-open mitosis, in which the nuclear envelope is only partially broken, in the fission yeast Schizosaccharomyces japonicus. In anaphase-promoting complex/cyclosome (APC/C) mutants of Sz. japonicus, the nuclear envelope remained relatively intact during anaphase, resulting in impaired semi-open mitosis. As a suppressor of apc2 mutant, a mutation of Oar2, which was a 3-oxoacyl-[acyl carrier protein] reductase, was obtained. The level of the Oar2, which had two destruction-box motifs recognized by APC/C, was increased in APC/C mutants. Furthermore, the defective semi-open mitosis observed in an apc2 mutant was restored by mutated oar2+. Based on these findings, we propose that APC/C regulates the dynamics of the nuclear envelope through degradation of Oar2 dependent on APC/C during the metaphase-to-anaphase transition of semi-open mitosis in Sz. japonicus. © 2013 The Authors Genes to Cells © 2013 by the Molecular Biology Society of Japan and Wiley Publishing Asia Pty Ltd.

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.

Nup155 regulates nuclear envelope and nuclear pore complex formation in nematodes and vertebrates

PubMed Central

Franz, Cerstin; Askjaer, Peter; Antonin, Wolfram; Iglesias, Carmen López; Haselmann, Uta; Schelder, Malgorzata; de Marco, Ario; Wilm, Matthias; Antony, Claude; Mattaj, Iain W

2005-01-01

Nuclear envelope (NE) formation during cell division in multicellular organisms is a central yet poorly understood biological process. We report that the conserved nucleoporin Nup155 has an essential function in NE formation in Caenorhabditis elegans embryos and in Xenopus laevis egg extracts. In vivo depletion of Nup155 led to failure of nuclear lamina formation and defects in chromosome segregation at anaphase. Nup155 depletion inhibited accumulation of nucleoporins at the nuclear periphery, including those recruited to chromatin early in NE formation. Electron microscopy analysis revealed that Nup155 is also required for the formation of a continuous nuclear membrane in vivo and in vitro. Time-course experiments indicated that Nup155 is recruited to chromatin at the time of NE sealing, suggesting that nuclear pore complex assembly has to progress to a relatively late stage before NE membrane assembly occurs. PMID:16193066

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. Copyright © 2015 Rogers and Rose.

Kar5p Is Required for Multiple Functions in Both Inner and Outer Nuclear Envelope Fusion in Saccharomyces cerevisiae

PubMed Central

Rogers, Jason V.; Rose, Mark D.

2014-01-01

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. PMID:25467943

NUCLEAR ENVELOPE-ASSOCIATED RESUMPTION OF RNA SYNTHESIS IN LATE MITOSIS OF HELA CELLS

PubMed Central

Simmons, T.; Heywood, P.; Hodge, L.

1973-01-01

The restitution of RNA synthesis in cultures progressing from metaphase into interphase (G1) has been investigated in synchronized HeLa S3 cells by using inhibitors of macro-molecular synthesis and the technique of electron microscope autoradiography. The rate of incorporation of radioactive uridine into RNA approached interphase levels in the absence of renewed protein synthesis. In contrast, maintenance of this rate in G1 was dependent upon renewed protein synthesis. Restoration of synthesis of heterogeneous nuclear RNA occurred under conditions that inhibited production of ribosomal precursor RNA. In autoradiographs of individual cells exposed to radioactive uridine, silver grains were first detected after nuclear envelope reformation at the periphery of the chromosome mass but before chromosomal decondensation. These data are consistent with the following interpretation. Multiple RNA polymerase activities persist through mitosis and are involved in the initiation of RNA synthesis in early telophase at sites on the nuclear envelope. PMID:4752403

Nuclear envelope rupture: little holes, big openings.

PubMed

Hatch, Emily M

2018-06-01

The nuclear envelope (NE), which is a critical barrier between the DNA and the cytosol, is capable of extensive dynamic membrane remodeling events in interphase. One of these events, interphase NE rupture and repair, can occur in both normal and disease states and results in the loss of nucleus compartmentalization. NE rupture is not lethal, but new research indicates that it could have broad impacts on genome stability and activate innate immune responses. These observations suggest a new model for how changes in NE structure could be pathogenic in cancer, laminopathies, and autoinflammatory syndromes, and redefine the functions of nucleus compartmentalization. Copyright © 2018 Elsevier Ltd. All rights reserved.

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

Improving nuclear envelope dynamics by EBV BFRF1 facilitates intranuclear component clearance through autophagy.

PubMed

Liu, Guan-Ting; Kung, Hsiu-Ni; Chen, Chung-Kuan; Huang, Cheng; Wang, Yung-Li; Yu, Cheng-Pu; Lee, Chung-Pei

2018-02-26

Although a vesicular nucleocytoplasmic transport system is believed to exist in eukaryotic cells, the features of this pathway are mostly unknown. Here, we report that the BFRF1 protein of the Epstein-Barr virus improves vesicular transport of nuclear envelope (NE) to facilitate the translocation and clearance of nuclear components. BFRF1 expression induces vesicles that selectively transport nuclear components to the cytoplasm. With the use of aggregation-prone proteins as tools, we found that aggregated nuclear proteins are dispersed when these BFRF1-induced vesicles are formed. BFRF1-containing vesicles engulf the NE-associated aggregates, exit through from the NE, and putatively fuse with autophagic vacuoles. Chemical treatment and genetic ablation of autophagy-related factors indicate that autophagosome formation and autophagy-linked FYVE protein-mediated autophagic proteolysis are involved in this selective clearance of nuclear proteins. Remarkably, vesicular transport, elicited by BFRF1, also attenuated nuclear aggregates accumulated in neuroblastoma cells. Accordingly, induction of NE-derived vesicles by BFRF1 facilitates nuclear protein translocation and clearance, suggesting that autophagy-coupled transport of nucleus-derived vesicles can be elicited for nuclear component catabolism in mammalian cells.-Liu, G.-T., Kung, H.-N., Chen, C.-K., Huang, C., Wang, Y.-L., Yu, C.-P., Lee, C.-P. Improving nuclear envelope dynamics by EBV BFRF1 facilitates intranuclear component clearance through autophagy.

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.

Data on the association of the nuclear envelope protein Sun1 with nucleoli.

PubMed

Moujaber, Ossama; Omran, Nawal; Kodiha, Mohamed; Pié, Brigitte; Cooper, Ellis; Presley, John F; Stochaj, Ursula

2017-08-01

SUN proteins participate in diverse cellular activities, many of which are connected to the nuclear envelope. Recently, the family member SUN1 has been linked to novel biological activities. These include the regulation of nucleoli, intranuclear compartments that assemble ribosomal subunits. We show that SUN1 associates with nucleoli in several mammalian epithelial cell lines. This nucleolar localization is not shared by all cell types, as SUN1 concentrates at the nuclear envelope in ganglionic neurons and non-neuronal satellite cells. Database analyses and Western blotting emphasize the complexity of SUN1 protein profiles in different mammalian cells. We constructed a STRING network which identifies SUN1-related proteins as part of a larger network that includes several nucleolar proteins. Taken together, the current data highlight the diversity of SUN1 proteins and emphasize the possible links between SUN1 and nucleoli.

Aspects of nuclear envelope dynamics in mitotic cells.

PubMed

Burke, Brian; Shanahan, Catherine; Salina, Davide; Crisp, Melissa

2005-01-01

Major features of the nuclear envelope (NE) are a pair of inner and outer nuclear membranes (INM, ONM) spanned by nuclear pore complexes. While the composition of the ONM resembles that of the endoplasmic reticulum, the INM contains a unique spectrum of proteins. Localization of INM proteins involves a mechanism of selective retention whereby integral proteins are immobilized and concentrated by virtue of interactions with nuclear components. In the case of emerin, INM localization involves interaction with A-type lamins. Interactions between membrane proteins may also play a significant role in INM localization. This conclusion stems from studies on nesprins, a family of membrane proteins that feature a large cytoplasmic domain, a single C-terminal membrane-spanning domain and a small lumenal domain. The nesprin membrane anchor and lumenal (KASH) domains are related to the Drosophila Klarsicht protein. Evidence is emerging that this KASH region interacts with other NE proteins and may influence their distributions. Overexpression of GFP-KASH causes loss of emerin and LAP2 from the NE. This is not due to global reorganization of the NE since LAP1 as well as lamins and NPCs remain unaffected. Our results suggest that interactions between NE membrane components are far more extensive and complex than current models suggest.

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

PubMed Central

Marshall, Wallace F.; Fung, Jennifer C.

2016-01-01

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 un-attached chromosomes, but that randomly-directed active forces applied to the telomeres speeds 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. PMID:27046097

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

Venture from the Interior-Herpesvirus pUL31 Escorts Capsids from Nucleoplasmic Replication Compartments to Sites of Primary Envelopment at the Inner Nuclear Membrane.

PubMed

Bailer, Susanne M.

2017-11-25

Herpesviral capsid assembly is initiated in the nucleoplasm of the infected cell. Size constraints require that newly formed viral nucleocapsids leave the nucleus by an evolutionarily conserved vescular transport mechanism called nuclear egress. Mature capsids released from the nucleoplasm are engaged in a membrane-mediated budding process, composed of primary envelopment at the inner nuclear membrane and de-envelopment at the outer nuclear membrane. Once in the cytoplasm, the capsids receive their secondary envelope for maturation into infectious virions. Two viral proteins conserved throughout the herpesvirus family, the integral membrane protein pUL34 and the phosphoprotein pUL31, form the nuclear egress complex required for capsid transport from the infected nucleus to the cytoplasm. Formation of the nuclear egress complex results in budding of membrane vesicles revealing its function as minimal virus-encoded membrane budding and scission machinery. The recent structural analysis unraveled details of the heterodimeric nuclear egress complex and the hexagonal coat it forms at the inside of budding vesicles to drive primary envelopment. With this review, I would like to present the capsid-escort-model where pUL31 associates with capsids in nucleoplasmic replication compartments for escort to sites of primary envelopment thereby coupling capsid maturation and nuclear egress.

Glimpsing over the event horizon: evolution of nuclear pores and envelope.

PubMed

Jékely, Gáspár

2005-02-01

The origin of eukaryotes from prokaryotic ancestors is one of the major evolutionary transitions in the history of life. The nucleus, a membrane bound compartment for confining the genome, is a central feature of eukaryotic cells and its origin also has to be a central feature of any workable theory that ventures to explain eukaryotic origins. Recent bioinformatic analyses of components of the nuclear pore complex (NPC), the nuclear envelope (NE), and the nuclear transport systems revealed exciting evolutionary connections (e.g., between NPC and coated vesicles) and provided a useful record of the phyletic distribution and history of NPC and NE components. These analyses allow us to refine theories on the origin and evolution of the nucleus, and consequently, of the eukaryotic cell.

Control of nuclear β-dystroglycan content is crucial for the maintenance of nuclear envelope integrity and function.

PubMed

Vélez-Aguilera, Griselda; de Dios Gómez-López, Juan; Jiménez-Gutiérrez, Guadalupe E; Vásquez-Limeta, Alejandra; Laredo-Cisneros, Marco S; Gómez, Pablo; Winder, Steve J; Cisneros, Bulmaro

2018-02-01

β-Dystroglycan (β-DG) is a plasma membrane protein that has ability to target to the nuclear envelope (NE) to maintain nuclear architecture. Nevertheless, mechanisms controlling β-DG nuclear localization and the physiological consequences of a failure of trafficking are largely unknown. We show that β-DG has a nuclear export pathway in myoblasts that depends on the recognition of a nuclear export signal located in its transmembrane domain, by CRM1. Remarkably, NES mutations forced β-DG nuclear accumulation resulting in mislocalization and decreased levels of emerin and lamin B1 and disruption of various nuclear processes in which emerin (centrosome-nucleus linkage and β-catenin transcriptional activity) and lamin B1 (cell cycle progression and nucleoli structure) are critically involved. In addition to nuclear export, the lifespan of nuclear β-DG is restricted by its nuclear proteasomal degradation. Collectively our data show that control of nuclear β-DG content by the combination of CRM1 nuclear export and nuclear proteasome pathways is physiologically relevant to preserve proper NE structure and activity. Copyright © 2017 Elsevier B.V. All rights reserved.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ostlund, Cecilia; Department of Pathology and Cell Biology, College of Physicians and Surgeons, Columbia University, New York, NY 10032; Guan, Tinglu

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 withmore » 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.« less

Induction of polyploidy by nuclear fusion mechanism upon decreased expression of the nuclear envelope protein LAP2β in the human osteosarcoma cell line U2OS.

PubMed

Ben-Shoshan, Shirley Oren; Simon, Amos J; Jacob-Hirsch, Jasmine; Shaklai, Sigal; Paz-Yaacov, Nurit; Amariglio, Ninette; Rechavi, Gideon; Trakhtenbrot, Luba

2014-01-28

Polyploidy has been recognized for many years as an important hallmark of cancer cells. Polyploid cells can arise through cell fusion, endoreplication and abortive cell cycle. The inner nuclear membrane protein LAP2β plays key roles in nuclear envelope breakdown and reassembly during mitosis, initiation of replication and transcriptional repression. Here we studied the function of LAP2β in the maintenance of cell ploidy state, a role which has not yet been assigned to this protein. By knocking down the expression of LAP2β, using both viral and non-viral RNAi approaches in osteosarcoma derived U2OS cells, we detected enlarged nuclear size, nearly doubling of DNA content and chromosomal duplications, as analyzed by fluorescent in situ hybridization and spectral karyotyping methodologies. Spectral karyotyping analyses revealed that near-hexaploid karyotypes of LAP2β knocked down cells consisted of not only seven duplicated chromosomal markers, as could be anticipated by genome duplication mechanism, but also of four single chromosomal markers. Furthermore, spectral karyotyping analysis revealed that both of two near-triploid U2OS sub-clones contained the seven markers that were duplicated in LAP2β knocked down cells, whereas the four single chromosomal markers were detected only in one of them. Gene expression profiling of LAP2β knocked down cells revealed that up to a third of the genes exhibiting significant changes in their expression are involved in cancer progression. Our results suggest that nuclear fusion mechanism underlies the polyploidization induction upon LAP2β reduced expression. Our study implies on a novel role of LAP2β in the maintenance of cell ploidy status. LAP2β depleted U2OS cells can serve as a model to investigate polyploidy and aneuploidy formation by nuclear fusion mechanism and its involvement in cancerogenesis.

Abnormal nuclear envelope in the cerebellar Purkinje cells and impaired motor learning in DYT11 myoclonus-dystonia mouse models

PubMed Central

Yokoi, Fumiaki; Dang, Mai T.; Yang, Guang; Li, JinDong; Doroodchi, Atbin; Zhou, Tong; Li, Yuqing

2011-01-01

Myoclonus-dystonia (M-D) is a movement disorder characterized by myoclonic jerks with dystonia. DYT11 M-D is caused by mutations in SGCE which codes for ε-sarcoglycan. SGCE is maternally imprinted and paternally expressed. Abnormal nuclear envelope has been reported in mouse models of DYT1 generalized torsion dystonia. However, it is not known whether similar alterations occur in DYT11 M-D. We developed a mouse model of DYT11 M-D using paternally-inherited Sgce heterozygous knockout (Sgce KO) mice and reported that they had myoclonus and motor coordination and learning deficits in the beam-walking test. However, the specific brain regions that contribute to these phenotypes have not been identified. Since ε-sarcoglycan is highly expressed in the cerebellar Purkinje cells, here we examined the nuclear envelope in these cells using a transmission electron microscope and found that they are abnormal in Sgce KO mice. Our results put DYT11 M-D in a growing family of nuclear envelopathies. To analyze the effect of loss of ε-sarcoglycan function in the cerebellar Purkinje cells, we produced paternally-inherited cerebellar Purkinje cell-specific Sgce conditional knockout (Sgce pKO) mice. Sgce pKO mice showed motor learning deficits, while they did not show abnormal nuclear envelope in the cerebellar Purkinje cells, robust motor deficits, or myoclonus. The results suggest that ε-sarcoglycan in the cerebellar Purkinje cells contributes to the motor learning, while loss of ε-sarcoglycan in other brain regions may contribute to nuclear envelope abnormality, myoclonus and motor coordination deficits. PMID:22040906

Outer nuclear membrane protein Kuduk modulates the LINC complex and nuclear envelope architecture

PubMed Central

Ding, Zhao-Ying; Huang, Yu-Cheng; Lee, Myong-Chol; Tseng, Min-Jen; Chi, Ya-Hui

2017-01-01

Linker of nucleoskeleton and cytoskeleton (LINC) complexes spanning the nuclear envelope (NE) contribute to nucleocytoskeletal force transduction. A few NE proteins have been found to regulate the LINC complex. In this study, we identify one, Kuduk (Kud), which can reside at the outer nuclear membrane and is required for the development of Drosophila melanogaster ovarian follicles and NE morphology of myonuclei. Kud associates with LINC complex components in an evolutionarily conserved manner. Loss of Kud increases the level but impairs functioning of the LINC complex. Overexpression of Kud suppresses NE targeting of cytoskeleton-free LINC complexes. Thus, Kud acts as a quality control mechanism for LINC-mediated nucleocytoskeletal connections. Genetic data indicate that Kud also functions independently of the LINC complex. Overexpression of the human orthologue TMEM258 in Drosophila proved functional conservation. These findings expand our understanding of the regulation of LINC complexes and NE architecture. PMID:28716842

Dynamic nuclear envelope phenotype in rats overexpressing mutated human torsinA protein.

PubMed

Yu-Taeger, Libo; Gaiser, Viktoria; Lotzer, Larissa; Roenisch, Tina; Fabry, Benedikt Timo; Stricker-Shaver, Janice; Casadei, Nicolas; Walter, Michael; Schaller, Martin; Riess, Olaf; Nguyen, Huu Phuc; Ott, Thomas; Grundmann-Hauser, Kathrin

2018-05-08

A three-base-pair deletion in the human TOR1A gene is causative for the most common form of primary dystonia, the early-onset dystonia type 1 (DYT1 dystonia). The pathophysiological consequences of this mutation are still unknown.To study the pathology of the mutant torsinA (TOR1A) protein, we have generated a transgenic rat line that overexpresses the human mutant protein under the control of the human TOR1A promoter. This new animal model was phenotyped with several approaches, including behavioral tests and neuropathological analyses. A motor phenotype and cellular and ultrastructural key features of torsinA pathology were found in this new transgenic rat line supporting that it can be used as a model system for investigating the disease development. Analyses of mutant TOR1A protein expression in various brain regions also showed a dynamic expression pattern and a reversible nuclear envelope pathology. These findings suggest the differential vulnerabilities of distinct neuronal subpopulations. Furthermore the reversibility of the nuclear envelope pathology might be a therapeutic target to treat the disease. © 2018. Published by The Company of Biologists Ltd.

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

PubMed

Popken, Jens; Schmid, Volker J; Strauss, Axel; Guengoer, Tuna; Wolf, Eckhard; Zakhartchenko, Valeri

2016-04-22

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

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

Abnormal nuclear envelope in the cerebellar Purkinje cells and impaired motor learning in DYT11 myoclonus-dystonia mouse models.

PubMed

Yokoi, Fumiaki; Dang, Mai T; Yang, Guang; Li, Jindong; Doroodchi, Atbin; Zhou, Tong; Li, Yuqing

2012-02-01

Myoclonus-dystonia (M-D) is a movement disorder characterized by myoclonic jerks with dystonia. DYT11 M-D is caused by mutations in SGCE which codes for ɛ-sarcoglycan. SGCE is maternally imprinted and paternally expressed. Abnormal nuclear envelope has been reported in mouse models of DYT1 generalized torsion dystonia. However, it is not known whether similar alterations occur in DYT11 M-D. We developed a mouse model of DYT11 M-D using paternally inherited Sgce heterozygous knockout (Sgce KO) mice and reported that they had myoclonus and motor coordination and learning deficits in the beam-walking test. However, the specific brain regions that contribute to these phenotypes have not been identified. Since ɛ-sarcoglycan is highly expressed in the cerebellar Purkinje cells, here we examined the nuclear envelope in these cells using a transmission electron microscope and found that they are abnormal in Sgce KO mice. Our results put DYT11 M-D in a growing family of nuclear envelopathies. To analyze the effect of loss of ɛ-sarcoglycan function in the cerebellar Purkinje cells, we produced paternally inherited cerebellar Purkinje cell-specific Sgce conditional knockout (Sgce pKO) mice. Sgce pKO mice showed motor learning deficits, while they did not show abnormal nuclear envelope in the cerebellar Purkinje cells, robust motor deficits, or myoclonus. The results suggest that ɛ-sarcoglycan in the cerebellar Purkinje cells contributes to the motor learning, while loss of ɛ-sarcoglycan in other brain regions may contribute to nuclear envelope abnormality, myoclonus and motor coordination deficits. Copyright © 2011 Elsevier B.V. All rights reserved.

Nup53 Is Required for Nuclear Envelope and Nuclear Pore Complex Assembly

PubMed Central

Hawryluk-Gara, Lisa A.; Platani, Melpomeni; Santarella, Rachel

2008-01-01

Transport across the nuclear envelope (NE) is mediated by nuclear pore complexes (NPCs). These structures are composed of various subcomplexes of proteins that are each present in multiple copies and together establish the eightfold symmetry of the NPC. One evolutionarily conserved subcomplex of the NPC contains the nucleoporins Nup53 and Nup155. Using truncation analysis, we have defined regions of Nup53 that bind to neighboring nucleoporins as well as those domains that target Nup53 to the NPC in vivo. Using this information, we investigated the role of Nup53 in NE and NPC assembly using Xenopus egg extracts. We show that both events require Nup53. Importantly, the analysis of Nup53 fragments revealed that the assembly activity of Nup53 depleted extracts could be reconstituted using a region of Nup53 that binds specifically to its interacting partner Nup155. On the basis of these results, we propose that the formation of a Nup53–Nup155 complex plays a critical role in the processes of NPC and NE assembly. PMID:18256286

Analysis of RNA-Seq datasets reveals enrichment of tissue-specific splice variants for nuclear envelope proteins.

PubMed

Capitanchik, Charlotte; Dixon, Charles; Swanson, Selene K; Florens, Laurence; Kerr, Alastair R W; Schirmer, Eric C

2018-06-18

Nuclear envelopathies/laminopathies yield tissue-specific pathologies, yet arise from mutation of ubiquitously-expressed genes. One possible explanation of this tissue specificity is that tissue-specific partners become disrupted from larger complexes, but a little investigated alternate hypothesis is that the mutated proteins themselves have tissue-specific splice variants. Here, we analyze RNA-Seq datasets to identify muscle-specific splice variants of nuclear envelope genes that could be relevant to the study of laminopathies, particularly muscular dystrophies, that are not currently annotated in sequence databases. Notably, we found novel isoforms or tissue-specificity of isoforms for: Lap2, linked to cardiomyopathy; Nesprin 2, linked to Emery-Dreifuss muscular dystrophy and Lmo7, a regulator of the emerin gene that is linked to Emery-Dreifuss muscular dystrophy. Interestingly, the muscle-specific exon in Lmo7 is rich in serine phosphorylation motifs, suggesting an important regulatory function. Evidence for muscle-specific splice variants in non-nuclear envelope proteins linked to other muscular dystrophies was also found. Tissue-specific variants were also indicated for several nucleoporins including Nup54, Nup133, Nup153 and Nup358/RanBP2. We confirmed expression of novel Lmo7 and RanBP2 variants with RT-PCR and found that specific knockdown of the Lmo7 variant caused a reduction in myogenic index during mouse C2C12 myogenesis. Global analysis revealed an enrichment of tissue-specific splice variants for nuclear envelope proteins in general compared to the rest of the genome, suggesting that splice variants contribute to regulating its tissue-specific functions.

Overexpression of the lamina proteins Lamin and Kugelkern induces specific ultrastructural alterations in the morphology of the nuclear envelope of intestinal stem cells and enterocytes.

PubMed

Petrovsky, Roman; Krohne, Georg; Großhans, Jörg

2018-03-01

The nuclear envelope has a stereotypic morphology consisting of a flat double layer of the inner and outer nuclear membrane, with interspersed nuclear pores. Underlying and tightly linked to the inner nuclear membrane is the nuclear lamina, a proteinous layer of intermediate filament proteins and associated proteins. Physiological, experimental or pathological alterations in the constitution of the lamina lead to changes in nuclear morphology, such as blebs and lobulations. It has so far remained unclear whether the morphological changes depend on the differentiation state and the specific lamina protein. Here we analysed the ultrastructural morphology of the nuclear envelope in intestinal stem cells and differentiated enterocytes in adult Drosophila flies, in which the proteins Lam, Kugelkern or a farnesylated variant of LamC were overexpressed. Surprisingly, we detected distinct morphological features specific for the respective protein. Lam induced envelopes with multiple layers of membrane and lamina, surrounding the whole nucleus whereas farnesylated LamC induced the formation of a thick fibrillary lamina. In contrast, Kugelkern induced single-layered and double-layered intranuclear membrane structures, which are likely be derived from infoldings of the inner nuclear membrane or of the double layer of the envelope. Copyright © 2018 Elsevier GmbH. All rights reserved.

Differential detection of nuclear envelope autoantibodies in primary biliary cirrhosis using routine and alternative methods

PubMed Central

2010-01-01

Background Detection of autoantibodies giving nuclear rim pattern by immunofluorescence (anti-nuclear envelope antibodies - ANEA) in sera from patients with primary biliary cirrhosis (PBC) is a useful tool for the diagnosis and prognosis of the disease. Differences in the prevalence of ANEA in PBC sera so far reported have been attributed to the methodology used for the detection as well as to ethnic/geographical variations. Therefore, we evaluated the prevalence of ANEA in sera of Greek patients with PBC by using methods widely used by clinical laboratories and a combination of techniques and materials. Methods We screened 103 sera by immunoblotting on nuclear envelopes and indirect immunofluorescence (IIF) using cells and purified nuclei. Reactivities against specific autoantigens were assessed using purified proteins, ELISA, immunoprecipitation and mass spectrometry. Results We found higher prevalence of ANEA when sera were assayed by IIF on purified nuclei or cultured cells (50%) compared to Hep2 commercially available slides (15%). Anti-gp210 antibodies were identified in 22.3% and 33% of sera using ELISA for the C-terminal of gp210 or both ELISA and immunoprecipitation, respectively. Immunoblotting on nuclear envelopes revealed that immunoreactivity for the 210 kDa zone is related to anti-gp210 antibodies (p < 0.0001). Moreover, we found that sera had antibodies for lamins A (6.8%), B (1%) and C (1%) and LBR (8.7%), whereas none at all had detectable anti-p62 antibodies. Conclusions The prevalence of ANEA or anti-gp210 antibodies is under-estimated in PBC sera which are analyzed by conventional commercially available IIF or ELISA, respectively. Therefore, new substrates for IIF and ELISA should be included by clinical laboratories in the analysis of ANEA in autoimmune sera. PMID:20205958

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

PubMed Central

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

ABSTRACT 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

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.

Mechanism of protein import across the chloroplast envelope.

PubMed

Chen, K; Chen, X; Schnell, D J

2000-01-01

The development and maintenance of chloroplasts relies on the contribution of protein subunits from both plastid and nuclear genomes. Most chloroplast proteins are encoded by nuclear genes and are post-translationally imported into the organelle across the double membrane of the chloroplast envelope. Protein import into the chloroplast consists of two essential elements: the specific recognition of the targeting signals (transit sequences) of cytoplasmic preproteins by receptors at the outer envelope membrane and the subsequent translocation of preproteins simultaneously across the double membrane of the envelope. These processes are mediated via the co-ordinate action of protein translocon complexes in the outer (Toc apparatus) and inner (Tic apparatus) envelope membranes.

Shape Transformation of the Nuclear Envelope during Closed Mitosis.

PubMed

Zhu, Qian; Zheng, Fan; Liu, Allen P; Qian, Jin; Fu, Chuanhai; Lin, Yuan

2016-11-15

The nuclear envelope (NE) in lower eukaryotes such as Schizosaccharomyces pombe undergoes large morphology changes during closed mitosis. However, which physical parameters are important in governing the shape evolution of the NE, and how defects in the dividing chromosomes/microtubules are reflected in those parameters, are fundamental questions that remain unresolved. In this study, we show that improper separation of chromosomes in genetically deficient cells leads to membrane tethering or asymmetric division in contrast to the formation of two equal-sized daughter nuclei in wild-type cells. We hypothesize that the poleward force is transmitted to the nuclear membrane through its physical contact with the separated sister chromatids at the two spindle poles. A theoretical model is developed to predict the morphology evolution of the NE where key factors such as the work done by the poleward force and bending and surface energies stored in the membrane have been taken into account. Interestingly, the predicted phase diagram, summarizing the dependence of nuclear shape on the size of the load transmission regions, and the pole-to-pole distance versus surface area relationship all quantitatively agree well with our experimental observations, suggesting that this model captures the essential physics involved in closed mitosis. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Pushing the envelope: microinjection of Minute virus of mice into Xenopus oocytes causes damage to the nuclear envelope.

PubMed

Cohen, Sarah; Panté, Nelly

2005-12-01

Parvoviruses are small DNA viruses that replicate in the nucleus of their host cells. It has been largely assumed that parvoviruses enter the nucleus through the nuclear pore complex (NPC). However, the details of this mechanism remain undefined. To study this problem, the parvovirus Minute virus of mice (MVM) was microinjected into the cytoplasm of Xenopus oocytes and a transmission electron microscope was used to visualize the effect of the virus on the host cell. It was found that MVM caused damage to the nuclear envelope (NE) in a time- and concentration-dependent manner. Damage was predominantly to the outer nuclear membrane and was often near the NPCs. However, microinjection experiments in which the NPCs were blocked showed that NE damage induced by MVM was independent of the NPC. To address the question of whether this effect of MVM is specific to the NE, purified organelles were incubated with MVM. Visualization by electron microscopy revealed that MVM did not affect all intracellular membranes. These data represent a novel form of virus-induced damage to host cell nuclear structure and suggest that MVM is imported into the nucleus using a unique mechanism that is independent of the NPC, and involves disruption of the NE and import through the resulting breaks.

Dynein pulling forces counteract lamin-mediated nuclear stability during nuclear envelope repair

PubMed Central

Penfield, Lauren; Wysolmerski, Brian; Mauro, Michael; Farhadifar, Reza; Martinez, Michael A.; Biggs, Ronald; Wu, Hai-Yin; Broberg, Curtis; Needleman, Daniel; Bahmanyar, Shirin

2018-01-01

Recent work done exclusively in tissue culture cells revealed that the nuclear envelope (NE) ruptures and repairs in interphase. The duration of NE ruptures depends on lamins; however, the underlying mechanisms and relevance to in vivo events are not known. Here, we use the Caenorhabditis elegans zygote to analyze lamin’s role in NE rupture and repair in vivo. Transient NE ruptures and subsequent NE collapse are induced by weaknesses in the nuclear lamina caused by expression of an engineered hypomorphic C. elegans lamin allele. Dynein-generated forces that position nuclei enhance the severity of transient NE ruptures and cause NE collapse. Reduction of dynein forces allows the weakened lamin network to restrict nucleo–cytoplasmic mixing and support stable NE recovery. Surprisingly, the high incidence of transient NE ruptures does not contribute to embryonic lethality, which is instead correlated with stochastic chromosome scattering resulting from premature NE collapse, suggesting that C. elegans tolerates transient losses of NE compartmentalization during early embryogenesis. In sum, we demonstrate that lamin counteracts dynein forces to promote stable NE repair and prevent catastrophic NE collapse, and thus provide the first mechanistic analysis of NE rupture and repair in an organismal context. PMID:29386297

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.

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

Built-up outer wall and roofing sections for double walled envelope homes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brodhead, B.

1980-01-01

A site built system that uses the inner envelope wall is described. Blocking and vertical nailers are attached to this wall and sheathed with foil faced drywall to create the envelope cavity. An outer layer of 3 1/2 in. of Expended Poly Styrene provides continuous solid insulation. The trusses are also sheathed in foil faced drywall and insulated with 5 1/2 in. of E.P.S. This effectively surrounds the building with a continuous vapor and infiltration barrier. Construction details as well as cost breakdowns are presented.

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

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. © 2016. Published by The Company of Biologists Ltd.

Yeast Nucleoporins Involved in Passive Nuclear Envelope Permeability

PubMed Central

Shulga, Nataliya; Mosammaparast, Nima; Wozniak, Richard; Goldfarb, David S.

2000-01-01

The vertebrate nuclear pore complex (NPC) harbors an ∼10-nm diameter diffusion channel that is large enough to admit 50-kD polypeptides. We have analyzed the permeability properties of the Saccharomyces cerevisiae nuclear envelope (NE) using import (NLS) and export (NES) signal-containing green fluorescent protein (GFP) reporters. Compared with wild-type, passive export rates of a classical karyopherin/importin (Kap) Kap60p/Kap95p-targeted NLS-GFP reporter (cNLS-GFP) were significantly faster in nup188-Δ and nup170-Δ cells. Similar results were obtained using two other NLS-GFP reporters, containing either the Kap104p-targeted Nab2p NLS (rgNLS) or the Kap121p-targeted Pho4p NLS (pNLS). Elevated levels of Hsp70 stimulated cNLS-GFP import, but had no effect on the import of rgNLS-GFP. Thus, the role of Hsp70 in NLS-directed import may be NLS- or targeting pathway-specific. Equilibrium sieving limits for the diffusion channel were assessed in vivo using NES-GFP reporters of 36–126 kD and were found to be greater than wild-type in nup188-Δ and nup170-Δ cells. We propose that Nup170p and Nup188p are involved in establishing the functional resting diameter of the NPC's central transport channel. PMID:10831607

A Novel Fission Yeast Gene, tht1 +, Is Required for the Fusion of Nuclear Envelopes during Karyogamy

PubMed Central

Tange, Yoshie; Horio, Tetsuya; Shimanuki, Mizuki; Ding, Da-Qiao; Hiraoka, Yasushi; Niwa, Osami

1998-01-01

We have isolated a fission yeast karyogamy mutant, tht1, in which nuclear congression and the association of two spindle pole bodies occurs but the subsequent fusion of nuclear envelopes is blocked. The tht1 mutation does not prevent meiosis, so cells execute meiosis with two unfused nuclei, leading to the production of aberrant asci. The tht1 + gene was cloned and sequenced. Predicted amino acid sequence has no significant homology to previously known proteins but strongly suggests that it is a type I membrane protein. The tht1 + gene is dispensable for vegetative growth and expressed only in conjugating cells. Tht1p is a glycoprotein susceptible to endoglycosilase H digestion. Site- directed mutagenesis showed that the N-glycosylation site, as well as the COOH-terminal region of Tht1p, is essential for its function. A protease protection assay indicated that the COOH terminus is cytoplasmic. Immunocytological analysis using a HA-tagged Tht1p suggested that the protein is localized in nuclear envelopes and in the ER during karyogamy and that its levels are reduced in cells containing fused nuclei. PMID:9442101

Ubiquitin-dependent Protein Degradation at the Yeast Endoplasmic Reticulum and Nuclear Envelope

PubMed Central

Zattas, Dimitrios; Hochstrasser, Mark

2014-01-01

The endoplasmic reticulum (ER) is the primary organelle in eukaryotic cells where membrane and secreted proteins are inserted into or across cell membranes. Its membrane bilayer and luminal compartments provide a favorable environment for the folding and assembly of thousands of newly synthesized proteins. However, protein folding is intrinsically error-prone, and various stress conditions can further increase levels of protein misfolding and damage, particularly in the ER, which can lead to cellular dysfunction and disease. The ubiquitin-proteasome system (UPS) is responsible for the selective destruction of a vast array of protein substrates, either for protein quality control or to allow rapid changes in the levels of specific regulatory proteins. In this review, we will focus on the components and mechanisms of ER-associated protein degradation (ERAD), an important branch of the UPS. ER membranes extend from subcortical regions of the cell to the nuclear envelope, with its continuous outer and inner membranes; the nuclear envelope is a specialized subdomain of the ER. ERAD presents additional challenges to the UPS beyond those faced with soluble substrates of the cytoplasm and nucleus. These include recognition of sugar modifications that occur in the ER, retrotranslocation of proteins across the membrane bilayer, and transfer of substrates from the ER extraction machinery to the proteasome. Here we review characteristics of ERAD substrate degradation signals (degrons), mechanisms underlying substrate recognition and processing by the ERAD machinery, and ideas on the still unresolved problem of how substrate proteins are moved across and extracted from the ER membrane. PMID:25231236

The fate and origin of the nuclear envelope during and after mitosis in Amoeba proteus. I. Synthesis and behavior of phospholipids of the nuclear envelope during the cell life cycle

PubMed Central

1975-01-01

The synthesis and behavior of Amoeba proteus nuclear envelope (NE) phospholipids were studied. Most NE phospholipid synthesis occurs during G2 and little during mitosis or S. (A. proteus has no G1 phase). Autoradiographic observations after implantation of [3-H] choline nuclei into unlabeled cells reveal little turnover of NE phospholipid during interphase but during mitosis all the label is dispersed through the cytoplasm. Beginning at telophase all the label is dispersed through the cytoplasm. Beginning at telophase all the NE phospholipid label returns to the daughter NEs. This observation, along with the finding that no NE phospholipid synthesis occurs during mitosis or S, indicates that no de novo NE phospholipid production is required for newly forming NEs. Similarlyemetine, at concentrations that inhibit 97 percent of protein synthesis, does not prevent the post mitotic formation of NEs, suggesting that previously manufactured proteins are used in making new NEs. If a nucleus containing labeled NE phospholipids is transplanted into an unlabeled nucleate cell and the cell is allowed to grow and divide, the resultant four nuclei are equally labeled. This finding supports, but does not prove (see next paragraph), the conclusion that there probably is no continuity of the A. proteus NE during mitosis. When a phospholipid-labeled nucleus is implanted into a cell in mitosis, the grafted nucleus is not induced to enter mitosis. There is, however, a marked increase in the turnover of that nucleus's NE phospholipids with no apparent breakdown of the NE; this indicated that the mitotic cytoplasm possesses a factor that stimulates NE phospholipid exchange with the cytoplasm. That enhanced turnover is not accompanied by visible structural alteration makes less certain the earlier conclusion that no NE continuity exists during mitosis. Perhaps the most important finding in this study is that there are present, at restricted times in the cell cycle, factors capable of

Seipin is involved in the regulation of phosphatidic acid metabolism at a subdomain of the nuclear envelope in yeast.

PubMed

Wolinski, Heimo; Hofbauer, Harald F; Hellauer, Klara; Cristobal-Sarramian, Alvaro; Kolb, Dagmar; Radulovic, Maja; Knittelfelder, Oskar L; Rechberger, Gerald N; Kohlwein, Sepp D

2015-11-01

Yeast Fld1 and Ldb16 resemble mammalian seipin, implicated in neutral lipid storage. Both proteins form a complex at the endoplasmic reticulum-lipid droplet (LD) interface. Malfunction of this complex either leads to LD clustering or to the generation of supersized LD (SLD) in close vicinity to the nuclear envelope, in response to altered phospholipid (PL) composition. We show that similar to mutants lacking Fld1, deletion of LDB16 leads to abnormal proliferation of a subdomain of the nuclear envelope, which is tightly associated with clustered LD. The human lipin-1 ortholog, the PAH1 encoded phosphatidic acid (PA) phosphatase, and its activator Nem1 are highly enriched at this site. The specific accumulation of PA-binding marker proteins indicates a local enrichment of PA in the fld1 and ldb16 mutants. Furthermore, we demonstrate that clustered LD in fld1 or ldb16 mutants are transformed to SLD if phosphatidylcholine synthesis is compromised by additional deletion of the phosphatidylethanolamine methyltransferase, Cho2. Notably, treatment of wild-type cells with oleate induced a similar LD clustering and nuclear membrane proliferation phenotype as observed in fld1 and ldb16 mutants. These data suggest that the Fld1-Ldb16 complex affects PA homeostasis at an LD-forming subdomain of the nuclear envelope. Lack of Fld1-Ldb16 leads to locally elevated PA levels that induce an abnormal proliferation of nER membrane structures and the clustering of associated LD. We suggest that the formation of SLD is a consequence of locally altered PL metabolism at this site. Copyright © 2015. Published by Elsevier B.V.

Bidirectional Increase in Permeability of Nuclear Envelope upon Poliovirus Infection and Accompanying Alterations of Nuclear Pores

PubMed Central

Belov, George A.; Lidsky, Peter V.; Mikitas, Olga V.; Egger, Denise; Lukyanov, Konstantin A.; Bienz, Kurt; Agol, Vadim I.

2004-01-01

Poliovirus and some other picornaviruses trigger relocation of certain nuclear proteins into the cytoplasm. Here, by using a protein changing its fluorescence color with time and containing a nuclear localization signal (NLS), we demonstrate that the poliovirus-triggered relocation is largely due to the exit of presynthesized nuclear protein into the cytoplasm. The leakiness of the nuclear envelope was also documented by the inability of nuclei from digitonin-permeabilized, virus-infected (but not mock-infected) cells to retain an NLS-containing derivative of green fluorescent protein (GFP). The cytoplasm-to-nucleus traffic was also facilitated during infection, as evidenced by experiments with GAPDH (glyceraldehyde-3-phosphate dehydrogenase), cyclin B1, and an NLS-lacking derivative of GFP, which are predominantly cytoplasmic in uninfected cells. Electron microscopy demonstrated that a bar-like barrier structure in the channel of the nuclear pores, seen in uninfected cells, was missing in the infected cells, giving the impression of fully open pores. Transient expression of poliovirus 2A protease also resulted in relocation of the nuclear proteins. Lysates from poliovirus-infected or 2A-expressing cells induced efflux of 3×EGFP-NLS from the nuclei of permeabilized uninfected cells. This activity was inhibited by the elastase inhibitors elastatinal and N-(methoxysuccinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloromethylketone (drugs known also to be inhibitors of poliovirus protease 2A), a caspase inhibitor zVAD(OMe), fmk, and some other protease inhibitors. These data suggest that 2A elicited nuclear efflux, possibly in cooperation with a zVAD(OMe).fmk-sensitive protease. However, poliovirus infection facilitated nuclear protein efflux also in cells deficient in caspase-3 and caspase-9, suggesting that the efflux may occur without the involvement of these enzymes. The biological relevance of nucleocytoplasmic traffic alterations in infected cells is discussed

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kuan, Man I; O’Dowd, John M.; Chughtai, Kamila

2016-10-15

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, withmore » 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. -- Highlights: •The majority of p53KO cells release fewer functional virions than wt cells. •Nucleocapsids do not efficiently exit the nucleus in p53KO cells. •Infoldings of the inner nuclear membrane are not efficiently formed in p53KO cells. •Cytoplasmic capsids are not efficiently re-enveloped in p53KO cells. •Reintroduction of p53 largely ameliorates these phenotypes.« less

Localization of the putative precursor of Alzheimer's disease-specific amyloid at nuclear envelopes of adult human muscle.

PubMed Central

Zimmermann, K; Herget, T; Salbaum, J M; Schubert, W; Hilbich, C; Cramer, M; Masters, C L; Multhaup, G; Kang, J; Lemaire, H G

1988-01-01

Cloning and sequence analysis revealed the putative amyloid A4 precursor (pre-A4) of Alzheimer's disease to have characteristics of a membrane-spanning glycoprotein. In addition to brain, pre-A4 mRNA was found in adult human muscle and other tissues. We demonstrate by in situ hybridization that pre-A4 mRNA is present in adult human muscle, in cultured human myoblasts and myotubes. Immunofluorescence with antipeptide antibodies shows the putative pre-A4 protein to be expressed in adult human muscle and associated with some but not all nuclear envelopes. Despite high levels of a single 3.5-kb pre-A4 mRNA species in cultured myoblasts and myotubes, the presence of putative pre-A4 protein could not be detected by immunofluorescence. This suggests that putative pre-A4 protein is stabilized and therefore functioning in the innervated muscle tissue but not in developing, i.e. non-innervated cultured muscle cells. The selective localization of the protein on distinct nuclear envelopes could reflect an interaction with motor endplates. Images PMID:2896589

Kid-mediated chromosome compaction ensures proper nuclear envelope formation.

