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Sample records for active nuclear import

  1. The nucleotide-binding domain of NLRC5 is critical for nuclear import and transactivation activity

    SciTech Connect

    Meissner, Torsten B.; Li, Amy; Liu, Yuen-Joyce; Gagnon, Etienne; Kobayashi, Koichi S.

    2012-02-24

    Highlights: Black-Right-Pointing-Pointer NLRC5 requires an intact NLS for its function as MHC class I transactivator. Black-Right-Pointing-Pointer Nuclear presence of NLRC5 is required for MHC class I induction. Black-Right-Pointing-Pointer Nucleotide-binding controls nuclear import and transactivation activity of NLRC5. -- Abstract: Major histocompatibility complex (MHC) class I and class II are crucial for the function of the human adaptive immune system. A member of the NLR (nucleotide-binding domain, leucine-rich repeat) protein family, NLRC5, has recently been identified as a transcriptional regulator of MHC class I and related genes. While a 'master regulator' of MHC class II genes, CIITA, has long been known, NLRC5 specifically associates with and transactivates the proximal promoters of MHC class I genes. In this study, we analyzed the molecular requirements of NLRC5 nuclear import and transactivation activity. We show that NLRC5-mediated MHC class I gene induction requires an intact nuclear localization signal and nuclear distribution of NLRC5. In addition, we find that the nucleotide-binding domain (NBD) of NLRC5 is critical not only for nuclear translocation but also for the transactivation of MHC class I genes. Changing the cellular localization of NLRC5 is likely to immediately impact MHC class I expression as well as MHC class I-mediated antigen presentation. NLRC5 may thus provide a promising target for the modulation of MHC class I antigen presentation, especially in the setting of transplant medicine.

  2. Identification of nuclear import and export signals within Fli-1: roles of the nuclear import signals in Fli-1-dependent activation of megakaryocyte-specific promoters.

    PubMed

    Hu, Wei; Philips, Alana S; Kwok, Juliana C; Eisbacher, Michael; Chong, Beng H

    2005-04-01

    The Ets factor Friend leukemia integration 1 (Fli-1) is an important regulator of megakaryocytic (Mk) differentiation. Here, we demonstrate two novel nuclear localization signals (NLSs) within Fli-1: one (NLS1) is located at the N terminus, and another (NLS2) is within the Ets domain. Nuclear accumulation of Fli-1 reflected the combined functional effects of the two discrete NLSs. Each NLS can independently direct nuclear transport of a carrier protein, with mutations within the NLSs affecting nuclear accumulation. NLS1 has a bipartite motif, whereas the NLS2 region contains a nonclassical NLS. Both NLSs bind importin alpha (IMPalpha) and IMPbeta, with NLS1 and NLS2 being predominantly recognized by IMPalpha and IMPbeta, respectively. Fli-1 also contains one nuclear export signal. Leptomycin B abolished its cytoplasmic accumulation, showing CRM1 dependency. We demonstrate that Ets domain binding to specific target DNA effectively blocks IMP binding, indicating that the targeted DNA binding plays a role in localizing Fli-1 to its destination and releasing IMPs for recycling back to the cytoplasm. Finally, by analyzing full-length Fli-1 carrying NLS1, NLS2, and combined NLS1-NLS2 mutations, we conclude that two functional NLSs exist in Fli-1 and that each NLS is sufficient to target Fli-1 to the nucleus for activation of Mk-specific genes.

  3. Acute β-Adrenergic Activation Triggers Nuclear Import of Histone Deacetylase 5 and Delays Gq-induced Transcriptional Activation*

    PubMed Central

    Chang, Chia-Wei Jenny; Lee, Linda; Yu, David; Dao, Khanha; Bossuyt, Julie; Bers, Donald M.

    2013-01-01

    During hemodynamic stress, catecholamines and neurohumoral stimuli may induce co-activation of Gq-coupled receptors and β-adrenergic receptors (β-AR), leading to cardiac remodeling. Dynamic regulation of histone deacetylase 5 (HDAC5), a transcriptional repressor, is crucial during stress signaling due to its role in epigenetic control of fetal gene markers. Little is known about its regulation during acute and chronic β-AR stimulation and its cross-interaction with Gq signaling in adult cardiac myocytes. Here, we evaluate the potential cross-talk between Gq-driven and β-AR mediated signaling at the level of nucleocytoplasmic shuttling of HDAC5. We show the translocation of GFP-tagged wild type HDAC5 or mutants (S279A and S279D) in response to β-AR or Gq agonists. Isoproterenol (ISO) or PKA activation results in strong nuclear accumulation of HDAC5 in contrast to nuclear export driven by Ca2+-calmodulin protein kinase II and protein kinase D. Moreover, nuclear accumulation of HDAC5 under acute ISO/PKA signaling is dependent on phosphorylation of Ser-279 and can block subsequent Gq-mediated nuclear HDAC5 export. Intriguingly, the attenuation of Gq-induced export is abolished after chronic PKA activation, yet nuclear HDAC5 remains elevated. Last, the effect of chronic β-AR signaling on HDAC5 translocation was examined in adult myocytes from a rabbit model of heart failure, where ISO-induced nuclear import is ablated, but Gq-agonist mediated export is preserved. Acute β-AR/PKA activation protects against hypertrophic signaling by delaying Gq-mediated transcriptional activation. This serves as a key physiological control switch before allowing genetic reprogramming via HDAC5 nuclear export during more severe stress, such as heart failure. PMID:23161540

  4. Isolation of CA1 nuclear enriched fractions from hippocampal slices to study activity-dependent nuclear import of synapto-nuclear messenger proteins.

    PubMed

    Yuanxiang, Pingan; Bera, Sujoy; Karpova, Anna; Kreutz, Michael R; Mikhaylova, Marina

    2014-08-10

    Studying activity dependent protein expression, subcellular translocation, or phosphorylation is essential to understand the underlying cellular mechanisms of synaptic plasticity. Long-term potentiation (LTP) and long-term depression (LTD) induced in acute hippocampal slices are widely accepted as cellular models of learning and memory. There are numerous studies that use live cell imaging or immunohistochemistry approaches to visualize activity dependent protein dynamics. However these methods rely on the suitability of antibodies for immunocytochemistry or overexpression of fluorescence-tagged proteins in single neurons. Immunoblotting of proteins is an alternative method providing independent confirmation of the findings. The first limiting factor in preparation of subcellular fractions from individual tetanized hippocampal slices is the low amount of material. Second, the handling procedure is crucial because even very short and minor manipulations of living slices might induce activation of certain signaling cascades. Here we describe an optimized workflow in order to obtain sufficient quantity of nuclear enriched fraction of sufficient purity from the CA1 region of acute hippocampal slices from rat brain. As a representative example we show that the ERK1/2 phosphorylated form of the synapto-nuclear protein messenger Jacob actively translocates to the nucleus upon induction of LTP and can be detected in a nuclear enriched fraction from CA1 neurons.

  5. Importance of nuclear localization for the apoptosis-induced activity of a fungal galectin AAL (Agrocybe aegerita lectin)

    SciTech Connect

    Liang, Yi; Feng, Lei; Tong, Xin; Wang, Kun; Li, De Feng; Lin, Jia Cheng; Tang, Zi Jian; Liu, Hong Hong; Jiang, Shuai; Guo, Lin; Wang, Da Cheng; Sun, Hui

    2009-08-28

    Agrocybe aegerita lectin (AAL) was identified previously in our group as a novel galectin from medicinal fungi Agrocybe aegerita, and has been shown to effectively induce cancer cell cycle arrest and apoptosis in vitro and tumor regression in vivo. Here, AAL was observed to translocate into the HeLa cell nucleus and induce cell apoptosis when it was predominantly in the nucleus. The N-terminus and C-terminus of AAL were required for nuclear localization. Site mutated proteins were generated based on AAL structure. Dimer interface mutant I25G, carbohydrate recognition domain (CRD) mutant R63H, and loop region mutant L33A could not enter the nucleus and lost the ability to induce apoptosis. CRD mutant H59Q and loop region mutant I144G maintained nuclear localization activity, and H59Q retained residual bioability but I144G had no activity, indicating that nuclear localization is important but not sufficient for AAL to become apoptotically active. Our findings provide a novel antitumor mechanism of fungal galectin.

  6. Sterol-dependent nuclear import of ORP1S promotes LXR regulated trans-activation of apoE

    SciTech Connect

    Lee, Sungsoo; Wang, Ping-Yuan; Jeong, Yangsik; Mangelsdorf, David J.; Anderson, Richard G.W.; Michaely, Peter

    2012-10-01

    Oxysterol binding protein related protein 1S (ORP1S) is a member of a family of sterol transport proteins. Here we present evidence that ORP1S translocates from the cytoplasm to the nucleus in response to sterol binding. The sterols that best promote nuclear import of ORP1S also activate the liver X receptor (LXR) transcription factors and we show that ORP1S binds to LXRs, promotes binding of LXRs to LXR response elements (LXREs) and specifically enhances LXR-dependent transcription via the ME.1 and ME.2 enhancer elements of the apoE gene. We propose that ORP1S is a cytoplasmic sterol sensor, which transports sterols to the nucleus and promotes LXR-dependent gene transcription through select enhancer elements. -- Highlights: Black-Right-Pointing-Pointer ORP1S translocates to the nucleus in response to sterol binding. Black-Right-Pointing-Pointer The sterols that best promote nuclear import of ORP1S are LXR agonists. Black-Right-Pointing-Pointer ORP1S binds to LXRs, enhances binding of LXRs to LXREs and promotes LXR-dependent transcription of apoE.

  7. Nuclear factors that bind two regions important to transcriptional activity of the simian immunodeficiency virus long terminal repeat.

    PubMed Central

    Winandy, S; Renjifo, B; Li, Y; Hopkins, N

    1992-01-01

    Previous studies identified two regions in the U3 region of a molecular clone of simian immunodeficiency virus, SIVmac142, that are important to transcriptional activity under conditions of induction as well as basal-level expression (B. Renjifo, N. A. Speck, S. Winandy, N. Hopkins, and Y. Li, J. Virol. 64:3130-3134, 1990). One region includes the NF-kappa B binding site, while the other lies just 5' of this site between nucleotides -162 and -114 (the -162 to -114 region). The fact that the NF-kappa B site mutation attenuated transcriptional activity in uninduced T cells and fibroblasts where activated NF-kappa B would not be present suggested that a factor(s) other than NF-kappa B could be acting through this site. In this study, we have identified a factor which binds to a cis element overlapping the NF-kappa B site. This factor, which we call simian factor 3 (SF3), would play a role in regulation under conditions of basal level expression, whereas under conditions of induction, NF-kappa B would act via this region. SF3 may also bind to an element in the -162 to -114 region. In addition, we have identified two other factors that bind the -162 to -114 region. One, which we designated SF1, is a ubiquitous basal factor, and the other, SF2, is a T-cell-predominant phorbol myristate acetate-inducible factor. Through identification of nuclear factors that interact with the U3 region of the SIVmac142 long terminal repeat, we can gain insight into how this virus is transcriptionally regulated under conditions of basal-level expression as well as conditions of T-cell activation. Images PMID:1501272

  8. Reconstitution of Nuclear Import in Permeabilized Cells

    PubMed Central

    Cassany, Aurelia; Gerace, Larry

    2012-01-01

    The trafficking of protein and RNA cargoes between the cytoplasm and the nucleus of eukaryotic cells, which is a major pathway involved in cell regulation, is mediated by nuclear transport sequences in the cargoes and by shuttling transport factors. The latter include receptors (karyopherins) that recognize the cargoes and carry them across the nuclear pore complex (NPC), and the small GTPase Ran, which modulates karyopherin–cargo binding. Nuclear import can be studied in vitro using digitonin-permeabilized cells, which are depleted of shuttling transport factors. Nuclear import can be reconstituted in the permeabilized cells with exogenous cytosol or with purified recombinant transport factors, and can be quantified by light microscopy of fluorescently labeled cargoes or by immunofluorescence staining. Here we describe procedures for in vitro nuclear import in permeabilized mammalian cells, and for the preparation of recombinant transport factors (importin α, importin β, importin 7, transportin, Ran, NTF2) and other reagents commonly used in the assay. This assay provides means to characterize the molecular mechanisms of nuclear import and to study the import requirements of specific cargoes. PMID:18951186

  9. Reconstitution of nuclear import in permeabilized cells.

    PubMed

    Cassany, Aurelia; Gerace, Larry

    2009-01-01

    The trafficking of protein and RNA cargoes between the cytoplasm and the nucleus of eukaryotic cells, which is a major pathway involved in cell regulation, is mediated by nuclear transport sequences in the cargoes and by shuttling transport factors. The latter include receptors (karyopherins) that recognize the cargoes and carry them across the nuclear pore complex (NPC), and the small GTPase Ran, which modulates karyopherin-cargo binding. Nuclear import can be studied in vitro using digitonin-permeabilized cells, which are depleted of shuttling transport factors. Nuclear import can be reconstituted in the permeabilized cells with exogenous cytosol or with purified recombinant transport factors, and can be quantified by light microscopy of fluorescently labeled cargoes or by immunofluorescence staining. Here we describe procedures for in vitro nuclear import in permeabilized mammalian cells, and for the preparation of recombinant transport factors (importin alpha, importin beta, importin 7, transportin, Ran, NTF2) and other reagents commonly used in the assay. This assay provides means to characterize the molecular mechanisms of nuclear import and to study the import requirements of specific cargoes.

  10. Ciliary Entry of the Hedgehog Transcriptional Activator Gli2 Is Mediated by the Nuclear Import Machinery but Differs from Nuclear Transport in Being Imp-α/β1-Independent.

    PubMed

    Torrado, Belén; Graña, Martín; Badano, José L; Irigoín, Florencia

    2016-01-01

    Gli2 is the primary transcriptional activator of Hedgehog signalling in mammals. Upon stimulation of the pathway, Gli2 moves into the cilium before reaching the nucleus. However, the mechanisms underlying its entry into the cilium are not completely understood. Since several similarities have been reported between nuclear and ciliary import, we investigated if the nuclear import machinery participates in Gli2 ciliary entry. Here we show that while two conserved classical nuclear localization signals mediate Gli2 nuclear localization via importin (Imp)-α/β1, these sequences are not required for Gli2 ciliary import. However, blocking Imp-mediated transport through overexpression of GTP-locked Ran reduced the percentage of Gli2 positive cilia, an effect that was not explained by increased CRM1-dependent export of Gli2 from the cilium. We explored the participation of Imp-β2 in Gli2 ciliary traffic and observed that this transporter is involved in moving Gli2 into the cilium, as has been described for other ciliary proteins. In addition, our data indicate that Imp-β2 might also collaborate in Gli2 nuclear entry. How does Imp-β2 determine the final destination of a protein that can localize to two distinct subcellular compartments remains an open question. Therefore, our data shows that the nuclear-cytoplasmic shuttling machinery plays a critical role mediating the subcellular distribution of Gli2 and the activation of the pathway, but distinct importins likely play a differential role mediating its ciliary and nuclear translocation.

  11. Ciliary Entry of the Hedgehog Transcriptional Activator Gli2 Is Mediated by the Nuclear Import Machinery but Differs from Nuclear Transport in Being Imp-α/β1-Independent

    PubMed Central

    Torrado, Belén; Graña, Martín; Badano, José L.; Irigoín, Florencia

    2016-01-01

    Gli2 is the primary transcriptional activator of Hedgehog signalling in mammals. Upon stimulation of the pathway, Gli2 moves into the cilium before reaching the nucleus. However, the mechanisms underlying its entry into the cilium are not completely understood. Since several similarities have been reported between nuclear and ciliary import, we investigated if the nuclear import machinery participates in Gli2 ciliary entry. Here we show that while two conserved classical nuclear localization signals mediate Gli2 nuclear localization via importin (Imp)-α/β1, these sequences are not required for Gli2 ciliary import. However, blocking Imp-mediated transport through overexpression of GTP-locked Ran reduced the percentage of Gli2 positive cilia, an effect that was not explained by increased CRM1-dependent export of Gli2 from the cilium. We explored the participation of Imp-β2 in Gli2 ciliary traffic and observed that this transporter is involved in moving Gli2 into the cilium, as has been described for other ciliary proteins. In addition, our data indicate that Imp-β2 might also collaborate in Gli2 nuclear entry. How does Imp-β2 determine the final destination of a protein that can localize to two distinct subcellular compartments remains an open question. Therefore, our data shows that the nuclear-cytoplasmic shuttling machinery plays a critical role mediating the subcellular distribution of Gli2 and the activation of the pathway, but distinct importins likely play a differential role mediating its ciliary and nuclear translocation. PMID:27579771

  12. PP2A Regulates HDAC4 Nuclear Import

    PubMed Central

    Paroni, Gabriela; Cernotta, Nadia; Dello Russo, Claudio; Gallinari, Paola; Pallaoro, Michele; Foti, Carmela; Talamo, Fabio; Orsatti, Laura; Steinkühler, Christian

    2008-01-01

    Different signal-regulated serine/threonine kinases phosphorylate class II histone deacetylases (HDACs) to promote nuclear export, cytosolic accumulation, and activation of gene transcription. However, little is known about mechanisms operating in the opposite direction, which, possibly through phosphatases, should promote class II HDACs nuclear entry and subsequent gene repression. Here we show that HDAC4 forms a complex with the PP2A holoenzyme Cα, Aα, B/PR55α. In vitro and in vivo binding studies demonstrate that the N-terminus of HDAC4 interacts with the catalytic subunit of PP2A. HDAC4 is dephosphorylated by PP2A and experiments using okadaic acid or RNA interference have revealed that PP2A controls HDAC4 nuclear import. Moreover, we identified serine 298 as a putative phosphorylation site important for HDAC4 nuclear import. The HDAC4 mutant mimicking phosphorylation of serine 298 is defective in nuclear import. Mutation of serine 298 to alanine partially rescues the defect in HDAC4 nuclear import observed in cells with down-regulated PP2A. These observations suggest that PP2A, via the dephosphorylation of multiple serines including the 14-3-3 binding sites and serine 298, controls HDAC4 nuclear import. PMID:18045992

  13. Monitoring international nuclear activity

    SciTech Connect

    Firestone, R.B.

    2006-05-19

    The LBNL Table of Isotopes website provides primary nuclearinformation to>150,000 different users annually. We have developedthe covert technology to identify users by IP address and country todetermine the kinds of nuclear information they are retrieving. Wepropose to develop pattern recognition software to provide an earlywarning system to identify Unusual nuclear activity by country or regionSpecific nuclear/radioactive material interests We have monitored nuclearinformation for over two years and provide this information to the FBIand LLNL. Intelligence is gleaned from the website log files. Thisproposal would expand our reporting capabilities.

  14. Computational identification of post-translational modification-based nuclear import regulations by characterizing nuclear localization signal-import receptor interaction.

    PubMed

    Lin, Jhih-Rong; Liu, Zhonghao; Hu, Jianjun

    2014-10-01

    The binding affinity between a nuclear localization signal (NLS) and its import receptor is closely related to corresponding nuclear import activity. PTM-based modulation of the NLS binding affinity to the import receptor is one of the most understood mechanisms to regulate nuclear import of proteins. However, identification of such regulation mechanisms is challenging due to the difficulty of assessing the impact of PTM on corresponding nuclear import activities. In this study we proposed NIpredict, an effective algorithm to predict nuclear import activity given its NLS, in which molecular interaction energy components (MIECs) were used to characterize the NLS-import receptor interaction, and the support vector regression machine (SVR) was used to learn the relationship between the characterized NLS-import receptor interaction and the corresponding nuclear import activity. Our experiments showed that nuclear import activity change due to NLS change could be accurately predicted by the NIpredict algorithm. Based on NIpredict, we developed a systematic framework to identify potential PTM-based nuclear import regulations for human and yeast nuclear proteins. Application of this approach has identified the potential nuclear import regulation mechanisms by phosphorylation of two nuclear proteins including SF1 and ORC6.

  15. Chikungunya virus capsid protein contains nuclear import and export signals

    PubMed Central

    2013-01-01

    Background Chikungunya virus (CHIKV) is an alphavirus of the Togaviridae family. After autoproteolytic cleavage, the CHIKV capsid protein (CP) is involved in RNA binding and assembly of the viral particle. The monomeric CP is approximately 30 kDa in size and is small enough for passive transport through nuclear pores. Some alphaviruses are found to harbor nuclear localization signals (NLS) and transport of these proteins between cellular compartments was shown to be energy dependent. The active nuclear import of cytoplasmic proteins is mediated by karyopherins and their export by exportins. As nuclear and cytoplasmic trafficking may play a role in the life cycle of CHIKV, we have sought to identify nuclear localization and nuclear export signals in CHIKV CP in a virus-free system. Methods EGFP-fusion proteins of CHIKV CP and mutants thereof were created and used to monitor their intracellular localization. Binding of cellular proteins was confirmed in pull-down assays with purified CP using co-immuoprecipitation. Nuclear localization was demonstrated in a virus-free system using fluorescence microscopy. Results Here we show that CHIKV CP is a nuclear-cytoplasmic shuttling protein with an active NLS that binds to karyopherin α (Karα) for its nuclear translocation. We also found that the Karα4 C-terminal NLS binding site is sufficient for this interaction. We further demonstrate that CHIKV CP interacts directly with the export receptor CRM1 to transport this viral protein out of the nucleus via a nuclear export signal (NES). The CHIKV CP NES was mapped between amino acids 143 and 155 of CP. Deduced from in silico analyses we found that the NES has a mode of binding similar to the snurportin-1 CRM1 complex. Conclusions We were able to show that in a virus-free system that the CHIKV capsid protein contains both, a NLS and a NES, and that it is actively transported between the cytoplasma and the nucleus. We conclude that CHIKV CP has the ability to shuttle via

  16. Blm10 facilitates nuclear import of proteasome core particles.

    PubMed

    Weberruss, Marion H; Savulescu, Anca F; Jando, Julia; Bissinger, Thomas; Harel, Amnon; Glickman, Michael H; Enenkel, Cordula

    2013-10-16

    Short-lived proteins are degraded by proteasome complexes, which contain a proteolytic core particle (CP) but differ in the number of regulatory particles (RPs) and activators. A recently described member of conserved proteasome activators is Blm10. Blm10 contains 32 HEAT-like modules and is structurally related to the nuclear import receptor importin/karyopherin β. In proliferating yeast, RP-CP assemblies are primarily nuclear and promote cell division. During quiescence, RP-CP assemblies dissociate and CP and RP are sequestered into motile cytosolic proteasome storage granuli (PSG). Here, we show that CP sequestration into PSG depends on Blm10, whereas RP sequestration into PSG is independent of Blm10. PSG rapidly clear upon the resumption of cell proliferation and proteasomes are relocated into the nucleus. Thereby, Blm10 facilitates nuclear import of CP. Blm10-bound CP serves as an import receptor-cargo complex, as Blm10 mediates the interaction with FG-rich nucleoporins and is dissociated from the CP by Ran-GTP. Thus, Blm10 represents the first CP-dedicated nuclear import receptor in yeast.

  17. Histone chaperones link histone nuclear import and chromatin assembly.

    PubMed

    Keck, Kristin M; Pemberton, Lucy F

    2013-01-01

    Histone chaperones are proteins that shield histones from nonspecific interactions until they are assembled into chromatin. After their synthesis in the cytoplasm, histones are bound by different histone chaperones, subjected to a series of posttranslational modifications and imported into the nucleus. These evolutionarily conserved modifications, including acetylation and methylation, can occur in the cytoplasm, but their role in regulating import is not well understood. As part of histone import complexes, histone chaperones may serve to protect the histones during transport, or they may be using histones to promote their own nuclear localization. In addition, there is evidence that histone chaperones can play an active role in the import of histones. Histone chaperones have also been shown to regulate the localization of important chromatin modifying enzymes. This review is focused on the role histone chaperones play in the early biogenesis of histones, the distinct cytoplasmic subcomplexes in which histone chaperones have been found in both yeast and mammalian cells and the importins/karyopherins and nuclear localization signals that mediate the nuclear import of histones. We also address the role that histone chaperone localization plays in human disease. This article is part of a Special Issue entitled: Histone chaperones and chromatin assembly.

  18. Multiple nuclear localization signals function in the nuclear import of the transcription factor Nrf2.

    PubMed

    Theodore, Melanie; Kawai, Yumiko; Yang, Jianqi; Kleshchenko, Yuliya; Reddy, Sekhar P; Villalta, Fernando; Arinze, Ifeanyi J

    2008-04-04

    Nuclear factor erythroid 2-related factor 2 (Nrf2) mediates the transcriptional response of cells to oxidative stress and is translocated into the nucleus following, or concomitant with, its activation by electrophiles or reactive oxygen species. The mechanism of its translocation into the nucleus is not entirely elucidated. Here we have identified two novel nuclear localization signal (NLS) motifs in murine Nrf2, one located near the N-terminal region (amino acid residues 42-53) and the other (residues 587-593) located near the C-terminal region. Imaging of green fluorescent protein (GFP)-tagged Nrf2 revealed that mutation(s) in any of these sequences resulted in decreased nuclear fluorescence intensity compared with the wild-type Nrf2 when Nrf2 activation was induced with the electrophile tert-butylhydroquinone. The mutations also impaired Nrf2-induced transactivation of antioxidant response element-driven reporter gene expression to the same extent as the Nrf2 construct bearing mutation in a previously identified bipartite NLS that maps at residues 494-511. When linked to GFP or to GFP-PEPCK-C each of the novel NLS motifs was sufficient to drive nuclear translocation of the fusion proteins. Co-immunoprecipitation assays demonstrated that importins alpha5 and beta1 associate with Nrf2, an interaction that was blocked by the nuclear import inhibitor SN50. SN50 also blocked tert-butylhydroquinone-induced nuclear fluorescence of GFP-Nrf2 in cells transfected with wild-type GFP-Nrf2. Overall these results reveal that multiple NLS motifs in Nrf2 function in its nuclear translocation in response to pro-oxidant stimuli and that the importin alpha-beta heterodimer nuclear import receptor system plays a critical role in the import process.

  19. Enhancing laboratory activities in nuclear medicine education.

    PubMed

    Grantham, Vesper; Martin, Chris; Schmitz, Casey

    2009-12-01

    Hands-on or active learning is important in nuclear medicine education. As more curricula start to require greater standards and as distance education expands, the effective use of laboratories in nuclear medicine education remains important in physics, instrumentation, and imaging but is often overlooked or underutilized. Laboratory exercises are a unique opportunity for nuclear medicine educators to facilitate students' critical thinking and problem-solving skills in a manner that often cannot occur in lectures or during online education. Given the lack of current laboratory tools and publications, there exists a requirement for nuclear medicine educators to develop, enhance, and monitor educational tools for laboratory exercises. Expanding technologies, variations in imaging and measurement systems, and the need to ensure that the taught technology is relevant to nuclear medicine students are issues faced by nuclear medicine educators. This article, based on principles of instructional design, focuses on the components and development of effective and enhanced nuclear medicine laboratories in our current educational environment.

  20. Importance of the regulation of nuclear receptor degradation.

    PubMed

    Dennis, A P; Haq, R U; Nawaz, Z

    2001-08-01

    Nuclear hormone receptors (NHRs) represent a superfamily of structurally related ligand-activated transcription factors, which regulate diverse biological activities like growth, development, and homeostasis. Recently, it has been demonstrated that certain members of the NHR superfamily are degraded through the ubiquitin-proteasome pathway in a ligand-dependent manner. Though the signal for the down-regulation via the ubiquitin-proteasome pathway is not yet known, phosphorylation at specific amino acid residues or coactivator binding to receptors could lead to their degradation by the 26S proteasome. Activation and degradation seems to be an engineered cyclic mechanism, which provides tight control over diverse cellular processes. The degradation process involves extensive loss of proteins and requires expenditure of cellular ATP. That seems to be inevitable for a more important aim, that is efficient and appropriate regulation of transcription. Down-regulation of receptors would lead to an attenuated transcriptional response because the number of receptor molecules available to activate transcription would decrease over time. One of the obvious reasons for down-regulating NHRs thus seems to be to prevent the cell from overstimulation by the hormones or other activating signals. Nuclear receptor turnover may also reset the transcriptional apparatus in preparation for a subsequent response. Since inhibition of the ubiquitin-proteasome degradation pathway disturbs the transcriptional activitity of some of the nuclear receptors such as estrogen (ER) and progesterone (PR) receptors, it is also possible that the degradation of NHRs may enable recycling of components of receptor-cofactor complexes and general transcriptional machinary. Understanding the mechanism of nuclear hormone receptor degradation and its relation to transcription may lead to novel insights of therapuetic intervention.

  1. Rapid nuclear import of short nucleic acids.

    PubMed

    Kitagawa, Mai; Okamoto, Akimitsu

    2016-10-01

    Exogenous short-chain nucleic acids undergo rapid import into the nucleus. Fluorescence-labeled dT1-13 DNA microinjected into the cytoplasm domain of a HeLa cell was rapidly imported into the nucleus domain within 1min. This is much more rapid than what has been observed for intracellular diffusion of small molecules. In contrast, import of longer nucleic acids with a length of over 30nt into the nucleus was suppressed.

  2. Intracellular calcium levels can regulate Importin-dependent nuclear import

    SciTech Connect

    Kaur, Gurpreet; Ly-Huynh, Jennifer D.; Jans, David A.

    2014-07-18

    Highlights: • High intracellular calcium inhibits Impα/β1- or Impβ1-dependent nuclear protein import. • The effect of Ca{sup 2+} on nuclear import does not relate to changes in the nuclear pore. • High intracellular calcium can result in mislocalisation of Impβ1, Ran and RCC1. - Abstract: We previously showed that increased intracellular calcium can modulate Importin (Imp)β1-dependent nuclear import of SRY-related chromatin remodeling proteins. Here we extend this work to show for the first time that high intracellular calcium inhibits Impα/β1- or Impβ1-dependent nuclear protein import generally. The basis of this relates to the mislocalisation of the transport factors Impβ1 and Ran, which show significantly higher nuclear localization in contrast to various other factors, and RCC1, which shows altered subnuclear localisation. The results here establish for the first time that intracellular calcium modulates conventional nuclear import through direct effects on the nuclear transport machinery.

  3. Teaching Activities on Horizontal Nuclear Proliferation.

    ERIC Educational Resources Information Center

    Zola, John

    1990-01-01

    Provides learning activities concerning the horizontal proliferation of nuclear weapons. Includes step-by-step directions for four activities: (1) the life cycle of nuclear weapons; (2) nuclear nonproliferation: pros and cons; (3) the nuclear power/nuclear weapons connection; and (4) managing nuclear proliferation. (NL)

  4. Importance of Nuclear Physics to NASA's Space Missions

    NASA Technical Reports Server (NTRS)

    Tripathi, R. K.; Wilson, J. W.; Cucinotta, F. A.

    2001-01-01

    We show that nuclear physics is extremely important for accurate risk assessments for space missions. Due to paucity of experimental input radiation interaction information it is imperative to develop reliable accurate models for the interaction of radiation with matter. State-of-the-art nuclear cross sections models have been developed at the NASA Langley Research center and are discussed.

  5. Nuclear protein import is reduced in cells expressing nuclear envelopathy-causing lamin A mutants

    SciTech Connect

    Busch, Albert; Kiel, Tilman; Heupel, Wolfgang-M.; Wehnert, Manfred; Huebner, Stefan

    2009-08-15

    Lamins, which form the nuclear lamina, not only constitute an important determinant of nuclear architecture, but additionally play essential roles in many nuclear functions. Mutations in A-type lamins cause a wide range of human genetic disorders (laminopathies). The importance of lamin A (LaA) in the spatial arrangement of nuclear pore complexes (NPCs) prompted us to study the role of LaA mutants in nuclear protein transport. Two mutants, causing prenatal skin disease restrictive dermopathy (RD) and the premature aging disease Hutchinson Gilford progeria syndrome, were used for expression in HeLa cells to investigate their impact on the subcellular localization of NPC-associated proteins and nuclear protein import. Furthermore, dynamics of the LaA mutants within the nuclear lamina were studied. We observed affected localization of NPC-associated proteins, diminished lamina dynamics for both LaA mutants and reduced nuclear import of representative cargo molecules. Intriguingly, both LaA mutants displayed similar effects on nuclear morphology and functions, despite their differences in disease severity. Reduced nuclear protein import was also seen in RD fibroblasts and impaired lamina dynamics for the nucleoporin Nup153. Our data thus represent the first study of a direct link between LaA mutant expression and reduced nuclear protein import.

  6. The importance of nuclear power in emissions avoidance

    SciTech Connect

    South, D.W.

    1999-11-01

    Most people knowledgeable about the nuclear power industry are aware that nuclear power plants do not emit air pollutants or greenhouse gases in the generation of electricity. What is commonly not known, however, is that these avoided emissions have become important for compliance with increasingly stringent limitations on air pollutants and greenhouse gas emissions required by the 1990 Clean Air Act Amendments and the Kyoto Protocol. This article is intended to heighten the awareness of this important environmental service and how its valuation can have an impact on the future operation of existing nuclear power plants

  7. Nuclear import of isoforms of the cytomegalovirus kinase pUL97 is mediated by differential activity of NLS1 and NLS2 both acting through classical importin-α binding.

    PubMed

    Webel, Rike; Solbak, Sara M Ø; Held, Christian; Milbradt, Jens; Groß, Andrea; Eichler, Jutta; Wittenberg, Thomas; Jardin, Christophe; Sticht, Heinrich; Fossen, Torgils; Marschall, Manfred

    2012-08-01

    The multifunctional protein kinase pUL97 of human cytomegalovirus (HCMV) strongly determines the efficiency of virus replication. Previously, the existence of two pUL97 isoforms that arise from alternative translational initiation and show a predominant nuclear localization was described. Two bipartite nuclear localization sequences, NLS1 and NLS2, were identified in the N terminus of the large isoform, whilst the small isoform exclusively contained NLS2. The current study found the following: (i) pUL97 nuclear localization in HCMV-infected primary fibroblasts showed accumulations in virus replication centres and other nuclear sections; (ii) in a quantitative evaluation system for NLS activity, the large isoform showed higher efficiency of nuclear translocation than the small isoform; (iii) NLS1 was mapped to aa 6-35 and NLS2 to aa 190-213; (iv) using surface plasmon resonance spectroscopy, the binding of both NLS1 and NLS2 to human importin-α was demonstrated, stressing the importance of individual arginine residues in the bipartite consensus motifs; (v) nuclear magnetic resonance spectroscopy of pUL97 peptides confirmed an earlier statement about the functional requirement of NLS1 embedding into an intact α-helical structure; and (vi) a bioinformatics investigation of the solvent-accessible surface suggested a high accessibility of NLS1 and an isoform-specific, variable accessibility of NLS2 for interaction with importin-α. Thus, the nucleocytoplasmic transport mechanism of the isoforms appeared to be differentially regulated, and this may have consequences for isoform-dependent functions of pUL97 during virus replication.

  8. New twist to nuclear import: When two travel together.

    PubMed

    Bange, Gert; Murat, Guillaume; Sinning, Irmgard; Hurt, Ed; Kressler, Dieter

    2013-07-01

    Ribosomes are the nanomachines that synthesize all cellular proteins from mRNA templates. In eukaryotes, ribosomes, which are composed of ribosomal proteins and rRNA, are mainly assembled in the nucleus. Thus, ribosomal proteins require a nuclear transport step from their place of synthesis in the cytoplasm to their site of assembly in the nucleus. Recognition of import substrates is mediated by different types of nuclear localization signals, which are either directly recognized by import receptors or recruited to these via adaptor proteins. The novel transport adaptor Syo1 (Symportin), which is dedicated to the synchronous import of two functionally related ribosomal proteins, has recently been described. In this review, we highlight and discuss these findings in the context of our current knowledge of ribosome assembly and nucleocytoplasmic transport. We propose that nuclear co-import of functionally and topologically linked cargo could be a widespread strategy to streamline assembly of macromolecular complexes in the nucleus.

  9. Importin α1 Mediates Yorkie Nuclear Import via an N-terminal Non-canonical Nuclear Localization Signal.

    PubMed

    Wang, Shimin; Lu, Yi; Yin, Meng-Xin; Wang, Chao; Wu, Wei; Li, Jinhui; Wu, Wenqing; Ge, Ling; Hu, Lianxin; Zhao, Yun; Zhang, Lei

    2016-04-08

    The Hippo signaling pathway controls organ size by orchestrating cell proliferation and apoptosis. When the Hippo pathway was inactivated, the transcriptional co-activator Yorkie translocates into the nucleus and forms a complex with transcription factor Scalloped to promote the expression of Hippo pathway target genes. Therefore, the nuclear translocation of Yorkie is a critical step in Hippo signaling. Here, we provide evidence that the N-terminal 1-55 amino acids of Yorkie, especially Arg-15, were essential for its nuclear localization. By mass spectrometry and biochemical analyses, we found that Importin α1 can directly interact with the Yorkie N terminus and drive Yorkie into the nucleus. Further experiments show that the upstream component Hippo can inhibit Importin α1-mediated Yorkie nuclear import. Taken together, we identified a potential nuclear localization signal at the N-terminal end of Yorkie as well as a critical role for Importin α1 in Yorkie nuclear import.

  10. Activation Experiments for Nuclear Astrophysics

    SciTech Connect

    Sonnabend, K.; Mueller, S.; Pietralla, N.; Savran, D.; Schnorrenberger, L.; Hasper, J.; Zilges, A.

    2009-01-28

    The study of ({gamma},n) reactions can be used to constrain the theoretical predictions of the neutron capture cross sections of short-lived branching points in the s process. The usability of the activation technique to study these ({gamma},n) reactions is discussed as one example of an activation experiment in nuclear astrophysics. Two photon sources using bremsstrahlung and laser-Compton backscattered photons where such experiments were carried out are compared.

  11. Importance of risk communication during and after a nuclear accident.

    PubMed

    Perko, Tanja

    2011-07-01

    Past nuclear accidents highlight communication as one of the most important challenges in emergency management. In the early phase, communication increases awareness and understanding of protective actions and improves the population response. In the medium and long term, risk communication can facilitate the remediation process and the return to normal life. Mass media play a central role in risk communication. The recent nuclear accident in Japan, as expected, induced massive media coverage. Media were employed to communicate with the public during the contamination phase, and they will play the same important role in the clean-up and recovery phases. However, media also have to fulfill the economic aspects of publishing or broadcasting, with the "bad news is good news" slogan that is a well-known phenomenon in journalism. This article addresses the main communication challenges and suggests possible risk communication approaches to adopt in the case of a nuclear accident.

  12. Multiple Conserved Domains of the Nucleoporin Nup124p and Its Orthologs Nup1p and Nup153 Are Critical for Nuclear Import and Activity of the Fission Yeast Tf1 Retrotransposon

    PubMed Central

    Sistla, Srivani; Pang, Junxiong Vincent; Wang, Cui Xia

    2007-01-01

    The nucleoporin Nup124p is a host protein required for the nuclear import of both, retrotransposon Tf1-Gag as well as the retroviral HIV-1 Vpr in fission yeast. The human nucleoporin Nup153 and the Saccharomyces cerevisiae Nup1p were identified as orthologs of Nup124p. In this study, we show that all three nucleoporins share a large FG/FXFG-repeat domain and a C-terminal peptide sequence, GRKIxxxxxRRKx, that are absolutely essential for Tf1 retrotransposition. Though the FXFG domain was essential, the FXFG repeats themselves could be eliminated without loss of retrotransposon activity, suggesting the existence of a common element unrelated to FG/FXFG motifs. The Nup124p C-terminal peptide, GRKIAVPRSRRKR, was extremely sensitive to certain single amino acid changes within stretches of the basic residues. On the basis of our comparative study of Nup124p, Nup1p, and Nup153 domains, we have developed peptides that specifically knockdown retrotransposon activity by disengaging the Tf1-Gag from its host nuclear transport machinery without any harmful consequence to the host itself. Our results imply that those domains challenged a specific pathway affecting Tf1 transposition. Although full-length Nup1p or Nup153 does not complement Nup124p, the functionality of their conserved domains with reference to Tf1 activity suggests that these three proteins evolved from a common ancestor. PMID:17615301

  13. Row erupts over US firm's plan to import nuclear waste

    NASA Astrophysics Data System (ADS)

    Gwynne, Peter

    2008-06-01

    A controversy is brewing in the US over a plan by a firm in Utah to import, process and dispose of 20 000 tonnes of low-level radioactive waste from decommissioned nuclear reactors built in Italy by American companies. EnergySolutions intends to recycle some of this waste at a site near Oak Ridge, Tennessee, so that it can be re-used as shielding blocks in nuclear plants. The firm then wants to dispose of the remaining radioactive material at a site in Clive, Utah, where over 90% of low-level radioactive waste generated in the US is currently buried.

  14. The Importance of International Technical Nuclear Forensics to Deter Illicit Trafficking

    SciTech Connect

    Smith, D K

    2007-01-30

    Illicit trafficking of nuclear materials is a transboundary problem that requires a cooperative approach involving international nuclear forensics to ensure all states understand the threat posed by nuclear smuggling as well as a means to best deter the movement of nuclear contraband. To achieve the objectives, all cases involving illicit trafficking of nuclear and radiological materials must be vigorously pursued and prosecuted when appropriate. The importance of outreach and formal government-to-government relationships with partner nations affected by nuclear trafficking cannot be under-estimated. States that are situated on smuggling routes may be well motivated to counter nuclear crimes to bolster their own border and transportation security as well as strengthen their economic and political viability. National law enforcement and atomic energy agencies in these states are aggressively pursuing a comprehensive strategy to counter nuclear smuggling through increasing reliance on technical nuclear forensics. As part of these activities, it is essential that these organizations be given adequate orientation to the best practices in this emerging discipline including the categorization of interdicted nuclear material, collection of traditional and nuclear forensic evidence, data analysis using optimized analytical protocols, and how to best fuse forensics information with reliable case input to best develop a law enforcement or national security response. The purpose of formalized USG relationship is to establish an institutional framework for collaboration in international forensics, improve standards of forensics practice, conduct joint exercises, and pursue case-work that benefits international security objectives. Just as outreach and formalized relationships are important to cultivate international nuclear forensics, linking nuclear forensics to ongoing national assistance in border and transpiration security, including port of entry of entry monitoring

  15. A photoregulated ligand for the nuclear import receptor karyopherin alpha.

    PubMed

    Park, S B; Standaert, R F

    2001-12-01

    The ability to orchestrate the transport of proteins between nucleus and cytoplasm provides cells with a powerful regulatory mechanism. Selective translocation between these compartments is often used to propagate cellular signals, and it is an intimate part of the processes that control cell division, viral replication, and other cellular events. Therefore, precise experimental control over protein localization, through the agency of light, would provide a powerful tool for the study and manipulation of these events. To this end, a prototype photoregulated nuclear localization signal (NLS) was derived from a native NLS. A library of 30 mutants of the bipartite NLS from Xenopus laevis nucleoplasmin containing a novel, photoisomerizable amino acid was prepared by parallel, solid-phase synthesis and screened in vitro for binding to the nuclear import receptor karyopherin alpha, which mediates the nuclear import of cellular proteins. A single peptide was identified in which the cis and trans photoisomers bind the receptor differentially. The strategy used to obtain this peptide is systematic and empirical; therefore, it is potentially applicable to any peptide-receptor system.

  16. Experimental Nuclear Physics Activity in Italy

    NASA Astrophysics Data System (ADS)

    Chiavassa, E.; de Marco, N.

    2003-04-01

    The experimental Nuclear Physics activity of the Italian researchers is briefly reviewed. The experiments, that are financially supported by the INFN, are done in strict collaboration by more than 500 INFN and University researchers. The experiments cover all the most important field of the modern Nuclear Physics with probes extremely different in energy and interactions. Researches are done in all the four National Laboratories of the INFN even if there is a deeper involvement of the two national laboratories expressly dedicated to Nuclear Physics: the LNL (Laboratorio Nazionale di Legnaro) and LNS (Laboratorio Nazionale del Sud) where nuclear spectroscopy and reaction dynamics are investigated. All the activities with electromagnetic probes develops in abroad laboratories as TJNAF, DESY, MAMI, ESFR and are dedicated to the studies of the spin physics and of the nucleon resonance; hypernuclear and kaon physics is investigated at LNF. A strong community of researchers work in the relativistic and ultra-relativistic heavy ions field in particular at CERN with the SPS Pb beam and in the construction of the ALICE detector for heavy-ion physics at the LHC collider. Experiments of astrophysical interest are done with ions of very low energy; in particular the LUNA accelerator facility at LNGS (Laboratorio Nazionale del Gran Sasso) succeeded measuring cross section at solar energies, below or near the solar Gamow peak. Interdisciplinary researches on anti-hydrogen atom spectroscopy and on measurements of neutron cross sections of interest for ADS development are also supported.

  17. Evolving Important Role of Lutetium-177 for Therapeutic Nuclear Medicine.

    PubMed

    Pillai, Ambikalmajan M R; Knapp, Furn F Russ

    2015-01-01

    Lutetium-177 ((177)Lu) is a late entrant into the nuclear medicine therapy arena but is expected to become one of the most widely used therapeutic radionuclides. This paper analyses the reason for the increasing preference of (177)Lu as a therapeutic radionuclide. While the radionuclidic properties favor its use for several therapeutic applications, the potential for large scale production of (177)Lu is also an important aspect for its acceptability as a therapeutic radionuclide. This introductory discussion also summarizes some developing clinical uses and suggested future directions for applications of (177)Lu.

  18. Karyopherin Alpha 1 Regulates Satellite Cell Proliferation and Survival by Modulating Nuclear Import.

    PubMed

    Choo, Hyo-Jung; Cutler, Alicia; Pavlath, Grace K

    2016-07-19

    Satellite cells are stem cells with an essential role in skeletal muscle repair. Precise regulation of gene expression is critical for proper satellite cell quiescence, proliferation, differentiation and self-renewal. Nuclear proteins required for gene expression are dependent on the nucleocytoplasmic transport machinery to access to nucleus, however little is known about regulation of nuclear transport in satellite cells. The best characterized nuclear import pathway is classical nuclear import which depends on a classical nuclear localization signal (cNLS) in a cargo protein and the heterodimeric import receptors, karyopherin alpha (KPNA) and beta (KPNB). Multiple KPNA1 paralogs exist and can differ in importing specific cNLS proteins required for cell differentiation and function. We show that transcripts for six Kpna paralogs underwent distinct changes in mouse satellite cells during muscle regeneration accompanied by changes in cNLS proteins in nuclei. Depletion of KPNA1, the most dramatically altered KPNA, caused satellite cells in uninjured muscle to prematurely activate, proliferate and undergo apoptosis leading to satellite cell exhaustion with age. Increased proliferation of satellite cells led to enhanced muscle regeneration at early stages of regeneration. In addition, we observed impaired nuclear localization of two key KPNA1 cargo proteins: p27, a cyclin-dependent kinase inhibitor associated with cell cycle control and lymphoid enhancer factor 1, a critical cotranscription factor for β-catenin. These results indicate that regulated nuclear import of proteins by KPNA1 is critical for satellite cell proliferation and survival and establish classical nuclear import as a novel regulatory mechanism for controlling satellite cell fate. Stem Cells 2016.

  19. Preventing the importation of illicit nuclear materials in shipping containers.

    PubMed

    Wein, Lawrence M; Wilkins, Alex H; Baveja, Manas; Flynn, Stephen E

    2006-10-01

    We develop a mathematical model to find the optimal inspection strategy for detecting a nuclear weapon (or nuclear material to make a weapon) from being smuggled into the United States in a shipping container, subject to constraints of port congestion and an overall budget. We consider an 11-layer security system consisting of shipper certification, container seals, and a targeting software system, followed by passive (neutron and gamma), active (gamma radiography), and manual testing at overseas and domestic ports. Currently implemented policies achieve a low detection probability, and improved security requires passive and active testing of trusted containers and manually opening containers that cannot be penetrated by radiography. The annual cost of achieving a high detection probability of a plutonium weapon using existing equipment in traditional ways is roughly several billion dollars if testing is done domestically, and is approximately five times higher if testing is performed overseas. Our results suggest that employing high-energy x-ray radiography and elongating the passive neutron tests at overseas ports may provide significant cost savings, and several developing technologies, radiation sensors inside containers and tamper-resistant electronic seals, should be pursued aggressively. Further effort is critically needed to develop a practical neutron interrogation scheme that reliably detects moderately shielded, highly enriched uranium.

  20. Nuclear localization signals for four distinct karyopherin-β nuclear import systems.

    PubMed

    Soniat, Michael; Chook, Yuh Min

    2015-06-15

    The Karyopherin-β family of proteins mediates nuclear transport of macromolecules. Nuclear versus cytoplasmic localization of proteins is often suggested by the presence of NLSs (nuclear localization signals) or NESs (nuclear export signals). Import-Karyopherin-βs or Importins bind to NLSs in their protein cargos to transport them through nuclear pore complexes into the nucleus. Until recently, only two classes of NLS had been biochemically and structurally characterized: the classical NLS, which is recognized by the Importin-α/β heterodimer and the PY-NLS (proline-tyrosine NLS), which is recognized by Karyopherin-β2 or Transportin-1. Structures of two other Karyopherin-βs, Kap121 and Transportin-SR2, in complex with their respective cargos were reported for the first time recently, revealing two new distinct classes of NLSs. The present paper briefly describes the classical NLS, reviews recent literature on the PY-NLS and provides in-depth reviews of the two newly discovered classes of NLSs that bind Kap121p and Transportin-SR respectively.

  1. Building Public Confidence in Nuclear Activities

    SciTech Connect

    Isaacs, T

    2002-02-13

    Achieving public acceptance has become a central issue in discussions regarding the future of nuclear power and associated nuclear activities. Effective public communication and public participation are often put forward as the key building blocks in garnering public acceptance. A recent international workshop in Finland provided insights into other features that might also be important to building and sustaining public confidence in nuclear activities. The workshop was held in Finland in close cooperation with Finnish stakeholders. This was most appropriate because of the recent successes in achieving positive decisions at the municipal, governmental, and Parliamentary levels, allowing the Finnish high-level radioactive waste repository program to proceed, including the identification and approval of a proposed candidate repository site Much of the workshop discussion appropriately focused on the roles of public participation and public communications in building public confidence. It was clear that well constructed and implemented programs of public involvement and communication and a sense of fairness were essential in building the extent of public confidence needed to allow the repository program in Finland to proceed. It was also clear that there were a number of other elements beyond public involvement that contributed substantially to the success in Finland to date. And, in fact, it appeared that these other factors were also necessary to achieving the Finnish public acceptance. In other words, successful public participation and communication were necessary but not sufficient. What else was important? Culture, politics, and history vary from country to country, providing differing contexts for establishing and maintaining public confidence. What works in one country will not necessarily be effective in another. Nonetheless, there appear to be certain elements that might be common to programs that are successful in sustaining public confidence, and some of

  2. Building Public Confidence in Nuclear Activities

    SciTech Connect

    Isaacs, T

    2002-03-27

    Achieving public acceptance has become a central issue in discussions regarding the future of nuclear power and associated nuclear activities. Effective public communication and public participation are often put forward as the key building blocks in garnering public acceptance. A recent international workshop in Finland provided insights into other features that might also be important to building and sustaining public confidence in nuclear activities. The workshop was held in Finland in close cooperation with Finnish stakeholders. This was most appropriate because of the recent successes in achieving positive decisions at the municipal, governmental, and Parliamentary levels, allowing the Finnish high-level radioactive waste repository program to proceed, including the identification and approval of a proposed candidate repository site. Much of the workshop discussion appropriately focused on the roles of public participation and public communications in building public confidence. It was clear that well constructed and implemented programs of public involvement and communication and a sense of fairness were essential in building the extent of public confidence needed to allow the repository program in Finland to proceed. It was also clear that there were a number of other elements beyond public involvement that contributed substantially to the success in Finland to date. And, in fact, it appeared that these other factors were also necessary to achieving the Finnish public acceptance. In other words, successful public participation and communication were necessary but not sufficient. What else was important? Culture, politics, and history vary from country to country, providing differing contexts for establishing and maintaining public confidence. What works in one country will not necessarily be effective in another. Nonetheless, there appear to be certain elements that might be common to programs that are successful in sustaining public confidence and some of

  3. Nuclear import of the Drosophila Rel protein Dorsal is regulated by phosphorylation.

    PubMed

    Drier, E A; Huang, L H; Steward, R

    1999-03-01

    In Drosophila, dorsal-ventral polarity is determined by a maternally encoded signal transduction pathway that culminates in the graded nuclear localization of the Rel protein, Dorsal. Dorsal is retained in the cytoplasm by the IkappaB protein, Cactus. Signal-dependent phosphorylation of Cactus results in the degradation of Cactus and the nuclear targeting of Dorsal. We present an in-depth study of the functional importance of Dorsal phosphorylation. We find that Dorsal is phosphorylated by the ventral signal while associated with Cactus, and that Dorsal phosphorylation is essential for its nuclear import. In vivo phospholabeling of Dorsal is limited to serine residues in both ovaries and early embryos. A protein bearing mutations in six conserved serines abolishes Dorsal activity, is constitutively cytoplasmic, and appears to eliminate Dorsal phosphorylation, but still interacts with Cactus. Two individual serine-to-alanine mutations produce unexpected results. In a wild-type signaling background, a mutation in the highly conserved PKA site (S312) produces only a weak loss-of-function; however, it completely destabilizes the protein in a cactus mutant background. Significantly, the phosphorylation of another completely conserved serine (S317) regulates the high level of nuclear import found in ventral cells. We conclude that the formation of a wild-type Dorsal nuclear gradient requires the phosphorylation of both Cactus and Dorsal. The strong conservation of the serines suggests that phosphorylation of other Rel proteins is essential for their proper nuclear targeting.

  4. The N-terminal nuclear localization sequences of liver X receptors alpha and beta bind to importin alpha and are essential for both nuclear import and transactivating functions.

    PubMed

    Miller, Anna; Crumbley, Christine; Prüfer, Kirsten

    2009-04-01

    Liver X receptors (LXRs) alpha and beta are nuclear receptors, which form obligate heterodimers with the retinoid X receptor (RXR). The LXRs regulate both redundantly and non-redundantly the transcription of genes controlling cholesterol metabolism and transport as well as lipogenesis. Previously, we showed that mutations in putative N-terminal nuclear localization sequences (NLSs) within both LXRs inhibit nuclear import. Through in vitro studies, we show here that these NLSs bind importin alpha and are both necessary and sufficient for the nuclear import of LXRs. Imaging, transactivation, and electro-mobility shift experiments show that RXR rescues the nuclear import of the LXRalpha NLS mutant yet does not restore its transcriptional activity despite intact DNA binding. In contrast, RXR partially rescues the import of the LXRbeta NLS mutant, but has no effect on its transcriptional activity due to the loss of DNA binding. Experiments with NLS mutant RXR confirmed that RXR may dominate the nuclear import of the RXR/LXRalpha heterodimer, whereas LXRbeta dominates the nuclear import of the RXR/LXRbeta heterodimer. Intriguingly, our data indicate differences between LXRalpha and LXRbeta in their interaction with RXR and in the role their NLSs play in transactivating functions independent of nuclear import.

  5. Nuclear Import of Cdk/Cyclin Complexes: Identification of Distinct Mechanisms for Import of Cdk2/Cyclin E and Cdc2/Cyclin B1

    PubMed Central

    Moore, Jonathan D.; Yang, Jing; Truant, Ray; Kornbluth, Sally

    1999-01-01

    Reversible phosphorylation of nuclear proteins is required for both DNA replication and entry into mitosis. Consequently, most cyclin-dependent kinase (Cdk)/cyclin complexes are localized to the nucleus when active. Although our understanding of nuclear transport processes has been greatly enhanced by the recent identification of nuclear targeting sequences and soluble nuclear import factors with which they interact, the mechanisms used to target Cdk/cyclin complexes to the nucleus remain obscure; this is in part because these proteins lack obvious nuclear localization sequences. To elucidate the molecular mechanisms responsible for Cdk/cyclin transport, we examined nuclear import of fluorescent Cdk2/cyclin E and Cdc2/cyclin B1 complexes in digitonin-permeabilized mammalian cells and also examined potential physical interactions between these Cdks, cyclins, and soluble import factors. We found that the nuclear import machinery recognizes these Cdk/cyclin complexes through direct interactions with the cyclin component. Surprisingly, cyclins E and B1 are imported into nuclei via distinct mechanisms. Cyclin E behaves like a classical basic nuclear localization sequence–containing protein, binding to the α adaptor subunit of the importin-α/β heterodimer. In contrast, cyclin B1 is imported via a direct interaction with a site in the NH2 terminus of importin-β that is distinct from that used to bind importin-α. PMID:9922449

  6. Screening in Low Energy Nuclear Reactions of Importance to Astrophysics

    NASA Astrophysics Data System (ADS)

    Miley, George H.; Hora, Heinz; Luo, Nie

    2004-05-01

    Recent experiments in the LUNAR (Laboratory for Underground Nuclear Astrophysics) project have shown anonymously high electron screening may occur during acceleration driven low energy (<400 kV) ion bombardment of solid targets [1]. These effects become particularly important for E/ Ue < 100 (here E= ion energy and Ue = electron-screening potential energy). Thus these effects become significant for the understanding of reactions involved in nucleosynthesis of the elements and the interpretation of astrophysical data [1]. Another example of the behavior is the surprising threshold behavior near 18 keV for deuterons stopping in 3He gas at energies below the Bragg peak [2]. As pointed out in ref [1], the theoretical explanation for these effects is still under debate. Several researchers have proposed variations of the Trojan Horse Method (THM) to explain these effects [3]. In this paper, we propose an alternate mechanism associated with electron charge accumulation around the target atoms arising from the solid-state structure of the host. This concept will be explained in terms of density functional calculations of charge density profiles in a target undergoing ion dynamic effects [4]. REFERENCES [1] F. Strieder, et al., Naturwissenschaften (2000)88:461-467 [2] A. Formicola, et al., (2000) Eur Phys J. A 8:443-446 [3] S. Typel and H H Wolter, (2000) Few-Body System 29:75-93 [4] G. Miley and H. Hora, (2000) Nuclear Reactions in Solids, APS mtg. Lansing, MI [5] G. Miley, A. Lipson, N. Luo, and H. Hora, (2003) IEEE NSS/MIC Conf., Portland, OR

  7. Multiple Novel Signals Mediate Thyroid Hormone Receptor Nuclear Import and Export*

    PubMed Central

    Mavinakere, Manohara S.; Powers, Jeremy M.; Subramanian, Kelly S.; Roggero, Vincent R.; Allison, Lizabeth A.

    2012-01-01

    Thyroid hormone receptor (TR) is a member of the nuclear receptor superfamily that shuttles between the cytosol and nucleus. The fine balance between nuclear import and export of TR has emerged as a critical control point for modulating thyroid hormone-responsive gene expression; however, sequence motifs of TR that mediate shuttling are not fully defined. Here, we characterized multiple signals that direct TR shuttling. Along with the known nuclear localization signal in the hinge domain, we identified a novel nuclear localization signal in the A/B domain of thyroid hormone receptor α1 that is absent in thyroid hormone receptor β1 and inactive in the oncoprotein v-ErbA. Our prior studies showed that thyroid hormone receptor α1 exits the nucleus through two pathways, one dependent on the export factor CRM1 and the other CRM1-independent. Here, we identified three novel CRM1-independent nuclear export signal (NES) motifs in the ligand-binding domain as follows: a highly conserved NES in helix 12 (NES-H12) and two additional NES sequences spanning helix 3 and helix 6, respectively. Mutations predicted to disrupt the α-helical structure resulted in a significant decrease in NES-H12 activity. The high degree of conservation of helix 12 suggests that this region may function as a key NES in other nuclear receptors. Furthermore, our mutagenesis studies on NES-H12 suggest that altered shuttling of thyroid hormone receptor β1 may be a contributing factor in resistance to thyroid hormone syndrome. Taken together, our findings provide a detailed mechanistic understanding of the multiple signals that work together to regulate TR shuttling and transcriptional activity, and they provide important insights into nuclear receptor function in general. PMID:22815488

  8. Nuclear import of a lipid-modified transcription factor

    PubMed Central

    Eisenhaber, Birgit; Sammer, Michaela; Lua, Wai Heng; Benetka, Wolfgang; Liew, Lai Ling; Yu, Weimiao; Lee, Hwee Kuan; Koranda, Manfred; Adhikari, Sharmila

    2011-01-01

    Lipid-modified transcription factors (TFs) are biomolecular oddities, since their reduced mobility and membrane attachment appear to contradict nuclear import required for their gene-regulatory function. NFAT5 isoform a (selected from an in silico screen for predicted lipid-modified TFs) is shown to contribute about half of all endogenous expression of human NFAT5 isoforms in the isotonic state. Wild-type NFAT5a protein is, indeed, myristoylated and palmitoylated on its transport to the plasmalemma via the endoplasmic reticulum and the Golgi. In contrast, its lipid anchor-deficient mutants as well as isoforms NFAT5b/c are diffusely localized in the cytoplasm without preference to vesicular structures. Quantitative/live microscopy shows the plasma membrane-bound fraction of NFAT5a moving into the nucleus upon osmotic stress despite the lipid anchoring. The mobilization mechanism is not based on proteolytic processing of the lipid-anchored N terminus but appears to involve reversible palmitoylation. Thus, NFAT5a is an example of TFs immobilized with lipid anchors at cyotoplasmic membranes in the resting state and that, nevertheless, can translocate into the nucleus upon signal induction. PMID:22071693

  9. Ab initio calculations of nuclear reactions important for astrophysics

    NASA Astrophysics Data System (ADS)

    Navratil, Petr; Dohet-Eraly, Jeremy; Calci, Angelo; Horiuchi, Wataru; Hupin, Guillaume; Quaglioni, Sofia

    2016-09-01

    In recent years, significant progress has been made in ab initio nuclear structure and reaction calculations based on input from QCD employing Hamiltonians constructed within chiral effective field theory. One of the newly developed approaches is the No-Core Shell Model with Continuum (NCSMC), capable of describing both bound and scattering states in light nuclei simultaneously. We will present NCSMC results for reactions important for astrophysics that are difficult to measure at relevant low energies, such as 3He(α,γ)7Be and 3H(α,γ)7Li and 11C(p,γ)12N radiative capture, as well as the 3H(d,n)4He fusion. We will also address prospects of calculating the 2H(α,γ)6Li capture reaction within the NCSMC formalism. Prepared in part by LLNL under Contract DE-AC52-07NA27344. Supported by the U.S. DOE, OS, NP, under Work Proposal No. SCW1158, and by the NSERC Grant No. SAPIN-2016-00033. TRIUMF receives funding from the NRC Canada.

  10. Deterring Nuclear Proliferation: The Importance of IAEA Safeguards: A TEXTBOOK

    SciTech Connect

    Rosenthal, M.D.; Fishbone, L.G.; Gallini, L.; Krass, A.; Kratzer, M.; Sanborn, J.; Ward, B.; Wulf, N. A.

    2012-03-13

    Nuclear terrorism and nuclear proliferation are among the most pressing challenges to international peace and security that we face today. Iran and Syria remain in non-compliance with the safeguards requirements of the NPT, and the nuclear ambitions of North Korea remain unchecked. Despite these challenges, the NPT remains a cornerstone of the nuclear non-proliferation regime, and the safeguards implemented by the International Atomic Energy Agency (IAEA) under the NPT play a critical role in deterring nuclear proliferation.How do they work? Where did they come from? And what is their future? This book answers these questions. Anyone studying the field of nuclear non-proliferation will benefit from reading this book, and for anyone entering the field, the book will enable them to get a running start. Part I describes the foundations of the international safeguards system: its origins in the 1930s - when new discoveries in physics made it clear immediately that nuclear energy held both peril and promise - through the entry into force in 1970 of the NPT, which codified the role of IAEA safeguards as a means to verify states NPT commitments not to acquire nuclear weapons. Part II describes the NPT safeguards system, which is based on a model safeguards agreement developed specifically for the NPT, The Structure and Content of Agreements between the Agency and States required in connection with the Treaty on the Non-Proliferation of Nuclear Weapons, which has been published by the IAEA as INFCIRC/153. Part III describes events, especially in South Africa, the DPRK, and Iraq in the early 1990s, that triggered a transformation in the way in which safeguards were conceptualized and implemented.

  11. Important technology considerations for space nuclear power systems

    SciTech Connect

    Kuspa, J.P.; Wahlquist, E.J.; Bitz, D.A.

    1988-03-01

    This paper discusses the technology considerations that guide the development of space nuclear power sources (NPS) by the Department of Energy (DOE) to meet a wide variety of applications. The Department and its predecessor agencies have been developing NPS since the 1950s and producing NPS for spacecraft for the National Aeronautics and Space Administration (NASA) and the Department of Defense (DOD) since the early 1960s. No one nuclear power type, isotope or reactor, will suffice over the entire range of mission power required. Nor is one type of power conversion system, be it static or dynamic, the optimum choice of all space nuclear power system applications. There is a need for DOE, in partnership with its users, NASA and DOD, to develop a variety of types of space nuclear power sources -- isotope-static, isotope-dynamic, reactor-static, and reactor-dynamic -- to meet mission requirements well into the next century. 2 figs., 1 tab.

  12. Antigenic variation in African trypanosomes: the importance of chromosomal and nuclear context in VSG expression control.

    PubMed

    Glover, Lucy; Hutchinson, Sebastian; Alsford, Sam; McCulloch, Richard; Field, Mark C; Horn, David

    2013-12-01

    African trypanosomes are lethal human and animal parasites that use antigenic variation for evasion of host adaptive immunity. To facilitate antigenic variation, trypanosomes dedicate approximately one third of their nuclear genome, including many minichromosomes, and possibly all sub-telomeres, to variant surface glycoprotein (VSG) genes and associated sequences. Antigenic variation requires transcription of a single VSG by RNA polymerase I (Pol-I), with silencing of other VSGs, and periodic switching of the expressed gene, typically via DNA recombination with duplicative translocation of a new VSG to the active site. Thus, telomeric location, epigenetic controls and monoallelic transcription by Pol-I at an extranucleolar site are prominent features of VSGs and their expression, with telomeres, chromatin structure and nuclear organization all making vitally important contributions to monoallelic VSG expression control and switching. We discuss VSG transcription, recombination and replication control within this chromosomal and sub-nuclear context.

  13. Large scale RNAi reveals the requirement of nuclear envelope breakdown for nuclear import of human papillomaviruses.

    PubMed

    Aydin, Inci; Weber, Susanne; Snijder, Berend; Samperio Ventayol, Pilar; Kühbacher, Andreas; Becker, Miriam; Day, Patricia M; Schiller, John T; Kann, Michael; Pelkmans, Lucas; Helenius, Ari; Schelhaas, Mario

    2014-05-01

    A two-step, high-throughput RNAi silencing screen was used to identify host cell factors required during human papillomavirus type 16 (HPV16) infection. Analysis of validated hits implicated a cluster of mitotic genes and revealed a previously undetermined mechanism for import of the viral DNA (vDNA) into the nucleus. In interphase cells, viruses were endocytosed, routed to the perinuclear area, and uncoated, but the vDNA failed to be imported into the nucleus. Upon nuclear envelope perforation in interphase cells HPV16 infection occured. During mitosis, the vDNA and L2 associated with host cell chromatin on the metaphase plate. Hence, we propose that HPV16 requires nuclear envelope breakdown during mitosis for access of the vDNA to the nucleoplasm. The results accentuate the value of genes found by RNAi screens for investigation of viral infections. The list of cell functions required during HPV16 infection will, moreover, provide a resource for future virus-host cell interaction studies.

  14. Using Nuclear Receptor Activity to Stratify Hepatocarcinogens

    PubMed Central

    Shah, Imran; Houck, Keith; Judson, Richard S.; Kavlock, Robert J.; Martin, Matthew T.; Reif, David M.; Wambaugh, John; Dix, David J.

    2011-01-01

    Background Nuclear receptors (NR) are a superfamily of ligand-activated transcription factors that control a range of cellular processes. Persistent stimulation of some NR is a non-genotoxic mechanism of rodent liver cancer with unclear relevance to humans. Here we report on a systematic analysis of new in vitro human NR activity data on 309 environmental chemicals in relationship to their liver cancer-related chronic outcomes in rodents. Results The effects of 309 environmental chemicals on human constitutive androstane receptors (CAR/NR1I3), pregnane X receptor (PXR/NR1I2), aryl hydrocarbon receptor (AhR), peroxisome proliferator-activated receptors (PPAR/NR1C), liver X receptors (LXR/NR1H), retinoic X receptors (RXR/NR2B) and steroid receptors (SR/NR3) were determined using in vitro data. Hepatic histopathology, observed in rodents after two years of chronic treatment for 171 of the 309 chemicals, was summarized by a cancer lesion progression grade. Chemicals that caused proliferative liver lesions in both rat and mouse were generally more active for the human receptors, relative to the compounds that only affected one rodent species, and these changes were significant for PPAR (p0.001), PXR (p0.01) and CAR (p0.05). Though most chemicals exhibited receptor promiscuity, multivariate analysis clustered them into relatively few NR activity combinations. The human NR activity pattern of chemicals weakly associated with the severity of rodent liver cancer lesion progression (p0.05). Conclusions The rodent carcinogens had higher in vitro potency for human NR relative to non-carcinogens. Structurally diverse chemicals with similar NR promiscuity patterns weakly associated with the severity of rodent liver cancer progression. While these results do not prove the role of NR activation in human liver cancer, they do have implications for nuclear receptor chemical biology and provide insights into putative toxicity pathways. More importantly, these findings suggest the

  15. Oxidized LDL enhances stretch-induced smooth muscle cell proliferation through alterations in nuclear protein import.

    PubMed

    Chahine, Mirna N; Dibrov, Elena; Blackwood, David P; Pierce, Grant N

    2012-12-01

    Mechanical stress contributes to hypertension and atherosclerosis partly through the stimulation of vascular smooth muscle cell (VSMC) proliferation. Oxidized low density lipoprotein (oxLDL) is another important atherogenic factor that can increase VSMC proliferation. The purpose of this study was to investigate whether oxLDL could further enhance the proliferative action of mechanical stretch on VSMC, and to determine the mechanism responsible for this interaction. Because nuclear protein import is critical in regulating gene expression, transcription, and cell proliferation, its involvement in the mitogenic effects of oxLDL and mechanical stress was studied. OxLDL enhanced the proliferative effects of mechanical stretch on its own in rabbit aortic VSMC, and induced increases in the expression of HSP60 in an additive manner. Adenoviral-mediated overexpression of HSP60 induced increases in cell proliferation compared with uninfected VSMC. Mechanical stretch and oxLDL stimulated the rate of nuclear protein import in VSMC and increased the expression of nucleoporins. These effects were sensitive to inhibition of the MAPK pathway. We conclude that oxLDL and mechanical stretch have a synergistic effect on VSMC proliferation. This synergistic effect is induced through a stimulation of nuclear protein import via HSP60 and an activation of the MAPK pathway.