PubMed

Ohsugi, Miho; Adachi, Kenjiro; Horai, Reiko; Kakuta, Shigeru; Sudo, Katsuko; Kotaki, Hayato; Tokai-Nishizumi, Noriko; Sagara, Hiroshi; Iwakura, Yoichiro; Yamamoto, Tadashi

2008-03-07

Toward the end of mitosis, neighboring chromosomes gather closely to form a compact cluster. This is important for reassembling the nuclear envelope around the entire chromosome mass but not individual chromosomes. By analyzing mice and cultured cells lacking the expression of chromokinesin Kid/kinesin-10, we show that Kid localizes to the boundaries of anaphase and telophase chromosomes and contributes to the shortening of the anaphase chromosome mass along the spindle axis. Loss of Kid-mediated anaphase chromosome compaction often causes the formation of multinucleated cells, specifically at oocyte meiosis II and the first couple of mitoses leading to embryonic death. In contrast, neither male meiosis nor somatic mitosis after the morula-stage is affected by Kid deficiency. These data suggest that Kid-mediated anaphase/telophase chromosome compaction prevents formation of multinucleated cells. This protection is especially important during the very early stages of development, when the embryonic cells are rich in ooplasm.

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.

Permeabilization of the nuclear envelope following nanosecond pulsed electric field exposure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thompson, Gary L., E-mail: gary.l.thompson.3@gmail.com; Roth, Caleb C.; Kuipers, Marjorie A.

2016-01-29

Permeabilization of cell membranes occurs upon exposure to a threshold absorbed dose (AD) of nanosecond pulsed electric fields (nsPEF). The ultimate, physiological bioeffect of this exposure depends on the type of cultured cell and environment, indicating that cell-specific pathways and structures are stimulated. Here we investigate 10 and 600 ns duration PEF effects on Chinese hamster ovary (CHO) cell nuclei, where our hypothesis is that pulse disruption of the nuclear envelope membrane leads to observed cell death and decreased viability 24 h post-exposure. To observe short-term responses to nsPEF exposure, CHO cells have been stably transfected with two fluorescently-labeled proteins known tomore » be sequestered for cellular chromosomal function within the nucleus – histone-2b (H2B) and proliferating cell nuclear antigen (PCNA). H2B remains associated with chromatin after nsPEF exposure, whereas PCNA leaks out of nuclei permeabilized by a threshold AD of 10 and 600 ns PEF. A downturn in 24 h viability, measured by MTT assay, is observed at the number of pulses required to induce permeabilization of the nucleus. - Highlights: • The ability of nsPEF to damage nuclear structures within cells is investigated. • Leakage of proliferating nuclear antigen from nuclei is induced by nsPEF. • High doses of nsPEF disrupt cortical lamin and cause chromatin decompaction. • Histone H2B remains attached to chromatin following nsPEF exposure. • DNA does not leak out of nsPEF-permeabilized nuclei.« less

Permeabilization of the nuclear envelope following nanosecond pulsed electric field exposure.

PubMed

Thompson, Gary L; Roth, Caleb C; Kuipers, Marjorie A; Tolstykh, Gleb P; Beier, Hope T; Ibey, Bennett L

2016-01-29

Permeabilization of cell membranes occurs upon exposure to a threshold absorbed dose (AD) of nanosecond pulsed electric fields (nsPEF). The ultimate, physiological bioeffect of this exposure depends on the type of cultured cell and environment, indicating that cell-specific pathways and structures are stimulated. Here we investigate 10 and 600 ns duration PEF effects on Chinese hamster ovary (CHO) cell nuclei, where our hypothesis is that pulse disruption of the nuclear envelope membrane leads to observed cell death and decreased viability 24 h post-exposure. To observe short-term responses to nsPEF exposure, CHO cells have been stably transfected with two fluorescently-labeled proteins known to be sequestered for cellular chromosomal function within the nucleus - histone-2b (H2B) and proliferating cell nuclear antigen (PCNA). H2B remains associated with chromatin after nsPEF exposure, whereas PCNA leaks out of nuclei permeabilized by a threshold AD of 10 and 600 ns PEF. A downturn in 24 h viability, measured by MTT assay, is observed at the number of pulses required to induce permeabilization of the nucleus. Copyright © 2015 Elsevier Inc. All rights reserved.

Nuclear envelope breakdown and mitosis in sand dollar embryos is inhibited by microinjection of calcium buffers in a calcium-reversible fashion, and by antagonists of intracellular Ca2+ channels.

PubMed

Silver, R B

1989-01-01

Transient elevations in intracellular free Ca2+ are believed to signal the initiation of mitosis. This model predicts that mitosis might be arrested prior to nuclear envelope breakdown (NEB) or anaphase onset if intracellular Ca2+ concentration is buffered or dampened. Microinjection of a discrete dose of Ca2+ into the cell might then release the cell to resume mitotic cycling. Experimentally, one blastomere of two cell sand dollar (Echinaracnius parma) embryos was microinjected with Ca2+ buffers, Ca2+ solutions, or Ca2+ channel antagonists; the uninjected blastomere was the control. Cells were loaded with 10 pl doses of the Ca2+ buffer antipyrylazo III (ApIII) at specific times in the cell cycle to attempt a competitive inhibition of Ca2+-dependent steps in NEB and initiation of mitosis. Injection of 50 microM ApIII 6 min prior to NEB blocked NEB and further cell cycling. Injections of solutions between 0 and 30 microM ApIII were without observable effect. Control injections had no observable effect on the injected cell. Cells injected with 50 microM ApIII 2 min prior to the onset of anaphase in control cells were blocked in metaphase. Cells were sensitive to Ca2+ buffer injections 6 min prior to NEB (with a 40- to 45-sec duration), and 2 min prior to anaphase onset (with a 10- to 20-sec duration). Vital staining of these cells with H33342 demonstrated that they contained only one nucleus that had the same fluorescence intensity as seen prior to microinjection, and thus did not undergo DNA synthesis following the imposition of the Ca2+ buffer block to mitosis. Cells arrested in this fashion did not spontaneously resume mitotic cycling. This Ca2+ buffer-induced mitotic arrest was, however, experimentally reversible. Cells arrested with 50 microM ApIII 6 min prior to NEB could be returned to mitotic activity by injecting 300 microM CaCl2 5 min after the ApIII injection. The double injected cells resumed cycling, NEB, and mitosis after a delay of one cell cycle

Enhancing Electrotransfection Efficiency through Improvement in Nuclear Entry of Plasmid DNA.

PubMed

Cervia, Lisa D; Chang, Chun-Chi; Wang, Liangli; Mao, Mao; Yuan, Fan

2018-06-01

The nuclear envelope is a physiological barrier to electrogene transfer. To understand different mechanisms of the nuclear entry for electrotransfected plasmid DNA (pDNA), the current study investigated how manipulation of the mechanisms could affect electrotransfection efficiency (eTE), transgene expression level (EL), and cell viability. In the investigation, cells were first synchronized at G2-M phase prior to electrotransfection so that the nuclear envelope breakdown (NEBD) occurred before pDNA entered the cells. The NEBD significantly increased the eTE and the EL while the cell viability was not compromised. In the second experiment, the cells were treated with a nuclear pore dilating agent (i.e., trans-1,2-cyclohexanediol). The treatment could increase the EL, but had only minor effects on eTE. Furthermore, the treatment was more cytotoxic, compared with the cell synchronization. In the third experiment, a nuclear targeting sequence (i.e., SV40) was incorporated into the pDNA prior to electrotransfection. The incorporation was more effective than the cell synchronization for enhancing the EL, but not the eTE, and the effectiveness was cell type dependent. Taken together, the data described above suggested that synchronization of the NEBD could be a practical approach to improving electrogene transfer in all dividing cells. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Electrostatic association of glutathione transferase to the nuclear membrane. Evidence of an enzyme defense barrier at the nuclear envelope.

PubMed

Stella, Lorenzo; Pallottini, Valentina; Moreno, Sandra; Leoni, Silvia; De Maria, Francesca; Turella, Paola; Federici, Giorgio; Fabrini, Raffaele; Dawood, Kutayba F; Bello, Mario Lo; Pedersen, Jens Z; Ricci, Giorgio

2007-03-02

The possible nuclear compartmentalization of glutathione S-transferase (GST) isoenzymes has been the subject of contradictory reports. The discovery that the dinitrosyl-diglutathionyl-iron complex binds tightly to Alpha class GSTs in rat hepatocytes and that a significant part of the bound complex is also associated with the nuclear fraction (Pedersen, J. Z., De Maria, F., Turella, P., Federici, G., Mattei, M., Fabrini, R., Dawood, K. F., Massimi, M., Caccuri, A. M., and Ricci, G. (2007) J. Biol. Chem. 282, 6364-6371) prompted us to reconsider the nuclear localization of GSTs in these cells. Surprisingly, we found that a considerable amount of GSTs corresponding to 10% of the cytosolic pool is electrostatically associated with the outer nuclear membrane, and a similar quantity is compartmentalized inside the nucleus. Mainly Alpha class GSTs, in particular GSTA1-1, GSTA2-2, and GSTA3-3, are involved in this double modality of interaction. Confocal microscopy, immunofluorescence experiments, and molecular modeling have been used to detail the electrostatic association in hepatocytes and liposomes. A quantitative analysis of the membrane-bound Alpha GSTs suggests the existence of a multilayer assembly of these enzymes at the outer nuclear envelope that could represent an amazing novelty in cell physiology. The interception of potentially noxious compounds to prevent DNA damage could be the possible physiological role of the perinuclear and intranuclear localization of Alpha GSTs.

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)

Dual roles of TRF1 in tethering telomeres to the nuclear envelope and protecting them from fusion during meiosis.

PubMed

Wang, Lina; Tu, Zhaowei; Liu, Chao; Liu, Hongbin; Kaldis, Philipp; Chen, Zijiang; Li, Wei

2018-06-01

Telomeres integrity is indispensable for chromosomal stability by preventing chromosome erosion and end-to-end fusions. During meiosis, telomeres attach to the inner nuclear envelope and cluster into a highly crowded microenvironment at the bouquet stage, which requires specific mechanisms to protect the telomeres from fusion. Here, we demonstrate that germ cell-specific knockout of a shelterin complex subunit, Trf1, results in arrest of spermatocytes at two different stages. The obliterated telomere-nuclear envelope attachment in Trf1-deficient spermatocytes impairs homologue synapsis and recombination, resulting in a pachytene-like arrest, while the meiotic division arrest might stem from chromosome end-to-end fusion due to the failure of recruiting meiosis specific telomere associated proteins. Further investigations uncovered that TRF1 could directly interact with Speedy A, and Speedy A might work as a scaffold protein to further recruit Cdk2, thus protecting telomeres from fusion at this stage. Together, our results reveal a novel mechanism of TRF1, Speedy A, and Cdk2 in protecting telomere from fusion in a highly crowded microenvironment during meiosis.

Full-Waveform Envelope Templates for Low Magnitude Discrimination and Yield Estimation at Local and Regional Distances with Application to the North Korean Nuclear Tests

NASA Astrophysics Data System (ADS)

Yoo, S. H.

2017-12-01

Monitoring seismologists have successfully used seismic coda for event discrimination and yield estimation for over a decade. In practice seismologists typically analyze long-duration, S-coda signals with high signal-to-noise ratios (SNR) at regional and teleseismic distances, since the single back-scattering model reasonably predicts decay of the late coda. However, seismic monitoring requirements are shifting towards smaller, locally recorded events that exhibit low SNR and short signal lengths. To be successful at characterizing events recorded at local distances, we must utilize the direct-phase arrivals, as well as the earlier part of the coda, which is dominated by multiple forward scattering. To remedy this problem, we have developed a new hybrid method known as full-waveform envelope template matching to improve predicted envelope fits over the entire waveform and account for direct-wave and early coda complexity. We accomplish this by including a multiple forward-scattering approximation in the envelope modeling of the early coda. The new hybrid envelope templates are designed to fit local and regional full waveforms and produce low-variance amplitude estimates, which will improve yield estimation and discrimination between earthquakes and explosions. To demonstrate the new technique, we applied our full-waveform envelope template-matching method to the six known North Korean (DPRK) underground nuclear tests and four aftershock events following the September 2017 test. We successfully discriminated the event types and estimated the yield for all six nuclear tests. We also applied the same technique to the 2015 Tianjin explosions in China, and another suspected low-yield explosion at the DPRK test site on May 12, 2010. Our results show that the new full-waveform envelope template-matching method significantly improves upon longstanding single-scattering coda prediction techniques. More importantly, the new method allows monitoring seismologists to extend

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.

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.

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

UL31 and UL34 Proteins of Herpes Simplex Virus Type 1 Form a Complex That Accumulates at the Nuclear Rim and Is Required for Envelopment of Nucleocapsids

PubMed Central

Reynolds, Ashley E.; Ryckman, Brent J.; Baines, Joel D.; Zhou, Yuping; Liang, Li; Roller, Richard J.

2001-01-01

The herpes simplex virus type 1 (HSV-1) UL34 protein is likely a type II membrane protein that localizes within the nuclear membrane and is required for efficient envelopment of progeny virions at the nuclear envelope, whereas the UL31 gene product of HSV-1 is a nuclear matrix-associated phosphoprotein previously shown to interact with UL34 protein in HSV-1-infected cell lysates. For these studies, polyclonal antisera directed against purified fusion proteins containing UL31 protein fused to glutathione-S-transferase (UL31-GST) and UL34 protein fused to GST (UL34-GST) were demonstrated to specifically recognize the UL31 and UL34 proteins of approximately 34,000 and 30,000 Da, respectively. The UL31 and UL34 gene products colocalized in a smooth pattern throughout the nuclear rim of infected cells by 10 h postinfection. UL34 protein also accumulated in pleiomorphic cytoplasmic structures at early times and associated with an altered nuclear envelope late in infection. Localization of UL31 protein at the nuclear rim required the presence of UL34 protein, inasmuch as cells infected with a UL34 null mutant virus contained UL31 protein primarily in central intranuclear domains separate from the nuclear rim, and to a lesser extent in the cytoplasm. Conversely, localization of UL34 protein exclusively at the nuclear rim required the presence of the UL31 gene product, inasmuch as UL34 protein was detectable at the nuclear rim, in replication compartments, and in the cytoplasm of cells infected with a UL31 null virus. When transiently expressed in the absence of other viral factors, UL31 protein localized diffusely in the nucleoplasm, whereas UL34 protein localized primarily in the cytoplasm and at the nuclear rim. In contrast, coexpression of the UL31 and UL34 proteins was sufficient to target both proteins exclusively to the nuclear rim. The proteins were also shown to directly interact in vitro in the absence of other viral proteins. In cells infected with a virus

Analysis of simulated high burnup nuclear fuel by laser induced breakdown spectroscopy

NASA Astrophysics Data System (ADS)

Singh, Manjeet; Sarkar, Arnab; Banerjee, Joydipta; Bhagat, R. K.

2017-06-01

Advanced Heavy Water Reactor (AHWR) grade (Th-U)O2 fuel sample and Simulated High Burn-Up Nuclear Fuels (SIMFUEL) samples mimicking the 28 and 43 GWd/Te irradiated burn-up fuel were studied using laser-induced breakdown spectroscopy (LIBS) setup in a simulated hot-cell environment from a distance of > 1.5 m. Resolution of < 38 pm has been used to record the complex spectra of the SIMFUEL samples. By using spectrum comparison and database matching > 60 emission lines of fission products was identified. Among them only a few emission lines were found to generate calibration curves. The study demonstrates the possibility to investigate impurities at concentrations around hundreds of ppm, rapidly at atmospheric pressure without any sample preparation. The results of Ba and Mo showed the advantage of LIBS analysis over traditional methods involving sample dissolution, which introduces possible elemental loss. Limits of detections (LOD) under Ar atmosphere shows significant improvement, which is shown to be due to the formation of stable plasma.

Transcriptional repression mediated by repositioning of genes to the nuclear lamina.

PubMed

Reddy, K L; Zullo, J M; Bertolino, E; Singh, H

2008-03-13

Nuclear compartmentalization seems to have an important role in regulating metazoan genes. Although studies on immunoglobulin and other loci have shown a correlation between positioning at the nuclear lamina and gene repression, the functional consequences of this compartmentalization remain untested. We devised an approach for inducible tethering of genes to the inner nuclear membrane (INM), and tested the consequences of such repositioning on gene activity in mouse fibroblasts. Here, using three-dimensional DNA-immunoFISH, we demonstrate repositioning of chromosomal regions to the nuclear lamina that is dependent on breakdown and reformation of the nuclear envelope during mitosis. Moreover, tethering leads to the accumulation of lamin and INM proteins, but not to association with pericentromeric heterochromatin or nuclear pore complexes. Recruitment of genes to the INM can result in their transcriptional repression. Finally, we use targeted adenine methylation (DamID) to show that, as is the case for our model system, inactive immunoglobulin loci at the nuclear periphery are contacted by INM and lamina proteins. We propose that these molecular interactions may be used to compartmentalize and to limit the accessibility of immunoglobulin loci to transcription and recombination factors.

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. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

Analysis of codon usage bias of envelope glycoprotein genes in nuclear polyhedrosis virus (NPV) and its relation to evolution.

PubMed

Zhao, Yongchao; Zheng, Hao; Xu, Anying; Yan, Donghua; Jiang, Zijian; Qi, Qi; Sun, Jingchen

2016-08-24

Analysis of codon usage bias is an extremely versatile method using in furthering understanding of the genetic and evolutionary paths of species. Codon usage bias of envelope glycoprotein genes in nuclear polyhedrosis virus (NPV) has remained largely unexplored at present. Hence, the codon usage bias of NPV envelope glycoprotein was analyzed here to reveal the genetic and evolutionary relationships between different viral species in baculovirus genus. A total of 9236 codons from 18 different species of NPV of the baculovirus genera were used to perform this analysis. Glycoprotein of NPV exhibits weaker codon usage bias. Neutrality plot analysis and correlation analysis of effective number of codons (ENC) values indicate that natural selection is the main factor influencing codon usage bias, and that the impact of mutation pressure is relatively smaller. Another cluster analysis shows that the kinship or evolutionary relationships of these viral species can be divided into two broad categories despite all of these 18 species are from the same baculovirus genus. There are many elements that can affect codon bias, such as the composition of amino acids, mutation pressure, natural selection, gene expression level, and etc. In the meantime, cluster analysis also illustrates that codon usage bias of virus envelope glycoprotein can serve as an effective means of evolutionary classification in baculovirus genus.

Three-dimensional visualization of the Autographa californica multiple nucleopolyhedrovirus occlusion-derived virion envelopment process gives new clues as to its mechanism

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shi, Yang; Li, Kunpeng; Tang, Peiping

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 inmore » 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.« less

Identification of a major polypeptide of the nuclear pore complex

PubMed Central

1982-01-01

The nuclear pore complex is a prominent structural component of the nuclear envelope that appears to regulate nucleoplasmic molecular movement. Up to now, none of its polypeptides have been defined. To identify possible pore complex proteins, we fractionated rat liver nuclear envelopes and microsomal membranes with strong protein perturbants into peripheral and intrinsic membrane proteins, and compared these fractions on SDS gels. From this analysis, we identified a prominent 190-kilodalton intrinsic membrane polypeptide that occurs specifically in nuclear envelopes. Lectin binding studies indicate that this polypeptide (gp 190) is the major nuclear envelope glycoprotein. Upon treatment of nuclear envelopes with Triton X-100, gp 190 remains associated with a protein substructure of the nuclear envelope consisting of pore complexes and nuclear lamina. We prepared monospecific antibodies to gp 190 for immunocytochemical localization. Immunofluorescence staining of tissue culture cells suggests that gp 190 occurs exclusively in the nucleus during interphase. This polypeptide becomes dispersed throughout the cell in mitotic prophase when the nuclear envelope is disassembled, and subsequently returns to the nuclear surfaces during telophase when the nuclear envelope is reconstructed. Immunoferritin labeling of Triton-treated rat liver nuclei demonstrates that gp 190 occurs exclusively in the nuclear pore complex, in the regions of the cytoplasmic (and possibly nucleoplasmic) pore complex annuli. A polypeptide that cross-reacts with gp 190 is present in diverse vertebrate species, as shown by antibody labeling of nitrocellulose SDS gel transfers. On the basis of its biochemical characteristics, we suggest that gp 190 may be involved in anchoring the pore complex to nuclear envelope membranes. PMID:7153248

Mitotic phosphorylation of SUN1 loosens its connection with the nuclear lamina while the LINC complex remains intact.

PubMed

Patel, Jennifer T; Bottrill, Andrew; Prosser, Suzanna L; Jayaraman, Sangeetha; Straatman, Kees; Fry, Andrew M; Shackleton, Sue

2014-01-01

At the onset mitosis in higher eukaryotes, the nuclear envelope (NE) undergoes dramatic deconstruction to allow separation of duplicated chromosomes. Studies have shown that during this process of nuclear envelope breakdown (NEBD), the extensive protein networks of the nuclear lamina are disassembled through phosphorylation of lamins and several inner nuclear membrane (INM) proteins. The LINC complex, composed of SUN and nesprin proteins, is involved in multiple interactions at the NE and plays vital roles in nuclear and cellular mechanics by connecting the nucleus to the cytoskeleton. Here, we show that SUN1, located in the INM, undergoes mitosis-specific phosphorylation on at least 3 sites within its nucleoplasmic N-terminus. We further identify Cdk1 as the kinase responsible for serine 48 and 333 phosphorylation, while serine 138 is phosphorylated by Plk1. In mitotic cells, SUN1 loses its interaction with N-terminal domain binding partners lamin A/C, emerin, and short nesprin-2 isoforms. Furthermore, a triple phosphomimetic SUN1 mutant displays increased solubility and reduced retention at the NE. In contrast, the central LINC complex interaction between the SUN1 C-terminus and the KASH domain of nesprin-2 is maintained during mitosis. Together, these data support a model whereby mitotic phosphorylation of SUN1 disrupts interactions with nucleoplasmic binding partners, promoting disassembly of the nuclear lamina and, potentially, its chromatin interactions. At the same time, our data add to an emerging picture that the core LINC complex plays an active role in NEBD.

Luteinizing hormone-accelerated redistribution of lysosome-like organelles preceding dissolution of the nuclear envelope in rat oocytes maturing in vitro

PubMed Central

1979-01-01

Maturation of the mammalian oocyte is characterized in part by dissolution of the nuclear envelope, or germinal vesicle breakdown (GVB). By fluorescence microscopy after vital uptake of acridine orange (AO), redistribution and perinuclear accumulation of organelles corresponding to lysosomes occur before GVB in rat oocytes undergoing meiotic maturation in vitro. In follicle-enclosed oocytes explanted during the preovulatory gonadotropin surge (GS) and individually cultured as such in chemically defined medium at approximately 22 degrees C, lysosomes aggregated into disperse clusters after 30 min; by 60 min, perinuclear concentration of lysosomes and their essential disappearance from the cortical ooplasm were observed. GVB occurred within 120 min. In contrast, follicle-enclosed oocytes explanted before the GS displayed a generally homogeneous distribution of lysosomes and intact GV for up to 5 h in culture. In oocytes aspirated from follicles before the GS, partially denuded of granulosa cells, and cultivated without added hormone, most lysosomes concentrated around the GV within 60 min, with GVB occurring generally by 120 min. Luteinizing hormone (LH) added in vitro to the isolated preparation at 3 or 30 x 10(-8) M sharply accelerated these events. The effects of LH, not seen with 1.5 x 10(-8) M hormone, were blocked by anti-LH IgG. Up to 60 x 10(-8) M follicle-stimulating hormone or 80 x 10(-8) M prolactin were ineffective in accelerating lysosome redistribution or GVB. After GVB, lysosomes became once again uniformly dispersed and unresponsive, even to 60 x 10(-8) M added LH, a finding consistent with tachyphylaxis of target cells by independent criteria. The present data, all statistically significant at P less than 0.05, demonstrate that mobilization of lysosomes before GVB is a specific response to factors that promote resumption of meiotic maturation of rat oocytes. PMID:573271

Nesprin-1α-Dependent Microtubule Nucleation from the Nuclear Envelope via Akap450 Is Necessary for Nuclear Positioning in Muscle Cells.

PubMed

Gimpel, Petra; Lee, Yin Loon; Sobota, Radoslaw M; Calvi, Alessandra; Koullourou, Victoria; Patel, Rutti; Mamchaoui, Kamel; Nédélec, François; Shackleton, Sue; Schmoranzer, Jan; Burke, Brian; Cadot, Bruno; Gomes, Edgar R

2017-10-09

The nucleus is the main microtubule-organizing center (MTOC) in muscle cells due to the accumulation of centrosomal proteins and microtubule (MT) nucleation activity at the nuclear envelope (NE) [1-4]. The relocalization of centrosomal proteins, including Pericentrin, Pcm1, and γ-tubulin, depends on Nesprin-1, an outer nuclear membrane (ONM) protein that connects the nucleus to the cytoskeleton via its N-terminal region [5-7]. Nesprins are also involved in the recruitment of kinesin to the NE and play a role in nuclear positioning in skeletal muscle cells [8-12]. However, a function for MT nucleation from the NE in nuclear positioning has not been established. Using the proximity-dependent biotin identification (BioID) method [13, 14], we found several centrosomal proteins, including Akap450, Pcm1, and Pericentrin, whose association with Nesprin-1α is increased in differentiated myotubes. We show that Nesprin-1α recruits Akap450 to the NE independently of kinesin and that Akap450, but not other centrosomal proteins, is required for MT nucleation from the NE. Furthermore, we demonstrate that this mechanism is disrupted in congenital muscular dystrophy patient myotubes carrying a nonsense mutation within the SYNE1 gene (23560 G>T) encoding Nesprin-1 [15, 16]. Finally, using computer simulation and cell culture systems, we provide evidence for a role of MT nucleation from the NE on nuclear spreading in myotubes. Our data thus reveal a novel function for Nesprin-1α/Nesprin-1 in nuclear positioning through recruitment of Akap450-mediated MT nucleation activity to the NE. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

SEPT12/SPAG4/LAMINB1 complexes are required for maintaining the integrity of the nuclear envelope in postmeiotic male germ cells.

PubMed

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

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.

Influence of acute promyelocytic leukemia therapeutic drugs on nuclear pore complex density and integrity.

PubMed

Lång, Anna; Øye, Alexander; Eriksson, Jens; Rowe, Alexander D; Lång, Emma; Bøe, Stig Ove

2018-05-15

During cell division, a large number of nuclear proteins are released into the cytoplasm due to nuclear envelope breakdown. Timely nuclear import of these proteins following exit from mitosis is critical for establishment of the G1 nuclear environment. Dysregulation of post-mitotic nuclear import may affect the fate of newly divided stem or progenitor cells and may lead to cancer. Acute promyelocytic leukemia (APL) is a malignant disorder that involves a defect in blood cell differentiation at the promyelocytic stage. Recent studies suggest that pharmacological concentrations of the APL therapeutic drugs, all-trans retinoic acid (ATRA) and arsenic trioxide (ATO), affect post-mitotic nuclear import of the APL-associated oncoprotein PML/RARA. In the present study, we have investigated the possibility that ATRA and ATO affect post-mitotic nuclear import through interference with components of the nuclear import machinery. We observe reduced density and impaired integrity of nuclear pore complexes after ATRA and/or ATO exposure. Using a post-mitotic nuclear import assay, we demonstrate distinct import kinetics among different nuclear import pathways while nuclear import rates were similar in the presence or absence of APL therapeutic drugs. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

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

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.

Enveloping Aerodynamic Decelerator

NASA Technical Reports Server (NTRS)

Nock, Kerry T. (Inventor); Aaron, Kim M. (Inventor); McRonald, Angus D. (Inventor); Gates, Kristin L. (Inventor)

2018-01-01

An inflatable aerodynamic deceleration method and system is provided for use with an atmospheric entry payload. The inflatable aerodynamic decelerator includes an inflatable envelope and an inflatant, wherein the inflatant is configured to fill the inflatable envelope to an inflated state such that the inflatable envelope surrounds the atmospheric entry payload, causing aerodynamic forces to decelerate the atmospheric entry payload.

A baculovirus polyhedron envelope protein: immunogold localization in infected cells and mature polyhedra.

PubMed

Russell, R L; Rohrmann, G F

1990-01-01

A polyclonal antiserum against a trpE fusion protein containing the complete open reading frame of the polyhedron envelope (PE) protein from the nuclear polyhedrosis virus of Orgyia pseudotsugata (OpMNPV) was used for immunogold staining and electron microscopic examination of polyhedra, isolated polyhedron envelopes, and infected insect cells at selected times postinfection. The antiserum specifically stained the peripheral envelope of mature polyhedra and also stained the envelope structure which remained after polyhedra were dissolved in dilute alkaline solutions. In OpMNPV-infected Lymantria dispar cells, the PE protein was detected by 48 hr postinfection (hr p.i.) but specific localization and staining of developing polyhedra were not evident. However, by 72 hr p.i. substantial and preferential staining of the periphery of developing polyhedra was evident even though a distinct polyhedron envelope was not yet observed. In addition, the periphery of fibrillar structures was stained by the PE antiserum. By 96 hr p.i., mature envelopes surrounded polyhedra and these polyhedron envelopes were stained with the PE antibody. The progression of PE protein staining during polyhedron morphogenesis indicates that the PE protein accumulates and becomes associated with developing polyhedra in the nucleus between 48 and 72 hr p.i. Very late in infection the mature polyhedron envelope forms on the polyhedron surface. The apparent affinity of the PE protein for the surface of maturing polyhedra suggests that it may be a major component of the polyhedron envelope or may form the matrix for the deposition of other components which contribute to the mature envelope. Immunogold staining and protease digestion experiments indicate that protein is an essential component of the polyhedron envelope.

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

Early morphological nuclear events and developmental capacity of embryos reconstructed with fetal fibroblasts at the M or G1 phase after intracytoplasmic nuclear injection in cattle.

PubMed

Ideta, Atsushi; Urakawa, Manami; Aoyagi, Yoshito; Saeki, Kazuhiro

2005-04-01

We examined morphological nuclear events during the first cell cycle of bovine embryos reconstructed with somatic cells at the M and G1 phases (M-embryos and G1-embryos, respectively) by intracytoplasmic nuclear injection, and the subsequent development of these embryos in vitro and in vivo. Bovine fetal fibroblasts (BFFs) at the M or G1 phase were directly injected into enucleated oocytes, and activated immediately. Only half (48%) of the M-embryos extruded polar body-like cells (PBCs) at 6 h post injection (hpi). At 15 to 19 hpi, 54% of the M-embryos formed a single pronucleus-like nucleus. Nuclear envelope-breakdown, premature chromosome condensation and single nuclear clusters were observed in most of the G1-embryos (88%) within 30 min following the nuclear injection. At 15 to 19 hpi, single pronucleus-like nuclei were formed in most G1-embryos (83%). The potential of G1-embryos to develop to blastocysts was significantly higher than that of M-embryos (31% vs 16%). Three of five recipients following transfer of blastocysts derived from the G1-embryos became pregnant on Day 30, and one recipient delivered a calf. Our results indicate that almost a half of the M-embryos failed to extrude PBCs and that the G1-embryos developed to blastocysts at a higher rate than the M-embryos.

Membrane and Nuclear Permeabilization by Polymeric pDNA Vehicles: Efficient Method for Gene Delivery or Mechanism of Cytotoxicity?

PubMed Central

Grandinetti, Giovanna; Smith, Adam E.; Reineke, Theresa M.

2012-01-01

The aim of this study is to compare the cytotoxicity mechanisms of linear PEI to two analogous polymers synthesized by our group: a hydroxyl-containing poly(L-tartaramidoamine) (T4) and a version containing an alkyl chain spacer poly(adipamidopentaethylenetetramine) (A4) by studying the cellular responses to polymer transfection. We have also synthesized analogues of T4 with different molecular weights (degrees of polymerization of 6, 12, and 43) to examine the role of molecular weight on the cytotoxicity mechanisms. Several mechanisms of polymer-induced cytotoxicity are investigated, including plasma membrane permeabilization, the formation of potentially harmful polymer degradation products during transfection including reactive oxygen species, and nuclear membrane permeabilization. We hypothesized that since cationic polymers are capable of disrupting the plasma membrane, they may also be capable of disrupting the nuclear envelope, which could be a potential mechanism of how the pDNA is delivered into the nucleus (other than nuclear envelope breakdown during mitosis). Using flow cytometry and confocal microscopy, we show that the polycations with the highest amount of protein expression and toxicity, PEI and T443, are capable of inducing nuclear membrane permeability. This finding is important for the field of nucleic acid delivery in that not only could direct nucleus permeabilization be a mechanism for pDNA nuclear import but also a potential mechanism of cytotoxicity and cell death. We also show that the production of reactive oxygen species is not a main mechanism of cytotoxicity, and that the presence or absence of hydroxyl groups as well as polymer length plays a role in polyplex size and charge in addition to protein expression efficiency and toxicity. PMID:22175236

Multi-scale signed envelope inversion

NASA Astrophysics Data System (ADS)

Chen, Guo-Xin; Wu, Ru-Shan; Wang, Yu-Qing; Chen, Sheng-Chang

2018-06-01

Envelope inversion based on modulation signal mode was proposed to reconstruct large-scale structures of underground media. In order to solve the shortcomings of conventional envelope inversion, multi-scale envelope inversion was proposed using new envelope Fréchet derivative and multi-scale inversion strategy to invert strong contrast models. In multi-scale envelope inversion, amplitude demodulation was used to extract the low frequency information from envelope data. However, only to use amplitude demodulation method will cause the loss of wavefield polarity information, thus increasing the possibility of inversion to obtain multiple solutions. In this paper we proposed a new demodulation method which can contain both the amplitude and polarity information of the envelope data. Then we introduced this demodulation method into multi-scale envelope inversion, and proposed a new misfit functional: multi-scale signed envelope inversion. In the numerical tests, we applied the new inversion method to the salt layer model and SEG/EAGE 2-D Salt model using low-cut source (frequency components below 4 Hz were truncated). The results of numerical test demonstrated the effectiveness of this method.

Arabidopsis WPP-Domain Proteins Are Developmentally Associated with the Nuclear Envelope and Promote Cell DivisionW⃞

PubMed Central

Patel, Shalaka; Rose, Annkatrin; Meulia, Tea; Dixit, Ram; Cyr, Richard J.; Meier, Iris

2004-01-01

The nuclear envelope (NE) acts as a selective barrier to macromolecule trafficking between the nucleus and the cytoplasm and undergoes a complex reorganization during mitosis. Different eukaryotic kingdoms show specializations in NE function and composition. In contrast with vertebrates, the protein composition of the NE and the function of NE proteins are barely understood in plants. MFP1 attachment factor 1 (MAF1) is a plant-specific NE-associated protein first identified in tomato (Lycopersicon esculentum). Here, we demonstrate that two Arabidopsis thaliana MAF1 homologs, WPP1 and WPP2, are associated with the NE specifically in undifferentiated cells of the root tip. Reentry into cell cycle after callus induction from differentiated root segments reprograms their NE association. Based on green fluorescent protein fusions and immunogold labeling data, the proteins are associated with the outer NE and the nuclear pores in interphase cells and with the immature cell plate during cytokinesis. RNA interference–based suppression of the Arabidopsis WPP family causes shorter primary roots, a reduced number of lateral roots, and reduced mitotic activity of the root meristem. Together, these data demonstrate the existence of regulated NE targeting in plants and identify a class of plant-specific NE proteins involved in mitotic activity. PMID:15548735

Histone-poly(A) hybrid molecules as tools to block nuclear pores.

PubMed

Cremer, G; Wojtech, E; Kalbas, M; Agutter, P S; Prochnow, D

1995-04-01

Histone-poly(A) hybrid molecules were used for transport experiments with resealed nuclear envelopes and after attachment of a cleavable cross-linker (SASD) to identify nuclear proteins. In contrast to histones, the hybrid molecules cannot be accumulated in resealed nuclear envelopes, and in contrast to poly(A), the export of hybrids from preloaded nuclear envelopes is completely impaired. The experiments strongly confirm the existence of poly(A) as an export signal in mRNA which counteracts the nuclear location signals (NLS) in histones. The contradicting transport signals in the hybrid molecules impair translocation through the nuclear pore complex. The failure to accumulate hybrid molecules into resealed nuclear envelopes results from the covalent attachment of polyadenylic acid to histones in a strict 1:1 molar ratio. This was demonstrated in control transport experiments where radiolabeled histones were simply mixed with nonlabeled poly(A) or radiolabeled poly(A) mixed with nonlabeled histones. In comparison, control uptake experiments with histones covalently linked to a single UMP-mononucleotide are strongly enhanced. Such controls exclude the conceivable possibility of a simple masking of the nuclear location signal in the histones by the covalent attached poly(A) moiety. Photoreactive histone-poly(A) hybrid analogs serve to identify nuclear envelope proteins--presumably in the nuclear pore--with molecular weights of 110, 80, and 71.4 kDa.

AC BREAKDOWN IN GASES

DTIC Science & Technology

electron- emission (multipactor) region, and (3) the low-frequency region. The breakdown mechanism in each of these regions is explained. An extensive bibliography on AC breakdown in gases is included.

Electrophoretic characterization of the Mammalian nuclear matrix proteome, nuclear envelope, nucleoli and covalently bound ADP-ribose polymers: potential applications to cancer.

PubMed

Aranda, Xavier G; Racho, Ronald G; Pacheco-Rodríguez, Gustavo; Alvarez-González, Rafael

2014-01-01

Nucleic acid metabolism is biochemically compartmentalized to the nucleus. Thus, it is necessary to define the proteome of the various macromolecular structures within this organelle. We isolated the nuclear matrix (NM) fraction from rat liver by sequential centrifugation steps at 13,000 rpm, staggered between endogenous nuclease treatment for 2 h at 37°C, followed by high-salt (H.S.; 2.0 M NaCl) and non-ionic detergent extractions (0.1%- or 1.0% Triton X-100) to eliminate the bulk of chromosomal DNA/RNA, histone proteins and the nuclear envelope (NE). Integrity of the NM and NE structures was confirmed by electron microscopy. Next, we analyzed the NM proteome on a 20% polyacrylamide gel using the PhastSystem. We observed the absence of histone proteins and the characteristic presence of the lamins by Coomassie blue staining. By contrast, upon silver staining, following electrophoretic separation with a Tris-Borate-EDTA buffer, we observed the NM-associated nucleic RNA and protein-free ADP-ribose polymers. While polymers are found in much lower concentration than RNA in NM, they were purified by affinity chromatography on boronate resin prior to electrophoresis. We observed the electrophoretic resolution of free ADP-ribose chains (5-25 units) by silver staining. The significance of our observations to cancer studies and carcinogenesis is discussed. Copyright© 2014, International Institute of Anticancer Research (Dr. John G. Delinasios), All rights reserved.

["Nervous breakdown": a diagnostic characterization study].

PubMed

Salmán, E; Carrasco, J L; Liebowitz, M; Díaz Marsá, M; Prieto, R; Jusino, C; Cárdenas, D; Klein, D

1997-01-01

An evaluation was made of the influence of different psychiatric co-morbidities on the symptoms of the disorder popularly known as "ataque de nervios" (nervous breakdown) among the US Hispanic population. Using a self-completed instrument designed specially for both traditional nervous breakdown and for panic symptoms, and structured or semi-structured psychiatric interviews for Axis I disorders, and evaluation was made of Hispanic subjects who sought treatment for anxiety in a clinic (n = 156). This study centered on 102 subjects who presented symptoms of "nervous breakdown" and comorbidity with panic disorder, other anxiety disorders, or affective disorder. Variations in co-morbidity with "nervous breakdown" enabled the identification of different patterns of "nervous breakdown" presenting symptoms. Individuals with "nervous breakdown" and panic disorder characteristically expressed a greater sense of asphyxiation, fear of dying, and growing fear (panic-like) during their breakdowns. Subjects with "nervous breakdown" and affective disorder had a greater sensation of anger and more tendency toward screaming and aggressive behavior such as breaking things during the breakdown (emotional anger). Finally, subjects with "nervous breakdown" and co-morbidity with another anxiety disorder had fewer "paniclike" or "emotional anger" symptoms. These findings suggest that: a) the widely used term "nervous breakdown" is a popular label for different patterns of loss of emotional control; b) the type of loss of emotional control is influenced by the associated psychiatric disorder; and c) the symptoms characteristics of the "nervous breakdown" can be useful clinical markers for associated psychiatric disorders. Future research is needed to determine whether the known Hispanic entity "ataque de nervios" is simply a popular description for different aspects of well-known psychiatric disorders, or if it reflects specific demographic, environmental, personality and/or clinical

Fast Breakdown as Coronal/Ionization Waves?

NASA Astrophysics Data System (ADS)

Krehbiel, P. R.; Petersen, D.; da Silva, C. L.

2017-12-01

Studies of high-power narrow bipolar events (NBEs) have shown they are produced by a newly-recognized breakdown process called fast positive breakdown (FPB, Rison et al., 2016, doi:10.1038/ncomms10721). The breakdown was inferred to be produced by a system of positive streamers that propagate at high speed ( ˜3-6 x 107 m/s) due to occurring in a localized region of strong electric field. The polarity of the breakdown was determined from broadband interferometer (INTF) observations of the propagation direction of its VHF radiation, which was downward into the main negative charge region of a normally-electrified storm. Subsequent INTF observations being conducted in at Kennedy Space Center in Florida have shown a much greater incidence of NBEs than in New Mexico. Among the larger dataset have been clear-cut instances of some NBEs being produced by upward breakdown that would be of negative polarity. The speed and behavior of the negative breakdown is the same as that of the fast positive, leading to it being termed fast negative breakdown (FNB). The similarity (not too mention its occurrence) is surprising, given the fact that negative streamers and breakdown develops much differently than that of positive breakdown. The question is how this happens. In this study, we compare fast breakdown characteristics to well-known streamer properties as inferred from laboratory experiments and theoretical analysis. Additionally, we begin to explore the possibility that both polarities of fast breakdown are produced by what may be called coronal or ionization waves, in which the enhanced electric field produced by streamer or coronal breakdown of either polarity propagates away from the advancing front at the speed of light into a medium that is in a metastable condition of being at the threshold of hydrometeor-mediated corona onset or other ionization processes. The wave would develop at a faster speed than the streamer breakdown that gives rise to it, and thus would be

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

The structure of common-envelope remnants

NASA Astrophysics Data System (ADS)

Hall, Philip D.

2015-05-01

We investigate the structure and evolution of the remnants of common-envelope evolution in binary star systems. In a common-envelope phase, two stars become engulfed in a gaseous envelope and, under the influence of drag forces, spiral to smaller separations. They may merge to form a single star or the envelope may be ejected to leave the stars in a shorter period orbit. This process explains the short orbital periods of many observed binary systems, such as cataclysmic variables and low-mass X-ray binary systems. Despite the importance of these systems, and of common-envelope evolution to their formation, it remains poorly understood. Specifically, we are unable to confidently predict the outcome of a common-envelope phase from the properties at its onset. After presenting a review of work on stellar evolution, binary systems, common-envelope evolution and the computer programs used, we describe the results of three computational projects on common-envelope evolution. Our work specifically relates to the methods and prescriptions which are used for predicting the outcome. We use the Cambridge stellar-evolution code STARS to produce detailed models of the structure and evolution of remnants of common-envelope evolution. We compare different assumptions about the uncertain end-of-common envelope structure and envelope mass of remnants which successfully eject their common envelopes. In the first project, we use detailed remnant models to investigate whether planetary nebulae are predicted after common-envelope phases initiated by low-mass red giants. We focus on the requirement that a remnant evolves rapidly enough to photoionize the nebula and compare the predictions for different ideas about the structure at the end of a common-envelope phase. We find that planetary nebulae are possible for some prescriptions for the end-of-common envelope structure. In our second contribution, we compute a large set of single-star models and fit new formulae to the core radii of

Envelopes in eclipsing binary stars

NASA Technical Reports Server (NTRS)

Huang, S.