  16. A proline-tyrosine nuclear localization signal (PY-NLS) is required for the nuclear import of fission yeast PAB2, but not of human PABPN1.

    PubMed

    Mallet, Pierre-Luc; Bachand, François

    2013-03-01

    Nuclear poly(A)-binding proteins (PABPs) are evolutionarily conserved proteins that play key roles in eukaryotic gene expression. In the fission yeast Schizosaccharomyces pombe, the major nuclear PABP, Pab2, functions in the maturation of small nucleolar RNAs as well as in nuclear RNA decay. Despite knowledge about its nuclear functions, nothing is known about how Pab2 is imported into the nucleus. Here, we show that Pab2 contains a proline-tyrosine nuclear localization signal (PY-NLS) that is necessary and sufficient for its nuclear localization and function. Consistent with the role of karyopherin β2 (Kapβ2)-type receptors in the import of PY-NLS cargoes, we show that the fission yeast ortholog of human Kapβ2, Kap104, binds to recombinant Pab2 and is required for Pab2 nuclear localization. The absence of arginine methylation in a basic region N-terminal to the PY-core motif of Pab2 did not affect its nuclear localization. However, in the context of a sub-optimal PY-NLS, we found that Pab2 was more efficiently targeted to the nucleus in the absence of arginine methylation, suggesting that this modification can affect the import kinetics of a PY-NLS cargo. Although a sequence resembling a PY-NLS motif can be found in the human Pab2 ortholog, PABPN1, our results indicate that neither a functional PY-NLS nor Kapβ2 activity are required to promote entry of PABPN1 into the nucleus of human cells. Our findings describe the mechanism by which Pab2 is imported into the nucleus, providing the first example of a PY-NLS import system in fission yeast. In addition, this study suggests the existence of alternative or redundant nuclear import pathways for human PABPN1.

  17. Examination of psychological variables related to nuclear attitudes and nuclear activism

    SciTech Connect

    Roy, P.J.

    1985-01-01

    It was hypothesized that knowledge about nuclear arms developments would not be correlated with nuclear attitudes, that sense of efficacy would be positively correlated with magnitude of nuclear activism, and that death anxiety would be correlated with high level of nuclear knowledge and anti-nuclear attitudes, but not with sense of power. It was also hypothesized that positive correlations would be found between nuclear activism and political activism, knowledge of nuclear facts, and degree of adherence to anti-nuclear attitudes. One hundred and forty three women and 90 men participated in this questionnaire study. Major findings are as follows. In general, the more people knew about nuclear developments, the more anti-nuclear were their attitudes. Also, regardless of nuclear attitudes, a positive correlation was found between knowledge of nuclear facts and nuclear activism. Death anxiety and powerlessness were not correlated. There was a positive correlation between anxiety and both nuclear knowledge and anti-nuclear attitudes. A strong positive correlation was found between nuclear activism and anti-nuclear attitudes, and between political activism and nuclear activism. Internal locus of control did not correlate significantly with high sense of power or with high degree of nuclear activism.

  18. Determining Important Nuclear Fragmentation Processes for Human Space Explorations

    NASA Technical Reports Server (NTRS)

    Lin, Zi-Wei

    2005-01-01

    Space radiation from cosmic ray particles is a major risk for astronauts in long-term human space explorations such as a permanent moon base or a trip to Mars. Radiation shielding is needed to reduce the radiation hazard to astronauts, and models have been developed in order to evaluate the effectiveness of different shielding materials. In this talk we determine the nuclear fragmentation cross sections which will most affect the radiation risk behind typical radiation shielding materials. As a result, these cross sections need more theoretical studies and accurate experimental measurements in order for us to more precisely predict the radiation risk in human space explorations.

  19. Light-regulated nuclear import and degradation of Arabidopsis phytochrome-A N-terminal fragments.

    PubMed

    Wolf, Iris; Kircher, Stefan; Fejes, Erzsébet; Kozma-Bognár, László; Schäfer, Eberhard; Nagy, Ferenc; Adám, Eva

    2011-02-01

    The photoreceptor phytochrome-A (phyA) regulates germination and seedling establishment by mediating very low fluence (VLFR) and far-red high irradiance (FR-HIR) responses in Arabidopsis thaliana. In darkness, phyA homodimers exist in the biologically inactive Pr form and are localized in the cytoplasm. Light induces formation of the biologically active Pfr form and subsequent rapid nuclear import. PhyA Pfr, in contrast to the Pr form, is labile and has a half-life of ∼30 min. We produced transgenic plants in a phyA-201 null background that express the PHYA-yellow fluorescent protein (YFP) or the PHYA686-YFP-dimerization domain (DD) and PHYA686-YFP-DD-nuclear localization signal (NLS) or PHYA686-YFP-DD-nuclear exclusion signal (NES) fusion proteins. The PHYA686-YFP fusion proteins contained the N-terminal domain of phyA (686 amino acid residues), a short DD and the YFP. Here we report that (i) PHYA686-YFP-DD fusion protein is imported into the nucleus in a light-dependent fashion; (ii) neither of the PHYA686 fusion proteins is functional in FR-HIR and nuclear VLFR; and (iii) the phyA-dependent, blue light-induced inhibition of hypocotyl growth is mediated by the PHYA686-YFP-DD-NES but not by the PHYA686-YFP-DD-NLS and PHYA686-YFP-DD fusion proteins. We demonstrate that (i) light induces degradation of all PHYA N-terminal-containing fusion proteins and (ii) these N-terminal domain-containing fusion proteins including the constitutively nuclear PHYA686-YFP-DD-NLS and predominantly cytoplasmic PHYA686-YFP-DD-NES degrade at comparable rates but markedly more slowly than PHYA-YFP, whereas (iii) light-induced degradation of the native phyA is faster compared with PHYA-YFP.

  20. Proprotein convertases play an important role in regulating PKGI endoproteolytic cleavage and nuclear transport

    PubMed Central

    Kato, Shin; Zhang, Ruiguang

    2013-01-01

    Nitric oxide and cGMP modulate vascular smooth muscle cell (SMC) phenotype by regulating cell differentiation and proliferation. Recent studies suggest that cGMP-dependent protein kinase I (PKGI) cleavage and the nuclear translocation of a constitutively active kinase fragment, PKGIγ, are required for nuclear cGMP signaling in SMC. However, the mechanisms that control PKGI proteolysis are unknown. Inspection of the amino acid sequence of a PKGI cleavage site that yields PKGIγ and a protease database revealed a putative minimum consensus sequence for proprotein convertases (PCs). Therefore we investigated the role of PCs in regulating PKGI proteolysis. We observed that overexpression of PCs, furin and PC5, but not PC7, which are all expressed in SMC, increase PKGI cleavage in a dose-dependent manner in human embryonic kidney (HEK) 293 cells. Moreover, furin-induced proteolysis of mutant PKGI, in which alanines were substituted into the putative PC consensus sequence, was decreased in these cells. In addition, overexpression of furin increased PKGI proteolysis in LoVo cells, which is an adenocarcinoma cell line expressing defective furin without PC activity. Also, expression of α1-PDX, an engineered serpin-like PC inhibitor, reduced PC activity and decreased PKGI proteolysis in HEK293 cells. Last, treatment of low-passage rat aortic SMC with membrane-permeable PC inhibitor peptides decreased cGMP-stimulated nuclear PKGIγ translocation. These data indicate for the first time that PCs have a role in regulating PKGI proteolysis and the nuclear localization of its active cleavage product, which are important for cGMP-mediated SMC phenotype. PMID:23686857

  1. A cytoplasmically anchored nuclear protein interferes specifically with the import of nuclear proteins but not U1 snRNA

    PubMed Central

    1993-01-01

    A cytoplasmically anchored mutant SV40 T antigen, FS T antigen, was shown previously to interfere specifically with the nuclear import of a heterologous nuclear protein, adenovirus 5 fiber protein, in cultured monkey cells (Schneider, J., C. Schindewolf, K. van Zee, and E. Fanning. 1988. Cell. 54:117-125; van Zee, K., F. Appel, and E. Fanning. 1991. Mol. Cell. Biol. 11:5137-5146). In this report, we demonstrate that FS T antigen also interferes with the nuclear import of adenovirus E1A and a peptide-albumin conjugate bearing multiple copies of the T antigen nuclear localization signal, but not with the import of U1 snRNA. A kinetic analysis indicates that nuclear import of the albumin- peptide conjugate is inhibited only when high intracellular concentrations of FS T antigen are reached. After microinjection into the cytoplasm of cultured cells, purified FS T antigen protein does not accumulate at the nuclear periphery, but rather is distributed in a punctate pattern throughout the cytoplasm. These data support a model in which cytoplasmic anchoring of FS T antigen enables the mutant protein to sequester and titrate out a cellular factor which is required for nuclear protein but not U1 snRNA import. PMID:8468344

  2. Identification of the nuclear localisation signal of O-GlcNAc transferase and its nuclear import regulation

    PubMed Central

    Seo, Hyeon Gyu; Kim, Han Byeol; Kang, Min Jueng; Ryum, Joo Hwan; Yi, Eugene C.; Cho, Jin Won

    2016-01-01

    Nucleocytoplasmic O-GlcNAc transferase (OGT) attaches a single GlcNAc to hydroxyl groups of serine and threonine residues. Although the cellular localisation of OGT is important to regulate a variety of cellular processes, the molecular mechanisms regulating the nuclear localisation of OGT is unclear. Here, we characterised three amino acids (DFP; residues 451–453) as the nuclear localisation signal of OGT and demonstrated that this motif mediated the nuclear import of non-diffusible β-galactosidase. OGT bound the importin α5 protein, and this association was abolished when the DFP motif of OGT was mutated or deleted. We also revealed that O-GlcNAcylation of Ser389, which resides in the tetratricopeptide repeats, plays an important role in the nuclear localisation of OGT. Our findings may explain how OGT, which possesses a NLS, exists in the nucleus and cytosol simultaneously. PMID:27713473

  3. Nuclear Science Teaching Aids and Activities.

    ERIC Educational Resources Information Center

    Woodburn, John H.

    This publication is a sourcebook for science teachers. It provides guides for basic laboratory work in nuclear energy, suggesting various teacher and student demonstrations. Ideas for science clubs, science fairs, and project research seminars are presented. Problem-solving activities for both science and mathematics classes are included, as well…

  4. Using Nuclear Receptor Activity to Stratify Hepatocarcinogens

    EPA Science Inventory

    Nuclear receptors (NR) are a superfamily of ligand-activated transcription factors that control a range of cellular processes. Persistent stimulation of some NR is a non-genotoxic mechanism of rodent liver cancer with unclear relevance to humans. Here we report on a systematic an...

  5. Antibodies against 70-kD heat shock cognate protein inhibit mediated nuclear import of karyophilic proteins

    PubMed Central

    1992-01-01

    Previously, we found that anti-DDDED antibodies strongly inhibited in vivo nuclear transport of nuclear proteins and that these antibodies recognized a protein of 69 kD (p69) from rat liver nuclear envelopes that showed specific binding activities to the nuclear location sequences (NLSs) of nucleoplasmin and SV-40 large T-antigen. Here we identified this protein as the 70-kD heat shock cognate protein (hsc70) based on its mass, isoelectric point, cellular localization, and partial amino acid sequences. Competition studies indicated that the recombinant hsc70 expressed in Escherichia coli binds to transport competent SV-40 T-antigen NLS more strongly than to the point mutated transport incompetent mutant NLS. To investigate the possible involvement of hsc70 in nuclear transport, we examined the effect of anti-hsc70 rabbit antibodies on the nuclear accumulation of karyophilic proteins. When injected into the cytoplasm of tissue culture cells, anti-hsc70 strongly inhibited the nuclear import of nucleoplasmin, SV- 40 T-antigen NLS bearing BSA and histone H1. In contrast, anti-hsc70 IgG did not prevent the diffusion of lysozyme or 17.4-kD FITC-dextran into the nuclei. After injection of these antibodies, cells continued RNA synthesis and were viable. These results indicate that hsc70 interacts with NLS-containing proteins in the cytoplasm before their nuclear import. PMID:1332978

  6. The continuing important role of radionuclide generator systems for nuclear medicine.

    PubMed

    Knapp, F F; Mirzadeh, S

    1994-10-01

    In this review, the continuing importance and status of development of radionuclide generator systems for nuclear medicine are discussed. Radioisotope costs and availability are two important factors, and both nuclear reactors and accelerator facilities are required for production of the parent radioisotopes. Radionuclide generator research is currently focused on the development of generators which provide radioisotopes for positron emission tomography (PET) applications and daughter radioisotopes for various therapeutic applications which decay primarily by particle emission. Generator research continues to be influenced by developments and requirements of complementary technologies, such as the increasing availability of PET. In addition, the availability of a wide spectrum of tumor-specific antibodies, fragments, and peptides for radioimmunodiagnosis and radioimmunotherapy has stimulated the need for generator-derived radioisotopes. The advantages of treatment of arthritis of the synovial joints with radioactive particles (radiation synovectomy) may be expected to be of increasing importance as the elderly population increases, and many of these agents are prepared using generator-derived radioisotopes such as yttrium-90 and rhenium-188. Therapeutic use of the "in vivo generator" is a new approach, where the less radiotoxic parent radioisotope is used to prepare tissue-specific therapeutic agents. Following in vivo site localization, decay of the parent provides the daughter for therapy at the target site. The principal foundation of most diagnostic agents will continue to require technetium-99m from the molybdenum-99/technetium-99m ("Moly") generator. With the limited availability of nuclear reactors and facilities necessary for production and processing of fission 99mTc and the significant issues and problems associated with radioactive waste processing, however, the possibility of utilizing lower specific activity 99Mo produced from neutron activation of

  7. Sensing actin dynamics: Structural basis for G-actin-sensitive nuclear import of MAL

    SciTech Connect

    Hirano, Hidemi; Matsuura, Yoshiyuki

    2011-10-22

    Highlights: {yields} MAL has a bipartite NLS that binds to Imp{alpha} in an extended conformation. {yields} Mutational analyses verified the functional significance of MAL-Imp{alpha} interactions. {yields} Induced folding and NLS-masking by G-actins inhibit nuclear import of MAL. -- Abstract: The coordination of cytoskeletal actin dynamics with gene expression reprogramming is emerging as a crucial mechanism to control diverse cellular processes, including cell migration, differentiation and neuronal circuit assembly. The actin-binding transcriptional coactivator MAL (also known as MRTF-A/MKL1/BSAC) senses G-actin concentration and transduces Rho GTPase signals to serum response factor (SRF). MAL rapidly shuttles between the cytoplasm and the nucleus in unstimulated cells but Rho-induced depletion of G-actin leads to MAL nuclear accumulation and activation of transcription of SRF:MAL-target genes. Although the molecular and structural basis of actin-regulated nucleocytoplasmic shuttling of MAL is not understood fully, it is proposed that nuclear import of MAL is mediated by importin {alpha}/{beta} heterodimer, and that G-actin competes with importin {alpha}/{beta} for the binding to MAL. Here we present structural, biochemical and cell biological evidence that MAL has a classical bipartite nuclear localization signal (NLS) in the N-terminal 'RPEL' domain containing Arg-Pro-X-X-X-Glu-Leu (RPEL) motifs. The NLS residues of MAL adopt an extended conformation and bind along the surface groove of importin-{alpha}, interacting with the major- and minor-NLS binding sites. We also present a crystal structure of wild-type MAL RPEL domain in complex with five G-actins. Comparison of the importin-{alpha}- and actin-complexes revealed that the binding of G-actins to MAL is associated with folding of NLS residues into a helical conformation that is inappropriate for importin-{alpha} recognition.

  8. Defective nuclear import of Tpr in Progeria reflects the Ran sensitivity of large cargo transport.

    PubMed

    Snow, Chelsi J; Dar, Ashraf; Dutta, Anindya; Kehlenbach, Ralph H; Paschal, Bryce M

    2013-05-13

    The RanGTPase acts as a master regulator of nucleocytoplasmic transport by controlling assembly and disassembly of nuclear transport complexes. RanGTP is required in the nucleus to release nuclear localization signal (NLS)-containing cargo from import receptors, and, under steady-state conditions, Ran is highly concentrated in the nucleus. We previously showed the nuclear/cytoplasmic Ran distribution is disrupted in Hutchinson-Gilford Progeria syndrome (HGPS) fibroblasts that express the Progerin form of lamin A, causing a major defect in nuclear import of the protein, translocated promoter region (Tpr). In this paper, we show that Tpr import was mediated by the most abundant import receptor, KPNA2, which binds the bipartite NLS in Tpr with nanomolar affinity. Analyses including NLS swapping revealed Progerin did not cause global inhibition of nuclear import. Rather, Progerin inhibited Tpr import because transport of large protein cargoes was sensitive to changes in the Ran nuclear/cytoplasmic distribution that occurred in HGPS. We propose that defective import of large protein complexes with important roles in nuclear function may contribute to disease-associated phenotypes in Progeria.

  9. Nuclear-cytoplasmic trafficking of NTF2, the nuclear import receptor for the RanGTPase, is subjected to regulation.

    PubMed

    Chafe, Shawn C; Pierce, Jacqueline B; Mangroo, Dev

    2012-01-01

    NTF2 is a cytosolic protein responsible for nuclear import of Ran, a small Ras-like GTPase involved in a number of critical cellular processes, including cell cycle regulation, chromatin organization during mitosis, reformation of the nuclear envelope following mitosis, and controlling the directionality of nucleocytoplasmic transport. Herein, we provide evidence for the first time that translocation of the mammalian NTF2 from the nucleus to the cytoplasm to collect Ran in the GDP form is subjected to regulation. Treatment of mammalian cells with polysorbitan monolaurate was found to inhibit nuclear export of tRNA and proteins, which are processes dependent on RanGTP in the nucleus, but not nuclear import of proteins. Inhibition of the export processes by polysorbitan monolaurate is specific and reversible, and is caused by accumulation of Ran in the cytoplasm because of a block in translocation of NTF2 to the cytoplasm. Nuclear import of Ran and the nuclear export processes are restored in polysorbitan monolaurate treated cells overproducing NTF2. Moreover, increased phosphorylation of a phospho-tyrosine protein and several phospho-threonine proteins was observed in polysorbitan monolaurate treated cells. Collectively, these findings suggest that nucleocytoplasmic translocation of NTF2 is regulated in mammalian cells, and may involve a tyrosine and/or threonine kinase-dependent signal transduction mechanism(s).

  10. Nuclear import/export of hRPF1/Nedd4 regulates the ubiquitin-dependent degradation of its nuclear substrates.

    PubMed

    Hamilton, M H; Tcherepanova, I; Huibregtse, J M; McDonnell, D P

    2001-07-13

    The ubiquitin-protein ligase (E3), hRPF1/Nedd4, is a component of the ubiquitin-proteasome pathway responsible for substrate recognition and specificity. Although previously characterized as a regulator of the stability of cytoplasmic proteins, hRPF1/Nedd4 has also been suggested to have a role in the nucleus. However, in light of the cytoplasmic localization of hRPF1/Nedd4, it is unclear whether bona fide nuclear substrates of hRPF1/Nedd4 exist, and if so, what mechanism may allow a cytoplasmic ubiquitin ligase to manifest nuclear activity. Our search for nuclear substrates led to the identification of the human proline-rich transcript, brain-expressed (hPRTB) protein, the ubiquitination and degradation of which is regulated by hRPF1/Nedd4. Interestingly, hPRTB colocalizes with the splicing factor SC35 in nuclear speckles. Finally, we demonstrate that hRPF1/Nedd4 is indeed capable of entering the nucleus; however, the presence of a functional Rev-like nuclear export sequence in hRPF1/Nedd4 ensures a predominant cytoplasmic localization. Cumulatively, these findings highlight a nuclear role for the ubiquitin ligase hRPF1/Nedd4 and underscore cytoplasmic/nuclear localization as an important regulatory component of hRPF1/Nedd4-substrate recognition.

  11. The Nup358-RanGAP complex is required for efficient importin alpha/beta-dependent nuclear import.

    PubMed

    Hutten, Saskia; Flotho, Annette; Melchior, Frauke; Kehlenbach, Ralph H

    2008-05-01

    In vertebrate cells, the nucleoporin Nup358/RanBP2 is a major component of the filaments that emanate from the nuclear pore complex into the cytoplasm. Nup358 forms a complex with SUMOylated RanGAP1, the GTPase activating protein for Ran. RanGAP1 plays a pivotal role in the establishment of a RanGTP gradient across the nuclear envelope and, hence, in the majority of nucleocytoplasmic transport pathways. Here, we investigate the roles of the Nup358-RanGAP1 complex and of soluble RanGAP1 in nuclear protein transport, combining in vivo and in vitro approaches. Depletion of Nup358 by RNA interference led to a clear reduction of importin alpha/beta-dependent nuclear import of various reporter proteins. In vitro, transport could be partially restored by the addition of importin beta, RanBP1, and/or RanGAP1 to the transport reaction. In intact Nup358-depleted cells, overexpression of importin beta strongly stimulated nuclear import, demonstrating that the transport receptor is the most rate-limiting factor at reduced Nup358-concentrations. As an alternative approach, we used antibody-inhibition experiments. Antibodies against RanGAP1 inhibited the enzymatic activity of soluble and nuclear pore-associated RanGAP1, as well as nuclear import and export. Although export could be fully restored by soluble RanGAP, import was only partially rescued. Together, these data suggest a dual function of the Nup358-RanGAP1 complex as a coordinator of importin beta recycling and reformation of novel import complexes.

  12. The Orphan Nuclear Receptor TR4 Is a Vitamin A-activated Nuclear Receptor

    SciTech Connect

    Zhou, X. Edward; Suino-Powell, Kelly M.; Xu, Yong; Chan, Cee-Wah; Tanabe, Osamu; Kruse, Schoen W.; Reynolds, Ross; Engel, James Douglas; Xu, H. Eric

    2015-11-30

    Testicular receptors 2 and 4 (TR2/4) constitute a subgroup of orphan nuclear receptors that play important roles in spermatogenesis, lipid and lipoprotein regulation, and the development of the central nervous system. Currently, little is known about the structural features and the ligand regulation of these receptors. Here we report the crystal structure of the ligand-free TR4 ligand binding domain, which reveals an autorepressed conformation. The ligand binding pocket of TR4 is filled by the C-terminal half of helix 10, and the cofactor binding site is occupied by the AF-2 helix, thus preventing ligand-independent activation of the receptor. However, TR4 exhibits constitutive transcriptional activity on multiple promoters, which can be further potentiated by nuclear receptor coactivators. Mutations designed to disrupt cofactor binding, dimerization, or ligand binding substantially reduce the transcriptional activity of this receptor. Importantly, both retinol and retinoic acid are able to promote TR4 to recruit coactivators and to activate a TR4-regulated reporter. These findings demonstrate that TR4 is a ligand-regulated nuclear receptor and suggest that retinoids might have a much wider regulatory role via activation of orphan receptors such as TR4.

  13. Galaxy interactions and strength of nuclear activity

    NASA Technical Reports Server (NTRS)

    Simkin, S. M.

    1990-01-01

    Analysis of data in the literature for differential velocities and projected separations of nearby Seyfert galaxies with possible companions shows a clear difference in projected separations between type 1's and type 2's. This kinematic difference between the two activity classes reinforces other independent evidence that their different nuclear characteristics are related to a non-nuclear physical distinction between the two classes. The differential velocities and projected separations of the galaxy pairs in this sample yield mean galaxy masses, sizes, and mass to light ratios which are consistent with those found by the statistical methods of Karachentsev. Although the galaxy sample discussed here is too small and too poorly defined to provide robust support for these conclusions, the results strongly suggest that nuclear activity in Seyfert galaxies is associated with gravitational perturbations from companion galaxies, and that there are physical distinctions between the host companions of Seyfert 1 and Seyfert 2 nuclei which may depend both on the environment and the structure of the host galaxy itself.

  14. Emerging evidence of the importance of rapid, non-nuclear estrogen receptor signaling in the cardiovascular system.

    PubMed

    Ueda, Kazutaka; Karas, Richard H

    2013-06-01

    Estrogen receptors are classically known as ligand-activated transcription factors that regulate gene transcription in cells in response to hormone binding. In addition to this "genomic" signaling pathway, a "rapid, non-nuclear" signaling pathway mediated by cell membrane-associated estrogen receptors also has been recognized. Although for many years there was little evidence to support any physiological relevance of rapid-signaling, very recently evidence has been accumulating supporting the importance of the rapid, non-nuclear signaling as potentially critical for the protective effects of estrogen in the cardiovascular system. Better understanding of the rapid, non-nuclear signaling potentially provides an opportunity to design "pathway-specific" selective estrogen receptor modulators capable of differentially regulating non-nuclear vs. genomic effects that may prove useful ultimately as specific therapies for cardiovascular diseases.

  15. Determining Yankee Nuclear Power Station neutron activation

    SciTech Connect

    Heider, K.J.; Morrissey, K.J. )

    1993-01-01

    The Yankee nuclear power station located in Rowe, Massachusetts, permanently ceased power operations on February 26, 1992, after 31 yr of operation. Yankee has since initiated decommissioning planning activities. A significant component of these activities is a determination of the extent of radiological contamination of the Yankee site. Included in this effort was determination of the extent of neutron activation of plant components. This paper describes the determination of the neutron activation of the Yankee reactor vessel, associated internals, and surrounding structures. The Yankee reactor vessel is a 600-MW(thermal) stainless steel-lined, carbon steel vessel with stainless steel internal components designed by Westinghouse. The reactor vessel is surrounded and supported by a carbon steel neutron shield tank that was filled with chromated water during plant operation. A 5-ft-thick concrete biological shield wall surrounds the neutron shield tank. A project is under way to remove the reactor vessel internals from the reactor vessel.

  16. Calculation of Monte Carlo importance functions for use in nuclear-well logging calculations

    SciTech Connect

    Soran, P.D.; McKeon, D.C.; Booth, T.E.; Schlumberger Well Services, Houston, TX; Los Alamos National Lab., NM )

    1989-07-01

    Importance sampling is essential to the timely solution of Monte Carlo nuclear-logging computer simulations. Achieving minimum variance (maximum precision) of a response in minimum computation time is one criteria for the choice of an importance function. Various methods for calculating importance functions will be presented, new methods investigated, and comparisons with porosity and density tools will be shown. 5 refs., 1 tab.

  17. LANSCE nuclear science facilities and activities

    SciTech Connect

    Nelson, Ronald O

    2010-01-01

    Nuclear science activities at the Los Alamos Neutron Science Center (LANSCE) encompass measurements spanning the neutron energy range from thermal to 600 MeV. The neutron sources use spallation of the LANSCE 800 MeV pulsed proton beam with the time-of-flight technique to measure properties of neutron-induced reactions as a function of energy over this large energy range. Current experiments are conducted at the Lujan Center moderated neutron source, the unmoderated WNR target, and with a lead-slowing-down spectrometer. Instruments in use include the DANCE array of BaF{sub 2} scintillators for neutron capture studies, the FIGARO array of liquid scintillator neutron detectors, the GEANIE array of high-resolution HPGe x-ray and gamma-ray detectors, and a number of fission chambers, and other detectors. The LANL capabilities for production and handling of radioactive materials coupled with the neutron sources and detectors at LANSCE are enabling new and challenging measurements for a variety of applications including nuclear energy and nuclear astrophysics. An overview of recent research and examples of results is presented.

  18. An active nuclear retention signal in the glucocorticoid receptor functions as a strong inducer of transcriptional activation.

    PubMed

    Carrigan, Amanda; Walther, Rhian F; Salem, Houssein Abdou; Wu, Dongmei; Atlas, Ella; Lefebvre, Yvonne A; Haché, Robert J G

    2007-04-13

    The glucocorticoid receptor (GR) cycles between a naive chaperone-complexed form in the cytoplasm and a transcriptionally active steroid-bound nuclear form. Nuclear import of GR occurs rapidly and is mediated through the importin alpha/beta karyopherin import pathway. By contrast, nuclear export of GR occurs only slowly under most conditions, despite a dependence on active signaling. In this study we have defined a nuclear retention signal (NRS) in the hinge region of GR that actively opposes the nuclear export of GR as well as the nuclear export mediated through an ectopic CRM1-dependent nuclear export signal (NES). The GR NRS overlaps closely with the basic NL1 nuclear localization signal (NLS) but can be distinguished from NL1 by targeted mutagenesis. Substitution of the classical NLS from SV40 T antigen for the GR NL1 results in a receptor in which nuclear export is accelerated. Remarkably, although the SV40-modified GR remains predominantly nuclear in the presence of steroid and is recruited to transcriptional regulatory regions indistinguishably from wild-type GR, the substitution dramatically weakens the ability of GR to activate transcription of a mouse mammary tumor virus reporter gene. These results suggest that active nuclear retention of GR plays an integral role in glucocorticoid signaling.

  19. Optogenetic Control of Nuclear Protein Import in Living Cells Using Light-Inducible Nuclear Localization Signals (LINuS).

    PubMed

    Wehler, Pierre; Niopek, Dominik; Eils, Roland; Di Ventura, Barbara

    2016-06-02

    Many biological processes are regulated by the timely import of specific proteins into the nucleus. The ability to spatiotemporally control the nuclear import of proteins of interest therefore allows study of their role in a given biological process as well as controlling this process in space and time. The light-inducible nuclear localization signal (LINuS) was developed based on a natural plant photoreceptor that reversibly triggers the import of proteins of interest into the nucleus with blue light. Each LINuS is a small, genetically encoded domain that is fused to the protein of interest at the N or C terminus. These protocols describe how to carry out initial microscopy-based screening to assess which LINuS variant works best with a protein of interest. © 2016 by John Wiley & Sons, Inc.

  20. Nucleoporin Nup358 facilitates nuclear import of Methoprene-tolerant (Met) in an importin β- and Hsp83-dependent manner.

    PubMed

    He, Qianyu; Zhang, Yuanxi; Zhang, Xu; Xu, DanDan; Dong, Wentao; Li, Sheng; Wu, Rui

    2017-02-01

    The bHLH-PAS transcription factor, Methoprene-tolerant (Met)(1), functions as a juvenile hormone (JH) receptor and transduces JH signals by directly binding to E-box like motifs in the regulatory regions of JH response genes. Nuclear localization of Met is crucial for its transcriptional activity. Our previous studies have shown that the chaperone protein Hsp83 facilitates JH-induced Met nuclear import in Drosophila melanogaster. However, the exact molecular mechanisms of Met nuclear transport are not fully elucidated. Using DNA affinity chromatography, we have previously detected binding of the nucleoporin Nup358, in the presence of JH, to the JH response region (JHRR) sequences isolated from the Krüppel-homolog 1 (Kr-h1) promoter. Here, we have demonstrated that Nup358 regulates JH-Hsp83-induced Met nuclear localization. RNAi-mediated knockdown of Nup358 expression in Drosophila fat body perturbs Met nuclear transport during the 3 h after initiation of wandering, when the JH titer is high. The accompanying reduced expression of the transport receptor importin β in Nup358 RNAi flies could be one of the reasons accounting for Met mislocalization. Furthermore, a tetratricopeptide repeat (TPR) domain at the N-terminal end of Nup358 interacts with Hsp83 and is indispensable for Met nuclear localization. Overexpression of the TPR domain in Drosophila fat body prevents Met nuclear localization resulting in a decrease in JHRR-driven reporter activity and Kr-h1 expression. These data show that Nup358 facilitates JH-induced Met nuclear transport in a manner dependent on importin β and Hsp83.

  1. Summary of nuclear fuel reprocessing activities around the world

    SciTech Connect

    Mellinger, P.J.; Harmon, K.M.; Lakey, L.T.

    1984-11-01

    This review of international practices for nuclear fuel reprocessing was prepared to provide a nontechnical summary of the current status of nuclear fuel reprocessing activities around the world. The sources of information are widely varied.

  2. Nuclear import of the pre-integration complex (PIC): the Achilles heel of HIV?

    PubMed

    Piller, S C; Caly, L; Jans, D A

    2003-07-01

    Current treatments against the Aquired immune deficiency syndrome (AIDS) are reasonably effective in reducing the amount of human immunodeficiency virus (HIV) present in infected patients, but their side-effects, and the emergence of drug-resistant HIV strains have intensified the renewed search for novel anti-HIV therapies. An essential step in HIV infection is the integration of the viral genome into the host cell chromosomes within the nucleus. Unlike other retroviruses, HIV can transport its genetic material, in the form of the large nucleoprotein pre-integration complex (PIC), into the nucleus through the intact nuclear envelope (NE). This enables HIV to infect non-dividing cells such as macrophages and microglial cells. Detailed knowledge of the signal-dependent pathways by which cellular proteins and RNAs cross the NE has accumulated in the past decade, but although several different components of the PIC have been implicated in its nuclear import, the mechanism of nuclear entry remains unclear. Since specifically inhibiting PIC nuclear import would undoubtedly block HIV infection in non-dividing cells, this critical step of HIV replication is of great interest as a drug target. This review examines the complex and controversial literature regarding three PIC components--the HIV proteins matrix, integrase and Vpr--proposed to facilitate PIC nuclear import, and existing models of HIV PIC nuclear import. It also suggests approaches to move towards a better understanding of PIC nuclear import, through examining the role of individual PIC components in the context of the intact PIC by direct visualisation, in order to develop new anti-HIV therapeutics.

  3. Impaired nuclear import of mammalian Dlx4 proteins as a consequence of rapid sequence divergence

    SciTech Connect

    Coubrough, Melissa L.; Bendall, Andrew J. . E-mail: abendall@uoguelph.ca

    2006-11-15

    Dlx genes encode a developmentally important family of transcription factors with a variety of functions and sites of action during vertebrate embryogenesis. The murine Dlx4 gene is an enigmatic member of the family; little is known about the normal developmental function(s) of Dlx4. Here, we show that Dlx4 is expressed in the murine placenta and in a trophoblast cell line where the protein localizes to both the nucleus and cytoplasm. Despite the presence of several leucine/valine-rich motifs that match known nuclear export sequences, cytoplasmic Dlx4 is not due to CRM-1-mediated nuclear export. Rather, nuclear import of Dlx4 is compromised by specific residues that flank the nuclear localization signal. One of these residues represents a novel conserved feature of the Dlx4 protein in placental mammals, and the second represents novel variation within mouse Dlx4 isoforms. Comparison of orthologous protein sequences reveals a particularly high rate of non-synonymous change in the coding regions of mammalian Dlx4 genes. Since impaired nuclear localization is unlikely to enhance the function of a nuclear transcription factor, these data point to reduced selection pressure as the basis for the rapid divergence of the Dlx4 gene within the mammalian clade.

  4. Structural basis for the regulation of nuclear import of Epstein-Barr virus nuclear antigen 1 (EBNA1) by phosphorylation of the nuclear localization signal.

    PubMed

    Nakada, Ryohei; Hirano, Hidemi; Matsuura, Yoshiyuki

    2017-02-26

    Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) is expressed in every EBV-positive tumor and is essential for the maintenance, replication, and transcription of the EBV genome in the nucleus of host cells. EBNA1 is a serine phosphoprotein, and it has been shown that phosphorylation of S385 in the nuclear localization signal (NLS) of EBNA1 increases the binding affinity to the nuclear import adaptor importin-α1 as well as importin-α5, and stimulates nuclear import of EBNA1. To gain insights into how phosphorylation of the EBNA1 NLS regulates nuclear import, we have determined the crystal structures of two peptide complexes of importin-α1: one with S385-phosphorylated EBNA1 NLS peptide, determined at 2.0 Å resolution, and one with non-phosphorylated EBNA1 NLS peptide, determined at 2.2 Å resolution. The structures show that EBNA1 NLS binds to the major and minor NLS-binding sites of importin-α1, and indicate that the binding affinity of the EBNA1 NLS to the minor NLS-binding site could be enhanced by phosphorylation of S385 through electrostatic interaction between the phosphate group of phospho-S385 and K392 of importin-α1 (corresponding to R395 of importin-α5) on armadillo repeat 8.

  5. Nuclear import strategies of high-risk HPV18 L2 minor capsid protein

    SciTech Connect

    Klucevsek, K.; Daley, J.; Darshan, M.S.; Bordeaux, J.; Moroianu, J. . E-mail: moroianu@bc.edu

    2006-08-15

    We have investigated the nuclear import strategies of high-risk HPV18 L2 minor capsid protein. HPV18 L2 interacts with Kap {alpha}{sub 2} adapter, and Kap {beta}{sub 2} and Kap {beta}{sub 3} nuclear import receptors. Moreover, binding of RanGTP to either Kap {beta}{sub 2} or Kap {beta}{sub 3} inhibits their interaction with L2, suggesting that these Kap {beta}/L2 complexes are import competent. Mapping studies show that HPV18 L2 contains two NLSs: in the N-terminus (nNLS) and in the C-terminus (cNLS), both of which can independently mediate nuclear import. Both nNLS and cNLS form a complex with Kap {alpha}{sub 2}{beta}{sub 1} heterodimer and mediate nuclear import via a classical pathway. The nNLS is also essential for the interaction of HPV18 L2 with Kap {beta}{sub 2} and Kap {beta}{sub 3}. Interestingly, both nNLS and cNLS interact with the viral DNA and this DNA binding occurs without nucleotide sequence specificity. Together, the data suggest that HPV18 L2 can interact via its NLSs with several Kaps and the viral DNA and may enter the nucleus via multiple import pathways mediated by Kap {alpha}{sub 2}{beta}{sub 1} heterodimers, Kap {beta}{sub 2} and Kap {beta}{sub 3}.

  6. Piperlongumine selectively suppresses ABC-DLBCL through inhibition of NF-κB p65 subunit nuclear import

    SciTech Connect

    Niu, Mingshan; Shen, Yangling; Xu, Xiaoyu; Yao, Yao; Fu, Chunling; Yan, Zhiling; Wu, Qingyun; Cao, Jiang; Sang, Wei; Zeng, Lingyu; Li, Zhenyu; Liu, Xuejiao; and others

    2015-07-10

    Constitutive NF-κB activation is required for survival of activated B cell-like subtype of diffuse large B cell lymphoma (ABC-DLBCL). However, current NF-κB targeting strategies lack cancer cell specificity. Here, we identified a novel inhibitor, piperlongumine, features direct binding to NF-κB p65 subunit and suppression of p65 nuclear import. This was accompanied by NF-κB reporter activity suppression and NF-κB target gene downregulation. Moreover, mutation of Cys{sup 38} to Ser in p65 abolished this effect of piperlongumine on inhibition of p65 nuclear import. Furthermore, we show that piperlongumine selectively inhibited proliferation and induced apoptosis of ABC-DLBCL cells. Most notably, it has been reported that piperlongumine did not affect normal cells even at high doses and was nontoxic to animals. Hence, our current study provides new insight into piperlongumine's mechanism of action and novel approach to ABC-DLBCL target therapy. - Highlights: • Current NF-κB targeting strategies lack cancer cell specificity. • Piperlongumine inhibits NF-κB p65 subunit nuclear import via directly binding to p65. • Piperlongumine selectively inhibits proliferation of ABC-DLBCL cells. • This study provides a novel approach to ABC-DLBCL target therapy.

  7. Occupational Employment in Nuclear-Related Activities, 1981.

    ERIC Educational Resources Information Center

    Baker, Joe G.; Olsen, Kathryn

    Employment in nuclear- and nuclear energy-related activities in 1981 was examined and compared to that in previous years. Survey instruments were returned by 784 establishments. Total 1981 nuclear-related employment was estimated to be 249,500--a growth of 22,600 (10%) workers over the 1977 total. Government-owned, contractor-operated (GOCO)…

  8. Nuclear import of the yeast hexokinase 2 protein requires α/β-importin-dependent pathway.

    PubMed

    Peláez, Rafael; Fernández-García, Paula; Herrero, Pilar; Moreno, Fernando

    2012-01-27

    Hexokinase 2 (Hxk2) from Saccharomyces cerevisiae was one of the first metabolic enzymes described as a multifunctional protein. Hxk2 has a double subcellular localization and role, it functions as a glycolytic enzyme in the cytoplasm and as a regulator of gene transcription of several Mig1-regulated genes in the nucleus. However, the mechanism by which Hxk2 enters in the nucleus was unknown until now. Here, we report that the Hxk2 protein is an import substrate of the carriers α-importin (Kap60 in yeast) and β-importin (Kap95 in yeast). We also show that the Hxk2 nuclear import and the binding of Hxk2 with Kap60 are glucose-dependent and involve one lysine-rich nuclear localization sequence (NLS), located between lysine 6 and lysine 12. Moreover, Kap95 facilitates the recognition of the Hxk2 NLS1 motif by Kap60 and both importins are essential for Hxk2 nuclear import. It is also demonstrated that Hxk2 nuclear import and its binding to Kap95 and Kap60 depend on the Gsp1-GTP/GDP protein levels. Thus, our study uncovers Hxk2 as a new cargo for the α/β-importin pathway of S. cerevisiae.

  9. Nuclear fear and children: the impact of parental nuclear activism, responsivity, and fear

    SciTech Connect

    LaGuardia, M.R.

    1986-01-01

    This study examines the extent to which parental nuclear fear, parental activism, and parental responsivity is associated with children's (age 10) nuclear fear. Other associated variables investigated include: nuclear denial, general anxiety and fear, and the personal characteristics of sex, socio-economic status, and academic aptitude. Findings indicate that children attend to nuclear issues when their parents attend to a significant degree. Children's hopelessness about the arms race is increased as parents' worry about nuclear war increases. Children's fear about not surviving a nuclear war increases as parents' worry about survivability decreases. Children who have more general fears also indicated that they have a high level of hopelessness, pervasive worry, and much concern about being able to survive a nuclear war. Children with a high degree of general anxiety did not indicate high degrees of nuclear fears. Children with high academic aptitude were more knowledgeable about nuclear issues and expressed more fears about the nuclear threat. Boys demonstrated more knowledge about nuclear issues than girls, and girls expressed much more frequent fear and worry about the nuclear threat than boys. Parents of lower socio-economic statues (SES) expressed more denial about the nuclear threat and were more pro-military than the higher SES parents.

  10. Restoration of nuclear-import failure caused by triple A syndrome and oxidative stress

    SciTech Connect

    Kiriyama, Takao; Hirano, Makito Asai, Hirohide; Ikeda, Masanori; Furiya, Yoshiko; Ueno, Satoshi

    2008-10-03

    Triple A syndrome is an autosomal recessive neurological disease, mimicking motor neuron disease, and is caused by mutant ALADIN, a nuclear-pore complex component. We recently discovered that the pathogenesis involved impaired nuclear import of DNA repair proteins, including DNA ligase I and the cerebellar ataxia causative protein aprataxin. Such impairment was overcome by fusing classical nuclear localization signal (NLS) and 137-aa downstream sequence of XRCC1, designated stretched NLS (stNLS). We report here that the minimum essential sequence of stNLS (mstNLS) is residues 239-276, downsized by more than 100 aa. mstNLS enabled efficient nuclear import of DNA repair proteins in patient fibroblasts, functioned under oxidative stress, and reduced oxidative-stress-induced cell death, more effectively than stNLS. The stress-tolerability of mstNLS was also exerted in control fibroblasts and neuroblastoma cells. These findings may help develop treatments for currently intractable triple A syndrome and other oxidative-stress-related neurological diseases, and contribute to nuclear compartmentalization study.

  11. Radioactivity inspection of Taiwan for food products imported from Japan after the Fukushima nuclear accident.

    PubMed

    Chiu, Huang-Sheng; Huang, Ping-Ji; Wuu, Jyi-Lan; Wang, Jeng-Jong

    2013-11-01

    The 3-11 Earthquake occurred in Japan last year had greatly damaged the lives and properties and also caused the core meltdown accident in the Fukushima nuclear power plant followed by the leakage of radioactive materials into biosphere. In order to protect against the detriment of radiation from foods which were imported from Japan, the Institute of Nuclear Energy Research (INER) in Taiwan started to conduct radioactivity inspection of food products from Japan after the accident. A total of about 20,000 samples had been tested from March 24 2011 to March 31 2012.

  12. The nuclear question: rethinking species importance in multi-species animal groups.

    PubMed

    Srinivasan, Umesh; Raza, Rashid Hasnain; Quader, Suhel

    2010-09-01

    1. Animals group for various benefits, and may form either simple single-species groups, or more complex multi-species associations. Multi-species groups are thought to provide anti-predator and foraging benefits to participant individuals. 2. Despite detailed studies on multi-species animal groups, the importance of species in group initiation and maintenance is still rated qualitatively as 'nuclear' (maintaining groups) or 'attendant' (species following nuclear species) based on species-specific traits. This overly simplifies and limits understanding of inherently complex associations, and is biologically unrealistic, because species roles in multi-species groups are: (i) likely to be context-specific and not simply a fixed species property, and (ii) much more variable than this dichotomy indicates. 3. We propose a new view of species importance (measured as number of inter-species associations), along a continuum from 'most nuclear' to 'least nuclear'. Using mixed-species bird flocks from a tropical rainforest in India as an example, we derive inter-species association measures from randomizations on bird species abundance data (which takes into account species 'availability') and data on 86 mixed-species flocks from two different flock types. Our results show that the number and average strength of inter-species associations covary positively, and we argue that species with many, strong associations are the most nuclear. 4. From our data, group size and foraging method are ecological and behavioural traits of species that best explain nuclearity in mixed-species bird flocks. Parallels have been observed in multi-species fish shoals, in which group size and foraging method, as well as diet, have been shown to correlate with nuclearity. Further, the context in which multi-species groups occur, in conjunction with species-specific traits, influences the role played by a species in a multi-species group, and this highlights the importance of extrinsic factors in

  13. Nuclear Structure and Galactic γ-Ray Activity.

    PubMed

    Görres, J

    2000-01-01

    The observation of galactic γ lines following the decay of radioactive nuclei provides a direct link between nuclear physics experiments in earth-based laboratories and astrophysical observations with space-based observatories. Two examples are presented to illustrate this interplay: the measurement of the lifetime of (44)Ti to allow an improved determination of the (44)Ti mass of the supernova remnant Cassiopeia A from the observed γ ray activity and the measurements of excited states in (24)Si to determine the reaction rate of (23)Al(p, γ)(24)Si which might be important for a reduced production of (22)Na in novae.

  14. Establishment of Korean Nuclear Ombudsman System Importing Compensation and Insurance Concept for Residents

    SciTech Connect

    Hyun Seok Ko; Yong Min Kim; Young Wook Lee; Dong Hoon Shin; Young Ho Cho; Chang Sun Kang

    2006-07-01

    not established a firm phase as right relief body to be believed by public with lack of independence, authority and specialty. Therefore the establishment of organization that can be the role as special ombudsman organization about sensitive and special matter like nuclear matter is necessary. Definite establishment element of ombudsman system is to introduce of congress type ombudsman element, to be permanent standing system ombudsman, to limit jurisdiction extent and have strong authority, to be able to access easily, to be composed of legal and nuclear specialist. One of important requirements of resident is compensation. So, based on the resident compensation theory about aversion equipment, ombudsman system should be established introducing the insurance theory through risk management as functional background for appropriate compensation. (authors)

  15. Different pathways for the nuclear import of yeast RNA polymerase II.

    PubMed

    Gómez-Navarro, Natalia; Estruch, Francisco

    2015-11-01

    Recent studies suggest that RNA polymerase II (Pol II) has to be fully assembled before being imported into the nucleus, while other reports indicate a distinct mechanism to import large and small subunits. In yeast, Iwr1 binds to the holoenzyme assembled in the cytoplasm and directs its nuclear entry. However, as IWR1 is not an essential gene, Iwr1-independent pathway(s) for the nuclear import of Pol II must exist. In this paper, we investigate the transport into the nucleus of several large and small Pol II subunits in the mutants of genes involved in Pol II biogenesis. We also analyse subcellular localization in the presence of drugs that can potentially affect Pol II nuclear import. Our results show differences in the cellular distribution between large and small subunits when Pol II biogenesis was impaired. Our data suggest that, in addition to the fully assembled holoenzyme, Pol II subunits can be imported to the nucleus, either independently or as partial assemblies, through different pathways, including passive diffusion for the small subunits.

  16. Fhit Nuclear Import Following EGF Stimulation Sustains Proliferation of Breast Cancer Cells.

    PubMed

    Bianchi, Francesca; Sasso, Marianna; Turdo, Federica; Beretta, Giovanni L; Casalini, Patrizia; Ghirelli, Cristina; Sfondrini, Lucia; Ménard, Sylvie; Tagliabue, Elda; Campiglio, Manuela

    2015-11-01

    The tumor-suppressor protein fragile histidine triad (Fhit) exerts its functions in the cytoplasm, although some reports suggest that it may also act in the nucleus. We previously showed that cytosolic Fhit protein levels in cancer cell lines stimulated to proliferate were reduced by proteasomal degradation. Here, we demonstrate that Fhit is physiologically present in the nucleus of breast cancer cell lines and tissues at a low level and that proliferative stimulation increases nuclear levels. Breast cancer cells expressing the FhitY114F mutant, which do not undergo proteasomal degradation, contained mutated Fhit in the nucleus, while cells treated with a proteasome inhibitor accumulated nuclear Fhit during proliferation. Thus, Fhit nuclear shuttling and proteasome degradation phenomena occur independently. When Fhit was coupled to a nuclear localization sequence, the proliferation rate of the transfected cells increased together with levels of proliferation pathway mediators cyclin D1, phospho-MAPK, and phospho-STAT3. Fhit nuclear translocation upon mitogenic stimulation may represent a new regulatory mechanism that allows rapid restoration of Fhit cytoplasmic levels and promotes the proliferation cascade activated by mitogenic stimulation.

  17. Galaxy interactions and the stimulation of nuclear activity

    NASA Technical Reports Server (NTRS)

    Heckman, Timothy M.

    1990-01-01

    The author discusses the idea that interactions between galaxies can lead to enhanced galactic activity. He discusses whether, apart from the observational evidence, there is a strong theoretical or heuristic motivation for investigating galaxy interactions as stimulators of nuclear activity in galaxies. Galactic interactions as mechanisms for triggering nuclear starbursts are covered.

  18. KIF5B and Nup358 Cooperatively Mediate the Nuclear Import of HIV-1 during Infection

    PubMed Central

    Dharan, Adarsh; Talley, Sarah; Tripathi, Abhishek; Mamede, João I.; Majetschak, Matthias; Campbell, Edward M.

    2016-01-01

    Following envelope mediated fusion, the HIV-1 core is released into the cytoplasm of the target cell and undergoes a series of trafficking and replicative steps that result in the nuclear import of the viral genome, which ultimately leads to the integration of the proviral DNA into the host cell genome. Previous studies have found that disruption of microtubules, or depletion of dynein or kinesin motors, perturb the normal uncoating and trafficking of the viral genome. Here, we show that the Kinesin-1 motor, KIF5B, induces a relocalization of the nuclear pore component Nup358 into the cytoplasm during HIV-1 infection. This relocalization of NUP358 is dependent on HIV-1 capsid, and NUP358 directly associates with viral cores following cytoplasmic translocation. This interaction between NUP358 and the HIV-1 core is dependent on multiple capsid binding surfaces, as this association is not observed following infection with capsid mutants in which a conserved hydrophobic binding pocket (N74D) or the cyclophilin A binding loop (P90A) is disrupted. KIF5B knockdown also prevents the nuclear entry and infection by HIV-1, but does not exert a similar effect on the N74D or P90A capsid mutants which do not rely on Nup358 for nuclear import. Finally, we observe that the relocalization of Nup358 in response to CA is dependent on cleavage protein and polyadenylation factor 6 (CPSF6), but independent of cyclophilin A. Collectively, these observations identify a previously unappreciated role for KIF5B in mediating the Nup358 dependent nuclear import of the viral genome during infection. PMID:27327622

  19. Nuclear import of Nkx2-2 is mediated by multiple pathways.

    PubMed

    Lin, Wenbo; Xu, PengPeng; Guo, YingYing; Jia, Qingjie; Tao, Tao

    2017-01-22

    Nkx2-2 homeoprotein is essential for the development of the central nervous system and pancreas. Although the nuclear localization signals of Nkx2-2 have been identified, the responsible transport receptor is still unknown. Here, we demonstrate that imp α1 not only interacts with Nkx2-2 but also transports it into the nucleus in vitro by acting together with imp β1. However, the nuclear import of Nkx2-2 in cells was not inhibited in response to knockdown expression of endogenous imp β1 or over-expression of Bimax2. Furthermore, imp β1 and imp 13, but not imp 4, directly interact with Nkx2-2 and are capable of transporting Nkx2-2 in an in vitro import assay. By GST pull-down assay, we demonstrate that mutation of NLS1 or NLS2 has no effect on interaction with imp α1 or imp 13, but significantly reduced binding to imp β1. Thus, the nuclear import of Nkx2-2 is mediated not only by the classical import pathway but also directly by imp β1 or imp 13.

  20. Nuclear import of dimerized ribosomal protein Rps3 in complex with its chaperone Yar1

    PubMed Central

    Mitterer, Valentin; Gantenbein, Nadine; Birner-Gruenberger, Ruth; Murat, Guillaume; Bergler, Helmut; Kressler, Dieter; Pertschy, Brigitte

    2016-01-01

    After their cytoplasmic synthesis, ribosomal proteins need to be transported into the nucleus, where they assemble with ribosomal RNA into pre-ribosomal particles. Due to their physicochemical properties, they need protection from aggregation on this path. Newly synthesized ribosomal protein Rps3 forms a dimer that is associated with one molecule of its specific chaperone Yar1. Here we report that redundant pathways contribute to the nuclear import of Rps3, with the classical importin α/β pathway (Kap60/Kap95 in yeast) constituting a main import route. The Kap60/Kap95 heterodimer mediates efficient nuclear import of Rps3 by recognition of an N-terminal monopartite nuclear localization signal (NLS). This Rps3-NLS is located directly adjacent to the Yar1-binding site and, upon binding of Kap60 to Rps3, Yar1 is displaced from the ribosomal protein in vitro. While Yar1 does not directly interact with Kap60 in vitro, affinity purifications of Yar1 and Rps3, however, revealed that Kap60 is present in the Rps3/Yar1 complex in vivo. Indeed we could reconstitute such a protein complex containing Rps3 and both Yar1 and Kap60 in vitro. Our data suggest that binding of Yar1 to one N-domain and binding of Kap60 to the second N-domain of dimerized Rps3 orchestrates import and protection of the ribosomal protein. PMID:27819319

  1. 77 FR 27113 - Export and Import of Nuclear Equipment and Material; Export of International Atomic Energy Agency...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-09

    ... / Wednesday, May 9, 2012 / Rules and Regulations#0;#0; ] NUCLEAR REGULATORY COMMISSION 10 CFR Part 110 RIN 3150-AJ04 Export and Import of Nuclear Equipment and Material; Export of International Atomic Energy Agency Safeguards Samples AGENCY: Nuclear Regulatory Commission. ACTION: Final rule. SUMMARY: The...

  2. Importin-β facilitates nuclear import of human GW proteins and balances cytoplasmic gene silencing protein levels

    PubMed Central

    Schraivogel, Daniel; Schindler, Susann G.; Danner, Johannes; Kremmer, Elisabeth; Pfaff, Janina; Hannus, Stefan; Depping, Reinhard; Meister, Gunter

    2015-01-01

    MicroRNAs (miRNAs) guide Argonaute (Ago) proteins to distinct target mRNAs leading to translational repression and mRNA decay. Ago proteins interact with a member of the GW protein family, referred to as TNRC6A-C in mammals, which coordinate downstream gene-silencing processes. The cytoplasmic functions of TNRC6 and Ago proteins are reasonably well established. Both protein families are found in the nucleus as well. Their detailed nuclear functions, however, remain elusive. Furthermore, it is not clear which import routes Ago and TNRC6 proteins take into the nucleus. Using different nuclear transport assays, we find that Ago as well as TNRC6 proteins shuttle between the cytoplasm and the nucleus. While import receptors might function redundantly to transport Ago2, we demonstrate that TNRC6 proteins are imported by the Importin-β pathway. Finally, we show that nuclear localization of both Ago2 and TNRC6 proteins can depend on each other suggesting actively balanced cytoplasmic Ago – TNRC6 levels. PMID:26170235

  3. NPP financial and regulatory risks-Importance of a balanced and comprehensive nuclear law for a newcomer country considering nuclear power programme

    NASA Astrophysics Data System (ADS)

    Manan, J. A. N. Abd; Mostafa, N. A.; Salim, M. F.

    2015-04-01

    understanding of various requirements of public, regulators, investors, financial institutions, international community, operator and other important stakeholders. Ambiguities and uncertainties, especially with regards to certain conditions and requirements should be minimised by emulating good practices of experienced nuclear regulators. The imposition of various financial requirements such as funds for decommissioning, radioactive waste management, financial security, nuclear liabilities and licensing fees are necessary, but at the same time the quantum needs to be clearly defined. Concerns on absolute liability of the operators need to be addressed through a creation of necessary and proper nuclear insurance legislations to mitigate operator S nuclear liability obligations and other financial risks. Another major risk to investors is the possibility of public resistance which will not only can hinder the construction but can also stop operation of the nuclear power plant which will contribute to huge losses to investors and countries. This may require a provision in the legislation that provide proper compensation for these situations and at the same time to allow operators to engage in nuclear promotional activities, such as community benefit and public consultation as voluntary initiatives. Through proper planning, research, consultation and execution, the proposed nuclear law shall be able to promote good regulatory practices for public and investors' confidence and benefit. Early involvement of various stakeholders is essential as a platform for regular communications between regulators and interested parties. Stakeholders' participation in the NPP programme and law developments will also promote transparency of the projects while upholding the independency of the regulators.

  4. NPP financial and regulatory risks-Importance of a balanced and comprehensive nuclear law for a newcomer country considering nuclear power programme

    SciTech Connect

    Manan, J. A. N. Abd Mostafa, N. A.; Salim, M. F.

    2015-04-29

    understanding of various requirements of public, regulators, investors, financial institutions, international community, operator and other important stakeholders. Ambiguities and uncertainties, especially with regards to certain conditions and requirements should be minimised by emulating good practices of experienced nuclear regulators. The imposition of various financial requirements such as funds for decommissioning, radioactive waste management, financial security, nuclear liabilities and licensing fees are necessary, but at the same time the quantum needs to be clearly defined. Concerns on absolute liability of the operators need to be addressed through a creation of necessary and proper nuclear insurance legislations to mitigate operator S nuclear liability obligations and other financial risks. Another major risk to investors is the possibility of public resistance which will not only can hinder the construction but can also stop operation of the nuclear power plant which will contribute to huge losses to investors and countries. This may require a provision in the legislation that provide proper compensation for these situations and at the same time to allow operators to engage in nuclear promotional activities, such as community benefit and public consultation as voluntary initiatives. Through proper planning, research, consultation and execution, the proposed nuclear law shall be able to promote good regulatory practices for public and investors’ confidence and benefit. Early involvement of various stakeholders is essential as a platform for regular communications between regulators and interested parties. Stakeholders’ participation in the NPP programme and law developments will also promote transparency of the projects while upholding the independency of the regulators.

  5. Higher Nucleoporin-Importinβ Affinity at the Nuclear Basket Increases Nucleocytoplasmic Import

    PubMed Central

    Azimi, Mohammad; Mofrad, Mohammad R. K.

    2013-01-01

    Several in vitro studies have shown the presence of an affinity gradient in nuclear pore complex proteins for the import receptor Importinβ, at least partially contributing to nucleocytoplasmic transport, while others have historically argued against the presence of such a gradient. Nonetheless, the existence of an affinity gradient has remained an uncharacterized contributing factor. To shed light on the affinity gradient theory and better characterize how the existence of such an affinity gradient between the nuclear pore and the import receptor may influence the nucleocytoplasmic traffic, we have developed a general-purpose agent based modeling (ABM) framework that features a new method for relating rate constants to molecular binding and unbinding probabilities, and used our ABM approach to quantify the effects of a wide range of forward and reverse nucleoporin-Importinβ affinity gradients. Our results indicate that transport through the nuclear pore complex is maximized with an effective macroscopic affinity gradient of 2000 µM, 200 µM and 10 µM in the cytoplasmic, central channel and nuclear basket respectively. The transport rate at this gradient is approximately 10% higher than the transport rate for a comparable pore lacking any affinity gradient, which has a peak transport rate when all nucleoporins have an affinity of 200 µM for Importinβ. Furthermore, this optimal ratio of affinity gradients is representative of the ratio of affinities reported for the yeast nuclear pore complex – suggesting that the affinity gradient seen in vitro is highly optimized. PMID:24282617

  6. In Vivo Islet Protection by a Nuclear Import Inhibitor in a Mouse Model of Type 1 Diabetes

    PubMed Central

    Moore, Daniel J.; Zienkiewicz, Jozef; Kendall, Peggy L.; Liu, Danya; Liu, Xueyan; Veach, Ruth Ann; Collins, Robert D.; Hawiger, Jacek

    2010-01-01

    Background Insulin-dependent Type 1 diabetes (T1D) is a devastating autoimmune disease that destroys beta cells within the pancreatic islets and afflicts over 10 million people worldwide. These patients face life-long risks for blindness, cardiovascular and renal diseases, and complications of insulin treatment. New therapies that protect islets from autoimmune destruction and allow continuing insulin production are needed. Increasing evidence regarding the pathomechanism of T1D indicates that islets are destroyed by the relentless attack by autoreactive immune cells evolving from an aberrant action of the innate, in addition to adaptive, immune system that produces islet-toxic cytokines, chemokines, and other effectors of islet inflammation. We tested the hypothesis that targeting nuclear import of stress-responsive transcription factors evoked by agonist-stimulated innate and adaptive immunity receptors would protect islets from autoimmune destruction. Principal Findings Here we show that a first-in-class inhibitor of nuclear import, cSN50 peptide, affords in vivo islet protection following a 2-day course of intense treatment in NOD mice, which resulted in a diabetes-free state for one year without apparent toxicity. This nuclear import inhibitor precipitously reduces the accumulation of islet-destructive autoreactive lymphocytes while enhancing activation-induced cell death of T and B lymphocytes derived from autoimmune diabetes-prone, non-obese diabetic (NOD) mice that develop T1D. Moreover, in this widely used model of human T1D we noted attenuation of pro-inflammatory cytokine and chemokine production in immune cells. Conclusions These results indicate that a novel form of immunotherapy that targets nuclear import can arrest inflammation-driven destruction of insulin-producing beta cells at the site of autoimmune attack within pancreatic islets during the progression of T1D. PMID:20949090

  7. Bending-Twisting Motions and Main Interactions in Nucleoplasmin Nuclear Import

    PubMed Central

    Takeda, Agnes Alessandra Sekijima; Braz, Antônio Sérgio Kimus; Lemke, Ney

    2016-01-01

    Alpha solenoid proteins play a key role in regulating the classical nuclear import pathway, recognizing a target protein and transporting it into the nucleus. Importin-α (Impα) is the solenoid responsible for cargo protein recognition, and it has been extensively studied by X-ray crystallography to understand the binding specificity. To comprehend the main motions of Impα and to extend the information about the critical interactions during carrier-cargo recognition, we surveyed different conformational states based on molecular dynamics (MD) and normal mode (NM) analyses. Our model of study was a crystallographic structure of Impα complexed with the classical nuclear localization sequence (cNLS) from nucleoplasmin (Npl), which was submitted to multiple 100 ns of MD simulations. Representative conformations were selected for calculating the 87 lowest frequencies NMs of vibration, and a displacement approach was applied along each NM. Based on geometric criteria, using the radius of curvature and inter-repeat angles as the reference metrics, the main motions of Impα were described. Moreover, we determined the salt bridges, hydrogen bonds and hydrophobic interactions in the Impα-NplNLS interface. Our results show the bending and twisting motions participating in the recognition of nuclear proteins, allowing the accommodation and adjustment of a classical bipartite NLS sequence. The essential contacts for the nuclear import were also described and were mostly in agreement with previous studies, suggesting that the residues in the cNLS linker region establish important contacts with Impα adjusting the cNLS backbone. The MD simulations combined with NM analysis can be applied to the Impα-NLS system to help understand interactions between Impα and cNLSs and the analysis of non-classic NLSs. PMID:27258022

  8. Regulation of nuclear pore complex conformation by IP(3) receptor activation.

    PubMed Central

    Moore-Nichols, David; Arnott, Anne; Dunn, Robert C

    2002-01-01

    In recent years, both the molecular architecture and functional dynamics of nuclear pore complexes (NPCs) have been revealed with increasing detail. These large, supramolecular assemblages of proteins form channels that span the nuclear envelope of cells, acting as crucial regulators of nuclear import and export. From the cytoplasmic face of the nuclear envelope, nuclear pore complexes exhibit an eightfold symmetric ring structure encompassing a central lumen. The lumen often appears occupied by an additional structure alternatively referred to as the central granule, nuclear transport complex, or nuclear plug. Previous studies have suggested that the central granule may play a role in mediating calcium-dependent regulation of diffusion across the nuclear envelope for intermediate sized molecules (10-40 kDa). Using atomic force microscopy to measure the surface topography of chemically fixed Xenopus laevis oocyte nuclear envelopes, we present measurements of the relative position of the central granule within the NPC lumen under a variety of conditions known to modify nuclear Ca(2+) stores. These measurements reveal a large, approximately 9-nm displacement of the central granule toward the cytoplasmic face of the nuclear envelope under calcium depleting conditions. Additionally, activation of nuclear inositol triphosphate (IP(3)) receptors by the specific agonist, adenophostin A, results in a concentration-dependent displacement of central granule position with an EC(50) of ~1.2 nM. The displacement of the central granule within the NPC is observed on both the cytoplasmic and nucleoplasmic faces of the nuclear envelope. The displacement is blocked upon treatment with xestospongin C, a specific inhibitor of IP(3) receptor activation. These results extend previous models of NPC conformational dynamics linking central granule position to depletion of IP(3) sensitive nuclear envelope calcium stores. PMID:12202368

  9. The hypoxic regulator of sterol synthesis Nro1 is a nuclear import adaptor

    PubMed Central

    Yeh, Tzu-Lan; Lee, Chih-Yung S.; Amzel, L. Mario; Espenshade, Peter J.; Bianchet, Mario A.

    2011-01-01

    SUMMARY Fission yeast protein Sre1, the homolog of the mammalian sterol regulatory element binding protein (SREBP), is a hypoxic transcription factor required for sterol homeostasis and low oxygen growth. Nro1 regulates the stability of the N-terminal transcription factor domain of Sre1 (Sre1N) by inhibiting the action of the prolyl 4-hydroxylase-like Ofd1 in an oxygen-dependent manner. The crystal structure of Nro1 determined at 2.2 Å resolution shows an all-α-helical fold that can be divided into two domains: a small N-terminal domain and a larger C-terminal HEAT-repeat domain. Follow-up studies showed that Nro1 defines a new class of nuclear import adaptor that functions both in Ofd1 nuclear localization and in the oxygen-dependent inhibition of Ofd1 to control the hypoxic response. PMID:21481773

  10. The Hypoxic Regulator of Sterol Synthesis Nro1 Is a Nuclear Import Adaptor

    SciTech Connect

    T Yeh; C Lee; L Amzel; P Espenshade; M Bianchet

    2011-12-31

    Fission yeast protein Sre1, the homolog of the mammalian sterol regulatory element-binding protein (SREBP), is a hypoxic transcription factor required for sterol homeostasis and low-oxygen growth. Nro1 regulates the stability of the N-terminal transcription factor domain of Sre1 (Sre1N) by inhibiting the action of the prolyl 4-hydroxylase-like Ofd1 in an oxygen-dependent manner. The crystal structure of Nro1 determined at 2.2 {angstrom} resolution shows an all-{alpha}-helical fold that can be divided into two domains: a small N-terminal domain, and a larger C-terminal HEAT-repeat domain. Follow-up studies showed that Nro1 defines a new class of nuclear import adaptor that functions both in Ofd1 nuclear localization and in the oxygen-dependent inhibition of Ofd1 to control the hypoxic response.

  11. A RanGTP-independent mechanism allows ribosomal protein nuclear import for ribosome assembly

    PubMed Central

    Schütz, Sabina; Fischer, Ute; Altvater, Martin; Nerurkar, Purnima; Peña, Cohue; Gerber, Michaela; Chang, Yiming; Caesar, Stefanie; Schubert, Olga T; Schlenstedt, Gabriel; Panse, Vikram G

    2014-01-01

    Within a single generation time a growing yeast cell imports ∼14 million ribosomal proteins (r-proteins) into the nucleus for ribosome production. After import, it is unclear how these intrinsically unstable and aggregation-prone proteins are targeted to the ribosome assembly site in the nucleolus. Here, we report the discovery of a conserved nuclear carrier Tsr2 that coordinates transfer of the r-protein eS26 to the earliest assembling pre-ribosome, the 90S. In vitro studies revealed that Tsr2 efficiently dissociates importin:eS26 complexes via an atypical RanGTP-independent mechanism that terminates the import process. Subsequently, Tsr2 binds the released eS26, shields it from proteolysis, and ensures its safe delivery to the 90S pre-ribosome. We anticipate similar carriers—termed here escortins—to securely connect the nuclear import machinery with pathways that deposit r-proteins onto developing pre-ribosomal particles. DOI: http://dx.doi.org/10.7554/eLife.03473.001 PMID:25144938

  12. The effect of nanoparticle size and NLS density on nuclear targeting in cancer and normal cells; impaired nuclear import and aberrant nanoparticle intracellular trafficking in glioma.

    PubMed

    Tammam, Salma N; Azzazy, Hassan M E; Lamprecht, Alf

    2017-02-27

    The cell nucleus is an interesting target in many diseases with particular interest in cancer. Previously, nuclear targeted small and large chitosan nanoparticles (S-NPs≈25nm, and L-NPs≈150nm respectively), modified with low, intermediate and high densities of NLS (L-NLS, I-NLS and H-NLS) were developed and assessed in L929 fibroblasts. However, to evade apoptosis and stimulate tumor growth cancer cells are capable of manipulating the nuclear-cytoplasmic transport on many levels, making NPs that are capable of nuclear targeting in normal cells incapable of doing so in cancer. For such reason, here, the nuclear delivery efficiency of S-NPs and L-NPs was assessed as a function of their NLS density in cancer and non-cancer cells. For S-NPs, in all cells tested, NLS was unnecessary for nuclear delivery; unmodified S-NPs showed higher nuclear delivery than NLS-S-NPs due to their ability to gain nuclear entry in a passive manner. For L-NPs, L-NLS-L-NPs showed ≈ 8.5, 33, 1.8 and 7.2 fold higher nuclear deliveries than H-NLS-L-NPs in L929 fibroblasts, primary human fibroblasts, HEK 293 and lung cancer cells, respectively. In glioma however, unmodified L-NPs showed highest nuclear delivery, whereas NLS-L-NPs were retained in the cytoplasm. Experiments conducted in the presence of inhibitors of the classical nuclear import pathway indicated that due to overexpression of importin α, classical nuclear import in glioma is impaired leading to aberrant NP intracellular trafficking and nuclear import.