1972-01-01

Theoretical research on eclipsing binaries is presented. The specific areas of investigation are the following: (1) the relevance of envelopes to the study of the light curves of eclipsing binaries, (2) the disk envelope, and (3) the spherical envelope.

Molecular Characterization of the SUMO-1 Modification of RanGAP1 and Its Role in Nuclear Envelope Association

PubMed Central

Mahajan, Rohit; Gerace, Larry; Melchior, Frauke

1998-01-01

The mammalian guanosine triphosphate (GTP)ase-activating protein RanGAP1 is the first example of a protein covalently linked to the ubiquitin-related protein SUMO-1. Here we used peptide mapping, mass spectroscopy analysis, and mutagenesis to identify the nature of the link between RanGAP1 and SUMO-1. SUMO-1 is linked to RanGAP1 via glycine 97, indicating that the last 4 amino acids of this 101– amino acid protein are proteolytically removed before its attachment to RanGAP1. Recombinant SUMO-1 lacking the last four amino acids is efficiently used for modification of RanGAP1 in vitro and of multiple unknown proteins in vivo. In contrast to most ubiquitinated proteins, only a single lysine residue (K526) in RanGAP1 can serve as the acceptor site for modification by SUMO-1. Modification of RanGAP1 with SUMO-1 leads to association of RanGAP1 with the nuclear envelope (NE), where it was previously shown to be required for nuclear protein import. Sufficient information for modification and targeting resides in a 25-kD domain of RanGAP1. RanGAP1–SUMO-1 remains stably associated with the NE during many cycles of in vitro import. This indicates that removal of RanGAP1 from the NE is not a required element of nuclear protein import and suggests that the reversible modification of RanGAP1 may have a regulatory role. PMID:9442102

Internal structure of a vortex breakdown

NASA Technical Reports Server (NTRS)

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

1986-01-01

An axisymmetric vortex breakdown was well simulated by the vortex filament method. The agreement with the experiment was qualitatively good. In particular, the structure in the interior of the vortex breakdown was ensured to a great degree by the present simulation. The second breakdown, or spiral type, which occurs downstream of the first axisymmetric breakdown, was simulated more similarly to the experiment than before. It shows a kink of the vortex filaments and strong three-dimensionality. Furthermore, a relatively low velocity region was observed near the second breakdown. It was also found that it takes some time for this physical phenomenon to attain its final stage. The comparison with the experiment is getting better as time goes on. In this paper, emphasis is placed on the comparison of the simulated results with the experiment. The present results help to make clear the mechanism of a vortex breakdown.

Protoparvovirus Knocking at the Nuclear Door.

PubMed

Mäntylä, Elina; Kann, Michael; Vihinen-Ranta, Maija

2017-10-02

Protoparvoviruses target the nucleus due to their dependence on the cellular reproduction machinery during the replication and expression of their single-stranded DNA genome. In recent years, our understanding of the multistep process of the capsid nuclear import has improved, and led to the discovery of unique viral nuclear entry strategies. Preceded by endosomal transport, endosomal escape and microtubule-mediated movement to the vicinity of the nuclear envelope, the protoparvoviruses interact with the nuclear pore complexes. The capsids are transported actively across the nuclear pore complexes using nuclear import receptors. The nuclear import is sometimes accompanied by structural changes in the nuclear envelope, and is completed by intranuclear disassembly of capsids and chromatinization of the viral genome. This review discusses the nuclear import strategies of protoparvoviruses and describes its dynamics comprising active and passive movement, and directed and diffusive motion of capsids in the molecularly crowded environment of the cell.

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.

Zeeman perturbed nuclear quadrupole spin echo envelope modulations for spin 3/2 nuclei in polycrystalline specimens

NASA Astrophysics Data System (ADS)

Ramachandran, R.; Narasimhan, P. T.

The results of theoretical and experimental studies of Zeeman-perturbed nuclear quadrupole spin echo envelope modulations (ZSEEM) for spin 3/2 nuclei in polycrystalline specimens are presented. The response of the Zeeman-perturbed spin ensemble to resonant two pulse excitations has been calculated using the density matrix formalism. The theoretical calculation assumes a parallel orientation of the external r.f. and static Zeeman fields and an arbitrary orientation of these fields to the principal axes system of the electric field gradient. A numerical powder averaging procedure has been adopted to simulate the response of the polycrystalline specimens. Using a coherent pulsed nuclear quadrupole resonance spectrometer the ZSEEM patterns of the 35Cl nuclei have been recorded in polycrystalline specimens of potassium chlorate, barium chlorate, mercuric chloride (two sites) and antimony trichloride (two sites) using the π/2-τ-π/2 sequence. The theoretical and experimental ZSEEM patterns have been compared. In the case of mercuric chloride, the experimental 35Cl ZSEEM patterns are found to be nearly identical for the two sites and correspond to a near-zero value of the asymmetry parameter, η, of the electric field gradient tensor. The difference in the η values for the two 35Cl sites (η ˜0·06 and η˜0·16) in antimony trichloride is clearly reflected in the experimental and theoretical ZSEEM patterns. The present study indicates the feasibility of evaluating η for spin 3/2 nuclei in polycrystalline specimens from ZSEEM investigations.

Nuclear envelope-distributed CD147 interacts with and inhibits the transcriptional function of RING1 and promotes melanoma cell motility.

PubMed

Chen, Junchen; Peng, Cong; Lei, Li; Zhang, Jianglin; Zeng, Weiqi; Chen, Xiang

2017-01-01

Melanoma accounts for nearly 80% of all deaths associated with skin cancer.CD147 plays a very important role in melanoma progression and the expression level may correlate with tumor malignancy. RING1 can bind DNA and act as a transcriptional repressor, play an important role in the aggressive phenotype in melanoma. The interactions between CD147 and RING1 were identified with a yeast two-hybrid and RING1 interacted with CD147 through the transmembrane domain. RING1 inhibits CD147's capability promoting melanoma cell migration. In conclusion, the study identified novel interactions between CD147 and RING1, recovered CD147 nuclear envelope distribution in melanoma cells, and suggested a new mechanism underlying how cytoplasmic CD147 promotes melanoma development.

Nuclear envelope-distributed CD147 interacts with and inhibits the transcriptional function of RING1 and promotes melanoma cell motility

PubMed Central

Peng, Cong; Lei, Li; Zhang, Jianglin; Zeng, Weiqi; Chen, Xiang

2017-01-01

Melanoma accounts for nearly 80% of all deaths associated with skin cancer.CD147 plays a very important role in melanoma progression and the expression level may correlate with tumor malignancy. RING1 can bind DNA and act as a transcriptional repressor, play an important role in the aggressive phenotype in melanoma. The interactions between CD147 and RING1 were identified with a yeast two-hybrid and RING1 interacted with CD147 through the transmembrane domain. RING1 inhibits CD147’s capability promoting melanoma cell migration. In conclusion, the study identified novel interactions between CD147 and RING1, recovered CD147 nuclear envelope distribution in melanoma cells, and suggested a new mechanism underlying how cytoplasmic CD147 promotes melanoma development. PMID:28832687

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Young, Kevin G.; University of Ottawa Center for Neuromuscular Disease, University of Ottawa, Ottawa, Ontario; 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 precedingmore » 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.« less

Breach of the nuclear lamina during assembly of herpes simplex viruses.

PubMed

Morrison, Lynda A; DeLassus, Gregory S

2011-01-01

Beneath the inner nuclear membrane lies the dense meshwork of the nuclear lamina, which provides structural support for the nuclear envelope and serves as an important organizing center for a number of nuclear and cytoplasmic constituents and processes. Herpesviruses have a significant and wide-ranging impact on human health, and their capacity to replicate and cause disease includes events that occur in the host cell nucleus. Herpesviruses begin assembly of progeny virus in the nuclei of infected cells and their capsids must escape the confines of the nucleus by budding through the inner nuclear membrane (INM) to proceed with later stages of virion assembly and egress. Access of viral capsids to the INM thus necessitates disruption of the dense nuclear lamina layer. We review herpesvirus effects on the nuclear lamina and in particular the roles of the herpes simplex virus-encoded nuclear envelope complex and viral kinases on lamin phosphorylation, dissociation, and nucleocapsid envelopment at the INM.

Geriatric Sexuality Breakdown Syndrome.

ERIC Educational Resources Information Center

Kaas, Merrie Jean

1981-01-01

Focuses on the relationship between social environment and the older individual. By utilizing the Social Breakdown Syndrome a cycle of events is defined by the Geriatric Sexuality Breakdown Syndrome, in which an older individual is initially predisposed to diminished sexual activity to the end point of self-identification as nonsexual. (Author)

Evaluation of eco-friendly zwitterionic detergents for enveloped virus inactivation.

PubMed

Conley, Lynn; Tao, Yinying; Henry, Alexis; Koepf, Edward; Cecchini, Douglas; Pieracci, John; Ghose, Sanchayita

2017-04-01

Inclusion of a detergent in protein biotherapeutic purification processes is a simple and very robust method for inactivating enveloped viruses. The detergent Triton X-100 has been used for many years and is part of the production process of several commercial therapeutic proteins. However, recent ecological studies have suggested that Triton X-100 and its break-down products can potentially behave as endocrine disrupters in aquatic organisms, raising concerns from an environmental impact perspective. As such, discharge of Triton X-100 into the waste water treatment plants is regulated in some jurisdictions, and alternative detergents for viral inactivation are required. In this work, we report on the identification and evaluation of more eco-friendly detergents as viable replacements for Triton X-100. Five detergent candidates with low to moderate environmental impact were initially identified and evaluated with respect to protein stability, followed by proof-of-concept virus inactivation studies using a model enveloped virus. From the set of candidates lauryldimethylamine N-oxide (LDAO) was identified as the most promising detergent due to its low ecotoxicity, robust anti-viral activity (LRV >4 at validation set-point conditions with X-MuLX), and absence of any negative impact on protein function. This detergent exhibited effective and robust virus inactivation in a broad range of protein concentrations, solution conductivities, pHs, and in several different cell culture fluid matrices. The only process parameter which correlated with reduced virus inactivation potency was LDAO concentration, and then only when the concentration was reduced to below the detergent's critical micelle concentration (CMC). Additionally, this work also demonstrated that LDAO was cleared to below detectable levels after Protein A affinity chromatography, making it suitable for use in a platform process that utilizes this chromatographic mode for protein capture. All these findings

Protein targeting and integration signal for the chloroplastic outer envelope membrane.

PubMed Central

Li, H M; Chen, L J

1996-01-01

Most proteins in chloroplasts are encoded by the nuclear genome and synthesized in the cytosol. With the exception of most quter envelope membrane proteins, nuclear-encoded chloroplastic proteins are synthesized with N-terminal extensions that contain the chloroplast targeting information of these proteins. Most outer membrane proteins, however, are synthesized without extensions in the cytosol. Therefore, it is not clear where the chloroplastic outer membrane targeting information resides within these polypeptides. We have analyzed a chloroplastic outer membrane protein, OEP14 (outer envelope membrane protein of 14 kD, previously named OM14), and localized its outer membrane targeting and integration signal to the first 30 amino acids of the protein. This signal consists of a positively charged N-terminal portion followed by a hydrophobic core, bearing resemblance to the signal peptides of proteins targeted to the endoplasmic reticulum. However, a chimeric protein containing this signal fused to a passenger protein did not integrate into the endoplasmic reticulum membrane. Furthermore, membrane topology analysis indicated that the signal inserts into the chloroplastic outer membrane in an orientation opposite to that predicted by the "positive inside" rule. PMID:8953775

LEM4/ANKLE-2 deficiency impairs post-mitotic re-localization of BAF, LAP2α and LaminA to the nucleus, causes nuclear envelope instability in telophase and leads to hyperploidy in HeLa cells.

PubMed

Snyers, Luc; Erhart, Renate; Laffer, Sylvia; Pusch, Oliver; Weipoltshammer, Klara; Schöfer, Christian

2018-01-01

The human LEM-domain protein family is involved in fundamental aspects of nuclear biology. The LEM-domain interacts with the barrier-to-autointegration factor (BAF), which itself binds DNA. LEM-domain proteins LAP2, emerin and MAN1 are proteins of the inner nuclear membrane; they have important functions: maintaining the integrity of the nuclear lamina and regulating gene expression at the nuclear periphery. LEM4/ANKLE-2 has been proposed to participate in nuclear envelope reassembly after mitosis and to mediate dephosphorylation of BAF through binding to phosphatase PP2A. Here, we used CRISPR/Cas9 to create several cell lines deficient in LEM4/ANKLE-2. By using time-lapse video microscopy, we show that absence of this protein severely compromises the post mitotic re-association of the nuclear proteins BAF, LAP2α and LaminA to chromosomes. These defects give rise to a strong mechanical instability of the nuclear envelope in telophase and to a chromosomal instability leading to increased number of hyperploid cells. Reintroducing LEM4/ANKLE-2 in the cells by transfection could efficiently restore the telophase association of BAF and LAP2α to the chromosomes. This rescue phenotype was abolished for N- or C-terminally truncated mutants that had lost the capacity to bind PP2A. We demonstrate also that, in addition to binding to PP2A, LEM4/ANKLE-2 binds BAF through its LEM-domain, providing further evidence for a generic function of this domain as a principal interactor of BAF. Copyright © 2017 Elsevier GmbH. All rights reserved.

Skin breakdown in acute care pediatrics.

PubMed

Suddaby, Elizabeth C; Barnett, Scott D; Facteau, Lorna

2006-04-01

The purpose of this study was to develop a simple, single-page measurement tool that evaluates risk of skin breakdown in the peadiatric population and apply it to the acutely hospitalized child. Data were collected over a 15-month period from 347 patients on four in-patient units (PICU, medical-surgical, oncology, and adolescents) on skin breakdown using the AHCPR staging guidelines and compared to the total score on the Starkid SkinScale in order to determine its ability to predict skin breakdown. The inter-rater reliability of the Starkid Skin Scale was r2 = 0.85 with an internal reliablity of 0.71. The sensitivity of the total score was low (17.5%) but highly specific (98.5%). The prevalence of skin breakdown in the acutely hospitalized child was 23%, the majority (77.5%) occurring as erythema of the skin. Buttocks, perineum, and occiput were the most common locations of breakdown. Occiput breakdown was more common in critically ill (PICU) patients while diaper dermatitis was more common in the general medical-surgical population.

Cortical processing of dynamic sound envelope transitions.

PubMed

Zhou, Yi; Wang, Xiaoqin

2010-12-08

Slow envelope fluctuations in the range of 2-20 Hz provide important segmental cues for processing communication sounds. For a successful segmentation, a neural processor must capture envelope features associated with the rise and fall of signal energy, a process that is often challenged by the interference of background noise. This study investigated the neural representations of slowly varying envelopes in quiet and in background noise in the primary auditory cortex (A1) of awake marmoset monkeys. We characterized envelope features based on the local average and rate of change of sound level in envelope waveforms and identified envelope features to which neurons were selective by reverse correlation. Our results showed that envelope feature selectivity of A1 neurons was correlated with the degree of nonmonotonicity in their static rate-level functions. Nonmonotonic neurons exhibited greater feature selectivity than monotonic neurons in quiet and in background noise. The diverse envelope feature selectivity decreased spike-timing correlation among A1 neurons in response to the same envelope waveforms. As a result, the variability, but not the average, of the ensemble responses of A1 neurons represented more faithfully the dynamic transitions in low-frequency sound envelopes both in quiet and in background noise.

Role of Nuclear Lamina in Gene Repression and Maintenance of Chromosome Architecture in the Nucleus.

PubMed

Shevelyov, Y Y; Ulianov, S V

2018-04-01

Nuclear lamina is a protein meshwork composed of lamins and lamin-associated proteins that lines the nuclear envelope from the inside and forms repressive transcription compartment. The review presents current data on the contribution of nuclear lamina to the repression of genes located in this compartment and on the mechanisms of chromatin attachment to the nuclear envelope.

Nonstationary envelope process and first excursion probability

NASA Technical Reports Server (NTRS)

Yang, J.

1972-01-01

A definition of the envelope of nonstationary random processes is proposed. The establishment of the envelope definition makes it possible to simulate the nonstationary random envelope directly. Envelope statistics, such as the density function, joint density function, moment function, and level crossing rate, which are relevent to analyses of catastrophic failure, fatigue, and crack propagation in structures, are derived. Applications of the envelope statistics to the prediction of structural reliability under random loadings are discussed in detail.

Cyclooxygenase-2 regulated by the nuclear factor-κB pathway plays an important role in endometrial breakdown in a female mouse menstrual-like model.

PubMed

Xu, Xiangbo; Chen, Xihua; Li, Yunfeng; Cao, Huizi; Shi, Cuige; Guan, Shuo; Zhang, Shucheng; He, Bin; Wang, Jiedong

2013-08-01

The role of prostaglandins (PGs) in menstruation has long been proposed. Although evidence from studies on human and nonhuman primates supports the involvement of PGs in menstruation, whether PGs play an obligatory role in the process remains unclear. Although cyclooxygenase (COX) inhibitors have been used in the treatment of irregular uterine bleeding, the mechanism involved has not been elucidated. In this study, we used a recently established mouse menstrual-like model for investigating the role of COX in endometrial breakdown and its regulation. Administration of the nonspecific COX inhibitor indomethacin and the COX-2 selective inhibitor DuP-697 led to inhibition of the menstrual-like process. Furthermore, immunostaining analysis showed that the nuclear factor (NF)κB proteins P50, P65, and COX-2 colocalized in the outer decidual stroma at 12 to 16 hours after progesterone withdrawal. Chromatin immunoprecipitation analysis showed that NFκB binding to the Cox-2 promoter increased at 12 hours after progesterone withdrawal in vivo, and real-time PCR analysis showed that the NFκB inhibitors pyrrolidine dithiocarbamate and MG-132 inhibited Cox-2 mRNA expression in vivo and in vitro, respectively. Furthermore, COX-2 and NFκB inhibitors similarly reduced endometrial breakdown, suggesting that NFκB/COX-2-derived PGs play a critical role in this process. In addition, the CD45(+) leukocyte numbers were sharply reduced following indomethacin (COX-1 and COX-2 inhibitor), DuP-697 (COX-2 inhibitor), and pyrrolidine dithiocarbamate (NFκB inhibitor) treatment. Collectively, these data indicate that NFκB/COX-2-induced PGs regulate leukocyte influx, leading to endometrial breakdown.

Laser-induced breakdown spectroscopy of light water reactor simulated used nuclear fuel: Main oxide phase

DOE PAGES

Campbell, Keri R.; Judge, Elizabeth J.; Barefield, James E.; ...

2017-04-22

We show the analysis of light water reactor simulated used nuclear fuel using laser-induced breakdown spectroscopy (LIBS) is explored using a simplified version of the main oxide phase. The main oxide phase consists of the actinides, lanthanides, and zirconium. The purpose of this study is to develop a rapid, quantitative technique for measuring zirconium in a uranium dioxide matrix without the need to dissolve the material. A second set of materials including cerium oxide is also analyzed to determine precision and limit of detection (LOD) using LIBS in a complex matrix. Two types of samples are used in this study:more » binary and ternary oxide pellets. The ternary oxide, (U,Zr,Ce)O 2 pellets used in this study are a simplified version the main oxide phase of used nuclear fuel. The binary oxides, (U,Ce)O 2 and (U,Zr)O 2 are also examined to determine spectral emission lines for Ce and Zr, potential spectral interferences with uranium and baseline LOD values for Ce and Zr in a UO 2 matrix. In the spectral range of 200 to 800 nm, 33 cerium lines and 25 zirconium lines were identified and shown to have linear correlation values (R 2) > 0.97 for both the binary and ternary oxides. The cerium LOD in the (U,Ce)O 2 matrix ranged from 0.34 to 1.08 wt% and 0.94 to 1.22 wt% in (U,Ce,Zr)O 2 for 33 of Ce emission lines. The zirconium limit of detection in the (U,Zr)O 2 matrix ranged from 0.84 to 1.15 wt% and 0.99 to 1.10 wt% in (U,Ce,Zr)O 2 for 25 Zr lines. Finally, the effect of multiple elements in the plasma and the impact on the LOD is discussed.« less

Nuclear γ-tubulin associates with nucleoli and interacts with tumor suppressor protein C53.

PubMed

Hořejší, Barbora; Vinopal, Stanislav; Sládková, Vladimíra; Dráberová, Eduarda; Sulimenko, Vadym; Sulimenko, Tetyana; Vosecká, Věra; Philimonenko, Anatoly; Hozák, Pavel; Katsetos, Christos D; Dráber, Pavel

2012-01-01

γ-Tubulin is assumed to be a typical cytosolic protein necessary for nucleation of microtubules from microtubule organizing centers. Using immunolocalization and cell fractionation techniques in combination with siRNAi and expression of FLAG-tagged constructs, we have obtained evidence that γ-tubulin is also present in nucleoli of mammalian interphase cells of diverse cellular origins. Immunoelectron microscopy has revealed γ-tubulin localization outside fibrillar centers where transcription of ribosomal DNA takes place. γ-Tubulin was associated with nucleolar remnants after nuclear envelope breakdown and could be translocated to nucleoli during mitosis. Pretreatment of cells with leptomycin B did not affect the distribution of nuclear γ-tubulin, making it unlikely that rapid active transport via nuclear pores participates in the transport of γ-tubulin into the nucleus. This finding was confirmed by heterokaryon assay and time-lapse imaging of photoconvertible protein Dendra2 tagged to γ-tubulin. Immunoprecipitation from nuclear extracts combined with mass spectrometry revealed an association of γ-tubulin with tumor suppressor protein C53 located at multiple subcellular compartments including nucleoli. The notion of an interaction between γ-tubulin and C53 was corroborated by pull-down and co-immunoprecipitation experiments. Overexpression of γ-tubulin antagonized the inhibitory effect of C53 on DNA damage G(2) /M checkpoint activation. The combined results indicate that aside from its known role in microtubule nucleation, γ-tubulin may also have nuclear-specific function(s). Copyright © 2011 Wiley Periodicals, Inc.

Strategic disruption of nuclear pores structure, integrity and barrier for nuclear apoptosis.

PubMed

Shahin, Victor

2017-08-01

Apoptosis is a programmed cell death playing key roles in physiology and pathophysiology of multi cellular organisms. Its nuclear manifestation requires transmission of the death signals across the nuclear pore complexes (NPCs). In strategic sequential steps apoptotic factors disrupt NPCs structure, integrity and barrier ultimately leading to nuclear breakdown. The present review reflects on these steps. Copyright © 2017 Elsevier Ltd. All rights reserved.

Vesicular PtdIns(3,4,5)P3 and Rab7 are key effectors of sea urchin zygote nuclear membrane fusion.

PubMed

Lete, Marta G; Byrne, Richard D; Alonso, Alicia; Poccia, Dominic; Larijani, Banafshé

2017-01-15

Regulation of nuclear envelope dynamics is an important example of the universal phenomena of membrane fusion. The signalling molecules involved in nuclear membrane fusion might also be conserved during the formation of both pronuclear and zygote nuclear envelopes in the fertilised egg. Here, we determine that class-I phosphoinositide 3-kinases (PI3Ks) are needed for in vitro nuclear envelope formation. We show that, in vivo, PtdIns(3,4,5)P 3 is transiently located in vesicles around the male pronucleus at the time of nuclear envelope formation, and around male and female pronuclei before membrane fusion. We illustrate that class-I PI3K activity is also necessary for fusion of the female and male pronuclear membranes. We demonstrate, using coincidence amplified Förster resonance energy transfer (FRET) monitored using fluorescence lifetime imaging microscopy (FLIM), a protein-lipid interaction of Rab7 GTPase and PtdIns(3,4,5)P 3 that occurs during pronuclear membrane fusion to create the zygote nuclear envelope. We present a working model, which includes several molecular steps in the pathways controlling fusion of nuclear envelope membranes. © 2017. Published by The Company of Biologists Ltd.

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.

Investigation of Dielectric Breakdown Characteristics for Double-break Vacuum Interrupter and Dielectric Breakdown Probability Distribution in Vacuum Interrupter

NASA Astrophysics Data System (ADS)

Shioiri, Tetsu; Asari, Naoki; Sato, Junichi; Sasage, Kosuke; Yokokura, Kunio; Homma, Mitsutaka; Suzuki, Katsumi

To investigate the reliability of equipment of vacuum insulation, a study was carried out to clarify breakdown probability distributions in vacuum gap. Further, a double-break vacuum circuit breaker was investigated for breakdown probability distribution. The test results show that the breakdown probability distribution of the vacuum gap can be represented by a Weibull distribution using a location parameter, which shows the voltage that permits a zero breakdown probability. The location parameter obtained from Weibull plot depends on electrode area. The shape parameter obtained from Weibull plot of vacuum gap was 10∼14, and is constant irrespective non-uniform field factor. The breakdown probability distribution after no-load switching can be represented by Weibull distribution using a location parameter. The shape parameter after no-load switching was 6∼8.5, and is constant, irrespective of gap length. This indicates that the scatter of breakdown voltage was increased by no-load switching. If the vacuum circuit breaker uses a double break, breakdown probability at low voltage becomes lower than single-break probability. Although potential distribution is a concern in the double-break vacuum cuicuit breaker, its insulation reliability is better than that of the single-break vacuum interrupter even if the bias of the vacuum interrupter's sharing voltage is taken into account.

Modular shipbuilding and its relevance to construction of nuclear power plants. Master's thesis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Seubert, T.W.

1988-05-01

The modern techniques of modular shipbuilding based on the Product Work Breakdown Structure as developed at the Ishikawajima-Harima Heavy Industries Co., Ltd. of Japan are examined and compared to conventional shipbuilding methods. The application of the Product Work Breakdown Structure in the building of the U.S. Navy's DDG-51 class ship at Bath Iron Works is described and compared to Japanese shipbuilding practices. Implementation of the Product Work Breakdown Structure at Avondale Shipyards, Incorporated is discussed and compared to Bath Iron Works shipbuilding practices. A proposed generic implementation of the Product Work Breakdown Structure to the modular construction of nuclear powermore » plants is described. Specific conclusions for the application of Product Work Breakdown Structure to the construction of a light water reactor nuclear power plant are discussed.« less

ROS are critical for endometrial breakdown via NF-κB-COX-2 signaling in a female mouse menstrual-like model.

PubMed

Wu, Bin; Chen, Xihua; He, Bin; Liu, Shuyan; Li, Yunfeng; Wang, Qianxing; Gao, Haijun; Wang, Shufang; Liu, Jianbing; Zhang, Shucheng; Xu, Xiangbo; Wang, Jiedong

2014-09-01

Progesterone withdrawal triggers endometrial breakdown and shedding during menstruation. Menstruation results from inflammatory responses; however, the role of reactive oxygen species (ROS) in menstruation remains unclear. In this study, we explored the role of ROS in endometrial breakdown and shedding. We found that ROS levels were significantly increased before endometrial breakdown in a mouse menstrual-like model. Vaginal smear inspection, morphology of uterine horns, and endometrial histology examination showed that a broad range of ROS scavengers significantly inhibited endometrial breakdown in this model. Furthermore, Western blot and immunohistochemical analysis showed that the intracellular translocation of p50 and p65 from the cytoplasm into the nucleus was blocked by ROS scavengers and real-time PCR showed that cyclooxygenase-2 (COX-2) mRNA expression was decreased by ROS scavengers. Similar changes also occurred in human stromal cells in vitro. Furthermore, Western blotting and real-time PCR showed that one ROS, hydrogen peroxide (H2O2), promoted translocation of p50 and p65 from the cytoplasm to the nucleus and increased COX-2 mRNA expression along with progesterone maintenance. The nuclear factor κB inhibitor MG132 reduced the occurrence of these changes in human stromal cells in vitro. Viewed as a whole, our results provide evidence that certain ROS are important for endometrial breakdown and shedding in a mouse menstrual-like model and function at least partially via nuclear factor-κB/COX-2 signaling. Similar changes observed in human stromal cells could also implicate ROS as important mediators of human menstruation.

Acute Depletion of Diacylglycerol from the Cis-Golgi Affects Localised Nuclear Envelope Morphology During Mitosis.

PubMed

Chung, Gary Hong Chun; Domart, Marie Charlotte; Peddie, Christopher; Mantell, Judith; Mclaverty, Kieran; Arabiotorre, Angela; Hodgson, Lorna; Byrne, Richard D; Verkade, Paul; Arkill, Kenton; Collinson, Lucy M; Larijani, Banafshe

2018-06-12

Dysregulation of nuclear envelope (NE) assembly results in various cancers; for example, renal and some lung carcinomas ensue due to NE malformation. The NE is a dynamic membrane compartment and its completion during mitosis, is a highly regulated process but the detailed mechanism still remains incompletely understood. Previous studies have found isolated diacylglycerol (DAG) containing vesicles are essential for completing the fusion of NE in non-somatic cells. We investigated the impact of DAG depletion from cis-Golgi in mammalian cells on NE reassembly. Using advanced electron microscopy, we observed an enriched DAG population of vesicles at the vicinity of the NE gaps of telophase mammalian cells. We applied a miniSOG-C1-domain tag that localized DAG-enriched vesicles at the perinuclear region, which suggested the existence of NE fusogenic vesicles. We quantified the impact of Golgi-DAG depletion by measuring the in situ NE rim curvature of the re-forming NE. The rim curvature in these cells was significantly reduced compared with controls, which indicated a localized defect in NE morphology. Our novel results demonstrate the significance of the role of DAG from cis-Golgi for the regulation of NE assembly. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.

Sequence and characterization of cytoplasmic nuclear protein import factor p97

PubMed Central

1995-01-01

Nuclear location sequence-mediated binding of karyophilic proteins to the nuclear pore complexes is one of the earliest steps in nuclear protein import. We previously identified two cytosolic proteins that reconstitute this step in a permeabilized cell assay: the 54/56-kD NLS receptor and p97. A monoclonal antibody to p97 localizes the protein to the cytoplasm and the nuclear envelope. p97 is extracted from nuclear envelopes under the same conditions as the O-glycosylated nucleoporins indicating a tight association with the pore complex. The antibody inhibits import in a permeabilized cell assay but does not affect binding of karyophiles to the nuclear pore complex. Immunodepletion of p97 renders the cytosol inactive for import and identifies at least three other cytosolic proteins that interact with p97. cDNA cloning of p97 shows that it is a unique protein containing 23 cysteine residues. Recombinant p97 binds zinc and a bound metal ion is required for the nuclear envelope binding activity of the protein. PMID:7615630

ELECTRON MICROSCOPY OF MITOSIS IN A RADIOSENSITIVE GIANT AMOEBA

PubMed Central

Daniels, E. W.; Roth, L. E.

1964-01-01

Various aspects of the ultrastructure of the dividing nuclei in the large radiosensitive amoeba Pelomyxa illinoisensis are demonstrated. Evidence of nuclear envelope breakdown is presented, and membrane fragments are traced throughout metaphase to envelope reconstruction in anaphase and telophase. Annuli in the nuclear envelope and its fragments are shown throughout mitosis. During metaphase and anaphase some 15 to 20 mitochondria are aligned at each end of the spindle, and are called polar mitochondria. The radioresistant amoebae Pelomyxa carolinensis and Amoeba proteus do not have polar mitochondria, and Pelomyxa illinoisensis is unique in this regard. The shape of the P. illinoisensis interphase nucleoli differs from that in the two radioresistant species, and certain aspects of nucleolar dissolution in the prophase vary. Helical coils in the interphase nucleoplasm are similar to those in the radioresistant amoebae. A "blister" phase in the flatly shaped telophase nuclei of P. illinoisensis is described which is interpreted to be the result of a rapid nuclear expansion leading to the formation of the normal spherical interphase nuclei. PMID:14105218

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kind, Barbara, E-mail: barbara.kind@uniklinikum-dresden.de; Koehler, Katrin, E-mail: katrin.koehler@uniklinikum-dresden.de; Lorenz, Mike, E-mail: mlorenz@mpi-cbg.de

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 andmore » 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.« less

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.

STS-8 postal Stamp envelope

NASA Technical Reports Server (NTRS)

1983-01-01

STS-8 postal Stamp envelope with Challenger insignia, USA eagle stamp, 25th NASA anniversary stamp. The envelope is stamped with various postmarks, one saying Kennedy Space Center, Fl., another saying 'Returned to earth, Edwards AFB, CA'.

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.

Humidity effects on wire insulation breakdown strength.

DOE Office of Scientific and Technical Information (OSTI.GOV)

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 layermore » 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.« less

The type 2 dengue virus envelope protein interacts with small ubiquitin-like modifier-1 (SUMO-1) conjugating enzyme 9 (Ubc9).

PubMed

Chiu, Mei-Wui; Shih, Hsiu-Ming; Yang, Tsung-Han; Yang, Yun-Liang

2007-05-01

Dengue viruses are mosquito-borne flaviviruses and may cause the life-threatening dengue hemorrhagic fever and dengue shock syndrome. Its envelope protein is responsible mainly for the virus attachment and entry to host cells. To identify the human cellular proteins interacting with the envelope protein of dengue virus serotype 2 inside host cells, we have performed a screening with the yeast-two-hybrid-based "Functional Yeast Array". Interestingly, the small ubiquitin-like modifier-1 conjugating enzyme 9 protein, modulating cellular processes such as those regulating signal transduction and cell growth, was one of the candidates interacting with the dengue virus envelope protein. With co-precipitation assay, we have demonstrated that it indeed could interact directly with the Ubc9 protein. Site-directed mutagenesis has demonstrated that Ubc9 might interact with the E protein via amino acid residues K51 and K241. Furthermore, immunofluorescence microscopy has shown that the DV2E-EGFP proteins tended to progress toward the nuclear membrane and co-localized with Flag-Ubc9 proteins around the nuclear membrane in the cytoplasmic side, and DV2E-EGFP also shifted the distribution of Flag-Ubc9 from evenly in the nucleus toward concentrating around the nuclear membrane in the nucleic side. In addition, over-expression of Ubc9 could reduce the plaque formation of the dengue virus in mammalian cells. This is the first report that DV envelope proteins can interact with the protein of sumoylation system and Ubc9 may involve in the host defense system to prevent virus propagation.

Caenorhabditis elegans polo-like kinase PLK-1 is required for merging parental genomes into a single nucleus.

PubMed

Rahman, Mohammad M; Munzig, Mandy; Kaneshiro, Kiyomi; Lee, Brandon; Strome, Susan; Müller-Reichert, Thomas; Cohen-Fix, Orna

2015-12-15

Before the first zygotic division, the nuclear envelopes of the maternal and paternal pronuclei disassemble, allowing both sets of chromosomes to be incorporated into a single nucleus in daughter cells after mitosis. We found that in Caenorhabditis elegans, partial inactivation of the polo-like kinase PLK-1 causes the formation of two nuclei, containing either the maternal or paternal chromosomes, in each daughter cell. These two nuclei gave rise to paired nuclei in all subsequent cell divisions. The paired-nuclei phenotype was caused by a defect in forming a gap in the nuclear envelopes at the interface between the two pronuclei during the first mitotic division. This was accompanied by defects in chromosome congression and alignment of the maternal and paternal metaphase plates relative to each other. Perturbing chromosome congression by other means also resulted in failure to disassemble the nuclear envelope between the two pronuclei. Our data further show that PLK-1 is needed for nuclear envelope breakdown during early embryogenesis. We propose that during the first zygotic division, PLK-1-dependent chromosome congression and metaphase plate alignment are necessary for the disassembly of the nuclear envelope between the two pronuclei, ultimately allowing intermingling of the maternal and paternal chromosomes. © 2015 Rahman et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

Nuclear Import of the Retrotransposon Tf1 Is Governed by a Nuclear Localization Signal That Possesses a Unique Requirement for the FXFG Nuclear Pore Factor Nup124p

PubMed Central

Dang, Van-Dinh; Levin, Henry L.

2000-01-01

Retroviruses, such as human immunodeficiency virus, that infect nondividing cells generate integration precursors that must cross the nuclear envelope to reach the host genome. As a model for retroviruses, we investigated the nuclear entry of Tf1, a long-terminal-repeat-containing retrotransposon of the fission yeast Schizosaccharomyces pombe. Because the nuclear envelope of yeasts remains intact throughout the cell cycle, components of Tf1 must be transported through the envelope before integration can occur. The nuclear localization of the Gag protein of Tf1 is different from that of other proteins tested in that it has a specific requirement for the FXFG nuclear pore factor, Nup124p. Using extensive mutagenesis, we found that Gag contained three nuclear localization signals (NLSs) which, when included individually in a heterologous protein, were sufficient to direct nuclear import. In the context of the intact transposon, mutations in the NLS that mapped to the first 10 amino acid residues of Gag significantly impaired Tf1 retrotransposition and abolished nuclear localization of Gag. Interestingly, this NLS activity in the heterologous protein was specifically dependent upon the presence of Nup124p. Deletion analysis of heterologous proteins revealed the surprising result that the residues in Gag with the NLS activity were independent from the residues that conveyed the requirement for Nup124p. In fact, a fragment of Gag that lacked NLS activity, residues 10 to 30, when fused to a heterologous protein, was sufficient to cause the classical NLS of simian virus 40 to require Nup124p for nuclear import. Within the context of the current understanding of nuclear import, these results represent the novel case of a short amino acid sequence that specifies the need for a particular nuclear pore complex protein. PMID:11003674

Nuclear import of the retrotransposon Tf1 is governed by a nuclear localization signal that possesses a unique requirement for the FXFG nuclear pore factor Nup124p.

PubMed

Dang, V D; Levin, H L

2000-10-01

Retroviruses, such as human immunodeficiency virus, that infect nondividing cells generate integration precursors that must cross the nuclear envelope to reach the host genome. As a model for retroviruses, we investigated the nuclear entry of Tf1, a long-terminal-repeat-containing retrotransposon of the fission yeast Schizosaccharomyces pombe. Because the nuclear envelope of yeasts remains intact throughout the cell cycle, components of Tf1 must be transported through the envelope before integration can occur. The nuclear localization of the Gag protein of Tf1 is different from that of other proteins tested in that it has a specific requirement for the FXFG nuclear pore factor, Nup124p. Using extensive mutagenesis, we found that Gag contained three nuclear localization signals (NLSs) which, when included individually in a heterologous protein, were sufficient to direct nuclear import. In the context of the intact transposon, mutations in the NLS that mapped to the first 10 amino acid residues of Gag significantly impaired Tf1 retrotransposition and abolished nuclear localization of Gag. Interestingly, this NLS activity in the heterologous protein was specifically dependent upon the presence of Nup124p. Deletion analysis of heterologous proteins revealed the surprising result that the residues in Gag with the NLS activity were independent from the residues that conveyed the requirement for Nup124p. In fact, a fragment of Gag that lacked NLS activity, residues 10 to 30, when fused to a heterologous protein, was sufficient to cause the classical NLS of simian virus 40 to require Nup124p for nuclear import. Within the context of the current understanding of nuclear import, these results represent the novel case of a short amino acid sequence that specifies the need for a particular nuclear pore complex protein.

Quantification of the Spatial Organization of the Nuclear Lamina as a Tool for Cell Classification

PubMed Central

Righolt, Christiaan H.; Zatreanu, Diana A.; Raz, Vered

2013-01-01

The nuclear lamina is the structural scaffold of the nuclear envelope that plays multiple regulatory roles in chromatin organization and gene expression as well as a structural role in nuclear stability. The lamina proteins, also referred to as lamins, determine nuclear lamina organization and define the nuclear shape and the structural integrity of the cell nucleus. In addition, lamins are connected with both nuclear and cytoplasmic structures forming a dynamic cellular structure whose shape changes upon external and internal signals. When bound to the nuclear lamina, the lamins are mobile, have an impact on the nuclear envelop structure, and may induce changes in their regulatory functions. Changes in the nuclear lamina shape cause changes in cellular functions. A quantitative description of these structural changes could provide an unbiased description of changes in cellular function. In this review, we describe how changes in the nuclear lamina can be measured from three-dimensional images of lamins at the nuclear envelope, and we discuss how structural changes of the nuclear lamina can be used for cell classification. PMID:27335676

Quantification of the Spatial Organization of the Nuclear Lamina as a Tool for Cell Classification.

PubMed

Righolt, Christiaan H; Zatreanu, Diana A; Raz, Vered

2013-01-01

The nuclear lamina is the structural scaffold of the nuclear envelope that plays multiple regulatory roles in chromatin organization and gene expression as well as a structural role in nuclear stability. The lamina proteins, also referred to as lamins, determine nuclear lamina organization and define the nuclear shape and the structural integrity of the cell nucleus. In addition, lamins are connected with both nuclear and cytoplasmic structures forming a dynamic cellular structure whose shape changes upon external and internal signals. When bound to the nuclear lamina, the lamins are mobile, have an impact on the nuclear envelop structure, and may induce changes in their regulatory functions. Changes in the nuclear lamina shape cause changes in cellular functions. A quantitative description of these structural changes could provide an unbiased description of changes in cellular function. In this review, we describe how changes in the nuclear lamina can be measured from three-dimensional images of lamins at the nuclear envelope, and we discuss how structural changes of the nuclear lamina can be used for cell classification.

High-voltage subnanosecond dielectric breakdown

NASA Astrophysics Data System (ADS)

Mankowski, John Jerome

Current interests in ultrawideband radar sources are in the microwave regime, which correspond to voltage pulse risetimes less than a nanosecond. Some new sources, including the Phillips Laboratory Hindenberg series of hydrogen gas switched pulsers use hydrogen at hundreds of atmospheres of pressure in the switch. Unfortunately, the published data of electrical breakdown of gas and liquid media at these time lengths are relatively scarce. A study was conducted on the electrical breakdown properties of liquid and gas dielectrics at subnanosecond and nanoseconds. Two separate voltage sources with pulse risetimes less than 400 ps were developed. Diagnostic probes were designed and tested for their capability of detecting high voltage pulses at these fast risetimes. A thorough investigation into E-field strengths of liquid and gas dielectrics at breakdown times ranging from 0.4 to 5 ns was performed. The voltage polarity dependence on breakdown strength is observed. Streak camera images of streamer formation were taken. The effect of ultraviolet radiation, incident upon the gap, on statistical lag time was determined.

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.

Breakdown of single spin-fluid model in the heavily hole-doped superconductor CsFe2As2

NASA Astrophysics Data System (ADS)

Zhao, D.; Li, S. J.; Wang, N. Z.; Li, J.; Song, D. W.; Zheng, L. X.; Nie, L. P.; Luo, X. G.; Wu, T.; Chen, X. H.

2018-01-01

Although Fe-based superconductors are correlated electronic systems with multiorbital, previous nuclear magnetic resonance (NMR) measurement suggests that a single spin-fluid model is sufficient to describe its spin behavior. Here, we first observed the breakdown of single spin-fluid model in a heavily hole-doped Fe-based superconductor CsFe2As2 by site-selective NMR measurement. At high-temperature regime, both Knight shift and nuclear spin-lattice relaxation at 133Cs and 75As nuclei exhibit distinct temperature-dependent behavior, suggesting the breakdown of the single spin-fluid model in CsFe2As2 . This is ascribed to the coexistence of both localized and itinerant spin degree of freedom at 3 d orbitals, which is consistent with the orbital-selective Mott phase. With decreasing temperature, the single spin-fluid behavior is recovered below T*˜75 K due to a coherent state among 3 d orbitals. The Kondo liquid scenario is proposed to understand the low-temperature coherent state.

Theory of Dielectric Breakdown in Randomly Inhomogeneous Materials

NASA Astrophysics Data System (ADS)

Gyure, Mark Franklin

1990-01-01

Two models of dielectric breakdown in disordered metal-insulator composites have been developed in an attempt to explain in detail the greatly reduced breakdown electric field observed in these materials. The first model is a two dimensional model in which the composite is treated as a random array of conducting cylinders embedded in an otherwise uniform dielectric background. The two dimensional samples are generated by the Monte Carlo method and a discretized version of the integral form of Laplace's equation is solved to determine the electric field in each sample. Breakdown is modeled as a quasi-static process by which one breakdown at a time occurs at the point of maximum electric field in the system. A cascade of these local breakdowns leads to complete dielectric failure of the system after which the breakdown field can be determined. A second model is developed that is similar to the first in terms of breakdown dynamics, but uses coupled multipole expansions of the electrostatic potential centered at each particle to obtain a more computationally accurate and faster solution to the problem of determining the electric field at an arbitrary point in a random medium. This new algorithm allows extension of the model to three dimensions and treats conducting spherical inclusions as well as cylinders. Successful implementation of this algorithm relies on the use of analytical forms for off-centered expansions of cylindrical and spherical harmonics. Scaling arguments similar to those used in theories of phase transitions are developed for the breakdown field and these arguments are discussed in context with other theories that have been developed to explain the break-down behavior of random resistor and fuse networks. Finally, one of the scaling arguments is used to predict the breakdown field for some samples of solid fuel rocket propellant tested at the China Lake Naval Weapons Center and is found to compare quite well with the experimentally measured breakdown

14 CFR 29.87 - Height-velocity envelope.

Code of Federal Regulations, 2010 CFR

2010-01-01

... Category A engine isolation requirements, the height-velocity envelope for complete power failure must be... 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...

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.

Energizing the last phase of common-envelope removal

NASA Astrophysics Data System (ADS)

Soker, Noam

2017-11-01

We propose a scenario where a companion that is about to exit a common-envelope evolution (CEE) with a giant star accretes mass from the remaining envelope outside its deep orbit and launches jets that facilitate the removal of the remaining envelope. The jets that the accretion disc launches collide with the envelope and form hot bubbles that energize the envelope. Due to gravitational interaction with the envelope, which might reside in a circumbinary disc, the companion migrates farther in, but the inner boundary of the circumbinary disc continues to feed the accretion disc. While near the equatorial plane mass leaves the system at a very low velocity, along the polar directions velocities are very high. When the primary is an asymptotic giant branch star, this type of flow forms a bipolar nebula with very narrow waists. We compare this envelope-removal process with four other last-phase common-envelope-removal processes. We also note that the accreted gas from the envelope outside the orbit in the last phase of the CEE might carry with it angular momentum that is anti-aligned to the orbital angular momentum. We discuss the implications to the possibly anti-aligned spins of the merging black hole event GW170104.

City traffic flow breakdown prediction based on fuzzy rough set

NASA Astrophysics Data System (ADS)

Yang, Xu; Da-wei, Hu; Bing, Su; Duo-jia, Zhang

2017-05-01

In city traffic management, traffic breakdown is a very important issue, which is defined as a speed drop of a certain amount within a dense traffic situation. In order to predict city traffic flow breakdown accurately, in this paper, we propose a novel city traffic flow breakdown prediction algorithm based on fuzzy rough set. Firstly, we illustrate the city traffic flow breakdown problem, in which three definitions are given, that is, 1) Pre-breakdown flow rate, 2) Rate, density, and speed of the traffic flow breakdown, and 3) Duration of the traffic flow breakdown. Moreover, we define a hazard function to represent the probability of the breakdown ending at a given time point. Secondly, as there are many redundant and irrelevant attributes in city flow breakdown prediction, we propose an attribute reduction algorithm using the fuzzy rough set. Thirdly, we discuss how to predict the city traffic flow breakdown based on attribute reduction and SVM classifier. Finally, experiments are conducted by collecting data from I-405 Freeway, which is located at Irvine, California. Experimental results demonstrate that the proposed algorithm is able to achieve lower average error rate of city traffic flow breakdown prediction.

Testing the Definition of the ESC Envelope

NASA Technical Reports Server (NTRS)

Vincent, Mark A.

2017-01-01

The previous effort, including a successful Change Control Request, addressed shrinking the size of the Earth Science Constellations' (ESC) Envelope by reducing the Margin. Fundamental to the purpose of the Envelope is the case where the argument of perigee of the secondary object circulates from 90 degrees to 270 degrees. This ("outside of the envelope, always outside the envelope") case was tested both numerically in a spreadsheet and analytically. Results showed how it is important to include the fact that a secondary with a different semi-major axis has a different frozen eccentricity value.

Bulk charging and breakdown in electron-irradiated polymers

NASA Technical Reports Server (NTRS)

Frederickson, A. R.

1981-01-01

High energy electron irradiations were performed in an experimental and theoretical study of ten common polymers. Breakdowns were monitored by measuring currents between the electrodes on each side of the planar samples. Sample currents as a function of time during irradiation are compared with theory. Breakdowns are correlated with space charge electric field strength and polarity. Major findings include evidence that all polymers tested broke down, breakdowns remove negligible bulk charge and no breakdowns are seen below 20 million V/m.

Study of RF breakdown and multipacting in accelerator components

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pande, Manjiri; Singh, P., E-mail: manjiri@barc.gov.in, E-mail: psingh@barc.gov.in

2014-07-01

Radio frequency (RF) structures that are part of accelerators and energy sources, operate with sinusoidally varying electromagnetic fields under high RF energy. Here, RF breakdown and multipacting take place in RF structures and limit their performance. Electron field emission processes in a RF structure are precursors for breakdown processes. RF breakdown is a major phenomena affecting and causing the irreversible damage to RF structures. Breakdown rate and the damage induced by the breakdowns are its important properties. The damage is related to power absorbed during breakdown, while the breakdown rate is determined by the amplitudes of surface electric and magneticmore » fields, geometry, metal surface preparation and conditioning history. It limits working power and produces irreversible surface damage. The breakdown limit depends on the RF circuit, structure geometry, RF frequency, input RF power, pulse width, materials used, surface processing technique and surface electric and magnetic fields. Multipactor (MP) is a low power, electron multiplication based resonance breakdown phenomenon in vacuum and is often observed in RF structures. A multipactor discharge is undesirable, as it can create a reactive component that detunes the resonant cavities and components, generates noise in communication system and induces gas desorption from the conductor surfaces. In RF structures, certain conditions are required to generate multipacting. (author)« less

Laser Induced Breakdown Spectroscopy (LIBS)

DTIC Science & Technology

2010-03-31

mass spectrometry and laser induced breakdown spectroscopy, Spe T Trejos, A Flores and JR. Almirall, Micro-spectrochemical analysis of document paper...and gel inks by laser ablation inductively coupled plasma mass spectrometry and laser induced breakdown spectroscopy, Spectrochimica Acta Part B...abstracts): 1. *Schenk, E.R. “Elemental analysis of unprocessed cotton by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) and laser

Characteristics of edge breakdowns on Teflon samples

NASA Technical Reports Server (NTRS)

Yadlowsky, E. J.; Hazelton, R. C.; Churchill, R. J.

1980-01-01

The characteristics of electrical discharges induced on silverbacked Teflon samples irradiated by a monoenergetic electron beam have been studied under controlled laboratory conditions. Measurements of breakdown threshold voltages indicate a marked anisotropy in the electrical breakdown properties of Teflon: differences of up to 10 kV in breakdown threshold voltage are observed depending on the sample orientation. The material anisotropy can be utilized in spacecraft construction to reduce the magnitude of discharge currents.

Integrity of the Linker of Nucleoskeleton and Cytoskeleton Is Required for Efficient Herpesvirus Nuclear Egress.

PubMed

Klupp, Barbara G; Hellberg, Teresa; Granzow, Harald; Franzke, Kati; Dominguez Gonzalez, Beatriz; Goodchild, Rose E; Mettenleiter, Thomas C

2017-10-01

Herpesvirus capsids assemble in the nucleus, while final virion maturation proceeds in the cytoplasm. This requires that newly formed nucleocapsids cross the nuclear envelope (NE), which occurs by budding at the inner nuclear membrane (INM), release of the primary enveloped virion into the perinuclear space (PNS), and subsequent rapid fusion with the outer nuclear membrane (ONM). During this process, the NE remains intact, even at late stages of infection. In addition, the spacing between the INM and ONM is maintained, as is that between the primary virion envelope and nuclear membranes. The linker of nucleoskeleton and cytoskeleton (LINC) complex consists of INM proteins with a luminal SUN (Sad1/UNC-84 homology) domain connected to ONM proteins with a KASH (Klarsicht, ANC-1, SYNE homology) domain and is thought to be responsible for spacing the nuclear membranes. To investigate the role of the LINC complex during herpesvirus infection, we generated cell lines constitutively expressing dominant negative (dn) forms of SUN1 and SUN2. Ultrastructural analyses revealed a significant expansion of the PNS and the contiguous intracytoplasmic lumen, most likely representing endoplasmic reticulum (ER), especially in cells expressing dn-SUN2. After infection, primary virions accumulated in these expanded luminal regions, also very distant from the nucleus. The importance of the LINC complex was also confirmed by reduced progeny virus titers in cells expressing dn-SUN2. These data show that the intact LINC complex is required for efficient nuclear egress of herpesviruses, likely acting to promote fusion of primary enveloped virions with the ONM. IMPORTANCE While the viral factors for primary envelopment of nucleocapsids at the inner nuclear membrane are known to the point of high-resolution structures, the roles of cellular components and regulators remain enigmatic. Furthermore, the machinery responsible for fusion with the outer nuclear membrane is unsolved. We show here

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

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

Two breakdown mechanisms in ultrathin alumina barrier magnetic tunnel junctions

NASA Astrophysics Data System (ADS)

Oliver, Bryan; Tuttle, Gary; He, Qing; Tang, Xuefei; Nowak, Janusz

2004-02-01

Two breakdown mechanisms are observed in magnetic tunnel junctions having an ultrathin alumina barrier. The two breakdown mechanisms manifest themselves differently when considering large ensembles of nominally identical devices under different stress conditions. The results suggest that one type of breakdown occurs because of the intrinsic breakdown of a well-formed oxide barrier that can be described by the E model of dielectric breakdown. The other is an extrinsic breakdown related to defects in the barrier rather than the failure of the oxide integrity. The characteristic of extrinsic breakdown suggests that a pre-existing pinhole in the barriers grows in area by means of dissipative (Joule) heating and/or an electric field across the pinhole circumference.

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.

Diffusion in Stellar Envelopes

NASA Astrophysics Data System (ADS)

Seaton, M. J.

Abundances in stellar atmospheres can depend on diffusive movements in much deeper layers of stellar envelopes. Diffusion in envelopes is also of interest in that it can lead to changes in opacities and hence to the structures of stars. For envelopes the radiative accelerations grad can be expressed in terms of quantities which depend only on temperatures, densities and chemical compositions. Computations have been made for the elements C, N, O, Ne, Na, Mg, Al, Si, S, Ar, Ca, Cr, Mn, Fe and Ni and tables are being made generally available through CDS (Strasbourg). Some results from those computations will be presented. The computed values of grad are used to study diffusion of iron-group elements in envelopes of HgMn stars. It is shown that one can define a value tau_0 of the Rosseland-mean optical depth tau such that diffusive movements for tau >= tau_0 do not depend on those for tau <= tau_0. For Cr and Mn we obtain solutions with tau_0 = 1 and are able to make some meaningful comparisons of abundances, as computed and as observed in atmospheres. For Fe we find that diffusive movements are slowed down in regions of T ~= 10^5 K where the dominant ionisation stages are near argon-like. Diffusion of Fe-group elements can produce substantial changes in opacities.

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.

Multi-layered nanoparticles for penetrating the endosome and nuclear membrane via a step-wise membrane fusion process.

PubMed

Akita, Hidetaka; Kudo, Asako; Minoura, Arisa; Yamaguti, Masaya; Khalil, Ikramy A; Moriguchi, Rumiko; Masuda, Tomoya; Danev, Radostin; Nagayama, Kuniaki; Kogure, Kentaro; Harashima, Hideyoshi

2009-05-01

Efficient targeting of DNA to the nucleus is a prerequisite for effective gene therapy. The gene-delivery vehicle must penetrate through the plasma membrane, and the DNA-impermeable double-membraned nuclear envelope, and deposit its DNA cargo in a form ready for transcription. Here we introduce a concept for overcoming intracellular membrane barriers that involves step-wise membrane fusion. To achieve this, a nanotechnology was developed that creates a multi-layered nanoparticle, which we refer to as a Tetra-lamellar Multi-functional Envelope-type Nano Device (T-MEND). The critical structural elements of the T-MEND are a DNA-polycation condensed core coated with two nuclear membrane-fusogenic inner envelopes and two endosome-fusogenic outer envelopes, which are shed in stepwise fashion. A double-lamellar membrane structure is required for nuclear delivery via the stepwise fusion of double layered nuclear membrane structure. Intracellular membrane fusions to endosomes and nuclear membranes were verified by spectral imaging of fluorescence resonance energy transfer (FRET) between donor and acceptor fluorophores that had been dually labeled on the liposome surface. Coating the core with the minimum number of nucleus-fusogenic lipid envelopes (i.e., 2) is essential to facilitate transcription. As a result, the T-MEND achieves dramatic levels of transgene expression in non-dividing cells.

Breakdown phenomena in radio-frequency helium microdischarges

NASA Astrophysics Data System (ADS)

Radmilovic-Radjenovic, M.; Radjenovic, B.; Nina, A.

2008-07-01

In this paper, the Kihara equation has been applied in order to determine the breakdown voltage in helium rf microdischarges. It was found that the Kihara equation, with modified moleculer constants, describes the breakdown process well even for gaps of the order of a few millimeters. A good agreement between numerical solutions of the Kihara equation and the available experimental data reveals that the breakdown voltages depend on the pd product and vary substantially with changes in rf frequencies.

Efficient common-envelope ejection through dust-driven winds

NASA Astrophysics Data System (ADS)

Glanz, Hila; Perets, Hagai B.

2018-04-01

Common-envelope evolution (CEE) is the short-lived phase in the life of an interacting binary-system during which two stars orbit inside a single shared envelope. Such evolution is thought to lead to the inspiral of the binary, the ejection of the extended envelope and the formation of a remnant short-period binary. However, detailed hydrodynamical models of CEE encounter major difficulties. They show that following the inspiral most of the envelope is not ejected; though it expands to larger separations, it remains bound to the binary. Here we propose that dust-driven winds can be produced following the CEE. These can evaporate the envelope following similar processes operating in the ejection of the envelopes of AGB stars. Pulsations in an AGB-star drives the expansion of its envelope, allowing the material to cool down to low temperatures thus enabling dust condensation. Radiation pressure on the dust accelerates it, and through its coupling to the gas it drives winds which eventually completely erode the envelope. We show that the inspiral phase in CE-binaries can effectively replace the role of stellar pulsation and drive the CE expansion to scales comparable with those of AGB stars, and give rise to efficient mass-loss through dust-driven winds.

Lowering relative humidity level increases epidermal protein deimination and drives human filaggrin breakdown.

PubMed

Cau, Laura; Pendaries, Valérie; Lhuillier, Emeline; Thompson, Paul R; Serre, Guy; Takahara, Hidenari; Méchin, Marie-Claire; Simon, Michel

2017-05-01

Deimination (also known as citrullination), the conversion of arginine in a protein to citrulline, is catalyzed by a family of enzymes called peptidylarginine deiminases (PADs). Three PADs are expressed in the epidermis, one of their targets being filaggrin. Filaggrin plays a central role in atopic dermatitis and is a key protein for the epidermal barrier. It aggregates keratins and is cross-linked to cornified envelopes. Following its deimination, it is totally degraded to release free amino acids, contributing to the natural moisturizing factor (NMF). The mechanisms controlling this multistep catabolism in human are unknown. To test whether external humidity plays a role, and investigate the molecular mechanisms involved. Specimens of reconstructed human epidermis (RHEs) produced in humid or dry conditions (>95% or 30-50% relative humidity) were compared. RHEs produced in the dry condition presented structural changes, including a thicker stratum corneum and a larger amount of keratohyalin granules. The transepidermal water loss and the stratum corneum pH were decreased whereas the quantity of NMF was greater. This highly suggested that filaggrin proteolysis was up-regulated. The expression/activity of the proteases involved in filaggrin breakdown did not increase while PAD1 expression and the deimination rate of proteins, including filaggrin, were drastically enhanced. Partial inhibition of PADs with Cl-amidine reversed the effect of dryness on filaggrin breakdown. These results demonstrate the importance of external humidity in the control of human filaggrin metabolism, and suggest that deimination plays a major role in this regulation. Copyright © 2017 Japanese Society for Investigative Dermatology. All rights reserved.

Injection envelope matching in storage rings

DOE Office of Scientific and Technical Information (OSTI.GOV)

Minty, M.G.; Spence, W.L.

1995-05-01

The shape and size of the transverse phase space injected into a storage ring can be deduced from turn-by-turn measurements of the transient behavior of the beam envelope in the ring. Envelope oscillations at 2 x the {beta}-tron frequency indicate the presence of a {beta}-mismatch, while envelope oscillations at the {beta}-tron frequency are the signature of a dispersion function mismatch. Experiments in injection optimization using synchrotron radiation imaging of the beam and a fast-gated camera at the SLC damping rings are reported.

Injection envelope matching in storage rings

NASA Astrophysics Data System (ADS)

Minty, M. G.; Spence, W. L.

1995-05-01

The shape and size of the transverse phase space injected into a storage ring can be deduced from turn-by-turn measurements of the transient behavior of the beam envelope in the ring. Envelope oscillations at 2 x the beta-tron frequency indicate the presence of a beta-mismatch, while envelope oscillations at the beta-tron frequency are the signature of a dispersion function mismatch. Experiments in injection optimization using synchrotron radiation imaging of the beam and a fast-gated camera at the SLC damping rings are reported.

Electrical Breakdown in Water Vapor

NASA Astrophysics Data System (ADS)

Škoro, N.; Marić, D.; Malović, G.; Graham, W. G.; Petrović, Z. Lj.

2011-11-01

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 (˜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.

Envelope Protection for In-Flight Ice Contamination

NASA Technical Reports Server (NTRS)

Gingras, David R.; Barnhart, Billy P.; Ranaudo, Richard J.; Ratvasky, Thomas P.; Morelli, Eugene A.

2010-01-01

Fatal loss-of-control (LOC) accidents have been directly related to in-flight airframe icing. The prototype system presented in this paper directly addresses the need for real-time onboard envelope protection in icing conditions. The combinations of a-priori information and realtime aerodynamic estimations are shown to provide sufficient input for determining safe limits of the flight envelope during in-flight icing encounters. The Icing Contamination Envelope Protection (ICEPro) system has been designed and implemented to identify degradations in airplane performance and flying qualities resulting from ice contamination and provide safe flight-envelope cues to the pilot. Components of ICEPro are described and results from preliminary tests are presented.

Numerical characterization of plasma breakdown in reversed field pinches

NASA Astrophysics Data System (ADS)

Peng, Yanli; Zhang, Ya; Mao, Wenzhe; Yang, Zhoujun; Hu, Xiwei; Jiang, Wei

2018-02-01

In the reversed field pinch, there is considerable interest in investigating the plasma breakdown. Indeed, the plasma formed during the breakdown may have an influence on the confinement and maintenance in the latter process. However, up to now there has been no related work, experimentally or in simulation, regarding plasma breakdown in reversed field pinch (RFP). In order to figure out the physical mechanism behind plasma breakdown, the effects of the toroidal and error magnetic field, as well as the loop voltage have been studied. We find that the error magnetic field cannot be neglected even though it is quite small in the short plasma breakdown phase. As the toroidal magnetic field increases, the averaged electron energy is reduced after plasma breakdown is complete, which is disadvantageous for the latter process. In addition, unlike the voltage limits in the tokamak, loop voltages can be quite high because there are no requirements for superconductivity. Volt-second consumption has a small difference under different loop voltages. The breakdown delay still exists in various loop voltage cases, but it is much shorter compared to that in the tokamak case. In all, successful breakdowns are possible in the RFP under a fairly broad range of parameters.

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.

ELECTRON MICROSCOPY OF MITOSIS IN A RADIOSENSITIVE GIANT AMOEBA

DOE Office of Scientific and Technical Information (OSTI.GOV)

Daniels, E.W.; Roth, L.E.

1962-10-01

Pelomyxa illinoisensis amoebae, the large radiosensitive species, were fixed in OsO/sub 4/ and embedded in either Epon 812 or methacrylate. Ultrastructural morphology is demonstrated in subnuclear structures at interphase and during specific times in mitosis. Evidence of nuclear envelope breakdown and reconstruction is presented. Fragments of nuclear envelope membranes are traced throughout metaphase and anaphase to telophase. Annuli in the nuclear envelope and its fragments are demonstrated. P. illinoisensis is unique in mitochondrial arrangement during metaphase and anaphase-- mitochondria are aligned at the ends of fibrils distal to the chromosomes at the positions occupied by centrioles in other types ofmore » cells; there they remain until the end of anaphase. The radioresistant amoebae, Pelomyxa carolinensis and Amoeba proteus do not have polar mitochondria. P. illinoisensis also differs from the two radioresistant species in nucleolar morphology during interphase, and in the manner of nucleolar dissolution during prophase. On the other hand, helical coils are shown in the interphase nucleoplasm which appear similar to those in the radioresistant amoebae, P. carolinensis and A. groteus. A blister stage in the telophase of P. illinoisensis is described which is interpreted to be the result of a rapid nuclear expansion leading to interphase. This has not been observed in the radioresistant amoebae. (auth)« less

The South Carolina bridge-scour envelope curves

USGS Publications Warehouse

Benedict, Stephen T.; Feaster, Toby D.; Caldwell, Andral W.

2016-09-30

The U.S. Geological Survey, in cooperation with the South Carolina Department of Transportation, conducted a series of three field investigations to evaluate historical, riverine bridge scour in the Piedmont and Coastal Plain regions of South Carolina. These investigations included data collected at 231 riverine bridges, which lead to the development of bridge-scour envelope curves for clear-water and live-bed components of scour. The application and limitations of the South Carolina bridge-scour envelope curves were documented in four reports, each report addressing selected components of bridge scour. The current investigation (2016) synthesizes the findings of these previous reports into a guidance manual providing an integrated procedure for applying the envelope curves. Additionally, the investigation provides limited verification for selected bridge-scour envelope curves by comparing them to field data collected outside of South Carolina from previously published sources. Although the bridge-scour envelope curves have limitations, they are useful supplementary tools for assessing the potential for scour at riverine bridges in South Carolina.

High-Resolution Scanning Electron Microscopy and Immuno-Gold Labeling of the Nuclear Lamina and Nuclear Pore Complex.

PubMed

Goldberg, Martin W

2016-01-01

Scanning electron microscopy (SEM) is a technique used to image surfaces. Field emission SEMs (feSEMs) can resolve structures that are ~0.5-1.5 nm apart. FeSEM, therefore is a useful technique for imaging molecular structures that exist at surfaces such as membranes. The nuclear envelope consists of four membrane surfaces, all of which may be accessible for imaging. Imaging of the cytoplasmic face of the outer membrane gives information about ribosomes and cytoskeletal attachments, as well as details of the cytoplasmic peripheral components of the nuclear pore complex, and is the most easily accessed surface. The nucleoplasmic face of the inner membrane is easily accessible in some cells, such as amphibian oocytes, giving valuable details about the organization of the nuclear lamina and how it interacts with the nuclear pore complexes. The luminal faces of both membranes are difficult to access, but may be exposed by various fracturing techniques. Protocols are presented here for the preparation, labeling, and feSEM imaging of Xenopus laevis oocyte nuclear envelopes.

14 CFR 121.423 - Pilot: Extended Envelope Training.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Pilot: Extended Envelope Training. 121.423... REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Training Program § 121.423 Pilot: Extended Envelope Training. (a) Each certificate holder must include in its approved training program, the extended envelope...

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.;

Vortex breakdown incipience: Theoretical considerations

NASA Technical Reports Server (NTRS)

Berger, Stanley A.; Erlebacher, Gordon

1992-01-01

The sensitivity of the onset and the location of vortex breakdowns in concentrated vortex cores, and the pronounced tendency of the breakdowns to migrate upstream have been characteristic observations of experimental investigations; they have also been features of numerical simulations and led to questions about the validity of these simulations. This behavior seems to be inconsistent with the strong time-like axial evolution of the flow, as expressed explicitly, for example, by the quasi-cylindrical approximate equations for this flow. An order-of-magnitude analysis of the equations of motion near breakdown leads to a modified set of governing equations, analysis of which demonstrates that the interplay between radial inertial, pressure, and viscous forces gives an elliptic character to these concentrated swirling flows. Analytical, asymptotic, and numerical solutions of a simplified non-linear equation are presented; these qualitatively exhibit the features of vortex onset and location noted above.

48 CFR 252.236-7000 - Modification proposals-price breakdown.

Code of Federal Regulations, 2010 CFR

2010-10-01

...-price breakdown. 252.236-7000 Section 252.236-7000 Federal Acquisition Regulations System DEFENSE... CLAUSES Text of Provisions And Clauses 252.236-7000 Modification proposals—price breakdown. As prescribed in 236.570(a), use the following clause: Modification Proposals—Price Breakdown (DEC 1991) (a) The...

Breakdown of Benford's law for birth data

NASA Astrophysics Data System (ADS)

Ausloos, M.; Herteliu, C.; Ileanu, B.

2015-02-01

Long birth time series for Romania are investigated from Benford's law point of view, distinguishing between families with a religious (Orthodox and Non-Orthodox) affiliation. The data extend from Jan. 01, 1905 till Dec. 31, 2001, i.e. over 97 years or 35 429 days. The results point to a drastic breakdown of Benford's law. Some interpretation is proposed, based on the statistical aspects due to population sizes, rather than on human thought constraints when the law breakdown is usually expected. Benford's law breakdown clearly points to natural causes.

Pre-breakdown phenomena and discharges in a gas-liquid system

NASA Astrophysics Data System (ADS)

Tereshonok, D. V.; Babaeva, N. Yu; Naidis, G. V.; Panov, V. A.; Smirnov, B. M.; Son, E. E.

2018-04-01

In this paper, we investigate pre-breakdown and breakdown phenomena in gas-liquid systems. Cavitation void formation and breakdown in bubbles immersed in liquids are studied numerically, while complete breakdown of bubbled water is studied in experiments. It is shown that taking into account the dependence of water dielectric constant on electric field strength plays the same important role for cavitation void appearance under the action of electrostriction forces as the voltage rise time. It is also shown that the initial stage of breakdown in deformed bubbles immersed in liquid strongly depends on spatial orientation of the bubbles relative to the external electric field. The effect of immersed microbubbles, distributed in bulk water, on breakdown time and voltage is studied experimentally. At the breakdown voltage, the slow ‘thermal’ mechanism is changed by the fast ‘streamer-leader’ showing a decrease in breakdown time by two orders of magnitude by introducing microbubbles (0.1% of volumetric gas content) into the water. In addition, the plasma channel is found to pass between nearby microbubbles, exhibiting some ‘guidance’ effect.

Fission yeast Lem2 and Man1 perform fundamental functions of the animal cell nuclear lamina.

PubMed

Gonzalez, Yanira; Saito, Akira; Sazer, Shelley

2012-01-01

In animal cells the nuclear lamina, which consists of lamins and lamin-associated proteins, serves several functions: it provides a structural scaffold for the nuclear envelope and tethers proteins and heterochromatin to the nuclear periphery. In yeast, proteins and large heterochromatic domains including telomeres are also peripherally localized, but there is no evidence that yeast have lamins or a fibrous nuclear envelope scaffold. Nonetheless, we found that the Lem2 and Man1 proteins of the fission yeast Schizosaccharomyces pombe, evolutionarily distant relatives of the Lap2/Emerin/Man1 (LEM) sub-family of animal cell lamin-associated proteins, perform fundamental functions of the animal cell lamina. These integral inner nuclear membrane localized proteins, with nuclear localized DNA binding Helix-Extension-Helix (HEH) domains, impact nuclear envelope structure and integrity, are essential for the enrichment of telomeres at the nuclear periphery and by means of their HEH domains anchor chromatin, most likely transcriptionally repressed heterochromatin, to the nuclear periphery. These data indicate that the core functions of the nuclear lamina are conserved between fungi and animal cells and can be performed in fission yeast, without lamins or other intermediate filament proteins.

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.

Kinetic simulations of gas breakdown in the dense plasma focus

NASA Astrophysics Data System (ADS)

Bennett, N.; Blasco, M.; Breeding, K.; DiPuccio, V.; Gall, B.; Garcia, M.; Gardner, S.; Gatling, J.; Hagen, E. C.; Luttman, A.; Meehan, B. T.; Molnar, S.; O'Brien, R.; Ormond, E.; Robbins, L.; Savage, M.; Sipe, N.; Welch, D. R.

2017-06-01

The first fully kinetic, collisional, and electromagnetic simulations of the breakdown phase of a MA-scale dense plasma focus are described and shown to agree with measured electrical characteristics, including breakdown time. In the model, avalanche ionization is driven by cathode electron emission, and this results in incomplete gas breakdown along the insulator. This reinforces the importance of the conditioning process that creates a metallic layer on the insulator surface. The simulations, nonetheless, help explain the relationship between the gas pressure, the insulator length, and the coaxial gap width. Previously, researchers noted three breakdown patterns related to pressure. Simulation and analytical results show that at low pressures, long ionization path lengths lead to volumetric breakdown, while high pressures lead to breakdown across the relatively small coaxial electrode gap. In an intermediate pressure regime, ionization path lengths are comparable to the insulator length which promotes ideal breakdown along the insulator surface.

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

14 CFR 23.333 - Flight envelope.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Flight envelope. 23.333 Section 23.333... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Structure Flight Loads § 23.333 Flight envelope. (a) General. Compliance with the strength requirements of this subpart must be shown at...

14 CFR 23.333 - Flight envelope.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Flight envelope. 23.333 Section 23.333... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Structure Flight Loads § 23.333 Flight envelope. (a) General. Compliance with the strength requirements of this subpart must be shown at...

14 CFR 23.333 - Flight envelope.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Flight envelope. 23.333 Section 23.333... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Structure Flight Loads § 23.333 Flight envelope. (a) General. Compliance with the strength requirements of this subpart must be shown at...

14 CFR 23.333 - Flight envelope.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Flight envelope. 23.333 Section 23.333... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Structure Flight Loads § 23.333 Flight envelope. (a) General. Compliance with the strength requirements of this subpart must be shown at...

14 CFR 23.333 - Flight envelope.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Flight envelope. 23.333 Section 23.333... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Structure Flight Loads § 23.333 Flight envelope. (a) General. Compliance with the strength requirements of this subpart must be shown at...

Envelope Structures of Gram-Positive Bacteria

PubMed Central

Rajagopal, Mithila; Walker, Suzanne

2016-01-01

Gram-positive organisms, including the pathogens Staphylococcus aureus, Streptococcus pneumoniae and Enterococcus faecalis, have dynamic cell envelopes that mediate interactions with the environment and serve as the first line of defense against toxic molecules. Major components of the cell envelope include peptidoglycan, which is a well-established target for antibiotics, teichoic acids, capsular polysaccharides, surface proteins, and phospholipids. These components can undergo modification to promote pathogenesis, decrease susceptibility to antibiotics and host immune defenses, and enhance survival in hostile environments. This chapter will cover the structure, biosynthesis and important functions of major cell envelope components in Gram-positive bacteria. Possible targets for new antimicrobials will be noted. PMID:26919863

Kinetic simulations of gas breakdown in the dense plasma focus

DOE PAGES

Bennett, N.; Blasco, M.; Breeding, K.; ...

2017-06-09

We describe the first fully-kinetic, collisional, and electromagnetic simulations of the breakdown phase of a MA-scale dense plasma focus and are shown to agree with measured electrical characteristics, including breakdown time. In the model, avalanche ionization is driven by cathode electron emission and this results in incomplete gas breakdown along the insulator. This reinforces the importance of the conditioning process that creates a metallic layer on the insulator surface. The simulations, nonetheless, help explain the relationship between the gas pressure, the insulator length, and the coaxial gap width. In the past, researchers noted three breakdown patterns related to pressure. Simulationmore » and analytic results show that at low pressures, long ionization path lengths lead to volumetric breakdown, while high pressures lead to breakdown across the relatively small coaxial electrode gap. In an intermediate pressure regime, ionization path lengths are comparable to the insulator length which promotes ideal breakdown along the insulator surface.« less

Breakdown Cause and Effect Analysis. Case Study

NASA Astrophysics Data System (ADS)

Biały, Witold; Ružbarský, Juraj

2018-06-01

Every company must ensure that the production process proceeds without interferences. Within this article, the author uses the term "interferences" in reference to unplanned stoppages caused by breakdowns. Unfortunately, usually due to machine operators' mistakes, machines break, which causes stoppages thus generating additional costs for the company. This article shows a cause and effect analysis of a breakdown in a production process. The FMEA as well as quality management tools: the Ishikawa diagram and Pareto chart were used for the analysis. Correction measures were presented which allowed for a significant reduction in the number of stoppages caused by breakdowns.

Thrust Breakdown Characteristics of Conventional Propellers

DTIC Science & Technology

2007-09-01

extends beyond the trailing edge of the blade . These sheets violently collapse as the blade moves out of the wake deficit produced by the hull. This...thrust breakdown, vibration, noise , erosion and blade damage. Propellers operating with enough cavitation to cause thrust breakdown can experience...7 Figure 5. Sensitivity of thrust reduction to harmonic content in wake (Prop 5491) .................. 8 Figure 6. Comparison of

Simulating a binary system that experiences the grazing envelope evolution

NASA Astrophysics Data System (ADS)

Shiber, Sagiv; Soker, Noam

2018-06-01

We conduct three-dimensional hydrodynamical simulations, and show that when a secondary star launches jets while performing spiral-in motion into the envelope of a giant star, the envelope is inflated, some mass is ejected by the jets, and the common envelope phase is postponed. We simulate this grazing envelope evolution (GEE) under the assumption that the secondary star accretes mass from the envelope of the asymptotic giant branch (AGB) star and launches jets. In these simulations we do not yet include the gravitational energy that is released by the spiraling-in binary system. Neither do we include the spinning of the envelope. Considering these omissions, we conclude that our results support the idea that jets might play a crucial role in the common envelope evolution or in preventing it.

Effect of temperature on the electric breakdown strength of dielectric elastomer

NASA Astrophysics Data System (ADS)

Liu, Lei; Chen, Hualing; Sheng, Junjie; Zhang, Junshi; Wang, Yongquan; Jia, Shuhai

2014-03-01

DE (dielectric elastomer) is one of the most promising artificial muscle materials for its large strain over 100% under driving voltage. However, to date, dielectric elastomer actuators (DEAs) are prone to failure due to the temperature-dependent electric breakdown. Previously studies had shown that the electrical breakdown strength was mainly related to the temperature-dependent elasticity modulus and the permittivity of dielectric substances. This paper investigated the influence of ambient temperature on the electric breakdown strength of DE membranes (VHB4910 3M). The electric breakdown experiment of the DE membrane was conducted at different ambient temperatures and pre-stretch levels. The real breakdown strength was obtained by measuring the deformation and the breakdown voltage simultaneously. Then, we found that with the increase of the environment temperature, the electric breakdown strength decreased obviously. Contrarily, the high pre-stretch level led to the large electric breakdown strength. What is more, we found that the deformations of DEs were strongly dependent on the ambient temperature.

Coupled ion redistribution and electronic breakdown in low-alkali boroaluminosilicate glass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Choi, Doo Hyun, E-mail: cooldoo@add.re.kr; Randall, Clive, E-mail: car4@psu.edu; Furman, Eugene, E-mail: euf1@psu.edu

2015-08-28

Dielectrics with high electrostatic energy storage must have exceptionally high dielectric breakdown strength at elevated temperatures. Another important consideration in designing a high performance dielectric is understanding the thickness and temperature dependence of breakdown strengths. Here, we develop a numerical model which assumes a coupled ionic redistribution and electronic breakdown is applied to predict the breakdown strength of low-alkali glass. The ionic charge transport of three likely charge carriers (Na{sup +}, H{sup +}/H{sub 3}O{sup +}, Ba{sup 2+}) was used to calculate the ionic depletion width in low-alkali boroaluminosilicate which can further be used for the breakdown modeling. This model predictsmore » the breakdown strengths in the 10{sup 8}–10{sup 9 }V/m range and also accounts for the experimentally observed two distinct thickness dependent regions for breakdown. Moreover, the model successfully predicts the temperature dependent breakdown strength for low-alkali glass from room temperature up to 150 °C. This model showed that breakdown strengths were governed by minority charge carriers in the form of ionic transport (mostly sodium) in these glasses.« less

The Breakdown Characteristics of the Silicone Oil for Electric Power Apparatus

NASA Astrophysics Data System (ADS)

Yoshida, Hisashi; Yanabu, Satoru

The basic breakdown characteristics of the silicone oil as an insulating medium was studied with aim of realization of electric power apparatus which may be considered to be SF6 free and flame-retarding. As the first step, the impulse breakdown characteristics was measured with three kinds of electrodes whose electric field distributions differed. The breakdown characteristics in silicone oil was explained in relation to stressed oil volume (SOV) and the breakdown stress. At the second step the surface breakdown characteristic for impulse voltage was measured with two kinds of insulators which was set to between plane electrodes. The surface breakdown characteristic for impulse voltage was explained in relation to the ratio of the relative permittivity of oil and insulator. And on the third step, the breakdown characteristics of oil gap after interrupting small capacitive current was studied. In this experiment, the disconnecting switch to interrupt capacitive current was simulated by oil gap after interrupting impulse current, and to measure breakdown characteristics the high impulse voltage was subsequently applied. The breakdown stress in silicone oil after application of impulse current was discussed for insulation recovery characteristics.

Adaptive envelope protection methods for aircraft

NASA Astrophysics Data System (ADS)

Unnikrishnan, Suraj

Carefree handling refers to the ability of a pilot to operate an aircraft without the need to continuously monitor aircraft operating limits. At the heart of all carefree handling or maneuvering systems, also referred to as envelope protection systems, are algorithms and methods for predicting future limit violations. Recently, envelope protection methods that have gained more acceptance, translate limit proximity information to its equivalent in the control channel. Envelope protection algorithms either use very small prediction horizon or are static methods with no capability to adapt to changes in system configurations. Adaptive approaches maximizing prediction horizon such as dynamic trim, are only applicable to steady-state-response critical limit parameters. In this thesis, a new adaptive envelope protection method is developed that is applicable to steady-state and transient response critical limit parameters. The approach is based upon devising the most aggressive optimal control profile to the limit boundary and using it to compute control limits. Pilot-in-the-loop evaluations of the proposed approach are conducted at the Georgia Tech Carefree Maneuver lab for transient longitudinal hub moment limit protection. Carefree maneuvering is the dual of carefree handling in the realm of autonomous Uninhabited Aerial Vehicles (UAVs). Designing a flight control system to fully and effectively utilize the operational flight envelope is very difficult. With the increasing role and demands for extreme maneuverability there is a need for developing envelope protection methods for autonomous UAVs. In this thesis, a full-authority automatic envelope protection method is proposed for limit protection in UAVs. The approach uses adaptive estimate of limit parameter dynamics and finite-time horizon predictions to detect impending limit boundary violations. Limit violations are prevented by treating the limit boundary as an obstacle and by correcting nominal control

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

Breakdown between bare electrodes with an oil-paper interface

NASA Astrophysics Data System (ADS)

Kelley, E. F.; Hebner, R. E., Jr.

1980-06-01

Measurements of the location of electrical breakdown in a composite insulating system were made. For these measurements a paper sample was mounted so that it connected the two electrodes. Electrode structures ranging from plane-plane to sphere-sphere were used. The electrode paper system was tested in oil in an attempt to determine the properties of an oil paper interface. The data indicated that in a carefully prepared system the breakdown will not necessarily occur at the interface. In addition, it was found that the breakdown voltages were not significantly lower for those breakdowns which occurred at the interface than for those which did not. It was noted that if the paper interface was not dried or if many gaseous voids were left in or on the paper, the breakdown will regularly occur at the interface and at a lower voltage.

Numerical simulation of dark envelope soliton in plasma

NASA Astrophysics Data System (ADS)

Wang, Fang-Ping; Han, Juan-fang; Zhang, Jie; Gao, Dong-Ning; Li, Zhong-Zheng; Duan, Wen-Shan; Zhang, Heng

2018-03-01

One-dimensional (1-D) particle-in-cell simulation is used to study the propagation of dark envelop solitons described by the nonlinear Schrödinger equation (NLSE) in electron-ion plasmas. The rational solution of the NLSE is presented, which is proposed as an effective tool for studying the dark envelope soliton in plasma. It is demonstrated by our numerical simulation that there is dark envelope soliton in electron-ion plasmas. The numerical results are in good agreements with the analytical ones from the NLSE which is obtained from the reductive perturbation method. The limitation of the amplitude of dark envelop solitons in plasma is noticed.

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 unless...

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 unless...

Co-dependence between trypanosome nuclear lamina components in nuclear stability and control of gene expression.

PubMed

Maishman, Luke; Obado, Samson O; Alsford, Sam; Bart, Jean-Mathieu; Chen, Wei-Ming; Ratushny, Alexander V; Navarro, Miguel; Horn, David; Aitchison, John D; Chait, Brian T; Rout, Michael P; Field, Mark C

2016-12-15

The nuclear lamina is a filamentous structure subtending the nuclear envelope and required for chromatin organization, transcriptional regulation and maintaining nuclear structure. The trypanosomatid coiled-coil NUP-1 protein is a lamina component functionally analogous to lamins, the major lamina proteins of metazoa. There is little evidence for shared ancestry, suggesting the presence of a distinct lamina system in trypanosomes. To find additional trypanosomatid lamina components we identified NUP-1 interacting proteins by affinity capture and mass-spectrometry. Multiple components of the nuclear pore complex (NPC) and a second coiled-coil protein, which we termed NUP-2, were found. NUP-2 has a punctate distribution at the nuclear periphery throughout the cell cycle and is in close proximity to NUP-1, the NPCs and telomeric chromosomal regions. RNAi-mediated silencing of NUP-2 leads to severe proliferation defects, gross alterations to nuclear structure, chromosomal organization and nuclear envelope architecture. Further, transcription is altered at telomere-proximal variant surface glycoprotein (VSG) expression sites (ESs), suggesting a role in controlling ES expression, although NUP-2 silencing does not increase VSG switching. Transcriptome analysis suggests specific alterations to Pol I-dependent transcription. NUP-1 is mislocalized in NUP-2 knockdown cells and vice versa, implying that NUP-1 and NUP-2 form a co-dependent network and identifying NUP-2 as a second trypanosomatid nuclear lamina component. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

Fundamentals of undervoltage breakdown through the Townsend mechanism

NASA Astrophysics Data System (ADS)

Cooley, James E.