  13. 75 FR 68809 - Agency Information Collection Activities: Importation Bond Structure

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-09

    ... Information Collection Activities: Importation Bond Structure AGENCY: U.S. Customs and Border Protection...: Importation Bond Structure. This is a proposed extension and revision of an information collection that was...: Importation Bond Structure. OMB Number: 1651-0050. Form Numbers: 301 and 5297. Abstract: Bonds are used...

  14. Super-resolution imaging of nuclear import of adeno-associated virus in live cells

    PubMed Central

    Kelich, Joseph M; Ma, Jiong; Dong, Biao; Wang, Qizhao; Chin, Mario; Magura, Connor M; Xiao, Weidong; Yang, Weidong

    2015-01-01

    Adeno-associated virus (AAV) has been developed as a promising human gene therapy vector. Particularly, recombinant AAV vector (rAAV) achieves its transduction of host cells by crossing at least three physiological barriers including plasma membrane, endosomal membrane, and nuclear envelope (NE). So far, the AAV transduction mechanism has not been explored thoroughly at the single viral particle level. In this study, we employed high-speed super-resolution single-point edge-excitation sub-diffraction (SPEED) microscopy to map the events of single rAAV2 particles infecting live human cells with an unprecedented spatiotemporal resolution of 9–12 nm and 2–20 ms. Data reveal that rAAV2 particles are imported through nuclear pore complexes (NPCs) rather than nuclear membrane budding into the nucleus. Moreover, approximately 17% of the rAAV2 molecules starting from the cytoplasm successfully transverse the NPCs to reach the nucleoplasm, revealing that the NPCs act as a strict selective step for AAV delivery. This study lastly suggests a new pathway to improve AAV vectors for human gene therapy. PMID:26665132

  15. NESmapper: accurate prediction of leucine-rich nuclear export signals using activity-based profiles.

    PubMed

    Kosugi, Shunichi; Yanagawa, Hiroshi; Terauchi, Ryohei; Tabata, Satoshi

    2014-09-01

    The nuclear export of proteins is regulated largely through the exportin/CRM1 pathway, which involves the specific recognition of leucine-rich nuclear export signals (NESs) in the cargo proteins, and modulates nuclear-cytoplasmic protein shuttling by antagonizing the nuclear import activity mediated by importins and the nuclear import signal (NLS). Although the prediction of NESs can help to define proteins that undergo regulated nuclear export, current methods of predicting NESs, including computational tools and consensus-sequence-based searches, have limited accuracy, especially in terms of their specificity. We found that each residue within an NES largely contributes independently and additively to the entire nuclear export activity. We created activity-based profiles of all classes of NESs with a comprehensive mutational analysis in mammalian cells. The profiles highlight a number of specific activity-affecting residues not only at the conserved hydrophobic positions but also in the linker and flanking regions. We then developed a computational tool, NESmapper, to predict NESs by using profiles that had been further optimized by training and combining the amino acid properties of the NES-flanking regions. This tool successfully reduced the considerable number of false positives, and the overall prediction accuracy was higher than that of other methods, including NESsential and Wregex. This profile-based prediction strategy is a reliable way to identify functional protein motifs. NESmapper is available at http://sourceforge.net/projects/nesmapper.

  16. TIME-OF-FLIGHT MASS MEASUREMENTS AND THEIR IMPORTANCE FOR NUCLEAR ASTROPHYSICS

    SciTech Connect

    Matos, M.; Shapira, Dan

    2009-01-01

    Atomic masses play an important role in nuclear astrophysics. The lack of experimental values for nuclides of interest has triggered a rapid development of new mass measurement devices around the world, including Time-of-Flight (TOF) mass measurements offering an access to the most exotic nuclides. Recently, the TOF-B rho technique that includes a position measurement for magnetic rigidity correction has been implemented at the NSCL. An experiment with a similar TOF-B rho technique is approved and planned at the next generation radioactive beam facility (RIBF) at RIKEN.

  17. Time-of-Flight Mass Measurements and Their Importance for Nuclear Astrophysics

    NASA Astrophysics Data System (ADS)

    Matoš, M.; Estrade, A.; Amthor, A. M.; Bazin, D.; Becerril, A.; Elliot, T.; Famiano, M.; Gade, A.; Galaviz, D.; Lorusso, G.; Pereira, J.; Portillo, M.; Rogers, A.; Schatz, H.; Shapira, D.; Smith, E.; Stolz, A.; Wallace, M.

    2009-03-01

    Atomic masses play an important role in nuclear astrophysics. The lack of experimental values for nuclides of interest has triggered a rapid development of new mass measurement devices around the world, including Time-of-Flight (TOF) mass measurements offering an access to the most exotic nuclides. Recently, the TOF-Brho technique that includes a position measurement for magnetic rigidity correction has been implemented at the NSCL. An experiment with a similar TOF-Brho technique is approved and planned at the next generation radioactive beam facility (RIBF) at RIKEN.

  18. Safety Oversight of Decommissioning Activities at DOE Nuclear Sites

    SciTech Connect

    Zull, Lawrence M.; Yeniscavich, William

    2008-01-15

    The Defense Nuclear Facilities Safety Board (Board) is an independent federal agency established by Congress in 1988 to provide nuclear safety oversight of activities at U.S. Department of Energy (DOE) defense nuclear facilities. The activities under the Board's jurisdiction include the design, construction, startup, operation, and decommissioning of defense nuclear facilities at DOE sites. This paper reviews the Board's safety oversight of decommissioning activities at DOE sites, identifies the safety problems observed, and discusses Board initiatives to improve the safety of decommissioning activities at DOE sites. The decommissioning of former defense nuclear facilities has reduced the risk of radioactive material contamination and exposure to the public and site workers. In general, efforts to perform decommissioning work at DOE defense nuclear sites have been successful, and contractors performing decommissioning work have a good safety record. Decommissioning activities have recently been completed at sites identified for closure, including the Rocky Flats Environmental Technology Site, the Fernald Closure Project, and the Miamisburg Closure Project (the Mound site). The Rocky Flats and Fernald sites, which produced plutonium parts and uranium materials for defense needs (respectively), have been turned into wildlife refuges. The Mound site, which performed R and D activities on nuclear materials, has been converted into an industrial and technology park called the Mound Advanced Technology Center. The DOE Office of Legacy Management is responsible for the long term stewardship of these former EM sites. The Board has reviewed many decommissioning activities, and noted that there are valuable lessons learned that can benefit both DOE and the contractor. As part of its ongoing safety oversight responsibilities, the Board and its staff will continue to review the safety of DOE and contractor decommissioning activities at DOE defense nuclear sites.

  19. Human GTPases associate with RNA polymerase II to mediate its nuclear import.

    PubMed

    Carré, Clément; Shiekhattar, Ramin

    2011-10-01

    Small GTPases share a biochemical mechanism and act as binary molecular switches. One important function of small GTPases in the cell is nucleocytoplasmic transport of both proteins and RNA. Here, we show the stable association of human GPN1 and GPN3, small GTPases related to Ran, with RNA polymerase II (RNAPII) isolated from either the cytoplasmic or nuclear fraction. GPN1 and GPN3 directly interact with RNAPII subunit 7 (RPB7)/RPB4 and the C-terminal domain (CTD) of RNAPII. Depletion of GPN1 or GPN3 using small interfering RNAs led to decreased RNAPII levels in the nucleus and an accumulation of this enzyme in the cytoplasm of human cells. Furthermore, isolation of a GPN1/GPN3/RNAPII complex from stable cell lines expressing a dominant negative GPN1 harboring mutations in the GTP-binding pocket demonstrated a role for these proteins in nuclear import of RNAPII. Thus, GPN1/GPN3 define a new family of small GTPases that are specialized for the transport of RNA polymerase II into the nucleus.

  20. Sensor Fusion for Nuclear Proliferation Activity Monitoring

    SciTech Connect

    Adel Ghanem, Ph D

    2007-03-30

    The objective of Phase 1 of this STTR project is to demonstrate a Proof-of-Concept (PoC) of the Geo-Rad system that integrates a location-aware SmartTag (made by ZonTrak) and a radiation detector (developed by LLNL). It also includes the ability to transmit the collected radiation data and location information to the ZonTrak server (ZonService). The collected data is further transmitted to a central server at LLNL (the Fusion Server) to be processed in conjunction with overhead imagery to generate location estimates of nuclear proliferation and radiation sources.

  1. Summary of aerospace and nuclear engineering activities

    NASA Technical Reports Server (NTRS)

    1988-01-01

    The Texas A&M Nuclear and Aerospace engineering departments have worked on five different projects for the NASA/USRA Advanced Design Program during the 1987/88 year. The aerospace department worked on two types of lunar tunnelers that would create habitable space. The first design used a heated cone to melt the lunar regolith, and the second used a conventional drill to bore its way through the crust. Both used a dump truck to get rid of waste heat from the reactor as well as excess regolith from the tunneling operation. The nuclear engineering department worked on three separate projects. The NEPTUNE system is a manned, outer-planetary explorer designed with Jupiter exploration as the baseline mission. The lifetime requirement for both reactor and power-conversion systems was twenty years. The second project undertaken for the power supply was a Mars Sample Return Mission power supply. This was designed to produce 2 kW of electrical power for seven years. The design consisted of a General Purpose Heat Source (GPHS) utilizing a Stirling engine as the power conversion unit. A mass optimization was performed to aid in overall design. The last design was a reactor to provide power for propulsion to Mars and power on the surface. The requirements of 300 kW of electrical power output and a mass of less than 10,000 Rg were set. This allowed the reactor and power conversion unit to fit within the Space Shuttle cargo bay.

  2. The nuclear localization of low risk HPV11 E7 protein mediated by its zinc binding domain is independent of nuclear import receptors

    SciTech Connect

    Piccioli, Zachary; McKee, Courtney H.; Leszczynski, Anna; Onder, Zeynep; Hannah, Erin C.; Mamoor, Shahan; Crosby, Lauren; Moroianu, Junona

    2010-11-10

    We investigated the nuclear import of low risk HPV11 E7 protein using 1) transfection assays in HeLa cells with EGFP fusion plasmids containing 11E7 and its domains and 2) nuclear import assays in digitonin-permeabilized HeLa cells with GST fusion proteins containing 11E7 and its domains. The EGFP-11E7 and EGFP-11cE7{sub 39-98} localized mostly to the nucleus. The GST-11E7 and GST-11cE7{sub 39-98} were imported into the nuclei in the presence of either Ran-GDP or RanG19V-GTP mutant and in the absence of nuclear import receptors. This suggests that 11E7 enters the nucleus via a Ran-dependent pathway, independent of nuclear import receptors, mediated by a nuclear localization signal located in its C-terminal domain (cNLS). This cNLS contains the zinc binding domain consisting of two copies of Cys-X-X-Cys motif. Mutagenesis of Cys residues in these motifs changed the localization of the EGFP-11cE7/-11E7 mutants to cytoplasmic, suggesting that the zinc binding domain is essential for nuclear localization of 11E7.

  3. Nuclear Data Activities in Support of the DOE Nuclear Criticality Safety Program

    NASA Astrophysics Data System (ADS)

    Westfall, R. M.; McKnight, R. D.

    2005-05-01

    The DOE Nuclear Criticality Safety Program (NCSP) provides the technical infrastructure maintenance for those technologies applied in the evaluation and performance of safe fissionable-material operations in the DOE complex. These technologies include an Analytical Methods element for neutron transport as well as the development of sensitivity/uncertainty methods, the performance of Critical Experiments, evaluation and qualification of experiments as Benchmarks, and a comprehensive Nuclear Data program coordinated by the NCSP Nuclear Data Advisory Group (NDAG). The NDAG gathers and evaluates differential and integral nuclear data, identifies deficiencies, and recommends priorities on meeting DOE criticality safety needs to the NCSP Criticality Safety Support Group (CSSG). Then the NDAG identifies the required resources and unique capabilities for meeting these needs, not only for performing measurements but also for data evaluation with nuclear model codes as well as for data processing for criticality safety applications. The NDAG coordinates effort with the leadership of the National Nuclear Data Center, the Cross Section Evaluation Working Group (CSEWG), and the Working Party on International Evaluation Cooperation (WPEC) of the OECD/NEA Nuclear Science Committee. The overall objective is to expedite the issuance of new data and methods to the DOE criticality safety user. This paper describes these activities in detail, with examples based upon special studies being performed in support of criticality safety for a variety of DOE operations.

  4. Nuclear import of RNA polymerase II is coupled with nucleocytoplasmic shuttling of the RNA polymerase II-associated protein 2.

    PubMed

    Forget, Diane; Lacombe, Andrée-Anne; Cloutier, Philippe; Lavallée-Adam, Mathieu; Blanchette, Mathieu; Coulombe, Benoit

    2013-08-01

    The RNA polymerase II (RNAP II)-associated protein (RPAP) 2 has been discovered through its association with various subunits of RNAP II in affinity purification coupled with mass spectrometry experiments. Here, we show that RPAP2 is a mainly cytoplasmic protein that shuttles between the cytoplasm and the nucleus. RPAP2 shuttling is tightly coupled with nuclear import of RNAP II, as RPAP2 silencing provokes abnormal accumulation of RNAP II in the cytoplasmic space. Most notably, RPAP4/GPN1 silencing provokes the retention of RPAP2 in the nucleus. Our results support a model in which RPAP2 enters the nucleus in association with RNAP II and returns to the cytoplasm in association with the GTPase GPN1/RPAP4. Although binding of RNAP II to RPAP2 is mediated by an N-terminal domain (amino acids 1-170) that contains a nuclear retention domain, and binding of RPAP4/GPN1 to RPAP2 occurs through a C-terminal domain (amino acids 156-612) that has a dominant cytoplasmic localization domain. In conjunction with previously published data, our results have important implications, as they indicate that RPAP2 controls gene expression by two distinct mechanisms, one that targets RNAP II activity during transcription and the other that controls availability of RNAP II in the nucleus.

  5. Nuclear Concepts & Technological Issues Institute: Teacher Activity Booklet.

    ERIC Educational Resources Information Center

    Davison, Candace C., Ed.; Lunetta, Lois W., Ed.

    For many summers the Radiation Science and Engineering Center at Pennsylvania State University has been the site of a Nuclear Concepts and Technological Issues Institute for secondary school science teachers. As a culminating activity of the institute teachers develop lesson plans, laboratory experiments, demonstrations, or other activities and…

  6. Nuclear data activities at the n_TOF facility at CERN

    NASA Astrophysics Data System (ADS)

    Gunsing, F.; Aberle, O.; Andrzejewski, J.; Audouin, L.; Bécares, V.; Bacak, M.; Balibrea-Correa, J.; Barbagallo, M.; Barros, S.; Bečvář, F.; Beinrucker, C.; Belloni, F.; Berthoumieux, E.; Billowes, J.; Bosnar, D.; Brugger, M.; Caamaño, M.; Calviño, F.; Calviani, M.; Cano-Ott, D.; Cardella, R.; Casanovas, A.; Castelluccio, D. M.; Cerutti, F.; Chen, Y. H.; Chiaveri, E.; Colonna, N.; Cortés-Giraldo, M. A.; Cortés, G.; Cosentino, L.; Damone, L. A.; Deo, K.; Diakaki, M.; Domingo-Pardo, C.; Dressler, R.; Dupont, E.; Durán, I.; Fernández-Domínguez, B.; Ferrari, A.; Ferreira, P.; Finocchiaro, P.; Frost, R. J. W.; Furman, V.; Ganesan, S.; García, A. R.; Gawlik, A.; Gheorghe, I.; Glodariu, T.; Gonçalves, I. F.; González, E.; Goverdovski, A.; Griesmayer, E.; Guerrero, C.; Göbel, K.; Harada, H.; Heftrich, T.; Heinitz, S.; Hernández-Prieto, A.; Heyse, J.; Jenkins, D. G.; Jericha, E.; Käppeler, F.; Kadi, Y.; Katabuchi, T.; Kavrigin, P.; Ketlerov, V.; Khryachkov, V.; Kimura, A.; Kivel, N.; Kokkoris, M.; Krtička, M.; Leal-Cidoncha, E.; Lederer, C.; Leeb, H.; Lerendegui, J.; Licata, M.; Lo Meo, S.; Lonsdale, S. J.; Losito, R.; Macina, D.; Marganiec, J.; Martínez, T.; Masi, A.; Massimi, C.; Mastinu, P.; Mastromarco, M.; Matteucci, F.; Maugeri, E. A.; Mazzone, A.; Mendoza, E.; Mengoni, A.; Milazzo, P. M.; Mingrone, F.; Mirea, M.; Montesano, S.; Musumarra, A.; Nolte, R.; Oprea, A.; Palomo-Pinto, F. R.; Paradela, C.; Patronis, N.; Pavlik, A.; Perkowski, J.; Porras, I.; Praena, J.; Quesada, J. M.; Rajeev, K.; Rauscher, T.; Reifarth, R.; Riego-Perez, A.; Robles, M.; Rout, P.; Radeck, D.; Rubbia, C.; Ryan, J. A.; Sabaté-Gilarte, M.; Saxena, A.; Schillebeeckx, P.; Schmidt, S.; Schumann, D.; Sedyshev, P.; Smith, A. G.; Stamatopoulos, A.; Suryanarayana, S. V.; Tagliente, G.; Tain, J. L.; Tarifeño-Saldivia, A.; Tarrío, D.; Tassan-Got, L.; Tsinganis, A.; Valenta, S.; Vannini, G.; Variale, V.; Vaz, P.; Ventura, A.; Vlachoudis, V.; Vlastou, R.; Wallner, A.; Warren, S.; Weigand, M.; Weiss, C.; Wolf, C.; Woods, P. J.; Wright, T.; Žugec, P.

    2016-10-01

    Nuclear data in general, and neutron-induced reaction cross sections in particular, are important for a wide variety of research fields. They play a key role in the safety and criticality assessment of nuclear technology, not only for existing power reactors but also for radiation dosimetry, medical applications, the transmutation of nuclear waste, accelerator-driven systems, fuel cycle investigations and future reactor systems as in Generation IV. Applications of nuclear data are also related to research fields as the study of nuclear level densities and stellar nucleosynthesis. Simulations and calculations of nuclear technology applications largely rely on evaluated nuclear data libraries. The evaluations in these libraries are based both on experimental data and theoretical models. Experimental nuclear reaction data are compiled on a worldwide basis by the international network of Nuclear Reaction Data Centres (NRDC) in the EXFOR database. The EXFOR database forms an important link between nuclear data measurements and the evaluated data libraries. CERN's neutron time-of-flight facility n_TOF has produced a considerable amount of experimental data since it has become fully operational with the start of the scientific measurement programme in 2001. While for a long period a single measurement station (EAR1) located at 185 m from the neutron production target was available, the construction of a second beam line at 20 m (EAR2) in 2014 has substantially increased the measurement capabilities of the facility. An outline of the experimental nuclear data activities at CERN's neutron time-of-flight facility n_TOF will be presented.

  7. Ebola virus VP24 targets a unique NLS-binding site on karyopherin5 to selectively compete with nuclear import of phosphorylated STAT1

    PubMed Central

    Xu, Wei; Edwards, Megan R.; Borek, Dominika M.; Feagins, Alicia R.; Mittal, Anuradha; Alinger, Joshua B.; Berry, Kayla N.; Yen, Benjamin; Hamilton, Jennifer; Brett, Tom J.; Pappu, Rohit V.; Leung, Daisy W.; Basler, Christopher F.; Amarasinghe, Gaya K.

    2014-01-01

    SUMMARY During anti-viral defense, interferon (IFN) signaling triggers nuclear transport of tyrosine phosphorylated STAT1 (PY-STAT1), which occurs via a subset of karyopherin alpha (KPNA) nuclear transporters. Many viruses, including Ebola virus, actively antagonize STAT1 signaling to counteract the antiviral effects of IFN. Ebola virus VP24 protein (eVP24) binds KPNA to inhibit PY-STAT1 nuclear transport and render cells refractory to IFNs. We describe the structure of human KPNA5 C-terminus in complex with eVP24. In the complex, eVP24 recognizes a unique non-classical nuclear localization signal (NLS) binding site on KPNA5 that is necessary for efficient PY-STAT1 nuclear transport. eVP24 binds KPNA5 with very high affinity to effectively compete with and inhibit PY-STAT1 nuclear transport. In contrast, eVP24 binding does not affect the transport of classical NLS cargo. Thus, eVP24 counters cell-intrinsic innate immunity by selectively targeting PY-STAT1 nuclear import while leaving the transport of other cargo that maybe required for viral replication unaffected. PMID:25121748

  8. Seasonal Pacing - Match Importance Affects Activity in Professional Soccer.

    PubMed

    Link, Daniel; de Lorenzo, Michael F

    2016-01-01

    This research explores the influence of match importance on player activity in professional soccer. Therefore, we used an observational approach and analyzed 1,211 matches of German Bundesliga and 2nd Bundesliga. The importance measurement employed is based on post season consequences of teams involved in a match. This means, if a match result could potentially influence the final rank, and this rank would lead to different consequences for a team, such as qualification for Champions League opposed to qualification for Europe League, then this match is classified as important; otherwise not. Activity was quantified by TOTAL DISTANCE COVERED, SPRINTS, FAST RUNS, DUELS, FOULS and ATTEMPTS. Running parameters were recorded using a semi-automatic optical tracking system, while technical variables were collected by professional data loggers. Based on our importance classification, low important matches occurred at the beginning of round 29. A two-way ANOVA indicates significantly increased FAST RUNS (+4%, d = 0.3), DUELS (+16%, d = 1.0) and FOULS (+36%, d = 1.2) in important matches compared to low important ones. For FAST RUNS and FOULS, this effect only exists in Bundesliga. A comparison of the two leagues show that TOTAL DISTANCE COVERED (+3%, d = 0.9), SPRINTS (+25%, d = 1.4) and FAST RUNS (+15%, d = 1.4) are higher compared to 2nd Bundesliga, whilst FOULS is less in Bundesliga (-7%, d = 0.3). No difference in player activity was found between matches at the beginning of a season (round 1-6) and at the end of a season (round 29-34). We conclude that match importance influences player activity in German professional soccer. The most reasonable explanation is a conscious or unconscious pacing strategy, motivated by preserving abilities or preventing injury. Since this tendency mainly exists in Bundesliga, this may suggest that more skilled players show a higher awareness for the need of pacing.

  9. Seasonal Pacing - Match Importance Affects Activity in Professional Soccer

    PubMed Central

    Link, Daniel; de Lorenzo, Michael F.

    2016-01-01

    This research explores the influence of match importance on player activity in professional soccer. Therefore, we used an observational approach and analyzed 1,211 matches of German Bundesliga and 2nd Bundesliga. The importance measurement employed is based on post season consequences of teams involved in a match. This means, if a match result could potentially influence the final rank, and this rank would lead to different consequences for a team, such as qualification for Champions League opposed to qualification for Europe League, then this match is classified as important; otherwise not. Activity was quantified by TOTAL DISTANCE COVERED, SPRINTS, FAST RUNS, DUELS, FOULS and ATTEMPTS. Running parameters were recorded using a semi-automatic optical tracking system, while technical variables were collected by professional data loggers. Based on our importance classification, low important matches occurred at the beginning of round 29. A two-way ANOVA indicates significantly increased FAST RUNS (+4%, d = 0.3), DUELS (+16%, d = 1.0) and FOULS (+36%, d = 1.2) in important matches compared to low important ones. For FAST RUNS and FOULS, this effect only exists in Bundesliga. A comparison of the two leagues show that TOTAL DISTANCE COVERED (+3%, d = 0.9), SPRINTS (+25%, d = 1.4) and FAST RUNS (+15%, d = 1.4) are higher compared to 2nd Bundesliga, whilst FOULS is less in Bundesliga (-7%, d = 0.3). No difference in player activity was found between matches at the beginning of a season (round 1–6) and at the end of a season (round 29–34). We conclude that match importance influences player activity in German professional soccer. The most reasonable explanation is a conscious or unconscious pacing strategy, motivated by preserving abilities or preventing injury. Since this tendency mainly exists in Bundesliga, this may suggest that more skilled players show a higher awareness for the need of pacing. PMID:27281051

  10. Activation of Keap1/Nrf2 signaling pathway by nuclear epidermal growth factor receptor in cancer cells

    PubMed Central

    Huo, Longfei; Li, Chia-Wei; Huang, Tzu-Hsuan; Lam, Yung Carmen; Xia, Weiya; Tu, Chun; Chang, Wei-Chao; Hsu, Jennifer L; Lee, Dung-Fang; Nie, Lei; Yamaguchi, Hirohito; Wang, Yan; Lang, Jingyu; Li, Long-Yuan; Chen, Chung-Hsuan; Mishra, Lopa; Hung, Mien-Chie

    2014-01-01

    Nuclear translocation of EGFR has been shown to be important for tumor cell growth, survival, and therapeutic resistance. Previously, we detected the association of EGFR with Keap1 in the nucleus. Keap1 is a Kelch-like ECH-associated protein, which plays an important role in cellular response to chemical and oxidative stress by regulating Nrf2 protein stability and nuclear translocation. In this study, we investigate the role of EGFR in regulating Keap1/Nrf2 cascade in the nucleus and provide evidence to show that nuclear EGFR interacts with and phosphorylates nuclear Keap1 to reduce its nuclear protein level. The reduction of nuclear Keap1 consequently stabilizes nuclear Nrf2 and increases its transcriptional activity in cancer cells, which contributes to tumor cell resistance to chemotherapy. PMID:25628777

  11. Structure and Activities of Nuclear Medicine in Kuwait.

    PubMed

    Elgazzar, Abdelhamid H; Owunwanne, Azuwuike; Alenezi, Saud

    2016-07-01

    The practice of nuclear medicine in Kuwait began in 1965 as a clinic for treating thyroid diseases. The practice developed gradually and until 1981 when the Faculty of Medicine established the Division of Nuclear Medicine in the Department of Radiology, which later became a separate department responsible for establishing and managing the practice in all hospitals of Kuwait. In 1987, a nuclear medicine residency program was begun and it is administered by Kuwait Institute for Medical Specializations originally as a 4-year but currently as a 5-year program. Currently there are 11 departments in the ministry of health hospitals staffed by 49 qualified attending physicians, mostly the diplomats of the Kuwait Institute for Medical Specializations nuclear medicine residency program, 4 academic physicians, 2 radiopharmacists, 2 physicists, and 130 technologists. These departments are equipped with 33 dual-head gamma cameras, 10 SPET/CT, 5 PET/CT, 2 cyclotrons, 1 breast-specific gamma imaging, 1 positron-emitting mammography, 10 thyroid uptake units, 8 technegas machines, 7 PET infusion systems, and 8 treadmills. Activities of nuclear medicine in Kuwait include education and training, clinical service, and research. Education includes nuclear medicine technology program in the Faculty of Allied Health Sciences, the 5-year residency program, medical school teaching distributed among different modules of the integrated curriculum with 14 didactic lecture, and other teaching sessions in nuclear medicine MSc program, which run concurrently with the first part of the residency program. The team of Nuclear Medicine in Kuwait has been active in research and has published more than 300 paper, 11 review articles, 12 book chapters, and 17 books in addition to 36 grants and 2 patents. A PhD program approved by Kuwait University Council would begin in 2016.

  12. Nuclear factor Y regulates ancient budgerigar hepadnavirus core promoter activity.

    PubMed

    Shen, Zhongliang; Liu, Yanfeng; Luo, Mengjun; Wang, Wei; Liu, Jing; Liu, Wei; Pan, Shaokun; Xie, Youhua

    2016-09-16

    Endogenous viral elements (EVE) in animal genomes are the fossil records of ancient viruses and provide invaluable information on the origin and evolution of extant viruses. Extant hepadnaviruses include avihepadnaviruses of birds and orthohepadnaviruses of mammals. The core promoter (Cp) of hepadnaviruses is vital for viral gene expression and replication. We previously identified in the budgerigar genome two EVEs that contain the full-length genome of an ancient budgerigar hepadnavirus (eBHBV1 and eBHBV2). Here, we found eBHBV1 Cp and eBHBV2 Cp were active in several human and chicken cell lines. A region from nt -85 to -11 in eBHBV1 Cp was critical for the promoter activity. Bioinformatic analysis revealed a putative binding site of nuclear factor Y (NF-Y), a ubiquitous transcription factor, at nt -64 to -50 in eBHBV1 Cp. The NF-Y core binding site (ATTGG, nt -58 to -54) was essential for eBHBV1 Cp activity. The same results were obtained with eBHBV2 Cp and duck hepatitis B virus Cp. The subunit A of NF-Y (NF-YA) was recruited via the NF-Y core binding site to eBHBV1 Cp and upregulated the promoter activity. Finally, the NF-Y core binding site is conserved in the Cps of all the extant avihepadnaviruses but not of orthohepadnaviruses. Interestingly, a putative and functionally important NF-Y core binding site is located at nt -21 to -17 in the Cp of human hepatitis B virus. In conclusion, our findings have pinpointed an evolutionary conserved and functionally critical NF-Y binding element in the Cps of avihepadnaviruses.

  13. Nuclear waste: Status of DOE`s nuclear waste site characterization activities

    SciTech Connect

    1987-12-31

    Three potential nuclear waste repository sites have been selected to carry out characterization activities-the detailed geological testing to determine the suitability of each site as a repository. The sites are Hanford in south-central Washington State, Yucca Mountain in southern Nevada, and Deaf Smith in the Texas Panhandle. Two key issues affecting the total program are the estimations of the site characterization completion data and costs and DOE`s relationship with the Nuclear Regulatory Commission which has been limited and its relations with affected states and Indian tribes which continue to be difficult.

  14. Nuclear Import of JAK1 Is Mediated by a Classical NLS and Is Required for Survival of Diffuse Large B-cell Lymphoma.

    PubMed

    Zhu, Fen; Hwang, Byounghoon; Miyamoto, Shigeki; Rui, Lixin

    2017-03-01

    JAKs are non-receptor tyrosine kinases that are generally found in association with cytokine receptors. In the canonical pathway, roles of JAKs have well been established in activating STATs in response to cytokine stimulation to modulate gene transcription. In contrast, a noncanonical role of JAK2 has recently been discovered, in which JAK2 in the nucleus imparts the epigenetic regulation of gene transcription through phosphorylation of tyrosine 41 on the histone protein H3. Recent work further demonstrated that this noncanonical mechanism is conserved with JAK1, which is activated by the autocrine cytokines IL6 and IL10 in activated B-cell-like diffuse large B-cell lymphoma (ABC DLBCL), a cancer type that is particularly difficult to treat and has poor prognosis. However, how JAK1 gains access to the nucleus to enable epigenetic regulation remains undefined. Here, we investigated this question and revealed that JAK1 has a classical nuclear localization signal toward the N-terminal region, which can be recognized by multiple importin α isoforms. Moreover, the nuclear import of JAK1 is independent of its kinase activity but is required for the optimal expansion of ABC DLBCL cells in vitroImplications: This study demonstrates that the nuclear import of JAK1 is essential for the optimal fitness of ABC DLBCL cells, and targeting JAK1 nuclear localization is a potential therapeutic strategy for ABC DLBCL. Mol Cancer Res; 15(3); 348-57. ©2016 AACR.

  15. TDP-43 Inhibits NF-κB Activity by Blocking p65 Nuclear Translocation

    PubMed Central

    Zhu, Jingyan; Cynader, Max S.; Jia, William

    2015-01-01

    TDP-43 (TAR DNA binding protein 43) is a heterogeneous nuclear ribonucleoprotein (hnRNP) that has been found to play an important role in neurodegenerative diseases. TDP-43’s involvement in nuclear factor-kappaB pathways has been reported in both neurons and microglial cells. The NF-κB pathway targets hundreds of genes, many of which are involved in inflammation, immunity and cancer. p50/p65 (p50/RelA) heterodimers, as the major Rel complex in the NF-κB family, are induced by diverse external physiological stimuli and modulate transcriptional activity in almost all cell types. Both p65 and TDP-43 translocation occur through the classic nuclear transportation system. In this study, we report that TDP-43 overexpression prevents TNF-α induced p65 nuclear translocation in a dose dependent manner, and that this further inhibits p65 transactivation activity. The inhibition by TDP-43 does not occur through preventing IκB degradation but probably by competing for the nuclear transporter-importin α3 (KPNA4). This competition is dependent on the presence of the nuclear localization signal (NLS) in TDP-43. Silencing TDP-43 using a specific siRNA also increased p65 nuclear localization upon TNF-α stimulation, suggesting that endogenous TDP-43 may be a default suppressor of the NF-κB pathway. Our results indicate that TDP-43 may play an important role in regulating the levels of NF-κB activity by controlling the nuclear translocation of p65. PMID:26571498

  16. Nuclear Receptor Activity and Liver Cancer Lesion Progression

    EPA Science Inventory

    Nuclear receptors (NRs) are ligand-activated transcription factors that control diverse cellular processes. Chronic stimulation of some NRs is a non-genotoxic mechanism of rodent liver cancer with unclear relevance to humans. We explored this question using human CAR, PXR, PPARα,...

  17. The murine Sry gene encodes a nuclear transcriptional activator

    SciTech Connect

    Dubin, R.A.; Ostrer, H.

    1994-09-01

    The Sry gene functions as a genetic switch in gonadal ridge initiating testis determination. The murine Sry and human SRY open reading frames (ORF) share a conserved 79 amino acid motif, the HMG-box, that binds DNA. Outside this region the two genes share no additional homology. These studies were undertaken to determine whether the Sry/SRY genes encode nuclear transcriptional regulators. As judged by the accumulation of lacZ-SRY hybrid proteins in the nucleus, both the human and murine SRY ORFs contain a nuclear localization signal. The murine Sry HMG-box selectively binds the sequence NACAAT in vitro when presented with a random pool of oligonucleotides and binds AACAAT with the highest affinity. The murine Sry ORF, when expressed in HeLa cells, activates transcription of a reporter gene containing multiple copies of the AACAAT binding site. Activation was observed for a GAL4-responsive gene when the murine Sry ORF was linked to the DNA-binding domain of GAL4. Using this system, the activation function was mapped to a C-terminal glutamine/histidine-rich domain. In addition, LexA-Sry fusion genes activated a LexA-responsive gene in yeast. In contrast, a GAL4-human SRY fusion gene did not cause transcriptional activation. These studies suggest that both the human and mouse SRY ORFs encode nuclear, DNA-binding proteins, and that the mouse Sry ORF can function as a transcriptional activator with separable DNA-binding and activator domains.

  18. The RGD integrin binding site in human L1-CAM is important for nuclear signaling

    SciTech Connect

    Gast, Daniela; Riedle, Svenja; Kiefel, Helena; Mueerkoester, Susanne Sebens; Schaefer, Heiner; Schaefer, Michael K.E.; Altevogt, Peter

    2008-08-01

    L1 cell adhesion molecule (L1-CAM) is a transmembrane cell adhesion molecule initially defined as a promigratory molecule in the developing nervous system. L1 is also overexpressed in a variety of human carcinomas and is associated with bad prognosis. In carcinoma cell lines L1 augments cell motility and metastasis, tumor growth in nude mice and induces expression of L1-dependent genes. It is not known whether L1-signaling requires ligand binding. The RGD motif in the sixth Ig domain of L1 is a binding site for integrins. In the present study we analyzed the role of RGDs in L1-signaling using site-directed mutagenesis combined with antibody blocking studies. We observed that L1-RGE expressing HEK293 cells showed reduced cell-cell binding, cell motility, invasiveness and tumor growth in NOD/SCID mice. The RGE-mutation impaired L1-dependent gene regulation and antibodies to {alpha}v{beta}5 integrin had similar effects. Mutant L1 was unable to translocate to the nucleus. Our findings highlight the importance of the RGD site in L1 for human tumors and suggest that nuclear signaling of L1 is dependent on integrins.

  19. Mapping of nuclear import signal and importin {alpha}3 binding regions of 52K protein of bovine adenovirus-3

    SciTech Connect

    Paterson, Carolyn P.; Ayalew, Lisanework E.; Tikoo, Suresh K.

    2012-10-10

    The L1 region of bovine adenovirus (BAdV)-3 encodes a non-structural protein designated 52K. Anti-52K serum detected a protein of 40 kDa, which localized to the nucleus but not to the nucleolus in BAdV-3-infected or transfected cells. Analysis of mutant 52K proteins suggested that three basic residues ({sup 105}RKR{sup 107}) of the identified domain (amino acids {sup 102}GMPRKRVLT{sup 110}) are essential for nuclear localization of 52K. The nuclear import of a GST-52K fusion protein utilizes the classical importin {alpha}/{beta}-dependent nuclear transport pathway. The 52K protein is preferentially bound to the cellular nuclear import receptor importin {alpha}3. Although deletion of amino acid 102-110 is sufficient to abrogate the nuclear localization of 52K, amino acid 90-133 are required for interaction with importin-{alpha}3 and localizing a cytoplasmic protein to the nucleus. These results suggest that 52K contains a bipartite NLS, which preferentially utilize an importin {alpha}3 nuclear import receptor-mediated pathway to transport 52K to the nucleus.

  20. Vesicle-like biomechanics governs important aspects of nuclear geometry in fission yeast.

    PubMed

    Lim H W, Gerald; Huber, Greg; Torii, Yoshihiro; Hirata, Aiko; Miller, Jonathan; Sazer, Shelley

    2007-09-26

    It has long been known that during the closed mitosis of many unicellular eukaryotes, including the fission yeast (Schizosaccharomyces pombe), the nuclear envelope remains intact while the nucleus undergoes a remarkable sequence of shape transformations driven by elongation of an intranuclear mitotic spindle whose ends are capped by spindle pole bodies embedded in the nuclear envelope. However, the mechanical basis of these normal cell cycle transformations, and abnormal nuclear shapes caused by intranuclear elongation of microtubules lacking spindle pole bodies, remain unknown. Although there are models describing the shapes of lipid vesicles deformed by elongation of microtubule bundles, there are no models describing normal or abnormal shape changes in the nucleus. We describe here a novel biophysical model of interphase nuclear geometry in fission yeast that accounts for critical aspects of the mechanics of the fission yeast nucleus, including the biophysical properties of lipid bilayers, forces exerted on the nuclear envelope by elongating microtubules, and access to a lipid reservoir, essential for the large increase in nuclear surface area during the cell cycle. We present experimental confirmation of the novel and non-trivial geometries predicted by our model, which has no free parameters. We also use the model to provide insight into the mechanical basis of previously described defects in nuclear division, including abnormal nuclear shapes and loss of nuclear envelope integrity. The model predicts that (i) despite differences in structure and composition, fission yeast nuclei and vesicles with fluid lipid bilayers have common mechanical properties; (ii) the S. pombe nucleus is not lined with any structure with shear resistance, comparable to the nuclear lamina of higher eukaryotes. We validate the model and its predictions by analyzing wild type cells in which ned1 gene overexpression causes elongation of an intranuclear microtubule bundle that deforms the

  1. Nuclear pore ion channel activity in live syncytial nuclei.

    PubMed

    Bustamante, Jose Omar

    2002-05-01

    Nuclear pore complexes (NPCs) are important nanochannels for the control of gene activity and expression. Most of our knowledge of NPC function has been derived from isolated nuclei and permeabilized cells in cell lysates/extracts. Since recent patch-clamp work has challenged the dogma that NPCs are freely permeable to small particles, a preparation of isolated living nuclei in their native liquid environment was sought and found: the syncytial nuclei in the water of the coconut Cocos nucifera. These nuclei have all properties of NPC-mediated macromolecular transport (MMT) and express foreign green fluorescent protein (GFP) plasmids. They display chromatin movement, are created by particle aggregation or by division, can grow by throwing filaments to catch material, etc. This study shows, for the first time, that living NPCs engaged in MMT do not transport physiological ions - a phenomenon that explains observations of nucleocytoplasmic ion gradients. Since coconuts are inexpensive (less than US$1/nut per litre), this robust preparation may contribute to our understanding of NPCs and cell nucleus and to the development of biotechnologies for the production of DNA, RNA and proteins.

  2. Applying activity-based costing to the nuclear medicine unit.

    PubMed

    Suthummanon, Sakesun; Omachonu, Vincent K; Akcin, Mehmet

    2005-08-01

    Previous studies have shown the feasibility of using activity-based costing (ABC) in hospital environments. However, many of these studies discuss the general applications of ABC in health-care organizations. This research explores the potential application of ABC to the nuclear medicine unit (NMU) at a teaching hospital. The finding indicates that the current cost averages 236.11 US dollars for all procedures, which is quite different from the costs computed by using ABC. The difference is most significant with positron emission tomography scan, 463 US dollars (an increase of 96%), as well as bone scan and thyroid scan, 114 US dollars (a decrease of 52%). The result of ABC analysis demonstrates that the operational time (machine time and direct labour time) and the cost of drugs have the most influence on cost per procedure. Clearly, to reduce the cost per procedure for the NMU, the reduction in operational time and cost of drugs should be analysed. The result also indicates that ABC can be used to improve resource allocation and management. It can be an important aid in making management decisions, particularly for improving pricing practices by making costing more accurate. It also facilitates the identification of underutilized resources and related costs, leading to cost reduction. The ABC system will also help hospitals control costs, improve the quality and efficiency of the care they provide, and manage their resources better.

  3. Practical applications of activation analysis and other nuclear techniques

    SciTech Connect

    Lyon, W S

    1982-01-01

    Neeutron activation analysis (NAA) is a versatile, sensitive multielement, usually nondestructive analytical technique used to determine elemental concentrations in a variety of materials. Samples are irradiated with neutrons in a nuclear reactor, removed, and for the nondestructive technique, the induced radioactivity measured. This measurement of ..gamma.. rays emitted from specific radionuclides makes possible the quantitative determination of elements present. The method is described, advantages and disadvantages listed and a number of examples of its use given. Two other nuclear methods, particle induced x-ray emission and synchrotron produced x-ray fluorescence are also briefly discussed.

  4. Inhibition of nuclear import and cell-cycle progression by mutated forms of the dynamin-like GTPase MxB.

    PubMed

    King, Megan C; Raposo, Graça; Lemmon, Mark A

    2004-06-15

    Mx proteins form a subfamily of the dynamin-like GTPases, which have well established roles in cellular trafficking. Some Mx proteins (e.g., human MxA) have antiviral activity and are tightly regulated by type I IFNs. Others (e.g., human MxB) lack antiviral activity and are thought to have normal cellular functions that remain undefined. Consistent with this hypothesis, we report that MxB is expressed without IFN treatment. MxB seems to be exclusively extranuclear and is concentrated at the cytoplasmic face of nuclear pores, suggesting a role in their regulation. We find that expression of dominant negative (GTPase-defective) MxB mutants efficiently blocks nuclear import and causes a delay in G(1)/S cell-cycle progression. MxB depletion using RNA interference (RNAi) leads to a similar cell-cycle defect but does not block nuclear import. MxB therefore seems not to be required for nuclear import per se but may instead regulate its efficiency and/or kinetics. These studies indicate an unexpected role for a dynamin-like protein in nucleocytoplasmic trafficking and suggest that a related function might be usurped by its antiviral relatives.

  5. Nuclear import of influenza B virus nucleoprotein: Involvement of an N-terminal nuclear localization signal and a cleavage-protection motif

    SciTech Connect

    Wanitchang, Asawin; Narkpuk, Jaraspim; Jongkaewwattana, Anan

    2013-08-15

    The nucleoprotein of influenza B virus (BNP) shares several characteristics with its influenza A virus counterpart (ANP), including localization in the host's nucleus. However, while the nuclear localization signal(s) (NLS) of ANP are well characterized, little is known about those of BNP. In this study, we showed that the fusion protein bearing the BNP N-terminus fused with GFP (N70–GFP) is exclusively nuclear, and identified a highly conserved KRXR motif spanning residues 44–47 as a putative NLS. In addition, we demonstrated that residues 3–15 of BNP, though not an NLS, are also crucial for nuclear import. Results from mutational analyses of N70–GFP and the full-length BNP suggest that this region may be required for protection of the N-terminus from proteolytic cleavage. Altogether, we propose that the N-terminal region of BNP contains the NLS and cleavage-protection motif, which together drive its nuclear localization. - Highlights: • The N-terminal region of BNP is required for nuclear accumulation. • The conserved motif at position 44–47 is a putative nuclear localization signal. • The first 15 amino acids of BNP may function as a cleavage-protection motif. • BNP may get access to the nucleus via a mechanism distinct from ANP.

  6. Alanine Expansions Associated with Congenital Central Hypoventilation Syndrome Impair PHOX2B Homeodomain-mediated Dimerization and Nuclear Import*

    PubMed Central

    Di Lascio, Simona; Belperio, Debora

    2016-01-01

    Heterozygous mutations of the human PHOX2B gene, a key regulator of autonomic nervous system development, lead to congenital central hypoventilation syndrome (CCHS), a neurodevelopmental disorder characterized by a failure in the autonomic control of breathing. Polyalanine expansions in the 20-residues region of the C terminus of PHOX2B are the major mutations responsible for CCHS. Elongation of the alanine stretch in PHOX2B leads to a protein with altered DNA binding, transcriptional activity, and nuclear localization and the possible formation of cytoplasmic aggregates; furthermore, the findings of various studies support the idea that CCHS is not due to a pure loss of function mechanism but also involves a dominant negative effect and/or toxic gain of function for PHOX2B mutations. Because PHOX2B forms homodimers and heterodimers with its paralogue PHOX2A in vitro, we tested the hypothesis that the dominant negative effects of the mutated proteins are due to non-functional interactions with the wild-type protein or PHOX2A using a co-immunoprecipitation assay and the mammalian two-hybrid system. Our findings show that PHOX2B forms homodimers and heterodimerizes weakly with mutated proteins, exclude the direct involvement of the polyalanine tract in dimer formation, and indicate that mutated proteins retain partial ability to form heterodimers with PHOX2A. Moreover, in this study, we investigated the effects of the longest polyalanine expansions on the homeodomain-mediated nuclear import, and our data clearly show that the expanded C terminus interferes with this process. These results provide novel insights into the effects of the alanine tract expansion on PHOX2B folding and activity. PMID:27129232

  7. Induced starburst and nuclear activity: Faith, facts, and theory

    NASA Technical Reports Server (NTRS)

    Shlosman, Isaac

    1990-01-01

    The problem of the origin of starburst and nuclear (nonstellar) activity in galaxies is reviewed. A physical understanding of the mechanism(s) that induce both types of activity requires one to address the following issues: (1) what is the source of fuel that powers starbursts and active galactic nuclei; and (2) how is it channeled towards the central regions of host galaxies? As a possible clue, the author examines the role of non-axisymmetric perturbations of galactic disks and analyzes their potential triggers. Global gravitational instabilities in the gas on scales approx. 100 pc appear to be crucial for fueling the active galactic nuclei.

  8. Graphene oxide as a chemosensitizer: diverted autophagic flux, enhanced nuclear import, elevated necrosis and improved antitumor effects.

    PubMed

    Chen, Guan-Yu; Meng, Chia-Le; Lin, Kuan-Chen; Tuan, Hsing-Yu; Yang, Hong-Jie; Chen, Chiu-Ling; Li, Kuei-Chang; Chiang, Chi-Shiun; Hu, Yu-Chen

    2015-02-01

    Graphene oxide (GO) is a nanomaterial that provokes autophagy in CT26 colon cancer cells and confers antitumor effects. Here we demonstrated that both GO and the chemotherapy drug cisplatin (CDDP) induced autophagy but elicited low degrees of CT26 cell death. Strikingly, GO combined with CDDP (GO/CDDP) potentiated the CT26 cell killing via necrosis. GO/CDDP not only elicited autophagy, but induced the nuclear import of CDDP and the autophagy marker LC3. The nuclear LC3 did not co-localize with p62 or Lamp-2, neither did blocking autolysosome formation significantly hinder the nuclear import of LC3/CDDP and necrosis, indicating that autophagosome and autolysosome formation was dispensable. Conversely, suppressing phagophore formation and importin-α/β significantly alleviated the nuclear import of LC3/CDDP and necrosis. These data suggested that GO/CDDP diverted the LC3 flux in the early phase of autophagy, resulting in LC3 trafficking towards the nucleus in an importin-α/β-dependent manner, which concurred with the CDDP nuclear import and necrosis. Intratumoral injection of GO/CDDP into mice bearing CT26 colon tumors potentiated immune cell infiltration and promoted cell death, autophagy and HMGB1 release, thereby synergistically augmenting the antitumor effects. Altogether, we unveiled a mechanism concerning how nanomaterials chemosensitize cancer cells and demonstrated the potentials of GO as a chemosensitizer.

  9. The conserved amino-terminal domain of hSRP1 alpha is essential for nuclear protein import.

    PubMed Central

    Weis, K; Ryder, U; Lamond, A I

    1996-01-01

    Nuclear proteins are targeted through the nuclear pore complex (NPC) in an energy-dependent reaction. The import reaction is mediated by nuclear localization sequences (NLS) in the substrate which are recognized by heterodimeric cytoplasmic receptors. hSRP1 alpha is an NLS-binding subunit of the human NLS receptor complex and is complexed in vivo with a second subunit of 97 kDa (p97). We show here that a short amino-terminal domain in hSRP1 alpha is necessary and sufficient for its interaction with p97. This domain is conserved in other SRP1-like proteins and its fusion to a cytoplasmic reporter protein is sufficient to promote complete nuclear import, circumventing the usual requirement for an NLS receptor interaction. The same amino-terminal domain inhibits import of NLS-containing proteins when added to an in vitro nuclear transport assay. While full-length hSRP alpha is able to leave the nucleus, the amino-terminal domain alone is not sufficient to promote exit. We conclude that hSRP1 alpha functions as an adaptor to tether NLS-containing substrates to the protein import machinery. Images PMID:8617227

  10. Activation of equine nuclear transfer oocytes: methods and timing of treatment in relation to nuclear remodeling.

    PubMed

    Choi, Young-Ho; Love, Linda B; Westhusin, Mark E; Hinrichs, Katrin

    2004-01-01

    Early development of embryos produced by transfer of equine nuclei to bovine cytoplasts is superior to that of intraspecies equine nuclear transfer embryos. This may be related to differences in chromatin remodeling or efficiency of activation between the two oocyte types. The pattern of donor nucleus remodeling was examined in equine-equine and equine-bovine reconstructed oocytes. Chromosome condensation occurred in equine cytoplasts by 2 h but was not seen in bovine cytoplasts until 4 h. We investigated the effect of activation of equine-equine reconstructed oocytes at <30 min or at 2 h after reconstruction. Four activation treatments were evaluated at each time point: injection of sperm extract alone, or in combination with 6-dimethylaminopurine (6-DMAP), cytochalasin B, or 1% dimethylsulphoxide. There was no significant difference in normal cleavage rate or average nucleus number of embryos between equine oocytes activated <30 min or at 2 h after reconstruction. The combination of 6-DMAP with sperm extract significantly (P < 0.01) improved cleavage rate compared with the other three treatments. Activation with sperm extract and 6-DMAP 2 h after donor nucleus injection gave the highest cleavage (79%) and the highest cleavage with normal nuclei (40%). Sperm extract and 6-DMAP also effectively activated oocytes parthenogenetically, yielding 83% cleavage and 73% cleavage with normal nuclei. These results indicate that although nuclear remodeling occurs rapidly in equine cytoplasts, early activation does not improve embryonic development after reconstruction.

  11. Techniques to evaluate the importance of common cause degradation on reliability and safety of nuclear weapons.

    SciTech Connect

    Darby, John L.

    2011-05-01

    As the nuclear weapon stockpile ages, there is increased concern about common degradation ultimately leading to common cause failure of multiple weapons that could significantly impact reliability or safety. Current acceptable limits for the reliability and safety of a weapon are based on upper limits on the probability of failure of an individual item, assuming that failures among items are independent. We expanded the current acceptable limits to apply to situations with common cause failure. Then, we developed a simple screening process to quickly assess the importance of observed common degradation for both reliability and safety to determine if further action is necessary. The screening process conservatively assumes that common degradation is common cause failure. For a population with between 100 and 5000 items we applied the screening process and conclude the following. In general, for a reliability requirement specified in the Military Characteristics (MCs) for a specific weapon system, common degradation is of concern if more than 100(1-x)% of the weapons are susceptible to common degradation, where x is the required reliability expressed as a fraction. Common degradation is of concern for the safety of a weapon subsystem if more than 0.1% of the population is susceptible to common degradation. Common degradation is of concern for the safety of a weapon component or overall weapon system if two or more components/weapons in the population are susceptible to degradation. Finally, we developed a technique for detailed evaluation of common degradation leading to common cause failure for situations that are determined to be of concern using the screening process. The detailed evaluation requires that best estimates of common cause and independent failure probabilities be produced. Using these techniques, observed common degradation can be evaluated for effects on reliability and safety.

  12. Is knowledge important? Empirical research on nuclear risk communication in two countries.

    PubMed

    Perko, Tanja; Zeleznik, Nadja; Turcanu, Catrinel; Thijssen, Peter

    2012-06-01

    Increasing audience knowledge is often set as a primary objective of risk communication efforts. But is it worthwhile focusing risk communication strategies solely on enhancing specific knowledge? The main research questions tackled in this paper were: (1) if prior audience knowledge related to specific radiation risks is influential for the perception of these risks and the acceptance of communicated messages and (2) if gender, attitudes, risk perception of other radiation risks, confidence in authorities, and living in the vicinity of nuclear/radiological installations may also play an important role in this matter. The goal of this study was to test empirically the mentioned predictors in two independent case studies in different countries. The first case study was an information campaign for iodine pre-distribution in Belgium (N = 1035). The second was the information campaign on long-term radioactive waste disposal in Slovenia (N = 1,200). In both cases, recurrent and intensive communication campaigns were carried out by the authorities aiming, among other things, at increasing specific audience knowledge. Results show that higher prior audience knowledge leads to more willingness to accept communicated messages, but it does not affect people’s perception of the specific risk communicated. In addition, the influence of prior audience knowledge on the acceptance of communicated messages is shown to be no stronger than that of general radiation risk perception. The results in both case studies suggest that effective risk communication has to focus not only on knowledge but also on other more heuristic predictors, such as risk perception or attitudes toward communicated risks.

  13. Rattling Nucleons: New Developments in Active Interrogation of Special Nuclear Material

    SciTech Connect

    Robert C. Runkle; David L. Chichester; Scott J. Thompson

    2012-01-01

    Active interrogation is a vigorous area of research and development due to its promise of offering detection and characterization capabilities of special nuclear material in environments where passive detection fails. The primary value added by active methods is the capability to penetrate shielding - special nuclear material itself, incidental materials, or intentional shielding - and advocates hope that active interrogation will provide a solution to the problem of detecting shielded uranium, which is at present the greatest obstacle to interdiction efforts. The technique also provides a unique benefit for quantifying nuclear material in high background-radiation environments, an area important for nuclear material safeguards and material accountancy. Progress has been made in the field of active interrogation on several fronts, most notably in the arenas of source development, systems integration, and the integration and exploitation of multiple fission and non-fission signatures. But penetration of interrogating radiation often comes at a cost, not only in terms of finance and dose but also in terms of induced backgrounds, system complexity, and extended measurement times (including set up and acquisition). These costs make the calculus for deciding to implement active interrogation more subtle than may be apparent. The purpose of this review is thus to examine existing interrogation methods, compare and contrast their attributes and limitations, and identify missions where active interrogation may hold the most promise.

  14. Importance of the pharmacological profile of the bound ligand in enrichment on nuclear receptors: toward the use of experimentally validated decoy ligands.

    PubMed

    Lagarde, Nathalie; Zagury, Jean-François; Montes, Matthieu

    2014-10-27

    The evaluation of virtual ligand screening methods is of major importance to ensure their reliability. Taking into account the agonist/antagonist pharmacological profile should improve the quality of the benchmarking data sets since ligand binding can induce conformational changes in the nuclear receptor structure and such changes may vary according to the agonist/antagonist ligand profile. We indeed found that splitting the agonist and antagonist ligands into two separate data sets for a given nuclear receptor target significantly enhances the quality of the evaluation. The pharmacological profile of the ligand bound in the binding site of the target structure was also found to be an additional critical parameter. We also illustrate that active compound data sets for a given pharmacological activity can be used as a set of experimentally validated decoy ligands for another pharmacological activity to ensure a reliable and challenging evaluation of virtual screening methods.

  15. The link between tidal interaction and nuclear activity in galaxies

    NASA Technical Reports Server (NTRS)

    Lin, D. N. C.; Pringle, J. E.; Rees, M. J.

    1988-01-01

    It is considered how nuclear activity in galaxies may be induced by the tidal perturbation of companion galaxies. It is suggested that if the central regions of the galaxies contain marginally self-gravitating disks of gas, trailing spiral density waves, triggered by nonaxisymmetric gravitational instability, lead to efficient angular momentum transport. If the net effect of the external perturbation is to increase the effect of self-gravity in the gas, then the result is to induce a considerable increase in the mass accretion rate into the central region on a relatively short time scale. With a simple prescription, the evolution of self-gravitating accretion disks is examined in this context. These results are discussed in the context of the frequent occurrence of nuclear activity in interacting galaxies.

  16. Materials for Active Engagement in Nuclear and Particle Physics Courses

    NASA Astrophysics Data System (ADS)

    Loats, Jeff; Schwarz, Cindy; Krane, Ken

    2013-04-01

    Physics education researchers have developed a rich variety of research-based instructional strategies that now permeate many introductory courses. Carrying these active-engagement techniques to upper-division courses requires effort and is bolstered by experience. Instructors interested in these methods thus face a large investment of time to start from scratch. This NSF-TUES grant, aims to develop, test and disseminate active-engagement materials for nuclear and particle physics topics. We will present examples of these materials, including: a) Conceptual discussion questions for use with Peer Instruction; b) warm-up questions for use with Just in Time Teaching, c) ``Back of the Envelope'' estimation questions and small-group case studies that will incorporate use of nuclear and particle databases, as well as d) conceptual exam questions.

  17. Robustness of nuclear core activity reconstruction by data assimilation

    NASA Astrophysics Data System (ADS)

    Bouriquet, Bertrand; Argaud, Jean-Philippe; Erhard, Patrick; Massart, Sébastien; Ponçot, Angélique; Ricci, Sophie; Thual, Olivier

    2011-02-01

    We apply a data assimilation technique, inspired from meteorological applications, to perform an optimal reconstruction of the neutronic activity field in a nuclear core. Both measurements and information coming from a numerical model are used. We first study the robustness of the method when the amount of measured information decreases. We then study the influence of the nature of the instruments and their spatial repartition on the efficiency of the field reconstruction.

  18. Optical spectrophotometry of the nuclear region of M51. II - Further evidence for nuclear activity

    NASA Astrophysics Data System (ADS)

    Rose, J. A.; Cecil, G.

    1983-03-01

    Spectrophotometric observations of the spiral galaxy M51 conducted by Rose and Searle (1982) have revealed that the ionized gas within the central region exhibits a peculiar emission-line spectrum and is undergoing large chaotic motions. These phenomena appear to result from low-level nuclear activity qualitatively similar to that seen in Seyfert galaxy nuclei and QSOs. It has been proposed that the gas is photoionized by a central nonstellar ultraviolet continuum. The present study is concerned with a further investigation of the ionization source in the nuclear region of M51, taking into account high signal-to-noise spectra obtained with an intensified Reticon detector on the 2.24 m telescope at the Mauna Kea Observatory. It is found that photoionization by a central nonstellar ionizing continuum source provides the most consistent explanation for the observed anomalous emission-line spectrum.

  19. Importin beta1 mediates the glucose-stimulated nuclear import of pancreatic and duodenal homeobox-1 in pancreatic islet beta-cells (MIN6).

    PubMed Central

    Guillemain, Ghislaine; Da Silva Xavier, Gabriela; Rafiq, Imran; Leturque, Armelle; Rutter, Guy A

    2004-01-01

    The transcription factor PDX-1 (pancreatic and duodenal homeobox-1) is essential for pancreatic development and the maintainence of expression of islet beta-cell-specific genes. In an previous study [Rafiq, Kennedy and Rutter (1998) J. Biol. Chem. 273, 23241-23247] we demonstrated that PDX-1 may be activated at elevated glucose concentrations by translocation from undefined binding sites in the cytosol and nuclear membrane into the nucleoplasm. In the present study, we show that PDX-1 interacts directly and specifically in vitro with the nuclear import receptor family member, importin beta1, and that this interaction is mediated by the PDX-1 homeodomain (amino acids 146-206). Demonstrating the functional importance of the PDX-1-importin beta1 interaction, microinjection of MIN6 beta-cells with anti-(importin beta1) antibodies blocked both the nuclear translocation of PDX-1, and the activation by glucose (30 mM versus 3 mM) of the pre-proinsulin promoter. However, treatment with extracts from pancreatic islets incubated at either low or high glucose concentrations had no impact on the ability of PDX-1 to interact with importin beta1 in vitro. Furthermore, importin beta1 also interacted with SREBP1c (sterol-regulatory-element-binding protein 1c) in vitro, and microinjection of importin beta1 antibodies blocked the activation by glucose of SREBP1c target genes. Since the subcellular distribution of SREBP1c is unaffected by glucose, these findings suggest that a redistribution of importin beta1 is unlikely to explain the glucose-stimulated nuclear uptake of PDX-1. Instead, we conclude that the uptake of PDX-1 into the nucleoplasm, as glucose concentrations increase, may be mediated by release of the factor both from sites of retention in the cytosol and from non-productive complexes with importin beta1 at the nuclear membrane. PMID:14632628

  20. Cytoplasmic poly-GA aggregates impair nuclear import of TDP-43 in C9orf72 ALS/FTLD.

    PubMed

    Khosravi, Bahram; Hartmann, Hannelore; May, Stephanie; Möhl, Christoph; Ederle, Helena; Michaelsen, Meike; Schludi, Martin H; Dormann, Dorothee; Edbauer, Dieter

    2016-12-30

    A repeat expansion in the non-coding region of C9orf72 gene is the most common mutation causing frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). Sense and antisense transcripts are translated into aggregating dipeptide repeat (DPR) proteins in all reading frames (poly-GA,-GP,-GR,-PA and -PR) through an unconventional mechanism. How these changes contribute to cytoplasmic mislocalization and aggregation of TDP-43 and thereby ultimately lead to neuron loss remains unclear. The repeat RNA itself and poly-GR/PR have been linked to impaired nucleocytoplasmic transport. Here, we show that compact cytoplasmic poly-GA aggregates impair nuclear import of a reporter containing the TDP-43 nuclear localization (NLS) signal. However, a reporter containing a non-classical PY-NLS was not affected. Moreover, poly-GA expression prevents TNFα induced nuclear translocation of p65 suggesting that poly-GA predominantly impairs the importin-α/β-dependent pathway. In neurons, prolonged poly-GA expression induces partial mislocalization of TDP-43 into cytoplasmic granules. Rerouting poly-GA to the nucleus prevented TDP-43 mislocalization, suggesting a cytoplasmic mechanism. In rescue experiments, expression of importin-α (KPNA3, KPNA4) or nucleoporins (NUP54, NUP62) restores the nuclear localization of the TDP reporter. Taken together, inhibition of nuclear import of TDP-43 by cytoplasmic poly-GA inclusions causally links the two main aggregating proteins in C9orf72 ALS/FTLD pathogenesis.

  1. Parent participation plays an important part in promoting physical activity

    PubMed Central

    Lindqvist, Anna-Karin; Kostenius, Catrine; Gard, Gunvor; Rutberg, Stina

    2015-01-01

    Although physical activity (PA) is an important and modifiable determinant of health, in Sweden only 15% of boys and 10% of girls aged 15 years old achieve the recommended levels of PA 7 days per week. Adolescents’ PA levels are associated with social influence exerted by parents, friends, and teachers. The purpose of this study was to describe parents’ experiences of being a part of their adolescents’ empowerment-inspired PA intervention. A qualitative interview study was performed at a school in the northern part of Sweden. A total of 10 parents were interviewed, and the collected data were analyzed with qualitative content analysis. Three subthemes were combined into one main theme, demonstrating that parents are one important part of a successful PA intervention. The life of an adolescent has many options and demands that make it difficult to prioritize PA. Although parents felt that they were important in supporting their adolescent, a successful PA intervention must have multiple components. Moreover, the parents noted that the intervention had a positive effect upon not only their adolescents’, but also their own PA. Interventions aimed at promoting PA among adolescents should include measures to stimulate parent participation, have an empowerment approach, and preferably be school-based. PMID:26282870

  2. Parent participation plays an important part in promoting physical activity.

    PubMed

    Lindqvist, Anna-Karin; Kostenius, Catrine; Gard, Gunvor; Rutberg, Stina

    2015-01-01

    Although physical activity (PA) is an important and modifiable determinant of health, in Sweden only 15% of boys and 10% of girls aged 15 years old achieve the recommended levels of PA 7 days per week. Adolescents' PA levels are associated with social influence exerted by parents, friends, and teachers. The purpose of this study was to describe parents' experiences of being a part of their adolescents' empowerment-inspired PA intervention. A qualitative interview study was performed at a school in the northern part of Sweden. A total of 10 parents were interviewed, and the collected data were analyzed with qualitative content analysis. Three subthemes were combined into one main theme, demonstrating that parents are one important part of a successful PA intervention. The life of an adolescent has many options and demands that make it difficult to prioritize PA. Although parents felt that they were important in supporting their adolescent, a successful PA intervention must have multiple components. Moreover, the parents noted that the intervention had a positive effect upon not only their adolescents', but also their own PA. Interventions aimed at promoting PA among adolescents should include measures to stimulate parent participation, have an empowerment approach, and preferably be school-based.

  3. Stellar Activity and CMEs: Important Factors of Planetary Evolution

    NASA Astrophysics Data System (ADS)

    Khodachenko, Maxim L.

    CME activity of the Sun is known to be an important impacting factor for the magnetospheres, atmospheres, and surfaces of solar system planets. Following an idea of a solar-stellar analogy, CME phenomena are expected on other stars as well. The main planetary impact factors of the stellar CMEs include the associated interplanetary shocks, plasma density and velocity disturbances, energetic particles accelerated in the shock regions, as well as distortions of the magnetic field direction and modulus. All these factors should be properly taken into account during the study of evolutionary processes on exoplanets and their atmospheric and plasma environments. The planetary impact of the stellar CME activity may vary depending on stellar age, stellar spectral type and the orbital distance of a planet. Because of the relatively short range of propagation of the majority of CMEs, they affect most strongly the magnetospheres and atmospheres of close-orbit ( < 0.1 AU) exoplanets. In this chapter we discuss an issue of the stellar CME activity in the context of several actual problems of modern exoplanetology, including planetary atmosphere mass loss, planet survival at close orbits, and definition of a criterion for habitability.

  4. Charged fusion product loss measurements using nuclear activation

    SciTech Connect

    Bonheure, G.; Hult, M.; Gonzalez de Orduna, R.; Wieslander, E.; Arnold, D.; Dombrowski, H.; Laubenstein, M.; Murari, A.; Collaboration: JET-EFDA Contributors

    2010-10-15

    In ITER, {alpha} particle loss measurements will be required in order to understand the alpha particle physics. Techniques capable of operating in a fusion reactor environment need further development. Recent experimental studies on JET demonstrated the potential of nuclear activation to measure the flux of escaping MeV ions. New results from MeV ion induced activation of metallic, ceramic, and crystal samples placed near the plasma edge are reported. Activation products were measured as function of orientation with respect to the magnetic field as well as function of the distance to the plasma. Sample activity was measured using ultralow-level gamma-ray spectrometry. Distribution of 14.68 MeV fusion proton induced activation products is strongly anisotropic in agreement with simulations and falls off sharply with increasing distance to the plasma. Prospects for using the technique in ITER are discussed.

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

    PubMed Central

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

    2015-01-01

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

  6. Determination of Important Nuclear Fragmentation Processes for Human Space Radiation Protection

    NASA Technical Reports Server (NTRS)

    Lin, Zi-Wei

    2007-01-01

    We present a semi-analytical method to determine which partial cross sections of nuclear fragmentations most affect the shielded dose equivalent due to exposure to galactic cosmic rays. The cross sections thus determined will require more theoretical and/or experimental studies in order for us to better predict, reduce and mitigate the radiation exposure in human space explorations.

  7. The importance of non-nuclear AR signaling in prostate cancer progression and therapeutic resistance.

    PubMed

    Zarif, Jelani C; Miranti, Cindy K

    2016-05-01

    The androgen receptor (AR) remains the major oncogenic driver of prostate cancer, as evidenced by the efficacy of androgen deprivation therapy (ADT) in naïve patients, and the continued effectiveness of second generation ADTs in castration resistant disease. However, current ADTs are limited to interfering with AR ligand binding, either through suppression of androgen production or the use of competitive antagonists. Recent studies demonstrate 1) the expression of constitutively active AR splice variants that no longer depend on androgen, and 2) the ability of AR to signal in the cytoplasm independently of its transcriptional activity (non-genomic); thus highlighting the need to consider other ways to target AR. Herein, we review canonical AR signaling, but focus on AR non-genomic signaling, some of its downstream targets and how these effectors contribute to prostate cancer cell behavior. The goals of this review are to 1) re-highlight the continued importance of AR in prostate cancer as the primary driver, 2) discuss the limitations in continuing to use ligand binding as the sole targeting mechanism, 3) discuss the implications of AR non-genomic signaling in cancer progression and therapeutic resistance, and 4) address the need to consider non-genomic AR signaling mechanisms and pathways as a viable targeting strategy in combination with current therapies.

  8. [Occupational radiation exposures during maintenance activities at nuclear power plants].

    PubMed

    Imahori, A

    1987-11-01

    Occupational exposures at nuclear power plants occur mostly during maintenance activities rather than during routine reactor operation. In this paper, statistical summaries of occupational exposures during routine maintenance activities for the years 1982-84 at nuclear power plants in Japan are presented, including comparison of the exposure levels by reactor type and by plant age. Average annual collective doses per reactor for BWRs and PWRs are 7.30 man-Sv and 2.84 man-Sv, respectively, and 78% and 89% of annual doses are incurred during maintenance activities. Average annual outage days of BWRs and PWRs for routine maintenance are 102 d and 97 d. Annual collective doses per reactor, most of which occur during maintenance activities, usually increase with plant age. Higher collective doses are observed for routine maintenance performed on older reactors as compared to newer reactors, especially in BWRs. Collective doses accrued during respective routine maintenance activities have a significant correlation with duration of maintenance and number of workers involved in maintenance.