The conditions under which an externally supplied pulse of electrons will induce breakdown in an undervoltaged, low-gain, DC discharge gap are experimentally and theoretically explored. The phenomenon is relevant to fundamental understanding of breakdown physics, to switching applications such as triggered spark gaps and discharge initiation in pulsed-plasma thrusters, and to gas-avalanche particle counters. A dimensionless theoretical description of the phenomenon is formulated and solved numerically. It is found that a significant fraction of the charge on the plates must be injected for breakdown to be achieved at low avalanche-ionization gain, when an electron undergoes fewer than approximately 10 ionizing collisions during one gap transit. It is also found that fewer injected electrons are required as the gain due to electron-impact ionization (alpha process) is increased, or as the sensitivity of the alpha process to electric field is enhanced by decreasing the reduced electric field (electric field divided by pressure, E/p). A predicted insensitivity to ion mobility implies that breakdown is determined during the first electron avalanche when space charge distortion is greatest. A dimensionless, theoretical study of the development of this avalanche reveals a critical value of the reduced electric field to be the value at the Paschen curve minimum divided by 1.6. Below this value, the net result of the electric field distortion is to increase ionization for subsequent avalanches, making undervoltage breakdown possible. Above this value, ionization for subsequent avalanches will be suppressed and undervoltage breakdown is not possible. Using an experimental apparatus in which ultraviolet laser pulses are directed onto a photo-emissive cathode of a parallel-plate discharge gap, it is found that undervoltage breakdown can occur through a Townsend-like mechanism through the buildup of successively larger avalanche generations. The minimum number of injected

Parvoviral nuclear import: bypassing the host nuclear-transport machinery.

PubMed

Cohen, Sarah; Behzad, Ali R; Carroll, Jeffrey B; Panté, Nelly

2006-11-01

The parvovirus Minute virus of mice (MVM) is a small DNA virus that replicates in the nucleus of its host cells. However, very little is known about the mechanisms underlying parvovirus' nuclear import. Recently, it was found that microinjection of MVM into the cytoplasm of Xenopus oocytes causes damage to the nuclear envelope (NE), suggesting that the nuclear-import mechanism of MVM involves disruption of the NE and import through the resulting breaks. Here, fluorescence microscopy and electron microscopy were used to examine the effect of MVM on host-cell nuclear structure during infection of mouse fibroblast cells. It was found that MVM caused dramatic changes in nuclear shape and morphology, alterations of nuclear lamin immunostaining and breaks in the NE of infected cells. Thus, it seems that the unusual nuclear-import mechanism observed in Xenopus oocytes is in fact used by MVM during infection of host cells.

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. © 2010 The Authors. Aging Cell © 2010 Blackwell Publishing Ltd/Anatomical Society of Great Britain and Ireland.

Role of phosphatidylserine receptors in enveloped virus infection.

PubMed

Morizono, Kouki; Chen, Irvin S Y

2014-04-01

We recently demonstrated that a soluble protein, Gas6, can facilitate viral entry by bridging viral envelope phosphatidylserine to Axl, a receptor tyrosine kinase expressed on target cells. The interaction between phosphatidylserine, Gas6, and Axl was originally shown to be a molecular mechanism through which phagocytes recognize phosphatidylserine exposed on dead cells. Since our initial report, several groups have confirmed that Axl/Gas6, as well as other phosphatidylserine receptors, facilitate entry of dengue, West Nile, and Ebola viruses. Virus binding by viral envelope phosphatidylserine is now a viral entry mechanism generalized to many families of viruses. In addition to Axl/Gas6, various molecules are known to recognize phosphatidylserine; however, the effects of these molecules on virus binding and entry have not been comprehensively evaluated and compared. In this study, we examined most of the known human phosphatidylserine-recognizing molecules, including MFG-E8, TIM-1, -3, and -4, CD300a, BAI1, and stabilin-1 and -2, for their abilities to facilitate virus binding and infection. Using pseudotyped lentiviral vectors, we found that a soluble phosphatidylserine-binding protein, MFG-E8, enhances transduction. Cell surface receptors TIM-1 and -4 also enhance virus binding/transduction. The extent of enhancement by these molecules varies, depending on the type of pseudotyping envelope proteins. Mutated MFG-E8, which binds viral envelope phosphatidylserine without bridging virus to cells, but, surprisingly, not annexin V, which has been used to block phagocytosis of dead cells by concealing phosphatidylserine, efficiently blocks these phosphatidylserine-dependent viral entry mechanisms. These results provide insight into understanding the role of viral envelope phosphatidylserine in viral infection. Envelope phosphatidylserine has previously been shown to be important for replication of various envelope viruses, but details of this mechanism(s) were unclear

A Signature in HIV-1 Envelope Leader Peptide Associated with Transition from Acute to Chronic Infection Impacts Envelope Processing and Infectivity

PubMed Central

Asmal, Mohammed; Hellmann, Ina; Liu, Weimin; Keele, Brandon F.; Perelson, Alan S.; Bhattacharya, Tanmoy; Gnanakaran, S.; Daniels, Marcus; Haynes, Barton F.; Korber, Bette T.; Hahn, Beatrice H.; Shaw, George M.; Letvin, Norman L.

2011-01-01

Mucosal transmission of the human immunodeficiency virus (HIV) results in a bottleneck in viral genetic diversity. Gnanakaran and colleagues used a computational strategy to identify signature amino acids at particular positions in Envelope that were associated either with transmitted sequences sampled very early in infection, or sequences sampled during chronic infection. Among the strongest signatures observed was an enrichment for the stable presence of histidine at position 12 at transmission and in early infection, and a recurrent loss of histidine at position 12 in chronic infection. This amino acid lies within the leader peptide of Envelope, a region of the protein that has been shown to influence envelope glycoprotein expression and virion infectivity. We show a strong association between a positively charged amino acid like histidine at position 12 in transmitted/founder viruses with more efficient trafficking of the nascent envelope polypeptide to the endoplasmic reticulum and higher steady-state glycoprotein expression compared to viruses that have a non-basic position 12 residue, a substitution that was enriched among viruses sampled from chronically infected individuals. When expressed in the context of other viral proteins, transmitted envelopes with a basic amino acid position 12 were incorporated at higher density into the virus and exhibited higher infectious titers than did non-signature envelopes. These results support the potential utility of using a computational approach to examine large viral sequence data sets for functional signatures and indicate the importance of Envelope expression levels for efficient HIV transmission. PMID:21876761

A gene delivery system containing nuclear localization signal: Increased nucleus import and transfection efficiency with the assistance of RanGAP1.

PubMed

Chen, Kang; Guo, Lingling; Zhang, Jiulong; Chen, Qing; Wang, Kuanglei; Li, Chenxi; Li, Weinan; Qiao, Mingxi; Zhao, Xiuli; Hu, Haiyang; Chen, Dawei

2017-01-15

In the present report, a degradable gene delivery system (PAMS/DNA/10NLS) containing nucleus location signal peptide (NLS) was prepared. The agarose gel electrophoresis, particle size and zeta potential of PAMS/DNA/10NLS were similar to those of PAMS/DNA, which proved that NLS did not affect the interaction between PAMS and DNA. PAMS/DNA/10NLS exhibited marked extracellular and intracellular degradation under acidic conditions. The degradation was believed to allow NLS to come into contact with importins easily, which was able to mediate the nucleus import. With the help of NLS, PAMS/DNA/10NLS exhibited a higher transfection capability than PAMS/DNA. Moreover, the transfection of PAMS/DNA/10NLS was less dependent on the breakdown of the nucleus envelope than PAMS/DNA. Considering that GTPase-activating protein 1 (RanGAP1) was able to activate the endogenous GTPase, which was necessary for NLS-mediated nucleus import, RanGAP1 overexpressed cells (RanGAP1 cells) were produced. This result showed that RanGAP1 cells had higher GTPase activities than normal cells. Both the nucleus import and transfection efficiency of PAMS/DNA/10NLS were markedly higher in RanGAP1 cells than that in normal cells. The in vivo transfection results also showed that the transfection efficiency of PAMS/DNA/10NLS was higher in RanGAP1 pre-treated mice than that in normal mice. These findings showed that PAMS/DNA/10NLS is a promising gene delivery system with the assistance of RanGAP1. The present report describes the increased transfection efficiency of a degradable gene delivery system (PAMS/DNA/10NLS) containing nuclear location signal (NLS) with the assistance of GTPase-activating protein 1 (RanGAP1). The physicochemical properties of PAMS/DNA/10NLS were similar to those of PAMS/DNA. PAMS/DNA/10NLS exhibited great extracellular and intracellular degradations, which might allow NLS to contact with importins easily. With the help of NLS, PAMS/DNA/10NLS exhibited a higher transfection

Full waveform inversion using envelope-based global correlation norm

NASA Astrophysics Data System (ADS)

Oh, Ju-Won; Alkhalifah, Tariq

2018-05-01

To increase the feasibility of full waveform inversion on real data, we suggest a new objective function, which is defined as the global correlation of the envelopes of modelled and observed data. The envelope-based global correlation norm has the advantage of the envelope inversion that generates artificial low-frequency information, which provides the possibility to recover long-wavelength structure in an early stage. In addition, the envelope-based global correlation norm maintains the advantage of the global correlation norm, which reduces the sensitivity of the misfit to amplitude errors so that the performance of inversion on real data can be enhanced when the exact source wavelet is not available and more complex physics are ignored. Through the synthetic example for 2-D SEG/EAGE overthrust model with inaccurate source wavelet, we compare the performance of four different approaches, which are the least-squares waveform inversion, least-squares envelope inversion, global correlation norm and envelope-based global correlation norm. Finally, we apply the envelope-based global correlation norm on the 3-D Ocean Bottom Cable (OBC) data from the North Sea. The envelope-based global correlation norm captures the strong reflections from the high-velocity caprock and generates artificial low-frequency reflection energy that helps us recover long-wavelength structure of the model domain in the early stages. From this long-wavelength model, the conventional global correlation norm is sequentially applied to invert for higher-resolution features of the model.

Inhibition of Enveloped Viruses Infectivity by Curcumin

PubMed Central

Wen, Hsiao-Wei; Ou, Jun-Lin; Chiou, Shyan-Song; Chen, Jo-Mei; Wong, Min-Liang; Hsu, Wei-Li

2013-01-01

Curcumin, a natural compound and ingredient in curry, has antiinflammatory, antioxidant, and anticarcinogenic properties. Previously, we reported that curcumin abrogated influenza virus infectivity by inhibiting hemagglutination (HA) activity. This study demonstrates a novel mechanism by which curcumin inhibits the infectivity of enveloped viruses. In all analyzed enveloped viruses, including the influenza virus, curcumin inhibited plaque formation. In contrast, the nonenveloped enterovirus 71 remained unaffected by curcumin treatment. We evaluated the effects of curcumin on the membrane structure using fluorescent dye (sulforhodamine B; SRB)-containing liposomes that mimic the viral envelope. Curcumin treatment induced the leakage of SRB from these liposomes and the addition of the influenza virus reduced the leakage, indicating that curcumin disrupts the integrity of the membranes of viral envelopes and of liposomes. When testing liposomes of various diameters, we detected higher levels of SRB leakage from the smaller-sized liposomes than from the larger liposomes. Interestingly, the curcumin concentration required to reduce plaque formation was lower for the influenza virus (approximately 100 nm in diameter) than for the pseudorabies virus (approximately 180 nm) and the vaccinia virus (roughly 335 × 200 × 200 nm). These data provide insights on the molecular antiviral mechanisms of curcumin and its potential use as an antiviral agent for enveloped viruses. PMID:23658730

Effect of chromosome tethering on nuclear organization in yeast.

PubMed

Avşaroğlu, Barış; Bronk, Gabriel; Gordon-Messer, Susannah; Ham, Jungoh; Bressan, Debra A; Haber, James E; Kondev, Jane

2014-01-01

Interphase chromosomes in Saccharomyces cerevisiae are tethered to the nuclear envelope at their telomeres and to the spindle pole body (SPB) at their centromeres. Using a polymer model of yeast chromosomes that includes these interactions, we show theoretically that telomere attachment to the nuclear envelope is a major determinant of gene positioning within the nucleus only for genes within 10 kb of the telomeres. We test this prediction by measuring the distance between the SPB and the silent mating locus (HML) on chromosome III in wild-type and mutant yeast strains that contain altered chromosome-tethering interactions. In wild-type yeast cells we find that disruption of the telomere tether does not dramatically change the position of HML with respect to the SPB, in agreement with theoretical predictions. Alternatively, using a mutant strain with a synthetic tether that localizes an HML-proximal site to the nuclear envelope, we find a significant change in the SPB-HML distance, again as predicted by theory. Our study quantifies the importance of tethering at telomeres on the organization of interphase chromosomes in yeast, which has been shown to play a significant role in determining chromosome function such as gene expression and recombination.

14 CFR 25.333 - Flight maneuvering envelope.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 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 of...

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 of...

Nuclear networking.

PubMed

Xie, Wei; Burke, Brian

2017-07-04

Nuclear lamins are intermediate filament proteins that represent important structural components of metazoan nuclear envelopes (NEs). By combining proteomics and superresolution microscopy, we recently reported that both A- and B-type nuclear lamins form spatially distinct filament networks at the nuclear periphery of mouse fibroblasts. In particular, A-type lamins exhibit differential association with nuclear pore complexes (NPCs). Our studies reveal that the nuclear lamina network in mammalian somatic cells is less ordered and more complex than that of amphibian oocytes, the only other system in which the lamina has been visualized at high resolution. In addition, the NPC component Tpr likely links NPCs to the A-type lamin network, an association that appears to be regulated by C-terminal modification of various A-type lamin isoforms. Many questions remain, however, concerning the structure and assembly of lamin filaments, as well as with their mode of association with other nuclear components such as peripheral chromatin.

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.

Mechanism of vacuum breakdown in radio-frequency accelerating structures

NASA Astrophysics Data System (ADS)

Barengolts, S. A.; Mesyats, V. G.; Oreshkin, V. I.; Oreshkin, E. V.; Khishchenko, K. V.; Uimanov, I. V.; Tsventoukh, M. M.

2018-06-01

It has been investigated whether explosive electron emission may be the initiating mechanism of vacuum breakdown in the accelerating structures of TeV linear electron-positron colliders (Compact Linear Collider). The physical processes involved in a dc vacuum breakdown have been considered, and the relationship between the voltage applied to the diode and the time delay to breakdown has been found. Based on the results obtained, the development of a vacuum breakdown in an rf electric field has been analyzed and the main parameters responsible for the initiation of explosive electron emission have been estimated. The formation of craters on the cathode surface during explosive electron emission has been numerically simulated, and the simulation results are discussed.

Plasma breakdown in a capacitively-coupled radiofrequency argon discharge

NASA Astrophysics Data System (ADS)

Smith, H. B.; Charles, C.; Boswell, R. W.

1998-10-01

Low pressure, capacitively-coupled rf discharges are widely used in research and commercial ventures. Understanding of the non-equilibrium processes which occur in these discharges during breakdown is of interest, both for industrial applications and for a deeper understanding of fundamental plasma behaviour. The voltage required to breakdown the discharge V_brk has long been known to be a strong function of the product of the neutral gas pressure and the electrode seperation (pd). This paper investigates the dependence of V_brk on pd in rf systems using experimental, computational and analytic techniques. Experimental measurements of V_brk are made for pressures in the range 1 -- 500 mTorr and electrode separations of 2 -- 20 cm. A Paschen-style curve for breakdown in rf systems is developed which has the minimum breakdown voltage at a much smaller pd value, and breakdown voltages which are significantly lower overall, than for Paschen curves obtained from dc discharges. The differences between the two systems are explained using a simple analytic model. A Particle-in-Cell simulation is used to investigate a similar pd range and examine the effect of the secondary emission coefficient on the rf breakdown curve, particularly at low pd values. Analytic curves are fitted to both experimental and simulation results.

Low-mass White Dwarfs with Hydrogen Envelopes as a Missing Link in the Tidal Disruption Menu

NASA Astrophysics Data System (ADS)

Law-Smith, Jamie; MacLeod, Morgan; Guillochon, James; Macias, Phillip; Ramirez-Ruiz, Enrico

2017-06-01

We construct a menu of objects that can give rise to bright flares when disrupted by massive black holes (BHs), ranging from planets to evolved stars. Through their tidal disruption, main sequence and evolved stars can effectively probe the existence of otherwise quiescent supermassive BHs, and white dwarfs can probe intermediate mass BHs. Many low-mass white dwarfs possess extended hydrogen envelopes, which allow for the production of prompt flares in disruptive encounters with moderately massive BHs of 105-{10}7 {M}⊙ —masses that may constitute the majority of massive BHs by number. These objects are a missing link in two ways: (1) for probing moderately massive BHs and (2) for understanding the hydrodynamics of the disruption of objects with tenuous envelopes. A flare arising from the tidal disruption of a 0.17 {M}⊙ white dwarf by a {10}5 {M}⊙ {BH} reaches a maximum between 0.6 and 11 days, with a peak fallback rate that is usually super-Eddington and results in a flare that is likely brighter than a typical tidal disruption event. Encounters stripping only the envelope can provide hydrogen-only fallback, while encounters disrupting the core evolve from H- to He-rich fallback. While most tidal disruption candidates observed thus far are consistent with the disruptions of main sequence stars, the rapid timescales of nuclear transients such as Dougie and PTF10iya are naturally explained by the disruption of low-mass white dwarfs. As the number of observed flares continues to increase, the menu presented here will be essential for characterizing nuclear BHs and their environments through tidal disruptions.

48 CFR 14.202-3 - Bid envelopes.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 48 Federal Acquisition Regulations System 1 2010-10-01 2010-10-01 false Bid envelopes. 14.202-3 Section 14.202-3 Federal Acquisition Regulations System FEDERAL ACQUISITION REGULATION CONTRACTING METHODS... envelopes bearing Postage and Fees Paid indicia shall not be distributed with the invitation for bids or...

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

A Real-time Breakdown Prediction Method for Urban Expressway On-ramp Bottlenecks

NASA Astrophysics Data System (ADS)

Ye, Yingjun; Qin, Guoyang; Sun, Jian; Liu, Qiyuan

2018-01-01

Breakdown occurrence on expressway is considered to relate with various factors. Therefore, to investigate the association between breakdowns and these factors, a Bayesian network (BN) model is adopted in this paper. Based on the breakdown events identified at 10 urban expressways on-ramp in Shanghai, China, 23 parameters before breakdowns are extracted, including dynamic environment conditions aggregated with 5-minutes and static geometry features. Different time periods data are used to predict breakdown. Results indicate that the models using 5-10 min data prior to breakdown performs the best prediction, with the prediction accuracies higher than 73%. Moreover, one unified model for all bottlenecks is also built and shows reasonably good prediction performance with the classification accuracy of breakdowns about 75%, at best. Additionally, to simplify the model parameter input, the random forests (RF) model is adopted to identify the key variables. Modeling with the selected 7 parameters, the refined BN model can predict breakdown with adequate accuracy.

The limited role of recombination energy in common envelope removal

NASA Astrophysics Data System (ADS)

Grichener, Aldana; Sabach, Efrat; Soker, Noam

2018-05-01

We calculate the outward energy transport time by convection and photon diffusion in an inflated common envelope and find this time to be shorter than the envelope expansion time. We conclude therefore that most of the hydrogen recombination energy ends in radiation rather than in kinetic energy of the outflowing envelope. We use the stellar evolution code MESA and inject energy inside the envelope of an asymptotic giant branch star to mimic energy deposition by a spiraling-in stellar companion. During 1.7 years the envelope expands by a factor of more than 2. Along the entire evolution the convection can carry the energy very efficiently outwards, to the radius where radiative transfer becomes more efficient. The total energy transport time stays within several months, shorter than the dynamical time of the envelope. Had we included rapid mass loss, as is expected in the common envelope evolution, the energy transport time would have been even shorter. It seems that calculations that assume that most of the recombination energy ends in the outflowing gas might be inaccurate.

Supersonic quasi-axisymmetric vortex breakdown

NASA Technical Reports Server (NTRS)

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

1991-01-01

An extensive computational study of supersonic quasi-axisymmetric vortex breakdown in a configured circular duct is presented. The unsteady, compressible, full Navier-Stokes (NS) equations are used. The NS equations are solved for the quasi-axisymmetric flows using an implicit, upwind, flux difference splitting, finite volume scheme. The quasi-axisymmetric solutions are time accurate and are obtained by forcing the components of the flowfield vector to be equal on two axial planes, which are in close proximity of each other. The effect of Reynolds number, for laminar flows, on the evolution and persistence of vortex breakdown, is studied. Finally, the effect of swirl ration at the duct inlet is investigated.

Laser Radiation-Induced Air Breakdown And Plasma Shielding

NASA Astrophysics Data System (ADS)

Smith, David C.

1981-12-01

Gas breakdown, or the ionization of the air in the path of a high power laser, is a limit on the maximum intensity which can be propagated through the atmosphere. When the threshold for breakdown is exceeded, a high density, high temperature plasma is produced which is opaque to visible and infrared wavelengths and thus absorbs the laser radiation. The threshold in the atmosphere is significantly lower than in pure gases because of laser interaction and vaporization of aerosols. This aspect of air breakdown is discussed in detail. Parametric studies have revealed the scaling laws of breakdown as to wavelength and laser pulse duration, and these will be discussed and compared with existing models. A problem closely related to breakdown is the plasma produc-tion when a high intensity laser interacts with a surface. In this case, the plasma can be beneficial for coupling laser energy into shiny surfaces. The plasma absorbs the laser radiation and reradiates the energy at shorter wavelengths; this shorter wavelength radiation is absorbed by the surface, thus increasing the coupling of energy into the surface. The conditions for the enhancement of laser coupling into surfaces will be discussed, particularly for cw laser beams, an area of recent experimen-tal investigation.

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.

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

Gas breakdown driven by L band short-pulse high-power microwave

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yang Yiming; Yuan Chengwei; Qian Baoliang

2012-12-15

High power microwave (HPM) driven gas breakdown is a major factor in limiting the radiation and transmission of HPM. A method that HPM driven gas breakdown could be obtained by changing the aperture of horn antenna is studied in this paper. Changing the effective aperture of horn antenna can adjust the electric field in near field zone, leading to gas breakdown. With this method, measurements of air and SF{sub 6} breakdowns are carried out on a magnetically insulated transmission-line oscillators, which is capable of generating HPM with pulse duration of 30 ns, and frequency of 1.74 GHz. The typical breakdownmore » waveforms of air and SF{sub 6} are presented. Besides, the breakdown field strengths of the two gases are derived at different pressures. It is found that the effects of air and SF{sub 6} breakdown on the transmission of HPM are different: air breakdown mainly shortens the pulse width of HPM while SF{sub 6} breakdown mainly reduces the peak output power of HPM. The electric field threshold of SF{sub 6} is about 2.4 times larger than that of air. These differences suggest that gas properties have a great effect on the transmission characteristic of HPM in gases.« less

Skin protection and breakdown in the ELBW infant. A national survey.

PubMed

Maguire, D P

1999-08-01

The purpose of this study was to learn how neonatal intensive care unit (NICU) nurses describe the problem of skin breakdown in the extremely low birth weight (ELBW) infant, examine interventions currently used to prevent and treat ELBW infant skin breakdown, and to learn how nurses describe and measure skin breakdown. Questionnaires were sent to 482 NICUs in the United States, and a response was requested from a registered nurse with at least 2 years NICU experience currently employed in the NICU who regularly managed ELBW infants. Questionnaires were returned from 215 NICUs (45%). Analysis revealed that an average of 21% of ELBW infants suffered skin breakdown during the 1st week of life. Nurses who reported the least problems with ELBW infant skin breakdown followed skin-care protocols that limited use of tape and made liberal use of Aquaphor to protect fragile skin. Recommendations from this study include the development of an objective tool to rate skin breakdown and further study of product efficacy used in the treatment of skin breakdown.

Inability of keratinocytes lacking their specific transglutaminase to form cross-linked envelopes: absence of envelopes as a simple diagnostic test for lamellar ichthyosis.

PubMed

Jeon, S; Djian, P; Green, H

1998-01-20

Epidermal keratinocytes, late in their terminal differentiation, form cross-linked envelopes resistant to ionic detergent and reducing agent. Because the cross-linking process is catalyzed by the keratinocyte transglutaminase, the absence of active transglutaminase should result in failure of the keratinocyte to form a cross-linked envelope. Three keratinocyte strains bearing mutations in the keratinocyte transglutaminase were examined: two contained no detectable transglutaminase mRNA and none contained active enzyme. All three were unable to form cross-linked envelopes, either spontaneously in stratified cultures or upon induction with Ca2+. Although stratum corneum of normal humans and scales from patients with different ichthyotic diseases contain cross-linked envelopes, those from patients with transglutaminase-negative lamellar ichthyosis do not. Therefore, the disease due to the absence of transglutaminase may be readily distinguished from other ichthyotic disease by a simple test for cross-linked envelopes.

Redirecting the Cyanobacterial Bicarbonate Transporters BicA and SbtA to the Chloroplast Envelope: Soluble and Membrane Cargos Need Different Chloroplast Targeting Signals in Plants

PubMed Central

Rolland, Vivien; Badger, Murray R.; Price, G. Dean

2016-01-01

Most major crops used for human consumption are C3 plants, which yields are limited by photosynthetic inefficiency. To circumvent this, it has been proposed to implement the cyanobacterial CO2-concentrating mechanism (CCM), principally consisting of bicarbonate transporters and carboxysomes, into plant chloroplasts. As it is currently not possible to recover homoplasmic transplastomic monocots, foreign genes must be introduced in these plants via nuclear transformation. Consequently, it is paramount to ensure that resulting proteins reach the appropriate sub-cellular compartment, which for cyanobacterial transporters BicA and SbtA, is the chloroplast inner-envelope membrane (IEM). At present, targeting signals to redirect large transmembrane proteins from non-chloroplastic organisms to plant chloroplast envelopes are unknown. The goal of this study was to identify such signals, using agrobacteria-mediated transient expression and confocal microscopy to determine the sub-cellular localization of ∼37 GFP-tagged chimeras. Initially, fragments of chloroplast proteins known to target soluble cargos to the stroma were tested for their ability to redirect BicA, but they proved ineffective. Next, different N-terminal regions from Arabidopsis IEM transporters were tested. We demonstrated that the N-terminus of AtHP59, AtPLGG1 or AtNTT1 (92–115 amino acids), containing a cleavable chloroplast transit peptide (cTP) and a membrane protein leader (MPL), was sufficient to redirect BicA or SbtA to the chloroplast envelope. This constitutes the first evidence that nuclear-encoded transmembrane proteins from non-chloroplastic organisms can be targeted to the envelope of plant chloroplasts; a finding which represents an important advance in chloroplast engineering by opening up the door to further manipulation of the chloroplastic envelope. PMID:26973659

Time-dependent dielectric breakdown of plasma-exposed porous organosilicate glass

NASA Astrophysics Data System (ADS)

Nichols, M. T.; Sinha, H.; Wiltbank, C. A.; Antonelli, G. A.; Nishi, Y.; Shohet, J. L.

2012-03-01

Time-dependent dielectric breakdown (TDDB) is a major concern for low-k organosilicate dielectrics. To examine the effect of plasma exposure on TDDB degradation, time-to-breakdown measurements were made on porous SiCOH before and after exposure to plasma. A capillary-array window was used to separate charged particle and vacuum ultraviolet (VUV) photon bombardment. Samples exposed to VUV photons, and a combination of VUV photons and ion bombardment exhibited significant degradation in breakdown time. The samples exposed to VUV photons and ion bombardment showed more degradation in breakdown time in comparison to samples exposed to VUV photons alone.

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.

Nuclear Condensation during Mouse Erythropoiesis Requires Caspase-3-Mediated Nuclear Opening.

PubMed

Zhao, Baobing; Mei, Yang; Schipma, Matthew J; Roth, Eric Wayne; Bleher, Reiner; Rappoport, Joshua Z; Wickrema, Amittha; Yang, Jing; Ji, Peng

2016-03-07

Mammalian erythropoiesis involves chromatin condensation that is initiated in the early stage of terminal differentiation. The mechanisms of chromatin condensation during erythropoiesis are unclear. Here, we show that the mouse erythroblast forms large, transient, and recurrent nuclear openings that coincide with the condensation process. The opening lacks nuclear lamina, nuclear pore complexes, and nuclear membrane, but it is distinct from nuclear envelope changes that occur during apoptosis and mitosis. A fraction of the major histones are released from the nuclear opening and degraded in the cytoplasm. We demonstrate that caspase-3 is required for the nuclear opening formation throughout terminal erythropoiesis. Loss of caspase-3 or ectopic expression of a caspase-3 non-cleavable lamin B mutant blocks nuclear opening formation, histone release, chromatin condensation, and terminal erythroid differentiation. We conclude that caspase-3-mediated nuclear opening formation accompanied by histone release from the opening is a critical step toward chromatin condensation during erythropoiesis in mice. Copyright © 2016 Elsevier Inc. All rights reserved.

Nuclear condensation during mouse erythropoiesis requires caspase-3-mediated nuclear opening

PubMed Central

Zhao, Baobing; Mei, Yang; Schipma, Matthew J; Roth, Eric Wayne; Bleher, Reiner; Rappoport, Joshua Z.; Wickrema, Amittha; Yang, Jing; Ji, Peng

2016-01-01

SUMMARY Mammalian erythropoiesis involves chromatin condensation that is initiated in the early stage of terminal differentiation. The mechanisms of chromatin condensation during erythropoiesis are unclear. Here, we show that the mouse erythroblast forms large, transient, and recurrent nuclear openings that coincide with the condensation process. The opening lacks nuclear lamina, nuclear pore complexes, and nuclear membrane, but it is distinct from nuclear envelope changes that occur during apoptosis and mitosis. A fraction of the major histones are released from the nuclear opening and degraded in the cytoplasm. We demonstrate that caspase-3 is required for the nuclear opening formation throughout terminal erythropoiesis. Loss of caspase-3 or ectopic expression of a caspase-3 non-cleavable lamin B mutant blocks nuclear opening formation, histone release, chromatin condensation, and terminal erythroid differentiation. We conclude that caspase-3-mediated nuclear opening formation accompanied by histone release from the opening is a critical step towards chromatin condensation during erythropoiesis in mice. PMID:26954545

Two LXXLL motifs in the N terminus of Mps1 are required for Mps1 nuclear import during G(2)/M transition and sustained spindle checkpoint responses.

PubMed

Zhang, Xiaojuan; Yin, Qingqing; Ling, Youguo; Zhang, Yanhong; Ma, Runlin; Ma, Qingjun; Cao, Cheng; Zhong, Hui; Liu, Xuedong; Xu, Quanbin

2011-08-15

Spindle assembly checkpoint kinase Mps1 is spatially and temporally regulated during cell cycle progression. Mps1 is predominately localized to the cytosol in interphase cells, whereas it is concentrated on kinetochores in prophase and prometaphase cells. The timing and mechanism of Mps1 redistribution during cell cycle transition is currently poorly understood. Here, we show that Mps1 relocates from the cytosol to the nucleus at the G 2/M boundary prior to nuclear envelope breakdown (NEB). This timely translocation depends on two tandem LXXLL motifs in the N terminus of Mps1, and mutations in either motif abolish Mps1 nuclear accumulation. Furthermore, we found that phosphorylation of Mps1 Ser80 (which is located between the two LXXLL motifs) also plays a role in regulating timely nuclear entry of Mps1. Mps1 that is defective in LXXLL motifs has near wild-type kinase activity. Moreover, the kinase activity of Mps1 appears to be dispensable for nuclear translocation, as inhibition of Mps1 by a highly specific small-molecule inhibitor did not perturb its nuclear entry. Remarkably, translocation-deficient Mps1 can mediate activation of spindle assembly checkpoint response; however, it fails to support a sustained mitotic arrest upon prolonged treatment with nocodazole. The mitotic slippage can be attributed to precocious degradation of Mps1 in the arrested cells. Our studies reveal a novel cell cycle-dependent nuclear translocation signal in the N terminus of Mps1 and suggest that timely nuclear entry could be important for sustaining spindle assembly checkpoint responses.

Two LXXLL motifs in the N terminus of Mps1 are required for Mps1 nuclear import during G2/M transition and sustained spindle checkpoint responses

PubMed Central

Zhang, Xiaojuan; Yin, Qingqing; Ling, Youguo; Zhang, Yanhong; Ma, Runlin; Ma, Qingjun; Cao, Cheng; Zhong, Hui

2011-01-01

Spindle assembly checkpoint kinase Mps1 is spatially and temporally regulated during cell cycle progression. Mps1 is predominately localized to the cytosol in interphase cells, whereas it is concentrated on kinetochores in prophase and prometaphase cells. The timing and mechanism of Mps1 redistribution during cell cycle transition is currently poorly understood. Here, we show that Mps1 relocates from the cytosol to the nucleus at the G2/M boundary prior to nuclear envelope breakdown (NEB). This timely translocation depends on two tandem LXXLL motifs in the N terminus of Mps1, and mutations in either motif abolish Mps1 nuclear accumulation. Furthermore, we found that phosphorylation of Mps1 Ser80 (which is located between the two LXXLL motifs) also plays a role in regulating timely nuclear entry of Mps1. Mps1 that is defective in LXXLL motifs has near wild-type kinase activity. Moreover, the kinase activity of Mps1 appears to be dispensable for nuclear translocation, as inhibition of Mps1 by a highly specific small-molecule inhibitor did not perturb its nuclear entry. Remarkably, translocation-deficient Mps1 can mediate activation of spindle assembly checkpoint response; however, it fails to support a sustained mitotic arrest upon prolonged treatment with nocodazole. The mitotic slippage can be attributed to precocious degradation of Mps1 in the arrested cells. Our studies reveal a novel cell cycle-dependent nuclear translocation signal in the N terminus of Mps1 and suggest that timely nuclear entry could be important for sustaining spindle assembly checkpoint responses. PMID:21778823

Pulsed DF chain-laser breakdown induced by maritime aerosols

NASA Astrophysics Data System (ADS)

Amimoto, S. T.; Whittier, J. S.; Ronkowski, F. G.; Valenzuela, P. R.; Harper, G.

1982-08-01

Thresholds for breakdown induced by liquid and solid aerosols in room air have been measured for a 1 microsec-duration pulsed D2-F2 laser of 3.58 -4.78 micron bandwidth. The DF laser beam was directed into an aerosol chamber that simulated maritime atmospheres on the open sea. Both focus and collimated beams were studied. For a focused beam in which the largest encountered aerosol particles were of 1 to 4 micron diameter, pulsed DF breakdown thresholds were measured to lie in the range 0.6 to 1.8 GW/sq cm. Salt-water aerosol breakdown thresholds for micron-size particles were found to be 15 to 30% higher than the corresponding thresholds for fresh-water particles. For a collimated beam that encountered particle diameters as large as 100 microns, breakdown could not be induced using 0.5- microsec (FWHM) pulses at peak intensities of 59 MW/sq cm. Image converter camera measurements of the radial plasma growth rate of 1.3 cm/microsec (at 1.4 GW/sq cm) were consistent with measurements of the cutoff rate of the transmitted laser beam. Pulsed DF breakdown thresholds of 32 MW/sq cm for 30- micron diameter Al2O3 particles were also measured to permit comparison with the earlier pulsed-HF breakdown results of Lencioni, et al.; the solid-particle threshold measurements agree with the Lencioni data if one assumes that the thresholds for microsecond-duration pulses scales is 1/lambda. An approximate theoretical model of the water particle breakdown process is presented that permits the scaling of the present results to other laser pulse durations, aerosol distributions, and transmission path lengths.

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 the...

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 the...

Nuclear pore complex component MOS7/Nup88 is required for innate immunity and nuclear accumulation of defense regulators in Arabidopsis.

PubMed

Cheng, Yu Ti; Germain, Hugo; Wiermer, Marcel; Bi, Dongling; Xu, Fang; García, Ana V; Wirthmueller, Lennart; Després, Charles; Parker, Jane E; Zhang, Yuelin; Li, Xin

2009-08-01

Plant immune responses depend on dynamic signaling events across the nuclear envelope through nuclear pores. Nuclear accumulation of certain resistance (R) proteins and downstream signal transducers are critical for their functions, but it is not understood how these processes are controlled. Here, we report the identification, cloning, and analysis of Arabidopsis thaliana modifier of snc1,7 (mos7-1), a partial loss-of-function mutation that suppresses immune responses conditioned by the autoactivated R protein snc1 (for suppressor of npr1-1, constitutive 1). mos7-1 single mutant plants exhibit defects in basal and R protein-mediated immunity and in systemic acquired resistance but do not display obvious pleiotropic defects in development, salt tolerance, or plant hormone responses. MOS7 is homologous to human and Drosophila melanogaster nucleoporin Nup88 and resides at the nuclear envelope. In animals, Nup88 attenuates nuclear export of activated NF-kappaB transcription factors, resulting in nuclear accumulation of NF-kappaB. Our analysis shows that nuclear accumulation of snc1 and the defense signaling components Enhanced Disease Susceptibility 1 and Nonexpresser of PR genes 1 is significantly reduced in mos7-1 plants, while nuclear retention of other tested proteins is unaffected. The data suggest that specifically modulating the nuclear concentrations of certain defense proteins regulates defense outputs.

Preserving Envelope Efficiency in Performance Based Code Compliance

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thornton, Brian A.; Sullivan, Greg P.; Rosenberg, Michael I.

2015-06-20

The City of Seattle 2012 Energy Code (Seattle 2014), one of the most progressive in the country, is under revision for its 2015 edition. Additionally, city personnel participate in the development of the next generation of the Washington State Energy Code and the International Energy Code. Seattle has pledged carbon neutrality by 2050 including buildings, transportation and other sectors. The United States Department of Energy (DOE), through Pacific Northwest National Laboratory (PNNL) provided technical assistance to Seattle in order to understand the implications of one potential direction for its code development, limiting trade-offs of long-lived building envelope components less stringentmore » than the prescriptive code envelope requirements by using better-than-code but shorter-lived lighting and heating, ventilation, and air-conditioning (HVAC) components through the total building performance modeled energy compliance path. Weaker building envelopes can permanently limit building energy performance even as lighting and HVAC components are upgraded over time, because retrofitting the envelope is less likely and more expensive. Weaker building envelopes may also increase the required size, cost and complexity of HVAC systems and may adversely affect occupant comfort. This report presents the results of this technical assistance. The use of modeled energy code compliance to trade-off envelope components with shorter-lived building components is not unique to Seattle and the lessons and possible solutions described in this report have implications for other jurisdictions and energy codes.« less

Nuclear Physics in a biological context

NASA Astrophysics Data System (ADS)

Discher, Dennis

2012-02-01

A solid tissue can be soft like fat or brain, stiff like striated muscle and heart, or rigid like bone -- and of course every cell has a nucleus that contributes in some way small or large to tissue mechanics. Indeed, nuclei generally exhibit rheology and plasticity that reflects both the chromatin and the nuclear envelope proteins called lamins, all of which change in differentiation. Profiling of tissue nuclei shows that the nuclear intermediate filament protein Lamin-A/C varies over 30-fold between adult tissues and scales strongly with micro-elasticity of tissue, while other nuclear envelope components such as Lamin-B exhibit small variations. Lamin-A/C has been implicated in aging syndromes that affect muscle and fat but not brain, and we find nuclei in brain-derived cells are indeed dominated by Lamin-B and are much softer than nuclei derived from muscle cells with predominantly Lamin-A/C. In vitro, matrix elasticity can affect expression of nuclear envelope components in adult stem cells, and major changes in Lamin-A/C are indeed shown to direct lineage with lower levels favoring soft tissue and higher levels promoting rigid tissue lineage. Further molecular studies provide evidence that the nucleus transduces physical stress. References: (1) J.D. Pajerowski, K.N. Dahl, F.L. Zhong, P.J. Sammak, and D.E. Discher. Physical plasticity of the nucleus in stem cell differentiation. PNAS 104: 15619-15624 (2007). (2) A. Buxboim, I. Ivanova, and D.E. Discher. Matrix Elasticity, Cytoskeletal Forces, and Physics of the Nucleus: how deeply do cells `feel' outside and in? Journal of Cell Science 123: 297-308 (2010).

Testing Common Envelopes on Double White Dwarf Binaries

NASA Astrophysics Data System (ADS)

Nandez, Jose L. A.; Ivanova, Natalia; Lombardi, James C., Jr.

2015-06-01

The formation of a double white dwarf binary likely involves a common envelope (CE) event between a red giant and a white dwarf (WD) during the most recent episode of Roche lobe overflow mass transfer. We study the role of recombination energy with hydrodynamic simulations of such stellar interactions. We find that the recombination energy helps to expel the common envelope entirely, while if recombination energy is not taken into account, a significant fraction of the common envelope remains bound. We apply our numerical methods to constrain the progenitor system for WD 1101+364 - a double WD binary that has well-measured mass ratio of q=0.87±0.03 and an orbital period of 0.145 days. Our best-fit progenitor for the pre-common envelope donor is a 1.5 ⊙ red giant.

Inability of keratinocytes lacking their specific transglutaminase to form cross-linked envelopes: Absence of envelopes as a simple diagnostic test for lamellar ichthyosis

PubMed Central

Jeon, Saewha; Djian, Philippe; Green, Howard

1998-01-01

Epidermal keratinocytes, late in their terminal differentiation, form cross-linked envelopes resistant to ionic detergent and reducing agent. Because the cross-linking process is catalyzed by the keratinocyte transglutaminase, the absence of active transglutaminase should result in failure of the keratinocyte to form a cross-linked envelope. Three keratinocyte strains bearing mutations in the keratinocyte transglutaminase were examined: two contained no detectable transglutaminase mRNA and none contained active enzyme. All three were unable to form cross-linked envelopes, either spontaneously in stratified cultures or upon induction with Ca2+. Although stratum corneum of normal humans and scales from patients with different ichthyotic diseases contain cross-linked envelopes, those from patients with transglutaminase-negative lamellar ichthyosis do not. Therefore, the disease due to the absence of transglutaminase may be readily distinguished from other ichthyotic diseases by a simple test for cross-linked envelopes. PMID:9435253

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.

A hydrodynamic study of a slow nova outburst. [computerized simulation of thermonuclear runaway in white dwarf envelope

NASA Technical Reports Server (NTRS)

Sparks, W. M.; Starrfield, S.; Truran, J. W.

1978-01-01

The paper reports use of a Lagrangian implicit hydrodynamics computer code incorporating a full nuclear-reaction network to follow a thermonuclear runaway in the hydrogen-rich envelope of a 1.25 solar-mass white dwarf. In this evolutionary sequence the envelope was assumed to be of normal (solar) composition and the resulting outburst closely resembles that of the slow nova HR Del. In contrast, previous CNO-enhanced models resemble fast nova outbursts. The slow-nova model ejects material by radiation pressure when the high luminosity of the rekindled hydrogen shell source exceeds the local Eddington luminosity of the outer layers. This is in contrast to the fast nova outburst where ejection is caused by the decay of the beta(+)-unstable nuclei. Nevertheless, radiation pressure probably plays a major role in ejecting material from the fast nova remnants. Therefore, the sequence from slow to fast novae can be interpreted as a sequence of white dwarfs with increasing amounts of enhanced CNO nuclei in their hydrogen envelopes, although other parameters such as the white-dwarf mass and accretion rate probably contribute to the observed variation between novae.

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.

Conversation Breakdowns in the Audiology Clinic: The Importance of Mutual Gaze

ERIC Educational Resources Information Center

Ekberg, Katie; Hickson, Louise; Grenness, Caitlin

2017-01-01

Background: Conversational breakdowns are a persistent concern for older adults with hearing impairment (HI). Previous studies in experimental settings have investigated potential causes of breakdowns in conversations with a person with HI, and effective strategies for repairing these breakdowns. However, little research has explored the causes of…

Nuclear lamins during gametogenesis, fertilization and early development

NASA Technical Reports Server (NTRS)

Maul, G. G.; Schatten, G.

1986-01-01

The distribution of lamins (described by Gerace, 1978, as major proteins of nuclear envelope) during gametogenesis, fertilization, and early development was investigated in germ cells of a mouse (Mus musculus), an echinoderm (Lytechinus variegatus), and the surf clam (Spisula solidissima) was investigated in order to determine whether the differences detected could be correlated with differences in the function of cells in these stages of the germ cells. In order to monitor the behavior of lamins, the gametes and embryos were labeled with antibodies to lamins A, C, and B extracted from autoimmune sera of patients with scleroderma and Lupus erythematosus. Results indicated that lamin B could be identified in nuclear envelopes on only those nuclei where chromatin is attached and where RNA synthesis takes place.