  9. Pakistan’s Nuclear Proliferation Activities and the Recommendations of the 9/11 Commission: U.S. Policy Constraints and Options

    DTIC Science & Technology

    2005-03-16

    Pakistan no longer critical to U.S. regional security policy.15 India and Pakistan’s May 1998 Nuclear Tests and the Decline of Sanctions as a U.S... nuclear activities. A number of nonproliferation experts agree with the critical importance of keeping Pakistan’s nuclear weapons and materials out of...plans for an uranium enrichment facility, this is only one — albeit a critical one — of many steps required in the production of nuclear weapons

  10. Biological significance of nuclear localization of mitogen-activated protein kinase Pmk1 in fission yeast.

    PubMed

    Sánchez-Mir, Laura; Franco, Alejandro; Madrid, Marisa; Vicente-Soler, Jero; Villar-Tajadura, M Antonia; Soto, Teresa; Pérez, Pilar; Gacto, Mariano; Cansado, José

    2012-07-27

    Mitogen-activated protein kinase (MAPK) signaling pathways play a fundamental role in the response of eukaryotic cells to environmental changes. Also, much evidence shows that the stimulus-dependent nuclear targeting of this class of regulatory kinases is crucial for adequate regulation of distinct cellular events. In the fission yeast Schizosaccharomyces pombe, the cell integrity MAPK pathway, whose central element is the MAPK Pmk1, regulates multiple processes such as cell wall integrity, vacuole fusion, cytokinesis, and ionic homeostasis. In non-stressed cells Pmk1 is constitutively localized in both cytoplasm and nucleus, and its localization pattern appears unaffected by its activation status or in response to stress, thus questioning the biological significance of the presence of this MAPK into the nucleus. We have addressed this issue by characterizing mutants expressing Pmk1 versions excluded from the cell nucleus and anchored to the plasma membrane in different genetic backgrounds. Although nuclear Pmk1 partially regulates cell wall integrity at a transcriptional level, membrane-tethered Pmk1 performs many of the biological functions assigned to wild type MAPK like regulation of chloride homeostasis, vacuole fusion, and cellular separation. However, we found that down-regulation of nuclear Pmk1 by MAPK phosphatases induced by the stress activated protein kinase pathway is important for the fine modulation of extranuclear Pmk1 activity. These results highlight the importance of the control of MAPK activity at subcellular level.

  11. Biological Significance of Nuclear Localization of Mitogen-activated Protein Kinase Pmk1 in Fission Yeast*

    PubMed Central

    Sánchez-Mir, Laura; Franco, Alejandro; Madrid, Marisa; Vicente-Soler, Jero; Villar-Tajadura, M. Antonia; Soto, Teresa; Pérez, Pilar; Gacto, Mariano; Cansado, José

    2012-01-01

    Mitogen-activated protein kinase (MAPK) signaling pathways play a fundamental role in the response of eukaryotic cells to environmental changes. Also, much evidence shows that the stimulus-dependent nuclear targeting of this class of regulatory kinases is crucial for adequate regulation of distinct cellular events. In the fission yeast Schizosaccharomyces pombe, the cell integrity MAPK pathway, whose central element is the MAPK Pmk1, regulates multiple processes such as cell wall integrity, vacuole fusion, cytokinesis, and ionic homeostasis. In non-stressed cells Pmk1 is constitutively localized in both cytoplasm and nucleus, and its localization pattern appears unaffected by its activation status or in response to stress, thus questioning the biological significance of the presence of this MAPK into the nucleus. We have addressed this issue by characterizing mutants expressing Pmk1 versions excluded from the cell nucleus and anchored to the plasma membrane in different genetic backgrounds. Although nuclear Pmk1 partially regulates cell wall integrity at a transcriptional level, membrane-tethered Pmk1 performs many of the biological functions assigned to wild type MAPK like regulation of chloride homeostasis, vacuole fusion, and cellular separation. However, we found that down-regulation of nuclear Pmk1 by MAPK phosphatases induced by the stress activated protein kinase pathway is important for the fine modulation of extranuclear Pmk1 activity. These results highlight the importance of the control of MAPK activity at subcellular level. PMID:22685296

  12. Nuclear import of cutaneous beta genus HPV8 E7 oncoprotein is mediated by hydrophobic interactions between its zinc-binding domain and FG nucleoporins

    SciTech Connect

    Onder, Zeynep; Moroianu, Junona

    2014-01-20

    We have previously discovered and characterized the nuclear import pathways for the E7 oncoproteins of mucosal alpha genus HPVs, type 16 and 11. Here we investigated the nuclear import of cutaneous beta genus HPV8 E7 protein using confocal microscopy after transfections of HeLa cells with EGFP-8E7 and mutant plasmids and nuclear import assays in digitonin-permeabilized HeLa cells. We determined that HPV8 E7 contains a nuclear localization signal (NLS) within its zinc-binding domain that mediates its nuclear import. Furthermore, we discovered that a mostly hydrophobic patch {sub 65}LRLFV{sub 69} within the zinc-binding domain is essential for the nuclear import and localization of HPV8 E7 via hydrophobic interactions with the FG nucleoporins Nup62 and Nup153. Substitution of the hydrophobic residues within the {sub 65}LRLFV{sub 69} patch to alanines, and not R66A mutation, disrupt the interactions between the 8E7 zinc-binding domain and Nup62 and Nup153 and consequently inhibit nuclear import of HPV8 E7. - Highlights: • HPV8 E7 has a cNLS within its zinc-binding domain that mediates its nuclear import. • Discovery of a hydrophobic patch that is critical for the nuclear import of HPV8 E7. • HPV8 E7 nuclear import is mediated by hydrophobic interactions with FG-Nups, Nup62 and Nup153.

  13. Two domains of the V protein of virulent canine distemper virus selectively inhibit STAT1 and STAT2 nuclear import.

    PubMed

    Röthlisberger, Anne; Wiener, Dominique; Schweizer, Matthias; Peterhans, Ernst; Zurbriggen, Andreas; Plattet, Philippe

    2010-07-01

    Canine distemper virus (CDV) causes in dogs a severe systemic infection, with a high frequency of demyelinating encephalitis. Among the six genes transcribed by CDV, the P gene encodes the polymerase cofactor protein (P) as well as two additional nonstructural proteins, C and V; of these V was shown to act as a virulence factor. We investigated the molecular mechanisms by which the P gene products of the neurovirulent CDV A75/17 strain disrupt type I interferon (IFN-alpha/beta)-induced signaling that results in the establishment of the antiviral state. Using recombinant knockout A75/17 viruses, the V protein was identified as the main antagonist of IFN-alpha/beta-mediated signaling. Importantly, immunofluorescence analysis illustrated that the inhibition of IFN-alpha/beta-mediated signaling correlated with impaired STAT1/STAT2 nuclear import, whereas the phosphorylation state of these proteins was not affected. Coimmunoprecipitation assays identified the N-terminal region of V (VNT) responsible for STAT1 targeting, which correlated with its ability to inhibit the activity of the IFN-alpha/beta-mediated antiviral state. Conversely, while the C-terminal domain of V (VCT) could not function autonomously, when fused to VNT it optimally interacted with STAT2 and subsequently efficiently suppressed the IFN-alpha/beta-mediated signaling pathway. The latter result was further supported by a single mutation at position 110 within the VNT domain of CDV V protein, resulting in a mutant that lost STAT1 binding while retaining a partial STAT2 association. Taken together, our results identified the CDV VNT and VCT as two essential modules that complement each other to interfere with the antiviral state induced by IFN-alpha/beta-mediated signaling. Hence, our experiments reveal a novel mechanism of IFN-alpha/beta evasion among the morbilliviruses.

  14. Nucleoporins NPP-10, NPP-13 and NPP-20 are required for HCP-4 nuclear import to establish correct centromere assembly.

    PubMed

    Ferreira, Jorge; Stear, Jeffrey H; Saumweber, Harald

    2017-03-01

    Centromeres form a chromosomal platform for the assembly of the kinetochores, which are required for orderly chromosome segregation. Assembly of both centromeres and kinetochores proceeds by a step-by-step mechanism that is regulated in time and space. It has been suggested that the regulated nuclear import of centromeric proteins is involved in this process. We show that the knockdown of nucleoporins NPP-10, NPP-13 and NPP-20 in Caenorhabditiselegans affects early steps in centromere formation and sister centromere resolution, and results in severe chromosomal defects in the early embryo. These phenotypes mirror the knockdown phenotype of HCP-4 (an ortholog of mammalian CENP-C), a key factor for centromere formation and inner kinetochore assembly. HCP-4 is present in the cytoplasm during interphase. It is imported into nuclei and assembled in centromeres during prophase. Following the knockdown of NPP-10, NPP-13 and NPP-20, HCP-4 remains in the cytosol throughout prophase due to stalled import. In prometaphase and later mitotic stages after breakdown of the nuclear envelope, HCP-4 is not incorporated into centromeres. These results indicate that correct timing of the availability of HCP-4 by nuclear import is essential.

  15. The importance of the AVR pebble-bed reactor for the future of nuclear power

    SciTech Connect

    Pohl, P.

    2006-07-01

    The AVR pebble-bed high temperature gas-cooled reactor (HTGR) at Juelich (Germany)) operated from 1967 to 1988 and was certainly the most important HTGR project of the past. The reactor was the mass test bed for all development steps of HTGR pebble fuel. Some early fuel charges failed under high temperature conditions and contaminated the reactor. An accurate pebble measurement (Cs 137) allowed to clean the core from unwanted pebbles after 1981. The coolant activity went down and remained very low for the remaining reactor operation. A melt-wire experiment in 1986 revealed max. coolant temperatures of >1280 deg. C and fuel temperatures of >1350 deg. C, explained by under-estimated bypasses. The fuel still in the core achieved high burn-ups and showed under the extreme temperature conditions excellent fission product retention. Thus, the AVR operation qualified the HTGR fuel, and an average discharge burn-up of 112% fifa revealed an excellent fuel economy of the pebble-bed reactor. Furthermore, the AVR operation offers many meaningful data for code-to-experiment comparisons. (authors)

  16. Arenavirus Nucleoproteins Prevent Activation of Nuclear Factor Kappa B

    PubMed Central

    Rodrigo, W. W. Shanaka I.; Ortiz-Riaño, Emilio; Pythoud, Christelle; Kunz, Stefan

    2012-01-01

    Arenaviruses include several causative agents of hemorrhagic fever (HF) disease in humans that are associated with high morbidity and significant mortality. Morbidity and lethality associated with HF arenaviruses are believed to involve the dysregulation of the host innate immune and inflammatory responses that leads to impaired development of protective and efficient immunity. The molecular mechanisms underlying this dysregulation are not completely understood, but it is suggested that viral infection leads to disruption of early host defenses and contributes to arenavirus pathogenesis in humans. We demonstrate in the accompanying paper that the prototype member in the family, lymphocytic choriomeningitis virus (LCMV), disables the host innate defense by interfering with type I interferon (IFN-I) production through inhibition of the interferon regulatory factor 3 (IRF3) activation pathway and that the viral nucleoprotein (NP) alone is responsible for this inhibitory effect (C. Pythoud, W. W. Rodrigo, G. Pasqual, S. Rothenberger, L. Martínez-Sobrido, J. C. de la Torre, and S. Kunz, J. Virol. 86:7728–7738, 2012). In this report, we show that LCMV-NP, as well as NPs encoded by representative members of both Old World (OW) and New World (NW) arenaviruses, also inhibits the nuclear translocation and transcriptional activity of the nuclear factor kappa B (NF-κB). Similar to the situation previously reported for IRF3, Tacaribe virus NP (TCRV-NP) does not inhibit NF-κB nuclear translocation and transcriptional activity to levels comparable to those seen with other members in the family. Altogether, our findings demonstrate that arenavirus infection inhibits NF-κB-dependent innate immune and inflammatory responses, possibly playing a key role in the pathogenesis and virulence of arenavirus. PMID:22623788

  17. Arenavirus nucleoproteins prevent activation of nuclear factor kappa B.

    PubMed

    Rodrigo, W W Shanaka I; Ortiz-Riaño, Emilio; Pythoud, Christelle; Kunz, Stefan; de la Torre, Juan C; Martínez-Sobrido, Luis

    2012-08-01

    Arenaviruses include several causative agents of hemorrhagic fever (HF) disease in humans that are associated with high morbidity and significant mortality. Morbidity and lethality associated with HF arenaviruses are believed to involve the dysregulation of the host innate immune and inflammatory responses that leads to impaired development of protective and efficient immunity. The molecular mechanisms underlying this dysregulation are not completely understood, but it is suggested that viral infection leads to disruption of early host defenses and contributes to arenavirus pathogenesis in humans. We demonstrate in the accompanying paper that the prototype member in the family, lymphocytic choriomeningitis virus (LCMV), disables the host innate defense by interfering with type I interferon (IFN-I) production through inhibition of the interferon regulatory factor 3 (IRF3) activation pathway and that the viral nucleoprotein (NP) alone is responsible for this inhibitory effect (C. Pythoud, W. W. Rodrigo, G. Pasqual, S. Rothenberger, L. Martínez-Sobrido, J. C. de la Torre, and S. Kunz, J. Virol. 86:7728-7738, 2012). In this report, we show that LCMV-NP, as well as NPs encoded by representative members of both Old World (OW) and New World (NW) arenaviruses, also inhibits the nuclear translocation and transcriptional activity of the nuclear factor kappa B (NF-κB). Similar to the situation previously reported for IRF3, Tacaribe virus NP (TCRV-NP) does not inhibit NF-κB nuclear translocation and transcriptional activity to levels comparable to those seen with other members in the family. Altogether, our findings demonstrate that arenavirus infection inhibits NF-κB-dependent innate immune and inflammatory responses, possibly playing a key role in the pathogenesis and virulence of arenavirus.

  18. Department of Energy interest and involvement in nuclear plant license renewal activities

    SciTech Connect

    Bustard, L.D. ); Harrison, D.L. . Office of LWR Safety and Technology)

    1991-01-01

    Recognizing the importance of nuclear license renewal to the nation's energy strategy, the Department of Energy (DOE) initiated a plant lifetime improvement program during 1985 to determine the feasibility of the license renewal option for US nuclear plants. Initial activities of the DOE program focused on determining whether there were technical and economic obstacles that might preclude or limit the successful implementation of the license renewal option. To make this determination, DOE cosponsored with the Electric Power Research Institute (EPRI) pilot-plant efforts by Virginia Electric Power and Northern States Power. Both pilot-plant efforts concluded that life extension is technically and economically feasible. In parallel with the pilot-plant activities, DOE performed national economic studies that demonstrated the economic desirability of life extension. Having demonstrated the feasibility of life extension, DOE, in conjunction with EPRI, selected two lead plants to demonstrate the license renewal process. These lead plants are Yankee Atomic's Yankee Rowe facility and Northern States Power's Monticello facility. DOE also initiated activities to develop the technical and regulatory bases to support the license renewal process in the United States. DOE has recently identified nuclear plant license renewal to be an important element of its National Energy Strategy. This paper summarizes the significant results, conclusions, and ongoing activities of the DOE effort. 18 refs.

  19. Nuclear Physics Activities in Asia and ANPhA

    SciTech Connect

    Sakai, H.

    2011-05-06

    On 18 July 2009 the Asian Nuclear Physics Association (ANPhA) has been officially launched in Beijing by the representatives from China, Korea, Japan and Vietnam. Since then Australia, India, Mongolia and Taiwan have joined to ANPhA and now the member country/region has increased to eight. Some activities and features on ANPhA are introduced. In addition, pleasant collaboration with Professor Arima by the author in regard to the Gamow-Teller quenching problem is also briefly mentioned.

  20. Determine Important Nuclear Fragmentation Processes for Space Radiation Protection in Human Space Explorations

    NASA Technical Reports Server (NTRS)

    Lin, Zi-Wei

    2004-01-01

    Space radiation from cosmic ray particles is one of the main challenges for long-term human space explorations such as a permanent moon base or a trip to Mars. Material shielding may provide significant radiation protection to astronauts, and models have been developed in order to evaluate the effectiveness of different shielding materials and to predict radiation environment inside the spacecraft. In this study we determine the nuclear fragmentation cross sections which will most affect the radiation risk behind typical radiation shielding materials. These cross sections thus need more theoretical studies and accurate experimental measurements in order for us to more precisely predict the radiation risk in human space exploration.

  1. Determine Important Nuclear Fragmentation Processes for Space Radiation Protection in Human Space Explorations

    NASA Technical Reports Server (NTRS)

    Lin, Zi-wei

    2004-01-01

    Space radiation from cosmic ray particles is one of the main challenges for long-term human space explorations such as a permanent moon base or a trip to Mars. Material shielding may provide significant radiation protection to astronauts, and models have been developed in order to evaluate the effectiveness of different shielding materials and to predict radiation environment inside the spacecraft. In this study we determine the nuclear fragmentation cross sections which will most effect the radiation risk behind typical radiation shielding materials. These cross sections thus need more theoretical studies and accurate experimental measurements in order for us to more precisely predict the radiation risk in human space explorations.

  2. Determine Important Nuclear Fragmentation Processes for Space Radiation Protection in Human Space Explorations

    NASA Technical Reports Server (NTRS)

    Lin, Zi-Wei

    2004-01-01

    Space radiation from cosmic ray particles is one of the main challenges for long-term human space explorations such as a permanent moon base or a trip to Mars. Material shielding may provide significant radiation protection to astronauts, and models have been developed in order to evaluate the effectiveness of different shielding materials and to predict radiation environment inside the spacecraft. In this study we determine the nuclear fragmentation cross sections which will most affect the radiation risk behind typical radiation shielding materials. These cross sections thus need more theoretical studies and accurate experimental measurements in order for us to more precisely predict the radiation risk in human space explorations.

  3. Nuclear import of prototype foamy virus transactivator Bel1 is mediated by KPNA1, KPNA6 and KPNA7.

    PubMed

    Duan, Jihui; Tang, Zhiqin; Mu, Hong; Zhang, Guojun

    2016-08-01

    Bel1, a transactivator of the prototype foamy virus (PFV), plays pivotal roles in the replication of PFV. Previous studies have demonstrated that Bel1 bears a nuclear localization signal (NLS); however, its amino acid sequence remains unclear and the corresponding adaptor importins have not yet been identified. In this study, we inserted various fragments of Bel1 into an EGFP-GST fusion protein and investigated their subcellular localization by fluorescence microscopy. We found that the 215PRQKRPR221 fragment, which accords with the consensus sequence K(K/R)X(K/R) of the monopartite NLS, directed the nuclear translocation of Bel1. Point mutation experiments revealed that K218, R219 and R221 were essential for the nuclear localization of Bel1. The results of GST pull-down assay revealed that the Bel1 peptide 215-221, which bears the NLS, interacted with the nucleocytoplasmic transport receptors, karyopherin alpha 1 (importin alpha 5) (KPNA1), karyopherin alpha 6 (importin alpha 7) (KPNA6) and karyopherin alpha 7 (importin alpha 8) (KPNA7). Finally, in vitro nuclear import assays demonstrated that KPNA1, KPNA6 or KPNA7, along with other necessary nuclear factors, caused Bel1 to localize to the nucleus. Thus, the findings of our study indicate that KPNA1, KPNA6 and KPNA7 are involved in Bel1 nuclear distribution.

  4. Lessons Learned from the Decommissioning of Nuclear Facilities and the Safe Termination of Nuclear Activities. Outcomes of the International Conference, 11-15 December 2006, Athens, Greece

    SciTech Connect

    Batandjieva, B.; Laraia, M.

    2008-01-15

    Full text of publication follows: decommissioning activities are increasing worldwide covering wide range of facilities - from nuclear power plant, through fuel cycle facilities to small laboratories. The importance of these activities is growing with the recognition of the need for ensuring safe termination of practices and reuse of sites for various purposes, including the development of new nuclear facilities. Decommissioning has been undertaken for more than forty years and significant knowledge has been accumulated and lessons have been learned. However the number of countries encountering decommissioning for the first time is increasing with the end of the lifetime of the facilities around the world, in particular in countries with small nuclear programmes (e.g. one research reactor) and limited human and financial resources. In order to facilitate the exchange of lessons learned and good practices between all Member States and to facilitate and improve safety of the planned, ongoing and future decommissioning projects, the IAEA in cooperation with the Nuclear Energy Agency to OECD, European Commission and World Nuclear Association organised the international conference on Lessons Learned from the Decommissioning of Nuclear Facilities and the Safe Termination of Nuclear Activities, held in Athens, Greece. The conference also highlighted areas where future cooperation at national and international level is required in order to improve decommissioning planning and safety during decommissioning and to facilitate decommissioning by selecting appropriate strategies and technologies for decontamination, dismantling and management of waste. These and other aspects discussed at the conference are presented in this paper, together with the planned IAEA measures for amendment and implementation of the International Action Plan on Decommissioning of Nuclear Facilities and its future programme on decommissioning.

  5. The Nuclear Accident at Three Mile Island a Practical Lesson in the Fundamental Importance of Effective Communications

    SciTech Connect

    DeVine Jr, J.C.

    2008-07-01

    The Three Mile Island Unit 2 (TMI-2) accident in March 1979 had a profound effect on the course of commercial nuclear generation in the United States and around the world. And while the central elements of the accident were matters of nuclear engineering, design and operations, its consequences were compounded, and in some respects superseded, by extraordinarily ineffective communications by all parties at all levels. Communications failures during the accident and its aftermath caused misunderstanding, distrust, and incorrect emergency response - and seeded or reinforced public opposition to nuclear power that persists to this day. There are communications lessons from TMI that have not yet been fully learned, and some that once were learned but are now gradually being forgotten. The more glaring TMI communications problems were in the arena of external interactions and communications among the plant owner, the Nuclear Regulatory Commission (NRC), the media, and the public. Confusing, fragmented, and contradictory public statements early in the accident, regardless of cause, undermined all possibility for reasonable discourse thereafter. And because the TMI accident was playing out on a world stage, the breakdown in public trust had long term and widespread implications. At the plant site, both TMI-2 cleanup and restart of the undamaged TMI-1 unit met with years of public and political criticism, and attendant regulatory pressure. Across the nation, public trust in nuclear power and those who operate it plummeted, unquestionably contributing to the 25+ year hiatus in new plant orders. There were other, less visible but equally important, consequences of ineffective communications at TMI. The unplanned 'precautionary' evacuation urged by the governor two days after the accident - a life changing, traumatic event for thousands of residents - was prompted primarily by misunderstandings and miscommunications regarding the condition of the plant. And today, nearly 30

  6. Identification of COUP-TFII Orphan Nuclear Receptor as a Retinoic Acid–Activated Receptor

    PubMed Central

    Kruse, Schoen W; Suino-Powell, Kelly; Zhou, X. Edward; Kretschman, Jennifer E; Reynolds, Ross; Vonrhein, Clemens; Xu, Yong; Wang, Liliang; Tsai, Sophia Y; Tsai, Ming-Jer; Xu, H. Eric

    2008-01-01

    The chicken ovalbumin upstream promoter-transcription factors (COUP-TFI and II) make up the most conserved subfamily of nuclear receptors that play key roles in angiogenesis, neuronal development, organogenesis, cell fate determination, and metabolic homeostasis. Although the biological functions of COUP-TFs have been studied extensively, little is known of their structural features or aspects of ligand regulation. Here we report the ligand-free 1.48 Å crystal structure of the human COUP-TFII ligand-binding domain. The structure reveals an autorepressed conformation of the receptor, where helix α10 is bent into the ligand-binding pocket and the activation function-2 helix is folded into the cofactor binding site, thus preventing the recruitment of coactivators. In contrast, in multiple cell lines, COUP-TFII exhibits constitutive transcriptional activity, which can be further potentiated by nuclear receptor coactivators. Mutations designed to disrupt cofactor binding, dimerization, and ligand binding, substantially reduce the COUP-TFII transcriptional activity. Importantly, retinoid acids are able to promote COUP-TFII to recruit coactivators and activate a COUP-TF reporter construct. Although the concentration needed is higher than the physiological levels of retinoic acids, these findings demonstrate that COUP-TFII is a ligand-regulated nuclear receptor, in which ligands activate the receptor by releasing it from the autorepressed conformation. PMID:18798693

  7. Identification of COUP-TFII Orphan Nuclear Receptor as a Retinoic Acid-Activated Receptor

    SciTech Connect

    Kruse, Schoen W; Suino-Powell, Kelly; Zhou, X Edward; Kretschman, Jennifer E; Reynolds, Ross; Vonrhein, Clemens; Xu, Yong; Wang, Liliang; Tsai, Sophia Y; Tsai, Ming-Jer; Xu, H Eric

    2010-01-12

    The chicken ovalbumin upstream promoter-transcription factors (COUP-TFI and II) make up the most conserved subfamily of nuclear receptors that play key roles in angiogenesis, neuronal development, organogenesis, cell fate determination, and metabolic homeostasis. Although the biological functions of COUP-TFs have been studied extensively, little is known of their structural features or aspects of ligand regulation. Here we report the ligand-free 1.48 {angstrom} crystal structure of the human COUP-TFII ligand-binding domain. The structure reveals an autorepressed conformation of the receptor, where helix {alpha}10 is bent into the ligand-binding pocket and the activation function-2 helix is folded into the cofactor binding site, thus preventing the recruitment of coactivators. In contrast, in multiple cell lines, COUP-TFII exhibits constitutive transcriptional activity, which can be further potentiated by nuclear receptor coactivators. Mutations designed to disrupt cofactor binding, dimerization, and ligand binding, substantially reduce the COUP-TFII transcriptional activity. Importantly, retinoid acids are able to promote COUP-TFII to recruit coactivators and activate a COUP-TF reporter construct. Although the concentration needed is higher than the physiological levels of retinoic acids, these findings demonstrate that COUP-TFII is a ligand-regulated nuclear receptor, in which ligands activate the receptor by releasing it from the autorepressed conformation.

  8. Comparison of Different Internal Dosimetry Systems for Selected Radionuclides Important to Nuclear Power Production

    SciTech Connect

    Leggett, Richard Wayne; Eckerman, Keith F; Manger, Ryan P

    2013-08-01

    This report compares three different radiation dosimetry systems currently applied by various U.S. Federal agencies and dose estimates based on these three dosimetry systems for a set of radionuclides often identified in power reactor effluents. These dosimetry systems were developed and applied by the International Commission on Radiological Protection at different times over the past six decades. Two primary modes of intake of radionuclides are addressed: ingestion in drinking water and inhalation. Estimated doses to individual organs and to the whole body based on each dosimetry system are compared for each of four age groups: infant, child, teenager, and adult. Substantial differences between dosimetry systems in estimated dose per unit intake are found for some individual radionuclides, but differences in estimated dose per unit intake generally are modest for mixtures of radionuclides typically found in nuclear power plant effluents.

  9. Active Learning: The Importance of Developing a Comprehensive Measure

    ERIC Educational Resources Information Center

    Carr, Rodney; Palmer, Stuart; Hagel, Pauline

    2015-01-01

    This article reports on an investigation into the validity of a widely used scale for measuring the extent to which higher education students employ active learning strategies. The scale is the active learning scale in the Australasian Survey of Student Engagement. This scale is based on the Active and Collaborative Learning scale of the National…

  10. 70-kDa heat shock cognate protein hsc70 mediates calmodulin-dependent nuclear import of the sex-determining factor SRY.

    PubMed

    Kaur, Gurpreet; Lieu, Kim G; Jans, David A

    2013-02-08

    We recently showed that the developmentally important family of SOX (SRY (sex determining region on the Y chromosome)-related high mobility group (HMG) box) proteins require the calcium-binding protein calmodulin (CaM) for optimal nuclear accumulation, with clinical mutations in SRY that specifically impair nuclear accumulation via this pathway resulting in XY sex reversal. However, the mechanism by which CaM facilitates nuclear accumulation is unknown. Here, we show, for the first time, that the 70-kDa heat shock cognate protein hsc70 plays a key role in CaM-dependent nuclear import of SRY. Using a reconstituted nuclear import assay, we show that antibodies to hsc70 significantly reduce nuclear accumulation of wild type SRY and mutant derivatives thereof that retain CaM-dependent nuclear import, with an increased rate of nuclear accumulation upon addition of both CaM and hsc70, in contrast to an SRY mutant derivative with impaired CaM binding. siRNA knockdown of hsc70 in intact cells showed similar results, indicating clear dependence upon hsc70 for CaM-dependent nuclear import. Analysis using the technique of fluorescence recovery after photobleaching indicated that hsc70 is required for the maximal rate of SRY nuclear import in living cells but has no impact upon SRY nuclear retention/nuclear dynamics. Finally, we demonstrate direct binding of hsc70 to the SRY·CaM complex, with immunoprecipitation experiments from cell extracts showing association of hsc70 with wild type SRY, but not with a mutant derivative with impaired CaM binding, dependent on Ca(2+). Our novel findings strongly implicate hsc70 in CaM-dependent nuclear import of SRY.

  11. Formation of Tap/NXT1 Heterodimers Activates Tap-Dependent Nuclear mRNA Export by Enhancing Recruitment to Nuclear Pore Complexes

    PubMed Central

    Wiegand, Heather L.; Coburn, Glen A.; Zeng, Yan; Kang, Yibin; Bogerd, Hal P.; Cullen, Bryan R.

    2002-01-01

    The Tap protein has been shown to activate the nuclear export of mRNA species bearing retroviral constitutive transport elements and is also believed to play an essential role in the sequence nonspecific export of cellular mRNAs. However, it has remained unclear how Tap activity is regulated in vivo. Here, we report that the small NXT1/p15-1 protein functions as a critical cofactor for Tap-mediated mRNA export in both human and invertebrate cells. In the absence of NXT1 binding, the Tap protein is unable to effectively interact with components of the nuclear pore complex and both Tap nucleocytoplasmic shuttling and the nuclear export of mRNA molecules tethered to Tap are therefore severely attenuated. Formation of a Tap/NXT1 heterodimer enhances nucleoporin binding both in vitro and in vivo and induces the formation of a Tap/NXT1/nucleoporin ternary complex that is likely to be a key intermediate in the process of nuclear mRNA export. The critical importance of NXT1 for the nuclear export of poly(A)+ RNA is emphasized by the finding that specific inhibition of the expression of the Drosophila homolog of human NXT1, by using RNA interference, results in the nuclear accumulation of poly(A)+ RNA in cultured insect cells. These data suggest that NXT1 may act as a molecular switch that regulates the ability of Tap to mediate nuclear mRNA export by controlling the interaction of Tap with components of the nuclear pore. PMID:11739738

  12. Active microrheology of a model of the nuclear micromechanical environment

    NASA Astrophysics Data System (ADS)

    Byrd, Henry; Kilfoil, Maria

    2014-03-01

    In order to successfully complete the final stages of chromosome segregation, eukaryotic cells require the motor enzyme topoisomerase II, which can resolve topological constraints between entangled strands of duplex DNA. We created an in vitro model of a close approximation of the nuclear micromechanical environment in terms of DNA mass and entanglement density, and investigated the influence of this motor enzyme on the DNA mechanics. Topoisomerase II is a non-processive ATPase which we found significantly increases the motions of embedded microspheres in the DNA network. Because of this activity, we study the mechanical properties of our model system by active microrheology by optical trapping. We test the limits of fluctuation dissipation theorem (FDT) under this type of activity by comparing the active microrheology to passive measurements, where thermal motion alone drives the beads. We can relate any departure from FDT to the timescale of topoisomerase II activity in the DNA network. These experiments provide insight into the physical necessity of this motor enzyme in the cell.

  13. Nuclear Translocation Sequence and Region in Autographa californica Multiple Nucleopolyhedrovirus ME53 That Are Important for Optimal Baculovirus Production

    PubMed Central

    Liu, Yang; de Jong, Jondavid; Nagy, Éva; Theilmann, David A.

    2016-01-01

    ABSTRACT Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is in the family Baculoviridae, genus Alphabaculovirus. AcMNPV me53 is a highly conserved immediate early gene in all lepidopteran baculoviruses that have been sequenced and is transcribed up to late times postinfection. Although me53 is not essential for viral DNA synthesis, infectious budded virus (BV) production is greatly attenuated when it is deleted. ME53 associates with the nucleocapsid on both budded virus and occlusion-derived virus, but not with the virus envelope. ME53 colocalizes in plasma membrane foci with the envelope glycoprotein GP64 in a GP64-dependent manner. ME53 localizes in the cytoplasm early postinfection, and despite the lack of a reported nuclear localization signal (NLS), ME53 translocates to the nucleus at late times postinfection. To map determinants of ME53 that facilitate its nuclear translocation, recombinant AcMNPV bacmids containing a series of ME53 truncations, internal deletions, and peptides fused with hemagglutinin (HA) or green fluorescent protein (GFP) tags were constructed. Intracellular-localization studies identified residues within amino acids 109 to 137 at the N terminus of ME53 that acted as the nuclear translocation sequence (NTS), facilitating its nuclear transport at late times postinfection. The first 100 N-terminal amino acids and the last 50 C-terminal amino acids of ME53 are dispensable for high levels of budded virus production. The region within amino acids 101 to 398, which also contains the NTS, is critical for optimal levels of budded virus production. IMPORTANCE Baculovirus me53 is a conserved immediate early gene found in all sequenced lepidopteran alpha- and betabaculoviruses. We first identified residues within amino acids 109 to 137 at the N terminus that act as the ME53 nuclear translocation sequence (NTS) to facilitate its nuclear translocation and defined an internal region within amino acids 101 to 398, which includes the NTS, as

  14. 75 FR 50772 - Agency Information Collection Activities: Importation Bond Structure

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-17

    ... Structure AGENCY: U.S. Customs and Border Protection, Department of Homeland Security. ] ACTION: 60-Day... concerning the: Importation Bond Structure. This request for comment is being made pursuant to the Paperwork...: Title: Importation Bond Structure. OMB Number: 1651-0050. Form Numbers: 301 and 5297. Abstract:...

  15. Photon Production through Multi-step Processes Important in Nuclear Fluorescence Experiments

    SciTech Connect

    Hagmann, C; Pruet, J

    2006-10-26

    The authors present calculations describing the production of photons through multi-step processes occurring when a beam of gamma rays interacts with a macroscopic material. These processes involve the creation of energetic electrons through Compton scattering, photo-absorption and pair production, the subsequent scattering of these electrons, and the creation of energetic photons occurring as these electrons are slowed through Bremsstrahlung emission. Unlike single Compton collisions, during which an energetic photon that is scattered through a large angle loses most of its energy, these multi-step processes result in a sizable flux of energetic photons traveling at large angles relative to an incident photon beam. These multi-step processes are also a key background in experiments that measure nuclear resonance fluorescence by shining photons on a thin foil and observing the spectrum of back-scattered photons. Effective cross sections describing the production of backscattered photons are presented in a tabular form that allows simple estimates of backgrounds expected in a variety of experiments. Incident photons with energies between 0.5 MeV and 8 MeV are considered. These calculations of effective cross sections may be useful for those designing NRF experiments or systems that detect specific isotopes in well-shielded environments through observation of resonance fluorescence.

  16. Importin 8 mediates m7G cap-sensitive nuclear import of the eukaryotic translation initiation factor eIF4E

    PubMed Central

    Volpon, Laurent; Culjkovic-Kraljacic, Biljana; Osborne, Michael J.; Ramteke, Anup; Sun, Qingxiang; Niesman, Ashley; Chook, Yuh Min; Borden, Katherine L. B.

    2016-01-01

    Regulation of nuclear-cytoplasmic trafficking of oncoproteins is critical for growth homeostasis. Dysregulated trafficking contributes to malignancy, whereas understanding the process can reveal unique therapeutic opportunities. Here, we focus on eukaryotic translation initiation factor 4E (eIF4E), a prooncogenic protein highly elevated in many cancers, including acute myeloid leukemia (AML). Typically, eIF4E is localized to both the nucleus and cytoplasm, where it acts in export and translation of specific methyl 7-guanosine (m7G)–capped mRNAs, respectively. Nuclear accumulation of eIF4E in patients who have AML is correlated with increased eIF4E-dependent export of transcripts encoding oncoproteins. The subcellular localization of eIF4E closely correlates with patients’ responses. During clinical responses to the m7G-cap competitor ribavirin, eIF4E is mainly cytoplasmic. At relapse, eIF4E reaccumulates in the nucleus, leading to elevated eIF4E-dependent mRNA export. We have identified importin 8 as a factor that directly imports eIF4E into the nucleus. We found that importin 8 is highly elevated in untreated patients with AML, leading to eIF4E nuclear accumulation. Importin 8 only imports cap-free eIF4E. Cap-dependent changes to the structure of eIF4E underpin this selectivity. Indeed, m7G cap analogs or ribavirin prevents nuclear entry of eIF4E, which mirrors the trafficking phenotypes observed in patients with AML. Our studies also suggest that nuclear entry is important for the prooncogenic activity of eIF4E, at least in this context. These findings position nuclear trafficking of eIF4E as a critical step in its regulation and position the importin 8–eIF4E complex as a novel therapeutic target. PMID:27114554

  17. Iso standardization of theoretical activity evaluation method for low and intermediate level activated waste generated at nuclear power plants

    SciTech Connect

    Makoto Kashiwagi; Garamszeghy, Mike; Lantes, Bertrand; Bonne, Sebastien; Pillette-Cousin, Lucien; Leganes, Jose Luis; Volmert, Ben; James, David W.

    2013-07-01

    Disposal of low-and intermediate-level activated waste generated at nuclear power plants is being planned or carried out in many countries. The radioactivity concentrations and/or total quantities of long-lived, difficult-to-measure nuclides (DTM nuclides), such as C-14, Ni-63, Nb-94, α emitting nuclides etc., are often restricted by the safety case for a final repository as determined by each country's safety regulations, and these concentrations or amounts are required to be known and declared. With respect to waste contaminated by contact with process water, the Scaling Factor method (SF method), which is empirically based on sampling and analysis data, has been applied as an important method for determining concentrations of DTM nuclides. This method was standardized by the International Organization for Standardization (ISO) and published in 2007 as ISO21238 'Scaling factor method to determine the radioactivity of low and intermediate-level radioactive waste packages generated at nuclear power plants' [1]. However, for activated metal waste with comparatively high concentrations of radioactivity, such as may be found in reactor control rods and internal structures, direct sampling and radiochemical analysis methods to evaluate the DTM nuclides are limited by access to the material and potentially high personnel radiation exposure. In this case, theoretical calculation methods in combination with empirical methods based on remote radiation surveys need to be used to best advantage for determining the disposal inventory of DTM nuclides while minimizing exposure to radiation workers. Pursuant to this objective a standard for the theoretical evaluation of the radioactivity concentration of DTM nuclides in activated waste, is in process through ISO TC85/SC5 (ISO Technical Committee 85: Nuclear energy, nuclear technologies, and radiological protection; Subcommittee 5: Nuclear fuel cycle). The project team for this ISO standard was formed in 2011 and is composed of

  18. Physical activity: health outcomes and importance for public health policy.

    PubMed

    Haskell, William L; Blair, Steven N; Hill, James O

    2009-10-01

    This manuscript presents a brief summary of the substantial data supporting an inverse relationship between the amount of habitual physical activity performed and a variety of negative health outcomes throughout the lifespan. It points out that despite these data a large segment of the US population remain insufficiently active resulting in a high population attributable risk for chronic disease due to inactivity. The accumulated data support the need for more comprehensive health promoting physical activity policies and programs, especially for the economically and socially disadvantaged and medically underserved.

  19. Importance of thermal nonequilibrium considerations for the simulation of nuclear reactor LOCA transients. [PWR

    SciTech Connect

    Fischer, S.R.; Nelson, R.A.; Sullivan, L.H.

    1980-01-01

    The purpose of this paper is to show the importance of considering thermal nonequilibrium effects in computer simulations of the refill and reflood portions of pressurized water reactor (PWR) loss-of-coolnat accident (LOCA) transients. Although RELAP4 assumes thermodynamic equilibrium between phases, models that account for the nonequilibrium phenomena associated with the mixing of subcooled emergency cooling water with steam and the superheating of vapor in the presence of liquid droplets have recently been incorporated into the code. Code calculated results, both with and without these new models, have been compared with experimental test data to assess the importance of including thermal nonequilibrium phenomena in computer code simulations.

  20. [Importance of rapid gradient echo sequences for nuclear magnetic tomographic diagnosis of joint diseases].

    PubMed

    König, H; Deimling, M

    1986-09-01

    The possibilities and limitations of rapid echo acquisition methods in various posttraumatic, degenerative and inflammatory diseases of the joints are demonstrated and discussed on the basis of a pilot study. It is evident that specific information important for clarifying effusions and meniscopathies is supplied especially by "water"-like images. Limitations of spatial resolution are at present still a limiting factor.

  1. Shear stress activation of nuclear receptor PXR in endothelial detoxification.

    PubMed

    Wang, Xiaohong; Fang, Xi; Zhou, Jing; Chen, Zhen; Zhao, Beilei; Xiao, Lei; Liu, Ao; Li, Yi-Shuan J; Shyy, John Y-J; Guan, Youfei; Chien, Shu; Wang, Nanping

    2013-08-06

    Endothelial cells (ECs) are constantly exposed to xenobiotics and endobiotics or their metabolites, which perturb EC function, as well as to shear stress, which plays a crucial role in vascular homeostasis. Pregnane X receptor (PXR) is a nuclear receptor and a key regulator of the detoxification of xeno- and endobiotics. Here we show that laminar shear stress (LSS), the atheroprotective flow, activates PXR in ECs, whereas oscillatory shear stress, the atheroprone flow, suppresses PXR. LSS activation of PXR in cultured ECs led to the increased expression of a PXR target gene, multidrug resistance 1 (MDR1). An in vivo study using rats showed that the expression of MDR1 was significantly higher in the endothelium from the descending thoracic aorta, where flow is mostly laminar, than from the inner curvature of aortic arch, where flow is disturbed. Functionally, LSS-activated PXR protects ECs from apoptosis triggered by doxorubicin via the induction of MDR1 and other detoxification genes. PXR also suppressed the expression of proinflammatory adhesion molecules and monocyte adhesion in response to TNF-α and lipopolysaccharide. Overexpression of a constitutively active PXR in rat carotid arteries potently attenuated proinflammatory responses. In addition, cDNA microarray revealed a large number of the PXR-activated endothelial genes whose products are responsible for major steps of detoxification, including phase I and II metabolizing enzymes and transporters. These detoxification genes in ECs are induced by LSS in ECs in a PXR-dependent manner. In conclusion, our results indicate that PXR represents a flow-activated detoxification system to protect ECs against damage by xeno- and endobiotics.

  2. Shear stress activation of nuclear receptor PXR in endothelial detoxification

    PubMed Central

    Wang, Xiaohong; Fang, Xi; Zhou, Jing; Chen, Zhen; Zhao, Beilei; Xiao, Lei; Liu, Ao; Li, Yi-Shuan J.; Shyy, John Y.-J.; Guan, Youfei; Chien, Shu; Wang, Nanping

    2013-01-01

    Endothelial cells (ECs) are constantly exposed to xenobiotics and endobiotics or their metabolites, which perturb EC function, as well as to shear stress, which plays a crucial role in vascular homeostasis. Pregnane X receptor (PXR) is a nuclear receptor and a key regulator of the detoxification of xeno- and endobiotics. Here we show that laminar shear stress (LSS), the atheroprotective flow, activates PXR in ECs, whereas oscillatory shear stress, the atheroprone flow, suppresses PXR. LSS activation of PXR in cultured ECs led to the increased expression of a PXR target gene, multidrug resistance 1 (MDR1). An in vivo study using rats showed that the expression of MDR1 was significantly higher in the endothelium from the descending thoracic aorta, where flow is mostly laminar, than from the inner curvature of aortic arch, where flow is disturbed. Functionally, LSS-activated PXR protects ECs from apoptosis triggered by doxorubicin via the induction of MDR1 and other detoxification genes. PXR also suppressed the expression of proinflammatory adhesion molecules and monocyte adhesion in response to TNF-α and lipopolysaccharide. Overexpression of a constitutively active PXR in rat carotid arteries potently attenuated proinflammatory responses. In addition, cDNA microarray revealed a large number of the PXR-activated endothelial genes whose products are responsible for major steps of detoxification, including phase I and II metabolizing enzymes and transporters. These detoxification genes in ECs are induced by LSS in ECs in a PXR-dependent manner. In conclusion, our results indicate that PXR represents a flow-activated detoxification system to protect ECs against damage by xeno- and endobiotics. PMID:23878263

  3. Homocysteine-induced apoptosis in endothelial cells coincides with nuclear NOX2 and peri-nuclear NOX4 activity.

    PubMed

    Sipkens, Jessica A; Hahn, Nynke; van den Brand, Carlien S; Meischl, Christof; Cillessen, Saskia A G M; Smith, Desirée E C; Juffermans, Lynda J M; Musters, René J P; Roos, Dirk; Jakobs, Cornelis; Blom, Henk J; Smulders, Yvo M; Krijnen, Paul A J; Stehouwer, Coen D A; Rauwerda, Jan A; van Hinsbergh, Victor W M; Niessen, Hans W M

    2013-11-01

    Apoptosis of endothelial cells related to homocysteine (Hcy) has been reported in several studies. In this study, we evaluated whether reactive oxygen species (ROS)-producing signaling pathways contribute to Hcy-induced apoptosis induction, with specific emphasis on NADPH oxidases. Human umbilical vein endothelial cells were incubated with 0.01-2.5 mM Hcy. We determined the effect of Hcy on caspase-3 activity, annexin V positivity, intracellular NOX1, NOX2, NOX4, and p47(phox) expression and localization, nuclear nitrotyrosine accumulation, and mitochondrial membrane potential (ΔΨ m). Hcy induced caspase-3 activity and apoptosis; this effect was concentration dependent and maximal after 6-h exposure to 2.5 mM Hcy. It was accompanied by a significant increase in ΔΨ m. Cysteine was inactive on these parameters excluding a reactive thiol group effect. Hcy induced an increase in cellular NOX2, p47(phox), and NOX4, but not that of NOX1. 3D digital imaging microscopy followed by image deconvolution analysis showed nuclear accumulation of NOX2 and p47(phox) in endothelial cells exposed to Hcy, but not in control cells, which coincided with accumulation of nuclear nitrotyrosine residues. Furthermore, Hcy enhanced peri-nuclear localization of NOX4 coinciding with accumulation of peri-nuclear nitrotyrosine residues, a reflection of local ROS production. p47(phox) was also increased in the peri-nuclear region. The Hcy-induced increase in caspase-3 activity was prevented by DPI and apocynin, suggesting involvement of NOX activity. The data presented in this article reveal accumulation of nuclear NOX2 and peri-nuclear NOX4 accumulation as potential source of ROS production in Hcy-induced apoptosis in endothelial cells.

  4. Influenza A viruses escape from MxA restriction at the expense of efficient nuclear vRNP import

    PubMed Central

    Götz, Veronika; Magar, Linda; Dornfeld, Dominik; Giese, Sebastian; Pohlmann, Anne; Höper, Dirk; Kong, Byung-Whi; Jans, David A.; Beer, Martin; Haller, Otto; Schwemmle, Martin

    2016-01-01

    To establish a new lineage in the human population, avian influenza A viruses (AIV) must overcome the intracellular restriction factor MxA. Partial escape from MxA restriction can be achieved when the viral nucleoprotein (NP) acquires the critical human-adaptive amino acid residues 100I/V, 283P, and 313Y. Here, we show that introduction of these three residues into the NP of an avian H5N1 virus renders it genetically unstable, resulting in viruses harboring additional single mutations, including G16D. These substitutions restored genetic stability yet again yielded viruses with varying degrees of attenuation in mammalian and avian cells. Additionally, most of the mutant viruses lost the capacity to escape MxA restriction, with the exception of the G16D virus. We show that MxA escape is linked to attenuation by demonstrating that the three substitutions promoting MxA escape disturbed intracellular trafficking of incoming viral ribonucleoprotein complexes (vRNPs), thereby resulting in impaired nuclear import, and that the additional acquired mutations only partially compensate for this import block. We conclude that for adaptation to the human host, AIV must not only overcome MxA restriction but also an associated block in nuclear vRNP import. This inherent difficulty may partially explain the frequent failure of AIV to become pandemic. PMID:26988202

  5. Influenza A viruses escape from MxA restriction at the expense of efficient nuclear vRNP import.

    PubMed

    Götz, Veronika; Magar, Linda; Dornfeld, Dominik; Giese, Sebastian; Pohlmann, Anne; Höper, Dirk; Kong, Byung-Whi; Jans, David A; Beer, Martin; Haller, Otto; Schwemmle, Martin

    2016-03-18

    To establish a new lineage in the human population, avian influenza A viruses (AIV) must overcome the intracellular restriction factor MxA. Partial escape from MxA restriction can be achieved when the viral nucleoprotein (NP) acquires the critical human-adaptive amino acid residues 100I/V, 283P, and 313Y. Here, we show that introduction of these three residues into the NP of an avian H5N1 virus renders it genetically unstable, resulting in viruses harboring additional single mutations, including G16D. These substitutions restored genetic stability yet again yielded viruses with varying degrees of attenuation in mammalian and avian cells. Additionally, most of the mutant viruses lost the capacity to escape MxA restriction, with the exception of the G16D virus. We show that MxA escape is linked to attenuation by demonstrating that the three substitutions promoting MxA escape disturbed intracellular trafficking of incoming viral ribonucleoprotein complexes (vRNPs), thereby resulting in impaired nuclear import, and that the additional acquired mutations only partially compensate for this import block. We conclude that for adaptation to the human host, AIV must not only overcome MxA restriction but also an associated block in nuclear vRNP import. This inherent difficulty may partially explain the frequent failure of AIV to become pandemic.

  6. The Activities of the European Consortium on Nuclear Data Development and Analysis for Fusion

    SciTech Connect

    Fischer, U.; Avrigeanu, M.; Avrigeanu, V.; Cabellos, O.; Kodeli, I.; Koning, A.; Konobeyev, A.Yu.; Leeb, H.; Rochman, D.; Pereslavtsev, P.; Sauvan, P.; Sublet, J.-C.; Dupont, E.; Leichtle, D.; Izquierdo, J.

    2014-06-15

    This paper presents an overview of the activities of the European Consortium on Nuclear Data Development and Analysis for Fusion. The Consortium combines available European expertise to provide services for the generation, maintenance, and validation of nuclear data evaluations and data files relevant for ITER, IFMIF and DEMO, as well as codes and software tools required for related nuclear calculations.

  7. RECENT ACTIVITIES AT THE CENTER FOR SPACE NUCLEAR RESEARCH FOR DEVELOPING NUCLEAR THERMAL ROCKETS

    SciTech Connect

    Robert C. O'Brien

    2001-09-01

    Nuclear power has been considered for space applications since the 1960s. Between 1955 and 1972 the US built and tested over twenty nuclear reactors/ rocket-engines in the Rover/NERVA programs. However, changes in environmental laws may make the redevelopment of the nuclear rocket more difficult. Recent advances in fuel fabrication and testing options indicate that a nuclear rocket with a fuel form significantly different from NERVA may be needed to ensure public support. The Center for Space Nuclear Research (CSNR) is pursuing development of tungsten based fuels for use in a NTR, for a surface power reactor, and to encapsulate radioisotope power sources. The CSNR Summer Fellows program has investigated the feasibility of several missions enabled by the NTR. The potential mission benefits of a nuclear rocket, historical achievements of the previous programs, and recent investigations into alternatives in design and materials for future systems will be discussed.

  8. 78 FR 75576 - Agency Information Collection Activities: Importation Bond Structure

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-12

    ... Structure AGENCY: U.S. Customs and Border Protection (CBP), Department of Homeland Security. ACTION: 60-day... Importation Bond Structure. This request for comment is being made pursuant to the Paperwork Reduction Act of... Structure. OMB Number: 1651-0050. Form Number: CBP Forms 301 and 5297. Abstract: Bonds are used to...

  9. Recent Nuclear Astrophysics Data Activities in the US

    SciTech Connect

    Bardayan, D.W.; Blackmon, J.C.; Browne, E.; Firestone, R.B.; Hale, G.M.; Hoffman, R.D.; Ma, Z.; McLane, V.; Norman, E.B.; Shu, N.; Smith, D.L.; Smith, M.S.; Van Wormer, L.A.; Woosley, S.E.; Wu, S.-C.

    1999-08-30

    Measurements in nuclear physics laboratories form the empirical foundation for new, realistic, sophisticated theoretical models of a wide variety of astrophysical systems. The predictive power of these models has, in many instances, a strong dependence on the input nuclear data, and more extensive and accurate nuclear data is required for these models than ever before. Progress in astrophysics can be aided by providing scientists with more usable, accurate, and significant amounts of nuclear data in a timely fashion in formats that can be easily incorporated into their models. A number of recent data compilations, evaluations, calculations, and disseminations that address nuclear astrophysics data needs will be described.

  10. Retinoic Acid-mediated Nuclear Receptor Activation and Hepatocyte Proliferation

    PubMed Central

    Bushue, Nathan; Wan, Yu-Jui Yvonne

    2016-01-01

    Due to their well-known differentiation and apoptosis-inducing abilities, retinoic acid (RA) and its analogs have strong anti-cancer efficacy in human cancers. However, in vivo RA is a liver mitogen. While speculation has persisted that RA-mediated signaling is likely involved in hepatocyte proliferation during liver regeneration, direct evidence is still required. Findings in support of this proposition include observations that a release of retinyl palmitate (the precursor of RA) occurs in liver stellate cells following liver injury. Nevertheless, the biological action of this released vitamin A is virtually unknown. More likely is that the released vitamin A is converted to RA, the biological form, and then bound to a specific receptor (retinoid x receptor; RXRα), which is most abundantly expressed in the liver. Considering the mitogenic effects of RA, the RA-activated RXRα would likely then influence hepatocyte proliferation and liver tissue repair. At present, the mechanism by which RA stimulates hepatocyte proliferation is largely unknown. This review summarizes the activation of nuclear receptors (peroxisome proliferator activated receptor-α, pregnane x receptor, constitutive androstane receptor, and farnesoid x receptor) in an RXRα dependent manner to induce hepatocyte proliferation, providing a link between RA and its proliferative role. PMID:27635169

  11. Multiple pathways in nuclear transport: the import of U2 snRNP occurs by a novel kinetic pathway

    PubMed Central

    1991-01-01

    Protein import to the nucleus is a signal-mediated process that exhibits saturation kinetics. We investigated whether signal bearing proteins compete with U2 and U6 snRNPs during import. When injected into Xenopus oocytes, saturating concentrations of P(Lys)-BSA, a protein bearing multiple nuclear localization signals from SV40 large T- antigen, reduce the rate of [125I]P(Lys)-BSA and of [125I]nucleoplasmin import, consistent with their competing for and sharing the same limiting component of the import apparatus. In contrast, saturating concentrations of P(Lys)-BSA do not reduce the rate of HeLa [32P]U2 snRNP assembly or import. The import of U6 snRNP is also competed by P(Lys)-BSA. We conclude that U2 snRNP is imported into oocyte nuclei by a kinetic pathway that is distinct from the one followed by P(Lys)-BSA, nucleoplasmin, and U6 snRNP. PMID:1824847

  12. Anti-nuclear weapons activism in the United States and Great Britain: a comparative analysis

    SciTech Connect

    Sussman, G.

    1987-01-01

    This study is a response to the lacuna in empirical research into political activism and the nuclear issue and seeks to ascertain the social and value characteristics, political attitudes, and political behavior of activists in the United States and Great Britain. Consideration is also given to gender differences in light of evidence of an emerging gender gap in these two countries. The study investigates the common forces cited in two sets of literature - post-industrialism and anti-nuclear weapons movements - which provide a framework for analysis. Survey research data is employed to assess cross-national similarities and differences. The findings obtained indicate that while American and British activists exhibit common social and value characteristics, British activists appear more integrated in their political opposition to nuclear weapons compared with their American counterparts. Survey results indicate that the political-action repertoire of these activists is quite diverse, suggesting a new style of politics in advanced industrial democracies. Gender-based analysis reveals two important findings. First, activist American men differ significantly from the other three social groups in their attitudes towards nuclear weapons. Second, activist women in both national settings participate at a level equal to or exceeding that of activist men.

  13. Nuclear localization of platelet-activating factor receptor controls retinal neovascularization

    PubMed Central

    K Bhosle, Vikrant; Rivera, José Carlos; Zhou, Tianwei (Ellen); Omri, Samy; Sanchez, Melanie; Hamel, David; Zhu, Tang; Rouget, Raphael; Rabea, Areej Al; Hou, Xin; Lahaie, Isabelle; Ribeiro-da-Silva, Alfredo; Chemtob, Sylvain

    2016-01-01

    Platelet-activating factor (PAF) is a pleiotropic phospholipid with proinflammatory, procoagulant and angiogenic actions on the vasculature. We and others have reported the presence of PAF receptor (Ptafr) at intracellular sites such as the nucleus. However, mechanisms of localization and physiologic functions of intracellular Ptafr remain poorly understood. We hereby identify the importance of C-terminal motif of the receptor and uncover novel roles of Rab11a GTPase and importin-5 in nuclear translocation of Ptafr in primary human retinal microvascular endothelial cells. Nuclear localization of Ptafr is independent of exogenous PAF stimulation as well as intracellular PAF biosynthesis. Moreover, nuclear Ptafr is responsible for the upregulation of unique set of growth factors, including vascular endothelial growth factor, in vitro and ex vivo. We further corroborate the intracrine PAF signaling, resulting in angiogenesis in vivo, using Ptafr antagonists with distinct plasma membrane permeability. Collectively, our findings show that nuclear Ptafr translocates in an agonist-independent manner, and distinctive functions of Ptafr based on its cellular localization point to another dimension needed for pharmacologic selectivity of drugs. PMID:27462464

  14. Nuclear actin activates human transcription factor genes including the OCT4 gene.

    PubMed

    Yamazaki, Shota; Yamamoto, Koji; Tokunaga, Makio; Sakata-Sogawa, Kumiko; Harata, Masahiko

    2015-01-01

    RNA microarray analyses revealed that nuclear actin activated many human transcription factor genes including OCT4, which is required for gene reprogramming. Oct4 is known to be activated by nuclear actin in Xenopus oocytes. Our findings imply that this process of OCT4 activation is conserved in vertebrates and among cell types and could be used for gene reprogramming of human cells.

  15. Identification of Gene Markers for Activation of the Nuclear Receptor Pregnane X Receptor

    EPA Science Inventory

    Many environmentally-relevant chemicals and drugs activate the nuclear receptor pregnane X receptor (PXR). Activation of PXR in the mouse liver can lead to increases in liver weight in part through increased hepatocyte replication similar to chemicals that activate other nuclear ...

  16. Compounds active against cell walls of medically important fungi.

    PubMed Central

    Hector, R F

    1993-01-01

    A number of substances that directly or indirectly affect the cell walls of fungi have been identified. Those that actively interfere with the synthesis or degradation of polysaccharide components share the property of being produced by soil microbes as secondary metabolites. Compounds specifically interfering with chitin or beta-glucan synthesis have proven effective in studies of preclinical models of mycoses, though they appear to have a restricted spectrum of coverage. Semisynthetic derivatives of some of the natural products have offered improvements in activity, toxicology, or pharmacokinetic behavior. Compounds which act on the cell wall indirectly or by a secondary mechanism of action, such as the azoles, act against diverse fungi but are usually fungistatic in nature. Overall, these compounds are attractive candidates for further development. PMID:8457977

  17. Active Neutron-Based Interrogation System with D-D Neutron Source for Detection of Special Nuclear Materials

    NASA Astrophysics Data System (ADS)

    Takahashi, Y.; Misawa, T.; Yagi, T.; Pyeon, C. H.; Kimura, M.; Masuda, K.; Ohgaki, H.

    2015-10-01

    The detection of special nuclear materials (SNM) is an important issue for nuclear security. The interrogation systems used in a sea port and an airport are developed in the world. The active neutron-based interrogation system is the one of the candidates. We are developing the active neutron-based interrogation system with a D-D fusion neutron source for the nuclear security application. The D-D neutron source is a compact discharge-type fusion neutron source called IEC (Inertial-Electrostatic Confinement fusion) device which provides 2.45 MeV neutrons. The nuclear materials emit the highenergy neutrons by fission reaction. High-energy neutrons with energies over 2.45 MeV amount to 30% of all the fission neutrons. By using the D-D neutron source, the detection of SNMs is considered to be possible with the attention of fast neutrons if there is over 2.45 MeV. Ideally, neutrons at En>2.45 MeV do not exist if there is no nuclear materials. The detection of fission neutrons over 2.45 MeV are hopeful prospect for the detection of SNM with a high S/N ratio. In the future, the experiments combined with nuclear materials and a D-D neutron source will be conducted. Furthermore, the interrogation system will be numerically investigated by using nuclear materials, a D-D neutron source, and a steel container.

  18. Measurements and analysis of alpha-induced reactions of importance for nuclear astrophysics

    NASA Astrophysics Data System (ADS)

    de Messieres, Genevieve Escande

    2011-11-01

    Reactions during stellar helium burning are of primary importance for understanding nucleosynthesis. A detailed understanding of the critical reaction chain 4He(2alpha, gamma)12C( alpha, gamma)16O(alpha, gamma) 20Ne is necessary both because it is the primary energy source and because it determines the ratio of 12C to 16O produced, which in turn significantly effects subsequent nucleosynthesis. Also during Helium burning, the reactions 22Ne(alpha, n)25Mg and 22Ne(alpha, gamma )26Mg are crucial in determining the amount of neutrons available for the astrophysical s-process. This thesis presents new experimental results concerning the 16O(alpha, gamma) 20Ne, 22Ne(alpha, n)25Mg, and 22Ne(alpha, gamma)26Mg reaction rates. These results are then applied to the calculation of the associated stellar reaction rates in order to achieve better accuracy.

  19. Nuclear Gln3 Import Is Regulated by Nitrogen Catabolite Repression Whereas Export Is Specifically Regulated by Glutamine.

    PubMed

    Rai, Rajendra; Tate, Jennifer J; Shanmuganatham, Karthik; Howe, Martha M; Nelson, David; Cooper, Terrance G

    2015-11-01

    Gln3, a transcription activator mediating nitrogen-responsive gene expression in Saccharomyces cerevisiae, is sequestered in the cytoplasm, thereby minimizing nitrogen catabolite repression (NCR)-sensitive transcription when cells are grown in nitrogen-rich environments. In the face of adverse nitrogen supplies, Gln3 relocates to the nucleus and activates transcription of the NCR-sensitive regulon whose products transport and degrade a variety of poorly used nitrogen sources, thus expanding the cell's nitrogen-acquisition capability. Rapamycin also elicits nuclear Gln3 localization, implicating Target-of-rapamycin Complex 1 (TorC1) in nitrogen-responsive Gln3 regulation. However, we long ago established that TorC1 was not the sole regulatory system through which nitrogen-responsive regulation is achieved. Here we demonstrate two different ways in which intracellular Gln3 localization is regulated. Nuclear Gln3 entry is regulated by the cell's overall nitrogen supply, i.e., by NCR, as long accepted. However, once within the nucleus, Gln3 can follow one of two courses depending on the glutamine levels themselves or a metabolite directly related to glutamine. When glutamine levels are high, e.g., glutamine or ammonia as the sole nitrogen source or addition of glutamine analogues, Gln3 can exit from the nucleus without binding to DNA. In contrast, when glutamine levels are lowered, e.g., adding additional nitrogen sources to glutamine-grown cells or providing repressive nonglutamine nitrogen sources, Gln3 export does not occur in the absence of DNA binding. We also demonstrate that Gln3 residues 64-73 are required for nuclear Gln3 export.

  20. Nuclear factor of activated T-cells (NFAT) plays a role in SV40 infection

    SciTech Connect

    Manley, Kate; O'Hara, Bethany A.; Atwood, Walter J.

    2008-03-01

    Recent evidence highlighted a role for the transcription factor, nuclear factor of activated T-cells (NFAT), in the transcription of the human polyomavirus JCV. Here we show that NFAT is also important in the transcriptional control of the related polyomavirus, Simian Virus 40 (SV40). Inhibition of NFAT activity reduced SV40 infection of Vero, 293A, and HeLa cells, and this block occurred at the stage of viral transcription. Both NFAT3 and NFAT4 bound to the SV40 promoter through {kappa}B sites located within the 72 bp repeated enhancer region. In Vero cells, NFAT was involved in late transcription, but in HeLa and 293A cells both early and late viral transcription required NFAT activity. SV40 large T-Ag was found to increase NFAT activity and provided a positive feedback loop to transactivate the SV40 promoter.

  1. Pakistan’s Nuclear Proliferation Activities and the Recommendations of the 9/11 Commission: U.S. Policy Constraints and Options

    DTIC Science & Technology

    2005-01-25

    longer critical to U.S. regional security policy.15 India and Pakistan’s May 1998 Nuclear Tests and the Decline of Sanctions as a U.S. Nonproliferation...strained relations to a critical regional ally, but they put the U.S. Government once again at a disadvantage in dealing with Pakistan’s nuclear ...activities. A number of nonproliferation experts agree with the critical importance of keeping Pakistan’s nuclear weapons and materials out of the reach of

  2. Pakistan’s Nuclear Proliferation Activities and the Recommendations of the 9/11 Commission: U.S. Policy Constraints and Options

    DTIC Science & Technology

    2005-05-24

    with the critical importance of keeping Pakistan’s nuclear weapons and materials out of the reach of terrorists, but they disagree with the Commission’s...activities and may even have used Khan and KRL as a decoy to divert attention from PAEC, where the most critical work on nuclear weapons was being...citizens of other countries, including those in the West. They also criticized the IAEA and the United States for turning a blind eye to the nuclear

  3. 10 CFR 73.55 - Requirements for physical protection of licensed activities in nuclear power reactors against...

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... activities in nuclear power reactors against radiological sabotage. 73.55 Section 73.55 Energy NUCLEAR... Requirements at Fixed Sites § 73.55 Requirements for physical protection of licensed activities in nuclear... objective to provide high assurance that activities involving special nuclear material are not inimical...

  4. 10 CFR 73.55 - Requirements for physical protection of licensed activities in nuclear power reactors against...

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... activities in nuclear power reactors against radiological sabotage. 73.55 Section 73.55 Energy NUCLEAR... Requirements at Fixed Sites § 73.55 Requirements for physical protection of licensed activities in nuclear... objective to provide high assurance that activities involving special nuclear material are not inimical...

  5. Nuclear localization of Lyn tyrosine kinase mediated by inhibition of its kinase activity

    SciTech Connect

    Ikeda, Kikuko; Nakayama, Yuji; Togashi, Yuuki; Obata, Yuuki; Kuga, Takahisa; Kasahara, Kousuke; Fukumoto, Yasunori; Yamaguchi, Naoto

    2008-11-01

    Src-family kinases, cytoplasmic enzymes that participate in various signaling events, are found at not only the plasma membrane but also subcellular compartments, such as the nucleus, the Golgi apparatus and late endosomes/lysosomes. Lyn, a member of the Src-family kinases, is known to play a role in DNA damage response and cell cycle control in the nucleus. However, it is still unclear how the localization of Lyn to the nucleus is regulated. Here, we investigated the mechanism of the distribution of Lyn between the cytoplasm and the nucleus in epitheloid HeLa cells and hematopoietic THP-1 cells. Lyn was definitely detected in purified nuclei by immunofluorescence and immunoblotting analyses. Nuclear accumulation of Lyn was enhanced upon treatment of cells with leptomycin B (LMB), an inhibitor of Crm1-mediated nuclear export. Moreover, Lyn mutants lacking the sites for lipid modification were highly accumulated in the nucleus upon LMB treatment. Intriguingly, inhibition of the kinase activity of Lyn by SU6656, Csk overexpression, or point mutation in the ATP-binding site induced an increase in nuclear Lyn levels. These results suggest that Lyn being imported into and rapidly exported from the nucleus preferentially accumulates in the nucleus by inhibition of the kinase activity and lipid modification.

  6. Nuclear Envelope Lamin-A Couples Actin Dynamics with Immunological Synapse Architecture and T Cell Activation

    PubMed Central

    González-Granado, José María; Trigueros-Motos, Laia; Cibrián, Danay; Morlino, Giulia; Blanco-Berrocal, Marta; Osorio, Fernando Garcia; Freije, José María Pérez; López-Otín, Carlos; Sánchez-Madrid, Francisco; Andrés, Vicente

    2014-01-01

    In many cell types, nuclear A-type lamins have been implicated in structural and functional activities, including higher-order genome organization, DNA replication and repair, gene transcription, and signal transduction. However, their role in specialized immune cells remains largely unexplored. Here, we showed that the abundance of A-type lamins is almost negligible in resting naïve T lymphocytes, but that it is substantially increased upon activation of the T cell receptor (TCR), and is an early event that accelerates formation of the immunological synapse between T cells and antigen-presenting cells. We found that lamin-A enhanced the polymerization of F-actin in T cells, a critical step for immunological synapse formation, by physically connecting the nucleus to the plasma membrane through the linker of nucleoskeleton and cytoskeleton (LINC) complex. We also showed that lamin-A played a key role in other membrane, cytoplasmic, and nuclear events related to TCR activation, including receptor-clustering, downstream signaling, and target gene expression. Notably, the presence of lamin-A was associated with enhanced extracellular signal–regulated kinase 1/2 signaling, and pharmacological inhibition of this pathway reduced the extent of lamin-A–dependent T cell activation. Moreover, mice deficient in lamin-A exhibited impaired T cell responses in vivo. These findings underscore the importance of A-type lamins for TCR activation, and identify lamin-A as a previously unappreciated regulator of the immune response. PMID:24757177

  7. Auxin responsiveness of the MONOPTEROS-BODENLOS module in primary root initiation critically depends on the nuclear import kinetics of the Aux/IAA inhibitor BODENLOS.

    PubMed

    Herud, Ole; Weijers, Dolf; Lau, Steffen; Jürgens, Gerd

    2016-01-01

    Primary root formation in early embryogenesis of Arabidopsis thaliana is initiated with the specification of a single cell called hypophysis. This initial step requires the auxin-dependent release of the transcription factor MONOPTEROS (MP, also known as ARF5) from its inhibition by the Aux/IAA protein BODENLOS (BDL, also known as IAA12). Auxin-insensitive bdl mutant embryos and mp loss-of-function embryos fail to specify the hypophysis, giving rise to rootless seedlings. A suppressor screen of rootless bdl mutant seedlings yielded a mutation in the nuclear import receptor IMPORTIN-ALPHA 6 (IMPα6) that promoted primary root formation through rescue of the embryonic hypophysis defects, without causing additional phenotypic changes. Aux/IAA proteins are continually synthesized and degraded, which is essential for rapid transcriptional responses to changing auxin concentrations. Nuclear translocation of bdl:3×GFP was slowed down in impα6 mutants as measured by fluorescence recovery after photobleaching (FRAP) analysis, which correlated with the reduced inhibition of MP by bdl in transient expression assays in impα6 knock-down protoplasts. The MP-BDL module acts like an auxin-triggered genetic switch because MP activates its own expression as well as the expression of its inhibitor BDL. Using an established simulation model, we determined that the reduced nuclear translocation rate of BDL in impα6 mutant embryos rendered the auxin-triggered switch unstable, impairing the fast response to changes in auxin concentration. Our results suggest that the instability of the inhibitor BDL necessitates a fast nuclear uptake in order to reach the critical threshold level required for auxin responsiveness of the MP-BDL module in primary root initiation.

  8. [Amino acids 395-416 in DNA binding domain of STAT4 is involved in IL-12-induced nuclear import of STAT4].

    PubMed

    Huang, Yu-Mei; Wen, Ya-Ping; Li, Xuan-An; Yuan, Yuan; Luo, Qi-Zhi; Li, Ming

    2012-08-25

    The purpose of the present study is to explore the mechanism of IL-12-induced nuclear import of signal transducer and activator of transcription 4 (STAT4). Assayed by analyses of homology alignment of STATs, amino acids 395-416 in DNA binding domain was found to be a potential dimer-specific nuclear localization signal (dsNLS) of STAT4. Therefore, several plasmids were constructed. Wild-type STAT4 was inserted into the SalI and BamHI sites of pEGFP-C1 for the construction of plasmid pEGFP-STAT4. The DNA fragment of STAT4 with the deletion of amino acids 395-416 was amplified by RCR and introduced into the SalI and BamHI sites of pEGFP-C1 which was named pEGFP-STAT4-Del. Classic NLS DNA sequence of SV40 T antigen was inserted into the XhoI and HindIII sites of pEGFP-C1. This plasmid was named as pEGFP-NLS and used as a positive control. Plasmid pEGFP-NLS-STAT4-Del was constructed by inserting STAT4-Del into SalI and BamHI sites of pEGFP-NLS. These plasmids were transiently transfected into Caski cells, respectively. The results showed that, after these transfected cells were stimulated by IL-12, wild type STAT4 existed in the cytoplasm at 0 min, and was predominantly localized to the nucleus at 45 min, and distributed in both cytoplasm and nucleus at 60 min, suggesting that STAT4 translocates from cytoplasm into nucleus and finally re-entries into the cytoplasm during the stimulation of IL-12. However, deletion mutant of STAT4 was arrested in cytoplasm during the IL-12 stimulation. Leptomycin B, which specifically blocks protein export from nucleus into cytoplasm, was used to further demonstrate whether STAT4-Del is transferred into nucleus even with stimulation of IL-12. After the transfected cells were pre-treated by leptomycin B, the wild type STAT4 was mainly localized in nucleus after the IL-12 stimulation, suggesting that STAT4 was translocated from cytoplasm into nucleus by the stimulation of IL-12. On the other hand, the deletion mutant of STAT4 distributed

  9. Nuclear import of cutaneous beta genus HPV8 E7 oncoprotein is mediated by hydrophobic interactions between its zinc-binding domain and FG nucleoporins

    PubMed Central

    Onder, Zeynep; Moroianu, Junona

    2013-01-01

    We have previously discovered and characterized the nuclear import pathways for the E7 oncoproteins of mucosal alpha genus HPVs, type 16 and 11. Here we investigated the nuclear import of cutaneous beta genus HPV8 E7 protein using confocal microscopy after transfections of HeLa cells with EGFP-8E7 and mutant plasmids and nuclear import assays in digitonin-permeabilized HeLa cells. We determined that HPV8 E7 contains a nuclear localization signal (NLS) within its zinc-binding domain that mediates its nuclear import. Furthermore, we discovered that a mostly hydrophobic patch 65LRLFV69 within the zinc-binding domain is essential for the nuclear import and localization of HPV8 E7 via hydrophobic interactions with the FG nucleoporins Nup62 and Nup153. Substitution of the hydrophobic residues within the 65LRLFV69 patch to alanines, and not R66A mutation, disrupt the interactions between the 8E7 zinc-binding domain and Nup62 and Nup153 and consequently inhibit nuclear import of HPV8 E7. PMID:24418548

  10. The active liquid Earth - importance of temporal and spatial variability

    NASA Astrophysics Data System (ADS)

    Arheimer, Berit

    2016-04-01

    The Planet Earth is indeed liquid and active - 71 percent of its surface is water-covered and this water never rests. Thanks to the water cycle, our planet's water supply is constantly moving from one place to another and from one form to another. Only 2.5% of the water is freshwater and it exists in the air as water vapor; it hits the ground as rain and snow; it flows on the surface from higher to lower altitudes in rivers, lakes, and glaciers; and it flows in the ground in soil, aquifers, and in all living organisms until it reaches the sea. On its way over the Earth's crust, some returns quickly to vapor again, while some is trapped and exposed to many "fill and spill" situations for a long journey. The variability in the water balance is crucial for hydrological understanding and modelling. The water cycle may appear simple, but magnitudes and rates in fluxes are very different from one place to another, resulting from variable drivers such as solar energy, precipitation and gravity in co-evolution with geology, soil, vegetation and fauna. The historical evolution, the temporal fluxes and diversity in space continue to fascinate hydrological scientists. Specific physical processes may be well known, but their boundary conditions, interactions and rate often remain unknown at a specific site and are difficult to monitor in nature. This results in mysterious features where trends in drivers do not match runoff, like the Sahelian Paradox or discharge to the Arctic Ocean. Humans have always interfered with the water cycle and engineering is fundamental for water regulation and re-allocation. Some 80% of the river flow from the northern part of the Earth is affected by fragmentation of the river channels by dams. In water management, there is always a tradeoff between upstream and downstream activities, not only regarding total water quantities but also for temporal patterns and water quality aspects. Sharing a water resource can generate conflicts but geopolitical

  11. Nuclear Thermal Rocket Element Environmental Simulator (NTREES) Upgrade Activities

    NASA Technical Reports Server (NTRS)

    Emrich, William J., Jr.

    2014-01-01

    Over the past year the Nuclear Thermal Rocket Element Environmental Simulator (NTREES) has been undergoing a significant upgrade beyond its initial configuration. The NTREES facility is designed to perform realistic non-nuclear testing of nuclear thermal rocket (NTR) fuel elements and fuel materials. Although the NTREES facility cannot mimic the neutron and gamma environment of an operating NTR, it can simulate the thermal hydraulic environment within an NTR fuel element to provide critical information on material performance and compatibility. The first phase of the upgrade activities which was completed in 2012 in part consisted of an extensive modification to the hydrogen system to permit computer controlled operations outside the building through the use of pneumatically operated variable position valves. This setup also allows the hydrogen flow rate to be increased to over 200 g/sec and reduced the operation complexity of the system. The second stage of modifications to NTREES which has just been completed expands the capabilities of the facility significantly. In particular, the previous 50 kW induction power supply has been replaced with a 1.2 MW unit which should allow more prototypical fuel element temperatures to be reached. The water cooling system was also upgraded to so as to be capable of removing 100% of the heat generated during. This new setup required that the NTREES vessel be raised onto a platform along with most of its associated gas and vent lines. In this arrangement, the induction heater and water systems are now located underneath the platform. In this new configuration, the 1.2 MW NTREES induction heater will be capable of testing fuel elements and fuel materials in flowing hydrogen at pressures up to 1000 psi at temperatures up to and beyond 3000 K and at near-prototypic reactor channel power densities. NTREES is also capable of testing potential fuel elements with a variety of propellants, including hydrogen with additives to inhibit

  12. Nuclear Thermal Rocket Element Environmental Simulator (NTREES) Upgrade Activities

    NASA Technical Reports Server (NTRS)

    Emrich, William J. Jr.; Moran, Robert P.; Pearson, J. Boise

    2012-01-01

    To support the on-going nuclear thermal propulsion effort, a state-of-the-art non nuclear experimental test setup has been constructed to evaluate the performance characteristics of candidate fuel element materials and geometries in representative environments. The facility to perform this testing is referred to as the Nuclear Thermal Rocket Element Environment Simulator (NTREES). This device can simulate the environmental conditions (minus the radiation) to which nuclear rocket fuel components will be subjected during reactor operation. Test articles mounted in the simulator are inductively heated in such a manner so as to accurately reproduce the temperatures and heat fluxes which would normally occur as a result of nuclear fission and would be exposed to flowing hydrogen. Initial testing of a somewhat prototypical fuel element has been successfully performed in NTREES and the facility has now been shutdown to allow for an extensive reconfiguration of the facility which will result in a significant upgrade in its capabilities

  13. A nuclear localization signal and the C-terminal omega sequence in the Agrobacterium tumefaciens VirD2 endonuclease are important for tumor formation.

    PubMed

    Shurvinton, C E; Hodges, L; Ream, W

    1992-12-15

    The T-DNA portion of the Agrobacterium tumefaciens tumor-inducing (Ti) plasmid integrates into plant nuclear DNA. Direct repeats define the T-DNA ends; transfer begins when the VirD2 endonuclease produces a site-specific nick in the right-hand border repeat and attaches to the 5' end of the nicked strand. Subsequent events generate linear single-stranded VirD2-bound DNA molecules that include the entire T-DNA (T-strands). VirD2 protein contains a nuclear localization signal (NLS) near the C terminus and may direct bound T-strands to plant nuclei. We constructed mutations in virD2 and showed that the NLS was important for tumorigenesis, although T-strand production occurred normally in its absence. A tobacco etch virus NLS, substituted for the VirD2 NLS, restored tumor-inducing activity. Amino acids (the omega sequence) at the C terminus of VirD2, outside the NLS and the endonuclease domain, contributed significantly to tumorigenesis, suggesting that VirD2 may serve a third important function in T-DNA transfer.

  14. Nuclear import of high risk HPV16 E7 oncoprotein is mediated by its zinc-binding domain via hydrophobic interactions with Nup62

    SciTech Connect

    Eberhard, Jeremy; Onder, Zeynep; Moroianu, Junona

    2013-11-15

    We previously discovered that nuclear import of high risk HPV16 E7 is mediated by a cNLS located within the zinc-binding domain via a pathway that is independent of karyopherins/importins (Angeline et al., 2003; Knapp et al., 2009). In this study we continued our characterization of the cNLS and nuclear import pathway of HPV16 E7. We find that an intact zinc-binding domain is essential for the cNLS function in mediating nuclear import of HPV16 E7. Mutagenesis of cysteine residues to alanine in each of the two CysXXCys motifs involved in zinc-binding changes the nuclear localization of the EGFP-16E7 and 2xEGFP-16E7 mutants. We further discover that a patch of hydrophobic residues, {sub 65}LRLCV{sub 69}, within the zinc-binding domain of HPV16 E7 mediates its nuclear import via hydrophobic interactions with the FG domain of the central channel nucleoporin Nup62. - Highlights: • An intact zinc-binding domain is essential for the nuclear localization of HPV16 E7. • Identification of a hydrophobic patch that is critical for the nuclear import of HPV16 E7. • HPV16 E7 interacts via its zinc-binding domain with the FG domain of Nup62.

  15. Research activities in nuclear astrophysics and related areas

    NASA Technical Reports Server (NTRS)

    1996-01-01

    NASA/GRO grant NAG 5-2081, at the University of Chicago, has provided support for a broad program of theoretical research in nuclear astrophysics and related areas, with regard to gamma-ray and hard X-ray emission from classical nova explosions. This research emphasized the possible detection of 22Na gamma-ray line emission from nearby novae involving ONeMg white dwarfs, the detailed examination of 26Al production in novae, and the possible detection of the predicted early gamma ray emission from novae that arises from the decay of the short lived, positron emitting isotopes of CNO elements. Studies of nova related problems have consumed an increasing fraction of the Principal Investigator's research efforts over the past decade. Current research addresses problems associated with the standard model for the outbursts of the classical novae: the occurrence of thermonuclear runaways (TNR) in the accreted hydrogen rich envelopes on white dwarfs in close binary systems (see, e.g., the reviews by Truran 1982; and Shara 1989). Research in progress and planned for the next three years has three main objectives: (1) to gain an improved understanding of the early evolution of the light curves of, particularly, the fastest novae; (2) to gain an improved understanding of the relative importance of the various possible mechanisms of envelope hydrogen depletion (e.g. winds, common envelope driven mass loss, and nuclear burning) to the long term evolution of novae in outburst; and (3) to seek to provide a somewhat more definitive statement of the role of classical novae in nucleosynthesis. Our proposed 2-D studies of convection during the early phases of the TNR and our systematic attempt to incorporate an improved treatment of radiation hydrodynamics into the hydrodynamic code utilized in our calculations, are particularly relevant to the first of these objectives. Further 2-D studies of the effects of common envelope evolution are intended to provide more realistic constraints

  16. Delayed Gamma Measurements in Different Nuclear Research Reactors Bringing Out the Importance of the Delayed Contribution in Gamma Flux Calculations

    SciTech Connect

    Fourmentel, D.; Radulovic, V.; Barbot, L.; Villard, J-F.; Zerovnik, G.; Snoj, L.; Tarchalski, M.; Pytel, K.; Malouch, F.

    2015-07-01

    Neutron and gamma flux levels are key parameters in nuclear research reactors. In Material Testing Reactors, such as the future Jules Horowitz Reactor, under construction at the French Alternative Energies and Atomic Energy Commission (CEA Cadarache, France), the expected gamma flux levels are very high (nuclear heating is of the order of 20 W/g at 100 MWth). As gamma rays deposit their energy in the reactor structures and structural materials it is important to take them into account when designing irradiation devices. There are only a few sensors which allow measurements of the nuclear heating ; a recent development at the CEA Cadarache allows measurements of the gamma flux using a miniature ionization chamber (MIC). The measured MIC response is often compared with calculation using modern Monte Carlo (MC) neutron and photon transport codes, such as TRIPOLI-4 and MCNP6. In these calculations only the production of prompt gamma rays in the reactor is usually modelled thus neglecting the delayed gamma rays. Hence calculations and measurements are usually in better accordance for the neutron flux than for the gamma flux. In this paper we study the contribution of delayed gamma rays to the total MIC signal in order to estimate the systematic error in gamma flux MC calculations. In order to experimentally determine the delayed gamma flux contributions to the MIC response, we performed gamma flux measurements with CEA developed MIC at three different research reactors: the OSIRIS reactor (MTR - 70 MWth at CEA Saclay, France), the TRIGA MARK II reactor (TRIGA - 250 kWth at the Jozef Stefan Institute, Slovenia) and the MARIA reactor (MTR - 30 MWth at the National Center for Nuclear Research, Poland). In order to experimentally assess the delayed gamma flux contribution to the total gamma flux, several reactor shut down (scram) experiments were performed specifically for the purpose of the measurements. Results show that on average about 30 % of the MIC signal is due to

  17. NUCLEAR ACTIVITY IS MORE PREVALENT IN STAR-FORMING GALAXIES

    SciTech Connect

    Rosario, D. J.; Lutz, D.; Berta, S.; Popesso, P.; Genzel, R.; Saintonge, A.; Tacconi, L.; Wuyts, S. E-mail: lutz@mpe.mpg.de E-mail: popesso@mpe.mpg.de E-mail: amelie@mpe.mpg.de E-mail: swuyts@mpe.mpg.de; and others

    2013-07-01

    We explore the question of whether low and moderate luminosity active galactic nuclei (AGNs) are preferentially found in galaxies that are undergoing a transition from active star formation (SF) to quiescence. This notion has been suggested by studies of the UV-optical colors of AGN hosts, which find them to be common among galaxies in the so-called Green Valley, a region of galaxy color space believed to be composed mostly of galaxies undergoing SF quenching. Combining the deepest current X-ray and Herschel/PACS far-infrared (FIR) observations of the two Chandra Deep Fields with redshifts, stellar masses, and rest-frame photometry derived from the extensive and uniform multi-wavelength data in these fields, we compare the rest-frame U - V color distributions and star formation rate distributions of AGNs and carefully constructed samples of inactive control galaxies. The UV-to-optical colors of AGNs are consistent with equally massive inactive galaxies at redshifts out to z {approx} 2, but we show that such colors are poor tracers of SF. While the FIR distributions of both star-forming AGNs and star-forming inactive galaxies are statistically similar, we show that AGNs are preferentially found in star-forming host galaxies, or, in other words, AGNs are less likely to be found in weakly star-forming or quenched galaxies. We postulate that, among X-ray-selected AGNs of low and moderate accretion luminosities, the supply of cold gas primarily determines the accretion rate distribution of the nuclear black holes.

  18. Caspase-like activity in programmed nuclear death during conjugation of Tetrahymena thermophila.

    PubMed

    Kobayashi, T; Endoh, H

    2003-06-01

    Apoptosis, or programmed cell death, is common in a variety of eucaryotes, from unicellular protozoa to vertebrates. The ciliated protozoan Tetrahymena thermophila has a unique apoptosis-like nuclear death during conjugation, called programmed nuclear death. This death program involves nuclear condensation (pyknosis) and oligonucleosomal DNA fragmentation in the parental macronucleus. Subsequently, the condensed nucleus is entirely resorbed in the autophagosome. Here we demonstrate that caspase-8- and -9-like activity was detected, but no caspase-3-like activity, by in vitro assay during the nuclear resorption process, suggesting that caspase-like activity is associated with both programmed cell death and apoptosis-like nuclear death in Tetrahymena. The use of indicator dye to detect the loss of mitochondrial membrane potential suggested the uptake of mitochondria and the degenerating macronucleus by the autophagosome. An involvement of mitochondria in the programmed nuclear death is discussed.

  19. Active Interrogation using Photofission Technique for Nuclear Materials Control and Accountability

    SciTech Connect

    Yang, Haori

    2016-03-31

    Innovative systems with increased sensitivity and resolution are in great demand to detect diversion and to prevent misuse in support of nuclear materials management for the U.S. fuel cycle. Nuclear fission is the most important multiplicative process involved in non-destructive active interrogation. This process produces the most easily recognizable signature for nuclear materials. In addition to thermal or high-energy neutrons, high-energy gamma rays can also excite a nucleus and cause fission through a process known as photofission. Electron linear accelerators (linacs) are widely used as the interrogating photon sources for inspection methods involving photofission technique. After photofission reactions, prompt signals are much stronger than the delayed signals, but it is difficult to quantify them in practical measurements. Delayed signals are easily distinguishable from the interrogating radiation. Linac-based, advanced inspection techniques utilizing the delayed signals after photofission have been extensively studied for homeland security applications. Previous research also showed that a unique delayed gamma ray energy spectrum exists for each fissionable isotope. In this work, high-energy delayed γ-rays were demonstrated to be signatures for detection, identification, and quantification of special nuclear materials. Such γ-rays were measured in between linac pulses using independent data acquisition systems. A list-mode system was developed to measure low-energy delayed γ-rays after irradiation. Photofission product yields of 238U and 239Pu were determined based on the measured delayed γ-ray spectra. The differential yields of delayed γ-rays were also proven to be able to discriminate nuclear from non-nuclear materials. The measurement outcomes were compared with Monte Carlo simulation results. It was demonstrated that the current available codes have capabilities and limitations in the simulation of photofission process. A two

  20. Finerenone Impedes Aldosterone-dependent Nuclear Import of the Mineralocorticoid Receptor and Prevents Genomic Recruitment of Steroid Receptor Coactivator-1*

    PubMed Central

    Amazit, Larbi; Le Billan, Florian; Kolkhof, Peter; Lamribet, Khadija; Viengchareun, Say; Fay, Michel R.; Khan, Junaid A.; Hillisch, Alexander; Lombès, Marc; Rafestin-Oblin, Marie-Edith; Fagart, Jérôme

    2015-01-01

    Aldosterone regulates sodium homeostasis by activating the mineralocorticoid receptor (MR), a member of the nuclear receptor superfamily. Hyperaldosteronism leads todeleterious effects on the kidney, blood vessels, and heart. Although steroidal antagonists such as spironolactone and eplerenone are clinically useful for the treatment of cardiovascular diseases, they are associated with several side effects. Finerenone, a novel nonsteroidal MR antagonist, is presently being evaluated in two clinical phase IIb trials. Here, we characterized the molecular mechanisms of action of finerenone and spironolactone at several key steps of the MR signaling pathway. Molecular modeling and mutagenesis approaches allowed identification of Ser-810 and Ala-773 as key residues for the high MR selectivity of finerenone. Moreover, we showed that, in contrast to spironolactone, which activates the S810L mutant MR responsible for a severe form of early onset hypertension, finerenone displays strict antagonistic properties. Aldosterone-dependent phosphorylation and degradation of MR are inhibited by both finerenone and spironolactone. However, automated quantification of MR subcellular distribution demonstrated that finerenone delays aldosterone-induced nuclear accumulation of MR more efficiently than spironolactone. Finally, chromatin immunoprecipitation assays revealed that, as opposed to spironolactone, finerenone inhibits MR, steroid receptor coactivator-1, and RNA polymerase II binding at the regulatory sequence of the SCNN1A gene and also remarkably reduces basal MR and steroid receptor coactivator-1 recruitment, unraveling a specific and unrecognized inactivating mechanism on MR signaling. Overall, our data demonstrate that the highly potent and selective MR antagonist finerenone specifically impairs several critical steps of the MR signaling pathway and therefore represents a promising new generation MR antagonist. PMID:26203193

  1. Identification of a small molecule inhibitor of importin beta mediated nuclear import by confocal on-bead screening of tagged one-bead one-compound libraries

    PubMed Central

    Hintersteiner, Martin; Ambrus, Géza; Bednenko, Janna; Schmied, Mario; Knox, Andrew J.S.; Gstach, Hubert; Seifert, Jan-Marcus; Singer, Eric L.; Gerace, Larry; Auer, Manfred

    2010-01-01

    In eukaryotic cells, proteins and RNA are transported between the nucleus and the cytoplasm by nuclear import and export receptors. Over the past decade, small molecules that inhibit the nuclear export receptor CRM1 have been identified, most notably leptomycin B. However, up to now no small molecule inhibitors of nuclear import have been described. Here we have used our automated Confocal Nanoscanning and bead picking method (CONA) for on-bead screening of a one bead/one compound library to identify the first such import inhibitor, karyostatin 1A. Karyostatin 1A binds importin β with high nanomolar affinity and specifically inhibits importin α/β mediated nuclear import at low micromolar concentrations in vitro and in living cells, without perturbing transportin mediated nuclear import or CRM1 mediated nuclear export. Surface plasmon resonance binding experiments suggest that karyostatin 1A acts by disrupting the interaction between importin β and the GTPase Ran. As a selective inhibitor of the importin α/β import pathway, karyostatin 1A will provide a valuable tool for future studies of nucleocytoplasmic trafficking. PMID:20677820

  2. Determination of Importance Evaluation for Exploratory Studies Facility (ESF) Subsurface Testing Activities

    SciTech Connect

    C.J. Byrne

    2001-02-20

    This Determination of Importance Evaluation (DIE) applies to the Subsurface Exploratory Studies Facility (ESF), encompassing the Topopah Spring (TS) Loop from Station 0+00 meters (m) at the North Portal to breakthrough at the South Portal (approximately 78+77 m), and ancillary test and operation support areas including the Enhanced Characterization of the Repository Block (ECRB) Cross Drift. This evaluation applies specifically to site characterization testing activities ongoing and planned in the Subsurface ESF. ESF site characterization activities are being performed to obtain the information necessary to determine whether the Yucca Mountain Site is suitable as a geologic repository for spent nuclear fuel and high-level radioactive waste. A more detailed description of these testing activities is provided in Section 6 of this DIE. Generally, the construction and operation of excavations associated with these testing activities are evaluated in the DIE for the Subsurface ESF (CRWMS M&O 1999a) and the DIE for the ESF ECRB Cross Drift (CRWMS M&O 2000a). The scope of this DIE also entails the proposed Unsaturated Zone (UZ) Transport Test at Busted Butte. Although, not a part of the TS Loop or ECRB Cross Drift, the associated testing activities are Subsurface testing activities. Busted Butte is located to the south south-east of the TS Loop and is outside the Conceptual Controlled Area Boundary (CCAB). These activities provide access to the Calico Hills (CH) geologic structure. In the case of Busted Butte, construction and operation of excavations are evaluated herein (since this activity was not previously evaluated in CRWMS M&O 1999a). The objectives of this DIE are to determine whether Subsurface ESF testing, and associated activities, could potentially impact site characterization testing and/or the waste isolation capabilities of the site. Controls needed to limit any potential impacts are identified in Section 13. The validity and veracity of the individual

  3. Phenosafranin inhibits nuclear localization of transglutaminase 2 without affecting its transamidase activity.

    PubMed

    Furutani, Yutaka; Toguchi, Mariko; Shrestha, Rajan; Kojima, Soichi

    2017-03-01

    Transglutaminase 2 (TG2) localizes to the nucleus and induces apoptosis through a crosslinking inactivation of Sp1 in JHH-7 cells treated with acyclic retinoid. We screened an inhibitor suppressing transamidase activity in the nucleus without affecting transamidase activity itself. Phenosafranin was found to inhibit nuclear localization of EGFP-tagged TG2 and dose-dependently reduce nuclear transamidase activity without affecting the activity in a tube. We concluded that phenosafranin was a novel TG2 inhibitor capable of suppressing its nuclear localization.

  4. Decreased activity and enhanced nuclear export of CCAAT-enhancer-binding protein beta during inhibition of adipogenesis by ceramide.

    PubMed Central

    Sprott, Kam M; Chumley, Michael J; Hanson, Janean M; Dobrowsky, Rick T

    2002-01-01

    To identify novel molecular mechanisms by which ceramide regulates cell differentiation, we examined its effect on adipogenesis of 3T3-L1 preadipocytes. Hormonal stimulation of 3T3-L1 preadipocytes induced formation of triacylglycerol-laden adipocytes over 7 days; in part, via the co-ordinated action of CCAAT-enhancer-binding proteins alpha, beta and delta (C/EBP-alpha, -beta and -delta) and peroxisome-proliferator-activated receptor gamma (PPARgamma). The addition of exogenous N-acetylsphingosine (C2-ceramide) or increasing endogenous ceramide levels inhibited the expression of C/EBPalpha and PPARgamma, and blocked adipocyte development. C2-ceramide did not decrease the cellular expression of C/EBPbeta, which is required for expression of C/EBPalpha and PPARgamma, but significantly blocked its transcriptional activity from a promoter construct after 24 h. The ceramide-induced decrease in the transcriptional activity of C/EBPbeta correlated with a strong decrease in its phosphorylation, DNA-binding ability and nuclear localization at 24 h. However, ceramide did not change the nuclear level of C/EBPbeta after a period of 4 or 16 h, suggesting that it was not affecting nuclear import. CRM1 (more recently named 'exportin-1') is a nuclear membrane protein that regulates protein export from the nucleus by binding to a specific nuclear export sequence. Leptomycin B is an inhibitor of CRM1/exportin-1, and reversed the ceramide-induced decrease in nuclear C/EBPbeta at 24 h. Taken together, these data support the hypothesis that ceramide may inhibit adipogenesis, at least in part, by enhancing dephosphorylation and premature nuclear export of C/EBPbeta at a time when its maximal transcriptional activity is required to drive adipogenesis. PMID:12071851

  5. 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

  6. Determination of Importance Evaluation for Exploratory Studies Facility (ESF) Subsurface Testing Activities

    SciTech Connect

    S. Goodin

    2002-07-22

    This Determination of Importance Evaluation (DIE) applies to the Subsurface Exploratory Studies Facility (ESF), encompassing the Topopah Spring (TS) Loop from Station 0+00 meters (m) at the North Portal to breakthrough at the South Portal (approximately 78+77 m), and ancillary test and operation support areas including the Enhanced Characterization of the Repository Block (ECRB) Cross Drift. This evaluation applies specifically to site characterization testing activities ongoing and planned in the Subsurface ESF. ESF site characterization activities are being performed to obtain the information necessary to determine whether the Yucca Mountain Site is suitable as a geologic repository for spent nuclear fuel and high-level radioactive waste. A more detailed description of these testing activities is provided in Section 6 of this DIE. Generally, the construction and operation of excavations associated with these testing activities are evaluated in the DIE for the Subsurface ESF (CRWMS M&O 1999a) and the DIE for the ESF ECRB Cross Drift (CRWMS M&O 2000a). The scope of this DIE also entails the proposed Unsaturated Zone (UZ) Transport Test at Busted Butte. Although, not a part of the TS Loop or ECRB Cross Drift, the associated testing activities are Subsurface testing activities. Busted Butte is located to the south south-east of the TS Loop and is outside the Conceptual Controlled Area Boundary (CCAB). These activities provide access to the Calico Hills (CH) geologic structure. In the case of Busted Butte, construction and operation of excavations are evaluated herein (since this activity was not previously evaluated in CRWMS M&O 1999a). The objectives of this DIE are to determine whether Subsurface ESF testing, and associated activities, could potentially impact site characterization testing and/or the waste isolation capabilities of the site. Controls needed to limit any potential impacts are identified in Section 13. The validity and veracity of the individual

  7. Structural and calorimetric studies demonstrate that the hepatocyte nuclear factor 1β (HNF1β) transcription factor is imported into the nucleus via a monopartite NLS sequence.

    PubMed

    Wiedmann, Mareike M; Aibara, Shintaro; Spring, David R; Stewart, Murray; Brenton, James D

    2016-09-01

    The transcription factor hepatocyte nuclear factor 1β (HNF1β) is ubiquitously overexpressed in ovarian clear cell carcinoma (CCC) and is a potential therapeutic target. To explore potential approaches that block HNF1β transcription we have identified and characterised extensively the nuclear localisation signal (NLS) for HNF1β and its interactions with the nuclear protein import receptor, Importin-α. Pull-down assays demonstrated that the DNA binding domain of HNF1β interacted with a spectrum of Importin-α isoforms and deletion constructs tagged with eGFP confirmed that the HNF1β (229)KKMRRNR(235) sequence was essential for nuclear localisation. We further characterised the interaction between the NLS and Importin-α using complementary biophysical techniques and have determined the 2.4Å resolution crystal structure of the HNF1β NLS peptide bound to Importin-α. The functional, biochemical, and structural characterisation of the nuclear localisation signal present on HNF1β and its interaction with the nuclear import protein Importin-α provide the basis for the development of compounds targeting transcription factor HNF1β via its nuclear import pathway.

  8. Activation of nuclear factor-κB in human prostate carcinogenesis and association to biochemical relapse

    PubMed Central

    Domingo-Domenech, J; Mellado, B; Ferrer, B; Truan, D; Codony-Servat, J; Sauleda, S; Alcover, J; Campo, E; Gascon, P; Rovira, A; Ross, J S; Fernández, P L; Albanell, J

    2005-01-01

    Nuclear factor (NF)-κB/p65 regulates the transcription of a wide variety of genes involved in cell survival, invasion and metastasis. We characterised by immunohistochemistry the expression of NF-κB/p65 protein in six histologically normal prostate, 13 high-grade prostatic intraepithelial neoplasia (PIN) and 86 prostate adenocarcinoma specimens. Nuclear localisation of p65 was used as a measure of NF-κB active state. Nuclear localisation of NF-κB was only seen in scattered basal cells in normal prostate glands. Prostatic intraepithelial neoplasias exhibited diffuse and strong cytoplasmic staining but no nuclear staining. In prostate adenocarcinomas, cytoplasmic NF-κB was detected in 57 (66.3%) specimens, and nuclear NF-κB (activated) in 47 (54.7%). Nuclear and cytoplasmic NF-κB staining was not correlated (P=0.19). By univariate analysis, nuclear localisation of NF-κB was associated with biochemical relapse (P=0.0009; log-rank test) while cytoplasmic expression did not. On multivariate analysis, serum preoperative prostate specific antigen (P=0.02), Gleason score (P=0.03) and nuclear NF-κB (P=0.002) were independent predictors of biochemical relapse. These results provide novel evidence for NF-κB/p65 nuclear translocation in the transition from PIN to prostate cancer. Our findings also indicate that nuclear localisation of NF-κB is an independent prognostic factor of biochemical relapse in prostate cancer. PMID:16278667

  9. Controlling nuclear JAKs and STATs for specific gene activation by IFN{gamma}

    SciTech Connect

    Noon-Song, Ezra N.; Ahmed, Chulbul M.; Dabelic, Rea; Canton, Johnathan; Johnson, Howard M.

    2011-07-08

    Highlights: {yields} Gamma interferon (IFN{gamma}) and its receptor subunit, IFNGR1, interact with the promoter region of IFN{gamma}-associated genes along with transcription factor STAT1{alpha}. {yields} We show that activated Janus kinases pJAK2 and pJAK1 also associate with IFNGR1 in the nucleus. {yields} The activated Janus kinases are responsible for phosphorylation of tyrosine 41 on histone H3, an important epigenetic event for specific gene activation. -- Abstract: We previously showed that gamma interferon (IFN{gamma}) and its receptor subunit, IFNGR1, interacted with the promoter region of IFN{gamma}-activated genes along with transcription factor STAT1{alpha}. Recent studies have suggested that activated Janus kinases pJAK2 and pJAK1 also played a role in gene activation by phosphorylation of histone H3 on tyrosine 41. This study addresses the question of the role of activated JAKs in specific gene activation by IFN{gamma}. We carried out chromatin immunoprecipitation (ChIP) followed by PCR in IFN{gamma} treated WISH cells and showed association of pJAK1, pJAK2, IFNGR1, and STAT1 on the same DNA sequence of the IRF-1 gene promoter. The {beta}-actin gene, which is not activated by IFN{gamma}, did not show this association. The movement of activated JAK to the nucleus and the IRF-1 promoter was confirmed by the combination of nuclear fractionation, confocal microscopy and DNA precipitation analysis using the biotinylated GAS promoter. Activated JAKs in the nucleus was associated with phosphorylated tyrosine 41 on histone H3 in the region of the GAS promoter. Unphosphorylated JAK2 was found to be constitutively present in the nucleus and was capable of undergoing activation in IFN{gamma} treated cells, most likely via nuclear IFNGR1. Association of pJAK2 and IFNGR1 with histone H3 in IFN{gamma} treated cells was demonstrated by histone H3 immunoprecipitation. Unphosphorylated STAT1 protein was associated with histone H3 of untreated cells. IFN

  10. Zinc inhibits nuclear factor-kappa B activation and sensitizes prostate cancer cells to cytotoxic agents.

    PubMed

    Uzzo, Robert G; Leavis, Paul; Hatch, William; Gabai, Vladimir L; Dulin, Nickolai; Zvartau, Nadezhda; Kolenko, Vladimir M

    2002-11-01

    Prostate carcinogenesis involves transformation of zinc-accumulating normal epithelial cells to malignant cells, which do not accumulate zinc. In this study, we demonstrate by immunoblotting and immunohistochemistry that physiological levels of zinc inhibit activation of nuclear factor (NF)-kappa B transcription factor in PC-3 and DU-145 human prostate cancer cells, reduce expression of NF-kappa B-controlled antiapoptotic protein c-IAP2, and activate c-Jun NH(2)-terminal kinases. Preincubation of PC-3 cells with physiological concentrations of zinc sensitized tumor cells to tumor necrosis factor (TNF)-alpha, and paclitaxel mediated cell death as defined by terminal deoxynucleotidyl transferase-mediated nick end labeling assay. These results suggest one possible mechanism for the inhibitory effect of zinc on the development and progression of prostate malignancy and might have important consequences for the prevention and treatment of prostate cancer.

  11. Hypoxic preconditioning decreases nuclear factor κB activity via Disrupted in Schizophrenia-1.

    PubMed

    Liu, Jia-Ren; Liu, Qian; Khoury, Joseph; Li, Yue-Jin; Han, Xiao-Hui; Li, Jing; Ibla, Juan C

    2016-01-01

    Nuclear factor κB is a key mediator of inflammation during conditions of hypoxia. Here, we used models of hypoxic pre-conditioning as mechanism to decrease nuclear factor κB activity induced by hypoxia. Our initial studies suggested that Disrupted in Schizophrenia-1 may be induced by hypoxic pre-conditioning and possibly involved in the regulation of nuclear factor κB. In this study we used Disrupted in Schizophrenia-1 exogenous over-expression and knock-down to determine its effect on ataxia telangiectasia mutated--nuclear factor κB activation cascade. Our results demonstrated that hypoxic pre-conditioning significantly increased the expression of Disrupted in Schizophrenia-1 at mRNA and protein levels both in vitro and in vivo. Over-expression of Disrupted in Schizophrenia-1 significantly attenuated the hypoxia-mediated ataxia telangiectasia mutated phosphorylation and prevented its cytoplasm translocation where it functions to activate nuclear factor κB. We further determined that Disrupted in Schizophrenia-1 activated the protein phosphatase 2A, preventing the phosphorylation of ataxia telangiectasia mutated serine-1981, the main regulatory site of ataxia telangiectasia mutated activity. Cellular levels of Disrupted in Schizophrenia-1 protein significantly decreased nuclear factor κB activation profiles and pro-inflammatory gene expression. Taken together, these results demonstrate that hypoxic pre-conditioning decreases the activation of nuclear factor κB through the transcriptional induction of Disrupted in Schizophrenia-1.

  12. Nuclear Speckles

    PubMed Central

    Spector, David L.; Lamond, Angus I.

    2011-01-01

    Nuclear speckles, also known as interchromatin granule clusters, are nuclear domains enriched in pre-mRNA splicing factors, located in the interchromatin regions of the nucleoplasm of mammalian cells. When observed by immunofluorescence microscopy, they usually appear as 20–50 irregularly shaped structures that vary in size. Speckles are dynamic structures, and their constituents can exchange continuously with the nucleoplasm and other nuclear locations, including active transcription sites. Studies on the composition, structure, and dynamics of speckles have provided an important paradigm for understanding the functional organization of the nucleus and the dynamics of the gene expression machinery. PMID:20926517

  13. Importance of Hepatocyte Nuclear Factor 4α in Glycerol-induced Glucose-6-phosphatase Expression in Liver.

    PubMed

    Yoshida, Mitsuhiko; Lee, Eun Young; Kohno, Takashi; Tanaka, Tomoaki; Miyazaki, Masaru; Miki, Takashi

    2016-01-01

    Glucose-6-phosphatase (G6Pase) is a key regulator of gluconeogenesis. We previously found that administration of glycerol, a substrate for gluconeogenesis, transactivates G6Pase in the mouse liver. To clarify its cell-autonomous transcriptional activation in hepatocytes, we examined the mechanism of expression of the gene G6pc, which encodes G6Pase, in rat hepatoma cell line FAO cells. Endogenous G6pc expression in FAO cells was increased by glycerol administration as well as by the fatty acid oleate. Luciferase reporter assay revealed that the ~2.0 kb mouse G6pc promoter contains the element(s) responsible for glycerol-stimulated G6pc transactivation. Using several deletion- or chimeric-constructs of G6pc promoter, we found that the DNA response element for hepatocyte nuclear factor 4α (HNF4α) (-77/-65) in the G6pc promoter is essential for transactivation by glycerol. Similarly to glycerol, oleate also increased G6pc expression through its action on the HNF4α element (-77/-65). Furthermore, the reporter activities were higher in the cells co-treated with glycerol plus oleate than in those singly treated with glycerol or oleate. In addition, the temporal profiles of G6pc expression differed between glycerol and oleate administration. Our present results suggest that glycerol and oleate induce G6pc expression both via the HNF4αelement (-77/-65) and also through other regulatory mechanisms.

  14. Nuclear Science Division: 1993 Annual report

    SciTech Connect

    Myers, W.D.

    1994-06-01

    This report describes the activities of the Nuclear Science Division for the 1993 calendar year. This was another significant year in the history of the Division with many interesting and important accomplishments. Activities for the following programs are covered here: (1) nuclear structure and reactions program; (2) the Institute for Nuclear and Particle Astrophysics; (3) relativistic nuclear collisions program; (4) nuclear theory program; (5) nuclear data evaluation program, isotope project; and (6) 88-inch cyclotron operations.

  15. Heterodimeric interaction between retinoid X receptor alpha and orphan nuclear receptor OR1 reveals dimerization-induced activation as a novel mechanism of nuclear receptor activation.

    PubMed Central

    Wiebel, F F; Gustafsson, J A

    1997-01-01

    OR1 is a member of the steroid/thyroid hormone nuclear receptor superfamily which has been described to mediate transcriptional responses to retinoids and oxysterols. On a DR4 response element, an OR1 heterodimer with the nuclear receptor retinoid X receptor alpha (RXR alpha) has been described to convey transcriptional activation in both the absence and presence of the RXR ligand 9-cis retinoic acid, the mechanisms of which have remained unclear. Here, we dissect the effects of RXR alpha and OR1 ligand-binding domain interaction on transcriptional regulation and the role of the respective carboxy-terminal activation domains (AF-2s) in the absence and presence of the RXR ligand, employing chimeras of the nuclear receptors containing the heterologous GAL4 DNA-binding domain as well as natural receptors. The results show that the interaction of the RXR and OR1 ligand-binding domains unleashes a transcription activation potential that is mainly dependent on the AF-2 of OR1, indicating that interaction with RXR activates OR1. This defines dimerization-induced activation as a novel function of heterodimeric interaction and mechanism of receptor activation not previously described for nuclear receptors. Moreover, we present evidence that activation of OR1 occurs by a conformational change induced upon heterodimerization with RXR. PMID:9199332

  16. A Simple and Efficient Method to Detect Nuclear Factor Activation in Human Neutrophils by Flow Cytometry

    PubMed Central

    García-García, Erick; Uribe-Querol, Eileen; Rosales, Carlos

    2013-01-01

    Neutrophils are the most abundant leukocytes in peripheral blood. These cells are the first to appear at sites of inflammation and infection, thus becoming the first line of defense against invading microorganisms. Neutrophils possess important antimicrobial functions such as phagocytosis, release of lytic enzymes, and production of reactive oxygen species. In addition to these important defense functions, neutrophils perform other tasks in response to infection such as production of proinflammatory cytokines and inhibition of apoptosis. Cytokines recruit other leukocytes that help clear the infection, and inhibition of apoptosis allows the neutrophil to live longer at the site of infection. These functions are regulated at the level of transcription. However, because neutrophils are short-lived cells, the study of transcriptionally regulated responses in these cells cannot be performed with conventional reporter gene methods since there are no efficient techniques for neutrophil transfection. Here, we present a simple and efficient method that allows detection and quantification of nuclear factors in isolated and immunolabeled nuclei by flow cytometry. We describe techniques to isolate pure neutrophils from human peripheral blood, stimulate these cells with anti-receptor antibodies, isolate and immunolabel nuclei, and analyze nuclei by flow cytometry. The method has been successfully used to detect NF-κB and Elk-1 nuclear factors in nuclei from neutrophils and other cell types. Thus, this method represents an option for analyzing activation of transcription factors in isolated nuclei from a variety of cell types. PMID:23603868

  17. Nuclear activity and stellar population of a sample of interacting galaxies

    NASA Astrophysics Data System (ADS)

    Pastoriza, M. G.; Donzelli, C. J.; Bonatto, C.

    1999-07-01

    In this paper we investigate the nuclear activity and stellar population in a sample of 27 physical galaxy pairs. Equivalent widths of absorption features are used to characterise the nuclear stellar population according to templates: most galaxies of the sample have important flux contributions from stars younger than 10(8) years. According to classical diagnostic-diagrams the galaxies in our sample are either classified as H II regions or have emission line ratios near the transition zone between H II regions and LINERs. Based on the observed spectra, only 4 galaxies show LINER properties and 1 nucleus is a Seyfert 2. We found that the spectrum of a transition object (38% of the sample) can be described by a combination of an AGN with an H II region. As a result, 20 galaxies of the present sample may host a low-luminosity active nucleus. Based on observations made at CASLEO and CTIO. Complejo Astronómico El Leoncito (CASLEO) is operated under agreement between the Consejo Nacional de Investigaciones Cient\\'\\i ficas y Técnicas de la República Argentina and the National Universities of La Plata, Córdoba and San Juan.

  18. Nuclear Site Security in the Event of Terrorist Activity

    SciTech Connect

    Thomson, M.L.; Sims, J.

    2008-07-01

    This paper, presented as a poster, identifies why ballistic protection should now be considered at nuclear sites to counter terrorist threats. A proven and flexible form of multi purpose protection is described in detail with identification of trial results that show its suitability for this role. (authors)

  19. Phosphoinositide 3-Kinase Beta Protects Nuclear Envelope Integrity by Controlling RCC1 Localization and Ran Activity

    PubMed Central

    Redondo-Muñoz, Javier; Pérez-García, Vicente; Rodríguez, María J.; Valpuesta, José M.

    2014-01-01

    The nuclear envelope (NE) forms a barrier between the nucleus and the cytosol that preserves genomic integrity. The nuclear lamina and nuclear pore complexes (NPCs) are NE components that regulate nuclear events through interaction with other proteins and DNA. Defects in the nuclear lamina are associated with the development of laminopathies. As cells depleted of phosphoinositide 3-kinase beta (PI3Kβ) showed an aberrant nuclear morphology, we studied the contribution of PI3Kβ to maintenance of NE integrity. pik3cb depletion reduced the nuclear membrane tension, triggered formation of areas of lipid bilayer/lamina discontinuity, and impaired NPC assembly. We show that one mechanism for PI3Kβ regulation of NE/NPC integrity is its association with RCC1 (regulator of chromosome condensation 1), the activator of nuclear Ran GTPase. PI3Kβ controls RCC1 binding to chromatin and, in turn, Ran activation. These findings suggest that PI3Kβ regulates the nuclear envelope through upstream regulation of RCC1 and Ran. PMID:25348717

  20. [Retracted] Nuclear import of prototype foamy virus transactivator Bel1 is mediated by KPNA1, KPNA6 and KPNA7.

    PubMed

    Duan, Jihui; Tang, Zhiqin; Mu, Hong; Zhang, Guojun

    2017-03-01

    We would like to retract the article entitled "Nuclear import of prototype foamy virus transactivator Bel1 is mediated by KPNA1, KPNA6 and KPNA7" published in International Journal of Molecular Medicine 38: 339-406, 2016. The results presented in Fig. 6 have been demonstrated to be unreproducible, and we cannot provide an explanation for this. Furthermore, we have recently identified that the cell cultures used in our experiments were partly contaminated with Mycoplasma, which could have contributed to the irreproducibility of the results. In addition, we are currently in dispute with a colleague who has contributed towards this study, but does not wish to be included as a named author on the paper. We are therefore going to retract this article. All the authors unanimously agree to the retraction of this paper, and we deeply apologize to readers and editors for any inconvenience caused by this retraction. [the original article was published in the International Journal of Molecular Medicine 38: 399-406, 2016; DOI: 10.3892/ijmm.2016.2635].

  1. The methylation of the C-terminal region of hnRNPQ (NSAP1) is important for its nuclear localization

    SciTech Connect

    Passos, Dario O.; Quaresma, Alexandre J.C.; Kobarg, Joerg . E-mail: jkobarg@lnls.br

    2006-07-28

    Protein arginine methylation is an irreversible post-translational protein modification catalyzed by a family of at least nine different enzymes entitled PRMTs (protein arginine methyl transferases). Although PRMT1 is responsible for 85% of the protein methylation in human cells, its substrate spectrum has not yet been fully characterized nor are the functional consequences of methylation for the protein substrates well understood. Therefore, we set out to employ the yeast two-hybrid system in order to identify new substrate proteins for human PRMT1. We were able to identify nine different PRMT1 interacting proteins involved in different aspects of RNA metabolism, five of which had been previously described either as substrates for PRMT1 or as functionally associated with PRMT1. Among the four new identified possible protein substrates was hnRNPQ3 (NSAP1), a protein whose function has been implicated in diverse steps of mRNA maturation, including splicing, editing, and degradation. By in vitro methylation assays we were able to show that hnRNPQ3 is a substrate for PRMT1 and that its C-terminal RGG box domain is the sole target for methylation. By further studies with the inhibitor of methylation Adox we provide evidence that hnRNPQ1-3 are methylated in vivo. Finally, we demonstrate by immunofluorescence analysis of HeLa cells that the methylation of hnRNPQ is important for its nuclear localization, since Adox treatment causes its re-distribution from the nucleus to the cytoplasm.

  2. Molecular phylogeny of commercially important lobster species from Indian coast inferred from mitochondrial and nuclear DNA sequences.

    PubMed

    Jeena, N S; Gopalakrishnan, A; Radhakrishnan, E V; Kizhakudan, Joe K; Basheer, V S; Asokan, P K; Jena, J K

    2016-07-01

    Lobsters constitute low-volume high-value crustacean fishery resource along Indian coast. For the conservation and management of this declining resource, accurate identification of species and larvae is essential. The objectives of this work were to generate species-specific molecular signatures of 11 commercially important species of lobsters of families Palinuridae and Scyllaridae and to reconstruct a phylogeny to clarify the evolutionary relationships among genera and species included in this study. Partial sequences were generated for all the candidate species from sampling sites along the Indian coast using markers like Cytochrome oxidase I (COI), 16SrRNA, 12SrRNA, and 18SrRNA genes, and analyzed. The genetic identities of widely distributed Thenus species along the Indian coast to be Thenus unimaculatus and the sub-species of Panulirus homarus to be P. homarus homarus were confirmed. Phylogeny reconstruction using the individual gene and concatenated mtDNA data set were carried out. The overall results suggested independent monophyly of Scyllaridae and Stridentes of Palinuridae. The interspecific divergence was found to be highest for the 12SrRNA compared with other genes. Significant incongruence between mtDNA and nuclear 18SrRNA gene tree topologies was observed. The results hinted an earlier origin for Palinuridae compared with Scyllaridae. The DNA sequence data generated from this study will aid in the correct identification of lobster larvae and will find application in research related to larval transport and distribution.

  3. Nuclear interactor of ARF and Mdm2 regulates multiple pathways to activate p53

    PubMed Central

    Reed, Sara M; Hagen, Jussara; Tompkins, Van S; Thies, Katie; Quelle, Frederick W; Quelle, Dawn E

    2014-01-01

    The p53 tumor suppressor is controlled by an interactive network of factors that stimulate or inhibit its transcriptional activity. Within that network, Mdm2 functions as the major antagonist of p53 by promoting its ubiquitylation and degradation. Conversely, Tip60 activates p53 through direct association on target promoters as well as acetylation of p53 at lysine 120 (K120). This study examines the functional relationship between Mdm2 and Tip60 with a novel p53 regulator, NIAM (nuclear interactor of ARF and Mdm2). Previous work showed NIAM can suppress proliferation and activate p53 independently of ARF, indicating that other factors mediate those activities. Here, we demonstrate that NIAM is a chromatin-associated protein that binds Tip60. NIAM can promote p53 K120 acetylation, although that modification is not required for NIAM to inhibit proliferation or induce p53 transactivation of the p21 promoter. Notably, Tip60 silencing showed it contributes to but is not sufficient for NIAM-mediated p53 activation, suggesting other mechanisms are involved. Indeed, growth-inhibitory forms of NIAM also bind to Mdm2, and increased NIAM expression levels disrupt p53–Mdm2 association, inhibit p53 polyubiquitylation, and prevent Mdm2-mediated inhibition of p53 transcriptional activity. Importantly, loss of NIAM significantly impairs p53 activation. Together, these results show that NIAM activates p53 through multiple mechanisms involving Tip60 association and Mdm2 inhibition. Thus, NIAM regulates 2 critical pathways that control p53 function and are altered in human cancers, implying an important role for NIAM in tumorigenesis. PMID:24621507

  4. Strontium-90 activity concentration in soil samples from the exclusion zone of the Fukushima daiichi nuclear power plant

    PubMed Central

    Sahoo, Sarata Kumar; Kavasi, Norbert; Sorimachi, Atsuyuki; Arae, Hideki; Tokonami, Shinji; Mietelski, Jerzy Wojciech; Łokas, Edyta; Yoshida, Satoshi

    2016-01-01

    The radioactive fission product 90Sr has a long biological half-life (˜18 y) in the human body. Due to its chemical similarity to calcium it accumulates in bones and irradiates the bone marrow, causing its high radio-toxicity. Assessing 90Sr is therefore extremely important in case of a nuclear disaster. In this work 16 soil samples were collected from the exclusion zone (<30 km) of the earthquake-damaged Fukushima Daiichi nuclear power plant, to measure 90Sr activity concentration using liquid scintillation counting. 137Cs activity concentration was also measured with gamma-spectroscopy in order to investigate correlation with 90Sr. The 90Sr activity concentrations ranged from 3.0 ± 0.3 to 23.3 ± 1.5 Bq kg−1 while the 137Cs from 0.7 ± 0.1 to 110.8 ± 0.3 kBq kg−1. The fact that radioactive contamination originated from the Fukushima nuclear accident was obvious due to the presence of 134Cs. However, 90Sr contamination was not confirmed in all samples although detectable amounts of 90Sr can be expected in Japanese soils, as a background, stemming from global fallout due to the atmospheric nuclear weapon tests. Correlation analysis between 90Sr and 137Cs activity concentrations provides a potentially powerful tool to discriminate background 90Sr level from its Fukushima contribution. PMID:27048779

  5. Strontium-90 activity concentration in soil samples from the exclusion zone of the Fukushima daiichi nuclear power plant.

    PubMed

    Sahoo, Sarata Kumar; Kavasi, Norbert; Sorimachi, Atsuyuki; Arae, Hideki; Tokonami, Shinji; Mietelski, Jerzy Wojciech; Łokas, Edyta; Yoshida, Satoshi

    2016-04-06

    The radioactive fission product (90)Sr has a long biological half-life (˜18 y) in the human body. Due to its chemical similarity to calcium it accumulates in bones and irradiates the bone marrow, causing its high radio-toxicity. Assessing (90)Sr is therefore extremely important in case of a nuclear disaster. In this work 16 soil samples were collected from the exclusion zone (<30 km) of the earthquake-damaged Fukushima Daiichi nuclear power plant, to measure (90)Sr activity concentration using liquid scintillation counting. (137)Cs activity concentration was also measured with gamma-spectroscopy in order to investigate correlation with (90)Sr. The (90)Sr activity concentrations ranged from 3.0 ± 0.3 to 23.3 ± 1.5 Bq kg(-1) while the (137)Cs from 0.7 ± 0.1 to 110.8 ± 0.3 kBq kg(-1). The fact that radioactive contamination originated from the Fukushima nuclear accident was obvious due to the presence of (134)Cs. However, (90)Sr contamination was not confirmed in all samples although detectable amounts of (90)Sr can be expected in Japanese soils, as a background, stemming from global fallout due to the atmospheric nuclear weapon tests. Correlation analysis between (90)Sr and (137)Cs activity concentrations provides a potentially powerful tool to discriminate background (90)Sr level from its Fukushima contribution.

  6. Strontium-90 activity concentration in soil samples from the exclusion zone of the Fukushima daiichi nuclear power plant

    NASA Astrophysics Data System (ADS)

    Sahoo, Sarata Kumar; Kavasi, Norbert; Sorimachi, Atsuyuki; Arae, Hideki; Tokonami, Shinji; Mietelski, Jerzy Wojciech; Łokas, Edyta; Yoshida, Satoshi

    2016-04-01

    The radioactive fission product 90Sr has a long biological half-life (˜18 y) in the human body. Due to its chemical similarity to calcium it accumulates in bones and irradiates the bone marrow, causing its high radio-toxicity. Assessing 90Sr is therefore extremely important in case of a nuclear disaster. In this work 16 soil samples were collected from the exclusion zone (<30 km) of the earthquake-damaged Fukushima Daiichi nuclear power plant, to measure 90Sr activity concentration using liquid scintillation counting. 137Cs activity concentration was also measured with gamma-spectroscopy in order to investigate correlation with 90Sr. The 90Sr activity concentrations ranged from 3.0 ± 0.3 to 23.3 ± 1.5 Bq kg‑1 while the 137Cs from 0.7 ± 0.1 to 110.8 ± 0.3 kBq kg‑1. The fact that radioactive contamination originated from the Fukushima nuclear accident was obvious due to the presence of 134Cs. However, 90Sr contamination was not confirmed in all samples although detectable amounts of 90Sr can be expected in Japanese soils, as a background, stemming from global fallout due to the atmospheric nuclear weapon tests. Correlation analysis between 90Sr and 137Cs activity concentrations provides a potentially powerful tool to discriminate background 90Sr level from its Fukushima contribution.

  7. 10 CFR 73.55 - Requirements for physical protection of licensed activities in nuclear power reactors against...

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... activities in nuclear power reactors against radiological sabotage. 73.55 Section 73.55 Energy NUCLEAR... power reactors against radiological sabotage. (a) Introduction. (1) By March 31, 2010, each nuclear... this section as applicable to operating nuclear power reactor facilities. (6) Applicants for...

  8. 10 CFR 73.55 - Requirements for physical protection of licensed activities in nuclear power reactors against...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... activities in nuclear power reactors against radiological sabotage. 73.55 Section 73.55 Energy NUCLEAR... power reactors against radiological sabotage. (a) Introduction. (1) By March 31, 2010, each nuclear... this section as applicable to operating nuclear power reactor facilities. (6) Applicants for...

  9. Phosphorylated nuclear receptor CAR forms a homodimer to repress its constitutive activity for ligand activation.

    PubMed

    Shizu, Ryota; Osabe, Makoto; Perera, Lalith; Moore, Rick; Sueyoshi, Tatsuya; Negishi, Masahiko

    2017-03-06

    Nuclear receptor CAR (NR1I3) regulates hepatic drug and energy metabolism as well as cell fate. Its activation can be a critical factor in drug-induced toxicity and disease development such as diabetes and tumors. CAR inactivates its constitutive activity by phosphorylation at threonine 38. Utilizing receptor for protein kinase 1 (RACK1) as the regulatory subunit, protein phosphatase PP2A dephosphorylates threonine 38 to activate CAR. Here we have demonstrated that CAR undergoes its homodimer-monomer conversion to regulate this dephosphorylation. By co-expressing two differently-tagged CAR proteins in Huh-7 cells, mouse primary hepatocytes and mouse livers, co-immunoprecipitation and two-dimensional gel electrophoresis revealed that CAR can form a homodimer in a configuration in which the PP2A/RACK1 binding site is buried within its dimer interface. Epidermal growth factor (EGF) was found to stimulate CAR homo-dimerization, thus constraining CAR in its inactive form. The agonistic ligand CITCO binds directly to the CAR homodimer and dissociates phosphorylated CAR into its monomer, exposing the PP2A/RACK1 binding site for dephosphorylation. Phenobarbital, which is not a CAR ligand, binds the EGF receptor, reversing the EGF signal to monomerize CAR for its indirect activation. Thus, the homodimer-monomer conversion is the underlying molecular mechanism that regulates CAR activation, by placing phosphorylated threonine 38 as the common target for both direct and in direct activation of CAR.

  10. Phosphatidylinositol 3-kinase/Akt signaling enhances nuclear localization and transcriptional activity of BRCA1

    SciTech Connect

    Hinton, Cimona V.; Fitzgerald, Latricia D.; Thompson, Marilyn E. . E-mail: methompson@mmc.edu

    2007-05-15

    Signaling pathways involved in regulating nuclear-cytoplasmic distribution of BRCA1 have not been previously reported. Here, we provide evidence that heregulin {beta}1-induced activation of the Akt pathway increases the nuclear content of BRCA1. First, treatment of T47D breast cancer cells with heregulin {beta}1 results in a two-fold increase in nuclear BRCA1 as assessed by FACS analysis, immunoblotting and immunofluorescence. This heregulin-induced increase in nuclear BRCA1 is blocked by siRNA-mediated down-regulation of Akt. Second, mutation of threonine 509 in BRCA1, the site of Akt phosphorylation, to an alanine, attenuates the ability of heregulin to induce BRCA1 nuclear accumulation. These data suggest that Akt-catalyzed phosphorylation of BRCA1 is required for the heregulin-regulated nuclear concentration of BRCA1. Because most functions ascribed to BRCA1 occur within the nucleus, we postulated that phosphorylation-dependent nuclear accumulation of BRCA1 would result in enhanced nuclear activity, specifically transcriptional activity, of BRCA1. This postulate is affirmed by our observation that the ability of BRCA1 to transactivate GADD45 promoter constructs was enhanced in T47D cells treated with heregulin {beta}1. Furthermore, the heterologous expression of BRCA1 in HCC1937 human breast cancer cells, which have constitutively active Akt, also induces GADD45 promoter activity, whereas the expression of BRCA1 in which threonine 509 has been mutated to an alanine is able to only minimally induce promoter activity. These findings implicate Akt in upstream events leading to BRCA1 nuclear localization and function.

  11. 76 FR 40749 - Agency Information Collection Activities: Records and Supporting Data: Importation, Receipt...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-11

    ... of Alcohol, Tobacco, Firearms and Explosives Agency Information Collection Activities: Records and Supporting Data: Importation, Receipt, Storage, and Disposition by Explosives Importers, Manufacturers..., Firearms and Explosives (ATF), will be submitting the following information collection request to...

  12. Nuclear Thermal Rocket Element Environmental Simulator (NTREES) Upgrade Activities

    NASA Technical Reports Server (NTRS)

    Emrich, William

    2013-01-01

    A key technology element in Nuclear Thermal Propulsion is the development of fuel materials and components which can withstand extremely high temperatures while being exposed to flowing hydrogen. NTREES provides a cost effective method for rapidly screening of candidate fuel components with regard to their viability for use in NTR systems. The NTREES is designed to mimic the conditions (minus the radiation) to which nuclear rocket fuel elements and other components would be subjected to during reactor operation. The NTREES consists of a water cooled ASME code stamped pressure vessel and its associated control hardware and instrumentation coupled with inductive heaters to simulate the heat provided by the fission process. The NTREES has been designed to safely allow hydrogen gas to be injected into internal flow passages of an inductively heated test article mounted in the chamber.

  13. Robustness of Nuclear Core Activity Reconstruction by Data Assimilation

    NASA Astrophysics Data System (ADS)

    Bouriquet, Bertrand; Argaud, Jean-Philippe; Erhard, Patrick; Massart, Sébastien; Ponçot, Angélique; Ricci, Sophie; Thual, Olivier

    Inspired from meteorological applications, data assimilation techniques can be used to perform an optimal reconstruction of the neutronic field in a nuclear reactor core. Measurements and simulation information, coming from a numerical model, are merged together to build a better estimation of the whole field. In this paper, we first study the robustness of the method when the amount of measured information varies. We then study the influence of the nature of the instruments and their spatial repartition on the efficiency of the field reconstruction. This study also highlights the instruments providing most information within a data assimilation procedure. The study of various network configurations of instruments in the nuclear core establishes that the influence of each instrument depends both on the individual instrumentation location as well as on the chosen network.

  14. Nuclear factor of activated T cells (NFAT) signaling regulates PTEN expression and intestinal cell differentiation

    PubMed Central

    Wang, Qingding; Zhou, Yuning; Jackson, Lindsey N.; Johnson, Sara M.; Chow, Chi-Wing; Evers, B. Mark

    2011-01-01

    The nuclear factor of activated T cell (NFAT) proteins are a family of transcription factors (NFATc1–c4) involved in the regulation of cell differentiation and adaptation. Previously we demonstrated that inhibition of phosphatidylinositol 3-kinase or overexpression of PTEN enhanced intestinal cell differentiation. Here we show that treatment of intestinal-derived cells with the differentiating agent sodium butyrate (NaBT) increased PTEN expression, NFAT binding activity, and NFAT mRNA expression, whereas pretreatment with the NFAT signaling inhibitor cyclosporine A (CsA) blocked NaBT-mediated PTEN induction. Moreover, knockdown of NFATc1 or NFATc4, but not NFATc2 or NFATc3, attenuated NaBT-induced PTEN expression. Knockdown of NFATc1 decreased PTEN expression and increased the phosphorylation levels of Akt and downstream targets Foxo1 and GSK-3α/β. Furthermore, overexpression of NFATc1 or the NFATc4 active mutant increased PTEN and p27kip1 expression and decreased Akt phosphorylation. In addition, pretreatment with CsA blocked NaBT-mediated induction of intestinal alkaline phosphatase (IAP) activity and villin and p27kip1 expression; knockdown of either NFATc1 or NFATc4 attenuated NaBT-induced IAP activity. We provide evidence showing that NFATc1 and NFATc4 are regulators of PTEN expression. Importantly, our results suggest that NFATc1 and NFATc4 regulation of intestinal cell differentiation may be through PTEN regulation. PMID:21148296

  15. Comparison of the activity measurements in nuclear medicine services in the Brazilian northeast region.

    PubMed

    de Farias Fragoso, Maria da Conceição; de Albuquerque, Antônio Morais; de Oliveira, Mércia L; de Lima, Fabiana Farias; Barreto, Flávio Chiappetta Paes; de Andrade Lima, Ricardo

    2013-12-01

    The Northeastern Regional Centre for Nuclear Sciences (CRCN-NE), National Nuclear Energy Commission, has organized for the first time in nuclear medicine services (NMSs) in the Brazilian northeast region a comparison of activity measurements for (99m)Tc, (131)I, (67)Ga, (201)Tl and (57)Co. This tool is widely utilized to evaluate not only the accuracy of radionuclide calibrators, but also the competence of NMSs to measure the activity of the radiopharmaceuticals and the performance of the personnel involved in these measurements. The comparison results showed that 90% of the results received from participants are within the ±10% limit established by the Brazilian Norm.

  16. Nuclear import of high risk HPV16 E7 oncoprotein is mediated by its zinc-binding domain via hydrophobic interactions with Nup62

    PubMed Central

    Eberhard, Jeremy; Onder, Zeynep; Moroianu, Junona

    2013-01-01

    We previously discovered that nuclear import of high risk HPV16 E7 is mediated by a cNLS located within the zinc-binding domain via a pathway that is independent of karyopherins/importins (Angeline et al., 2003; Knapp et al., 2009). In this study we continued our characterization of the cNLS and nuclear import pathway of HPV16 E7. We find that an intact zinc-binding domain is essential for the cNLS function in mediating nuclear import of HPV16 E7. Mutagenesis of cysteine residues to alanine in each of the two CysXXCys motifs involved in zinc-binding changes the nuclear localization of the EGFP-16E7 and 2xEGFP-16E7 mutants. We further discover that a patch of hydrophobic residues, 65LRLCV69, within the zinc-binding domain of HPV16 E7 mediates its nuclear import via hydrophobic interactions with the FG domain of the central channel nucleoporin Nup62. PMID:24074597

  17. Nuclear Import of the Thyroid Hormone Receptor α1 is Mediated by Importin 7, Importin β1, and Adaptor Importin α1

    PubMed Central

    Roggero, Vincent R.; Zhang, Jibo; Parente, Laura E.; Doshi, Yazdi; Dziedzic, Rose C.; McGregor, Emma L.; Varjabedian, Arev D.; Schad, Sara E.; Bondzi, Cornelius; Allison, Lizabeth A.

    2015-01-01

    The thyroid hormone receptor α1 (TRα1) is a nuclear receptor for thyroid hormone that shuttles rapidly between the nucleus and cytoplasm. Our prior studies showed that nuclear import of TRα1 is directed by two nuclear localization signals, one in the N-terminal A/B domain and the other in the hinge domain. Here, we showed using in vitro nuclear import assays that TRα1 nuclear localization is temperature and energy-dependent and can be reconstituted by the addition of cytosol. In HeLa cells expressing green fluorescent protein (GFP)-tagged TRα1, knockdown of importin 7, importin β1 and importin α1 by RNA interference, or treatment with an importin β1-specific inhibitor, significantly reduced nuclear localization of TRα1, while knockdown of other importins had no effect. Coimmunoprecipitation assays confirmed that TRα1 interacts with importin 7, as well as importin β1 and the adapter importin α1, suggesting that TRα1 trafficking into the nucleus is mediated by two distinct pathways. PMID:26525414

  18. Human Immunodeficiency Virus Type 1 (HIV-1) Induces the Cytoplasmic Retention of Heterogeneous Nuclear Ribonucleoprotein A1 by Disrupting Nuclear Import

    PubMed Central

    Monette, Anne; Ajamian, Lara; López-Lastra, Marcelo; Mouland, Andrew J.

    2009-01-01

    Human immunodeficiency virus type 1 (HIV-1) co-opts host proteins and cellular machineries to its advantage at every step of the replication cycle. Here we show that HIV-1 enhances heterogeneous nuclear ribonucleoprotein (hnRNP) A1 expression and promotes the relocalization of hnRNP A1 to the cytoplasm. The latter was dependent on the nuclear export of the unspliced viral genomic RNA (vRNA) and to alterations in the abundance and localization of the FG-repeat nuclear pore glycoprotein p62. hnRNP A1 and vRNA remain colocalized in the cytoplasm supporting a post-nuclear function during the late stages of HIV-1 replication. Consistently, we show that hnRNP A1 acts as an internal ribosomal entry site trans-acting factor up-regulating internal ribosome entry site-mediated translation initiation of the HIV-1 vRNA. The up-regulation and cytoplasmic retention of hnRNP A1 by HIV-1 would ensure abundant expression of viral structural proteins in cells infected with HIV-1. PMID:19737937

  19. Mitogen-Activated Protein Kinase Activity Is Not Essential for the First Step of Nuclear Reprogramming in Bovine Somatic Cell Nuclear Transfer.

    PubMed

    Tani, Tetsuya; Kato, Yoko

    2017-03-07

    For reprogramming a somatic nucleus during mammalian cloning, metaphase of the second meiotic division (MII) oocytes has been widely used as recipient cytoplasm. High activity of maturation-promoting factor (MPF) and mitogen-activated protein kinase (MAPK) is believed to accelerate the remodeling and/or reprogramming of a somatic nucleus introduced into the ooplasm by somatic cell nuclear transfer. We demonstrated previously that the first step in nuclear reprogramming is not directly regulated by MPF and MAPK because activated oocytes in which MPF activity is diminished and MAPK activity is maintained can develop to the blastocyst stage after receiving an M phase somatic nucleus in bovine cloning. In this study, our aim was to test whether MAPK activity is necessary for the first step in nuclear reprogramming and/or chromatin remodeling (phosphorylation of histone H3 at Ser3, trimethylation of histone H3 at Lys 9, and acetylation of histone H3 at Lys14) in bovine somatic cloning. We found that it was not necessary, and neither was MPF activity.

  20. 10 CFR 73.74 - Requirement for advance notice and protection of import shipments of nuclear material from...

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Nuclear Material (i.e., not listed in appendix F of this part) shall: (1) Notify in writing the Director... significance, or irradiated reactor fuel shall assure that during transport outside the United States...

  1. 10 CFR 73.74 - Requirement for advance notice and protection of import shipments of nuclear material from...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Nuclear Material (i.e., not listed in appendix F of this part) shall: (1) Notify in writing the Director... significance, or irradiated reactor fuel shall assure that during transport outside the United States...