High-Resolution Laser-Induced Breakdown Spectroscopy used in Homeland Security and Forensic Applications

DOE Office of Scientific and Technical Information (OSTI.GOV)

Martin, Madhavi Z; Wullschleger, Stan D; Vass, Arpad Alexander

The technique of laser-induced breakdown spectroscopy (LIBS) to detect elements for a variety of homeland security applications such as nuclear materials identification and inventory,and forensic applications has been demonstrated. For nuclear materials applications, we detected and profiled metals in coatings that were used to encapsulate nuclear fuel. Multivariate analysis has been successfully employed in the quantification of elements present in treated wood and engineered wood composites. These examples demonstrate that LIBS-based techniques are inherently well suited for diverse environmental applications related to homeland security. Three key advantages are evident: (1) small samples (mg) are sufficient; (2) samples can be analyzedmore » by LIBS very rapidly, and (3) biological materials such as human and animal bones and wood can be analyzed with minimal sample preparation. For forensic applications they have used LIBS to determine differences in animal and human bones. They have also applied this technique in the determination of counterfeit and non-counterfeit currency. They recently applied LIBS in helping to solve a murder case.« less

Application of the Envelope Difference Index to Spectrally Sparse Speech

ERIC Educational Resources Information Center

Souza, Pamela; Hoover, Eric; Gallun, Frederick

2012-01-01

Purpose: Amplitude compression is a common hearing aid processing strategy that can improve speech audibility and loudness comfort but also has the potential to alter important cues carried by the speech envelope. In previous work, a measure of envelope change, the Envelope Difference Index (EDI; Fortune, Woodruff, & Preves, 1994), was moderately…

The breakdown of coordinated decision making in distributed systems.

PubMed

Bearman, Christopher; Paletz, Susannah B F; Orasanu, Judith; Thomas, Matthew J W

2010-04-01

This article aims to explore the nature and resolution of breakdowns in coordinated decision making in distributed safety-critical systems. In safety-critical domains, people with different roles and responsibilities often must work together to make coordinated decisions while geographically distributed. Although there is likely to be a large degree of overlap in the shared mental models of these people on the basis of procedures and experience, subtle differences may exist. Study 1 involves using Aviation Safety Reporting System reports to explore the ways in which coordinated decision making breaks down between pilots and air traffic controllers and the way in which the breakdowns are resolved. Study 2 replicates and extends those findings with the use of transcripts from the Apollo 13 National Aeronautics and Space Administration space mission. Across both studies, breakdowns were caused in part by different types of lower-level breakdowns (or disconnects), which are labeled as operational, informational, or evaluative. Evaluative disconnects were found to be significantly harder to resolve than other types of disconnects. Considering breakdowns according to the type of disconnect involved appears to capture useful information that should assist accident and incident investigators. The current trend in aviation of shifting responsibilities and providing increasingly more information to pilots may have a hidden cost of increasing evaluative disconnects. The proposed taxonomy facilitates the investigation of breakdowns in coordinated decision making and draws attention to the importance of considering subtle differences between participants' mental models when considering complex distributed systems.

Analysis and control of asymmetric vortex flows and supersonic vortex breakdown

NASA Technical Reports Server (NTRS)

Kandil, Osama A.

1991-01-01

Topics relative to the analysis and control of asymmetric vortex flow and supersonic vortex breakdown are discussed. Specific topics include the computation of compressible, quasi-axisymmetric slender vortex flow and breakdown; supersonic quasi-axisymmetric vortex breakdown; and three-dimensional Navier-Stokes asymmetric solutions for cones and cone-cylinder configurations.

Effect of Chromosome Tethering on Nuclear Organization in Yeast

PubMed Central

Avşaroğlu, Barış; Bronk, Gabriel; Gordon-Messer, Susannah; Ham, Jungoh; Bressan, Debra A.; Haber, James E.; Kondev, Jane

2014-01-01

Interphase chromosomes in Saccharomyces cerevisiae are tethered to the nuclear envelope at their telomeres and to the spindle pole body (SPB) at their centromeres. Using a polymer model of yeast chromosomes that includes these interactions, we show theoretically that telomere attachment to the nuclear envelope is a major determinant of gene positioning within the nucleus only for genes within 10 kb of the telomeres. We test this prediction by measuring the distance between the SPB and the silent mating locus (HML) on chromosome III in wild–type and mutant yeast strains that contain altered chromosome-tethering interactions. In wild-type yeast cells we find that disruption of the telomere tether does not dramatically change the position of HML with respect to the SPB, in agreement with theoretical predictions. Alternatively, using a mutant strain with a synthetic tether that localizes an HML-proximal site to the nuclear envelope, we find a significant change in the SPB-HML distance, again as predicted by theory. Our study quantifies the importance of tethering at telomeres on the organization of interphase chromosomes in yeast, which has been shown to play a significant role in determining chromosome function such as gene expression and recombination. PMID:25020108

Electrical breakdown detection system for dielectric elastomer actuators

NASA Astrophysics Data System (ADS)

Ghilardi, Michele; Busfield, James J. C.; Carpi, Federico

2017-04-01

Electrical breakdown of dielectric elastomer actuators (DEAs) is an issue that has to be carefully addressed when designing systems based on this novel technology. Indeed, in some systems electrical breakdown might have serious consequences, not only in terms of interruption of the desired function but also in terms of safety of the overall system (e.g. overheating and even burning). The risk for electrical breakdown often cannot be completely avoided by simply reducing the driving voltages, either because completely safe voltages might not generate sufficient actuation or because internal or external factors might change some properties of the actuator whilst in operation (for example the aging or fatigue of the material, or an externally imposed deformation decreasing the distance between the compliant electrodes). So, there is the clear need for reliable, simple and cost-effective detection systems that are able to acknowledge the occurrence of a breakdown event, making DEA-based devices able to monitor their status and become safer and "selfaware". Here a simple solution for a portable detection system is reported that is based on a voltage-divider configuration that detects the voltage drop at the DEA terminals and assesses the occurrence of breakdown via a microcontroller (Beaglebone Black single-board computer) combined with a real-time, ultra-low-latency processing unit (Bela cape an open-source embedded platform developed at Queen Mary University of London). The system was used to both generate the control signal that drives the actuator and constantly monitor the functionality of the actuator, detecting any breakdown event and discontinuing the supplied voltage accordingly, so as to obtain a safer controlled actuation. This paper presents preliminary tests of the detection system in different scenarios in order to assess its reliability.

Few-cycle carrier envelope phase-dependent stereo detection of electrons.

PubMed

Verhoef, Aart J; Fernández, Alma; Lezius, Matthias; O'Keeffe, Kevin; Uiberacker, Matthias; Krausz, Ferenc

2006-12-01

The spatial distribution of electrons emitted from atoms by few-cycle optical fields is known to be dependent on the carrier envelope phase, i.e., the phase of the field with respect to the pulse envelope. With respect to Paulus et al. [Phys. Rev. Lett.91, 253004 (2003)] we propose a greatly simplified device to measure and control the carrier envelope phase of few-cycle pulses with an accuracy of better than pi/10 based on this principle. We compared different schemes to control the carrier envelope phase of our pulses.

An Introduction to Electrical Breakdown in Dielectrics

DTIC Science & Technology

1985-04-01

PROJECT TASK WORK UNIT ELEMENT NO. NO. NO. NO. 11TI TL E ’tniclude Security Classification) AN INTRODUCTION TO ELECTRICAL 1PERSONAL AUTHOR(S...find themselves working in the area without the benefit of formal coursework. inAlthough the title of the course was High Voltage Engineer- inI titled...this work , "An Introduction to Electrical Breakdown * Phenomena," because breakdown may occur at low voltages when spacecraft systems are considered

Effects of streamwise vortex breakdown on supersonic combustion.

PubMed

Hiejima, Toshihiko

2016-04-01

This paper presents a numerical simulation study of the combustion structure of streamwise vortex breakdown at Mach number 2.48. Hydrogen fuel is injected into a combustor at sonic speed from the rear of a hypermixer strut that can generate streamwise vortices. The results show that the burning behavior is enhanced at the points of the shock waves that are incident on the vortex and therefore the vortex breakdown in the subsonic region occurs due to combustion. The breakdown domain in the mainstream is found to form a flame-holding region suited to combustion and to lead to a stable combustion field with detached flames. In this way, streamwise vortex breakdown has an essential role in combustion enhancement and the formation of flames that hold under supersonic inflow conditions. Finally, the combustion property defined here is shown to coincide with the produced-water mass flow. This property shows that the amount of combustion is saturated at equivalence ratios over 0.4, although there is a slight increase beyond 1.

Efficiency of planetesimal ablation in giant planetary envelopes

NASA Astrophysics Data System (ADS)

Pinhas, Arazi; Madhusudhan, Nikku; Clarke, Cathie

2016-12-01

Observations of exoplanetary spectra are leading to unprecedented constraints on their atmospheric elemental abundances, particularly O/H, C/H, and C/O ratios. Recent studies suggest that elemental ratios could provide important constraints on formation and migration mechanisms of giant exoplanets. A fundamental assumption in such studies is that the chemical composition of the planetary envelope represents the sum-total of compositions of the accreted gas and solids during the formation history of the planet. We investigate the efficiency with which accreted planetesimals ablate in a giant planetary envelope thereby contributing to its composition rather than sinking to the core. From considerations of aerodynamic drag causing `frictional ablation' and the envelope temperature structure causing `thermal ablation', we compute mass ablations for impacting planetesimals of radii 30 m to 1 km for different compositions (ice to iron) and a wide range of velocities and impact angles, assuming spherical symmetry. Icy impactors are fully ablated in the outer envelope for a wide range of parameters. Even for Fe impactors substantial ablation occurs in the envelope for a wide range of sizes and velocities. For example, iron impactors of sizes below ˜0.5 km and velocities above ˜30 km s-1 are found to ablate by ˜60-80 per cent within the outer envelope at pressures below 103 bar due to frictional ablation alone. For deeper pressures (˜107 bar), substantial ablation happens over a wider range of parameters. Therefore, our exploratory study suggests that atmospheric abundances of volatile elements in giant planets reflect their accretion history during formation.

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.

Gravitational Waves from Accreting Neutron Stars Undergoing Common-envelope Inspiral

NASA Astrophysics Data System (ADS)

Holgado, A. Miguel; Ricker, Paul M.; Huerta, E. A.

2018-04-01

The common-envelope phase is a likely formation channel for close binary systems containing compact objects. Neutron stars in common envelopes accrete at a fraction of the Bondi–Hoyle–Lyttleton accretion rate, since the stellar envelope is inhomogeneous, but they may still be able to accrete at hypercritical rates (though not enough to become black holes). We show that common-envelope systems consisting of a neutron star with a massive primary may be gravitational-wave (GW) sources detectable in the Advanced LIGO band as far away as the Magellanic Clouds. To characterize their evolution, we perform orbital integrations using 1D models of 12 M ⊙ and 20 M ⊙ primaries, considering the effects of density gradient on the accretion onto the NS and spin evolution. From the range of possible accretion rates relevant to common-envelope evolution, we find that these systems may be louder GW sources than low-mass X-ray binaries like Sco X-1, which are currently the target of directed searches for continuous GWs. We also find that their strain amplitude signal may allow for novel constraints on the orbital separation and inspiral timescale in common envelopes when combined with pre-common-envelope electromagnetic observations.

Laser-Induced Breakdown Spectroscopy (LIBS) in a Novel Molten Salt Aerosol System.

PubMed

Williams, Ammon N; Phongikaroon, Supathorn

2017-04-01

In the pyrochemical separation of used nuclear fuel (UNF), fission product, rare earth, and actinide chlorides accumulate in the molten salt electrolyte over time. Measuring this salt composition in near real-time is advantageous for operational efficiency, material accountability, and nuclear safeguards. Laser-induced breakdown spectroscopy (LIBS) has been proposed and demonstrated as a potential analytical approach for molten LiCl-KCl salts. However, all the studies conducted to date have used a static surface approach which can lead to issues with splashing, low repeatability, and poor sample homogeneity. In this initial study, a novel molten salt aerosol approach has been developed and explored to measure the composition of the salt via LIBS. The functionality of the system has been demonstrated as well as a basic optimization of the laser energy and nebulizer gas pressure used. Initial results have shown that this molten salt aerosol-LIBS system has a great potential as an analytical technique for measuring the molten salt electrolyte used in this UNF reprocessing technology.

Three-dimensional envelope instability in periodic focusing channels

NASA Astrophysics Data System (ADS)

Qiang, Ji

2018-03-01

The space-charge driven envelope instability can be of great danger in high intensity accelerators and was studied using a two-dimensional (2D) envelope model and three-dimensional (3D) macroparticle simulations before. In this paper, we study the instability for a bunched beam using a three-dimensional envelope model in a periodic solenoid and radio-frequency (rf) focusing channel and a periodic quadrupole and rf focusing channel. This study shows that when the transverse zero current phase advance is below 90 ° , the beam envelope can still become unstable if the longitudinal zero current phase advance is beyond 90 ° . For the transverse zero current phase advance beyond 90 ° , the instability stopband width becomes larger with the increase of the longitudinal focusing strength and even shows different structure from the 2D case when the longitudinal zero current phase advance is beyond 90 ° . Breaking the symmetry of two longitudinal focusing rf cavities and the symmetry between the horizontal focusing and the vertical focusing in the transverse plane in the periodic quadrupole and rf channel makes the instability stopband broader. This suggests that a more symmetric accelerator lattice design might help reduce the range of the envelope instability in parameter space.

Multispectral radiation envelope characteristics of aerial infrared targets

NASA Astrophysics Data System (ADS)

Kou, Tian; Zhou, Zhongliang; Liu, Hongqiang; Yang, Yuanzhi; Lu, Chunguang

2018-07-01

Multispectral detection signals are relatively stable and complementary to single spectral detection signals with deficiencies of severe scintillation and poor anti-interference. To take advantage of multispectral radiation characteristics in the application of infrared target detection, the concept of a multispectral radiation envelope is proposed. To build the multispectral radiation envelope model, the temperature distribution of an aerial infrared target is calculated first. By considering the coupling heat transfer process, the heat balance equation is built by using the node network, and the convective heat transfer laws as a function of target speed are uncovered. Then, the tail flame temperature distribution model is built and the temperature distributions at different horizontal distances are calculated. Second, to obtain the optimal detection angles, envelope models of reflected background multispectral radiation and target multispectral radiation are built. Finally, the envelope characteristics of the aerial target multispectral radiation are analyzed in different wavebands in detail. The results we obtained reflect Wien's displacement law and prove the effectiveness and reasonableness of the envelope model, and also indicate that the major difference between multispectral wavebands is greatly influenced by the target speed. Moreover, optimal detection angles are obtained by numerical simulation, and these are very important for accurate and fast target detection, attack decision-making and developing multispectral detection platforms.

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

An Unconventional Diacylglycerol Kinase That Regulates Phospholipid Synthesis and Nuclear Membrane Growth*♦

PubMed Central

Han, Gil-Soo; O'Hara, Laura; Carman, George M.; Siniossoglou, Symeon

2008-01-01

Changes in nuclear size and shape during the cell cycle or during development require coordinated nuclear membrane remodeling, but the underlying molecular events are largely unknown. We have shown previously that the activity of the conserved phosphatidate phosphatase Pah1p/Smp2p regulates nuclear structure in yeast by controlling phospholipid synthesis and membrane biogenesis at the nuclear envelope. Two screens for novel regulators of phosphatidate led to the identification of DGK1. We show that Dgk1p is a unique diacylglycerol kinase that uses CTP, instead of ATP, to generate phosphatidate. DGK1 counteracts the activity of PAH1 at the nuclear envelope by controlling phosphatidate levels. Overexpression of DGK1 causes the appearance of phosphatidate-enriched membranes around the nucleus and leads to its expansion, without proliferating the cortical endoplasmic reticulum membrane. Mutations that decrease phosphatidate levels decrease nuclear membrane growth in pah1Δ cells. We propose that phosphatidate metabolism is a critical factor determining nuclear structure by regulating nuclear membrane biogenesis. PMID:18458075

Nuclear war effects studied

NASA Astrophysics Data System (ADS)

Widespread starvation resulting from changes in climate in the aftermath of a large-scale nuclear war could kill far more people than would the bombs themselves. That prediction was made in a recent study by the Scientific Committee on Problems of the Environment (SCOPE), an a rm of the International Council of Scientific Unions (ICSU). “Noncombatant and combatant countries alike” would risk mass starvation; SCOPE predicted that all told, 2.5 billion people could die as a result of crop failures and breakdowns in food distribution after a nuclear war.

Influence of ultrasound on the electrical breakdown of transformer oil

NASA Astrophysics Data System (ADS)

Isakaev, E. Kh; Tyuftyaev, A. S.; Gadzhiev, M. Kh; Demirov, N. A.; Akimov, P. L.

2018-01-01

When the transformer oil is exposed to low power ultrasonic waves (< 2 W/cm2) at initial moment the breakdown voltage of transformer oil is reduced relative to the breakdown voltage of pure oil due to degassing and the occurrence of cavitation bubbles. With the increase of sonication time the breakdown voltage also increases, nonlinearly. The experimental data indicate the possibility of using ultrasonic waves of low power for degassing of transformer oil.

Scaling and volatility of breakouts and breakdowns in stock price dynamics.

PubMed

Liu, Lu; Wei, Jianrong; Huang, Jiping

2013-01-01

Because the movement of stock prices is not only ubiquitous in financial markets but also crucial for investors, extensive studies have been done to understand the law behind it. In particular, since the financial crisis in 2008, researchers have a more interest in investigating large market volatilities in order to grasp changing market trends. In this work, we analyze the breakouts and breakdowns of both the Standard & Poor's 500 Index in the US stock market and the Shanghai Composite Index in the Chinese stock market. The breakout usually represents an ongoing upward trend in technical analysis while the breakdown represents an ongoing downward trend. Based on the renormalization method, we introduce two parameters to quantize breakouts and breakdowns, respectively. We discover scaling behavior, characterized by power-law distributions for both the breakouts and breakdowns in the two financial markets with different power-law exponents, which reflect different market volatilities. In detail, the market volatility for breakdowns is usually larger than that for breakouts. Moreover, as an emerging market, the Chinese stock market has larger market volatilities for both the breakouts and breakdowns than the US stock market (a mature market). Further, the short-term volatilities show similar features for both the US stock market and the Chinese stock market. However, the medium-term volatilities in the US stock market are almost symmetrical for the breakouts and breakdowns, whereas those in the Chinese stock market appear to be asymmetrical for the breakouts and breakdowns. The methodology presented here provides a way to understand scaling and hence volatilities of breakouts and breakdowns in stock price dynamics. Our findings not only reveal the features of market volatilities but also make a comparison between mature and emerging financial markets.

Scaling and Volatility of Breakouts and Breakdowns in Stock Price Dynamics

PubMed Central

Liu, Lu; Wei, Jianrong; Huang, Jiping

2013-01-01

Background Because the movement of stock prices is not only ubiquitous in financial markets but also crucial for investors, extensive studies have been done to understand the law behind it. In particular, since the financial crisis in 2008, researchers have a more interest in investigating large market volatilities in order to grasp changing market trends. Methodology/Principal Findings In this work, we analyze the breakouts and breakdowns of both the Standard & Poor’s 500 Index in the US stock market and the Shanghai Composite Index in the Chinese stock market. The breakout usually represents an ongoing upward trend in technical analysis while the breakdown represents an ongoing downward trend. Based on the renormalization method, we introduce two parameters to quantize breakouts and breakdowns, respectively. We discover scaling behavior, characterized by power-law distributions for both the breakouts and breakdowns in the two financial markets with different power-law exponents, which reflect different market volatilities. In detail, the market volatility for breakdowns is usually larger than that for breakouts. Moreover, as an emerging market, the Chinese stock market has larger market volatilities for both the breakouts and breakdowns than the US stock market (a mature market). Further, the short-term volatilities show similar features for both the US stock market and the Chinese stock market. However, the medium-term volatilities in the US stock market are almost symmetrical for the breakouts and breakdowns, whereas those in the Chinese stock market appear to be asymmetrical for the breakouts and breakdowns. Conclusions/Signicance The methodology presented here provides a way to understand scaling and hence volatilities of breakouts and breakdowns in stock price dynamics. Our findings not only reveal the features of market volatilities but also make a comparison between mature and emerging financial markets. PMID:24376577

Anticipating electrical breakdown in dielectric elastomer actuators

NASA Astrophysics Data System (ADS)

Muffoletto, Daniel P.; Burke, Kevin M.; Zirnheld, Jennifer L.

2013-04-01

The output strain of a dielectric elastomer actuator is directly proportional to the square of its applied electric field. However, since the likelihood of electric breakdown is elevated with an increased applied field, the maximum operating electric field of the dielectric elastomer is significantly derated in systems employing these actuators so that failure due to breakdown remains unlikely even as the material ages. In an effort to ascertain the dielectric strength so that stronger electric fields can be applied, partial discharge testing is used to assess the health of the actuator by detecting the charge that is released when localized instances of breakdown partially bridge the insulator. Pre-stretched and unstretched samples of VHB4910 tape were submerged in dielectric oil to remove external sources of partial discharges during testing, and the partial discharge patterns were recorded just before failure of the dielectric sample.

A domain unique to plant RanGAP is responsible for its targeting to the plant nuclear rim

PubMed Central

Rose, Annkatrin; Meier, Iris

2001-01-01

Ran is a small signaling GTPase that is involved in nucleocytoplasmic transport. Two additional functions of animal Ran in the formation of spindle asters and the reassembly of the nuclear envelope in mitotic cells have been recently reported. In contrast to Ras or Rho, Ran is not associated with membranes. Instead, the spatial sequestering of its accessory proteins, the Ran GTPase-activating protein RanGAP and the nucleotide exchange factor RCC1, appears to define the local concentration of RanGTP vs. RanGDP involved in signaling. Mammalian RanGAP is bound to the nuclear pore by a mechanism involving the attachment of small ubiquitin-related modifier protein (SUMO) to its C terminus and the subsequent binding of the SUMOylated domain to the nucleoporin Nup358. Here we show that plant RanGAP utilizes a different mechanism for nuclear envelope association, involving a novel targeting domain that appears to be unique to plants. The N-terminal WPP domain is highly conserved among plant RanGAPs and the small, plant-specific nuclear envelope-associated protein MAF1, but not present in yeast or animal RanGAP. Confocal laser scanning microscopy of green fluorescent protein (GFP) fusion proteins showed that it is necessary for RanGAP targeting and sufficient to target the heterologous protein GFP to the plant nuclear rim. The highly conserved tryptophan and proline residues of the WPP motif are necessary for its function. The 110-aa WPP domain is the first nuclear-envelope targeting domain identified in plants. Its fundamental difference to its mammalian counterpart implies that different mechanisms have evolved in plants and animals to anchor RanGAP at the nuclear surface. PMID:11752475

The Molecular Envelope around the Red Supergiant VY CMa

NASA Astrophysics Data System (ADS)

Muller, S.; Dinh-V-Trung; Lim, J.; Hirano, N.; Muthu, C.; Kwok, S.

2007-02-01

We present millimeter interferometric observations of the molecular envelope around the red supergiant VY CMa with the Submillimeter Array (SMA). The high angular resolution (<2") allows us to derive the structure of the envelope as observed in the 1.3 mm continuum, 12CO(2-1), 13CO(2-1), and SO(65-54) lines emission. The circumstellar envelope is resolved into three components: (1) a dense, compact, and dusty central component, embedded in (2) a more diffuse and extended envelope, and (3) a high-velocity component. We construct a simple model, consisting of a spherically symmetric slowly expanding envelope and bipolar outflows with a wide opening angle (~120°) viewed close to the line of sight (i=15deg). Our model can explain the main features of the SMA data and previous single-dish CO multiline observations. An episode of enhanced mass loss along the bipolar direction is inferred from our modeling. The SMA data provide a better understanding of the complicated morphology seen in the optical/IR high-resolution observations.

Messages from the voices within: regulation of signaling by proteins of the nuclear lamina.

PubMed

Gerace, Larry; Tapia, Olga

2018-01-04

The nuclear lamina (NL) is a protein scaffold lining the nuclear envelope that consists of nuclear lamins and associated transmembrane proteins. It helps to organize the nuclear envelope, chromosomes, and the cytoplasmic cytoskeleton. The NL also has an important role in regulation of signaling, as highlighted by the wide range of human diseases caused by mutations in the genes for NL proteins with associated signaling defects. This review will consider diverse mechanisms for signaling regulation by the NL that have been uncovered recently, including interaction with signaling effectors, modulation of actin assembly and compositional alteration of the NL. Cells with discrete NL mutations often show disruption of multiple signaling pathways, however, and for the most part the mechanistic basis for these complex phenotypes remains to be elucidated. Copyright © 2017. Published by Elsevier Ltd.

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.

Studies on the System Regulating Proton Movement across the Chloroplast Envelope 1

PubMed Central

Peters, Jeanne S.; Berkowitz, Gerald A.

1991-01-01

Studies were undertaken to further characterize the spinach (Spinacea oleracea) chloroplast envelope system, which facilitates H+ movement into and out of the stroma, and, hence, modulates photosynthetic activity by regulating stromal pH. It was demonstrated that high envelope-bound Mg2+ causes stromal acidification and photosynthetic inhibition. High envelope-bound Mg2+ was also found to necessitate the activity of a digitoxinand oligomycin-sensitive ATPase for the maintenance of high stromal pH and photosynthesis in the illuminated chloroplast. In chloroplasts that had high envelope Mg2+ and inhibited envelope ATPase activity, 2-(diethylamino)-N-(2,6-dimethylphenyl)acetamide was found to raise stromal pH and stimulate photosynthesis. 2-(Diethylamino)-N-(2,6-dimethylphenyl)acetamide is an amine anesthetic that is known to act as a monovalent cation channel blocker in mammalian systems. We postulate that the system regulating cation and H+ fluxes across the plastid envelope includes a monovalent cation channel in the envelope, some degree of (envelope-bound Mg2+ modulated) H+ flux linked to monovalent cation antiport, and ATPase-dependent H+ efflux. PMID:16668116

Early-\\x90stage Electrical Breakdown involving Tunneling

NASA Astrophysics Data System (ADS)

Hjalmarson, Harold; Moore, Chris; Schultz, Peter; Bussman, Ezra; Scrymgeour, David; Hopkins, Matt

The early stage of electrical breakdown from a surface is assumed to involve field emission. In real-world applications, the electrical field is often assumed to be increased by geometrical effects. In addition to these enhancement effects, contamination by adsorbates can lead to reductions in the effective work functions. To develop a physics-based understanding beyond the use of these empirical effects, the field emission currents at early times are being computed and measured. The calculations involve a solution of the Boltzmann equation, and the measurements involve a scanning tunneling microscope. Early results from this collaborative theoretical-experimental project will be described in this presentation. The presentation will focus on results for an ideal system with an absence of geometrical effects. Sandia National Laboratories is a multi-mission 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.

Vortex breakdown in closed containers with polygonal cross sections

DOE Office of Scientific and Technical Information (OSTI.GOV)

Naumov, I. V., E-mail: naumov@itp.nsc.ru; Dvoynishnikov, S. V.; Kabardin, I. K.

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 onmore » 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.« less

A numerical study of three-dimensional vortex breakdown

NASA Technical Reports Server (NTRS)

Spall, Robert E.; Ash, Robert L.

1987-01-01

A numerical simulation of bubble-type vortex breakdown using a unique discrete form of the full 3-D, unsteady incompressible Navier-Stokes equations was performed. The Navier-Stokes equations were written in a vorticity-velocity form and the physical problem was not restricted to axisymmetric flow. The problem was parametized on a Rossby- Reynolds-number basis. Utilization of this parameter duo was shown to dictate the form of the free-field boundary condition specification and allowed control of axial breakdown location within the computational domain. The structure of the breakdown bubble was studied through time evolution plots of planar projected velocity vectors as well as through plots of particle traces and vortex lines. These results compared favorably with previous experimental studies. In addition, profiles of all three velocity components are presented at various axial stations and a Fourier analysis was performed to identify the dominant circumferential modes. The dynamics of the breakdown process were studied through plots of axial variation of rate of change of integrated total energy and rate of change of integrated enstrophy, as well as through contour plots of velocity, vorticity and pressure.

Development of Numerical Methods to Estimate the Ohmic Breakdown Scenarios of a Tokamak

NASA Astrophysics Data System (ADS)

Yoo, Min-Gu; Kim, Jayhyun; An, Younghwa; Hwang, Yong-Seok; Shim, Seung Bo; Lee, Hae June; Na, Yong-Su

2011-10-01

The ohmic breakdown is a fundamental method to initiate the plasma in a tokamak. For the robust breakdown, ohmic breakdown scenarios have to be carefully designed by optimizing the magnetic field configurations to minimize the stray magnetic fields. This research focuses on development of numerical methods to estimate the ohmic breakdown scenarios by precise analysis of the magnetic field configurations. This is essential for the robust and optimal breakdown and start-up of fusion devices especially for ITER and its beyond equipped with low toroidal electric field (ET <= 0.3 V/m). A field-line-following analysis code based on the Townsend avalanche theory and a particle simulation code are developed to analyze the breakdown characteristics of actual complex magnetic field configurations including the stray magnetic fields in tokamaks. They are applied to the ohmic breakdown scenarios of tokamaks such as KSTAR and VEST and compared with experiments.

An analysis of spectral envelope-reduction via quadratic assignment problems

NASA Technical Reports Server (NTRS)

George, Alan; Pothen, Alex

1994-01-01

A new spectral algorithm for reordering a sparse symmetric matrix to reduce its envelope size was described. The ordering is computed by associating a Laplacian matrix with the given matrix and then sorting the components of a specified eigenvector of the Laplacian. In this paper, we provide an analysis of the spectral envelope reduction algorithm. We described related 1- and 2-sum problems; the former is related to the envelope size, while the latter is related to an upper bound on the work involved in an envelope Cholesky factorization scheme. We formulate the latter two problems as quadratic assignment problems, and then study the 2-sum problem in more detail. We obtain lower bounds on the 2-sum by considering a projected quadratic assignment problem, and then show that finding a permutation matrix closest to an orthogonal matrix attaining one of the lower bounds justifies the spectral envelope reduction algorithm. The lower bound on the 2-sum is seen to be tight for reasonably 'uniform' finite element meshes. We also obtain asymptotically tight lower bounds for the envelope size for certain classes of meshes.

The impact of heat blanketing envelopes on neutron stars cooling

NASA Astrophysics Data System (ADS)

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

2017-11-01

The goal of this work is to investigate the effects of chemical composition of heat blanketing envelopes of neutron stars on their thermal states and thermal evolution. To this purpose, we employ 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 “onion-like” envelope. For illustration, we apply these results to estimate the internal temperature of the Vela pulsar and to study cooling of neutron stars. We show that uncertainties in the chemical composition of the envelopes can lead up to ~ 2.5 times uncertainty of the internal temperature of the star which significantly complicates theoretical reconstruction of the internal structure of cooling neutron stars from observations of their thermal surface emission.

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 landing...

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 landing...

Phase discrimination of uranium oxides using laser-induced breakdown spectroscopy

NASA Astrophysics Data System (ADS)

Campbell, Keri R.; Wozniak, Nicholas R.; Colgan, James P.; Judge, Elizabeth J.; Barefield, James E.; Kilcrease, David P.; Wilkerson, Marianne P.; Czerwinski, Ken R.; Clegg, Samuel M.

2017-08-01

Nuclear forensics goals for characterizing samples of interest include qualitative and quantitative analysis of major and trace elements, isotopic analysis, phase identification, and physical analysis. These samples may include uranium oxides UO2, U3O8, and UO3, which play an important role in the front end of the nuclear fuel cycle, from mining to fuel fabrication. The focus of this study is to compare the ratios of the intensities of uranium and oxygen emission lines which can be used to distinguish between different uranium oxide materials using Laser-Induced Breakdown Spectroscopy (LIBS). Measurements at varying laser powers were made under an argon atmosphere at 585 Torr to ensure the oxygen emission intensity was originating from the sample, and not from the atmosphere. Fifteen uranium emission lines were used to compare experimental results with theoretical calculations in order to determine the plasma conditions. Using a laser energy of 26 mJ, the uranium lines 591.539 and 682.692 nm provide the highest degree of discrimination between the uranium oxides. The study presented here suggests that LIBS is useful for discriminating uranium oxide phases, UO2, U3O8, and UO3.

NASA Work Breakdown Structure (WBS) Handbook

NASA Technical Reports Server (NTRS)

Terrell, Stefanie M.

2018-01-01

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.

Statistics of vacuum breakdown in the high-gradient and low-rate regime

NASA Astrophysics Data System (ADS)

Wuensch, Walter; Degiovanni, Alberto; Calatroni, Sergio; Korsbäck, Anders; Djurabekova, Flyura; Rajamäki, Robin; Giner-Navarro, Jorge

2017-01-01

In an increasing number of high-gradient linear accelerator applications, accelerating structures must operate with both high surface electric fields and low breakdown rates. Understanding the statistical properties of breakdown occurrence in such a regime is of practical importance for optimizing accelerator conditioning and operation algorithms, as well as of interest for efforts to understand the physical processes which underlie the breakdown phenomenon. Experimental data of breakdown has been collected in two distinct high-gradient experimental set-ups: A prototype linear accelerating structure operated in the Compact Linear Collider Xbox 12 GHz test stands, and a parallel plate electrode system operated with pulsed DC in the kV range. Collected data is presented, analyzed and compared. The two systems show similar, distinctive, two-part distributions of number of pulses between breakdowns, with each part corresponding to a specific, constant event rate. The correlation between distance and number of pulses between breakdown indicates that the two parts of the distribution, and their corresponding event rates, represent independent primary and induced follow-up breakdowns. The similarity of results from pulsed DC to 12 GHz rf indicates a similar vacuum arc triggering mechanism over the range of conditions covered by the experiments.

A study of dielectric breakdown along insulators surrounding conductors in liquid argon

DOE PAGES

Lockwitz, Sarah; Jostlein, Hans

2016-03-22

High voltage breakdown in liquid argon is an important concern in the design of liquid argon time projection chambers, which are often used as neutrino and dark matter detectors. We have made systematic measurements of breakdown voltages in liquid argon along insulators surrounding negative rod electrodes where the breakdown is initiated at the anode. The measurements were performed in an open cryostat filled with commercial grade liquid argon exposed to air, and not the ultra-pure argon required for electron drift. While not addressing all high voltage concerns in liquid argon, these measurements have direct relevance to the design of highmore » voltage feedthroughs especially for averting the common problem of flash-over breakdown. The purpose of these tests is to understand the effects of materials, of breakdown path length, and of surface topology for this geometry and setup. We have found that the only material-specific effects are those due to their permittivity. We have found that the breakdown voltage has no dependence on the length of the exposed insulator. Lastly, a model for the breakdown mechanism is presented that can help inform future designs.« less

PAH formation in carbon-rich circumstellar envelopes

NASA Technical Reports Server (NTRS)

Feigelson, Eric D.; Frenklach, Michael

1989-01-01

While there is growing observational evidence that some fraction of interstellar carbon is in polycyclic aromatic hydrocarbons (PAH's), the mechanisms by which these molecules might be formed have not been extensively studied. A detailed investigation of PAH production in the outflowing molecular envelopes of carbon-rich red giant star is presented. The gasphase kinetics of a chemical reaction mechanism developed to study soot production in hydrocarbon flames is modified to apply in circumstellar environments. It was found that astrophysically significant quantities of PAH's can be formed in carbon star envelopes provided the gas is sufficiently dense and resides for a long time in the temperature range of 900 to 1100 k. The precise yield of PAH's is very sensitive to astronomical parameters of the envelope (e.g., mass loss rate, outflow velocity, and acetylene abundance) and certain poorly determined chemical reaction rates.

Processing of spectral and amplitude envelope of animal vocalizations in the human auditory cortex.

PubMed

Altmann, Christian F; Gomes de Oliveira Júnior, Cícero; Heinemann, Linda; Kaiser, Jochen

2010-08-01

In daily life, we usually identify sounds effortlessly and efficiently. Two properties are particularly salient and of importance for sound identification: the sound's overall spectral envelope and its temporal amplitude envelope. In this study, we aimed at investigating the representation of these two features in the human auditory cortex by using a functional magnetic resonance imaging adaptation paradigm. We presented pairs of sound stimuli derived from animal vocalizations that preserved the time-averaged frequency spectrum of the animal vocalizations and the amplitude envelope. We presented the pairs in four different conditions: (a) pairs with the same amplitude envelope and mean spectral envelope, (b) same amplitude envelope, but different mean spectral envelope, (c) different amplitude envelope, but same mean spectral envelope and (d) both different amplitude envelope and mean spectral envelope. We found fMRI adaptation effects for both the mean spectral envelope and the amplitude envelope of animal vocalizations in overlapping cortical areas in the bilateral superior temporal gyrus posterior to Heschl's gyrus. Areas sensitive to the amplitude envelope extended further anteriorly along the lateral superior temporal gyrus in the left hemisphere, while areas sensitive to the spectral envelope extended further anteriorly along the right lateral superior temporal gyrus. Posterior tonotopic areas within the left superior temporal lobe displayed sensitivity for the mean spectrum. Our findings suggest involvement of primary auditory areas in the representation of spectral cues and encoding of general spectro-temporal features of natural sounds in non-primary posterior and lateral superior temporal cortex. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

Variables associated with family breakdown in healthy and obese/ overweigh adolescents

PubMed Central

de Almeida, Carla Cristina J. N.; Mora, Paula de Oliveira; de Oliveira, Valmir Aparecido; João, Camila Aparecida; João, Carolina Regina; Riccio, Ana Carolina; de Almeida, Carlos Alberto N.

2014-01-01

Objective: To evaluate the presence of family breakdown factors among eutrophic and overweight/obese adolescents. Methods: Cross-sectional study of 242 students aged between 14 and 19 years old, from a public school. Each student was weighed, measured and answered a questionnaire with closed questions addressing the presence of family breakdown factors. The adolescents were divided in two groups: euthophic and overweight/obese. The answers of both groups were compared by Fisher's exact and Mann-Whitney tests. Results: There was no statistically significant difference in the prevalence of the studied factors between the two groups. Comparing the number of positive answers (presence of family breakdown factors) and negative ones (absence of family breakdown factors), no difference was observed between the groups. Conclusions: The inclusion of a control group showed that factors of family breakdown, usually identified as associated with obesity in adolescents, may also be present in eutrophic adolescents. PMID:24676193

N1N8-bis(gamma-glutamyl)spermidine cross-linking in epidermal-cell envelopes. Comparison of cross-link levels in normal and psoriatic cell envelopes.

PubMed Central

Martinet, N; Beninati, S; Nigra, T P; Folk, J E

1990-01-01

N1N8-Bis(gamma-glutamyl)spermidine was found in exhaustive proteolytic digests of isolated cell envelopes from human epidermis at levels comparable with those of epsilon-(gamma-glutamyl)lysine. Significantly higher than normal amounts of these compounds, particularly the bis(gamma-glutamyl)polyamine, were observed in envelopes from afflicted areas (scales) of psoriatic patients. These findings support the notions that N1N8-bis(gamma-glutamyl)spermidine, like epsilon-(gamma-glutamyl)lysine, functions in cell envelopes as an enzyme-generated protein cross-link and stabilizing force and that individuals with the chronic, recurrent skin disease, psoriasis, exhibit in involved epidermis abnormal cell-envelope-protein cross-linking. PMID:2241917

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.

Direction of flagellar rotation in bacterial cell envelopes.

PubMed Central

Ravid, S; Eisenbach, M

1984-01-01

Cell envelopes with functional flagella, isolated from wild-type strains of Escherichia coli and Salmonella typhimurium by formation of spheroplasts with penicillin and subsequent osmotic lysis, demonstrate counterclockwise (CCW)-biased rotation when energized with an electron donor for respiration, DL-lactate. Since the direction of flagellar rotation in bacteria is central to the expression of chemotaxis, we studied the cause of this bias. Our main observations were: (i) spheroplasts acquired a clockwise (CW) bias if instead of being lysed they were further incubated with penicillin; (ii) repellents temporarily caused CW rotation of tethered bacteria and spheroplasts but not of their derived cell envelopes; (iii) deenergizing CW-rotating cheV bacteria by KCN or arsenate treatment caused CCW bias; (iv) cell envelopes isolated from CW-rotating cheC and cheV mutants retained the CW bias, unlike envelopes isolated from cheB and cheZ mutants, which upon cytoplasmic release lost this bias and acquired CCW bias; and (v) an inwardly directed, artificially induced proton current rotated tethered envelopes in CCW direction, but an outwardly directed current was unable to rotate the envelopes. It is concluded that (i) a cytoplasmic constituent is required for the expression of CW rotation (or repression of CCW rotation) in strains which are not defective in the switch; (ii) in the absence of this cytoplasmic constituent, the motor is not reversible in such strains, and it probably is mechanically constricted so as to permit CCW sense of rotation only; (iii) the requirement of CW rotation for ATP is not at the level of the motor or the switch but at one of the preceding functional steps of the chemotaxis machinery; (iv) the cheC and cheV gene products are associated with the cytoplasmic membrane; and (v) direct interaction between the switch-motor system and the repellent sensors is improbable. Images PMID:6370958

The rate of dielectric breakdown weathering of lunar regolith in permanently shadowed regions

NASA Astrophysics Data System (ADS)

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

2017-02-01

Large solar energetic particle events may cause dielectric breakdown in the upper 1 mm of regolith in permanently shadowed regions (PSRs). We estimate how the resulting breakdown weathering compares to meteoroid impact weathering. Although the SEP event rates measured by the Cosmic Ray Telescope for the Effects of Radiation (CRaTER) on the Lunar Reconnaissance Orbiter (LRO) are too low for breakdown to have significantly affected the regolith over the duration of the LRO mission, regolith gardened by meteoroid impacts has been exposed to SEPs for ∼106 yr. Therefore, we estimate that breakdown weathering's production rate of vapor and melt in the coldest PSRs is up to 1.8 - 3.5 ×10-7 kg m-2 yr-1 , which is comparable to that produced by meteoroid impacts. Thus, in PSRs, up to 10-25% of the regolith may have been melted or vaporized by dielectric breakdown. Breakdown weathering could also be consistent with observations of the increased porosity ("fairy castles") of PSR regolith. We also show that it is conceivable that breakdown-weathered material is present in Apollo soil samples. Consequently, breakdown weathering could be an important process within PSRs, and it warrants further investigation.

The nuclear membrane-associated honeycomb structure of the unicellular organism Amoeba proteus: on the search for homologies with the nuclear lamina of metazoa.

PubMed

Schmidt, M; Grossmann, U; Krohne, G

1995-07-01

In the protozoon Amoeba proteus, a complex and highly organized structure with the morphology of a honeycomb is associated with the nucleoplasmic surface of the nuclear membrane. We have tested whether this structure exhibits similarity to the nuclear lamina of metazoic organisms. First, we have shown that the honeycomb layer is composed of 3 to 5 nm thick protein fibrils resistant to treatment with detergent, high salt, and digestion with nucleases, thus possessing properties typical for karyoskeletal elements. However, in contrast to the meshwork of lamin filaments in somatic cells of metazoic organisms, the honeycomb layer is not tightly anchored to the nucleoplasmic side of pore complexes, or to the inner nuclear membrane. Second, in microinjection experiments we investigated whether fluorescently labeled lamins of Xenopus laevis (lamins A and LI) and Drosophila melanogaster (lamin Dmo) were able to associate in vivo with the Amoeba proteus honeycomb structure. In microinjected amoeba these three lamins were efficiently transported into the nucleus, but did not associate with the nuclear envelope. Our results suggest that the Amoeba proteus nuclear envelope, including the honeycomb layer, does not contain proteins exhibiting high homologies to lamins of metazoan species thus preventing the localized assembly of microinjected lamins along the nuclear periphery.

Sensitivity to envelope-based interaural delays at high frequencies: center frequency affects the envelope rate-limitation.

PubMed

Bernstein, Leslie R; Trahiotis, Constantine

2014-02-01

Sensitivity to ongoing interaural temporal disparities (ITDs) was measured using bandpass-filtered pulse trains centered at 4600, 6500, or 9200 Hz. Save for minor differences in the exact center frequencies, those target stimuli were those employed by Majdak and Laback [J. Acoust. Soc. Am. 125, 3903-3913 (2009)]. At each center frequency, threshold ITD was measured for pulse repetition rates ranging from 64 to 609 Hz. The results and quantitative predictions by a cross-correlation-based model indicated that (1) at most pulse repetition rates, threshold ITD increased with center frequency, (2) the cutoff frequency of the putative envelope low-pass filter that determines sensitivity to ITD at high envelope rates appears to be inversely related to center frequency, and (3) both outcomes were accounted for by assuming that, independent of the center frequency, the listeners' decision variable was a constant criterion change in interaural correlation of the stimuli as processed internally. The finding of an inverse relation between center frequency and the envelope rate limitation, while consistent with much prior literature, runs counter to the conclusion reached by Majdak and Laback.

Accoustic Localization of Breakdown in Radio Frequency Accelerating Cavities

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lane, Peter Gwin

Current designs for muon accelerators require high-gradient radio frequency (RF) cavities to be placed in solenoidal magnetic fields. These fields help contain and efficiently reduce the phase space volume of source muons in order to create a usable muon beam for collider and neutrino experiments. In this context and in general, the use of RF cavities in strong magnetic fields has its challenges. It has been found that placing normal conducting RF cavities in strong magnetic fields reduces the threshold at which RF cavity breakdown occurs. To aid the effort to study RF cavity breakdown in magnetic fields, it wouldmore » be helpful to have a diagnostic tool which can localize the source of breakdown sparks inside the cavity. These sparks generate thermal shocks to small regions of the inner cavity wall that can be detected and localized using microphones attached to the outer cavity surface. Details on RF cavity sound sources as well as the hardware, software, and algorithms used to localize the source of sound emitted from breakdown thermal shocks are presented. In addition, results from simulations and experiments on three RF cavities, namely the Aluminum Mock Cavity, the High-Pressure Cavity, and the Modular Cavity, are also given. These results demonstrate the validity and effectiveness of the described technique for acoustic localization of breakdown.« less

Acoustic localization of breakdown in radio frequency accelerating cavities

NASA Astrophysics Data System (ADS)

Lane, Peter

Current designs for muon accelerators require high-gradient radio frequency (RF) cavities to be placed in solenoidal magnetic fields. These fields help contain and efficiently reduce the phase space volume of source muons in order to create a usable muon beam for collider and neutrino experiments. In this context and in general, the use of RF cavities in strong magnetic fields has its challenges. It has been found that placing normal conducting RF cavities in strong magnetic fields reduces the threshold at which RF cavity breakdown occurs. To aid the effort to study RF cavity breakdown in magnetic fields, it would be helpful to have a diagnostic tool which can localize the source of breakdown sparks inside the cavity. These sparks generate thermal shocks to small regions of the inner cavity wall that can be detected and localized using microphones attached to the outer cavity surface. Details on RF cavity sound sources as well as the hardware, software, and algorithms used to localize the source of sound emitted from breakdown thermal shocks are presented. In addition, results from simulations and experiments on three RF cavities, namely the Aluminum Mock Cavity, the High-Pressure Cavity, and the Modular Cavity, are also given. These results demonstrate the validity and effectiveness of the described technique for acoustic localization of breakdown.

Nuclear pore complex tethers to the cytoskeleton.

PubMed

Goldberg, Martin W

2017-08-01

The nuclear envelope is tethered to the cytoskeleton. The best known attachments of all elements of the cytoskeleton are via the so-called LINC complex. However, the nuclear pore complexes, which mediate the transport of soluble and membrane bound molecules, are also linked to the microtubule network, primarily via motor proteins (dynein and kinesins) which are linked, most importantly, to the cytoplasmic filament protein of the nuclear pore complex, Nup358, by the adaptor BicD2. The evidence for such linkages and possible roles in nuclear migration, cell cycle control, nuclear transport and cell architecture are discussed. Copyright © 2017. Published by Elsevier Ltd.

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

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

DC breakdown characteristics of silicone polymer composites for HVDC insulator applications

NASA Astrophysics Data System (ADS)

Han, Byung-Jo; Seo, In-Jin; Seong, Jae-Kyu; Hwang, Young-Ho; Yang, Hai-Won

2015-11-01

Critical components for HVDC transmission systems are polymer insulators, which have stricter requirements that are more difficult to achieve compared to those of HVAC insulators. In this study, we investigated the optimal design of HVDC polymer insulators by using a DC electric field analysis and experiments. The physical properties of the polymer specimens were analyzed to develop an optimal HVDC polymer material, and four polymer specimens were prepared for DC breakdown experiments. Single and reverse polarity breakdown tests were conducted to analyze the effect of temperature on the breakdown strength of the polymer. In addition, electric fields were analyzed via simulations, in which a small-scale polymer insulator model was applied to prevent dielectric breakdown due to electric field concentration, with four DC operating conditions taken into consideration. The experimental results show that the electrical breakdown strength and the electric field distribution exhibit significant differences in relation to different DC polarity transition procedures.

Envelope: interactive software for modeling and fitting complex isotope distributions.

PubMed

Sykes, Michael T; Williamson, James R

2008-10-20

An important aspect of proteomic mass spectrometry involves quantifying and interpreting the isotope distributions arising from mixtures of macromolecules with different isotope labeling patterns. These patterns can be quite complex, in particular with in vivo metabolic labeling experiments producing fractional atomic labeling or fractional residue labeling of peptides or other macromolecules. In general, it can be difficult to distinguish the contributions of species with different labeling patterns to an experimental spectrum and difficult to calculate a theoretical isotope distribution to fit such data. There is a need for interactive and user-friendly software that can calculate and fit the entire isotope distribution of a complex mixture while comparing these calculations with experimental data and extracting the contributions from the differently labeled species. Envelope has been developed to be user-friendly while still being as flexible and powerful as possible. Envelope can simultaneously calculate the isotope distributions for any number of different labeling patterns for a given peptide or oligonucleotide, while automatically summing these into a single overall isotope distribution. Envelope can handle fractional or complete atom or residue-based labeling, and the contribution from each different user-defined labeling pattern is clearly illustrated in the interactive display and is individually adjustable. At present, Envelope supports labeling with 2H, 13C, and 15N, and supports adjustments for baseline correction, an instrument accuracy offset in the m/z domain, and peak width. Furthermore, Envelope can display experimental data superimposed on calculated isotope distributions, and calculate a least-squares goodness of fit between the two. All of this information is displayed on the screen in a single graphical user interface. Envelope supports high-quality output of experimental and calculated distributions in PNG or PDF format. Beyond simply

Nuclear Exodus: Herpesviruses Lead the Way

PubMed Central

Bigalke, Janna M.; Heldwein, Ekaterina E.

2016-01-01

Most DNA viruses replicate in the nucleus and exit it either by passing through the nuclear pores or by rupturing the nuclear envelope. Unusually, herpesviruses have evolved a complex mechanism of nuclear escape whereby nascent capsids bud at the inner nuclear membrane to form perinuclear virions that subsequently fuse with the outer nuclear membrane, releasing capsids into the cytosol. Although this general scheme is accepted in the field, the players and their roles are still debated. Recent studies illuminated critical mechanistic features of this enigmatic process and uncovered surprising parallels with a novel cellular nuclear export process. This review summarizes our current understanding of nuclear egress in herpesviruses, examines the experimental evidence and models, and outlines outstanding questions with the goal of stimulating new research in this area. PMID:27482898

Desaturation of oleoyl groups in envelope membranes from spinach chloroplasts.

PubMed Central

Schmidt, H; Heinz, E

1990-01-01

Envelope membranes isolated from chloroplasts of spinach (Spinacia oleracea) desaturate oleoyl groups in monogalactosyl diacylglycerol to linoleoyl groups. The desaturation requires NADPH in combination with ferredoxin and is not restricted to monogalactosyl diacylglycerol, since it is also observed in biosynthetic intermediates as, for example, in phosphatidic acid. This indicates a certain degree of unspecificity of the oleate desaturase in isolated envelope membranes. Lipid desaturation is another important function of chloroplast envelopes. PMID:11607123

Nuclear transporters in a multinucleated organism: functional and localization analyses in Aspergillus nidulans

PubMed Central

Markina-Iñarrairaegui, Ane; Etxebeste, Oier; Herrero-García, Erika; Araújo-Bazán, Lidia; Fernández-Martínez, Javier; Flores, Jairo A.; Osmani, Stephen A.; Espeso, Eduardo A.

2011-01-01

Nuclear transporters mediate bidirectional macromolecule traffic through the nuclear pore complex (NPC), thus participating in vital processes of eukaryotic cells. A systematic functional analysis in Aspergillus nidulans permitted the identification of 4 essential nuclear transport pathways of a hypothetical number of 14. The absence of phenotypes for most deletants indicates redundant roles for these nuclear receptors. Subcellular distribution studies of these carriers show three main distributions: nuclear, nucleocytoplasmic, and in association with the nuclear envelope. These locations are not specific to predicted roles as exportins or importins but indicate that bidirectional transport may occur coordinately in all nuclei of a syncytium. Coinciding with mitotic NPC rearrangements, transporters dynamically modified their localizations, suggesting supplementary roles to nucleocytoplasmic transport specifically during mitosis. Loss of transportin-SR and Mex/TAP from the nuclear envelope indicates absence of RNA transport during the partially open mitosis of Aspergillus, whereas nucleolar accumulation of Kap121 and Kap123 homologues suggests a role in nucleolar disassembly. This work provides new insight into the roles of nuclear transporters and opens an avenue for future studies of the molecular mechanisms of transport among nuclei within a common cytoplasm, using A. nidulans as a model organism. PMID:21880896

FHL1B Interacts with Lamin A/C and Emerin at the Nuclear Lamina and is Misregulated in Emery-Dreifuss Muscular Dystrophy.

PubMed

Ziat, Esma; Mamchaoui, Kamel; Beuvin, Maud; Nelson, Isabelle; Azibani, Feriel; Spuler, Simone; Bonne, Gisèle; Bertrand, Anne T

2016-11-29

Emery-Dreifuss muscular dystrophy (EDMD) is associated with mutations in EMD and LMNA genes, encoding for the nuclear envelope proteins emerin and lamin A/C, indicating that EDMD is a nuclear envelope disease. We recently reported mutations in FHL1 gene in X-linked EDMD. FHL1 encodes FHL1A, and the two minor isoforms FHL1B and FHL1C. So far, none have been described at the nuclear envelope. To gain insight into the pathophysiology of EDMD, we focused our attention on the poorly characterized FHL1B isoform. The amount and the localisation of FHL1B were evaluated in control and diseased human primary myoblasts using immunofluorescence and western blotting. We found that in addition to a cytoplasmic localization, this isoform strongly accumulated at the nuclear envelope of primary human myoblasts, like but independently of lamin A/C and emerin. During myoblast differentiation, we observed a major reduction of FHL1B protein expression, especially in the nucleus. Interestingly, we found elevated FHL1B expression level in myoblasts from an FHL1-related EDMD patient where the FHL1 mutation only affects FHL1A, as well as in myoblasts from an LMNA-related EDMD patient. Altogether, the specific localization of FHL1B and its modulation in disease-patient's myoblasts confirmed FHL1-related EDMD as a nuclear envelope disease.

Evolution of Space Shuttle Range Safety (RS) Ascent Flight Envelope Design

NASA Technical Reports Server (NTRS)

Brewer, Joan D.

2011-01-01

Ascent flight envelopes are trajectories that define the normal operating region of a space vehicle s position from liftoff until the end of powered flight. They fulfill part of the RS data requirements imposed by the Air Force s 45th Space Wing (45SW) on space vehicles launching from the Eastern Range (ER) in Florida. The 45SW is chartered to protect the public by minimizing risks associated with the inherent hazards of launching a vehicle into space. NASA s Space Shuttle program has launched 130+ manned missions over a 30 year period from the ER. Ascent envelopes were delivered for each of those missions. The 45SW envelope requirements have remained largely unchanged during this time. However, the methodology and design processes used to generate the envelopes have evolved over the years to support mission changes, maintain high data quality, and reduce costs. The evolution of the Shuttle envelope design has yielded lessons learned that can be applied to future endevours. There have been numerous Shuttle ascent design enhancements over the years that have caused the envelope methodology to evolve. One of these Shuttle improvements was the introduction of onboard flight software changes implemented to improve launch probability. This change impacted the preflight nominal ascent trajectory, which is a key element in the RS envelope design. While the early Shuttle nominal trajectories were designed preflight using a representative monthly mean wind, the new software changes involved designing a nominal ascent trajectory on launch day using real-time winds. Because the actual nominal trajectory position was not known until launch day, the envelope analysis had to be customized to account for this nominal trajectory variation in addition to the other envelope components.

Response of gypsy moth larvae to homologous and heterologous nuclear polyhedrosis virus

Treesearch

Kathleen S. Shields; Edward M. Dougherty

1991-01-01

The gypsy moth, Lymantria dispar, is not particularly susceptible to baculoviruses other than the nuclear polyhedrosis virus originally isolated from the species (LdMNPV). The multiple enveloped nuclear polyhedrosis virus of Autographa californica (AcMNPV), a very virulent baculovirus that replicates in a large number of...

The Rate of Dielectric Breakdown Weathering of Lunar Regolith in Permanently Shadowed Regions

NASA Technical Reports Server (NTRS)

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

2016-01-01

Large solar energetic particle events may cause dielectric breakdown in the upper 1 mm of regolith in permanently shadowed regions (PSRs). We estimate how the resulting breakdown weathering compares to meteoroid impact weathering. Although the SEP event rates measured by the Cosmic Ray Telescope for the Effects of Radiation (CRaTER) on the Lunar Reconnaissance Orbiter (LRO) are too low for breakdown to have significantly affected the regolith over the duration of the LRO mission, regolith gardened by meteoroid impacts has been exposed to SEPs for approx.10(exp 6 yr. Therefore, we estimate that breakdown weathering's production rate of vapor and melt in the coldest PSRs is up to 1.8-3. 5 ×10(exp -7) kg/sq m/yr, which is comparable to that produced by meteoroid impacts. Thus, in PSRs, up to 10-25% of the regolith may have been melted or vaporized by dielectric breakdown. Breakdown weathering could also be consistent with observations of the increased porosity ("fairy castles") of PSR regolith. We also show that it is con- ceivable that breakdown-weathered material is present in Apollo soil samples. Consequently, breakdown weathering could be an important process within PSRs, and it warrants further investigation.

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.

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.

Dielectric Barrier Discharges: Pulsed Breakdown, Electrical Characterization and Chemistry

DTIC Science & Technology

2013-06-01

DIELECTRIC BARRIER DISCHARGES : PULSED BREAKDOWN, ELECTRICAL CHARACTERIZATION AND CHEMISTRY  R. Brandenburg, H. Höft, T. Hoder, A. Pipa, R...for pulsed driven Dielectric Barrier Discharges (DBDs) in particular. Fast electrical, optical and spectroscopic methods enable the study of...2. REPORT TYPE N/A 3. DATES COVERED - 4. TITLE AND SUBTITLE Dielectric Barrier Discharges : Pulsed Breakdown, Electrical Characterization

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…

Alternating current breakdown voltage of ice electret

NASA Astrophysics Data System (ADS)

Oshika, Y.; Tsuchiya, Y.; Okumura, T.; Muramoto, Y.

2017-09-01

Ice has low environmental impact. Our research objectives are to study the availability of ice as a dielectric insulating material at cryogenic temperatures. We focus on ferroelectric ice (iceXI) at cryogenic temperatures. The properties of iceXI, including its formation, are not clear. We attempted to obtain the polarized ice that was similar to iceXI under the applied voltage and cooling to 77 K. The polarized ice have a wide range of engineering applications as electronic materials at cryogenic temperatures. This polarized ice is called ice electret. The structural difference between ice electret and normal ice is only the positions of protons. The effects of the proton arrangement on the breakdown voltage of ice electret were shown because electrical properties are influenced by the structure of ice. We observed an alternating current (ac) breakdown voltage of ice electret and normal ice at 77 K. The mean and minimum ac breakdown voltage values of ice electret were higher than those of normal ice. We considered that the electrically weak part of the normal ice was improved by applied a direct electric field.

Identification of new genes in a cell envelope-cell division gene cluster of Escherichia coli: cell envelope gene murG.

PubMed Central

Salmond, G P; Lutkenhaus, J F; Donachie, W D

1980-01-01

We report the identification, cloning, and mapping of a new cell envelope gene, murG. This lies in a group of five genes of similar phenotype (in the order murE murF murG murC ddl) all concerned with peptidoglycan biosynthesis. This group is in a larger cluster of at least 10 genes, all of which are involved in some way with cell envelope growth. Images PMID:6998962

Quantification of rare earth elements using laser-induced breakdown spectroscopy

DOE PAGES

Martin, Madhavi; Martin, Rodger C.; Allman, Steve; ...

2015-10-21

In this paper, a study of the optical emission as a function of concentration of laser-ablated yttrium (Y) and of six rare earth elements, europium (Eu), gadolinium (Gd), lanthanum (La), praseodymium (Pr), neodymium (Nd), and samarium (Sm), has been evaluated using the laser-induced breakdown spectroscopy (LIBS) technique. Statistical methodology using multivariate analysis has been used to obtain the sampling errors, coefficient of regression, calibration, and cross-validation of measurements as they relate to the LIBS analysis in graphite-matrix pellets that were doped with elements at several concentrations. Each element (in oxide form) was mixed in the graphite matrix in percentages rangingmore » from 1% to 50% by weight and the LIBS spectra obtained for each composition as well as for pure oxide samples. Finally, a single pellet was mixed with all the elements in equal oxide masses to determine if we can identify the elemental peaks in a mixed pellet. This dataset is relevant for future application to studies of fission product content and distribution in irradiated nuclear fuels. These results demonstrate that LIBS technique is inherently well suited for the future challenge of in situ analysis of nuclear materials. Finally, these studies also show that LIBS spectral analysis using statistical methodology can provide quantitative results and suggest an approach in future to the far more challenging multielemental analysis of ~ 20 primary elements in high-burnup nuclear reactor fuel.« less

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.

Distinct Perceptual Grouping Pathways Revealed By Temporal Carriers and Envelopes

PubMed Central

Rainville, Stéphane; Clarke, Aaron

2014-01-01

Guttman et al. [2005, Vis. Res., 45(8), 1021-1030] investigated whether observers could perform temporal grouping in multi-element displays where each local element was stochastically modulated over time along one of several potential dimensions – or “messenger types” – such as contrast, position, orientation, or spatial scale. Guttman et al.’s data revealed that grouping discards messenger type and therefore support a single-pathway model that groups elements with similar temporal waveforms. In the current study, we carried out three experiments in which temporal-grouping information resided either in the carrier, the envelope, or the combined carrier and envelope of each messenger’s timecourse. Results revealed that grouping is highly specific for messenger type if carrier envelopes lack grouping information but largely messenger nonspecific if carrier envelopes contain grouping information. The imply that temporal grouping is mediated by several messenger-specific carrier pathways as well as by a messenger-nonspecific envelope pathways. Findings also challenge simple temporal-filtering accounts of perceptual grouping [Adelson & Farid, 1999, Science, 286, 2231a]. PMID:19146293

Modification of selected South Carolina bridge-scour envelope curves

USGS Publications Warehouse

Benedict, Stephen T.; Caldwell, Andral W.

2012-01-01

Historic scour was investigated at 231 bridges in the Piedmont and Coastal Plain physiographic provinces of South Carolina by the U.S. Geological Survey in cooperation with the South Carolina Department of Transportation. These investigations led to the development of field-derived envelope curves that provided supplementary tools to assess the potential for scour at bridges in South Carolina for selected scour components that included clear-water abutment, contraction, and pier scour, and live-bed pier and contraction scour. The envelope curves consist of a single curve with one explanatory variable encompassing all of the measured field data for the respective scour components. In the current investigation, the clear-water abutment-scour and live-bed contraction-scour envelope curves were modified to include a family of curves that utilized two explanatory variables, providing a means to further refine the assessment of scour potential for those specific scour components. The modified envelope curves and guidance for their application are presented in this report.

Perfluorinated ionomer-enveloped sulfur cathodes for lithium-sulfur batteries.

PubMed

Song, Jongchan; Choo, Min-Ju; Noh, Hyungjun; Park, Jung-Ki; Kim, Hee-Tak

2014-12-01

Nafion is known to suppress the polysulfide (PS) shuttle effect, a major obstacle to achieving high capacity and long cycle life for lithium-sulfur batteries. However, elaborate control of the layer's configuration is required for high performance. In this regard, we designed a Nafion-enveloped sulfur cathode, where the Nafion layer is formed on the skin of the cathode, covering its surface and edge while not restricting the porosity. Discharge capacity and efficiency were enhanced with the enveloping configuration, demonstrating suppression of shuttle. The edge protection exhibited better cycling stability than an edge-open configuration. In the absence of the Nafion envelope, charged sulfur concentrated on the top region of the cathode because of the relatively lower PS concentration at the cathode surface. Surprisingly, for the Nafion-enveloped cathode, sulfur was evenly distributed along the cathode, indicating that the configuration imparts a uniform PS concentration within the cathode. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The nuclear import of ribosomal proteins is regulated by mTOR

PubMed Central

Kazyken, Dubek; Kaz, Yelimbek; Kiyan, Vladimir; Zhylkibayev, Assylbek A.; Chen, Chien-Hung; Agarwal, Nitin K.; Sarbassov, Dos D.

2014-01-01

Mechanistic target of rapamycin (mTOR) is a central component of the essential signaling pathway that regulates cell growth and proliferation by controlling anabolic processes in cells. mTOR exists in two distinct mTOR complexes known as mTORC1 and mTORC2 that reside mostly in cytoplasm. In our study, the biochemical characterization of mTOR led to discovery of its novel localization on nuclear envelope where it associates with a critical regulator of nuclear import Ran Binding Protein 2 (RanBP2). We show that association of mTOR with RanBP2 is dependent on the mTOR kinase activity that regulates the nuclear import of ribosomal proteins. The mTOR kinase inhibitors within thirty minutes caused a substantial decrease of ribosomal proteins in the nuclear but not cytoplasmic fraction. Detection of a nuclear accumulation of the GFP-tagged ribosomal protein rpL7a also indicated its dependence on the mTOR kinase activity. The nuclear abundance of ribosomal proteins was not affected by inhibition of mTOR Complex 1 (mTORC1) by rapamycin or deficiency of mTORC2, suggesting a distinctive role of the nuclear envelope mTOR complex in the nuclear import. Thus, we identified that mTOR in association with RanBP2 mediates the active nuclear import of ribosomal proteins. PMID:25294810

Visual Snapshots of Intracellular Kinase Activity At The Onset of Mitosis

PubMed Central

Dai, Zhaohua; Dulyaninova, Natalya G.; Kumar, Sanjai; Bresnick, Anne R.; Lawrence, David S.

2007-01-01

Summary Visual snapshots of intracellular kinase activity can be acquired with exquisite temporal control using a light-activatable (caged) sensor, thereby providing a means to interrogate enzymatic activity at any point during the cell division cycle. Robust protein kinase activity transpires just prior to, but not immediately following, nuclear envelope breakdown (NEB). Furthermore, kinase activity is required for progression from prophase into metaphase. Finally, the application of selective protein kinase C (PKC) inhibitors, in combination with the caged sensor, correlates the action of the PKC β isoform with subsequent NEB. PMID:18022564

Nuclear movement regulated by non-Smad Nodal signaling via JNK is associated with Smad signaling during zebrafish endoderm specification.

PubMed

Hozumi, Shunya; Aoki, Shun; Kikuchi, Yutaka

2017-11-01

Asymmetric nuclear positioning is observed during animal development, but its regulation and significance in cell differentiation remain poorly understood. Using zebrafish blastulae, we provide evidence that nuclear movement towards the yolk syncytial layer, which comprises extraembryonic tissue, occurs in the first cells fated to differentiate into the endoderm. Nodal signaling is essential for nuclear movement, whereas nuclear envelope proteins are involved in movement through microtubule formation. Positioning of the microtubule-organizing center, which is proposed to be crucial for nuclear movement, is regulated by Nodal signaling and nuclear envelope proteins. The non-Smad JNK signaling pathway, which is downstream of Nodal signaling, regulates nuclear movement independently of the Smad pathway, and this nuclear movement is associated with Smad signal transduction toward the nucleus. Our study provides insight into the function of nuclear movement in Smad signaling toward the nucleus, and could be applied to the control of TGFβ signaling. © 2017. Published by The Company of Biologists Ltd.

Breakdown in the organ donation process and its effect on organ availability.

PubMed

Razdan, Manik; Degenholtz, Howard B; Kahn, Jeremy M; Driessen, Julia

2015-01-01

Background. This study examines the effect of breakdown in the organ donation process on the availability of transplantable organs. A process breakdown is defined as a deviation from the organ donation protocol that may jeopardize organ recovery. Methods. A retrospective analysis of donation-eligible decedents was conducted using data from an independent organ procurement organization. Adjusted effect of process breakdown on organs transplanted from an eligible decedent was examined using multivariable zero-inflated Poisson regression. Results. An eligible decedent is four times more likely to become an organ donor when there is no process breakdown (adjusted OR: 4.01; 95% CI: 1.6838, 9.6414; P < 0.01) even after controlling for the decedent's age, gender, race, and whether or not a decedent had joined the state donor registry. However once the eligible decedent becomes a donor, whether or not there was a process breakdown does not affect the number of transplantable organs yielded. Overall, for every process breakdown occurring in the care of an eligible decedent, one less organ is available for transplant. Decedent's age is a strong predictor of likelihood of donation and the number of organs transplanted from a donor. Conclusion. Eliminating breakdowns in the donation process can potentially increase the number of organs available for transplant but some organs will still be lost.

Sensitivity to Envelope Interaural Time Differences at High Modulation Rates

PubMed Central

Bleeck, Stefan; McAlpine, David

2015-01-01

Sensitivity to interaural time differences (ITDs) conveyed in the temporal fine structure of low-frequency tones and the modulated envelopes of high-frequency sounds are considered comparable, particularly for envelopes shaped to transmit similar fidelity of temporal information normally present for low-frequency sounds. Nevertheless, discrimination performance for envelope modulation rates above a few hundred Hertz is reported to be poor—to the point of discrimination thresholds being unattainable—compared with the much higher (>1,000 Hz) limit for low-frequency ITD sensitivity, suggesting the presence of a low-pass filter in the envelope domain. Further, performance for identical modulation rates appears to decline with increasing carrier frequency, supporting the view that the low-pass characteristics observed for envelope ITD processing is carrier-frequency dependent. Here, we assessed listeners’ sensitivity to ITDs conveyed in pure tones and in the modulated envelopes of high-frequency tones. ITD discrimination for the modulated high-frequency tones was measured as a function of both modulation rate and carrier frequency. Some well-trained listeners appear able to discriminate ITDs extremely well, even at modulation rates well beyond 500 Hz, for 4-kHz carriers. For one listener, thresholds were even obtained for a modulation rate of 800 Hz. The highest modulation rate for which thresholds could be obtained declined with increasing carrier frequency for all listeners. At 10 kHz, the highest modulation rate at which thresholds could be obtained was 600 Hz. The upper limit of sensitivity to ITDs conveyed in the envelope of high-frequency modulated sounds appears to be higher than previously considered. PMID:26721926

Non-senescent Hydra tolerates severe disturbances in the nuclear lamina.

PubMed

Klimovich, Alexander; Rehm, Arvid; Wittlieb, Jörg; Herbst, Eva-Maria; Benavente, Ricardo; Bosch, Thomas C G

2018-05-10

The cnidarian Hydra is known for its unlimited lifespan and non-senescence, due to the indefinite self-renewal capacity of its stem cells. While proteins of the Lamin family are recognized as critical factors affecting senescence and longevity in human and mice, their putative role in the extreme longevity and non-senescence in long-living animals remains unknown. Here we analyze the role of a single lamin protein in non-senescence of Hydra . We demonstrate that proliferation of stem cells in Hydra is robust against the disturbance of Lamin expression and localization. While Lamin is indispensable for Hydra , the stem cells tolerate overexpression, downregulation and mislocalization of Lamin, and disturbances in the nuclear envelope structure. This extraordinary robustness may underlie the indefinite self-renewal capacity of stem cells and the non-senescence of Hydra . A relatively low complexity of the nuclear envelope architecture in basal Metazoa might allow for their extreme lifespans, while an increasing complexity of the nuclear architecture in bilaterians resulted in restricted lifespans.

A categorization of water system breakdowns: Evidence from Liberia, Nigeria, Tanzania, and Uganda.

PubMed

Klug, Tori; Cronk, Ryan; Shields, Katherine F; Bartram, Jamie

2018-04-01

In rural sub-Saharan Africa, one in three handpumps are non-functional at any time. While there is some evidence describing factors associated with non-functional water systems, there is little evidence describing the categories of water system breakdowns that commonly occur. Insufficient water availability from broken down systems can force people to use unimproved water sources, which undermines the health benefits of an improved water source. We categorized common water system breakdowns using quantitative and qualitative monitoring data from Liberia, Nigeria, Tanzania, and Uganda (each N>3600 water systems) and examined how breakdown category varies by water system type and management characteristics. Specific broken parts were mentioned more frequently than all other reasons for breakdown; hardware parts frequently found at fault for breakdown were aprons (Liberia), pipes (Tanzania and Uganda), taps/spouts (Tanzania and Uganda), and lift mechanisms (Nigeria). Statistically significant differences in breakdown category were identified based on system type, age, management type, and fee collection type. Categorization can help to identify common reasons for water system breakdown. The analysis of these data can be used to develop improved monitoring instruments to inform actors of different breakdown types and provide reasons for system non-functionality. Improved monitoring instruments would enable actors to target appropriate resources to address specific breakdowns likely to arise based on system type and management characteristics in order to inform improved implementation of and post-construction support for water systems in sub-Saharan Africa. Copyright © 2017 Elsevier B.V. All rights reserved.

Theoretical investigation of the breakdown electric field of SiC polymorphs

NASA Astrophysics Data System (ADS)

Yamaguchi, Kikou; Kobayashi, Daisuke; Yamamoto, Tomoyuki; Hirose, Kazuyuki

2018-03-01

The breakdown electric field of several SiC polymorphs has been investigated theoretically using a concept of "recovery rate," which is obtained by first principles calculations. A good relationship between the experimental breakdown electric fields and the calculated recovery rate of 4H-, 6H-, and 3C-SiC was obtained. In order to examine the stability of SiC polymorphs, the total electronic energies of various types of SiC crystal structures were calculated. Here, two candidates of polymorphs-GeS-type- and 2H-SiC-with energies comparable to those of experimentally well-established structures, have been obtained. The breakdown electric fields of these two polymorphs were estimated using a relationship obtained from the results of 4H-, 6H-, and 3C-SiC. This indicates that one of these polymorphs, GeS-type-SiC, has higher breakdown electric field than any other SiC polymorphs. In addition to the investigation with the recovery rate, relationship between experimental breakdown electric field and calculated band gap with recently developed accurate electron-correlation potential has been also discussed.

Breakdown characteristics of atmospheric dielectric barrier discharge in gas flow condition

NASA Astrophysics Data System (ADS)

Fan, Zhihui; Yan, Huijie; Wang, Yuying; Liu, Yidi; Guo, Hongfei; Ren, Chunsheng

2018-05-01

Experimental investigations of the breakdown characteristics of plate-to-plate dielectric barrier discharge excited by an AC source at different gas flow conditions are carried out. The ignition voltage for the appearance of the very first discharge filament and the breakdown voltage in each discharge half cycle in continuous operation are examined. As revealed by the results of the indoor air experiment, the ignition voltage manifests a monotonous increase with the increase in the gas flow rate, while the breakdown voltage has a marked decline at the low gas flow rate and increases slightly as the gas flow rate is higher than 10 m/s. As regards the obvious decreases in the ignition voltage and breakdown voltage, the decrease in the humidity with the increase in the gas flow rate plays a dominant role. As regards the increase in breakdown voltage, the memory effect from the preceding discharge is considered. The losses of metastable particles, together with particles having high translational energy in the gas flow, are considered to be the most critical factors.

Conservation of the egg envelope digestion mechanism of hatching enzyme in euteleostean fishes.

PubMed

Kawaguchi, Mari; Yasumasu, Shigeki; Shimizu, Akio; Sano, Kaori; Iuchi, Ichiro; Nishida, Mutsumi

2010-12-01

We purified two hatching enzymes, namely high choriolytic enzyme (HCE; EC 3.4.24.67) and low choriolytic enzyme (LCE; EC 3.4.24.66), from the hatching liquid of Fundulus heteroclitus, which were named Fundulus HCE (FHCE) and Fundulus LCE (FLCE). FHCE swelled the inner layer of egg envelope, and FLCE completely digested the FHCE-swollen envelope. In addition, we cloned three Fundulus cDNAs orthologous to cDNAs for the medaka precursors of egg envelope subunit proteins (i.e. choriogenins H, H minor and L) from the female liver. Cleavage sites of FHCE and FLCE on egg envelope subunit proteins were determined by comparing the N-terminal amino acid sequences of digests with the sequences deduced from the cDNAs for egg envelope subunit proteins. FHCE and FLCE cleaved different sites of the subunit proteins. FHCE efficiently cleaved the Pro-X-Y repeat regions into tripeptides to dodecapeptides to swell the envelope, whereas FLCE cleaved the inside of the zona pellucida domain, the core structure of egg envelope subunit protein, to completely digest the FHCE-swollen envelope. A comparison showed that the positions of hatching enzyme cleavage sites on egg envelope subunit proteins were strictly conserved between Fundulus and medaka. Finally, we extended such a comparison to three other euteleosts (i.e. three-spined stickleback, spotted halibut and rainbow trout) and found that the egg envelope digestion mechanism was well conserved among them. During evolution, the egg envelope digestion by HCE and LCE orthologs was established in the lineage of euteleosts, and the mechanism is suggested to be conserved. © 2010 The Authors Journal compilation © 2010 FEBS.

Carrier-envelope phase dynamics and noise analysis in octave-spanning Ti:sapphire lasers.

PubMed

Matos, Lia; Mücke, Oliver D; Chen, Jian; Kärtner, Franz X

2006-03-20

We investigate the carrier-envelope phase dynamics of octave-spanning Ti:sapphire lasers and perform a complete noise analysis of the carrier-envelope phase stabilization. We model the effect of the laser dynamics on the residual carrier-envelope phase noise by deriving a transfer function representation of the octave-spanning frequency comb. The modelled phase noise and the experimental results show excellent agreement. This greatly enhances our capability of predicting the dependence of the residual carrier-envelope phase noise on the feedback loop filter, the carrier-envelope frequency control mechanism and the pump laser used.

A statistical image analysis framework for pore-free islands derived from heterogeneity distribution of nuclear pore complexes.

PubMed

Mimura, Yasuhiro; Takemoto, Satoko; Tachibana, Taro; Ogawa, Yutaka; Nishimura, Masaomi; Yokota, Hideo; Imamoto, Naoko

2017-11-24

Nuclear pore complexes (NPCs) maintain cellular homeostasis by mediating nucleocytoplasmic transport. Although cyclin-dependent kinases (CDKs) regulate NPC assembly in interphase, the location of NPC assembly on the nuclear envelope is not clear. CDKs also regulate the disappearance of pore-free islands, which are nuclear envelope subdomains; this subdomain gradually disappears with increase in homogeneity of the NPC in response to CDK activity. However, a causal relationship between pore-free islands and NPC assembly remains unclear. Here, we elucidated mechanisms underlying NPC assembly from a new perspective by focusing on pore-free islands. We proposed a novel framework for image-based analysis to automatically determine the detailed 'landscape' of pore-free islands from a large quantity of images, leading to the identification of NPC intermediates that appear in pore-free islands with increased frequency in response to CDK activity. Comparison of the spatial distribution between simulated and the observed NPC intermediates within pore-free islands showed that their distribution was spatially biased. These results suggested that the disappearance of pore-free islands is highly related to de novo NPC assembly and indicated the existence of specific regulatory mechanisms for the spatial arrangement of NPC assembly on nuclear envelopes.

Evolution of Space Shuttle Range Safety Ascent Flight Envelope Design

NASA Technical Reports Server (NTRS)

Brewer, Joan; Davis, Jerel; Glenn, Christopher

2011-01-01

For every space vehicle launch from the Eastern Range in Florida, the range user must provide specific Range Safety (RS) data products to the Air Force's 45th Space Wing in order to obtain flight plan approval. One of these data products is a set of RS ascent flight envelope trajectories that define the normal operating region of the vehicle during powered flight. With the Shuttle Program launching 135 manned missions over a 30-year period, 135 envelope sets were delivered to the range. During this time, the envelope methodology and design process evolved to support mission changes, maintain high data quality, and reduce costs. The purpose of this document is to outline the shuttle envelope design evolution and capture the lessons learned that could apply to future spaceflight endeavors.

From organized high throughput data to phenomenological theory: The example of dielectric breakdown

NASA Astrophysics Data System (ADS)

Kim, Chiho; Pilania, Ghanshyam; Ramprasad, Rampi

Understanding the behavior (and failure) of dielectric insulators experiencing extreme electric fields is critical to the operation of present and emerging electrical and electronic devices. Despite its importance, the development of a predictive theory of dielectric breakdown has remained a challenge, owing to the complex multiscale nature of this process. Here, we focus on the intrinsic dielectric breakdown field of insulators--the theoretical limit of breakdown determined purely by the chemistry of the material, i.e., the elements the material is composed of, the atomic-level structure, and the bonding. Starting from a benchmark dataset (generated from laborious first principles computations) of the intrinsic dielectric breakdown field of a variety of model insulators, simple predictive phenomenological models of dielectric breakdown are distilled using advanced statistical or machine learning schemes, revealing key correlations and analytical relationships between the breakdown field and easily accessible material properties. The models are shown to be general, and can hence guide the screening and systematic identification of high electric field tolerant materials.

Feasibility Study of Endo- and Exo-skeletal Framed Structures with Envelopes for LTA Platforms

DTIC Science & Technology

2011-02-15

pathway for design and fabrication of Endo- and Exoskeleton framed elliptical envelopes was demonstrated. Envelope sizes of 2 ft x 0.5 ft and 5 ft x...Lighter than air, Endoskeleton, Exoskeleton , Helium filled envelope, Design, Fabrication Robert Sadler and Raghu Panduranga ARIS Inc 115-C, South...Structures with Envelopes for LTA Platforms Report Title ABSTRACT A pathway for design and fabrication of Endo- and Exoskeleton framed elliptical envelopes

Study of Bulk and Elementary Screw Dislocation Assisted Reverse Breakdown in Low-Voltage (< 250 V) 4H-SiC p(sup +)n Junction Diodes--Part II: Dynamic Breakdown Properties. Part 2; Dynamic Breakdown Properties

NASA Technical Reports Server (NTRS)

Neudeck, Philip G.; Fazi, Christian

1999-01-01

This paper outlines the dynamic reverse-breakdown characteristics of low-voltage (<250 V) small-area <5 x 10(exp -4) sq cm 4H-SiC p(sup +)n diodes subjected to nonadiabatic breakdown-bias pulsewidths ranging from 0.1 to 20 microseconds. 4H-SiC diodes with and without elementary screw dislocations exhibited positive temperature coefficient of breakdown voltage and high junction failure power densities approximately five times larger than the average failure power density of reliable silicon pn rectifiers. This result indicates that highly reliable low-voltage SiC rectifiers may be attainable despite the presence of elementary screw dislocations. However, the impact of elementary screw dislocations on other more useful 4H-SiC power device structures, such as high-voltage (>1 kV) pn junction and Schottky rectifiers, and bipolar gain devices (thyristors, IGBT's, etc.) remains to be investigated.

Carrier-envelope phase control by a composite plate.

PubMed

Ell, Richard; Birge, Jonathan R; Araghchini, Mohammad; Kärtner, Franz X

2006-06-12

We demonstrate a new concept to vary the carrier-envelope phase of a mode-locked laser by a composite plate while keeping all other pulse parameters practically unaltered. The effect is verified externally in an interferometric autocorrelator, as well as inside the cavity of an octave-spanning femtosecond oscillator. The carrier-envelope frequency can be shifted by half the repetition rate with negligible impact on pulse spectrum and energy.

Effects of polarization and absorption on laser induced optical breakdown threshold for skin rejuvenation

NASA Astrophysics Data System (ADS)

Varghese, Babu; Bonito, Valentina; Turco, Simona; Verhagen, Rieko

2016-03-01

Laser induced optical breakdown (LIOB) is a non-linear absorption process leading to plasma formation at locations where the threshold irradiance for breakdown is surpassed. In this paper we experimentally demonstrate the influence of polarization and absorption on laser induced breakdown threshold in transparent, absorbing and scattering phantoms made from water suspensions of polystyrene microspheres. We demonstrate that radially polarized light yields a lower irradiance threshold for creating optical breakdown compared to linearly polarized light. We also demonstrate that the thermal initiation pathway used for generating seed electrons results in a lower irradiance threshold compared to multiphoton initiation pathway used for optical breakdown.

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…

Scaling laws for AC gas breakdown and implications for universality

NASA Astrophysics Data System (ADS)

Loveless, Amanda M.; Garner, Allen L.

2017-10-01

The reduced dependence on secondary electron emission and electrode surface properties makes radiofrequency (RF) and microwave (MW) plasmas advantageous over direct current (DC) plasmas for various applications, such as microthrusters. Theoretical models relating molecular constants to alternating current (AC) breakdown often fail due to incomplete understanding of both the constants and the mechanisms involved. This work derives simple analytic expressions for RF and MW breakdown, demonstrating the transition between these regimes at their high and low frequency limits, respectively. We further show that the limiting expressions for DC, RF, and MW breakdown voltage all have the same universal scaling dependence on pressure and gap distance at high pressure, agreeing with experiment.

Evolution of centrosomes and the nuclear lamina: Amoebozoan assets.

PubMed

Gräf, Ralph; Batsios, Petros; Meyer, Irene

2015-06-01

The current eukaryotic tree of life groups most eukaryotes into one of five supergroups, the Opisthokonta, Amoebozoa, Archaeplastida, Excavata and SAR (Stramenopile, Alveolata, Rhizaria). Molecular and comparative morphological analyses revealed that the last eukaryotic common ancestor (LECA) already contained a rather sophisticated equipment of organelles including a mitochondrion, an endomembrane system, a nucleus with a lamina, a microtubule-organizing center (MTOC), and a flagellar apparatus. Recent studies of MTOCs, basal bodies/centrioles, and nuclear envelope organization of organisms in different supergroups have clarified our picture of how the nucleus and MTOCs co-evolved from LECA to extant eukaryotes. In this review we summarize these findings with special emphasis on valuable contributions of research on a lamin-like protein, nuclear envelope proteins, and the MTOC in the amoebozoan model organism Dictyostelium discoideum. Copyright © 2015 Elsevier GmbH. All rights reserved.