  2. Novel interaction between nuclear co-activator CBP and the CDK5 activator binding protein - C53.

    PubMed

    Yin, Xiaolong; Warner, Dennis R; Roberts, Emily A; Pisano, M Michele; Greene, Robert M

    2005-08-01

    cAMP response element-binding protein (CREB)-binding protein (CBP) is a multifunctional transcriptional co-activator that plays important roles in cell proliferation and differentiation. CBP is expressed in murine embryonic orofacial tissue and is developmentally regulated. To identify nuclear factors associating with CBP in developing orofacial tissue, a yeast two-hybrid screen of a cDNA library derived from embryonic orofacial tissue from gestational days 11-13 mouse embryos was conducted. The carboxy terminal region of CBP (including the C/H3 region) was utilized as a bait. C53, a 57 kDa protein known to bind to the p25 activator of cyclin-dependent kinase 5, was identified as a novel binding partner of CBP. The association of C53 with CBP was confirmed in vitro by glutathione S-transferase pull-down assays, and in vivo by co-immunoprecipitation. Reporter assays demonstrated that C53 had little effect on CBP mediated transcriptional activation. These results identify C53 as a novel binding partner for CBP. Recent research on presenilin-loss induced neurodegeneration demonstrated decreased expression of CBP and increased levels of the Cdk5 activator p25, both C53 binding proteins, suggesting that C53 might play a role in regulating neuronal proliferation, migration and/or differentiation in embryonic development.

  3. Nuclear Import and Dimerization of Tomato ASR1, a Water Stress-Inducible Protein Exclusive to Plants

    PubMed Central

    Ricardi, Martiniano M.; Guaimas, Francisco F.; González, Rodrigo M.; Burrieza, Hernán P.; López-Fernández, María P.; Estévez, José M.; Iusem, Norberto D.

    2012-01-01

    The ASR (for ABA/water stress/ripening) protein family, first described in tomato as nuclear and involved in adaptation to dry climates, is widespread in the plant kingdom, including crops of high agronomic relevance. We show both nuclear and cytosolic localization for ASR1 (the most studied member of the family) in histological plant samples by immunodetection, typically found in small proteins readily diffusing through nuclear pores. Indeed, a nuclear localization was expected based on sorting prediction software, which also highlight a monopartite nuclear localization signal (NLS) in the primary sequence. However, here we prove that such an “NLS” of ASR1 from tomato is dispensable and non-functional, being the transport of the protein to the nucleus due to simple diffusion across nuclear pores. We attribute such a targeting deficiency to the misplacing in that cryptic NLS of two conserved contiguous lysine residues. Based on previous in vitro experiments regarding quaternary structure, we also carried out live cell imaging assays through confocal microscopy to explore dimer formation in planta. We found homodimers in both the cytosol and the nucleus and demonstrated that assembly of both subunits together can occur in the cytosol, giving rise to translocation of preformed dimers. The presence of dimers was further corroborated by means of in vivo crosslinking of nuclei followed by SDS-PAGE. PMID:22899993

  4. Nuclear import and dimerization of tomato ASR1, a water stress-inducible protein exclusive to plants.

    PubMed

    Ricardi, Martiniano M; Guaimas, Francisco F; González, Rodrigo M; Burrieza, Hernán P; López-Fernández, María P; Jares-Erijman, Elizabeth A; Estévez, José M; Iusem, Norberto D

    2012-01-01

    The ASR (for ABA/water stress/ripening) protein family, first described in tomato as nuclear and involved in adaptation to dry climates, is widespread in the plant kingdom, including crops of high agronomic relevance. We show both nuclear and cytosolic localization for ASR1 (the most studied member of the family) in histological plant samples by immunodetection, typically found in small proteins readily diffusing through nuclear pores. Indeed, a nuclear localization was expected based on sorting prediction software, which also highlight a monopartite nuclear localization signal (NLS) in the primary sequence. However, here we prove that such an "NLS" of ASR1 from tomato is dispensable and non-functional, being the transport of the protein to the nucleus due to simple diffusion across nuclear pores. We attribute such a targeting deficiency to the misplacing in that cryptic NLS of two conserved contiguous lysine residues. Based on previous in vitro experiments regarding quaternary structure, we also carried out live cell imaging assays through confocal microscopy to explore dimer formation in planta. We found homodimers in both the cytosol and the nucleus and demonstrated that assembly of both subunits together can occur in the cytosol, giving rise to translocation of preformed dimers. The presence of dimers was further corroborated by means of in vivo crosslinking of nuclei followed by SDS-PAGE.

  5. (129)I record of nuclear activities in marine sediment core from Jiaozhou Bay in China.

    PubMed

    Fan, Yukun; Hou, Xiaolin; Zhou, Weijian; Liu, Guangshan

    2016-04-01

    Iodine-129 has been used as a powerful tool for environmental tracing of human nuclear activities. In this work, a sediment core collected from Jiaozhou Bay, the east coast of China, in 2002 was analyzed for (129)I to investigate the influence of human nuclear activities in this region. Significantly enhanced (129)I level was observed in upper 70 cm of the sediment core, with peak values in the layer corresponding to 1957, 1964, 1974, 1986, and after 1990. The sources of (129)I and corresponding transport processes in this region are discussed, including nuclear weapons testing at the Pacific Proving Grounds, global fallout from a large numbers of nuclear weapon tests in 1963, the climax of Chinese nuclear weapons testing in the early 1970s, the Chernobyl accident in 1986, and long-distance dispersion of European reprocessing derived (129)I. The very well (129)I records of different human nuclear activities in the sediment core illustrate the potential application of (129)I in constraining ages and sedimentation rates of the recent sediment. The releases of (129)I from the European nuclear fuel reprocessing plants at La Hague (France) and Sellafield (UK) were found to dominate the inventory of (129)I in the Chinese sediments after 1990, not only the directly atmospheric releases of these reprocessing plants, but also re-emission of marine discharged (129)I of these reprocessing plants in the highly contaminated European seas.

  6. Technosocial Modeling for Determining the Status and Nature of a State’s Nuclear Activities

    SciTech Connect

    Gastelum, Zoe N.; Harvey, Julia B.

    2009-09-25

    The International Atomic Energy Agency State Evaluation Process: The Role of Information Analysis in Reaching Safeguards Conclusions (Mathews et al. 2008), several examples of nonproliferation models using analytical software were developed that may assist the IAEA with collecting, visualizing, analyzing, and reporting information in support of the State Evaluation Process. This paper focuses on one of the examples a set of models developed in the Proactive Scenario Production, Evidence Collection, and Testing (ProSPECT) software that evaluates the status and nature of a state’s nuclear activities. The models use three distinct subject areas to perform this assessment: the presence of nuclear activities, the consistency of those nuclear activities with national nuclear energy goals, and the geopolitical context in which those nuclear activities are taking place. As a proof-of-concept for the models, a crude case study was performed. The study, which attempted to evaluate the nuclear activities taking place in Syria prior to September 2007, yielded illustrative, yet inconclusive, results. Due to the inconclusive nature of the case study results, changes that may improve the model’s efficiency and accuracy are proposed.

  7. Activities in support of continuing the service of nuclear power plant concrete structures

    SciTech Connect

    Naus, Dan J

    2012-01-01

    In general, nuclear power plant concrete structure s performance has been very good; however, aging of concrete structures occurs with the passage of time that can potentially result in degradation if is effects are not controlled. Safety-related nuclear power plant concrete structures are described. In-service inspection and testing requirements in the U.S. are summarized. The interaction of the license renewal process and concrete structures is noted. A summary of operating experience related to aging of nuclear power plant concrete structures is provided. Several candidate areas are identified where additional research would be beneficial for aging management of nuclear power plant concrete structures. Finally, an update on recent activities at Oak Ridge National Laboratory related to aging management of nuclear power plant concrete structures is provided.

  8. Using Importance-Performance Analysis to Guide Instructional Design of Experiential Learning Activities

    ERIC Educational Resources Information Center

    Anderson, Sheri; Hsu, Yu-Chang; Kinney, Judy

    2016-01-01

    Designing experiential learning activities requires an instructor to think about what they want the students to learn. Using importance-performance analysis can assist with the instructional design of the activities. This exploratory study used importance-performance analysis in an online introduction to criminology course. There is limited…

  9. Dietary regulation of adiponectin by direct and indirect lipid activators of nuclear hormone receptors.

    PubMed

    Rühl, R; Landrier, J F

    2016-01-01

    Adiponectin is an adipokine mainly secreted by adipocytes that presents antidiabetic, anti-inflammatory, and antiatherogenic functions. Therefore, modulation of adiponectin expression represents a promising target for prevention or treatment of several diseases including insulin resistance and type II diabetes. Pharmacological agents such as the nuclear hormone receptor synthetic agonists like peroxisome proliferator activated receptor γ agonists are of particular interest in therapeutic strategies due to their ability to increase the plasma adiponectin concentration. Nutritional approaches are also of particular interest, especially in primary prevention, since some active compounds of our diet (notably vitamins, carotenoids, or other essential nutrients) are direct or indirect lipid-activators of nuclear hormone receptors and are modifiers of adiponectin expression and secretion. The aim of the present review is to summarize current knowledge about the nutritional regulation of adiponectin by derivatives of active compounds naturally present in the diet acting as indirect or direct activators of nuclear hormone receptors.

  10. 77 FR 70846 - Regulatory Guide 1.182, “Assessing and Managing Risk Before Maintenance Activities at Nuclear...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-27

    ... Chief, Regulatory Guide Development Branch, Division of Engineering, Office of Nuclear Regulatory... From the Federal Register Online via the Government Publishing Office NUCLEAR REGULATORY COMMISSION Regulatory Guide 1.182, ``Assessing and Managing Risk Before Maintenance Activities at...

  11. THE AIMS AND ACTIVITIES OF THE INTERNATIONAL NETWORK OF NUCLEAR STRUCTURE AND DECAY DATA EVALUATORS.

    SciTech Connect

    NICHOLS,A.L.; TULI, J.K.

    2007-04-22

    International Network of Nuclear Structure and Decay Data (NSDD) Evaluators consists of a number of evaluation groups and data service centers in several countries that appreciate the merits of working together to maintain and ensure the quality and comprehensive content of the ENSDF database (Evaluated Nuclear Structure Data File). Biennial meetings of the network are held under the auspices of the International Atomic Energy Agency (IAEA) to assign evaluation responsibilities, monitor progress, discuss improvements and emerging difficulties, and agree on actions to be undertaken by individual members. The evaluated data and bibliographic details are made available to users via various media, such as the journals ''Nuclear Physics A'' and ''Nuclear Data Sheets'', the World Wide Web, on CD-ROM, wall charts of the nuclides and ''Nuclear Wallet Cards''. While the ENSDF master database is maintained by the US National Nuclear Data Center at the Brookhaven National Laboratory, these data are also available from other nuclear data centers including the IAEA Nuclear Data Section. The Abdus Salam International Centre for Theoretical Physics (ICTP), Trieste, Italy, in cooperation with the IAEA, organizes workshops on NSDD at regular intervals. The primary aims of these particular workshops are to provide hands-on training in the data evaluation processes, and to encourage new evaluators to participate in NSDD activities. The technical contents of these NSDD workshops are described, along with the rationale for the inclusion of various topics.

  12. Nuclear cereblon modulates transcriptional activity of Ikaros and regulates its downstream target, enkephalin, in human neuroblastoma cells.

    PubMed

    Wada, Takeyoshi; Asahi, Toru; Sawamura, Naoya

    2016-08-26

    The gene coding cereblon (CRBN) was originally identified in genetic linkage analysis of mild autosomal recessive nonsyndromic intellectual disability. CRBN has broad localization in both the cytoplasm and nucleus. However, the significance of nuclear CRBN remains unknown. In the present study, we aimed to elucidate the role of CRBN in the nucleus. First, we generated a series of CRBN deletion mutants and determined the regions responsible for the nuclear localization. Only CRBN protein lacking the N-terminal region was localized outside of the nucleus, suggesting that the N-terminal region is important for its nuclear localization. CRBN was also identified as a thalidomide-binding protein and component of the cullin-4-containing E3 ubiquitin ligase complex. Thalidomide has been reported to be involved in the regulation of the transcription factor Ikaros by CRBN-mediated degradation. To investigate the nuclear functions of CRBN, we performed co-immunoprecipitation experiments and evaluated the binding of CRBN to Ikaros. As a result, we found that CRBN was associated with Ikaros protein, and the N-terminal region of CRBN was required for Ikaros binding. In luciferase reporter gene experiments, CRBN modulated transcriptional activity of Ikaros. Furthermore, we found that CRBN modulated Ikaros-mediated transcriptional repression of the proenkephalin gene by binding to its promoter region. These results suggest that CRBN binds to Ikaros via its N-terminal region and regulates transcriptional activities of Ikaros and its downstream target, enkephalin.

  13. Nuclear disarmament verification

    SciTech Connect

    DeVolpi, A.

    1993-12-31

    Arms control treaties, unilateral actions, and cooperative activities -- reflecting the defusing of East-West tensions -- are causing nuclear weapons to be disarmed and dismantled worldwide. In order to provide for future reductions and to build confidence in the permanency of this disarmament, verification procedures and technologies would play an important role. This paper outlines arms-control objectives, treaty organization, and actions that could be undertaken. For the purposes of this Workshop on Verification, nuclear disarmament has been divided into five topical subareas: Converting nuclear-weapons production complexes, Eliminating and monitoring nuclear-weapons delivery systems, Disabling and destroying nuclear warheads, Demilitarizing or non-military utilization of special nuclear materials, and Inhibiting nuclear arms in non-nuclear-weapons states. This paper concludes with an overview of potential methods for verification.

  14. Nuclear envelope lamin-A couples actin dynamics with immunological synapse architecture and T cell activation.

    PubMed

    González-Granado, José M; Silvestre-Roig, Carlos; Rocha-Perugini, Vera; Trigueros-Motos, Laia; Cibrián, Danay; Morlino, Giulia; Blanco-Berrocal, Marta; Osorio, Fernando G; Freije, José M P; López-Otín, Carlos; Sánchez-Madrid, Francisco; Andrés, Vicente

    2014-04-22

    In many cell types, nuclear A-type lamins regulate multiple cellular functions, including higher-order genome organization, DNA replication and repair, gene transcription, and signal transduction; however, their role in specialized immune cells remains largely unexplored. We showed that the abundance of A-type lamins was almost negligible in resting naïve T lymphocytes, but was increased upon activation of the T cell receptor (TCR). The increase in lamin-A was an early event that accelerated formation of the immunological synapse between T cells and antigen-presenting cells. Polymerization of F-actin in T cells is a critical step for immunological synapse formation, and lamin-A interacted with the linker of nucleoskeleton and cytoskeleton (LINC) complex to promote F-actin polymerization. We also showed that lamin-A expression accelerated TCR clustering and led to enhanced downstream signaling, including extracellular signal-regulated kinase 1/2 (ERK1/2) signaling, as well as increased target gene expression. Pharmacological inhibition of the ERK pathway reduced lamin-A-dependent T cell activation. Moreover, mice lacking lamin-A in immune cells exhibited impaired T cell responses in vivo. These findings underscore the importance of A-type lamins for TCR activation and identify lamin-A as a previously unappreciated regulator of the immune response.

  15. Nuclear winds and the narrow-line emission from active galaxies

    NASA Technical Reports Server (NTRS)

    Smith, Steven J.

    1993-01-01

    This paper discusses active galaxy narrow-line emission in the context of a supersonic wind generated by the active nucleus, a nuclear wind. Wind acceleration can naturally produce the inferred narrow-line cloud velocities as well as those of the broad-line and broad absorption line clouds with only a weak dependence on the parameters of the active nucleus. The primary obstacle to wind acceleration of emission-line clouds is the destructive effect of instabilities; however, the stability and structure of wind-confined clouds remain areas of future research. Observations, particularly of the Seyfert galaxy NGC 1068, are interpreted in the context of nuclear winds.

  16. Natural compounds regulate energy metabolism by the modulating the activity of lipid-sensing nuclear receptors.

    PubMed

    Goto, Tsuyoshi; Kim, Young-Il; Takahashi, Nobuyuki; Kawada, Teruo

    2013-01-01

    Obesity causes excess fat accumulation in various tissues, most notoriously in the adipose tissue, along with other insulin-responsive organs such as skeletal muscle and the liver, which predisposes an individual to the development of metabolic abnormalities. The molecular mechanisms underlying obesity-induced metabolic abnormalities have not been completely elucidated; however, in recent years, the search for therapies to prevent the development of obesity and obesity-associated metabolic disorders has increased. It is known that several nuclear receptors, when activated by specific ligands, regulate carbohydrate and lipid metabolism at the transcriptional level. The expression of lipid metabolism-related enzymes is directly regulated by the activity of various nuclear receptors via their interaction with specific response elements in promoters of those genes. Many natural compounds act as ligands of nuclear receptors and regulate carbohydrate and lipid metabolism by regulating the activities of these nuclear receptors. In this review, we describe our current knowledge of obesity, the role of lipid-sensing nuclear receptors in energy metabolism, and several examples of food factors that act as agonists or antagonists of nuclear receptors, which may be useful for the management of obesity and the accompanying energy metabolism abnormalities.

  17. Toxic PRn poly-dipeptides encoded by the C9orf72 repeat expansion block nuclear import and export

    PubMed Central

    Shi, Kevin Y.; Mori, Eiichiro; Nizami, Zehra F.; Lin, Yi; Kato, Masato; Xiang, Siheng; Wu, Leeju C.; Ding, Ming; Yu, Yonghao; Gall, Joseph G.; McKnight, Steven L.

    2017-01-01

    The toxic proline:arginine (PRn) poly-dipeptide encoded by the (GGGGCC)n repeat expansion in the C9orf72 form of heritable amyotrophic lateral sclerosis (ALS) binds to the central channel of the nuclear pore and inhibits the movement of macromolecules into and out of the nucleus. The PRn poly-dipeptide binds to polymeric forms of the phenylalanine:glycine (FG) repeat domain, which is shared by several proteins of the nuclear pore complex, including those in the central channel. A method of chemical footprinting was used to characterize labile, cross-β polymers formed from the FG domain of the Nup54 protein. Mutations within the footprinted region of Nup54 polymers blocked both polymerization and binding by the PRn poly-dipeptide. The aliphatic alcohol 1,6-hexanediol melted FG domain polymers in vitro and reversed PRn-mediated enhancement of the nuclear pore permeability barrier. These data suggest that toxicity of the PRn poly-dipeptide results in part from its ability to lock the FG repeats of nuclear pore proteins in the polymerized state. Our study offers a mechanistic interpretation of PRn poly-dipeptide toxicity in the context of a prominent form of ALS. PMID:28069952

  18. VRK3-mediated nuclear localization of HSP70 prevents glutamate excitotoxicity-induced apoptosis and Aβ accumulation via enhancement of ERK phosphatase VHR activity

    PubMed Central

    Song, Haengjin; Kim, Wanil; Kim, Sung-Hoon; Kim, Kyong-Tai

    2016-01-01

    Most of neurodegenerative disorders are associated with protein aggregation. Glutamate-induced excitotoxicity and persistent extracellular signal-regulated kinase (ERK) activation are also implicated in neurodegenerative diseases. Here, we found that vaccinia-related kinase 3 (VRK3) facilitates nuclear localization of glutamate-induced heat shock protein 70 (HSP70). Nuclear HSP70 leads to enhancement of vaccinia H1-related phosphatase (VHR) activity via protein-protein interaction rather than its molecular chaperone activity, thereby suppressing excessive ERK activation. Moreover, glutamate-induced ERK activation stimulates the expression of HSP70 and VRK3 at the transcriptional level. Downregulation of either VRK3 or HSP70 rendered cells vulnerable to glutamate-induced apoptosis. Overexpression of HSP70 fused to a nuclear localization signal attenuated apoptosis more than HSP70 alone. The importance of nuclear localization of HSP70 in the negative regulation of glutamate-induced ERK activation was further confirmed in VRK3-deficient neurons. Importantly, we showed a positive correlation between levels of VRK3 and HSP70 in the progression of Alzheimer’s and Parkinson’s diseases in humans, and neurons with HSP70 nuclear localization exhibited less Aβ accumulation in brains from patients with Alzheimer’s disease. Therefore, HSP70 and VRK3 could potentially serve as diagnostic and therapeutic targets in neurodegenerative diseases. PMID:27941812

  19. Nuclear physics frontier at RCNP, Osaka University

    SciTech Connect

    Ong, H. J.

    2014-03-05

    Cyclotron accelerator facility and research activities at Research Center for Nuclear Physics (RCNP), Osaka University, are presented. A special focus is given on several topics in nuclear physics where interesting and important experiment results relevant to the nuclear structure as well as the nuclear astrophysics have been reported.

  20. Melphalan-induced apoptosis of EBV-transformed B cells through upregulation of TAp73 and XAF1 and nuclear import of XPA.

    PubMed

    Park, Ga Bin; Kim, Yeong-Seok; Kim, Daejin; Kim, Seonghan; Lee, Hyun-Kyung; Cho, Dae-Ho; Lee, Wang Jae; Hur, Dae Young

    2013-12-15

    Melphalan (Mel) is widely used to treat patients with hematologic cancer, including multiple myeloma, but its mechanism of action in EBV-transformed B cells is poorly described. In this study, we demonstrate a novel mechanism by which transcriptionally active p73 (TAp73) induces translocation of X-linked inhibitor of apoptosis protein-associated factor 1 (XAF1) and xeroderma pigmentosum group A (XPA) during apoptosis caused by Mel treatment. We observed that Mel induced significant generation of reactive oxygen species (ROS) and subsequent apoptosis, as well as an early phosphorylation of p38 MAPK that preceded expression of the mitochondria membrane potential disruption-related molecules and the cleavage of caspases. In particular, Mel led to upregulation of TAp73, XAF1, and Puma and induced XPA nuclear import and translocation of Bax into mitochondria. Mel-induced apoptosis was inhibited by pretreatment with the ROS scavenger 4-amino-2,4-pyrrolidine-dicarboxylic acid (APDC) and the p38 MAPK inhibitor SB203580. We supposed that ROS generation might be the first event in Mel-induced apoptosis, because APDC blocked the increase in ROS, p38 MAPK, and TAp73, but SB203580 did not block ROS generation. Moreover, Mel elicited activation of ATR, and APDC inhibited phosphorylation of ATR but not SB203580. APDC and SB203580 completely blocked XPA and Bax translocation. We conclude that Mel promotes TAp73-mediated XAF1 and Puma expression via ROS generation and ATR/p38 MAPK pathway activation, thereby triggering apoptosis. Our results provide evidence of a novel alternate regulatory mechanism of TAp73 and reveal that Mel may be a therapeutic drug for curing EBV-related malignancies.

  1. Classical Nuclear Hormone Receptor Activity as a Mediator of Complex Concentration Response Relationships for Endocrine Active Compounds

    PubMed Central

    Cookman, Clifford J.; Belcher, Scott M.

    2014-01-01

    Nonmonotonic concentration response relationships are frequently observed for endocrine active ligands that act via nuclear receptors. The curve of best fit for nonmonotonic concentration response relationships are often inverted U-shaped with effects at intermediate concentrations that are different from effects at higher or lower concentrations. Cytotoxicity is a major mode of action responsible for inverted U-shaped concentration response relationships. However, evidence suggests that ligand selectivity, activation of multiple molecular targets, concerted regulation of multiple opposing endpoints, and multiple ligand binding sites within nuclear receptors also contribute to nonmonotonic concentration response relationships of endocrine active ligands. This review reports the current understanding of mechanisms involved in classical nuclear receptor mediated nonmonotonic concentration response relationships with a focus on studies published between 2012 and 2014. PMID:25299165

  2. Common toad Rhinella arenarum (Hensel, 1867) and its importance in assessing environmental health: test of micronuclei and nuclear abnormalities in erythrocytes.

    PubMed

    Pollo, Favio E; Bionda, Clarisa L; Salinas, Zulma A; Salas, Nancy E; Martino, Adolfo L

    2015-09-01

    Anthropogenic activities may generate significant changes in the integrity of aquatic ecosystems, so long-term monitoring of populations that inhabit them is crucial. Counting micronucleated erythrocytes (MN) and erythrocytic nuclear abnormalities (ENA) in peripheral blood is a widely used method for detecting chromosomal damage due to chemical agents in the water. We analyzed MN and ENA frequency in blood obtained from the common toad Rhinella arenarum populations in sites with different degrees of environmental degradation. The results of this study indicate that there is an association between the frequency of micronuclei and nuclear abnormalities and the degree of environmental alteration recorded for the sites studied.

  3. Development of nuclear analysis capabilities for DOE waste management activities

    SciTech Connect

    Parks, C.V.; DeHart, M.D.; Broadhead, B.L.; Hopper, C.M.; Petrie, L.M.

    1998-05-01

    The objective of this project is to develop and demonstrate prototypic analysis capabilities that can be used by the nuclear safety analysis practitioners to: (1) demonstrate a more thorough understanding of the underlying physics phenomena that can lead to improved reliability and defensibility of safety evaluations; and (2) optimize operations related to the handling, storage, transportation, and disposal of fissile material and DOE spent fuel. To address these problems, the project will investigate the implementation of sensitivity and uncertainty methods within existing Monte Carlo codes used for criticality safety analyses, as well as within a new deterministic code that allows specification of arbitrary grids to accurately model the geometry details required in a criticality safety analysis. This capability can facilitate improved estimations of the required subcritical margin and potentially enable the use of a broader range of experiments in the validation process. The new arbitrary-grid radiation transport code will also enable detailed geometric modeling valuable for improved accuracy in application to a myriad of other problems related to waste characterization. Application to these problems will also be explored.

  4. The enhancement of nuclear receptor transcriptional activation by a mouse actin-binding protein, alpha actinin 2.

    PubMed

    Huang, S M; Huang, C J; Wang, W M; Kang, J C; Hsu, W C

    2004-04-01

    The p160 coactivators, steroid receptor coactivator 1, glucocorticoid receptor interacting protein 1 (GRIP1) and the activator of thyroid and retinoic acid receptor, have two activation domains, AD1 and AD2, which transmit the activation signal from the DNA-bound nuclear receptor to the chromatin and/or transcription machinery. In screening for mammalian proteins that bind the AD2 of GRIP1, we identified a mouse actin-binding protein, alpha actinin 2 (mACTN2). mACTN2 was expressed in the heart, skeletal muscle, lung, brain and testis, but there was no expression in the spleen, liver or kidney. Interestingly, the expression level of mACTN2 in the developing embryo depended on the embryonic stage. We further demonstrated that mACTN2 could enhance two transactivation activities of GRIP1, which in turn could enhance the homodimerization of mACTN2. Importantly, mACTN2 not only served as a primary coactivator for androgen receptor, estrogen receptor and thyroid receptor activities, but also acted synergistically with GRIP1 to enhance these nuclear receptor (NR) functions. However, the NR binding motif, LXXLL, conserved in mACTN2 and other actinin family proteins, might be a dispensable domain for its coactivator roles in NRs. These findings suggested that mACTN2 might play an important role in GRIP1-induced NR coactivator functions.

  5. MST1 activation by curcumin mediates JNK activation, Foxo3a nuclear translocation and apoptosis in melanoma cells

    SciTech Connect

    Yu, Teng; Ji, Jiang; Guo, Yong-li

    2013-11-08

    Highlights: •Curcumin activates MST1 in melanoma cells. •MST1 mediates curcumin-induced apoptosis of melanoma cells. •ROS production is involved in curcumin-induced MST1 activation. •MST1 mediates curcumin-induced JNK activation in melanoma cells. •MST1 mediates curcumin-induced Foxo3a nuclear translocation and Bim expression. -- Abstract: Different groups including ours have shown that curcumin induces melanoma cell apoptosis, here we focused the role of mammalian Sterile 20-like kinase 1 (MST1) in it. We observed that curcumin activated MST1-dependent apoptosis in cultured melanoma cells. MST1 silencing by RNA interference (RNAi) suppressed curcumin-induced cell apoptosis, while MST1 over-expressing increased curcumin sensitivity. Meanwhile, curcumin induced reactive oxygen species (ROS) production in melanoma cells, and the ROS scavenger, N-acetyl-cysteine (NAC), almost blocked MST1 activation to suggest that ROS might be required for MST1 activation by curcumin. c-Jun N-terminal protein kinase (JNK) activation by curcumin was dependent on MST1, since MST1 inhibition by RNAi or NAC largely inhibited curcumin-induced JNK activation. Further, curcumin induced Foxo3 nuclear translocation and Bim-1 (Foxo3 target gene) expression in melanoma cells, such an effect by curcumin was inhibited by MST1 RNAi. In conclusion, we suggested that MST1 activation by curcumin mediates JNK activation, Foxo3a nuclear translocation and apoptosis in melanoma cells.

  6. [Activities of administered radiopharmaceuticals and population dose from nuclear medicine in Czechoslovakia].

    PubMed

    Gushak, V; Rzhichkova, G

    1991-01-01

    The authors assessed by means of questionnaires the activities of radiopharmaceuticals administered in departments of nuclear medicine in Czechoslovakia. The mean activities of individual radiopharmaceuticals are roughly equal as in Great Britain but lower than in the Canadian province of Manitoba. The differences of activities used in different departments are approximately equal in all compared countries. In the Czech Republic the annual collective effective dose equivalent from nuclear medicine was 433 Sv in 1983 and 609 Sv in 1987. The mean effective dose equivalent per examination was 2.23 mSv in 1983 and 2.44 mSv in 1987. The mean effective dose equivalent per inhabitant of the Czech Republic was 0.042 mSv in 1983 and 0.059 mSv in 1987. The radiation dose of the Czech population from nuclear medicine amounts approximately to one tenth of the load from radiodiagnostics.

  7. Activities of the OECD/NEA Expert Group on Assay Data for Spent Nuclear Fuel

    SciTech Connect

    Gauld, Ian C; Rugama, Yolanda

    2009-01-01

    Management of spent nuclear fuel is a key issue for many NEA member countries. In nuclear criticality safety, the decision of many countries to advance burnup credit as part of their licensing strategy has heightened recent interest in experimental data needed to validate computer codes used in burnup credit calculations. This paper discusses recent activities of an Expert Group on assay data, formed under the OECD/NEA/NSC/WPNCS (Working Party on Nuclear Criticality Safety) to help coordinate isotopic assay data activities and facilitate international collaboration between NEA member countries developing or implementing burnup credit methodologies. Recent activities of the Expert Group are described, focusing on the planned expansion of the Spent Fuel Isotopic Composition Database (SFCOMPO), and preparation of a state-of-the-art report on assay data that includes sections on recommended radiochemical analysis methods, techniques, and lessons learned from previous experiments.

  8. Radioisotopes "economy of promises": on the limits of biomedicine in public legitimization of nuclear activities.

    PubMed

    Boudia, Soraya

    2009-01-01

    This paper aims to examine the rise and the fall of biomedicine in the public legitimization of the development of nuclear energy. Until the late 1950s, biological and medical applications of radioisotopes were presented as the most important successes of the peaceful uses of atomic energy. I will argue that despite the major financial investment, the development of the uses of radioisotopes and their important impact on biology and clinical practices, the assessment of medical uses remained relatively limited. As consequence, the place of biomedicine in the public legitimization of financial investment and civilian uses of nuclear energy began to decline from the late 1950s.

  9. Drosophila SAF-B Links the Nuclear Matrix, Chromosomes, and Transcriptional Activity

    PubMed Central

    Alfonso-Parra, Catalina; Maggert, Keith A.

    2010-01-01

    Induction of gene expression is correlated with alterations in nuclear organization, including proximity to other active genes, to the nuclear cortex, and to cytologically distinct domains of the nucleus. Chromosomes are tethered to the insoluble nuclear scaffold/matrix through interaction with Scaffold/Matrix Attachment Region (SAR/MAR) binding proteins. Identification and characterization of proteins involved in establishing or maintaining chromosome-scaffold interactions is necessary to understand how the nucleus is organized and how dynamic changes in attachment are correlated with alterations in gene expression. We identified and characterized one such scaffold attachment factor, a Drosophila homolog of mammalian SAF-B. The large nuclei and chromosomes of Drosophila have allowed us to show that SAF-B inhabits distinct subnuclear compartments, forms weblike continua in nuclei of salivary glands, and interacts with discrete chromosomal loci in interphase nuclei. These interactions appear mediated either by DNA-protein interactions, or through RNA-protein interactions that can be altered during changes in gene expression programs. Extraction of soluble nuclear proteins and DNA leaves SAF-B intact, showing that this scaffold/matrix-attachment protein is a durable component of the nuclear matrix. Together, we have shown that SAF-B links the nuclear scaffold, chromosomes, and transcriptional activity. PMID:20422039

  10. NASA safety program activities in support of the Space Exploration Initiatives Nuclear Propulsion program

    NASA Technical Reports Server (NTRS)

    Sawyer, J. C., Jr.

    1993-01-01

    The activities of the joint NASA/DOE/DOD Nuclear Propulsion Program Technical Panels have been used as the basis for the current development of safety policies and requirements for the Space Exploration Initiatives (SEI) Nuclear Propulsion Technology development program. The Safety Division of the NASA Office of Safety and Mission Quality has initiated efforts to develop policies for the safe use of nuclear propulsion in space through involvement in the joint agency Nuclear Safety Policy Working Group (NSPWG), encouraged expansion of the initial policy development into proposed programmatic requirements, and suggested further expansion into the overall risk assessment and risk management process for the NASA Exploration Program. Similar efforts are underway within the Department of Energy to ensure the safe development and testing of nuclear propulsion systems on Earth. This paper describes the NASA safety policy related to requirements for the design of systems that may operate where Earth re-entry is a possibility. The expected plan of action is to support and oversee activities related to the technology development of nuclear propulsion in space, and support the overall safety and risk management program being developed for the NASA Exploration Program.

  11. Characterization of the subcellular localization and nuclear import molecular mechanisms of herpes simplex virus 1 UL2.

    PubMed

    Cai, Mingsheng; Huang, Zebin; Liao, Zongmin; Chen, Tao; Wang, Ping; Jiang, Si; Chen, Daixiong; Peng, Tao; Bian, Yun; Hong, Gengde; Yang, Hang; Zeng, Zhancheng; Li, Xiaowei; Li, Meili

    2017-04-01

    As a crucial protein, the herpes simplex virus 1 (HSV-1) UL2 protein has been shown to take part in various stages of viral infection, nonetheless, its exact subcellular localization and transport molecular determinants are not well known thus far. In the present study, by using live cells fluorescent microscopy assay, UL2 tagged with enhanced yellow fluorescent protein was transiently expressed in live cells and showed a completely nuclear accumulation without the presence of other HSV-1 proteins. Moreover, the nuclear transport of UL2 was characterized to be assisted by multiple transport pathways through Ran-, importin α1-, α5-, α7-, β1- and transportin-1 cellular transport receptors. Consequently, these results will improve understanding of UL2-mediated biological functions in HSV-1 infection cycles.

  12. Sesamin attenuates allergic airway inflammation through the suppression of nuclear factor-kappa B activation.

    PubMed

    Li, Liangchang; Piao, Hongmei; Zheng, Mingyu; Jin, Zhewu; Zhao, Liguang; Yan, Guanghai

    2016-12-01

    The aim of the present study is to determine the role of sesamin, the most abundant lignan in sesame seed oil, on the regulation of allergic airway inflammation in a murine asthma model. A BALB/c mouse model with allergic asthma was used to evaluate the effects of sesamin on nuclear factor-kappa B (NF-κB) activation. An enzyme-linked immunosorbent assay was used to determine protein expression in bronchoalveolar lavage (BAL) fluids. Hematoxylin and eosin staining was performed to examine histological changes. Moreover, western blot analysis was used to detect the expression of proteins in tissues. Prior to administering sesamin, the mice developed the following pathophysiological features of asthma: An increase in the number of inflammatory cells, increased levels of interleukin (IL)-4, IL-5 and IL-13, decreased levels of interferon-γ in BAL fluids and lung tissues, increased immunoglobulin E (IgE) levels in the serum and an increased activation of NF-κB in lung tissues. Following treatment with sesamin, the mice had evidently reduced peribronchiolar inflammation and airway inflammatory cell recruitment, inhibited production of several cytokines in BAL fluids and lung tissues, and decreased IgE levels. Following inhalation of ovalbumin, the administration of sesamin also inhibited the activation of NF-κB. In addition, sesamin administration reduced the phosphorylation of p38 mitogen-activated protein kinases (MAPKs). The present study demonstrates that sesamin decreases the activation of NF-κB in order to attenuate allergic airway inflammation in a murine model of asthma, possibly via the regulation of phosphorylation of p38 MAPK. These observations provide an important molecular mechanism for the potential use of sesamin in preventing and/or treating asthma, as well as other airway inflammatory disorders.

  13. Sesamin attenuates allergic airway inflammation through the suppression of nuclear factor-kappa B activation

    PubMed Central

    Li, Liangchang; Piao, Hongmei; Zheng, Mingyu; Jin, Zhewu; Zhao, Liguang; Yan, Guanghai

    2016-01-01

    The aim of the present study is to determine the role of sesamin, the most abundant lignan in sesame seed oil, on the regulation of allergic airway inflammation in a murine asthma model. A BALB/c mouse model with allergic asthma was used to evaluate the effects of sesamin on nuclear factor-kappa B (NF-κB) activation. An enzyme-linked immunosorbent assay was used to determine protein expression in bronchoalveolar lavage (BAL) fluids. Hematoxylin and eosin staining was performed to examine histological changes. Moreover, western blot analysis was used to detect the expression of proteins in tissues. Prior to administering sesamin, the mice developed the following pathophysiological features of asthma: An increase in the number of inflammatory cells, increased levels of interleukin (IL)-4, IL-5 and IL-13, decreased levels of interferon-γ in BAL fluids and lung tissues, increased immunoglobulin E (IgE) levels in the serum and an increased activation of NF-κB in lung tissues. Following treatment with sesamin, the mice had evidently reduced peribronchiolar inflammation and airway inflammatory cell recruitment, inhibited production of several cytokines in BAL fluids and lung tissues, and decreased IgE levels. Following inhalation of ovalbumin, the administration of sesamin also inhibited the activation of NF-κB. In addition, sesamin administration reduced the phosphorylation of p38 mitogen-activated protein kinases (MAPKs). The present study demonstrates that sesamin decreases the activation of NF-κB in order to attenuate allergic airway inflammation in a murine model of asthma, possibly via the regulation of phosphorylation of p38 MAPK. These observations provide an important molecular mechanism for the potential use of sesamin in preventing and/or treating asthma, as well as other airway inflammatory disorders. PMID:28105144

  14. Physical Activity in the Heat: Important Considerations to Keep Your Students Safe

    ERIC Educational Resources Information Center

    Roetert, E. Paul; Richardson, Cheryl L.; Bergeron, Michael F.

    2012-01-01

    Although July and August tend to be the warmest months of the year, the months leading up to summer as well as the months just following summer can also be quite warm or even very hot. In this article, the authors share some important information to help prepare physical educators for overseeing activities in the heat and, just as importantly, to…

  15. Active fault and water loading are important factors in triggering earthquake activity around Aswan Lake

    NASA Astrophysics Data System (ADS)

    Kebeasy, R. M.; Gharib, A. A.

    Aswan Lake started impounding in 1964 and reached the highest water level so far in 1978 with a capacity of 133.8 km 3, thus forming the second largest man-made lake in the world. An earthquake of magnitude 5.3 (Ms) took place on 14 November 1981 along the most active part of the E-W Kalabsha fault beneath the Kalabsha bay (the largest bay of the lake). This earthquake was followed by a tremendous number of smaller events that continue till now. A radio-telemetry network of 13 seismic short period stations and a piezometer network of six wells were established around the northern part of the lake. Epicenters were found to cluster around active faults near the lake. The space-time distribution and the relation of the seismicity with the lake water level fluctuations were studied. Six years after flooding the eastern segment of the Kalabsha fault, strong seismicity began following the main shock of 14 November 1981. It occurred four days after the reservoir had reached its seasonal max level. The effect of the North African drought (1982 to present) is clearly seen in the reservoir water level. As it decreased and left the most active fault segments uncovered, the activity (Gebel Marawa area) decreased sharply. Also, the shallow activity was found to be more sensitive to rapid discharging than to the filling. This study indicates that geology, topography, lineations in seismicity, offsets in the faults, changes in fault trends and focal mechanisms are closely related. No relation was found between earthquake activity and both-ground water table fluctuations and water temperatures measured in wells located around the Kalabsha area.

  16. Differential influence of instruments in nuclear core activity evaluation by data assimilation

    NASA Astrophysics Data System (ADS)

    Bouriquet, Bertrand; Argaud, Jean-Philippe; Erhard, Patrick; Massart, Sébastien; Ponçot, Angélique; Ricci, Sophie; Thual, Olivier

    2011-01-01

    The global neutronic activity fields of a nuclear core can be reconstructed using data assimilation. Indeed, data assimilation allows to combine both measurements from instruments and information from a model, to evaluate the best possible neutronic activity within the core. We present and apply a specific procedure which evaluates the influence of measures by adding or removing instruments in a given measurement network (possibly empty). The study of various network configurations for the instruments in the nuclear core establishes that the influence of the instruments depends both on the independent instrumentation location and on the chosen network.

  17. Alanine-Scanning Mutational Analysis of Durancin GL Reveals Residues Important for Its Antimicrobial Activity.

    PubMed

    Ju, Xingrong; Chen, Xinquan; Du, Lihui; Wu, Xueyou; Liu, Fang; Yuan, Jian

    2015-07-22

    Durancin GL is a novel class IIa bacteriocin with 43 residues produced by Enterococcus durans 41D. This bacteriocin demonstrates narrow inhibition spectrum and potent antimicrobial activity against several Listeria monocytogenes strains, including nisin-resistant L. monocytogenes NR30. A systematic alanine-scanning mutational analysis with site-directed mutagenesis was performed to analyze durancin GL residues important for antimicrobial activity and specificity. Results showed that three mutations lost their antimicrobial activity, ten mutations demonstrated a decreased effect on the activity, and seven mutations exhibited relatively high activity. With regard to inhibitory spectrum, four mutants demonstrated a narrower antimicrobial spectrum than wild-type durancin GL. Another four mutants displayed a broader target cell spectrum and increased potency relative to wild-type durancin GL. These findings broaden our understanding of durancin GL residues important for its antimicrobial activity and contribute to future rational design of variants with increased potency.

  18. Tritium activities in Canada supporting CANDU{sup R} nuclear power reactors

    SciTech Connect

    Miller, J. M.

    2008-07-15

    An overview of the various Canadian tritium research and operational activities supporting the development, refurbishment and operation of CANDU{sup R} nuclear power reactors is presented. These activities encompass tritium health and safety, tritium in the environment, tritium interaction with materials, and tritium processing, and relate to both supporting R and D advances as well as operational best practices. The collective results of these activities contribute to our goals of improving worker and public safety, and operational efficiency. (authors)

  19. Obox4-silencing-activated STAT3 and MPF/MAPK signaling accelerate nuclear membrane breakdown in mouse oocytes.

    PubMed

    Lee, Hyun-Seo; Kim, Kyeoung-Hwa; Kim, Eun-Young; Lee, Su-Yeon; Ko, Jung-Jae; Lee, Kyung-Ah

    2016-04-01

    Mouse oocytes begin to mature in vitro once liberated from ovarian follicles. Previously, we showed that oocyte-specific homeobox 4 (Obox4) is critical for maintaining the intact nuclear membrane of the germinal vesicle (GV) in oocytes and for completing meiosis at the metaphase I-II (MI-MII) transition. This study further examines the molecular mechanisms of OBOX4 in regulating GV nuclear membrane breakdown. Maturation-promoting factor (MPF) and MAPK are normally inactive in GV stage oocytes but were activated prematurely in arrested GV stage oocytes by 3-isobutyl-1-metyl-xanthine (IBMX) in vitro after Obox4 RNA interference (RNAi). Furthermore, signal transducer and activator of transcription 3 (STAT3) was significantly activated by Obox4 RNAi. We confirmed that this Obox4 RNAi-induced premature STAT3 and MPF/MAPK activation at the GV stage provoked subsequent GV breakdown (GVBD) despite the opposing force of high cAMP in the IBMX-supplemented medium to maintain intact GV. When cumulus-oocyte complexes were exposed to interferon α (IFNA), a STAT3 activator, oocytes matured and cumulus cells expanded to resume nuclear maturation in IBMX-supplemented medium, suggesting that STAT3 activation is sufficient for stimulating the continuation of meiosis. Using Stattic, a specific STAT3 inhibitor, we confirmed that GVBD involves STAT3 activation in Obox4-silenced oocytes. Based on these findings, we concluded that i) Obox4 is an important upstream regulator of MPF/MAPK and STAT3 signaling, and ii) Obox4 is a key regulator of the GV arrest mechanism in oocytes.

  20. Nuclear Import of Adenovirus DNA Involves Direct Interaction of Hexon with an N-Terminal Domain of the Nucleoporin Nup214

    PubMed Central

    Ragues, Jessica; Guan, Tinglu; Bégu, Dominique; Wodrich, Harald; Kann, Michael; Nemerow, Glen R.; Gerace, Larry

    2014-01-01

    ABSTRACT In this study, we characterized the molecular basis for binding of adenovirus (AdV) to the cytoplasmic face of the nuclear pore complex (NPC), a key step during delivery of the viral genome into the nucleus. We used RNA interference (RNAi) to deplete cells of either Nup214 or Nup358, the two major Phe-Gly (FG) repeat nucleoporins localized on the cytoplasmic side of the NPC, and evaluated the impact on hexon binding and AdV infection. The accumulation of purified hexon trimers or partially disassembled AdV at the nuclear envelope (NE) was observed in digitonin-permeabilized cells in the absence of cytosolic factors. Both in vitro hexon binding and in vivo nuclear import of the AdV genome were strongly reduced in Nup214-depleted cells but still occurred in Nup358-depleted cells, suggesting that Nup214 is a major binding site of AdV during infection. The expression of an NPC-targeted N-terminal domain of Nup214 in Nup214-depleted cells restored the binding of hexon at the NE and the nuclear import of protein VII (pVII), indicating that this region is sufficient to allow AdV binding. We further narrowed the binding site to a 137-amino-acid segment in the N-terminal domain of Nup214. Together, our results have identified a specific region within the N terminus of Nup214 that acts as a direct NPC binding site for AdV. IMPORTANCE AdVs, which have the largest genome of nonenveloped DNA viruses, are being extensively explored for use in gene therapy, especially in alternative treatments for cancers that are refractory to traditional therapies. In this study, we characterized the molecular basis for binding of AdV to the cytoplasmic face of the NPC, a key step for delivery of the viral genome into the nucleus. Our data indicate that a 137-amino-acid region of the nucleoporin Nup214 is a binding site for the major AdV capsid protein, hexon, and that this interaction is required for viral DNA import. These findings provide additional insight on how AdV exploits the

  1. Activated nuclear metabotropic glutamate receptor mGlu5 couples to nuclear Gq/11 proteins to generate inositol 1,4,5-trisphosphate-mediated nuclear Ca2+ release.

    PubMed

    Kumar, Vikas; Jong, Yuh-Jiin I; O'Malley, Karen L

    2008-05-16

    Recently we have shown that the metabotropic glutamate 5 (mGlu5) receptor can be expressed on nuclear membranes of heterologous cells or endogenously on striatal neurons where it can mediate nuclear Ca2+ changes. Here, pharmacological, optical, and genetic techniques were used to show that upon activation, nuclear mGlu5 receptors generate nuclear inositol 1,4,5-trisphosphate (IP3) in situ. Specifically, expression of an mGlu5 F767S mutant in HEK293 cells that blocks Gq/11 coupling or introduction of a dominant negative Galphaq construct in striatal neurons prevented nuclear Ca2+ changes following receptor activation. These data indicate that nuclear mGlu5 receptors couple to Gq/11 to mobilize nuclear Ca2+. Nuclear mGlu5-mediated Ca2+ responses could also be blocked by the phospholipase C (PLC) inhibitor, U73122, the phosphatidylinositol (PI) PLC inhibitor 1-O-octadecyl-2-O-methyl-sn-glycero-3-phosphorylcholine (ET-18-OCH3), or by using small interfering RNA targeted against PLCbeta1 demonstrating that PI-PLC is involved. Direct assessment of inositol phosphate production using a PIP2/IP3 "biosensor" revealed for the first time that IP3 can be generated in the nucleus following activation of nuclear mGlu5 receptors. Finally, both IP3 and ryanodine receptor blockers prevented nuclear mGlu5-mediated increases in intranuclear Ca2+. Collectively, this study shows that like plasma membrane receptors, activated nuclear mGlu5 receptors couple to Gq/11 and PLC to generate IP3-mediated release of Ca2+ from Ca2+-release channels in the nucleus. Thus the nucleus can function as an autonomous organelle independent of signals originating in the cytoplasm, and nuclear mGlu5 receptors play a dynamic role in mobilizing Ca2+ in a specific, localized fashion.

  2. Chronic exposure to sub-lethal beta-amyloid (Abeta) inhibits the import of nuclear-encoded proteins to mitochondria in differentiated PC12 cells.

    PubMed

    Sirk, Daniel; Zhu, Ziping; Wadia, Jehangir S; Shulyakova, Natalya; Phan, Nam; Fong, Jamie; Mills, Linda R

    2007-12-01

    Studies on amyloid beta (Abeta|), the peptide thought to play a crucial role in the pathogenesis of Alzheimer's disease, have implicated mitochondria in Abeta-mediated neurotoxicity. We used differentiated PC12 cells stably transfected with an inducible green fluorescent protein (GFP) fusion protein containing an N'-terminal mitochondrial targeting sequence (mtGFP), to examine the effects of sub-lethal Abeta on the import of nuclear-encoded proteins to mitochondria. Exposure to sub-lethal Abeta(25-35) (10 mumol/L) for 48 h inhibited mtGFP import to mitochondria; average rates decreased by 20 +/- 4%. Concomitant with the decline in mtGFP, cytoplasmic mtGFP increased significantly while mtGFP expression and intramitochondrial mtGFP turnover were unchanged. Sub-lethal Abeta(1-42) inhibited mtGFP import and increased cytoplasmic mtGFP but only after 96 h. The import of two endogenous nuclear-encoded mitochondrial proteins, mortalin/mtHsp70 and Tom20 also declined. Prior to the decline in import, mitochondrial membrane potential (mmp), and reactive oxygen species levels were unchanged in Abeta-treated cells versus reverse phase controls. Sustained periods of decreased import were associated with decreased mmp, increased reactive oxygen species, increased vulnerability to oxygen-glucose deprivation and altered mitochondrial morphology. These findings suggest that an Abeta-mediated inhibition of mitochondrial protein import, and the consequent mitochondrial impairment, may contribute to Alzheimer's disease.

  3. The importance of input variables to a neural network fault-diagnostic system for nuclear power plants

    SciTech Connect

    Lanc, T.L.

    1992-01-01

    This thesis explores safety enhancement for nuclear power plants. Emergency response systems currently in use depend mainly on automatic systems engaging when certain parameters go beyond a pre-specified safety limit. Often times the operator has little or no opportunity to react since a fast scram signal shuts down the reactor smoothly and efficiently. These accidents are of interest to technical support personnel since examining the conditions that gave rise to these situations help determine causality. In many other cases an automated fault-diagnostic advisor would be a valuable tool in assisting the technicians and operators to determine what just happened and why.

  4. The importance of input variables to a neural network fault-diagnostic system for nuclear power plants

    SciTech Connect

    Lanc, T.L.

    1992-12-31

    This thesis explores safety enhancement for nuclear power plants. Emergency response systems currently in use depend mainly on automatic systems engaging when certain parameters go beyond a pre-specified safety limit. Often times the operator has little or no opportunity to react since a fast scram signal shuts down the reactor smoothly and efficiently. These accidents are of interest to technical support personnel since examining the conditions that gave rise to these situations help determine causality. In many other cases an automated fault-diagnostic advisor would be a valuable tool in assisting the technicians and operators to determine what just happened and why.

  5. Structural basis for the nuclear export activity of Importin13

    PubMed Central

    Grünwald, Marlene; Lazzaretti, Daniela; Bono, Fulvia

    2013-01-01

    Importin13 (Imp13) is a bidirectional karyopherin that can mediate both import and export of cargoes. Imp13 recognizes several import cargoes, which include the exon junction complex components Mago-Y14 and the E2 SUMO-conjugating enzyme Ubc9, and one known export cargo, the translation initiation factor 1A (eIF1A). To understand how Imp13 can perform double duty, we determined the 3.6-Å crystal structure of Imp13 in complex with RanGTP and with eIF1A. eIF1A binds at the inner surface of the Imp13 C-terminal arch adjacent and concomitantly to RanGTP illustrating how eIF1A can be exported by Imp13. Moreover, the 3.0-Å structure of Imp13 in its unbound state reveals the existence of an open conformation in the cytoplasm that explains export cargo release and completes the export branch of the Imp13 pathway. Finally, we demonstrate that Imp13 is able to bind and export eIF1A in vivo and that its function is essential. PMID:23435562

  6. Activities on Nuclear Data Measurements at Pohang Neutron Facility

    NASA Astrophysics Data System (ADS)

    Kim, Guinyun

    2009-03-01

    We report the activities of the Pohang Neutron Facility which consists of an electron linear accelerator, a water-cooled Ta target, and a 12-m time-of-flight path. It has been equipped with a four-position sample changer controlled remotely by a CAMAC data acquisition system, which allows simultaneous accumulation of the neutron time of flight spectra from 4 different detectors. It can be possible to measure the neutron total cross-sections in the neutron energy range from 0.1 eV to few hundreds eV by using the neutron time-of-flight method. A 6LiZnS(Ag) glass scintillator was used as a neutron detector. The neutron flight path from the water-cooled Ta target to the neutron detector was 12.1 m. The background level was determined by using notch-filters of Co, In, Ta, and Cd sheets. In order to reduce the gamma rays from bremsstrahlung and those from neutron capture, we employed a neutron-gamma separation system based on their different pulse shapes. The present measurements of several samples (Ta, Mo) are in general agreement with the evaluated data in ENDF/B-VI. We measured the thermal neutron capture cross-sections and the resonance integrals of the 186W(n,γ)187W reaction and the 98Mo(n,γ)99Mo reaction by the activation method using the 197Au(n,γ)198Au monitor reaction as a single comparator. We also report the isomeric yield ratios for the 44 m, gSc isomeric pairs produced from four different photonuclear reactions 45Sc(γ,n)44m,gSc, natTi(γ,xn1p)44m,gSc, natFe(γ,xn5p)52m,gMn, and 103Rh(γ,4n)99m,gRh by using the activation method.

  7. Extensive mutagenesis of a transcriptional activation domain identifies single hydrophobic and acidic amino acids important for activation in vivo.

    PubMed Central

    Sainz, M B; Goff, S A; Chandler, V L

    1997-01-01

    C1 is a transcriptional activator of genes encoding biosynthetic enzymes of the maize anthocyanin pigment pathway. C1 has an amino terminus homologous to Myb DNA-binding domains and an acidic carboxyl terminus that is a transcriptional activation domain in maize and yeast cells. To identify amino acids critical for transcriptional activation, an extensive random mutagenesis of the C1 carboxyl terminus was done. The C1 activation domain is remarkably tolerant of amino acid substitutions, as changes at 34 residues had little or no effect on transcriptional activity. These changes include introduction of helix-incompatible amino acids throughout the C1 activation domain and alteration of most single acidic amino acids, suggesting that a previously postulated amphipathic alpha-helix is not required for activation. Substitutions at two positions revealed amino acids important for transcriptional activation. Replacement of leucine 253 with a proline or glutamine resulted in approximately 10% of wild-type transcriptional activation. Leucine 253 is in a region of C1 in which several hydrophobic residues align with residues important for transcriptional activation by the herpes simplex virus VP16 protein. However, changes at all other hydrophobic residues in C1 indicate that none are critical for C1 transcriptional activation. The other important amino acid in C1 is aspartate 262, as a change to valine resulted in only 24% of wild-type transcriptional activation. Comparison of our C1 results with those from VP16 reveal substantial differences in which amino acids are required for transcriptional activation in vivo by these two acidic activation domains. PMID:8972191

  8. 24th WRSM panel discussion: {open_quotes}The role of research in nuclear regulation: The case of qualified importers{close_quotes}

    SciTech Connect

    Alonso, A.

    1997-01-01

    Scientific knowledge and technological maturity are needed to establish regulatory requirements, what also needs talent and skills. Scientists are rarely interested in regulation and regulators not always are closely connected to scientific research. This has created gaps in regulations, mainly within qualified importers. A qualified importer, in the sense of this presentation, is a country who has acquired nuclear power plants from more technologically advanced exporters but with an increasing participation of its own industry and institutions in the design, construction, component manufacture and assembly of such nuclear power plants and is fully responsible for the operation of the nuclear units and the corresponding fuel cycle. These countries have also a long standing and independent nuclear regulatory organization and the corresponding technical body. Spain is a qualified importer. In the case of Spain, it originally adopted the codes and regulations of the UE which it was a member of, as well as the codes and standards of the country from which their reactors were produced. Since Spain added KWU plants they even went to German regulations. Plant operation rested with Spain. As problems began to appear in operating plants, local research projects were funded to study the problems, aimed at immediate solutions, but also providing training for local personnel in addition to information of use for regulators. Spain has participated in many joint research projects, which have trained people, and given confidence to Spaniards involved in science and regulations. Qualified importers, like Spain, participate in research, even though it may not translate itself into regulation. Such participation will always serve to give self-confidence and independence to regulators and licensees, to better understand the adopted regulations of the most advanced countries and to solve specific problems.

  9. Syntaxin 6-mediated Golgi translocation plays an important role in nuclear functions of EGFR through microtubule-dependent trafficking

    PubMed Central

    Du, Y; Shen, J; Hsu, JL; Han, Z; Hsu, M-C; Yang, C-C; Kuo, H-P; Wang, Y-N; Yamaguchi, H; Miller, SA; Hung, M-C

    2013-01-01

    Receptor tyrosine kinases (RTKs) are cell surface receptors that initiate signal cascades in response to ligand stimulation. Abnormal expression and dysregulated intracellular trafficking of RTKs have been shown to be involved in tumorigenesis. Recent evidence shows that these cell surface receptors translocate from cell surface to different cellular compartments, including the Golgi, mitochondria, endoplasmic reticulum (ER) and the nucleus, to regulate physiological and pathological functions. Although some trafficking mechanisms have been resolved, the mechanism of intracellular trafficking from cell surface to the Golgi is not yet completely understood. Here we report a mechanism of Golgi translocation of epidermal growth factor receptor (EGFR) in which EGF-induced EGFR travels to the Golgi via microtubule-dependent movement by interacting with dynein and fuses with the Golgi through syntaxin 6-mediated membrane fusion. We also demonstrate that the microtubule- and syntaxin 6-mediated Golgi translocation of EGFR is necessary for its consequent nuclear translocation and nuclear functions. Thus, together with previous studies, the microtubule- and syntaxin 6-mediated trafficking pathway from cell surface to the Golgi, ER and the nucleus defines a comprehensive trafficking route for EGFR to travel from cell surface to the Golgi and the nucleus. PMID:23376851

  10. Syntaxin 6-mediated Golgi translocation plays an important role in nuclear functions of EGFR through microtubule-dependent trafficking.

    PubMed

    Du, Y; Shen, J; Hsu, J L; Han, Z; Hsu, M-C; Yang, C-C; Kuo, H-P; Wang, Y-N; Yamaguchi, H; Miller, S A; Hung, M-C

    2014-02-06

    Receptor tyrosine kinases (RTKs) are cell surface receptors that initiate signal cascades in response to ligand stimulation. Abnormal expression and dysregulated intracellular trafficking of RTKs have been shown to be involved in tumorigenesis. Recent evidence shows that these cell surface receptors translocate from cell surface to different cellular compartments, including the Golgi, mitochondria, endoplasmic reticulum (ER) and the nucleus, to regulate physiological and pathological functions. Although some trafficking mechanisms have been resolved, the mechanism of intracellular trafficking from cell surface to the Golgi is not yet completely understood. Here we report a mechanism of Golgi translocation of epidermal growth factor receptor (EGFR) in which EGF-induced EGFR travels to the Golgi via microtubule-dependent movement by interacting with dynein and fuses with the Golgi through syntaxin 6-mediated membrane fusion. We also demonstrate that the microtubule- and syntaxin 6-mediated Golgi translocation of EGFR is necessary for its consequent nuclear translocation and nuclear functions. Thus, together with previous studies, the microtubule- and syntaxin 6-mediated trafficking pathway from cell surface to the Golgi, ER and the nucleus defines a comprehensive trafficking route for EGFR to travel from cell surface to the Golgi and the nucleus.

  11. Inhibition of human immunodeficiency virus type 1 (HIV-1) nuclear import via Vpr-Importin {alpha} interactions as a novel HIV-1 therapy

    SciTech Connect

    Suzuki, Tatsunori; Yamamoto, Norio; Nonaka, Mizuho; Hashimoto, Yoshie; Matsuda, Go; Takeshima, Shin-nosuke; Matsuyama, Megumi; Igarashi, Tatsuhiko; Miura, Tomoyuki; Tanaka, Rie; Kato, Shingo; Aida, Yoko

    2009-03-20

    The development of multidrug-resistant viruses compromises the efficacy of anti-human immunodeficiency virus (HIV) therapy and limits treatment options. Therefore, new targets that can be used to develop novel antiviral agents need to be identified. One such target is the interaction between Vpr, one of the accessory gene products of HIV-1 and Importin {alpha}, which is crucial, not only for the nuclear import of Vpr, but also for HIV-1 replication in macrophages. We have identified a potential parent compound, hematoxylin, which suppresses Vpr-Importin {alpha} interaction, thereby inhibiting HIV-1 replication in a Vpr-dependent manner. Analysis by real-time PCR demonstrated that hematoxylin specifically inhibited nuclear import step of pre-integration complex. Thus, hematoxylin is a new anti-HIV-1 inhibitor that targets the nuclear import of HIV-1 via the Vpr-Importin {alpha} interaction, suggesting that a specific inhibitor of the interaction between viral protein and the cellular factor may provide a new strategy for HIV-1 therapy.

  12. Active nondestructive assay of nuclear materials: principles and applications

    SciTech Connect

    Gozani, Tsahi

    1981-01-01

    The purpose of this book is to present, coherently and comprehensively, the wealth of available but scattered information on the principles and applications of active nondestructive analysis (ANDA). Chapters are devoted to the following: background and overview; interactions of neutrons with matter; interactions of ..gamma..-rays with matter; neutron production and sources; ..gamma..-ray production and sources; effects of neutron and ..gamma..-ray transport in bulk media; signatures of neutron- and photon-induced fissions; neutron and photon detection systems and electronics; representative ANDA systems; and instrument analysis, calibration, and measurement control for ANDA. Each chapter has an introductory section describing the relationship of the topic of that chapter to ANDA. Each chapter ends with a section that summarizes the main results and conclusions of the chapter, and a reference list.

  13. Thermodynamics of nuclear transport

    NASA Astrophysics Data System (ADS)

    Wang, Ching-Hao; Mehta, Pankaj; Elbaum, Michael

    Molecular transport across the nuclear envelope is important for eukaryotes for gene expression and signaling. Experimental studies have revealed that nuclear transport is inherently a nonequilibrium process and actively consumes energy. In this work we present a thermodynamics theory of nuclear transport for a major class of nuclear transporters that are mediated by the small GTPase Ran. We identify the molecular elements responsible for powering nuclear transport, which we term the ``Ran battery'' and find that the efficiency of transport, measured by the cargo nuclear localization ratio, is limited by competition between cargo molecules and RanGTP to bind transport receptors, as well as the amount of NTF2 (i.e. RanGDP carrier) available to circulate the energy flow. This picture complements our current understanding of nuclear transport by providing a comprehensive thermodynamics framework to decipher the underlying biochemical machinery. Pm and CHW were supported by a Simons Investigator in the Mathematical Modeling in Living Systems grant (to PM).

  14. The transcriptional activity of hepatocyte nuclear factor 4 alpha is inhibited via phosphorylation by ERK1/2.

    PubMed

    Vető, Borbála; Bojcsuk, Dóra; Bacquet, Caroline; Kiss, Judit; Sipeki, Szabolcs; Martin, Ludovic; Buday, László; Bálint, Bálint L; Arányi, Tamás

    2017-01-01

    Hepatocyte nuclear factor 4 alpha (HNF4α) nuclear receptor is a master regulator of hepatocyte development, nutrient transport and metabolism. HNF4α is regulated both at the transcriptional and post-transcriptional levels by different mechanisms. Several kinases (PKA, PKC, AMPK) were shown to phosphorylate and decrease the activity of HNF4α. Activation of the ERK1/2 signalling pathway, inducing proliferation and survival, inhibits the expression of HNF4α. However, based on our previous results we hypothesized that HNF4α is also regulated at the post-transcriptional level by ERK1/2. Here we show that ERK1/2 is capable of directly phosphorylating HNF4α in vitro at several phosphorylation sites including residues previously shown to be targeted by other kinases, as well. Furthermore, we also demonstrate that phosphorylation of HNF4α leads to a reduced trans-activational capacity of the nuclear receptor in luciferase reporter gene assay. We confirm the functional relevance of these findings by demonstrating with ChIP-qPCR experiments that 30-minute activation of ERK1/2 leads to reduced chromatin binding of HNF4α. Accordingly, we have observed decreasing but not disappearing binding of HNF4α to the target genes. In addition, 24-hour activation of the pathway further decreased HNF4α chromatin binding to specific loci in ChIP-qPCR experiments, which confirms the previous reports on the decreased expression of the HNF4a gene due to ERK1/2 activation. Our data suggest that the ERK1/2 pathway plays an important role in the regulation of HNF4α-dependent hepatic gene expression.

  15. The transcriptional activity of hepatocyte nuclear factor 4 alpha is inhibited via phosphorylation by ERK1/2

    PubMed Central

    Bacquet, Caroline; Kiss, Judit; Sipeki, Szabolcs; Martin, Ludovic; Buday, László; Bálint, Bálint L.; Arányi, Tamás

    2017-01-01

    Hepatocyte nuclear factor 4 alpha (HNF4α) nuclear receptor is a master regulator of hepatocyte development, nutrient transport and metabolism. HNF4α is regulated both at the transcriptional and post-transcriptional levels by different mechanisms. Several kinases (PKA, PKC, AMPK) were shown to phosphorylate and decrease the activity of HNF4α. Activation of the ERK1/2 signalling pathway, inducing proliferation and survival, inhibits the expression of HNF4α. However, based on our previous results we hypothesized that HNF4α is also regulated at the post-transcriptional level by ERK1/2. Here we show that ERK1/2 is capable of directly phosphorylating HNF4α in vitro at several phosphorylation sites including residues previously shown to be targeted by other kinases, as well. Furthermore, we also demonstrate that phosphorylation of HNF4α leads to a reduced trans-activational capacity of the nuclear receptor in luciferase reporter gene assay. We confirm the functional relevance of these findings by demonstrating with ChIP-qPCR experiments that 30-minute activation of ERK1/2 leads to reduced chromatin binding of HNF4α. Accordingly, we have observed decreasing but not disappearing binding of HNF4α to the target genes. In addition, 24-hour activation of the pathway further decreased HNF4α chromatin binding to specific loci in ChIP-qPCR experiments, which confirms the previous reports on the decreased expression of the HNF4a gene due to ERK1/2 activation. Our data suggest that the ERK1/2 pathway plays an important role in the regulation of HNF4α-dependent hepatic gene expression. PMID:28196117

  16. Overview of the International R&D Recycling Activities of the Nuclear Fuel Cycle

    SciTech Connect

    Patricia Paviet-Hartmann

    2012-10-01

    Nuclear power has demonstrated over the last 30 years its capacity to produce base-load electricity at a low, predictable and stable cost due to the very low economic dependence on the price of uranium. However the management of used nuclear fuel remains the “Achilles’ Heel” of this energy source since the storage of used nuclear fuel is increasing as evidenced by the following number with 2,000 tons of UNF produced each year by the 104 US nuclear reactor units which equates to a total of 62,000 spent fuel assemblies stored in dry cask and 88,000 stored in pools. Two options adopted by several countries will be presented. The first one adopted by Europe, Japan and Russia consists of recycling the used nuclear fuel after irradiation in a nuclear reactor. Ninety six percent of uranium and plutonium contained in the spent fuel could be reused to produce electricity and are worth recycling. The separation of uranium and plutonium from the wastes is realized through the industrial PUREX process so that they can be recycled for re-use in a nuclear reactor as a mixed oxide (MOX) fuel. The second option undertaken by Finland, Sweden and the United States implies the direct disposal of used nuclear fuel into a geologic formation. One has to remind that only 30% of the worldwide used nuclear fuel are currently recycled, the larger part being stored (90% in pool) waiting for scientific or political decisions. A third option is emerging with a closed fuel cycle which will improve the global sustainability of nuclear energy. This option will not only decrease the volume amount of nuclear waste but also the long-term radiotoxicity of the final waste, as well as improving the long-term safety and the heat-loading of the final repository. At the present time, numerous countries are focusing on the R&D recycling activities of the ultimate waste composed of fission products and minor actinides (americium and curium). Several new chemical extraction processes, such as TRUSPEAK

  17. Multiple phosphorylation sites at the C-terminus regulate nuclear import of HCMV DNA polymerase processivity factor ppUL44

    SciTech Connect

    Alvisi, Gualtiero; Marin, Oriano; Pari, Gregory; Mancini, Manuela; Avanzi, Simone; Loregian, Arianna; Jans, David A.; Ripalti, Alessandro

    2011-09-01

    The processivity factor of human cytomegalovirus DNA polymerase, phosphoprotein ppUL44, is essential for viral replication. During viral infection ppUL44 is phosphorylated by the viral kinase pUL97, but neither the target residues on ppUL44 nor the effect of phosphorylation on ppUL44's activity are known. We report here that ppUL44 is phosphorylated when transiently expressed in mammalian cells and coimmunoprecipitates with cellular kinases. Of three potential phosphorylation sites (S413, S415, S418) located upstream of ppUL44's nuclear localization signal (NLS) and one (T427) within the NLS itself, protein kinase CK2 (CK2) specifically phosphorylates S413, to trigger a cascade of phosphorylation of S418 and S415 by CK1 and CK2, respectively. Negative charge at the CK2/CK1 target serine residues facilitates optimal nuclear accumulation of ppUL44, whereas negative charge on T427, a potential cyclin-dependent 1 phosphorylation site, strongly decreases nuclear accumulation. Thus, nuclear transport of ppUL44 is finely tuned during viral infection through complex phosphorylation events.

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

    PubMed Central

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

    2016-01-01

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

  19. Improving importance estimation in pool-based batch active learning for approximate linear regression.

    PubMed

    Kurihara, Nozomi; Sugiyama, Masashi

    2012-12-01

    Pool-based batch active learning is aimed at choosing training inputs from a 'pool' of test inputs so that the generalization error is minimized. P-ALICE (Pool-based Active Learning using Importance-weighted least-squares learning based on Conditional Expectation of the generalization error) is a state-of-the-art method that can cope with model misspecification by weighting training samples according to the importance (i.e., the ratio of test and training input densities). However, importance estimation in the original P-ALICE is based on the assumption that the number of training samples to gather is small, which is not always true in practice. In this paper, we propose an alternative scheme for importance estimation based on the inclusion probability, and show its validity through numerical experiments.

  20. Ribonucleotide reductase activity is coupled to DNA synthesis via proliferating cell nuclear antigen.

    PubMed

    Salguero, Israel; Guarino, Estrella; Shepherd, Marianne E A; Deegan, Tom D; Havens, Courtney G; MacNeill, Stuart A; Walter, Johannes C; Kearsey, Stephen E

    2012-04-24

    Synthesis of deoxynucleoside triphosphates (dNTPs) is required for both DNA replication and DNA repair and is catalyzed by ribonucleotide reductases (RNR), which convert ribonucleotides to their deoxy forms [1, 2]. Maintaining the correct levels of dNTPs for DNA synthesis is important for minimizing the mutation rate [3-7], and this is achieved by tight regulation of RNR [2, 8, 9]. In fission yeast, RNR is regulated in part by a small protein inhibitor, Spd1, which is degraded in S phase and after DNA damage to allow upregulation of dNTP supply [10-12]. Spd1 degradation is mediated by the activity of the CRL4(Cdt2) ubiquitin ligase complex [5, 13, 14]. This has been reported to be dependent on modulation of Cdt2 levels, which are cell cycle regulated, peaking in S phase, and which also increase after DNA damage in a checkpoint-dependent manner [7, 13]. We show here that Cdt2 level fluctuations are not sufficient to regulate Spd1 proteolysis and that the key step in this event is the interaction of Spd1 with the polymerase processivity factor proliferating cell nuclear antigen (PCNA), complexed onto DNA. This mechanism thus provides a direct link between DNA synthesis and RNR regulation.

  1. Transcriptional integration of metabolism by the nuclear sterol-activated receptors LXR and FXR.

    PubMed

    Calkin, Anna C; Tontonoz, Peter

    2012-03-14

    Nuclear receptors are integrators of hormonal and nutritional signals, mediating changes to metabolic pathways within the body. Given that modulation of lipid and glucose metabolism has been linked to diseases including type 2 diabetes, obesity and atherosclerosis, a greater understanding of pathways that regulate metabolism in physiology and disease is crucial. The liver X receptors (LXRs) and the farnesoid X receptors (FXRs) are activated by oxysterols and bile acids, respectively. Mounting evidence indicates that these nuclear receptors have essential roles, not only in the regulation of cholesterol and bile acid metabolism but also in the integration of sterol, fatty acid and glucose metabolism.

  2. Transcriptional integration of metabolism by the nuclear sterol-activated receptors LXR and FXR

    PubMed Central

    2013-01-01

    Nuclear receptors are integrators of hormonal and nutritional signals, mediating changes to metabolic pathways within the body. Given that modulation of lipid and glucose metabolism has been linked to diseases including type 2 diabetes, obesity and atherosclerosis, a greater understanding of pathways that regulate metabolism in physiology and disease is crucial. The liver X receptors (LXRs) and the farnesoid X receptors (FXRs) are activated by oxysterols and bile acids, respectively. Mounting evidence indicates that these nuclear receptors have essential roles, not only in the regulation of cholesterol and bile acid metabolism but also in the integration of sterol, fatty acid and glucose metabolism. PMID:22414897

  3. Antifungal activity of different neem leaf extracts and the nimonol against some important human pathogens

    PubMed Central

    Mahmoud, D.A.; Hassanein, N.M.; Youssef, K.A.; Abou Zeid, M.A.

    2011-01-01

    This study was conducted to evaluate the effect of aqueous, ethanolic and ethyl acetate extracts from neem leaves on growth of some human pathogens (Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Aspergillus terreus, Candida albicans and Microsporum gypseum) in vitro. Different concentrations (5, 10, 15 and 20%) prepared from these extracts inhibited the growth of the test pathogens and the effect gradually increased with concentration. The 20% ethyl acetate extract gave the strongest inhibition compared with the activity obtained by the same concentration of the other extracts. High Performance Liquid Chromatography (HPLC) analysis of ethyl acetate extract showed the presence of a main component (nimonol) which was purified and chemically confirmed by Nuclear Magnetic Resonance (NMR) spectroscopic analysis. The 20% ethyl acetate extract lost a part of its antifungal effect after pooling out the nimonol and this loss in activity was variable on test pathogens. The purified nimonol as a separate compound did not show any antifungal activity when assayed against all the six fungal pathogens. PMID:24031718

  4. Changes in the nuclear protein kinase activities in the regenerating liver of partially irradiated rat

    SciTech Connect

    Asami, K.; Kobayashi, H.; Fujiwara, A.; Yasumasu, I. )

    1989-09-01

    X rays (4.8 Gy) inhibit both DNA synthesis and phosphorylation of histone H1 in the regenerating liver of the rat. To determine the cause of the inhibition of histone H1 phosphorylation, changes in the nuclear protein kinase activities during the prereplicative phase of regeneration were measured. The cAMP-dependent protein kinase activity was low during regeneration, and the changes in the activity were not statistically significant. The cAMP-independent protein kinase activity increased at 15 h, decreased at 18 h, and increased again at 24 h after partial hepatectomy. X irradiation prior to partial hepatectomy did not inhibit the increase at 15 h, but it did inhibit the increase at 24 h. The activity was not inhibited by isoquinolinesulfonamide inhibitors such as H-7, and it was activated by a commercial preparation of an inhibitor protein of the cAMP-dependent kinase. It was also inhibited by quercetin. The possibility that the radiation-sensitive nuclear protein kinase is a nuclear cAMP-independent protein kinase specific for histone H1 is considered.

  5. III. The importance of physical activity and aerobic fitness for cognitive control and memory in children.

    PubMed

    Chaddock-Heyman, Laura; Hillman, Charles H; Cohen, Neal J; Kramer, Arthur F

    2014-12-01

    In this chapter, we review literature that examines the association among physical activity, aerobic fitness, cognition, and the brain in elementary school children (ages 7-10 years). Specifically, physical activity and higher levels of aerobic fitness in children have been found to benefit brain structure, brain function, cognition, and school achievement. For example, higher fit children have larger brain volumes in the basal ganglia and hippocampus, which relate to superior performance on tasks of cognitive control and memory, respectively, when compared to their lower fit peers. Higher fit children also show superior brain function during tasks of cognitive control, better scores on tests of academic achievement, and higher performance on a real-world street crossing task, compared to lower fit and less active children. The cross-sectional findings are strengthened by a few randomized, controlled trials, which demonstrate that children randomly assigned to a physical activity intervention group show greater brain and cognitive benefits compared to a control group. Because these findings suggest that the developing brain is plastic and sensitive to lifestyle factors, we also discuss typical structural and functional brain maturation in children to provide context in which to interpret the effects of physical activity and aerobic fitness on the developing brain. This research is important because children are becoming increasingly sedentary, physically inactive, and unfit. An important goal of this review is to emphasize the importance of physical activity and aerobic fitness for the cognitive and brain health of today's youth.

  6. Activating the nuclear piston mechanism of 3D migration in tumor cells.

    PubMed

    Petrie, Ryan J; Harlin, Heather M; Korsak, Lulu I T; Yamada, Kenneth M

    2017-01-02

    Primary human fibroblasts have the remarkable ability to use their nucleus like a piston, switching from low- to high-pressure protrusions in response to the surrounding three-dimensional (3D) matrix. Although migrating tumor cells can also change how they migrate in response to the 3D matrix, it is not clear if they can switch between high- and low-pressure protrusions like primary fibroblasts. We report that unlike primary fibroblasts, the nuclear piston is not active in fibrosarcoma cells. Protease inhibition rescued the nuclear piston mechanism in polarized HT1080 and SW684 cells and generated compartmentalized pressure. Achieving compartmentalized pressure required the nucleoskeleton-cytoskeleton linker protein nesprin 3, actomyosin contractility, and integrin-mediated adhesion, consistent with lobopodia-based fibroblast migration. In addition, this activation of the nuclear piston mechanism slowed the 3D movement of HT1080 cells. Together, these data indicate that inhibiting protease activity during polarized tumor cell 3D migration is sufficient to restore the nuclear piston migration mechanism with compartmentalized pressure characteristic of nonmalignant cells.

  7. Experimental Observations of Nuclear Activity in Deuterated Materials Subjected to a Low-Energy Photon Beam

    NASA Technical Reports Server (NTRS)

    Steinetz, Bruce M.; Benyo, Theresa L.; Pines, Vladimir; Pines, Marianna; Forsley, Lawrence P.; Westmeyer, Paul A.; Chait, Arnon; Becks, Michael D.; Martin, Richard E.; Hendricks, Robert C.; Penney, Nicholas; Marsolais, Annette M.; Kamm, Tracy R.

    2017-01-01

    Exposure of highly deuterated materials to a low-energy (nom. 2 MeV) photon beam resulted in nuclear activity of both the parent metals of hafnium and erbium and a witness material (molybdenum) mixed with the reactants. Gamma spectral analysis of all deuterated materials, ErD2.8+C36D74+Mo and HfD2+C36D74+Mo, showed that nuclear processes had occurred as shown by unique gamma signatures. For the deuterated erbium specimens, posttest gamma spectra showed evidence of radioisotopes of erbium ((163)Er and (171)Er) and of molybdenum ((99)Mo and (101)Mo) and by beta decay, technetium (99mTc and 101Tc). For the deuterated hafnium specimens, posttest gamma spectra showed evidence of radioisotopes of hafnium (180mHf and 181Hf) and molybdenum ((99)Mo and (101)Mo), and by beta decay, technetium ((99m)Tc and (101)Tc). In contrast, when either the hydrogenated or non-gas-loaded erbium or hafnium materials were exposed to the gamma flux, the gamma spectra revealed no new isotopes. Neutron activation materials showed evidence of thermal and epithermal neutrons. CR-39 solid-state nuclear track detectors showed evidence of fast neutrons with energies between 1.4 and 2.5 MeV and several instances of triple tracks, indicating (is) greater than 10 MeV neutrons. Further study is required to determine the mechanism causing the nuclear activity.

  8. Designing ARIES-CS compact radial build and nuclear system: Neutronics, shielding, and activation

    SciTech Connect

    El-GuebalyUniv. Wisco, L.; Wilson, P.; Sawan, M.; Sviatoslavsky, G.; Slaybaugh, R; Kiedrowski, B; Ibrahim, A.; MartinUniv Wiscons, C.; Tautges, Tim; Raffray, R.; Lyon, J.; Wang, X.; Bromberg, L.; Merrill, Brad; Wagner, L.; Najmabadi, F.

    2008-01-01

    Within the ARIES-CS project, design activities have focused on developing the first compact device that enhances the attractiveness of the stellarator as a power plant. The objectives of this paper are to review the nuclear elements that received considerable attention during the design process and provide a perspective on their successful integration into the final design. Among these elements are the radial build definition, the well-optimized in-vessel components that satisfy the ARIES top-level requirements, the carefully selected nuclear and engineering parameters to produce an economic optimum, the modeling for the first time ever-of the highly complex stellarator geometry for the three-dimensional nuclear assessment, and the overarching safety and environmental constraints to deliver an attractive, reliable, and truly compact stellarator power plant.

  9. Safety of evolutionary and innovative nuclear reactors: IAEA activities and world efforts

    SciTech Connect

    Saito, T.; Gasparini, M.

    2004-07-01

    'Defence in Depth' approach constitutes the basis of the IAEA safety standards for nuclear power plants. Lessons learned from the current generation of reactors suggest that, for the next generation of reactor designs, the Defence in Depth philosophy should be retained, and that its implementation should be guided by the probabilistic insights. Recent developments in the area of general safety requirements based on Defence in Depth approach are examined and summarized. Global efforts to harmonize safety requirements for evolutionary nuclear power plants have involved many countries and organizations such as IAEA, US EPRI and European Utility EUR Organization. In recent years, developments of innovative nuclear power plants are also being discussed. The IAEA is currently developing a safety approach specifically for innovative nuclear reactors. This approach will eventually lead to a proposal of safety requirements for innovative reactors. Such activities related to safety requirements of evolutionary and innovative reactors are introduced. Various evolutionary and innovative reactor designs are reported in the world. The safety design features of evolutionary large LWRs, innovative LWRs, Modular High Temperature Gas Reactors and Small Liquid Metal Cooled LMRs are also introduced. Enhanced safety features proposed in such reactors are discussed and summarized according to the levels of Defence in Depth. For future nuclear plants, international cooperation and harmonization, especially in the area of safety, appear to be inevitable. Based on the past experience with many member states, the IAEA believes itself to be the uniquely positioned international organization to play this key role. (authors)

  10. How My Daughter Taught Me to Teach: The Importance of Active Communication

    ERIC Educational Resources Information Center

    Hunt-Gierut, Deborah

    2011-01-01

    In this article, the author shares how her daughter, who was diagnosed with a profound hearing loss when she was a year old, taught her to teach, and demonstrates the importance of active communication. Teaching her daughter English as her second language has posed many challenges, but has also revealed successful strategies that the author has…

  11. 77 FR 19681 - Agency Information Collection Activities: Importers of Merchandise Subject to Actual Use Provisions

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-02

    ... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Importers of Merchandise Subject to Actual Use Provisions AGENCY: U.S. Customs and Border Protection, Department of... collection. SUMMARY: U.S. Customs and Border Protection (CBP) of the Department of Homeland Security will...

  12. 77 FR 55486 - Agency Information Collection Activities: Importer ID Input Record

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-10

    ... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Importer ID Input Record AGENCY: U.S. Customs and Border Protection, Department of Homeland Security. ACTION: 30-Day Notice and request for comments; Extension of an existing collection of information. SUMMARY: U.S....

  13. Standardization of Administered Activities in Pediatric Nuclear Medicine: A Report of the First Nuclear Medicine Global Initiative Project, Part 2-Current Standards and the Path Toward Global Standardization.

    PubMed

    Fahey, Frederic H; Bom, Henry Hee-Seung; Chiti, Arturo; Choi, Yun Young; Huang, Gang; Lassmann, Michael; Laurin, Norman; Mut, Fernando; Nuñez-Miller, Rodolfo; O'Keeffe, Darin; Pradhan, Prasanta; Scott, Andrew M; Song, Shaoli; Soni, Nischal; Uchiyama, Mayuki; Vargas, Luis

    2016-07-01

    The Nuclear Medicine Global Initiative (NMGI) was formed in 2012 and consists of 13 international organizations with direct involvement in nuclear medicine. The underlying objectives of the NMGI are to promote human health by advancing the field of nuclear medicine and molecular imaging, encourage global collaboration in education, and harmonize procedure guidelines and other policies that ultimately lead to improvements in quality and safety in the field throughout the world. For its first project, the NMGI decided to consider the issues involved in the standardization of administered activities in pediatric nuclear medicine. It was decided to divide the final report of this project into 2 parts. Part 1 was published in this journal in the spring of 2015. This article presents part 2 of the final report. It discusses current standards for administered activities in children and adolescents that have been developed by various professional organizations. It also presents an evaluation of the current practice of pediatric nuclear medicine specifically with regard to administered activities as determined by an international survey of 313 nuclear medicine clinics and centers from 29 countries. Lastly, it provides recommendations for a path toward global standardization of the administration of radiopharmaceuticals in children.

  14. The importance of work or productive activity in life care planning and case management

    PubMed Central

    Reid, Christine; Riddick-Grisham, Susan

    2015-01-01

    Abstract INTRODUCTION: The importance of work or productive activity for the well-being, community integration, and quality of life of people living with disabilities is addressed, with implications for life care planning and case management. BACKGROUND: The role of work or productive activity in our society, and consequences of deprivation if rehabilitation services do not address vocational effects of disabilities, is explored. A continuum of productivity options is introduced; types of vocational rehabilitation assessment processes and interventions are described. PURPOSE: The role of vocational rehabilitation services in life care planning and case management is discussed, focusing on quality of life for people living with disabilities. CONCLUSION: Rehabilitation and health care professionals should understand the importance of work or other productive activity, and support the development of appropriate plans to address those needs among people who have disabilities. PMID:26409330

  15. Investigating the Nuclear Activity of Barred Spiral Galaxies: The Case of NGC 1672

    NASA Technical Reports Server (NTRS)

    Jenkins, L. P.; Brandt, W. N.; Colbert, E. J.; Koribalski, B.; Kuntz, K. D.; Levan, A. J.; Ojha, R.; Roberts, T. P.; Ward, M. J.; Zezas, A.

    2011-01-01

    We have performed an X-ray study of the nearby barred spiral galaxy NGC 1672, primarily to ascertain the effect of the bar on its nuclear activity. We use both Chandra and XMM-Newton observations to investigate its X-ray properties, together with supporting high-resolution optical imaging data from the Hubble Space Telescope (HST) infrared imaging from the Spitzer Space Telescope, and Australia Telescope Compact Array ground-based radio data. We detect 28 X-ray sources within the D25 area of the galaxy; many are spatially correlated with star formation in the bar and spiral arms, and two are identified as background galaxies in the HST images. Nine of the X-ray sources are ultraluminous X-ray sources, with the three brightest (LX 5 * 10(exp 39) erg s(exp -1)) located at the ends of the bar. With the spatial resolution of Chandra, we are able to show for the first time that NGC 1672 possesses a hard (1.5) nuclear X-ray source with a 2-10 keV luminosity of 4 * 10(exp 38) erg s(exp -1). This is surrounded by an X-ray-bright circumnuclear star-forming ring, comprised of point sources and hot gas, which dominates the 2-10 keV emission in the central region of the galaxy. The spatially resolved multiwavelength photometry indicates that the nuclear source is a low-luminosity active galactic nucleus (LLAGN), but with star formation activity close to the central black hole. A high-resolution multiwavelength survey is required to fully assess the impact of both large-scale bars and smaller-scale phenomena such as nuclear bars, rings, and nuclear spirals on the fueling of LLAGN.

  16. Nuclear F-actin enhances the transcriptional activity of β-catenin by increasing its nuclear localization and binding to chromatin.

    PubMed

    Yamazaki, Shota; Yamamoto, Koji; de Lanerolle, Primal; Harata, Masahiko

    2016-04-01

    Actin plays multiple roles both in the cytoplasm and in the nucleus. Cytoplasmic actin, in addition to its structural role in the cytoskeleton, also contributes to the subcellular localization of transcription factors by interacting with them or their partners. The transcriptional cofactor β-catenin, which acts as an intracellular transducer of canonical Wnt signaling, indirectly associates with the cytoplasmic filamentous actin (F-actin). Recently, it has been observed that F-actin is transiently formed within the nucleus in response to serum stimulation and integrin signaling, and also during gene reprogramming. Despite these earlier observations, information about the function of nuclear F-actin is poorly defined. Here, by facilitating the accumulation of nuclear actin artificially, we demonstrate that polymerizing nuclear actin enhanced the nuclear accumulation and transcriptional function of β-catenin. Our results also show that the nuclear F-actin colocalizes with β-catenin and enhances the binding of β-catenin to the downstream target genes of the Wnt/β-catenin signaling pathway, including the genes for the cell cycle regulators c-myc and cyclin D, and the OCT4 gene. Nuclear F-actin itself also associated with these genes. Since Wnt/β-catenin signaling has important roles in cell differentiation and pluripotency, our observations suggest that nuclear F-actin formed during these biological processes is involved in regulating Wnt/β-catenin signaling.

  17. Rational engineering of L-asparaginase reveals importance of dual activity for cancer cell toxicity.

    PubMed

    Offman, Marc N; Krol, Marcin; Patel, Naina; Krishnan, Shekhar; Liu, JiZhong; Saha, Vaskar; Bates, Paul A

    2011-02-03

    Using proteins in a therapeutic context often requires engineering to modify functionality and enhance efficacy. We have previously reported that the therapeutic antileukemic protein macromolecule Escherichia coli L-asparaginase is degraded by leukemic lysosomal cysteine proteases. In the present study, we successfully engineered L-asparaginase to resist proteolytic cleavage and at the same time improve activity. We employed a novel combination of mutant sampling using a genetic algorithm in tandem with flexibility studies using molecular dynamics to investigate the impact of lid-loop and mutations on drug activity. Applying these methods, we successfully predicted the more active L-asparaginase mutants N24T and N24A. For the latter, a unique hydrogen bond network contributes to higher activity. Furthermore, interface mutations controlling secondary glutaminase activity demonstrated the importance of this enzymatic activity for drug cytotoxicity. All selected mutants were expressed, purified, and tested for activity and for their ability to form the active tetrameric form. By introducing the N24A and N24A R195S mutations to the drug L-asparaginase, we are a step closer to individualized drug design.

  18. An Important Natural Genetic Resource of Oreochromis niloticus (Linnaeus, 1758) Threatened by Aquaculture Activities in Loboi Drainage, Kenya

    PubMed Central

    Ndiwa, Titus Chemandwa; Nyingi, Dorothy Wanja; Agnese, Jean-François

    2014-01-01

    The need to improve food security in Africa through culture of tilapias has led to transfer of different species from their natural ranges causing negative impacts on wild fish genetic resources. Loboi swamp in Kenya is fed by three hot springs: Lake Bogoria Hotel, Chelaba and Turtle Springs, hosting natural populations of Oreochromis niloticus. The present study aimed at better genetic characterization of these threatened populations. Partial mtDNA sequences of the D-loop region and variations at 16 microsatellite loci were assessed in the three hot spring populations and compared with three other natural populations of O. niloticus in the region. Results obtained indicated that the hot spring populations had mitochondrial and nuclear genetic variability similar to or higher than the large closely related populations. This may be attributed to the perennial nature of the hot springs, which do not depend on rainfall but rather receive permanent water supply from deep aquifers. The study also revealed that gene flow between the three different hot spring populations was sufficiently low thus allowing their differentiation. This differentiation was unexpected considering the very close proximity of the springs to each other. It is possible that the swamp creates a barrier to free movement of fish from one spring to the other thereby diminishing gene flow. Finally, the most surprising and worrying results were that the three hot spring populations are introgressed by mtDNA genes of O. leucostictus, while microsatellite analysis suggested that some nuclear genes may also have crossed the species barrier. It is very likely that the recent intensification of aquaculture activities in the Loboi drainage may be responsible for these introgressions. Taking into account the importance of these new genetic resources, protection and management actions of the Loboi swamp should be accorded top priority to prevent the loss of these spring populations. PMID:25222491

  19. An important natural genetic resource of Oreochromis niloticus (Linnaeus, 1758) threatened by aquaculture activities in Loboi drainage, Kenya.

    PubMed

    Ndiwa, Titus Chemandwa; Nyingi, Dorothy Wanja; Agnese, Jean-François

    2014-01-01

    The need to improve food security in Africa through culture of tilapias has led to transfer of different species from their natural ranges causing negative impacts on wild fish genetic resources. Loboi swamp in Kenya is fed by three hot springs: Lake Bogoria Hotel, Chelaba and Turtle Springs, hosting natural populations of Oreochromis niloticus. The present study aimed at better genetic characterization of these threatened populations. Partial mtDNA sequences of the D-loop region and variations at 16 microsatellite loci were assessed in the three hot spring populations and compared with three other natural populations of O. niloticus in the region. Results obtained indicated that the hot spring populations had mitochondrial and nuclear genetic variability similar to or higher than the large closely related populations. This may be attributed to the perennial nature of the hot springs, which do not depend on rainfall but rather receive permanent water supply from deep aquifers. The study also revealed that gene flow between the three different hot spring populations was sufficiently low thus allowing their differentiation. This differentiation was unexpected considering the very close proximity of the springs to each other. It is possible that the swamp creates a barrier to free movement of fish from one spring to the other thereby diminishing gene flow. Finally, the most surprising and worrying results were that the three hot spring populations are introgressed by mtDNA genes of O. leucostictus, while microsatellite analysis suggested that some nuclear genes may also have crossed the species barrier. It is very likely that the recent intensification of aquaculture activities in the Loboi drainage may be responsible for these introgressions. Taking into account the importance of these new genetic resources, protection and management actions of the Loboi swamp should be accorded top priority to prevent the loss of these spring populations.

  20. 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.

  1. 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

  2. c-Cbl mediates the degradation of tumorigenic nuclear β-catenin contributing to the heterogeneity in Wnt activity in colorectal tumors

    PubMed Central

    Shashar, Moshe; Siwak, Jamaica; Tapan, Umit; Lee, Shin Yin; Meyer, Rosana D.; Parrack, Paige; Tan, Josenia; Khatami, Fatemeh; Francis, Jean; Zhao, Qing; Hartshorn, Kevan; Kolachalama, Vijaya B.; Rahimi, Nader; Chitalia, Vipul

    2016-01-01

    Despite the loss of Adenomatous Polyposis Coli (APC) in a majority of colorectal cancers (CRC), not all CRCs bear hallmarks of Wnt activation, such as nuclear β-catenin. This underscores the presence of other Wnt regulators that are important to define, given the pathogenic and prognostic roles of nuclear β-catenin in human CRC. Herein, we investigated the effect of Casitas B-lineage lymphoma (c-Cbl) on nuclear β-catenin, which is an oncoprotein upregulated in CRC due to loss-of-function APC or gain-of-function CTNNB1 mutations. Despite mechanistic rationale and recent discoveries of c-Cbl's mutations in solid tumors, little is known about its functional importance in CRC. Our study in a cohort of human CRC patients demonstrated an inverse correlation between nuclear β-catenin and c-Cbl. Further investigation showed that the loss of c-Cbl activity significantly enhanced nuclear β-catenin and CRC tumor growth in cell culture and a mouse xenograft model. c-Cbl interacted with and downregulated β-catenin in a manner that was independent of CTNNB1 or APC mutation status. This study demonstrates a previously unrecognized function of c-Cbl as a negative regulator of CRC. PMID:27661103

  3. Towards novel efficient and stable nuclear import signals: synthesis and properties of trimethylguanosine cap analogs modified within the 5',5'-triphosphate bridge.

    PubMed

    Zytek, Malgorzata; Kowalska, Joanna; Lukaszewicz, Maciej; Wojtczak, Blazej A; Zuberek, Joanna; Ferenc-Mrozek, Aleksandra; Darzynkiewicz, Edward; Niedzwiecka, Anna; Jemielity, Jacek

    2014-12-07

    A trimethylguanosine (TMG) cap is present at the 5' end of several small nuclear and nucleolar RNAs. Recently, it has been reported that the TMG cap is a potential nuclear import signal for nucleus-targeting therapeutic nucleic acids and proteins. The import is mediated by recognition of the TMG cap by the snRNA transporting protein, snurportin1. This work describes the synthesis and properties of a series of dinucleotide TMG cap (m3(2,2,7)GpppG) analogs modified in the 5',5'-triphosphate bridge as tools to study TMG cap-dependent biological processes. The bridge was altered at different positions by introducing either bridging (imidodiphosphate, O to NH and methylenebisphosphonate, O to CH2) or non-bridging (phosphorothioate, O to S and boranophosphate, O to BH3) modifications, or by elongation to tetraphosphate. The stability of novel analogs in blood serum was studied to reveal that the α,β-bridging O to NH substitution (m3(2,2,7)GppNHpG) confers the highest resistance. Short RNAs capped with analogs containing α,β-bridging (m3(2,2,7)GppNHpG) or β-non-bridging (m3(2,2,7)GppSpG D2) modifications were resistant to decapping pyrophosphatase, hNudt16. Preliminary studies on binding by human snurportin1 revealed that both O to NH and O to S substitutions support this binding. Due to favorable properties in all three assays, m3(2,2,7)GppNHpG was selected as a promising candidate for further studies on the efficiency of the TMG cap as a nuclear import signal.

  4. The importance of domain closure for the auto-activation of ERK2

    PubMed Central

    Barr, Daniel; Oashi, Taiji; Burkhard, Kimberly; Lucius, Sarah; Samadani, Ramin; Zhang, Jun; Shapiro, Paul; MacKerell, Alexander D.; van der Vaart, Arjan

    2011-01-01

    Extracellular signal-regulated kinases-1 and 2 (ERK1/2) play a critical role in regulating cell division and have been implicated in cancer. In addition to activation by the MAPK/ERK kinases 1 and 2 (MEK1/2), certain mutants of ERK2 can be activated by auto-phosphorylation. To identify the mechanism of auto-activation, we have performed a series of molecular dynamics simulations of ERK1/2 in various stages of activation as well as the constitutively active Q103A, I84A, L73P and R65S ERK2 mutants. Our simulations indicate the importance of domain closure for auto-activation and activity regulation, with that event occurring prior to folding of the activation lip and of loop L16. Results indicate that the second phosphorylation event to T183 disrupts hydrogen bonding involving D334 thereby allowing the kinase to lock into the active conformation. Based on the simulations, three predictions were made: G83A was suggested to impede activation, K162M was suggested to perturb the interface between the N and C-domain leading to activation, and Q64C was hypothesized to stop folding of loop L16 thereby perturbing the homodimerization interface. Functional analysis of the mutants validated the predictions concerning the G83A and Q64C mutants. The K162M mutant did not autoactivate as predicted however, which may be due to the location of the residue on the protein surface near the ED substrate docking domain. PMID:21842857

  5. Mast cells enhance T cell activation: Importance of mast cell-derived TNF

    NASA Astrophysics Data System (ADS)

    Nakae, Susumu; Suto, Hajime; Kakurai, Maki; Sedgwick, Jonathon D.; Tsai, Mindy; Galli, Stephen J.

    2005-05-01

    Mast cells are not only important effector cells in immediate hypersensitivity reactions and immune responses to pathogens but also can contribute to T cell-mediated disorders. However, the mechanisms by which mast cells might influence T cells in such settings are not fully understood. We find that mast cells can enhance proliferation and cytokine production in multiple T cell subsets. Mast cell-dependent enhancement of T cell activation can be promoted by FcRI-dependent mast cell activation, TNF production by both mast cells and T cells, and mast cell-T cell contact. However, at high concentrations of cells, mast cells can promote T cell activation independent of IgE or TNF. Finally, mast cells also can promote T cell activation by means of soluble factors. These findings identify multiple mechanisms by which mast cells can influence T cell proliferation and cytokine production. allergy | asthma | autoimmunity | cytokines | immune response

  6. Porcine circovirus type 2 induces the activation of nuclear factor kappa B by I{kappa}B{alpha} degradation

    SciTech Connect

    Wei Li; Kwang, Jimmy; Wang Jin; Shi Lei; Yang Bing; Li Yongqing; Liu Jue

    2008-08-15

    The transcription factor NF-{kappa}B is commonly activated upon virus infection and a key player in the induction and regulation of the host immune response. The present study demonstrated for the first time that porcine circovirus type 2 (PCV2), which is the primary causative agent of an emerging swine disease, postweaning multisystemic wasting syndrome, can activate NF-{kappa}B in PCV2-infected PK15 cells. In PCV2-infected cells, NF-{kappa}B was activated concomitantly with viral replication, which was characterized by increased DNA binding activity, translocation of NF-{kappa}B p65 from the cytoplasm to the nucleus, as well as degradation and phosphorylation of I{kappa}B{alpha} protein. We further demonstrated PCV2-induced activation of NF-{kappa}B and colocalization of p65 nuclear translocation with virus replication in cultured cells. Treatment of cells with CAPE, a selective inhibitor of NF-{kappa}B activation, reduced virus protein expression and progeny production followed by decreasing PCV2-induced apoptotic caspase activity, indicating the involvement of this transcription factor in induction of cell death. Taken together, these data suggest that NF-{kappa}B activation is important for PCV2 replication and contributes to virus-mediated changes in host cells. The results presented here provide a basis for understanding molecular mechanism of PCV2 infection.

  7. The importance of myeloperoxidase enzyme activity in the pathogenesis of Crimean-Congo haemorrhagic fever.

    PubMed

    Guven, F M K; Aydin, H; Yildiz, G; Engin, A; Celik, V K; Bakir, D; Deveci, K

    2013-03-01

    Crimean-Congo haemorrhagic fever (CCHF) is a disease with a severe course including acute viral haemorrhagic fever, ecchymosis, thrombocytopenia, hepatic function disorder and high mortality. Myeloperoxidase (MPO) is an enzyme located in neutrophil granulocytes and plays an important role in the destruction of phagocytosed micro-organisms. The aim of this study was to analyse MPO enzyme activity in CCHF cases compared with a control group. A total of 47 randomly selected CCHF patients admitted to the Department of Infectious Diseases of Cumhuriyet University Hospital in Sivas, Turkey, were studied, and as a control group, 41 age- and sex-matched individuals without any systemic disease were included in this study. MPO enzyme activity was measured in plasma and leukocytes for both groups by the ELISA method. MPO plasma and MPO leukocyte values were calculated as 57.62 ± 8.85 and 44.84 ± 9.71 in CCHF patients, and 0.79 ± 0.29 and 0.49 ± 0.11 in the controls, respectively. MPO enzyme activity was statistically significantly higher in patients with CCHF when compared to the control group. In conclusion, MPO enzyme activity is directly related to the activation of phagocytic leukocytes, and increases in both the plasma and leukocytes in CCHF patients. The increase of the MPO enzyme activity in leukocytes due to viral load leads to the destruction of the leukocyte. It is thought that MPO enzyme activity in plasma was higher in CCHF patients due to the destruction of leukocytes. MPO enzyme activity may be important in terms of the prognosis in patients with CCHF; however, more extensive studies are required on this subject.

  8. Importin-7 mediates memory consolidation through regulation of nuclear translocation of training-activated MAPK in Drosophila

    PubMed Central

    Li, Qian; Zhang, Xuchen; Liang, Xitong; Zhang, Fang; Wang, Lianzhang; Zhong, Yi

    2016-01-01

    Translocation of signaling molecules, MAPK in particular, from the cytosol to nucleus represents a universal key element in initiating the gene program that determines memory consolidation. Translocation mechanisms and their behavioral impact, however, remain to be determined. Here, we report that a highly conserved nuclear transporter, Drosophila importin-7 (DIM-7), regulates import of training-activated MAPK for consolidation of long-term memory (LTM). We show that silencing DIM-7 functions results in impaired LTM, whereas overexpression of DIM-7 enhances LTM. This DIM-7–dependent regulation of LTM is confined to a consolidation time window and in mushroom body neurons. Image data show that bidirectional alteration in DIM-7 expression results in proportional changes in the intensity of training-activated MAPK accumulated within the nuclei of mushroom body neurons during LTM consolidation. Such DIM-7–regulated nuclear accumulation of activated MAPK is observed only in the training specified for LTM induction and determines the amplitude, but not the time course, of memory consolidation. PMID:26929354

  9. The hypersensitive response to tomato leaf curl New Delhi virus nuclear shuttle protein is inhibited by transcriptional activator protein.

    PubMed

    Hussain, Mazhar; Mansoor, Shahid; Iram, Shazia; Zafar, Yusuf; Briddon, Rob W

    2007-12-01

    The hypersensitive response (HR) is a common feature of plant disease resistance reactions and a type of programmed cell death (PCD). Many pathogens are able to modulate pathways involved in cell death. In contrast to animal viruses, inhibitors of PCD activity have not been identified for plant-infecting viruses. Previously, we have reported that the nuclear shuttle protein (NSP) of Tomato leaf curl New Delhi virus (ToLCNDV) induces an HR in Nicotiana tabacum and Lycopersicon esculentum plants when expressed under the control of the Cauliflower mosaic virus 35S promoter. However, HR is not evident in plants infected with ToLCNDV, suggesting that the virus encodes a factor (or factors) that counters this response. Analysis of all ToLCNDV-encoded genes pinpointed the transcriptional activator protein (TrAP) as the factor mediating the anti-HR effect. Deletion mutagenesis showed the central region of TrAP, containing a zinc finger domain and nuclear localization signal, to be important in inhibiting the HR. These results demonstrate that TrAP counters HR-induced cell death, the first such activity identified for a plant-infecting virus.

  10. Nuclear Chemistry.

    ERIC Educational Resources Information Center

    Chemical and Engineering News, 1979

    1979-01-01

    Provides a brief review of the latest developments in nuclear chemistry. Nuclear research today is directed toward increased activity in radiopharmaceuticals and formation of new isotopes by high-energy, heavy-ion collisions. (Author/BB)

  11. Dry, portable calorimeter for nondestructive measurement of the activity of nuclear fuel

    DOEpatents

    Beyer, Norman S.; Lewis, Robert N.; Perry, Ronald B.

    1976-01-01

    The activity of a quantity of heat-producing nuclear fuel is measured rapidly, accurately and nondestructively by a portable dry calorimeter comprising a preheater, an array of temperature-controlled structures comprising a thermally guarded temperature-controlled oven, and a calculation and control unit. The difference between the amounts of electric power required to maintain the oven temperature with and without nuclear fuel in the oven is measured to determine the power produced by radioactive disintegration and hence the activity of the fuel. A portion of the electronic control system is designed to terminate a continuing sequence of measurements when the standard deviation of the variations of the amount of electric power required to maintain oven temperature is within a predetermined value.

  12. The line-emitting gas in active galaxies - A probe of the nuclear engine

    NASA Technical Reports Server (NTRS)

    Veilleux, Sylvain

    1993-01-01

    This paper reviews some of the basic questions regarding the structure of the engine powering active galactic nuclei (AGN), the nature of the interaction between the AGN and the host galaxy, and the origin and evolution of AGN. The study of the dynamics and physical characteristics of the line-emitting gas in these objects has proven fruitful in addressing many of these issues. Recent advances in optical and infrared detector technology combined with the development of superior ground-based instruments have produced efficient new tools for the study of the line-emitting gas on nuclear and Galactic scales. Programs which take advantage of two of these new techniques, Fabry-Perot imaging spectroscopy and infrared spectroscopy, are described in this paper. The origin of nuclear activity in galaxies is also addressed in a third project which aims at determining the nature of luminous infrared galaxies.

  13. Esterase in Imported Fire Ants, Solenopsis invicta and S. richteri (Hymenoptera: Formicidae): Activity, Kinetics and Variation

    PubMed Central

    Chen, J.; Rashid, T.; Feng, G.

    2014-01-01

    Solenopsis invicta and Solenopsis richteri are two closely related invasive ants native to South America. Despite their similarity in biology and behavior, S. invicta is a more successful invasive species. Toxic tolerance has been found to be important to the success of some invasive species. Esterases play a crucial role in toxic tolerance of insects. Hence, we hypothesized that the more invasive S. invicta would have a higher esterase activity than S. richteri. Esterase activities were measured for workers and male and female alates of both ant species using α-naphthyl acetate and β-naphthyl acetate as substrates. Esterase activities in S. invicta were always significantly higher than those in S. richteri supporting our hypothesis. In S. invicta, male alates had the highest esterase activities followed by workers then female alates for both substrates. In S. richetri, for α-naphthyl acetate, male alates had the highest activity followed by female alates then workers, while for β-naphthyl acetate, female alates had the highest activity followed by male alates then workers. For workers, S. richteri showed significantly higher levels of variation about the mean esterase activity than S. invicta. However, S. invicta showed significantly higher levels of variation in both female and male alates. PMID:25408118

  14. Effects of primary metabolites of organophosphate flame retardants on transcriptional activity via human nuclear receptors.

    PubMed

    Kojima, Hiroyuki; Takeuchi, Shinji; Van den Eede, Nele; Covaci, Adrian

    2016-03-14

    Organophosphate flame retardants (OPFRs) have been used in a wide variety of applications and detected in several environmental matrices, including indoor air and dust. Continuous human exposure to these chemicals is of growing concern. In this study, the agonistic and/or antagonistic activities of 12 primary OPFR-metabolites against ten human nuclear receptors were examined using cell-based transcriptional assays, and compared to those of their parent compounds. As a result, 3-hydroxylphenyl diphenyl phosphate and 4-hydroxylphenyl diphenyl phosphate showed more potent estrogen receptor α (ERα) and ERβ agonistic activity than did their parent, triphenyl phosphate (TPHP). In addition, these hydroxylated TPHP-metabolites also showed ERβ antagonistic activity at higher concentrations and exhibited pregnane X receptor (PXR) agonistic activity as well as androgen receptor (AR) and glucocorticoid receptor (GR) antagonistic activities at similar levels to those of TPHP. Bis(2-butoxyethyl) 3'-hydroxy-2-butoxyethyl phosphate and 2-hydroxyethyl bis(2-butoxyethyl) phosphate act as PXR agonists at similar levels to their parent, tris(2-butoxyethyl) phosphate. On the other hand, seven diester OPFR-metabolites and 1-hydroxy-2-propyl bis(1-chloro-2-propyl) phosphate did not show any receptor activity. Taken together, these results suggest that hydroxylated TPHP-metabolites show increased estrogenicity compared to the parent compound, whereas the diester OPFR-metabolites may have limited nuclear receptor activity compared to their parent triester OPFRs.

  15. HSP72 inhibits Smad3 activation and nuclear translocation in renal epithelial-to-mesenchymal transition.

    PubMed

    Zhou, Yi; Mao, Haiping; Li, Shu; Cao, Shirong; Li, Zhijian; Zhuang, Shougang; Fan, Jinjin; Dong, Xiuqing; Borkan, Steven C; Wang, Yihan; Yu, Xueqing

    2010-04-01

    Although heat shock protein 72 (HSP72) ameliorates renal tubulointerstitial fibrosis by inhibiting epithelial-to-mesenchymal transition (EMT), the underlying mechanism is unknown. Because Smad proteins transduce TGF-beta signaling from the cytosol to the nucleus and HSP72 assists in protein folding and facilitates nuclear translocation, we investigated whether HSP72 inhibits TGF-beta-induced EMT by modulating Smad expression, activation, and nuclear translocation. To evaluate the roles of distinct HSP72 structural domains in these processes, we constructed vectors that expressed wild-type HSP72 or mutants lacking either the peptide-binding domain (HSP72-DeltaPBD), which is responsible for substrate binding and refolding, or the nuclear localization signal (HSP72-DeltaNLS). Overexpression of wild-type HSP72 or HSP72-DeltaNLS inhibited TGF-beta1-induced EMT, but HSP72-DeltaPBD did not, suggesting a critical role for the PBD in this inhibition. HSP72 overexpression inhibited TGF-beta1-induced phosphorylation and nuclear translocation of Smad3 and p-Smad3, but not Smad2; these inhibitory effects required the PBD but not the NLS. Coimmunoprecipitation assays suggested a physical interaction between Smad3 and the PBD. siRNA knockdown of endogenous HSP72 enhanced both TGF-beta1-induced Smad3 phosphorylation and EMT and confirmed the interaction of HSP72 with both Smad3 and p-Smad3. In vivo, induction of HSP72 by geranylgeranylacetone suppressed Smad3 phosphorylation in renal tubular cells after unilateral ureteral obstruction. In conclusion, HSP72 inhibits EMT in renal epithelial cells primarily by exerting domain-specific effects on Smad3 activation and nuclear translocation.

  16. Antiviral activity of KR-23502 targeting nuclear export of influenza B virus ribonucleoproteins.

    PubMed

    Jang, Yejin; Lee, Hye Won; Shin, Jin Soo; Go, Yun Young; Kim, Chonsaeng; Shin, Daeho; Malpani, Yashwardhan; Han, Soo Bong; Jung, Young-Sik; Kim, Meehyein

    2016-10-01

    The spiro compound 5,6-dimethyl-3H,3'H-spiro(benzofuran-2,1'-isobenzofuran)-3,3'-dione (KR-23502) has antiviral activity against influenza A and more potently B viruses. The aim of this study is to elucidate its mechanism of action. Subcellular localization and time-course expression of influenza B viral proteins, nucleoprotein (NP) and matrix protein 1 (M1), showed that KR-23502 reduced their amounts within 5 h post-infection. Early steps of virus life cycle, including virus entry, nuclear localization of NP and viral RNA-dependent RNA replication, were not affected by KR-23502. Instead it interrupted a later event corresponding to nuclear export of NP and M1 proteins. Delivery of viral ribonucleoprotein (vRNP)-M1 complex has been known to be mediated by the viral nuclear export protein (NEP) through interaction with cellular chromosomal maintenance 1 (CRM1) protein. In this study, we experimentally demonstrated that the compound targets the nuclear export of vRNP. Moreover, a single mutation (aspartate to glycine) at amino acid position 54 in M1 [M1(D54G)] was detected after 18 passages in the presence of KR-23502 with a 2-fold increase in 50% effective concentration indicating that this compound has a relatively high genetic barrier to resistance. Interestingly, it was observed that proteasome-mediated degradation of M1(D54G) was attenuated by KR-23502. In conclusion, we suggest that KR-23502 shows its anti-influenza activity by downregulating NEP/CRM1-mediated nuclear export of influenza vRNP and M1. KR-23502 provides a core chemical skeleton for further structure-based design of novel antivirals against influenza viruses.

  17. Nuclear star formation activity and black hole accretion in nearby Seyfert galaxies

    SciTech Connect

    Esquej, P.; Alonso-Herrero, A.; Hernán-Caballero, A.; González-Martín, O.; Ramos Almeida, C.; Rodríguez Espinosa, J. M.; Roche, P.; Mason, R. E.; Díaz-Santos, T.; Levenson, N. A.; Aretxaga, I.; Packham, C.

    2014-01-01

    Recent theoretical and observational works indicate the presence of a correlation between the star-formation rate (SFR) and active galactic nucleus (AGN) luminosity (and, therefore, the black hole accretion rate, M-dot {sub BH}) of Seyfert galaxies. This suggests a physical connection between the gas-forming stars on kpc scales and the gas on sub-pc scales that is feeding the black hole. We compiled the largest sample of Seyfert galaxies to date with high angular resolution (∼0.''4-0.''8) mid-infrared (8-13 μm) spectroscopy. The sample includes 29 Seyfert galaxies drawn from the AGN Revised Shapley-Ames catalog. At a median distance of 33 Mpc, our data allow us to probe nuclear regions on scales of ∼65 pc (median value). We found no general evidence of suppression of the 11.3 μm polycyclic aromatic hydrocarbon (PAH) emission in the vicinity of these AGN, and we used this feature as a proxy for the SFR. We detected the 11.3 μm PAH feature in the nuclear spectra of 45% of our sample. The derived nuclear SFRs are, on average, five times lower than those measured in circumnuclear regions of 600 pc in size (median value). However, the projected nuclear SFR densities (median value of 22 M {sub ☉} yr{sup –1} kpc{sup –2}) are a factor of 20 higher than those measured on circumnuclear scales. This indicates that the SF activity per unit area in the central ∼65 pc region of Seyfert galaxies is much higher than at larger distances from their nuclei. We studied the connection between the nuclear SFR and M-dot {sub BH} and showed that numerical simulations reproduce our observed relation fairly well.

  18. Studies of neutron and proton nuclear activation in low-Earth orbit

    NASA Technical Reports Server (NTRS)

    Laird, C. E.

    1982-01-01

    The expected induced radioactivity of experimental material in low Earth orbit was studied for characteristics of activating particles such as cosmic rays, high energy Earth albedo neutrons, trapped protons, and secondary protons and neutrons. The activation cross sections for the production of long lived radioisotopes and other existing nuclear data appropriate to the study of these reactions were compiled. Computer codes which are required to calculate the expected activation of orbited materials were developed. The decreased computer code used to predict the activation of trapped protons of materials placed in the expected orbits of LDEF and Spacelab II. Techniques for unfolding the fluxes of activating particles from the measured activation of orbited materials are examined.

  19. Pseudorabies Virus pUL46 Induces Activation of ERK1/2 and Regulates Herpesvirus-Induced Nuclear Envelope Breakdown

    PubMed Central

    Schulz, Katharina S.; Liu, XueQiao; Klupp, Barbara G.; Granzow, Harald; Cohen, Jeffrey I.

    2014-01-01

    ABSTRACT Herpesvirus capsid morphogenesis occurs in the nucleus, while final maturation takes place in the cytosol, requiring translocation of capsids through the nuclear envelope. The nuclear egress complex, consisting of homologs of herpes simplex virus pUL31 and pUL34, is required for efficient nuclear egress via primary envelopment and de-envelopment. Recently, we described an alternative mode of nuclear escape by fragmentation of the nuclear envelope induced by replication-competent pUL31 and pUL34 deletion mutants of the alphaherpesvirus pseudorabies virus (PrV), which had been selected by serial passaging in cell culture. Both passaged viruses carry congruent mutations in seven genes, including UL46, which encodes one of the major tegument proteins. Herpesvirus pUL46 homologs have recently been shown to activate the PI3K-Akt and ERK1/2 signaling pathways, which are involved in regulation of mitosis and apoptosis. Since in uninfected cells fragmentation of the nuclear envelope occurs during mitosis and apoptosis, we analyzed whether pUL46 of PrV is involved in signaling events impairing the integrity of the nuclear envelope. We show here that PrV pUL46 is able to induce phosphorylation of ERK1/2 and, thus, expression of ERK1/2 target genes but fails to activate the PI3K-Akt pathway. Deletion of UL46 from PrV-ΔUL34Pass and PrV-ΔUL31Pass or replacement by wild-type UL46 resulted in enhanced nuclear envelope breakdown, indicating that the mutations in pUL46 may limit the extent of NEBD. Thus, although pUL46 induces ERK1/2 phosphorylation, controlling the integrity of the nuclear envelope is independent of the ERK1/2 signaling pathway. IMPORTANCE Herpesvirus nucleocapsids can leave the nucleus by regulated, vesicle-mediated transport through the nuclear envelope, designated nuclear egress, or by inducing nuclear envelope breakdown (NEBD). The viral proteins involved in NEBD are unknown. We show here that the pseudorabies virus tegument protein pUL46 induces the

  20. Integrated treatment and handling of highly activated components from nuclear facilities

    SciTech Connect

    Schneider, K.A.; Kiolbassa, A.; Rose, K.A.; Raymont, J.M. Jr.

    1993-12-31

    A complete Underwater Treatment System (UTS) is described for activated/contaminated components of various origins in the nuclear industry. The system comprises different kinds of cutting/compacting equipment: the USC (Underwater Shear/compactor), the SCS (Stellite Corner Shear), the VLS (Velocity Limiter Shear) and the LCS (Light Crusher Shear). Transfer and loading equipment, the STB (Shielded Transfer Bell) provides safe and economic loading of containers with cut components. Operating experience and performance data are presented.

  1. Activation of the human mitochondrial transcription factor A gene by nuclear respiratory factors: a potential regulatory link between nuclear and mitochondrial gene expression in organelle biogenesis.

    PubMed Central

    Virbasius, J V; Scarpulla, R C

    1994-01-01

    Mitochondrial transcription factor A (mtTFA), the product of a nuclear gene, stimulates transcription from the two divergent mitochondrial promoters and is likely the principal activator of mitochondrial gene expression in vertebrates. Here we establish that the proximal promoter of the human mtTFA gene is highly dependent upon recognition sites for the nuclear respiratory factors, NRF-1 and NRF-2, for activity. These factors have been previously implicated in the activation of numerous nuclear genes that contribute to mitochondrial respiratory function. The affinity-purified factors from HeLa cells specifically bind to the mtTFA NRF-1 and NRF-2 sites through guanine nucleotide contacts that are characteristic for each site. Mutations in these contacts eliminate NRF-1 and NRF-2 binding and also dramatically reduce promoter activity in transfected cells. Although both factors contribute, NRF-1 binding appears to be the major determinant of promoter function. This dependence on NRF-1 activation is confirmed by in vitro transcription using highly purified recombinant proteins that display the same binding specificities as the HeLa cell factors. The activation of the mtTFA promoter by both NRF-1 and NRF-2 therefore provides a link between the expression of nuclear and mitochondrial genes and suggests a mechanism for their coordinate regulation during organelle biogenesis. Images PMID:8108407

  2. Src subfamily kinases regulate nuclear export and degradation of transcription factor Nrf2 to switch off Nrf2-mediated antioxidant activation of cytoprotective gene expression.

    PubMed

    Niture, Suryakant K; Jain, Abhinav K; Shelton, Phillip M; Jaiswal, Anil K

    2011-08-19

    Nrf2 (NF-E2-related factor 2) is a nuclear transcription factor that in response to chemical and radiation stress regulates coordinated induction of a battery of cytoprotective gene expressions leading to cellular protection. In this study, we investigated the role of Src kinases in the regulation of Nrf2 and downstream signaling. siRNA-mediated inhibition of Fyn, Src, Yes, and Fgr, but not Lyn, in mouse hepatoma Hepa-1 cells, led to nuclear accumulation of Nrf2 and up-regulation of Nrf2 downstream gene expression. Mouse embryonic fibroblasts with combined deficiency of Fyn/Src/Yes/Fgr supported results from siRNA. In addition, steady-state overexpression of Fyn, Src, and Yes phosphorylated Nrf2Tyr568 that triggered nuclear export and degradation of Nrf2 and down-regulation of Nrf2 downstream gene expression. Exposure of cells to antioxidant, oxidant, or UV radiation increased nuclear import of Fyn, Src, and Yes kinases, which phosphorylated Nrf2Tyr568 resulting in nuclear export and degradation of Nrf2. Further analysis revealed that stress-activated GSK3β acted upstream to the Src kinases and phosphorylated the Src kinases, leading to their nuclear localization and Nrf2 phosphorylation. The overexpression of Src kinases in Hepa-1 cells led to decreased Nrf2, increased apoptosis, and decreased cell survival. Mouse embryonic fibroblasts deficient in Src kinases showed nuclear accumulation of Nrf2, induction of Nrf2 and downstream gene expression, reduced apoptosis, and increased cell survival. The studies together demonstrate that Src kinases play a critical role in nuclear export and degradation of Nrf2, thereby providing a negative feedback mechanism to switch off Nrf2 activation and restore normal cellular homeostasis.

  3. Activation analysis study on Li-ion batteries for nuclear forensic applications

    NASA Astrophysics Data System (ADS)

    Johnson, Erik B.; Whitney, Chad; Holbert, Keith E.; Zhang, Taipeng; Stannard, Tyler; Christie, Anthony; Harper, Peter; Anderson, Blake; Christian, James F.

    2015-06-01

    The nuclear materials environment has been increasing significantly in complexity over the past couple of decades. The prevention of attacks from nuclear weapons is becoming more difficult, and nuclear forensics is a deterrent by providing detailed information on any type of nuclear event for proper attribution. One component of the nuclear forensic analysis is a measurement of the neutron spectrum. As an example, the neutron component provides information on the composition of the weapons, whether boosting is involved or the mechanisms used in creating a supercritical state. As 6Li has a large cross-section for thermal neutrons, the lithium battery is a primary candidate for assessing the neutron spectrum after detonation. The absorption process for 6Li yields tritium, which can be measured at a later point after the nuclear event, as long as the battery can be processed in a manner to successfully extract the tritium content. In addition, measuring the activated constituents after exposure provides a means to reconstruct the incident neutron spectrum. The battery consists of a spiral or folded layers of material that have unique, energy dependent interactions associated with the incident neutron flux. A detailed analysis on the batteries included a pre-irradiated mass spectrometry analysis to be used as input for neutron spectrum reconstruction. A set of batteries were exposed to a hard neutron spectrum delivered by the University of Massachusetts, Lowell research reactor Fast Neutron Irradiator (FNI). The gamma spectra were measured from the batteries within a few days and within a week after the exposure to obtain sufficient data on the activated materials in the batteries. The activity was calculated for a number of select isotopes, indicating the number of associated neutron interactions. The results from tritium extraction are marginal. A measurable increase in detected particles (gammas and betas) below 50 keV not self-attenuated by the battery was observed

  4. The Importance of Landscape Elements for Bat Activity and Species Richness in Agricultural Areas

    PubMed Central

    Heim, Olga; Treitler, Julia T.; Tschapka, Marco; Knörnschild, Mirjam; Jung, Kirsten

    2015-01-01

    Landscape heterogeneity is regarded as a key factor for maintaining biodiversity and ecosystem function in production landscapes. We investigated whether grassland sites at close vicinity to forested areas are more frequently used by bats. Considering that bats are important consumers of herbivorous insects, including agricultural pest, this is important for sustainable land management. Bat activity and species richness were assessed using repeated monitoring from May to September in 2010 with acoustic monitoring surveys on 50 grassland sites in the Biosphere Reserve Schorfheide-Chorin (North-East Germany). Using spatial analysis (GIS), we measured the closest distance of each grassland site to potentially connecting landscape elements (e.g., trees, linear vegetation, groves, running and standing water). In addition, we assessed the distance to and the percent land cover of forest remnants and urban areas in a 200 m buffer around the recording sites to address differences in the local landscape setting. Species richness and bat activity increased significantly with higher forest land cover in the 200 m buffer and at smaller distance to forested areas. Moreover, species richness increased in proximity to tree groves. Larger amount of forest land cover and smaller distance to forest also resulted in a higher activity of bats on grassland sites in the beginning of the year during May, June and July. Landscape elements near grassland sites also influenced species composition of bats and species richness of functional groups (open, edge and narrow space foragers). Our results highlight the importance of forested areas, and suggest that agricultural grasslands that are closer to forest remnants might be better buffered against outbreaks of agricultural pest insects due to higher species richness and higher bat activity. Furthermore, our data reveals that even for highly mobile species such as bats, a very dense network of connecting elements within the landscape is

  5. The Importance of Landscape Elements for Bat Activity and Species Richness in Agricultural Areas.

    PubMed

    Heim, Olga; Treitler, Julia T; Tschapka, Marco; Knörnschild, Mirjam; Jung, Kirsten

    2015-01-01

    Landscape heterogeneity is regarded as a key factor for maintaining biodiversity and ecosystem function in production landscapes. We investigated whether grassland sites at close vicinity to forested areas are more frequently used by bats. Considering that bats are important consumers of herbivorous insects, including agricultural pest, this is important for sustainable land management. Bat activity and species richness were assessed using repeated monitoring from May to September in 2010 with acoustic monitoring surveys on 50 grassland sites in the Biosphere Reserve Schorfheide-Chorin (North-East Germany). Using spatial analysis (GIS), we measured the closest distance of each grassland site to potentially connecting landscape elements (e.g., trees, linear vegetation, groves, running and standing water). In addition, we assessed the distance to and the percent land cover of forest remnants and urban areas in a 200 m buffer around the recording sites to address differences in the local landscape setting. Species richness and bat activity increased significantly with higher forest land cover in the 200 m buffer and at smaller distance to forested areas. Moreover, species richness increased in proximity to tree groves. Larger amount of forest land cover and smaller distance to forest also resulted in a higher activity of bats on grassland sites in the beginning of the year during May, June and July. Landscape elements near grassland sites also influenced species composition of bats and species richness of functional groups (open, edge and narrow space foragers). Our results highlight the importance of forested areas, and suggest that agricultural grasslands that are closer to forest remnants might be better buffered against outbreaks of agricultural pest insects due to higher species richness and higher bat activity. Furthermore, our data reveals that even for highly mobile species such as bats, a very dense network of connecting elements within the landscape is

  6. UNCOVERING THE DEEPLY EMBEDDED ACTIVE GALACTIC NUCLEUS ACTIVITY IN THE NUCLEAR REGIONS OF THE INTERACTING GALAXY Arp 299

    SciTech Connect

    Alonso-Herrero, A.; Roche, P. F.; Esquej, P.; Colina, L.; González-Martín, O.; Ramos Almeida, C.; Asensio Ramos, A.; Rodríguez Espinosa, J. M.; Alvarez, C.; Pereira-Santaella, M.; Levenson, N. A.; Packham, C.; Mason, R. E.; Aretxaga, I.; Díaz-Santos, T.; Perlman, E.; Telesco, C. M.

    2013-12-10

    We present mid-infrared (MIR) 8-13 μm spectroscopy of the nuclear regions of the interacting galaxy Arp 299 (IC 694+NGC 3690) obtained with CanariCam (CC) on the 10.4 m Gran Telescopio Canarias (GTC). The high angular resolution (∼0.''3-0.''6) of the data allows us to probe nuclear physical scales between 60 and 120 pc, which is a factor of 10 improvement over previous MIR spectroscopic observations of this system. The GTC/CC spectroscopy displays evidence of deeply embedded active galactic nucleus (AGN) activity in both nuclei. The GTC/CC nuclear spectrum of NGC 3690/Arp 299-B1 can be explained as emission from AGN-heated dust in a clumpy torus with both a high covering factor and high extinction along the line of sight. The estimated bolometric luminosity of the AGN in NGC 3690 is 3.2 ± 0.6 × 10{sup 44} erg s{sup –1}. The nuclear GTC/CC spectrum of IC 694/Arp 299-A shows 11.3 μm polycyclic aromatic hydrocarbon emission stemming from a deeply embedded (A{sub V} ∼ 24 mag) region of less than 120 pc in size. There is also a continuum-emitting dust component. If associated with the putative AGN in IC 694, we estimate that it would be approximately five times less luminous than the AGN in NGC 3690. The presence of dual AGN activity makes Arp 299 a good example to study such phenomena in the early coalescence phase of interacting galaxies.

  7. Acetylation directs survivin nuclear localization to repress STAT3 oncogenic activity.

    PubMed

    Wang, Haijuan; Holloway, Michael P; Ma, Li; Cooper, Zachary A; Riolo, Matthew; Samkari, Ayman; Elenitoba-Johnson, Kojo S J; Chin, Y Eugene; Altura, Rachel A

    2010-11-12

    The multiple functions of the oncofetal protein survivin are dependent on its selective expression patterns within immunochemically distinct subcellular pools. The mechanism by which survivin localizes to these compartments, however, is only partly understood. Here we show that nuclear accumulation of survivin is promoted by CREB-binding protein (CBP)-dependent acetylation on lysine 129 (129K, Lys-129). We demonstrate a mechanism by which survivin acetylation at this position results in its homodimerization, while deacetylation promotes the formation of survivin monomers that heterodimerize with CRM1 and facilitate its nuclear export. Using proteomic analysis, we identified the oncogenic transcription factor STAT3 as a binding partner of nuclear survivin. We show that acetylated survivin binds to the N-terminal transcriptional activation domain of the STAT3 dimer and represses STAT3 transactivation of target gene promoters. Using multiplex PCR and DNA sequencing, we identified a single-nucleotide polymorphism (A → G) at Lys-129 that exists as a homozygous mutation in a neuroblastoma cell line and corresponds with a defect in survivin nuclear localization. Our results demonstrate that the dynamic equilibrium between survivin acetylation and deacetylation at amino acid 129 determines its interaction with CRM1, its subsequent subcellular localization, and its ability to inhibit STAT3 transactivation, providing a potential route for therapeutic intervention in STAT3-dependent tumors.

  8. O-GlcNAc-glycosylation of {beta}-catenin regulates its nuclear localization and transcriptional activity

    SciTech Connect

    Sayat, Ria; Leber, Brian; Grubac, Vanja; Wiltshire, Lesley; Persad, Sujata

    2008-09-10

    {beta}-catenin plays a role in intracellular adhesion and regulating gene expression. The latter role is associated with its oncogenic properties. Phosphorylation of {beta}-catenin controls its intracellular expression but mechanism/s that regulates the nuclear localization of {beta}-catenin is unknown. We demonstrate that O-GlcNAc glycosylation (O-GlcNAcylation) of {beta}-catenin negatively regulates its levels in the nucleus. We show that normal prostate cells (PNT1A) have significantly higher amounts of O-GlcNAcylated {beta}-catenin compared to prostate cancer (CaP) cells. The total nuclear levels of {beta}-catenin are higher in the CaP cells than PNT1A but only a minimal fraction of the nuclear {beta}-catenin in the CaP cells are O-GlcNAcylated. Increasing the levels of O-GlcNAcylated {beta}-catenin in the CaP cells with PUGNAc (O- (2-acetamido-2-deoxy-D-gluco-pyranosylidene) amino-N-phenylcarbamate) treatment is associated with a progressive decrease in the levels of {beta}-catenin in the nucleus. TOPFlash reporter assay and mRNA expressions of {beta}-catenin's target genes indicate that O-GlcNAcylation of {beta}-catenin results in a decrease in its transcriptional activity. We define a novel modification of {beta}-catenin that regulates its nuclear localization and transcriptional function.

  9. [Social support and physical activity in adolescents from public schools: the importance of family and friends].

    PubMed

    Prado, Crisley Vanessa; Lima, Alex Vieira; Fermino, Rogério César; Añez, Ciro Romelio Rodriguez; Reis, Rodrigo Siqueira

    2014-04-01

    The aim of this study was to verify the association between different types and sources of social support and physical activity among adolescents from Curitiba, Paraná State, Brazil. A school-based survey was conducted with a representative sample of adolescents from public schools (n = 1,469). Multiple regression models were used to test the association between weekly frequency and sources of social support from family and friends and weekly frequency of physical activity. Among boys, frequent company of family (PR: 2.88; 95%CI: 2.00-4.13) and friends (PR: 5.46; 95%CI: 2.33-12.78) and positive reinforcement from friends (PR: 1.81; 95%CI: 1.18-2.77) were positively associated with physical activity. Sporadic invitation by the family was negatively associated with physical activity (PR: 0.66; 95%CI: 0.46-1.14). For girls, frequent company of family (PR: 3.39; 95%CI: 1.49-7.69) and friends (PR: 4.06; 95%CI: 2.22-7.45) increased the likelihood of physical activity. Company of friends was the most important type of social support for physical activity among these adolescents.

  10. Mitochondria play an important role in the cell proliferation suppressing activity of berberine

    PubMed Central

    Yan, Xiao-Jin; Yu, Xuan; Wang, Xin-Pei; Jiang, Jing-Fei; Yuan, Zhi-Yi; Lu, Xi; Lei, Fan; Xing, Dong-Ming

    2017-01-01

    After being studied for approximately a century, berberine (BBR) has been found to act on various targets and pathways. A great challenge in the pharmacological analysis of BBR at present is to identify which target(s) plays a decisive role. In the study described herein, a rescue experiment was designed to show the important role of mitochondria in BBR activity. A toxic dose of BBR was applied to inhibit cell proliferation and mitochondrial activity, then α-ketobutyrate (AKB), an analogue of pyruvate that serves only as an electron receptor of NADH, was proven to partially restore cell proliferation. However, mitochondrial morphology damage and TCA cycle suppression were not recovered by AKB. As the AKB just help to regenerate NAD+, which is make up for part function of mitochondrial, the recovered cell proliferation stands for the contribution of mitochondria to the activity of BBR. Our results also indicate that BBR suppresses tumour growth and reduces energy charge and mitochondrial DNA (mtDNA) copy number in a HepG2 xenograft model. In summary, our study suggests that mitochondria play an important role in BBR activity regarding tumour cell proliferation and metabolism. PMID:28181523

  11. Induction of apoptosis by ribosome inactivating proteins: importance of N-glycosidase activity.

    PubMed

    Das, Mrinal Kumar; Sharma, Radhey Shyam; Mishra, Vandana

    2012-03-01

    Apoptotic cell death is a fundamental process in the development and physiological homeostasis of multicellular organisms. It is associated with control of cell numbers in tissues and organs during development, with cell turnover, and with response to infection. Molecules that trigger this process in continuously proliferating cancer cells can be used as chemotherapeutic agents. Ribosome inactivating proteins (RIPs) that inhibit translation in a cell by depurinating (N-glycosidase activity) the 28S rRNA are known to serve as apoptosis inducers. However, the role of depurination activity of the RIPs in apoptosis induction is still controversial. Presently, there are three different hypotheses which propose that depurination is: (1) essential, (2) essential but not the sole factor, or (3) not essential for apoptosis induction. This article reviews various experimental outcomes on the importance of N-glycosidase activity of RIPs in the induction of apoptosis.

  12. Overview of a FPGA-based nuclear instrumentation dedicated to primary activity measurements.

    PubMed

    Bobin, C; Bouchard, J; Pierre, S; Thiam, C

    2012-09-01

    In National Metrology Institutes like LNE-LNHB, renewal and improvement of the instrumentation is an important task. Nowadays, the current trend is to adopt digital boards, which present numerous advantages over the standard electronics. The feasibility of an on-line fulfillment of nuclear-instrumentation functionalities using a commercial FPGA-based (Field-Programmable Gate Array) board has been validated in the case of TDCR primary measurements (Triple to Double Coincidence Ratio method based on liquid scintillation). The new applications presented in this paper have been included to allow either an on-line processing of the information or a raw-data acquisition for an off-line treatment. Developed as a complementary tool for TDCR counting, a time-to-digital converter specifically designed for this technique has been added. In addition, the description is given of a spectrometry channel based on the connection between conventional shaping amplifiers and the analog-to-digital converter (ADC) input available on the same digital board. First results are presented in the case of α- and γ-counting related to, respectively, the defined solid angle and well-type NaI(Tl) primary activity techniques. The combination of two different channels (liquid scintillation and γ-spectrometry) implementing the live-time anticoincidence processing is also described for the application of the 4πβ-γ coincidence method. The need for an optimized coupling between the analog chain and the ADC stage is emphasized. The straight processing of the signals delivered by the preamplifier connected to a HPGe detector is also presented along with the first development of digital filtering.

  13. Probing the nuclear and circumnuclear activity of NGC 1365 in the infrared

    NASA Astrophysics Data System (ADS)

    Alonso-Herrero, A.; Sánchez-Portal, M.; Ramos Almeida, C.; Pereira-Santaella, M.; Esquej, P.; García-Burillo, S.; Castillo, M.; González-Martín, O.; Levenson, N.; Hatziminaoglou, E.; Acosta-Pulido, J. A.; González-Serrano, J. I.; Pović, M.; Packham, C.; Pérez-García, A. M.

    2012-09-01

    We present new far-infrared (70-500 μm) Herschel Photodetector Array Camera and Spectrometer (PACS) and Spectral and Photometric Imaging Receiver (SPIRE) imaging observations as well as new mid-IR Gemini/Thermal-Region Camera Spectrograph imaging (8.7 and 18.3 μm) and spectroscopy of the inner Lindblad resonance (ILR) region (R < 2.5 kpc) of the spiral galaxy NGC 1365. We complemented these observations with archival Spitzer imaging and spectral mapping observations. The ILR region of NGC 1365 contains a Seyfert 1.5 nucleus and a ring of star formation with an approximate diameter of 2 kpc. The strong star formation activity in the ring is resolved by the Herschel/PACS imaging data, as well as by the Spitzer 24 μm continuum emission, [Ne II] 12.81 μm line emission, and 6.2 and 11.3 μm PAH emission. The active galactic nucleus (AGN) is the brightest source in the central regions up to λ ˜ 24 μm, but it becomes increasingly fainter in the far-infrared when compared to the emission originating in the infrared clusters (or groups of them) located in the ring. We modelled the AGN unresolved infrared emission with the clumpy torus models and estimated that the AGN contributes only to a small fraction (˜5 per cent) of the infrared emission produced in the inner ˜5 kpc. We fitted the non-AGN 24-500 μm spectral energy distribution of the ILR region and found that the dust temperatures and mass are similar to those of other nuclear and circumnuclear starburst regions. Finally we showed that within the ILR region of NGC 1365, most of the ongoing star formation activity is taking place in dusty regions as probed by the 24 μm emission. Herschel is an ESA space observatory with science instruments provided by European-led Principal Investigator consortia and with important participation from NASA.Augusto González Linares Senior Research Fellow.