Two Improved Algorithms for Envelope and Wavefront Reduction

NASA Technical Reports Server (NTRS)

Kumfert, Gary; Pothen, Alex

1997-01-01

Two algorithms for reordering sparse, symmetric matrices or undirected graphs to reduce envelope and wavefront are considered. The first is a combinatorial algorithm introduced by Sloan and further developed by Duff, Reid, and Scott; we describe enhancements to the Sloan algorithm that improve its quality and reduce its run time. Our test problems fall into two classes with differing asymptotic behavior of their envelope parameters as a function of the weights in the Sloan algorithm. We describe an efficient 0(nlogn + m) time implementation of the Sloan algorithm, where n is the number of rows (vertices), and m is the number of nonzeros (edges). On a collection of test problems, the improved Sloan algorithm required, on the average, only twice the time required by the simpler Reverse Cuthill-Mckee algorithm while improving the mean square wavefront by a factor of three. The second algorithm is a hybrid that combines a spectral algorithm for envelope and wavefront reduction with a refinement step that uses a modified Sloan algorithm. The hybrid algorithm reduces the envelope size and mean square wavefront obtained from the Sloan algorithm at the cost of greater running times. We illustrate how these reductions translate into tangible benefits for frontal Cholesky factorization and incomplete factorization preconditioning.

Fast shut-down protection system for radio frequency breakdown and multipactor testing.

PubMed

Graves, T P; Hanson, P; Michaelson, J M; Farkas, A D; Hubble, A A

2014-02-01

Radio frequency (RF) breakdown such as multipactor or ionization breakdown is a device-limiting phenomenon for on-orbit spacecraft used for communication, navigation, or other RF payloads. Ground testing is therefore part of the qualification process for all high power components used in these space systems. This paper illustrates a shut-down protection system to be incorporated into multipactor/ionization breakdown ground testing for susceptible RF devices. This 8 channel system allows simultaneous use of different diagnostic classes and different noise floors. With initiation of a breakdown event, diagnostic signals increase above a user-specified level, which then opens an RF switch to eliminate RF power from the high power amplifier. Examples of this system in use are shown for a typical setup, illustrating the reproducibility of breakdown threshold voltages and the lack of multipactor conditioning. This system can also be utilized to prevent excessive damage to RF components in tests with sensitive or flight hardware.

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.

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.

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.

Fusion of Enveloped Viruses in Endosomes

PubMed Central

White, Judith M.; Whittaker, Gary R.

2016-01-01

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. PMID:26935856

High frequency vibration analysis by the complex envelope vectorization.

PubMed

Giannini, O; Carcaterra, A; Sestieri, A

2007-06-01

The complex envelope displacement analysis (CEDA) is a procedure to solve high frequency vibration and vibro-acoustic problems, providing the envelope of the physical solution. CEDA is based on a variable transformation mapping the high frequency oscillations into signals of low frequency content and has been successfully applied to one-dimensional systems. However, the extension to plates and vibro-acoustic fields met serious difficulties so that a general revision of the theory was carried out, leading finally to a new method, the complex envelope vectorization (CEV). In this paper the CEV method is described, underlying merits and limits of the procedure, and a set of applications to vibration and vibro-acoustic problems of increasing complexity are presented.

Prediction of breakdown strength of cellulosic insulating materials using artificial neural networks

NASA Astrophysics Data System (ADS)

Singh, Sakshi; Mohsin, M. M.; Masood, Aejaz

In this research work, a few sets of experiments have been performed in high voltage laboratory on various cellulosic insulating materials like diamond-dotted paper, paper phenolic sheets, cotton phenolic sheets, leatheroid, and presspaper, to measure different electrical parameters like breakdown strength, relative permittivity, loss tangent, etc. Considering the dependency of breakdown strength on other physical parameters, different Artificial Neural Network (ANN) models are proposed for the prediction of breakdown strength. The ANN model results are compared with those obtained experimentally and also with the values already predicted from an empirical relation suggested by Swanson and Dall. The reported results indicated that the breakdown strength predicted from the ANN model is in good agreement with the experimental values.

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.

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.

Spectral envelope sensitivity of musical instrument sounds.

PubMed

Gunawan, David; Sen, D

2008-01-01

It is well known that the spectral envelope is a perceptually salient attribute in musical instrument timbre perception. While a number of studies have explored discrimination thresholds for changes to the spectral envelope, the question of how sensitivity varies as a function of center frequency and bandwidth for musical instruments has yet to be addressed. In this paper a two-alternative forced-choice experiment was conducted to observe perceptual sensitivity to modifications made on trumpet, clarinet and viola sounds. The experiment involved attenuating 14 frequency bands for each instrument in order to determine discrimination thresholds as a function of center frequency and bandwidth. The results indicate that perceptual sensitivity is governed by the first few harmonics and sensitivity does not improve when extending the bandwidth any higher. However, sensitivity was found to decrease if changes were made only to the higher frequencies and continued to decrease as the distorted bandwidth was widened. The results are analyzed and discussed with respect to two other spectral envelope discrimination studies in the literature as well as what is predicted from a psychoacoustic model.

Mechanisms of nuclear lamina growth in interphase.

PubMed

Zhironkina, Oxana A; Kurchashova, Svetlana Yu; Pozharskaia, Vasilisa A; Cherepanynets, Varvara D; Strelkova, Olga S; Hozak, Pavel; Kireev, Igor I

2016-04-01

The nuclear lamina represents a multifunctional platform involved in such diverse yet interconnected processes as spatial organization of the genome, maintenance of mechanical stability of the nucleus, regulation of transcription and replication. Most of lamina activities are exerted through tethering of lamina-associated chromatin domains (LADs) to the nuclear periphery. Yet, the lamina is a dynamic structure demonstrating considerable expansion during the cell cycle to accommodate increased number of LADs formed during DNA replication. We analyzed dynamics of nuclear growth during interphase and changes in lamina structure as a function of cell cycle progression. The nuclear lamina demonstrates steady growth from G1 till G2, while quantitative analysis of lamina meshwork by super-resolution microscopy revealed that microdomain organization of the lamina is maintained, with lamin A and lamin B microdomain periodicity and interdomain gap sizes unchanged. FRAP analysis, in contrast, demonstrated differences in lamin A and B1 exchange rates; the latter showing higher recovery rate in S-phase cells. In order to further analyze the mechanism of lamina growth in interphase, we generated a lamina-free nuclear envelope in living interphase cells by reversible hypotonic shock. The nuclear envelope in nuclear buds formed after such a treatment initially lacked lamins, and analysis of lamina formation revealed striking difference in lamin A and B1 assembly: lamin A reassembled within 30 min post-treatment, whereas lamin B1 did not incorporate into the newly formed lamina at all. We suggest that in somatic cells lamin B1 meshwork growth is coordinated with replication of LADs, and lamin A meshwork assembly seems to be chromatin-independent process.

40 CFR 426.120 - Applicability; description of the incandescent lamp envelope manufacturing subcategory.

Code of Federal Regulations, 2013 CFR

2013-07-01

... incandescent lamp envelope manufacturing subcategory. 426.120 Section 426.120 Protection of Environment... POINT SOURCE CATEGORY Incandescent Lamp Envelope Manufacturing Subcategory § 426.120 Applicability; description of the incandescent lamp envelope manufacturing subcategory. The provisions of this subpart are...

40 CFR 426.120 - Applicability; description of the incandescent lamp envelope manufacturing subcategory.

Code of Federal Regulations, 2014 CFR

2014-07-01

... incandescent lamp envelope manufacturing subcategory. 426.120 Section 426.120 Protection of Environment... POINT SOURCE CATEGORY Incandescent Lamp Envelope Manufacturing Subcategory § 426.120 Applicability; description of the incandescent lamp envelope manufacturing subcategory. The provisions of this subpart are...

40 CFR 426.120 - Applicability; description of the incandescent lamp envelope manufacturing subcategory.

Code of Federal Regulations, 2012 CFR

2012-07-01

... incandescent lamp envelope manufacturing subcategory. 426.120 Section 426.120 Protection of Environment... POINT SOURCE CATEGORY Incandescent Lamp Envelope Manufacturing Subcategory § 426.120 Applicability; description of the incandescent lamp envelope manufacturing subcategory. The provisions of this subpart are...

40 CFR 426.120 - Applicability; description of the incandescent lamp envelope manufacturing subcategory.

Code of Federal Regulations, 2011 CFR

2011-07-01

... incandescent lamp envelope manufacturing subcategory. 426.120 Section 426.120 Protection of Environment... CATEGORY Incandescent Lamp Envelope Manufacturing Subcategory § 426.120 Applicability; description of the incandescent lamp envelope manufacturing subcategory. The provisions of this subpart are applicable to...

40 CFR 426.120 - Applicability; description of the incandescent lamp envelope manufacturing subcategory.

Code of Federal Regulations, 2010 CFR

2010-07-01

... incandescent lamp envelope manufacturing subcategory. 426.120 Section 426.120 Protection of Environment... CATEGORY Incandescent Lamp Envelope Manufacturing Subcategory § 426.120 Applicability; description of the incandescent lamp envelope manufacturing subcategory. The provisions of this subpart are applicable to...

Breakdown-Resistant RF Connectors for Vacuum

NASA Technical Reports Server (NTRS)

Caro, Edward R.; Bonazza, Walter J.

1987-01-01

Resilient inserts compensate for insulation shrinkage. Coaxial-cable connector for radio-frequency (RF) energy resists electrical breakdown in vacuum. Used on RF equipment in vacuum chambers as well as in spaceborne radar and communication gear.

Three Techniques for Task Analysis: Examples from the Nuclear Utilities.

ERIC Educational Resources Information Center

Carlisle, Kenneth E.

1984-01-01

Discusses three task analysis techniques utilized at the Palo Verde Nuclear Generating Station to review training programs: analysis of (1) job positions, (2) procedures, and (3) instructional presentations. All of these include task breakdown, relationship determination, and task restructuring. (MBR)

Improved model of activation energy absorption for different electrical breakdowns in semi-crystalline insulating polymers

NASA Astrophysics Data System (ADS)

Sima, Wenxia; Jiang, Xiongwei; Peng, Qingjun; Sun, Potao

2018-05-01

Electrical breakdown is an important physical phenomenon in electrical equipment and electronic devices. Many related models and theories of electrical breakdown have been proposed. However, a widely recognized understanding on the following phenomenon is still lacking: impulse breakdown strength which varies with waveform parameters, decrease in the breakdown strength of AC voltage with increasing frequency, and higher impulse breakdown strength than that of AC. In this work, an improved model of activation energy absorption for different electrical breakdowns in semi-crystalline insulating polymers is proposed based on the Harmonic oscillator model. Simulation and experimental results show that, the energy of trapped charges obtained from AC stress is higher than that of impulse voltage, and the absorbed activation energy increases with the increase in the electric field frequency. Meanwhile, the frequency-dependent relative dielectric constant ε r and dielectric loss tanδ also affect the absorption of activation energy. The absorbed activation energy and modified trap level synergistically determine the breakdown strength. The mechanism analysis of breakdown strength under various voltage waveforms is consistent with the experimental results. Therefore, the proposed model of activation energy absorption in the present work may provide a new possible method for analyzing and explaining the breakdown phenomenon in semi-crystalline insulating polymers.

Breakdown and invertebrate colonization of dead wood in wetland, upland, and river habitats

Treesearch

A Braccia; D Batzer

2010-01-01

Breakdown of woody debris in river and upland habitats as well as the interactions between wood and invertebrates have been well described. Studies of wood in wetlands are rare, and far less is known about breakdown and invertebrate use of wood in these transitional habitats. This study experimentally assessed breakdown and invertebrate colonization of wood in a...

Effects of elevated temperature on protein breakdown in muscles from septic rats

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hall-Angeras, M.A.; Angeras, U.H.; Hasselgren, P.O.

Elevated temperature has been proposed to contribute to accelerated muscle protein degradation during fever and sepsis. The present study examined the effect of increased temperature in vitro on protein turnover in skeletal muscles from septic and control rats. Sepsis was induced by cecal ligation and puncture (CLP); control rats were sham operated. After 16 h, the extensor digitorum longus (EDL) and soleus (SOL) muscles were incubated at 37 or 40 degrees C. Protein synthesis was determined by measuring incorporation of (14C)phenylalanine into protein. Total and myofibrillar protein breakdown was assessed from release of tyrosine and 3-methylhistidine (3-MH), respectively. Total proteinmore » breakdown was increased at 40 degrees C by 15% in EDL and by 29% in SOL from control rats, whereas 3-MH release was not affected. In muscles from septic rats, total and myofibrillar protein breakdown was increased by 22 and 30%, respectively, at 40 degrees C in EDL but was not altered in SOL. Protein synthesis was unaffected by high temperature both in septic and nonseptic muscles. The present results suggest that high temperature is not the primary mechanism of increased muscle protein breakdown in sepsis because the typical response to sepsis, i.e., a predominant increase in myofibrillar protein breakdown, was not induced by elevated temperature in normal muscle. It is possible, however, that increased temperature may potentiate protein breakdown that is already stimulated by sepsis because elevated temperature increased both total and myofibrillar protein breakdown in EDL from septic rats.« less

An experimental investigation of vortex breakdown on a delta wing

NASA Technical Reports Server (NTRS)

Payne, F. M.; Nelson, R. C.

1986-01-01

An experimental investigation of vortex breakdown on delta wings at high angles is presented. Thin delta wings having sweep angles of 70, 75, 80 and 85 degrees are being studied. Smoke flow visualization and the laser light sheet technique are being used to obtain cross-sectional views of the leading edge vortices as they break down. At low tunnel speeds (as low as 3 m/s) details of the flow, which are usually imperceptible or blurred at higher speeds, can be clearly seen. A combination of lateral and longitudinal cross-sectional views provides information on the three dimensional nature of the vortex structure before, during and after breakdown. Whereas details of the flow are identified in still photographs, the dynamic characteristics of the breakdown process were recorded using high speed movies. Velocity measurements were obtained using a laser Doppler anemometer with the 70 degree delta wing at 30 degrees angle of attack. The measurements show that when breakdown occurs the core flow transforms from a jet-like flow to a wake-like flow.

The Molecular and Dust Envelope of HD 56126

NASA Astrophysics Data System (ADS)

Meixner, M.; Zalucha, A.; Ueta, T.; Fong, D.; Justtanont, K.

2004-10-01

We present millimeter interferometry images of the CO J=1-0 line emission arising in the circumstellar envelope of HD 56126 (=IRAS 07134+1005), which is one of the best-studied 21 μm proto-planetary nebulae (PPNs). The CO emission extends from 1.2" to 7" in radius from the central star and appears consistent with a simple expanding envelope, as expected for a post-AGB star. The CO envelope is very clumpy with no apparent fast wind to explain these microstructures that must have arisen during the AGB mass loss. We quantitatively model the molecular envelope using a radiative transfer code that we have modified for detached shells. Our best-fit model reveals that two sequential winds created the circumstellar envelope of HD 56126: an AGB wind that lasted 6500 yr with a mass-loss rate of 5.1×10-6 Msolar yr-1 and a more intense superwind that lasted 840 yr with a mass-loss rate of 3×10-5 Msolar yr-1 and that ended the star's life on the AGB 1240 yr ago. The total mass of this envelope is 0.059 Msolar, which indicates a lower limit progenitor mass for the system of 0.66 Msolar, quite reasonable for this low-metallicity star that probably resides in the thick disk of the Galaxy. Comparison with images of the dust emission reveals a structure similar to that of the gas in the inner regions. Using 2-D UST, we model the dust emission of this source so that the model is consistent with the CO emission model and find a total dust mass of 7.8×10-4 Msolar, a superwind-dust mass-loss rate of 1.9×10-7 Msolar yr-1 and an AGB-dust mass-loss rate of 9.6×10-8 Msolar yr-1. We derive an average gas-to-dust mass ratio of 75, which is consistent with ISM values but low for what most consider for carbon stars. Our results indicate that TiC nanocrystals are probably not the carrier of the 21 μm feature.

Leaf breakdown in streams differing in catchment land use

USGS Publications Warehouse

Paul, M.J.; Meyer, J.L.; Couch, C.A.

2006-01-01

1. The impact of changes in land use on stream ecosystem function is poorly understood. We studied leaf breakdown, a fundamental process of stream ecosystems, in streams that represent a range of catchment land use in the Piedmont physiographic province of the south-eastern United States. 2. We placed bags of chalk maple (Acer barbatum) leaves in similar-sized streams in 12 catchments of differing dominant land use: four forested, three agricultural, two suburban and three urban catchments. We measured leaf mass, invertebrate abundance and fungal biomass in leaf bags over time. 3. Leaves decayed significantly faster in agricultural (0.0465 day-1) and urban (0.0474 day-1) streams than in suburban (0.0173 day-1) and forested (0.0100 day-1) streams. Additionally, breakdown rates in the agricultural and urban streams were among the fastest reported for deciduous leaves in any stream. Nutrient concentrations in agricultural streams were significantly higher than in any other land-use type. Fungal biomass associated with leaves was significantly lower in urban streams; while shredder abundance in leaf bags was significantly higher in forested and agricultural streams than in suburban and urban streams. Storm runoff was significantly higher in urban and suburban catchments that had higher impervious surface cover than forested or agricultural catchments. 4. We propose that processes accelerating leaf breakdown in agricultural and urban streams were not the same: faster breakdown in agricultural streams was due to increased biological activity as a result of nutrient enrichment, whereas faster breakdown in urban streams was a result of physical fragmentation resulting from higher storm runoff. ?? 2006 The Authors.

Surface Breakdown Characteristics of Silicone Oil for Electric Power Apparatus

NASA Astrophysics Data System (ADS)

Wada, Junichi; Nakajima, Akitoshi; Miyahara, Hideyuki; Takuma, Tadasu; Okabe, Shigemitu; Kohtoh, Masanori; Yanabu, Satoru

This paper describes the surface breakdown characteristics of the silicone oil which has the possibility of the application to innovative switchgear as an insulating medium. At the first step, we have experimentally studied on the impulse breakdown characteristics of the configuration with a triple-junction where a solid insulator is in contact with the electrode. The test configurations consist of solid material (Nomex and pressboard) and liquid insulation oil (silicone and mineral oil). We have discussed the experimental results based on the maximal electric field at a triple-junction. As the second step, we have studied the configuration which may improve the surface breakdown characteristics by lowering the electric field near the triple-junction.

Peculiarities of the circumstellar envelopes of some Herbig Ae/Be stars

NASA Astrophysics Data System (ADS)

Pogodin, M. A.

1985-10-01

The results of a spectral and photometric investigation of nine Herbig Ae/Be stars in the visible region of the spectrum in the period from 1978 to 1982 are presented. Certain physical and kinematic parameters of the circumstellar envelopes of the investigated objects are determined on the basis of the observational material obtained and Sobolev's (1947) probabilistic method for moving envelopes. It is shown that the observed spectral characteristics of the envelopes of Herbig Ae/Be stars and their variability can be explained by using models of extended isothermal envelopes with varying structural-kinematic parameters. The existence of a correlation between the amount of dust IR excess and the presence of signs of H2O absorption in the spectra of the investigated objects is noted.

Magnified Neural Envelope Coding Predicts Deficits in Speech Perception in Noise.

PubMed

Millman, Rebecca E; Mattys, Sven L; Gouws, André D; Prendergast, Garreth

2017-08-09

Verbal communication in noisy backgrounds is challenging. Understanding speech in background noise that fluctuates in intensity over time is particularly difficult for hearing-impaired listeners with a sensorineural hearing loss (SNHL). The reduction in fast-acting cochlear compression associated with SNHL exaggerates the perceived fluctuations in intensity in amplitude-modulated sounds. SNHL-induced changes in the coding of amplitude-modulated sounds may have a detrimental effect on the ability of SNHL listeners to understand speech in the presence of modulated background noise. To date, direct evidence for a link between magnified envelope coding and deficits in speech identification in modulated noise has been absent. Here, magnetoencephalography was used to quantify the effects of SNHL on phase locking to the temporal envelope of modulated noise (envelope coding) in human auditory cortex. Our results show that SNHL enhances the amplitude of envelope coding in posteromedial auditory cortex, whereas it enhances the fidelity of envelope coding in posteromedial and posterolateral auditory cortex. This dissociation was more evident in the right hemisphere, demonstrating functional lateralization in enhanced envelope coding in SNHL listeners. However, enhanced envelope coding was not perceptually beneficial. Our results also show that both hearing thresholds and, to a lesser extent, magnified cortical envelope coding in left posteromedial auditory cortex predict speech identification in modulated background noise. We propose a framework in which magnified envelope coding in posteromedial auditory cortex disrupts the segregation of speech from background noise, leading to deficits in speech perception in modulated background noise. SIGNIFICANCE STATEMENT People with hearing loss struggle to follow conversations in noisy environments. Background noise that fluctuates in intensity over time poses a particular challenge. Using magnetoencephalography, we demonstrate

Isolating The Building Thermal Envelope

NASA Astrophysics Data System (ADS)

Harrje, D. T.; Dutt, G. S.; Gadsby, K. J.

1981-01-01

The evaluation of the thermal integrity of building envelopes by infrared scanning tech-niques is often hampered in mild weather because temperature differentials across the envelope are small. Combining the infrared scanning with positive or negative building pressures, induced by a "blower door" or the building ventilation system, considerably extends the periods during which meaningful diagnostics can be conducted. Although missing or poorly installed insulation may lead to a substantial energy penalty, it is the search for air leakage sites that often has the largest potential for energy savings. Infrared inspection of the attic floor with air forced from the occupied space through ceiling by-passes, and inspecting the interior of the building when outside air is being sucked through the envelope reveals unexpected leakage sites. Portability of the diagnostic equipment is essential in these surveys which may include access into some tight spaces. A catalog of bypass heat losses that have been detected in residential housing using the combined infrared pressure differential technique is included to point out the wide variety of leakage sites which may compromise the benefits of thermal insulation and allow excessive air infiltration. Detection and suppression of such leaks should be key items in any building energy audit program. Where a calibrated blower door is used to pressurize or evacuate the house, the leakage rate can be quantified and an excessively tight house recognized. Houses that are too tight may be improved with a minimal energy penalty by forced ventilation,preferably with a heat recuperator and/or by providing combustion air directly to the furnace.

Method and apparatus for controlling carrier envelope phase

DOEpatents

Chang, Zenghu [Manhattan, KS; Li, Chengquan [Sunnyvale, CA; Moon, Eric [Manhattan, KS

2011-12-06

A chirped pulse amplification laser system. The system generally comprises a laser source, a pulse modification apparatus including first and second pulse modification elements separated by a separation distance, a positioning element, a measurement device, and a feedback controller. The laser source is operable to generate a laser pulse and the pulse modification apparatus operable to modify at least a portion of the laser pulse. The positioning element is operable to reposition at least a portion of the pulse modification apparatus to vary the separation distance. The measurement device is operable to measure the carrier envelope phase of the generated laser pulse and the feedback controller is operable to control the positioning element based on the measured carrier envelope phase to vary the separation distance of the pulse modification elements and control the carrier envelope phase of laser pulses generated by the laser source.

The nuclear lamina as a gene-silencing hub.

PubMed

Shevelyov, Yuri Y; Nurminsky, Dmitry I

2012-01-01

There is accumulating evidence that the nuclear periphery is a transcriptionally repressive compartment. A surprisingly large fraction of the genome is either in transient or permanent contact with nuclear envelope, where the majority of genes are maintained in a silent state, waiting to be awakened during cell differentiation. The integrity of the nuclear lamina and the histone deacetylase activity appear to be essential for gene repression at the nuclear periphery. However, the molecular mechanisms of silencing, as well as the events that lead to the activation of lamina-tethered genes, require further elucidation. This review summarizes recent advances in understanding of the mechanisms that link nuclear architecture, local chromatin structure, and gene regulation.

HIV-1 envelope glycoprotein

DOEpatents

Caulfield, Michael; Cupo, Albert; Dean, Hansi; Hoffenberg, Simon; King, C. Richter; Klasse, P. J.; Marozsan, Andre; Moore, John P.; Sanders, Rogier W.; Ward, Andrew; Wilson, Ian; Julien, Jean-Philippe

2017-08-22

The present application relates to novel HIV-1 envelope glycoproteins, which may be utilized as HIV-1 vaccine immunogens, and antigens for crystallization, electron microscopy and other biophysical, biochemical and immunological studies for the identification of broad neutralizing antibodies. The present invention encompasses the preparation and purification of immunogenic compositions, which are formulated into the vaccines of the present invention.

Quantum Hall effect breakdown in two-dimensional hole gases

NASA Astrophysics Data System (ADS)

Eaves, L.; Stoddart, S. T.; Wirtz, R.; Neumann, A. C.; Gallagher, B. L.; Main, P. C.; Henini, M.

2000-02-01

The breakdown of dissipationless current flow in the quantum Hall effect is studied for a two-dimensional hole gas at filling factors i=1 and 2. At high currents, the magnetoresistance curves at breakdown exhibit a series of steps accompanied by hysteresis and intermittent noise. These are compared with similar data for electron systems and are discussed in terms of a hydrodynamic model involving inter-Landau level scattering at the sample edge.

Thermal-envelope field measurements in an energy-efficient office/dormitory

DOE Office of Scientific and Technical Information (OSTI.GOV)

Christian, J.E.

1982-01-01

A 345 m/sup 2/ earth-covered structure located at the Oak Ridge National Laboratory is the focus of a DOE sponsored building-envelope research project. To heat the office/dormitory building over the 1981-1982 heating season would cost $1.70/m/sup 2/ ($0.16/ft/sup 2/), assuming $0.07/kWh. The thermal-integrity factor is 0.016 kWh/m/sup 2/ /sup 0/C (2.8 Btu/ft/sup 2/ /sup 0/F). A preliminary DOE-II model estimates the monthly electric energy needs for heating within 5% of field data derived estimates. DOE-II building simulations suggest that this earth-covered/passively heated office dormitory saves 30% for space heating and 26% for cooling compared to an energy efficient above grademore » structure. A preliminary winter energy balance has been generated from data collected in February and March providing a fractional breakdown of thermal losses and gains. A number of the energy-conserving component performances have been isolated; earth-covered roof, bermed wall, and nonvented trombe wall. The earth-covered roof system showed an overall thermal transmittance of 0.18 W/m/sup 2///sup 0/C (R=31 hr ft/sup 2/ /sup 0/F/Btu). The thermocouple wells located in the earth surrounding the building indicate the additional energy savings of burying over berming. For one week in February the trombe wall produced a 50% greater net thermal gain to the building then south facing windows per equivalent unit area.« less

Fast Positive Breakdown, NBEs, and Lightning Initiation

NASA Astrophysics Data System (ADS)

Krehbiel, P. R.; Rison, W.; Stock, M.; Edens, H. E.; Shao, X. M.; Thomas, R. J.; Stanley, M. A.; Zhang, Y.

2016-12-01

High power narrrow bipolar events (NBEs) have been found to be produced by arelatively unknown type of discharge, called fast positive breakdown (Rison etal., 2016). The breakdown occurs with a wide range of strengths, both in terms of its broadband sferic and its VHF radiation, and is found to be theinitiating event of many and likely all lightning discharges inside storms. Itdoes not produce a conducting channel but instead appears to be produced by avolumetric system of repeated, cascading positive streamers in virgin air.That positive corona and streamers would be responsible for initiatinglightning was proposed in the 1960s by Loeb, Dawson and Winn. In the 1970sPhelps and Griffiths showed that the streamers would be self-intensifying,leading to negative breakdown being initiated back at their starting points.Petersen et al. (2008) described experimental results showing that thestreamers could be initiated by ice crystals at cold temperatures, and thephysical processes leading to the breakdown being fast has been reported inrecent modeling studies by Shi et al. (2016). In this paper we summarize the observational data in support of the abovefindings, and report on additional observations of NBEs and lightninginitiation currently being obtained at Kennedy Space Center, Florida. References: Rison W., P.R. Krehbiel M.G.Stock, H.E. Edens, X-M. Shao, R.J. Thomas,M.A. Stanley, Y. Zhang, Observations of narrow bipolar events revealhow lightning is initiated in thunderstorms, Nature Comms. 7, 2016.doi:10.1038/ncomms10721. Petersen, D., Bailey, M., Beasley, W. & Hallett, J. A brief review ofthe problem of lightning initiation and a hypothesis of initiallightning leader formation. J. Geophys. Res. 113, D17205 (2008). Shi, F., N. Liu, and H. K. Rassoul (2016), Properties of relativelylong streamers initiated from an isolated hydrometeor, J. Geophys.Res. Atmos., 121, 7284-7295, doi:10.1002/2015JD024580.

Dredge-up and Envelope Burning in Intermediate-Mass Giants of Very Low Metallicity

NASA Astrophysics Data System (ADS)

Herwig, Falk

2004-04-01

The evolution of intermediate-mass stars at very low metallicity during their final thermal pulse asymptotic giant branch (AGB) phase is studied in detail. As representative examples, models with initial masses of 4 and 5Msolar and with a metallicity of Z=0.0001 ([Fe/H]~-2.3) are discussed. The one-dimensional stellar structure and evolution model includes time- and depth-dependent overshooting motivated by hydrodynamic simulations, as well as a full nuclear network and time-dependent mixing. Particular attention is given to high time and space resolution to avoid numerical artifacts related to third dredge-up and hot bottom burning predictions. The model calculations predict very efficient third dredge-up that mixes the envelope with the entire intershell layer or a large fraction thereof and in some cases penetrates into the C/O core below the He shell. In all cases primary oxygen is mixed into the envelope. The models predict efficient envelope burning during the interpulse phase. Depending on the envelope-burning temperature, oxygen is destroyed to varying degrees. The combined effect of dredge-up and envelope burning does not lead to any significant oxygen depletion in any of the cases considered in this study. The large dredge-up efficiency in our model is closely related to the particular properties of the H shell during the dredge-up phase in low-metallicity very metal-poor stars, which is followed here over many thermal pulses. During the dredge-up phase, the temperature just below the convective boundary is large enough for protons to burn vigorously when they are brought into the C-rich environment below the convection boundary by the time- and depth-dependent overshooting. H-burning luminosities of 105 to ~2×106Lsolar are generated. C, and to lesser degree O, is transformed into N in this dredge-up overshooting layer and enters the envelope. The global effect on the CNO abundance is similar to that of hot bottom burning. If the overshoot efficiency is

Intravenous Insulin Decreases Protein Breakdown in Infants on Extracorporeal Membrane Oxygenation

PubMed Central

Agus, Michael S.D.; Javid, Patrick J.; Ryan, Daniel P.; Jaksic, Tom

2010-01-01

Background/Purpose Infants requiring extracorporeal membrane oxygenation (ECMO) have the highest rates of protein catabolism ever reported. Recent investigations have found that such extreme protein breakdown is refractory to conventional nutritional management. In this pilot study, the authors sought to use the anabolic hormone insulin to reduce the profound protein degradation in this cohort. Methods Four parenterally fed infants on ECMO were enrolled in a prospective, randomized, crossover trial. Subjects were administered an insulin infusion using a 4-hour hyperinsulinemic euglycemic clamp followed by a control saline infusion on consecutive days in random order. Whole-body protein flux and breakdown were quantified using a primed continuous infusion of the stable isotope l-[1-13C]leucine. Statistical analyses were performed using paired t tests. Results Serum insulin levels were increased 15-fold during the insulin clamp compared with the saline control (407 ± 103 v 26 ± 12 µU/mL; P < .05). During the insulin infusion, infants had decreased rates of total leucine flux (214 ± 25 v 298 ± 38 µmol/kg/h; P < .05) and leucine flux derived from protein breakdown (156 ± 40 v 227 ± 54 µmol/kg/h; P < .05) when compared with saline control. Overall, insulin administration produced a 32% reduction in protein breakdown (P < .05). Conclusions In this pilot study, the anabolic hormone insulin markedly reduced protein breakdown in critically ill infants on ECMO. Because elevated protein breakdown correlates with mortality and morbidity, the administration of intravenous insulin may ultimately have broad applicability to the metabolic management of critically ill infants. PMID:15185208

Electric properties and carrier multiplication in breakdown sites in multi-crystalline silicon solar cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schneemann, Matthias; Carius, Reinhard; Rau, Uwe

2015-05-28

This paper studies the effective electrical size and carrier multiplication of breakdown sites in multi-crystalline silicon solar cells. The local series resistance limits the current of each breakdown site and is thereby linearizing the current-voltage characteristic. This fact allows the estimation of the effective electrical diameters to be as low as 100 nm. Using a laser beam induced current (LBIC) measurement with a high spatial resolution, we find carrier multiplication factors on the order of 30 (Zener-type breakdown) and 100 (avalanche breakdown) as new lower limits. Hence, we prove that also the so-called Zener-type breakdown is followed by avalanche multiplication. Wemore » explain that previous measurements of the carrier multiplication using thermography yield results higher than unity, only if the spatial defect density is high enough, and the illumination intensity is lower than what was used for the LBIC method. The individual series resistances of the breakdown sites limit the current through these breakdown sites. Therefore, the measured multiplication factors depend on the applied voltage as well as on the injected photocurrent. Both dependencies are successfully simulated using a series-resistance-limited diode model.« less

Identifying thermal breakdown products of thermoplastics.

PubMed

Guillemot, Marianne; Oury, Benoît; Melin, Sandrine

2017-07-01

Polymers processed to produce plastic articles are subjected to temperatures between 150°C and 450°C or more during overheated processing and breakdowns. Heat-based processing of this nature can lead to emission of volatile organic compounds (VOCs) into the thermoplastic processing shop. In this study, laboratory experiments, qualitative and quantitative emissions measurement in thermoplastic factories were carried out. The first step was to identify the compounds released depending on the thermoplastic nature, the temperature and the type of process. Then a thermal degradation protocol that can extrapolate the laboratory results to industry scenarios was developed. The influence of three parameters on released thermal breakdown products was studied: the sample preparation methods-manual cutting, ambient, or cold grinding-the heating rate during thermal degradation-5, 10 20, and 50°C/min-and the decomposition method-thermogravimetric analysis and pyrolysis. Laboratory results were compared to atmospheric measurements taken at 13 companies to validate the protocol and thereby ensure its representativeness of industrial thermal processing. This protocol was applied to most commonly used thermoplastics to determine their thermal breakdown products and their thermal behaviour. Emissions data collected by personal exposure monitoring and sampling at the process emission area show airborne concentrations of detected compounds to be in the range of 0-3 mg/m 3 under normal operating conditions. Laser cutting or purging operations generate higher pollution levels in particular formaldehyde which was found in some cases at a concentration above the workplace exposure limit.

The Red Queen in mitochondria: cyto-nuclear co-evolution, hybrid breakdown and human disease

PubMed Central

Chou, Jui-Yu; Leu, Jun-Yi

2015-01-01

Cyto-nuclear incompatibility, a specific form of Dobzhansky-Muller incompatibility caused by incompatible alleles between mitochondrial and nuclear genomes, has been suggested to play a critical role during speciation. Several features of the mitochondrial genome (mtDNA), including high mutation rate, dynamic genomic structure, and uniparental inheritance, make mtDNA more likely to accumulate mutations in the population. Once mtDNA has changed, the nuclear genome needs to play catch-up due to the intimate interactions between these two genomes. In two populations, if cyto-nuclear co-evolution is driven in different directions, it may eventually lead to hybrid incompatibility. Although cyto-nuclear incompatibility has been observed in a wide range of organisms, it remains unclear what type of mutations drives the co-evolution. Currently, evidence supporting adaptive mutations in mtDNA remains limited. On the other hand, it has been known that some mutations allow mtDNA to propagate more efficiently but compromise the host fitness (described as selfish mtDNA). Arms races between such selfish mtDNA and host nuclear genomes can accelerate cyto-nuclear co-evolution and lead to a phenomenon called the Red Queen Effect. Here, we discuss how the Red Queen Effect may contribute to the frequent observation of cyto-nuclear incompatibility and be the underlying driving force of some human mitochondrial diseases. PMID:26042149

On the breakdown modes and parameter space of Ohmic Tokamak startup

NASA Astrophysics Data System (ADS)

Peng, Yanli; Jiang, Wei; Zhang, Ya; Hu, Xiwei; Zhuang, Ge; Innocenti, Maria; Lapenta, Giovanni

2017-10-01

Tokamak plasma has to be hot. The process of turning the initial dilute neutral hydrogen gas at room temperature into fully ionized plasma is called tokamak startup. Even with over 40 years of research, the parameter ranges for the successful startup still aren't determined by numerical simulations but by trial and errors. However, in recent years it has drawn much attention due to one of the challenges faced by ITER: the maximum electric field for startup can't exceed 0.3 V/m, which makes the parameter range for successful startup narrower. Besides, this physical mechanism is far from being understood either theoretically or numerically. In this work, we have simulated the plasma breakdown phase driven by pure Ohmic heating using a particle-in-cell/Monte Carlo code, with the aim of giving a predictive parameter range for most tokamaks, even for ITER. We have found three situations during the discharge, as a function of the initial parameters: no breakdown, breakdown and runaway. Moreover, breakdown delay and volt-second consumption under different initial conditions are evaluated. In addition, we have simulated breakdown on ITER and confirmed that when the electric field is 0.3 V/m, the optimal pre-filling pressure is 0.001 Pa, which is in good agreement with ITER's design.

Effect of laser pulse repetition frequency on the optical breakdown threshold of quartz glass

NASA Astrophysics Data System (ADS)

Kononenko, T. V.; Schöneseiffen, S.; Konov, V. I.; Dausinger, F.

2013-08-01

The thresholds of optical breakdown in the volume of quartz glass were measured in relation to the number of pulses under irradiation by ultrashort laser pulses with different pulse repetition frequencies (1 - 400 kHz). Increasing this frequency from 10 to 400 kHz was found to substantially lower the breakdown threshold for 500-fs long pulses (at a wavelength of 1030 nm) and to lower to a smaller degree for 5-ps long pulses (515 nm). A strong frequency dependence of the breakdown threshold is observed under the same conditions as a manifold decrease of the breakdown threshold with increase in the number of pulses in a pulse train. The dependence of the optical breakdown on the number of pulses is attributable to the accumulation of point defects under multiple subthreshold irradiation, which affects the mechanism of collisional ionisation. In this case, the frequency dependence of the breakdown threshold of quartz glass is determined by the engagement of shortlived defects in the ionisation mechanism.

Effect of anode material on the breakdown in low-pressure helium gas

NASA Astrophysics Data System (ADS)

Demidov, V. I.; Adams, S. F.; Kudryavtsev, A. A.; Kurlyandskaya, I. P.; Miles, J. A.; Tolson, B. A.

2017-10-01

The electric breakdown of gases is one of the fundamental phenomena of gas discharge physics. It has been studied for a long time but still attracts incessant interest of researchers. Besides the interesting physics, breakdown is important for many applications including development of reliable electric insulation in electric grids and the study of different aspects of gas discharge physics. In this work an experimental study of the electric breakdown in helium gas for the plane-parallel electrode configuration has been conducted using a copper cathode and a variety of anode materials: copper, aluminum, stainless steel, graphite, platinum-plated aluminum and gold-plated aluminum. According to the Paschen law for studied electrode configuration, the breakdown voltage is a function of the product of gas pressure and inter-electrode gap. The breakdown processes on the left, lower pressure side of the Paschen curve have been the subject of this investigation. For those pressures, the Paschen curve may become multi-valued, where any given pressure corresponds to three breakdown voltage values. It was experimentally demonstrated that the form of the Paschen curve might strongly depend on the material of the anode and the cleanness of the anode surface. A possible explanation for this phenomenon is that electrons streaming from the cathode are reflected by the surface of the anode.

Structure and Chemical Composition of Prospheroplast Envelopes of Saccharomyces cerevisiae and Hansenula anomala

PubMed Central

Darling, Sven; Theilade, Jørgen; Birch-Andersen, Aksel

1972-01-01

Cells of Saccharomyces cerevisiae and Hansenula anomala were digested with snail enzyme under conditions yielding prospheroplasts. Surrounding envelopes were isolated after lysis of prospheroplasts in distilled water. The envelope material was embedded and sectioned for electron microscopy, and thin, hollow structures still retaining the elongated form of the original cells were seen. The envelopes were of low electron density in sections stained with uranyl magnesium acetate and lead citrate, but were more electron-dense when stained with phosphotungstic acid. Shadowed preparations of prospheroplast envelopes revealed structures resembling ghosts. These “ghosts” were similar to the original cells in form and size but seemed to be very thin. Varying numbers of anular structures (bud scars) were found on them. Chemical analyses of the envelope indicated that an alkali-soluble glucan was a major constituent. The results show that the prospheroplast envelope is part of the original cell wall of the yeast and is located in close apposition to the cytoplasmic membrane. Images PMID:4552997

Synthesis and transfer of galactolipids in the chloroplast envelope membranes of Arabidopsis thaliana.

PubMed

Kelly, Amélie A; Kalisch, Barbara; Hölzl, Georg; Schulze, Sandra; Thiele, Juliane; Melzer, Michael; Roston, Rebecca L; Benning, Christoph; Dörmann, Peter

2016-09-20

Galactolipids [monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG)] are the hallmark lipids of photosynthetic membranes. The galactolipid synthases MGD1 and DGD1 catalyze consecutive galactosyltransfer reactions but localize to the inner and outer chloroplast envelopes, respectively, necessitating intermembrane lipid transfer. Here we show that the N-terminal sequence of DGD1 (NDGD1) is required for galactolipid transfer between the envelopes. Different diglycosyllipid synthases (DGD1, DGD2, and Chloroflexus glucosyltransferase) were introduced into the dgd1-1 mutant of Arabidopsis in fusion with N-terminal extensions (NDGD1 and NDGD2) targeting to the outer envelope. Reconstruction of DGDG synthesis in the outer envelope membrane was observed only with diglycosyllipid synthase fusion proteins carrying NDGD1, indicating that NDGD1 enables galactolipid translocation between envelopes. NDGD1 binds to phosphatidic acid (PA) in membranes and mediates PA-dependent membrane fusion in vitro. These findings provide a mechanism for the sorting and selective channeling of lipid precursors between the galactolipid pools of the two envelope membranes.

Dynamic push-pull characteristics at three hand-reach envelopes: applications for the workplace.

PubMed

Calé-Benzoor, Maya; Dickstein, Ruth; Arnon, Michal; Ayalon, Moshe

2016-01-01

Pushing and pulling are common tasks in the workplace. Overexertion injuries related to manual pushing and pulling are often observed, and therefore the understanding of work capacity is important for efficient and safe workstation design. The purpose of the present study was to describe workloads obtained during different reach envelopes during a seated push-pull task. Forty-five women performed an isokinetic push-pull sequence at two velocities. Strength, work and agonist/antagonist muscle ratio were calculated for the full range of motion (ROM). We then divided the ROM into three reach envelopes - neutral, medium, and maximum reach. The work capacity for each direction was determined and the reach envelope work data were compared. Push capability was best at medium reach envelope and pulling was best at maximum reach envelope. Push/pull strength ratio was approximately 1. A recommendation was made to avoid strenuous push-pull tasks at neutral reach envelopes. Copyright © 2015 Elsevier Ltd and The Ergonomics Society. All rights reserved.

Time Dependent Dielectric Breakdown in Copper Low-k Interconnects: Mechanisms and Reliability Models

PubMed Central

Wong, Terence K.S.

2012-01-01

The time dependent dielectric breakdown phenomenon in copper low-k damascene interconnects for ultra large-scale integration is reviewed. The loss of insulation between neighboring interconnects represents an emerging back end-of-the-line reliability issue that is not fully understood. After describing the main dielectric leakage mechanisms in low-k materials (Poole-Frenkel and Schottky emission), the major dielectric reliability models that had appeared in the literature are discussed, namely: the Lloyd model, 1/E model, thermochemical E model, E1/2 models, E2 model and the Haase model. These models can be broadly categorized into those that consider only intrinsic breakdown (Lloyd, 1/E, E and Haase) and those that take into account copper migration in low-k materials (E1/2, E2). For each model, the physical assumptions and the proposed breakdown mechanism will be discussed, together with the quantitative relationship predicting the time to breakdown and supporting experimental data. Experimental attempts on validation of dielectric reliability models using data obtained from low field stressing are briefly discussed. The phenomenon of soft breakdown, which often precedes hard breakdown in porous ultra low-k materials, is highlighted for future research.