  14. Moraxella catarrhalis induces mast cell activation and nuclear factor kappa B-dependent cytokine synthesis.

    PubMed

    Krishnaswamy, G; Martin, R; Walker, E; Li, C; Hossler, F; Hall, K; Chi, D S

    2003-01-01

    Human mast cells are often found perivascularly and at mucosal sites and may play crucial roles in the inflammatory response. Recent studies have suggested a prominent role for mast cells in host defense. In this study, we analyzed the effects of a common airway pathogen, Moraxella catarrhalis and a commensal bacterium, Neiserria cinerea, on activation of human mast cells. Human mast cell leukemia cells (HMC-1) were activated with either phorbol myristate acetate (PMA) and calcium ionophore or with varying concentrations of heat-killed suspensions of bacteria. Supernatants were assayed for the cytokines interleukin-4 (IL-4), granulocyte macrophage colony stimulating factor (GM-CSF), IL-6, IL-8, IL-13 and monocyte chemotactic protein-1 (MCP-1). Nuclear proteins were isolated and assayed by electrophoretic mobility shift assay (EMSA) for nuclear factor kappaB (NF-kappaB) nuclear binding activity. In some experiments, NF-kappaB inhibitor, Bay-11 was added to determine functional significance. Both M. catarrhalis and N. cinerea induced mast cell activation and selective secretion of two key inflammatory cytokines, IL-6 and MCP-1. This was accompanied by NF-kappaB activation. Neither spun bacterial supernatants nor bacterial lipopolysaccharide induced cytokine secretion, suggesting need for direct bacterial contact with mast cells. Scanning electron microscopy revealed active aggregation of bacteria over mast cell surfaces. The NF-kappaB inhibitor, Bay-11, inhibited expression of MCP-1. These findings suggest the possibility of direct interactions between human mast cells and common bacteria and provide evidence for a novel role for human mast cells in innate immunity.

  15. Cooperative effects of hepatitis B virus and TNF may play important roles in the activation of metabolic pathways through the activation of NF-κB

    PubMed Central

    Wu, Shuang; Kanda, Tatsuo; Nakamoto, Shingo; Jiang, Xia; Nakamura, Masato; Sasaki, Reina; Haga, Yuki; Shirasawa, Hiroshi; Yokosuka, Osamu

    2016-01-01

    Elevated levels of inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β are often observed in the sera of hepatitis B virus (HBV)-infected patients. It is well known that these cytokines activate nuclear factor-κB (NF-κB)-signaling, and are associated with endoplasmic reticulum (ER) stress. We investigated whether HBV or HBV X protein (HBx) enhanced the activation of NF-κB in the presence of TNF and/or IL-1β, and their effects on the expression of metabolic pathway-associated genes. We examined whether HBV or HBx enhanced cytokine-induced activation of NF-κB in hepatocytes, using a reporter assay, in the presence or absence of TNF and/or IL-1β. The expression of insulin-like growth factor binding protein 1 (IGFBP1), one of the NF-κB target genes was also examined. The expression of metabolic pathway-associated genes in HepG2 and HepG2.2.15 cells in the presence or absence of TNF was evaluated by RT-qPCR. Human hepatocytes expressed TNF receptors and IL-1 receptors. NF-κB was activated by cooperation between HBx and TNF in human hepatocytes. We observed IGFBP1 expression in HBV infection and that a number of metabolic pathway-associated genes were upregulated in HepG2.2.15 cells, compared with HepG2 cells with or without TNF treatment. We observed the cooperative effects of HBV and TNF which enhanced the activation of NF-κB as well as upregulated the expression of metabolic pathway-associated genes in hepatocytes. These effects may be important in the development of HBV-associated metabolic syndrome. PMID:27315566

  16. Cooperative effects of hepatitis B virus and TNF may play important roles in the activation of metabolic pathways through the activation of NF-κB.

    PubMed

    Wu, Shuang; Kanda, Tatsuo; Nakamoto, Shingo; Jiang, Xia; Nakamura, Masato; Sasaki, Reina; Haga, Yuki; Shirasawa, Hiroshi; Yokosuka, Osamu

    2016-08-01

    Elevated levels of inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β are often observed in the sera of hepatitis B virus (HBV)-infected patients. It is well known that these cytokines activate nuclear factor-κB (NF-κB)-signaling, and are associated with endoplasmic reticulum (ER) stress. We investigated whether HBV or HBV X protein (HBx) enhanced the activation of NF-κB in the presence of TNF and/or IL-1β, and their effects on the expression of metabolic pathway‑associated genes. We examined whether HBV or HBx enhanced cytokine-induced activation of NF-κB in hepatocytes, using a reporter assay, in the presence or absence of TNF and/or IL-1β. The expression of insulin-like growth factor binding protein 1 (IGFBP1), one of the NF-κB target genes was also examined. The expression of metabolic pathway-associated genes in HepG2 and HepG2.2.15 cells in the presence or absence of TNF was evaluated by RT-qPCR. Human hepatocytes expressed TNF receptors and IL-1 receptors. NF-κB was activated by cooperation between HBx and TNF in human hepatocytes. We observed IGFBP1 expression in HBV infection and that a number of metabolic pathway-associated genes were upregulated in HepG2.2.15 cells, compared with HepG2 cells with or without TNF treatment. We observed the cooperative effects of HBV and TNF which enhanced the activation of NF-κB as well as upregulated the expression of metabolic pathway-associated genes in hepatocytes. These effects may be important in the development of HBV-associated metabolic syndrome.

  17. The human papillomavirus (HPV) E6 oncoproteins promotes nuclear localization of active caspase 8

    SciTech Connect

    Manzo-Merino, Joaquin; Lizano, Marcela

    2014-02-15

    The HPV-16 E6 and E6{sup ⁎} proteins have been shown previously to be capable of regulating caspase 8 activity. We now show that the capacity of E6 to interact with caspase 8 is common to diverse HPV types, being also seen with HPV-11 E6, HPV-18 E6 and HPV-18 E6{sup ⁎}. Unlike most E6-interacting partners, caspase 8 does not appear to be a major proteasomal target of E6, but instead E6 appears able to stimulate caspase 8 activation, without affecting the overall apoptotic activity. This would appear to be mediated in part by the ability of the HPV E6 oncoproteins to recruit active caspase 8 to the nucleus. - Highlights: • Multiple HPV E6 oncoproteins interact with the caspase 8 DED domain. • HPV E6 stimulates activation of caspase 8. • HPV E6 promotes nuclear accumulation of caspase 8.

  18. Studies of Neutron and Proton Nuclear Activation in Low-Earth Orbit 2

    NASA Technical Reports Server (NTRS)

    Laird, C. E.

    1983-01-01

    The study of neutron and proton nuclear activation in low-Earth orbit reported in NASA CR-162051 has been continued with increasing emphasis given to primary and secondary neutron activation. The previously reported activation due to protons has been modified to include: (1) flux attenuation caused by all inelastic reactions; (2) the modification of the proton flux distribution caused by sample covering material; and (3) the activation of the sample as a function of the distance into the sample from the surface of incidence. A method has been developed for including the effects on the activation of the finite width and length of the samples. The reactant product spectra produced by proton-induced reactions has been studied. Cross sections needed for neutron induced reactions leading to long-lived (half-life 1 day) radioisotopes have been identified and, in some cases, compiled.

  19. The importance of thiopurine methyltransferase activity for the use of azathioprine in transplant recipients.

    PubMed

    Chocair, P R; Duley, J A; Simmonds, H A; Cameron, J S

    1992-05-01

    immunosuppression apparently occurred, based on low catabolism of the drug. We conclude that total intolerance to azathioprine is rare and usually appears in patients with very low TPMT activities. Our results also suggest that the wide range of TPMT activity may be an important factor in determining long-term graft survival in azathioprine-treated patients; those with high activity might benefit from doses near the upper limit generally recommended.

  20. A new perspective on the radio active zone at the Galactic center - feedback from nuclear activities

    NASA Astrophysics Data System (ADS)

    Zhao, J.-H.; Morris, M. R.; Goss, W. M.

    2014-05-01

    Based on our deep image of Sgr A using broadband data observed with the VLA† at 6 cm, we present a new perspective of the radio bright zone at the Galactic center. We further show the radio detection of the X-ray Cannonball, a candidate neutron star associated with the Galactic center SNR Sgr A East. The radio image is compared with the Chandra X-ray image to show the detailed structure of the radio counterparts of the bipolar X-ray lobes. The bipolar lobes are likely produced by the winds from the activities within Sgr A West, which could be collimated by the inertia of gas in the CND, or by the momentum driving of Sgr A*; and the poloidal magnetic fields likely play an important role in the collimation. The less-collimated SE lobe, in comparison to the NW one, is perhaps due to the fact that the Sgr A East SN might have locally reconfigured the magnetic field toward negative galactic latitudes. In agreement with the X-ray observations, the time-scale of ˜1 × 104 yr estimated for the outermost radio ring appears to be comparable to the inferred age of the Sgr A East SNR.

  1. EGF Regulation of Nuclear Co-Activator AIB1 Function in Breast Cancer

    DTIC Science & Technology

    2005-04-01

    AD Award Number: DAMDl7-02-1-0394 TITLE: EGF Regulation of Nuclear Co-Activator AIBI Function in Breast Cancer PRINCIPAL INVESTIGATOR: Annabell S. Oh...Activator AIBI Function in DAMD17-02-1-0394 Breast Cancer 6. AUTHOR(S) Annabell S. Oh 7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) 8. PERFORMING...Factor-i Signaling in Human Breast Cancer. Doctoral dissertation of Annabell S. Oh, B.S. from the Department 14 of Tumor Biology, Georgetown

  2. International Nuclear Safeguards at Sandia

    SciTech Connect

    Sternat, Matthew R.

    2015-02-01

    As global nuclear energy expands, assuring peaceful uses of nuclear technology becomes increasingly important. In addition to complying with international nuclear safeguards, a responsible nuclear energy program promotes a corresponding safeguards culture. Establishment of transparent peaceful uses of nuclear technologies starts with cooperative international engagements and safeguards systems. Developing states investing in nuclear energy must assure the international community of their longterm commitment to safeguards, safety, and security (3S) of nuclear materials and technologies. Cultivating a safeguards culture starts in the initial phases of infrastructure planning and must be integrated into the process of developing a responsible nuclear energy program. Sandia National Laboratories supports the implementation of safeguards culture through a variety of activities, including infrastructure development.

  3. Multiple active site residues are important for photochemical efficiency in the light-activated enzyme protochlorophyllide oxidoreductase (POR).

    PubMed

    Menon, Binuraj R K; Hardman, Samantha J O; Scrutton, Nigel S; Heyes, Derren J

    2016-08-01

    Protochlorophyllide oxidoreductase (POR) catalyzes the light-driven reduction of protochlorophyllide (Pchlide), an essential, regulatory step in chlorophyll biosynthesis. The unique requirement of the enzyme for light has provided the opportunity to investigate how light energy can be harnessed to power biological catalysis and enzyme dynamics. Excited state interactions between the Pchlide molecule and the protein are known to drive the subsequent reaction chemistry. However, the structural features of POR and active site residues that are important for photochemistry and catalysis are currently unknown, because there is no crystal structure for POR. Here, we have used static and time-resolved spectroscopic measurements of a number of active site variants to study the role of a number of residues, which are located in the proposed NADPH/Pchlide binding site based on previous homology models, in the reaction mechanism of POR. Our findings, which are interpreted in the context of a new improved structural model, have identified several residues that are predicted to interact with the coenzyme or substrate. Several of the POR variants have a profound effect on the photochemistry, suggesting that multiple residues are important in stabilizing the excited state required for catalysis. Our work offers insight into how the POR active site geometry is finely tuned by multiple active site residues to support enzyme-mediated photochemistry and reduction of Pchlide, both of which are crucial to the existence of life on Earth.

  4. Daily variation of constitutively activated nuclear factor kappa B (NFKB) in rat pineal gland.

    PubMed

    Cecon, Erika; Fernandes, Pedro A; Pinato, Luciana; Ferreira, Zulma S; Markus, Regina P

    2010-01-01

    In mammals, the production of melatonin by the pineal gland is mainly controlled by the suprachiasmatic nuclei (SCN), the master clock of the circadian system. We have previously shown that agents involved in inflammatory responses, such as cytokines and corticosterone, modulate pineal melatonin synthesis. The nuclear transcription factor NFKB, detected by our group in the rat pineal gland, modulates this effect. Here, we evaluated a putative constitutive role for the pineal gland NFKB pathway. Male rats were kept under 12 h:12 h light-dark (LD) cycle or under constant darkness (DD) condition. Nuclear NFKB was quantified by electrophoretic mobility shift assay on pineal glands obtained from animals killed throughout the day at different times. Nuclear content of NFKB presented a daily rhythm only in LD-entrained animals. During the light phase, the amount of NFKB increased continuously, and a sharp drop occurred when lights were turned off. Animals maintained in a constant light environment until ZT 18 showed diurnal levels of nuclear NFKB at ZT15 and ZT18. Propranolol (20 mg/kg, i.p., ZT 11) treatment, which inhibits nocturnal sympathetic input, impaired nocturnal decrease of NFKB only at ZT18. A similar effect was observed in free-running animals, which secreted less nocturnal melatonin. Because melatonin reduces constitutive NFKB activation in cultured pineal glands, we propose that this indolamine regulates this transcription factor pathway in the rat pineal gland, but not at the LD transition. The controversial results regarding the inhibition of pineal function by constant light or blocking sympathetic neurotransmission are discussed according to the hypothesis that the prompt effect of lights-off is not mediated by noradrenaline, which otherwise contributes to maintaining low levels of nuclear NFKB at night. In summary, we report here a novel transcription factor in the pineal gland, which exhibits a constitutive rhythm dependent on environmental photic

  5. Importance of rostral ventrolateral medulla neurons in determining efferent sympathetic nerve activity and blood pressure.

    PubMed

    Kumagai, Hiroo; Oshima, Naoki; Matsuura, Tomokazu; Iigaya, Kamon; Imai, Masaki; Onimaru, Hiroshi; Sakata, Katsufumi; Osaka, Motohisa; Onami, Toshiko; Takimoto, Chie; Kamayachi, Tadashi; Itoh, Hiroshi; Saruta, Takao

    2012-02-01

    Accentuated sympathetic nerve activity (SNA) is a risk factor for cardiovascular events. In this review, we investigate our working hypothesis that potentiated activity of neurons in the rostral ventrolateral medulla (RVLM) is the primary cause of experimental and essential hypertension. Over the past decade, we have examined how RVLM neurons regulate peripheral SNA, how the sympathetic and renin-angiotensin systems are correlated and how the sympathetic system can be suppressed to prevent cardiovascular events in patients. Based on results of whole-cell patch-clamp studies, we report that angiotensin II (Ang II) potentiated the activity of RVLM neurons, a sympathetic nervous center, whereas Ang II receptor blocker (ARB) reduced RVLM activities. Our optical imaging demonstrated that a longitudinal rostrocaudal column, including the RVLM and the caudal end of ventrolateral medulla, acts as a sympathetic center. By organizing and analyzing these data, we hope to develop therapies for reducing SNA in our patients. Recently, 2-year depressor effects were obtained by a single procedure of renal nerve ablation in patients with essential hypertension. The ablation injured not only the efferent renal sympathetic nerves but also the afferent renal nerves and led to reduced activities of the hypothalamus, RVLM neurons and efferent systemic sympathetic nerves. These clinical results stress the importance of the RVLM neurons in blood pressure regulation. We expect renal nerve ablation to be an effective treatment for congestive heart failure and chronic kidney disease, such as diabetic nephropathy.

  6. The Nuclear Hormone Receptor Peroxisome Proliferator-Activated Receptor β/δ Potentiates Cell Chemotactism, Polarization, and Migration▿ †

    PubMed Central

    Tan, Nguan Soon; Icre, Guillaume; Montagner, Alexandra; Heggeler, Béatrice Bordier-ten; Wahli, Walter; Michalik, Liliane

    2007-01-01

    After an injury, keratinocytes acquire the plasticity necessary for the reepithelialization of the wound. Here, we identify a novel pathway by which a nuclear hormone receptor, until now better known for its metabolic functions, potentiates cell migration. We show that peroxisome proliferator-activated receptor β/δ (PPARβ/δ) enhances two phosphatidylinositol 3-kinase-dependent pathways, namely, the Akt and the Rho-GTPase pathways. This PPARβ/δ activity amplifies the response of keratinocytes to a chemotactic signal, promotes integrin recycling and remodeling of the actin cytoskeleton, and thereby favors cell migration. Using three-dimensional wound reconstructions, we demonstrate that these defects have a strong impact on in vivo skin healing, since PPARβ/δ−/− mice show an unexpected and rare epithelialization phenotype. Our findings demonstrate that nuclear hormone receptors not only regulate intercellular communication at the organism level but also participate in cell responses to a chemotactic signal. The implications of our findings may be far-reaching, considering that the mechanisms described here are important in many physiological and pathological situations. PMID:17682064

  7. Domestic politics, citizen activism, and U. S. nuclear arms control policy

    SciTech Connect

    Knopf, J.W.

    1991-01-01

    The author seeks to ascertain whether and how citizens' movements concerning nuclear arms control and disarmament affect US arms control policy. The author employs a comparative case study methodology. He examines cases of the Eisenhower and Kennedy Administrations during the period of protest against nuclear testing, and the Reagan Administration during the nuclear weapons freeze campaign and the subsequent campaign for a comprehensive test ban. He hows there are four mechanisms through which public advocacy efforts can influence arms control policy, identifies the conditions under which each can be effective, and details the type of impact each mechanism has. Domestic activism interacts with broader public opinion in a way that creates electoral pressure; with elite-level debates in a way that removes a consensus behind presidential policy or changes the winning coalition in Congress; with bureaucratic politics, by generating ideas that have utility for some agents within the Executive; or with the public diplomacy of foreign governments, especially the Soviet Union. Citizens' movements had an impact on policy in each of the cases studied. The type and extent of impact, and the mechanisms involved in giving activism influence, are different for each case.

  8. Cognitive activities during adulthood are more important than education in building reserve.

    PubMed

    Reed, Bruce R; Dowling, Maritza; Tomaszewski Farias, Sarah; Sonnen, Joshua; Strauss, Milton; Schneider, Julie A; Bennett, David A; Mungas, Dan

    2011-07-01

    Cognitive reserve is thought to reflect life experiences. Which experiences contribute to reserve and their relative importance is not understood. Subjects were 652 autopsied cases from the Rush Memory and Aging Project and the Religious Orders Study. Reserve was defined as the residual variance of the regressions of cognitive factors on brain pathology and was captured in a latent variable that was regressed on potential determinants of reserve. Neuropathology variables included Alzheimer's disease markers, Lewy bodies, infarcts, microinfarcts, and brain weight. Cognition was measured with six cognitive domain scores. Determinants of reserve were socioeconomic status (SES), education, leisure cognitive activities at age 40 (CA40) and at study enrollment (CAbaseline) in late life. The four exogenous predictors of reserve were weakly to moderately inter-correlated. In a multivariate model, all except SES had statistically significant effects on Reserve, the strongest of which were CA40 (β = .31) and CAbaseline (β = .28). The Education effect was negative in the full model (β = -.25). Results suggest that leisure cognitive activities throughout adulthood are more important than education in determining reserve. Discrepancies between cognitive activity and education may be informative in estimating late life reserve.

  9. Prolonged interval between fusion and activation impairs embryonic development by inducing chromosome scattering and nuclear aneuploidy in pig somatic cell nuclear transfer.

    PubMed

    You, Jinyoung; Song, Kilyoung; Lee, Eunsong

    2010-01-01

    The aim of the present study was to examine the effect of various intervals between electrofusion and activation (FA interval) on the nuclear remodelling and development of somatic cell nuclear transfer (SCNT) embryos in pigs. Reconstructed oocytes were activated at 0 (simultaneous fusion and activation; SFA), 1, 2 and 3 h (delayed activation) after electrofusion; these groups were designated as DA1, DA2 and DA3, respectively. When oocyte nuclear status was examined at 0.5, 1, 2 and 3 h after electrofusion, the incidence of chromosome scattering was increased (P < 0.01) as the FA interval was extended (0.0%, 12.0%, 77.3% and 78.0%, respectively). Extending the FA interval led to an increase (P < 0.01) in the percentage of oocytes containing multiple (>or=3) pseudopronuclei (PPN) (0.0% of SFA; 5.3% of DA1; 21.7% of DA2; and 33.5% of DA3). The development of SCNT embryos to the blastocyst stage was decreased (P < 0.05) in DA2 (5.7%) and DA3 (5.0%) compared with SFA (18.1%) and DA1 (19.5%). Our results demonstrate that extending the FA interval impairs the development of SCNT pig embryos by inducing chromosome scattering and the formation of multiple PPN, which may result in increased nuclear aneuploidy.

  10. Nuclear Rocket Test Facility Decommissioning Including Controlled Explosive Demolition of a Neutron-Activated Shield Wall

    SciTech Connect

    Michael Kruzic

    2007-09-01

    Located in Area 25 of the Nevada Test Site, the Test Cell A Facility was used in the 1960s for the testing of nuclear rocket engines, as part of the Nuclear Rocket Development Program. The facility was decontaminated and decommissioned (D&D) in 2005 using the Streamlined Approach For Environmental Restoration (SAFER) process, under the Federal Facilities Agreement and Consent Order (FFACO). Utilities and process piping were verified void of contents, hazardous materials were removed, concrete with removable contamination decontaminated, large sections mechanically demolished, and the remaining five-foot, five-inch thick radiologically-activated reinforced concrete shield wall demolished using open-air controlled explosive demolition (CED). CED of the shield wall was closely monitored and resulted in no radiological exposure or atmospheric release.

  11. THE NUCLEAR INFRARED EMISSION OF LOW-LUMINOSITY ACTIVE GALACTIC NUCLEI

    SciTech Connect

    Mason, R. E.; Lopez-Rodriguez, E.; Packham, C.; Alonso-Herrero, A.; Elitzur, M.; Aretxaga, I.; Roche, P. F.; Oi, N.

    2012-07-15

    We present high-resolution mid-infrared (MIR) imaging, nuclear spectral energy distributions (SEDs), and archival Spitzer spectra for 22 low-luminosity active galactic nuclei (LLAGNs; L{sub bol} {approx}< 10{sup 42} erg s{sup -1}). Infrared (IR) observations may advance our understanding of the accretion flows in LLAGNs, the fate of the obscuring torus at low accretion rates, and, perhaps, the star formation histories of these objects. However, while comprehensively studied in higher-luminosity Seyferts and quasars, the nuclear IR properties of LLAGNs have not yet been well determined. We separate the present LLAGN sample into three categories depending on their Eddington ratio and radio emission, finding different IR characteristics for each class. (1) At the low-luminosity, low-Eddington-ratio (log L{sub bol}/L{sub Edd} < -4.6) end of the sample, we identify 'host-dominated' galaxies with strong polycyclic aromatic hydrocarbon bands that may indicate active (circum-)nuclear star formation. (2) Some very radio-loud objects are also present at these low Eddington ratios. The IR emission in these nuclei is dominated by synchrotron radiation, and some are likely to be unobscured type 2 AGNs that genuinely lack a broad-line region. (3) At higher Eddington ratios, strong, compact nuclear sources are visible in the MIR images. The nuclear SEDs of these galaxies are diverse; some resemble typical Seyfert nuclei, while others lack a well-defined MIR 'dust bump'. Strong silicate emission is present in many of these objects. We speculate that this, together with high ratios of silicate strength to hydrogen column density, could suggest optically thin dust and low dust-to-gas ratios, in accordance with model predictions that LLAGNs do not host a Seyfert-like obscuring torus. We anticipate that detailed modeling of the new data and SEDs in terms of accretion disk, jet, radiatively inefficient accretion flow, and torus components will provide further insights into the nuclear

  12. Nitric oxide modulates chromatin folding in human endothelial cells via protein phosphatase 2A activation and class II histone deacetylases nuclear shuttling.

    PubMed

    Illi, Barbara; Dello Russo, Claudio; Colussi, Claudia; Rosati, Jessica; Pallaoro, Michele; Spallotta, Francesco; Rotili, Dante; Valente, Sergio; Ragone, Gianluca; Martelli, Fabio; Biglioli, Paolo; Steinkuhler, Christian; Gallinari, Paola; Mai, Antonello; Capogrossi, Maurizio C; Gaetano, Carlo

    2008-01-04

    Nitric oxide (NO) modulates important endothelial cell (EC) functions and gene expression by a molecular mechanism which is still poorly characterized. Here we show that in human umbilical vein ECs (HUVECs) NO inhibited serum-induced histone acetylation and enhanced histone deacetylase (HDAC) activity. By immunofluorescence and Western blot analyses it was found that NO induced class II HDAC4 and 5 nuclear shuttling and that class II HDACs selective inhibitor MC1568 rescued serum-dependent histone acetylation above control level in NO-treated HUVECs. In contrast, class I HDACs inhibitor MS27-275 had no effect, indicating a specific role for class II HDACs in NO-dependent histone deacetylation. In addition, it was found that NO ability to induce HDAC4 and HDAC5 nuclear shuttling involved the activation of the protein phosphatase 2A (PP2A). In fact, HDAC4 nuclear translocation was impaired in ECs expressing small-t antigen and exposed to NO. Finally, in cells engineered to express a HDAC4-Flag fusion protein, NO induced the formation of a macromolecular complex including HDAC4, HDAC3, HDAC5, and an active PP2A. The present results show that NO-dependent PP2A activation plays a key role in class II HDACs nuclear translocation.

  13. Retinoids induce integrin-independent lymphocyte adhesion through RAR-α nuclear receptor activity

    SciTech Connect

    Whelan, Jarrett T.; Wang, Lei; Chen, Jianming; Metts, Meagan E.; Nasser, Taj A.; McGoldrick, Liam J.; Bridges, Lance C.

    2014-11-28

    Highlights: • Transcription and translation are required for retinoid-induced lymphocyte adhesion. • RAR activation is sufficient to induced lymphocyte cell adhesion. • Vitamin D derivatives inhibit RAR-prompted lymphocyte adhesion. • Adhesion occurs through a novel binding site within ADAM disintegrin domains. • RARα is a key nuclear receptor for retinoid-dependent lymphocyte cell adhesion. - Abstract: Oxidative metabolites of vitamin A, in particular all-trans-retinoic acid (atRA), have emerged as key factors in immunity by specifying the localization of immune cells to the gut. Although it is appreciated that isomers of retinoic acid activate the retinoic acid receptor (RAR) and retinoid X receptor (RXR) family of nuclear receptors to elicit cellular changes, the molecular details of retinoic acid action remain poorly defined in immune processes. Here we employ a battery of agonists and antagonists to delineate the specific nuclear receptors utilized by retinoids to evoke lymphocyte cell adhesion to ADAM (adisintegrin and metalloprotease) protein family members. We report that RAR agonism is sufficient to promote immune cell adhesion in both immortal and primary immune cells. Interestingly, adhesion occurs independent of integrin function, and mutant studies demonstrate that atRA-induced adhesion to ADAM members required a distinct binding interface(s) as compared to integrin recognition. Anti-inflammatory corticosteroids as well as 1,25-(OH){sub 2}D{sub 3}, a vitamin D metabolite that prompts immune cell trafficking to the skin, potently inhibited the observed adhesion. Finally, our data establish that induced adhesion was specifically attributable to the RAR-α receptor isotype. The current study provides novel molecular resolution as to which nuclear receptors transduce retinoid exposure into immune cell adhesion.

  14. Activity and biological effects of neem products against arthropods of medical and veterinary importance.

    PubMed

    Mulla, M S; Su, T

    1999-06-01

    Botanical insecticides are relatively safe and degradable, and are readily available sources of biopesticides. The most prominent phytochemical pesticides in recent years are those derived from neem trees, which have been studied extensively in the fields of entomology and phytochemistry, and have uses for medicinal and cosmetic purposes. The neem products have been obtained from several species of neem trees in the family Meliaceae. Six species in this family have been the subject of botanical pesticide research. They are Azadirachta indica A. Juss, Azadirachta excelsa Jack, Azadirachta siamens Valeton, Melia azedarach L., Melia toosendan Sieb. and Zucc., and Melia volkensii Gürke. The Meliaceae, especially A. indica (Indian neem tree), contains at least 35 biologically active principles. Azadirachtin is the predominant insecticidal active ingredient in the seed, leaves, and other parts of the neem tree. Azadirachtin and other compounds in neem products exhibit various modes of action against insects such as antifeedancy, growth regulation, fecundity suppression and sterilization, oviposition repellency or attractancy, changes in biological fitness, and blocking development of vector-borne pathogens. Some of these bioactivity parameters of neem products have been investigated at least in some species of insects of medical and veterinary importance, such as mosquitoes, flies, triatomines, cockroaches, fleas, lice, and others. Here we review, synthesize, and analyze published information on the activity, modes of action, and other biological effects of neem products against arthropods of medical and veterinary importance. The amount of information on the activity, use, and application of neem products for the control of disease vectors and human and animal pests is limited. Additional research is needed to determine the potential usefulness of neem products in vector control programs.

  15. Nuclear Fuels.

    ERIC Educational Resources Information Center

    Nash, J. Thomas

    1983-01-01

    Trends in and factors related to the nuclear industry and nuclear fuel production are discussed. Topics addressed include nuclear reactors, survival of the U.S. uranium industry, production costs, budget cuts by the Department of Energy and U.S. Geological survey for resource studies, mining, and research/development activities. (JN)

  16. Nuclear Rocket Facility Decommissioning Project: Controlled Explosive Demolition of Neutron Activated Shield Wall

    SciTech Connect

    Michael R. Kruzic

    2007-09-16

    Located in Area 25 of the Nevada Test Site (NTS), the Test Cell A (TCA) Facility was used in the early to mid-1960s for the testing of nuclear rocket engines, as part of the Nuclear Rocket Development Program, to further space travel. Nuclear rocket testing resulted in the activation of materials around the reactors and the release of fission products and fuel particles in the immediate area. Identified as Corrective Action Unit 115, the TCA facility was decontaminated and decommissioned (D&D) from December 2004 to July 2005 using the Streamlined Approach for Environmental Restoration (SAFER) process, under the ''Federal Facility Agreement and Consent Order''. The SAFER process allows environmental remediation and facility closure activities (i.e., decommissioning) to occur simultaneously provided technical decisions are made by an experienced decision maker within the site conceptual site model, identified in the Data Quality Objective process. Facility closure involved a seven-step decommissioning strategy. Key lessons learned from the project included: (1) Targeted preliminary investigation activities provided a more solid technical approach, reduced surprises and scope creep, and made the working environment safer for the D&D worker. (2) Early identification of risks and uncertainties provided opportunities for risk management and mitigation planning to address challenges and unanticipated conditions. (3) Team reviews provided an excellent mechanism to consider all aspects of the task, integrated safety into activity performance, increase team unity and ''buy-in'' and promoted innovative and time saving ideas. (4) Development of CED protocols ensured safety and control. (5) The same proven D&D strategy is now being employed on the larger ''sister'' facility, Test Cell C.

  17. Respiratory syncytial virus M2-1 protein induces the activation of nuclear factor kappa B

    SciTech Connect

    Reimers, Kerstin . E-mail: reimers.kerstin@mh-hannover.de; Buchholz, Katja; Werchau, Hermann

    2005-01-20

    Respiratory syncytial virus (RSV) induces the production of a number of cytokines and chemokines by activation of nuclear factor kappa B (NF-{kappa}B). The activation of NF-{kappa}B has been shown to depend on viral replication in the infected cells. In this study, we demonstrate that expression of RSV M2-1 protein, a transcriptional processivity and anti-termination factor, is sufficient to activate NF-{kappa}B in A549 cells. Electromobility shift assays show increased NF-{kappa}B complexes in the nuclei of M2-1-expressing cells. M2-1 protein is found in nuclei of M2-1-expressing cells and in RSV-infected cells. Co-immunoprecipitations of nuclear extracts of M2-1-expressing cells and of RSV-infected cells revealed an association of M2-1 with Rel A protein. Furthermore, the activation of NF-{kappa}B depends on the C-terminus of the RSV M2-1 protein, as shown by NF-{kappa}B-induced gene expression of a reporter gene construct.

  18. Nuclear cathepsin L activity is required for cell cycle progression of colorectal carcinoma cells.

    PubMed

    Tamhane, Tripti; Lllukkumbura, Rukshala; Lu, Shiying; Maelandsmo, Gunhild M; Haugen, Mads H; Brix, Klaudia

    2016-03-01

    Prominent tasks of cysteine cathepsins involve endo-lysosomal proteolysis and turnover of extracellular matrix constituents or plasma membrane proteins for maintenance of intestinal homeostasis. Here we report on enhanced levels and altered subcellular localization of distinct cysteine cathepsins in adenocarcinoma tissue in comparison to adjacent normal colon. Immunofluorescence and immunoblotting investigations revealed the presence of cathepsin L in the nuclear compartment in addition to its expected endo-lysosomal localization in colorectal carcinoma cells. Cathepsin L was represented as the full-length protein in the nuclei of HCT116 cells from which stefin B, a potent cathepsin L inhibitor, was absent. Fluorescence activated cell sorting analyses with synchronized cell cultures revealed deceleration of cell cycle progression of HCT116 cells upon inhibition of cathepsin L activity, while expression of cathepsin L-enhanced green fluorescent protein chimeras accelerated S-phase entry. We conclude that the activity of cathepsin L is high in the nucleus of colorectal carcinoma cells because of lacking stefin B inhibitory activity. Furthermore, we hypothesize that nuclear cathepsin L accelerates cell cycle progression of HCT116 cells thereby supporting the notion that cysteine cathepsins may play significant roles in carcinogenesis due to deregulated trafficking.

  19. A mid-infrared look at the dusty nuclear environments of local active galaxies

    NASA Astrophysics Data System (ADS)

    Alonso Herrero, Almudena

    2016-08-01

    Active galactic nuclei are largely explained in the context of a unified theory, by which a geometrically and optically thick torus of gas and dust obscures the AGN central engine. The torus intercepts a substantial amount of flux from the central engine and and reradiates it in the infrared. There are still many open questions about the nature of the torus material and the role of nuclear (< 100 pc) starbursts in feeding and/or obscuring AGNs. Ground-based mid-infrared imaging and spectroscopy on 8-10m class telescopes allow us to study the dusty environments of nearby active galactic nuclei on physical scales of less than 100pc. In this talk I will present results from a mid-infrared sub-arcsecond resolution imaging and spectroscopy survey of a sample of local AGN. The observations were mostly taken with CanariCam on the 10.4m Gran Telescopio Canarias (GTC) through an ESO/GTC large programme and the CanariCam AGN guaranteed time program. I will discuss results on the torus properties of different types of AGN from the modelling of the unresolved infrared emission with the CLUMPY torus models. I will also show that the molecules responsible for the 11.3micron PAH feature survive in the vicinity of the active nucleus and thus this PAH feature can be used to study the nuclear star formation activity in AGN.

  20. Neutron activation analysis via nuclear decay kinetics using gamma-ray spectroscopy at SFU

    NASA Astrophysics Data System (ADS)

    Domingo, Thomas; Chester, Aaron; Starosta, Krzysztof; Williams, Jonathan

    2016-09-01

    Gamma-ray spectroscopy is a powerful tool used in a variety of fields including nuclear and analytical chemistry, environmental science, and health risk management. At SFU, the Germanium detector for Elemental Analysis and Radiation Studies (GEARS), a low-background shielded high-purity germanium gamma-ray detector, has been used recently in all of the above fields. The current project aims to expand upon the number of applications for which GEARS can be used while enhancing its current functionality. A recent addition to the SFU Nuclear Science laboratory is the Thermo Scientific P 385 neutron generator. This device provides a nominal yield of 3 ×108 neutrons/s providing the capacity for neutron activation analysis, opening a major avenue of research at SFU which was previously unavailable. The isotopes created via neutron activation have a wide range of half-lives. To measure and study isotopes with half-lives above a second, a new analogue data acquisition system has been installed on GEARS allowing accurate measurements of decay kinetics. This new functionality enables identification and quantification of the products of neutron activation. Results from the neutron activation analysis of pure metals will be presented.

  1. Measurements of miniature ionization chamber currents in the JSI TRIGA Mark II reactor demonstrate the importance of the delayed contribution to the photon field in nuclear reactors

    NASA Astrophysics Data System (ADS)

    Radulović, Vladimir; Fourmentel, Damien; Barbot, Loïc; Villard, Jean-François; Kaiba, Tanja; Gašper, Žerovnik; Snoj, Luka

    2015-12-01

    The characterization of experimental locations of a research nuclear reactor implies the determination of neutron and photon flux levels within, with the best achievable accuracy. In nuclear reactors, photon fluxes are commonly calculated by Monte Carlo simulations but rarely measured on-line. In this context, experiments were conducted with a miniature gas ionization chamber (MIC) based on miniature fission chamber mechanical parts, recently developed by the CEA (French Atomic Energy and Alternative Energies Commission) irradiated in the core of the Jožef Stefan Institute TRIGA Mark II reactor in Ljubljana, Slovenia. The aim of the study was to compare the measured MIC currents with calculated currents based on simulations with the MCNP6 code. A discrepancy of around 50% was observed between the measured and the calculated currents; in the latter taking into consideration only the prompt photon field. Further experimental measurements of MIC currents following reactor SCRAMs (reactor shutdown with rapid insertions of control rods) provide evidence that over 30% of the total measured signal is due to the delayed photon field, originating from fission and activation products, which are untreated in the calculations. In the comparison between the measured and calculated values, these findings imply an overall discrepancy of less than 20% of the total signal which is still unexplained.

  2. Nuclear responses to MPF activation and inactivation in Xenopus oocytes and early embryos.

    PubMed

    Lohka, M J

    1998-11-01

    The cell cycle of most organisms is highlighted by characteristic changes in the appearance and activity of the nucleus. Structural changes in the nucleus are particularly evident when a cell begins to divide. At this time, the nuclear envelope is disassembled, the chromatin condenses into metaphase chromosomes, and the chromosomes associate with a newly formed spindle. Upon completion of cell division the nuclear envelope reassembles around the chromosomes as they form telophase nuclei, and subsequently interphase nuclei, in the daughter cells. The cytoplasmic control of nuclear behavior has been the theme of Yoshio Masui's research for much of his career. His pioneering demonstration that the cytoplasm of maturing amphibian oocytes causes the resumption of the meiotic cell cycle when it is injected into an immature oocyte provided unequivocal evidence that a cytoplasmic factor could initiate the transition from interphase to metaphase (M-phase) in intact cells. As described in several reviews in this and the previous issue of Biology of the Cell (see Beckhelling and Ford; Duesbery and Vande Woude; Maller), Masui initially called this activity maturation promoting factor (MPF), but when it was realized that it was a ubiquitous regulator of both mitotic and meiotic cell cycles, MPF came to stand for M-phase promoting factor. Biochemical evidence indicates that MPF activity is composed of a mitotic B-type cyclins and cyclin-dependent kinase 1. The increase in the protein kinase activity of cdk1 initiates the changes in the nucleus associated with oocyte maturation and with the entry into mitosis. This article will attempt to provide a brief summary of the responses of the nucleus to the activation of MPF. In addition, the effect of MPF inactivation on nuclear envelope assembly at the end of mitosis will be discussed. This article is written as a tribute to Yoshio Masui on his retirement from the University of Toronto, and as an expression of gratitude for his

  3. Active Degradation Explains the Distribution of Nuclear Proteins during Cellular Senescence

    PubMed Central

    Giampieri, Enrico; De Cecco, Marco; Remondini, Daniel; Sedivy, John; Castellani, Gastone

    2015-01-01

    The amount of cellular proteins is a crucial parameter that is known to vary between cells as a function of the replicative passages, and can be important during physiological aging. The process of protein degradation is known to be performed by a series of enzymatic reactions, ranging from an initial step of protein ubiquitination to their final fragmentation by the proteasome. In this paper we propose a stochastic dynamical model of nuclear proteins concentration resulting from a balance between a constant production of proteins and their degradation by a cooperative enzymatic reaction. The predictions of this model are compared with experimental data obtained by fluorescence measurements of the amount of nuclear proteins in murine tail fibroblast (MTF) undergoing cellular senescence. Our model provides a three-parameter stationary distribution that is in good agreement with the experimental data even during the transition to the senescent state, where the nuclear protein concentration changes abruptly. The estimation of three parameters (cooperativity, saturation threshold, and maximal velocity of the reaction), and their evolution during replicative passages shows that only the maximal velocity varies significantly. Based on our modeling we speculate the reduction of functionality of the protein degradation mechanism as a possible competitive inhibition of the proteasome. PMID:26115222

  4. Bioturbation effects of Corophium volutator: Importance of density and behavioural activity

    NASA Astrophysics Data System (ADS)

    De Backer, A.; Van Coillie, F.; Montserrat, F.; Provoost, P.; Van Colen, C.; Vincx, M.; Degraer, S.

    2011-01-01

    Bioturbation is one of the major processes influencing ecosystem functioning. Population parameters such as species density, burrow density and species-specific life modes, determine the impact of bioturbation on the ecosystem. A laboratory experiment was developed, using microcosms mimicking a marine intertidal sediment-water interface which allowed for quantification of different population parameters. The vertical redistribution, bioturbation rate and maximum penetration depth of two sizes (41 and 129 μm) of luminophores were measured in five treatments (control, low density of burrows with and without Corophium (1989 ind./m 2), and high density of burrows with and without Corophium (14,921 ind./m 2)) after 1, 7 and 14 days. Results indicate that the behavioural activities of Corophium are of the utmost importance in sediment reworking, since they contributed to a five-fold increase in bioturbation rate compared to the passive transport induced by the static structure of the burrows. Furthermore, density is an important parameter because only high densities play a prominent role in particle transport and hence in organic matter processing, while the role of low Corophium densities is limited in sediment reworking. No evidence for differentiation in sediment size fractions was observed. Finally, bioturbation rates in this study were low compared to other studies, and these results suggest an influence of the tidal rhythmicity in the behavioural activity of Corophium on the bioturbation rate.

  5. Salicylates Inhibit Flavivirus Replication Independently of Blocking Nuclear Factor Kappa B Activation

    PubMed Central

    Liao, Ching-Len; Lin, Yi-Ling; Wu, Bi-Ching; Tsao, Chang-Huei; Wang, Mei-Chuan; Liu, Chiu-I; Huang, Yue-Ling; Chen, Jui-Hui; Wang, Jia-Pey; Chen, Li-Kuang

    2001-01-01

    Flaviviruses comprise a positive-sense RNA genome that replicates exclusively in the cytoplasm of infected cells. Whether flaviviruses require an activated nuclear factor(s) to complete their life cycle and trigger apoptosis in infected cells remains elusive. Flavivirus infections quickly activate nuclear factor kappa B (NF-κB), and salicylates have been shown to inhibit NF-κB activation. In this study, we investigated whether salicylates suppress flavivirus replication and virus-induced apoptosis in cultured cells. In a dose-dependent inhibition, we found salicylates within a range of 1 to 5 mM not only restricted flavivirus replication but also abrogated flavivirus-triggered apoptosis. However, flavivirus replication was not affected by a specific NF-κB peptide inhibitor, SN50, and a proteosome inhibitor, lactacystin. Flaviviruses also replicated and triggered apoptosis in cells stably expressing IκBα-ΔN, a dominant-negative mutant that antagonizes NF-κB activation, as readily as in wild-type BHK-21 cells, suggesting that NF-κB activation is not essential for either flavivirus replication or flavivirus-induced apoptosis. Salicylates still diminished flavivirus replication and blocked apoptosis in the same IκBα-ΔN cells. This inhibition of flaviviruses by salicylates could be partially reversed by a specific p38 mitogen-activated protein (MAP) kinase inhibitor, SB203580. Together, these results show that the mechanism by which salicylates suppress flavivirus infection may involve p38 MAP kinase activity but is independent of blocking the NF-κB pathway. PMID:11483726

  6. Premonolayer oxidation of nanostructured gold: an important factor influencing electrocatalytic activity.

    PubMed

    O'Mullane, Anthony P; Ippolito, Samuel J; Sabri, Ylias M; Bansal, Vipul; Bhargava, Suresh K

    2009-04-09

    The study of the electrodeposition of polycrystalline gold in aqueous solution is important from the viewpoint that in electrocatalysis applications ill-defined micro- and nanostructured surfaces are often employed. In this work, the morphology of gold was controlled by the electrodeposition potential and the introduction of Pb(CH3COO)2 x 3H2O into the plating solution to give either smooth or nanostructured gold crystallites or large dendritic structures which have been characterized by scanning electron microscopy (SEM). The latter structures were achieved through a novel in situ galvanic replacement of lead with AuCl4-(aq) during the course of gold electrodeposition. The electrochemical behavior of electrodeposited gold in the double layer region was studied in acidic and alkaline media and related to electrocatalytic performance for the oxidation of hydrogen peroxide and methanol. It was found that electrodeposited gold is a significantly better electrocatalyst than a polished gold electrode; however, performance is highly dependent on the chosen deposition parameters. The fabrication of a deposit with highly active surface states, comparable to those achieved at severely disrupted metal surfaces through thermal and electrochemical methods, does not result in the most effective electrocatalyst. This is due to significant premonolayer oxidation that occurs in the double layer region of the electrodeposited gold. In particular, in alkaline solution, where gold usually shows the most electrocatalytic activity, these active surface states may be overoxidized and inhibit the electrocatalytic reaction. However, the activity and morphology of an electrodeposited film can be tailored whereby electrodeposited gold that exhibits nanostructure within the crystallites on the surface demonstrated enhanced electrocatalytic activity compared to smaller smooth gold crystallites and larger dendritic structures in potential regions well within the double layer region.

  7. Historical record of nuclear activities from (129)I in corals from the northern hemisphere (Philippines).

    PubMed

    Bautista, Angel T; Matsuzaki, Hiroyuki; Siringan, Fernando P

    2016-11-01

    Iodine-129 is a long-lived fission product that is majorly released in human nuclear activities (HNA) such as nuclear bomb testing, nuclear fuel reprocessing, and nuclear accidents. It is a good environmental tracer and former measurements of (129)I in corals from the southern hemisphere show the increasing trend of (129)I concentrations in the marine environment caused by HNA. Here we show time series of (129)I/(127(stable))I isotopic ratios in two coral cores from the northern hemisphere (Philippines) and how these record (129)I released from HNA in even greater, unprecedented detail. Corals were taken from the Pacific Ocean (Baler) and South China Sea (Parola) sides of the Philippines. We observed nearly identical peaks ((129)I/(127)I ∼ 31.5 × 10(-12)) in both the Baler and Parola records, each attributed to the year 1962 - the year with the highest recorded amount of (129)I release from nuclear bomb testing. This 1962 (129)I bomb signal offers a new time marker that can be used to establish or confirm age models of corals, comparable to or possibly better than the well-known coral (14)C bomb peak. We also observed nuclear fuel reprocessing and Chernobyl accident (129)I signals in years 1977, 1980, and 1986, concurrently in Parola and with 9 to 11-year lags in Baler. This discrepancy in timing suggests that (129)I was transported to the South China Sea and Pacific Ocean sides of the Philippines directly from the atmosphere and through prevailing ocean currents, respectively. Lastly, we observed surprisingly high (129)I/(127)I isotopic ratios (i.e., 22.8 to 38.9 × 10(-12)) in the Parola record after the year 1996, which is in contrast to the decreasing trend observed in the Baler record and in published (129)I releases of different HNA. These results possibly indicate the presence of unknown sources of (129)I in the South China Sea region.

  8. The mid-infrared emission of narrow-line active galactic nuclei: Star formation, nuclear activity, and two populations revealed by WISE

    SciTech Connect

    Rosario, David J.; Burtscher, Leonard; Davies, Richard; Genzel, Reinhard; Lutz, Dieter; Tacconi, Linda J.

    2013-12-01

    We explore the nature of the long-wavelength mid-infrared (MIR) emission of a sample of 13,000 local Type II (narrow-line) active galactic nuclei (AGNs) from the Sloan Digital Sky Survey (SDSS) using 12 μm and 22 μm photometry from the WISE all-sky survey. In combination with FIRST 1.4 GHz photometry, we show that AGNs divide into two relatively distinct populations or 'branches' in the plane of MIR and radio luminosity. Seyfert galaxies lie almost exclusively on an MIR-bright branch (Branch A), while low-ionization nuclear emission line galaxies (LINERs) are split evenly into Branch A and the MIR-faint Branch B. We devise various tests to constrain the processes that define the branches, including a comparison to the properties of pure star-forming inactive galaxies on the MIR-radio plane. We demonstrate that the total MIR emission of objects on Branch A, including most Seyfert galaxies, is governed primarily by host star formation, with ≈15% of the 22 μm luminosity coming from AGN-heated dust. This implies that ongoing dusty star formation is a general property of Seyfert host galaxies. We show that the 12 μm broadband luminosity of AGNs on Branch A is suppressed with respect to star-forming galaxies, possibly due to the destruction of PAHs or deeper 10 μm Si absorption in AGNs. We uncover a correlation between the MIR luminosity and [O III] λ5007 luminosity in AGNs. This suggests a relationship between the star formation rate and nuclear luminosity in the AGN population, but we caution on the importance of selection effects inherent to such AGN-dominated emission-line galaxies in driving such a correlation. We highlight the MIR-radio plane as a useful tool in comparative studies of star formation and nuclear activity in AGNs.

  9. Signal Transducer and Activator of Transcription (STAT)-3 Activates Nuclear Factor (NF)-κB in Chronic Lymphocytic Leukemia Cells

    PubMed Central

    Liu, Zhiming; Hazan-Halevy, Inbal; Harris, David M.; Li, Ping; Ferrajoli, Alessandra; Faderl, Stefan; Keating, Michael J.; Estrov, Zeev

    2014-01-01

    Nuclear factor (NF)-κB plays a major role in the pathogenesis of B-cell neoplasms. A broad array of mostly extracellular stimuli has been reported to activate NF-κB, to various degrees, in chronic lymphocytic leukemia (CLL) cells. Because CLL cells harbor high levels of unphosphorylated (U) signal transducer and activator of transcription (STAT)-3 protein and U-STAT3 was reported to activate NF-κB, we sought to determine whether U-STAT3 activates NF-κB in CLL. Using the electrophoretic mobility shift assay (EMSA) we studied peripheral blood low-density cells from 15 patients with CLL and found that CLL cell nuclear extracts from all the samples bound to an NF-κB DNA probe, suggesting that NF-κB is constitutively activated in CLL. Immunoprecipitation studies showed that STAT3 bound NF-κB p65, and confocal microscopy studies detected U-STAT3/NF-κB complexes in the nuclei of CLL cells, thereby confirming these findings. Furthermore, infection of CLL cells with retroviral STAT3-shRNA attenuated the binding of NF-κB to DNA, as assessed by EMSA, and downregulated mRNA levels of NF-κB-regulated genes, as assessed by quantitative polymerase chain reaction. Taken together, our data suggest that U-STAT3 binds to the NF-κB p50/p65 dimers and that the U-STAT3/NF-κB complexes bind to DNA and activate NF-κB-regulated genes in CLL cells. PMID:21364020

  10. Nuclear Infrared Spectral Energy Distribution of Type II Active Galactic Nuclei

    NASA Astrophysics Data System (ADS)

    Videla, Liza; Lira, Paulina; Andrews, Heather; Alonso-Herrero, Almudena; Alexander, David M.; Ward, Martin

    2013-02-01

    We present near- and mid-IR observations of a sample of Seyfert II galaxies drawn from the 12 μm Galaxy sample. The sample was observed in the J, H, K, L, M and N bands. Galaxy surface brightness profiles are modeled using nuclear, bulge, bar (when necessary), and disk components. To check the reliability of our findings, the procedure was tested using Spitzer observations of M 31. Nuclear spectral energy distributions (SEDs) are determined for 34 objects, and optical spectra are presented for 38, including analysis of their stellar populations using the STARLIGHT spectral synthesis code. Emission line diagnostic diagrams are used to discriminate between genuine active galactic nuclei (AGNs) and H II nuclei. Combining our observations with those found in the literature, we have a total of 40 SEDs. It is found that about 40% of the SEDs are characterized by an upturn in the near-IR, which we have quantified as a NIR slope α < 1 for an SED characterized as λf λvpropλα. The three objects with an H II nucleus and two Seyfert nuclei with strong contamination from a circumnuclear also show an upturn. For genuine AGNs, this component could be explained as emission from the accretion disk, a jet, or from a very hot dust component leaking from the central region through a clumpy obscuring structure. The presence of a very compact nuclear starburst as the origin for this NIR excess emission is not favored by our spectroscopic data for these objects.

  11. Activities ONDRAF/NIRAS related to the decommissioning of nuclear facilities

    SciTech Connect

    Cantarella, J.; Simenon, R.; Braeckeveldt, M.

    2013-07-01

    Since 1980, the Agency is responsible by law for the safe management of all radioactive waste produced in Belgium, including decommissioning wastes. By the law of 11 January 1991 and the implementing Royal Decree of 16/10/1991, ONDRAF/NIRAS has been entrusted with a mission concerning the decommissioning of nuclear facilities. This mission involves the collection and assessment of data concerning decommissioning forecasts for nuclear facilities, the approval of facilities' decommissioning programmes, the establishment - in consultation with operators - of financing conditions for decommissioning, as well as the implementation of these programmes on request by the operator, or in the case of its failure to do so. This is the case for the company Best Medical Belgium SA located at Fleurus (MDS Nordion SA, till April 2011), which produced radioisotopes for medical applications and went bankrupt in 2012. These installations have been entrusted to ONDRAF/NIRAS. A plan of action was developed for taking over the operations in the framework of remediation and decommissioning. Steps have been taken to integrate his new role as a nuclear operator. The installations of Best Medical Belgium SA are now referred to as the 'O/N - Site Fleurus.' Nuclear facility operators, or any person requesting to operate a nuclear facility, are obliged to provide ONDRAF/NIRAS, under their responsibility and in due time, with all the necessary information concerning these facilities' decommissioning forecasts, the nature, quantities and dates of transfer of the resulting waste, and the financing conditions for these facilities' decommissioning. In order to make the necessary funds available for decommissioning a nuclear facility when it will be shut down, operators are obliged to establish provisions during the facility's active life. These provisions are calculated in such a way that the total amount established at the time of the final shutdown covers all costs resulting from the facility

  12. Structural features important for the biological activity of the potassium channel blocking dendrotoxins.

    PubMed

    Hollecker, M; Marshall, D L; Harvey, A L

    1993-10-01

    1. Dendrotoxins from mamba snake venoms are small proteins that block neuronal K+ channels. In order to investigate structural features associated with their biological activity, partially folded versions of dendrotoxins I and K from black mamba (Dendroaspis polylepis) were prepared by selectively reducing one or more of their three S-S bonds. 2. The modified toxins were tested for ability to compete with 125I-labelled native toxin I to high affinity binding sites on rat brain synaptosomal membranes and for the ability to increase acetylcholine release in a neuromuscular preparation. 3. Binding affinity increased progressively as the toxins folded to the native conformation and the most biologically active of the modified species were those in which only the disulphide bond between residues 14 and 38 was not formed. These intermediates had native-like conformations as determined by circular dichroism but still had about 5-10 times lower affinity than native toxins. 4. Addition of negatively charged groups to block the free sulthydryls at positions 14 and 38 caused a further, marked loss of activity. 5. The results are consistent with the existence of two important regions in the dendrotoxin molecules. The region containing two of the disulphide bonds (around Cys5-Cys55 and Cys30-Cys51) and much of the secondary structure is essential for the binding affinity of the toxins, while the region around Cys14 and Cys38, equivalent to part of the antiprotease site of the homologous protease inhibitor from bovine pancreas (BPTI), plays an important role in the potency of dendrotoxins.

  13. Importance of being Nernst: Synaptic activity and functional relevance in stem cell-derived neurons

    PubMed Central

    Bradford, Aaron B; McNutt, Patrick M

    2015-01-01

    Functional synaptogenesis and network emergence are signature endpoints of neurogenesis. These behaviors provide higher-order confirmation that biochemical and cellular processes necessary for neurotransmitter release, post-synaptic detection and network propagation of neuronal activity have been properly expressed and coordinated among cells. The development of synaptic neurotransmission can therefore be considered a defining property of neurons. Although dissociated primary neuron cultures readily form functioning synapses and network behaviors in vitro, continuously cultured neurogenic cell lines have historically failed to meet these criteria. Therefore, in vitro-derived neuron models that develop synaptic transmission are critically needed for a wide array of studies, including molecular neuroscience, developmental neurogenesis, disease research and neurotoxicology. Over the last decade, neurons derived from various stem cell lines have shown varying ability to develop into functionally mature neurons. In this review, we will discuss the neurogenic potential of various stem cells populations, addressing strengths and weaknesses of each, with particular attention to the emergence of functional behaviors. We will propose methods to functionally characterize new stem cell-derived neuron (SCN) platforms to improve their reliability as physiological relevant models. Finally, we will review how synaptically active SCNs can be applied to accelerate research in a variety of areas. Ultimately, emphasizing the critical importance of synaptic activity and network responses as a marker of neuronal maturation is anticipated to result in in vitro findings that better translate to efficacious clinical treatments. PMID:26240679

  14. Activity of bifenthrin, chlorfenapyr, fipronil, and thiamethoxam against red imported fire ants (Hymenoptera: Formicidae).

    PubMed

    Wiltz, B A; Suiter, D R; Gardner, W A

    2010-06-01

    Bifenthrin, chlorfenapyr, fipronil, and thiamethoxam were evaluated for activity against the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae). Mobility impairment and lethal times were determined after topical treatments. Both immobilization and mortality occurred most quickly with bifenthrin, followed by thiamethoxam, chlorfenapyr, and fipronil. Mortality due to horizontal exposure was evaluated at 10, 20, or 30 degrees C, with three ratios of topically treated donor ant corpses to live recipients (5, 10, or 20% donors). Bifenthrin had the greatest horizontal activity of the chemicals tested. For chlorfenapyr, the only treatments having higher mortality than controls were the highest percentage donors at either 10 or 30 degrees C. Horizontal activity of fipronil was temperature dependent only with the highest proportion of donors and was lower than that ofbifenthrin but higher than that of chlorfenapyr or thiamethoxam. Mean mortality due to thiamethoxam was similar to that with chlorfenapyr. Significant mortality occurred in all of the 20 and 30 degrees C thiamethoxam treatments, but none of the 10 degrees C treatments. Effectiveness as a barrier was evaluated by providing a choice between bridges treated with insecticide or water. Although bifenthrin did not provide an impenetrable barrier, it was the only treatment having fewer ants than its paired control bridge. Mortality data suggest that a reduction in recruitment rather than repellency account for this result.

  15. A novel mechanism for regulating the activity of proliferating cell nuclear antigen by a small protein.

    PubMed

    Li, Zhuo; Huang, Richard Y-C; Yopp, Daniel C; Hileman, Travis H; Santangelo, Thomas J; Hurwitz, Jerard; Hudgens, Jeffrey W; Kelman, Zvi

    2014-05-01

    Proliferating cell nuclear antigen (PCNA) forms a trimeric ring that associates with and influences the activity of many proteins participating in DNA metabolic processes and cell cycle progression. Previously, an uncharacterized small protein, encoded by TK0808 in the archaeon Thermococcus kodakarensis, was shown to stably interact with PCNA in vivo. Here, we show that this protein, designated Thermococcales inhibitor of PCNA (TIP), binds to PCNA in vitro and inhibits PCNA-dependent activities likely by preventing PCNA trimerization. Using hydrogen/deuterium exchange mass spectrometry and site-directed mutagenesis, the interacting regions of PCNA and TIP were identified. Most proteins bind to PCNA via a PCNA-interacting peptide (PIP) motif that interacts with the inter domain connecting loop (IDCL) on PCNA. TIP, however, lacks any known PCNA-interacting motif, suggesting a new mechanism for PCNA binding and regulation of PCNA-dependent activities, which may support the development of a new subclass of therapeutic biomolecules for inhibiting PCNA.

  16. The nuclear xenobiotic receptor CAR: structural determinants of constitutive activation and heterodimerization.

    PubMed

    Suino, Kelly; Peng, Li; Reynolds, Ross; Li, Yong; Cha, Ji-Young; Repa, Joyce J; Kliewer, Steven A; Xu, H Eric

    2004-12-22

    Constitutive androstane receptor (CAR) induces xenobiotic, bilirubin, and thyroid hormone metabolism as a heterodimer with the retinoid X receptor (RXR). Unlike ligand-dependent nuclear receptors, CAR is constitutively active. Here, we report the heterodimeric structure of the CAR and RXR ligand binding domains (LBDs), which reveals an unusually large dimerization interface and a small CAR ligand binding pocket. Constitutive CAR activity appears to be mediated by the compact nature of the CAR LBD that displays several unique features including a shortened AF2 helix and helix H10, which are linked by a two-turn helix that normally adopts an extended loop in other receptors, and an extended helix H2 that stabilizes the canonical LBD fold by packing tightly against helix H3. These structural observations provide a molecular framework for understanding the atypical transcriptional activation properties of CAR.

  17. Ligand-binding domains of nuclear receptors facilitate tight control of split CRISPR activity

    PubMed Central

    Nguyen, Duy P.; Miyaoka, Yuichiro; Gilbert, Luke A.; Mayerl, Steven J.; Lee, Brian H.; Weissman, Jonathan S.; Conklin, Bruce R.; Wells, James A.

    2016-01-01

    Cas9-based RNA-guided nuclease (RGN) has emerged to be a versatile method for genome editing due to the ease of construction of RGN reagents to target specific genomic sequences. The ability to control the activity of Cas9 with a high temporal resolution will facilitate tight regulation of genome editing processes for studying the dynamics of transcriptional regulation or epigenetic modifications in complex biological systems. Here we show that fusing ligand-binding domains of nuclear receptors to split Cas9 protein fragments can provide chemical control over split Cas9 activity. The method has allowed us to control Cas9 activity in a tunable manner with no significant background, which has been challenging for other inducible Cas9 constructs. We anticipate that our design will provide opportunities through the use of different ligand-binding domains to enable multiplexed genome regulation of endogenous genes in distinct loci through simultaneous chemical regulation of orthogonal Cas9 variants. PMID:27363581

  18. Discovery of a Novel, Isothiazolonaphthoquinone-Based Small Molecule Activator of FOXO Nuclear-Cytoplasmic Shuttling

    PubMed Central

    Cautain, Bastien; Castillo, Francisco; Musso, Loana; Ferreira, Bibiana I.; de Pedro, Nuria; Rodriguez Quesada, Lorena; Machado, Susana; Vicente, Francisca; Dallavalle, Sabrina

    2016-01-01

    FOXO factors are tumour suppressor proteins commonly inactivated in human tumours by posttranslational modifications. Furthermore, genetic variation within the FOXO3a gene is consistently associated with human longevity. Therefore, the pharmacological activation of FOXO proteins is considered as an attractive therapeutic approach to treat cancer and age-related diseases. In order to identify agents capable of activating FOXOs, we tested a collection of small chemical compounds using image-based high content screening technology. Here, we report the discovery of LOM612 (compound 1a), a newly synthesized isothiazolonaphthoquinone as a potent FOXO relocator. Compound 1a induces nuclear translocation of a FOXO3a reporter protein as well as endogenous FOXO3a and FOXO1 in U2OS cells in a dose-dependent manner. This activity does not affect the subcellular localization of other cellular proteins including NFkB or inhibit CRM1-mediated nuclear export. Furthermore, compound 1a shows a potent antiproliferative effect in human cancer cell lines. PMID:27936162

  19. Instrumental nuclear activation analysis (INAA) characterization of environmental air filter samples.

    PubMed

    Alemón, Ernesto; Herrera, Luis; Ortiz, Elba; Longoria, L C Luis C

    2004-06-01

    Nuclear techniques have been used in quantitations of environmental pollutants, and instrumental neutron activation analysis (INAA) has turned out to be particularly useful in the analysis of airborne suspended particles. This work describes the INAA characterization of the particulate material in the environmental samples obtained in a monitoring campaign in Mexico City's Metropolitan Area. As the types of the irradiation facilities and gamma-ray detection system impose some limitations on the possibilities of INAA analysis, the actual experimental conditions at Gamma Spectroscopy Laboratory, where the analysis was performed, had been assessed. The facilities had been found suitable for the analysis of samples from this campaign, in which 22 elements were determined.

  20. Results from the Nuclear Plant Aging Research Program: Their use in inspection activities

    SciTech Connect

    Gunther, W.; Taylor, J. )

    1990-09-01

    The US NCR's Nuclear Plant Aging Research (NPAR) Program has determined the susceptibility to aging of components and systems, and the potential for aging to impact plant safety and availability. The NPAR Program also identified methods for detecting and mitigating aging in components. This report describes the NPAR results which can enhance NRC inspection activities. Recommendations are provided for communicating pertinent information to NRC inspectors. These recommendations are based on a detailed assessment of the NRC's Inspection Program, and feedback from resident and regional inspectors as described within. Examples of NPAR report summaries and aging inspection guides for components and systems are included. 13 refs., 3 figs., 4 tabs.

  1. History of the bubble chamber and related active- and internal-target nuclear tracking detectors

    NASA Astrophysics Data System (ADS)

    Becchetti, F. D.

    2015-06-01

    Donald Glaser, 1960 Nobel laureate in Physics, recently passed away (2013), as have many of his colleagues who were involved with the early development of bubble chambers at the University of Michigan. In this paper I will review those early years and the subsequent wide-spread application of active-target (AT) bubble chambers that dominated high-energy physics (HEP) research for over thirty years. Some of the related, but more modern nuclear tracking detectors being used in HEP, neutrino astrophysics and dark-matter searches also will be discussed.

  2. Thoron activity level and radon measurement by a nuclear track detector.

    PubMed

    Planinić, J; Faj, Z; Vuković, B

    1993-03-01

    Radon activity concentrations in the air were measured with LR-115 nuclear track detectors at three locations in Osijek. The respective equilibrium factors and the effective dose equivalents were determined. Indoor concentrations were from 9.8 to 58.2 Bq m-3 and relative errors of the track etching method were near 19 per cent. The indoor alpha potential energy of the radon and thoron progenies was measured with an ISD detector. Independent measurements, performed with a Radhome semiconductor detector, showed that the indoor thoron concentration was nearly 20 per cent of the radon one.

  3. Transcriptional activation of NAD{sup +}-dependent protein deacetylase SIRT1 by nuclear receptor TLX

    SciTech Connect

    Iwahara, Naotoshi; Hisahara, Shin; Hayashi, Takashi; Horio, Yoshiyuki

    2009-09-04

    An orphan nuclear receptor TLX is a transcriptional repressor that promotes the proliferation and self-renewal of neural precursor cells (NPCs). SIRT1, an NAD{sup +}-dependent protein deacetylase, is highly expressed in the NPCs and participates in neurogenesis. Here, we found that TLX colocalized with SIRT1 and knockdown of TLX by small interfering RNAs decreased SIRT1 levels in NPCs. TLX increased the SIRT1 expression by binding to the newly identified TLX-activating element in the SIRT1 gene promoter in HEK293 cells. Thus, TLX is an inducer of SIRT1 and may contribute to neurogenesis both as a transactivator and as a repressor.

  4. Differentiation between earthquake radon anomalies and those arising from nuclear activities.

    PubMed

    Elmaghraby, Elsayed K; Lotfy, Yahia A

    2009-01-01

    The present work offers a methodology to correlate the earthquake time, magnitude and location based on the behavior of radon concentration. By correlating tectonic events to radon behavior, based on previously published radon data, radon concentration signals can be taken into consideration in decision making. Three correlations between magnitude, time and epicenter distance are presented. The applicability of any one of them depends on the measurement technique. Moreover, a suggested procedure is presented to determine the possible confusion between signals for radon anomalies originating from tectonic failure and those arising from nuclear activities. The suggested method depends on the change observed on the (219)Rn and (222)Rn daughters concentration ratios.

  5. AGN POPULATION IN HICKSON COMPACT GROUPS. I. DATA AND NUCLEAR ACTIVITY CLASSIFICATION

    SciTech Connect

    MartInez, M. A.; Del Olmo, A.; Perea, J.; Coziol, R. E-mail: chony@iaa.es E-mail: rcoziol@astro.ugto.mx

    2010-03-15

    We have conducted a new spectroscopic survey to characterize the nature of nuclear activity in Hickson compact group (HCG) galaxies and establish its frequency. We have obtained new intermediate-resolution optical spectroscopy for 200 member galaxies and corrected for underlying stellar population contamination using galaxy templates. Spectra for 11 additional galaxies have been acquired from the ESO and 6dF public archives, and emission-line ratios have been taken from the literature for 59 more galaxies. Here we present the results of our classification of the nuclear activity for 270 member galaxies, which belong to a well-defined sample of 64 HCGs. We found a large fraction of galaxies, 63%, with emission lines. Using standard diagnostic diagrams, 45% of the emission-line galaxies were classified as pure active galactic nuclei (AGNs), 23% as Transition Objects (TOs), and 32% as star-forming nuclei (SFNs). In the HCGs, the AGN activity appears as the most frequent activity type. Adopting the interpretation that in TOs a low-luminosity AGN coexists with circumnuclear star formation, the fraction of galaxies with an AGN could rise to 42% of the whole sample. The low frequency (20%) of SFNs confirms that there is no star formation enhancement in HCGs. After extinction correction, we found a median AGN H{alpha} luminosity of 7.1 x 10{sup 39} erg s{sup -1}, which implies that AGNs in HCG have a characteristically low luminosity. This result added to the fact that there is an almost complete absence of broad-line AGNs in compact groups (CGs) as found by MartInez et al. and corroborated in this study for HCGs, is consistent with very few gas left in these galaxies. In general, therefore, what may characterize the level of activity in CGs is a severe deficiency of gas.

  6. Molecular mechanisms of action of the soy isoflavones includes activation of promiscuous nuclear receptors. A review.

    PubMed

    Ricketts, Marie-Louise; Moore, David D; Banz, William J; Mezei, Orsolya; Shay, Neil F

    2005-06-01

    Consumption of soy has been demonstrated to reduce circulating cholesterol levels, most notably reducing low-density lipoprotein (LDL) cholesterol levels in hypercholesterolemic individuals. The component or components that might be responsible for this effect is still a matter of debate or controversy among many researchers. Candidate agents include an activity of soy protein itself, bioactive peptides produced during the digestive process, or the soy isoflavones. Although soy intake may provide other health benefits including preventative or remediative effects on cancer, osteoporosis and symptoms of menopause, this review will focus on isoflavones as agents affecting lipid metabolism. Isoflavones were first discovered as a bioactive agent disrupting estrogen action in female sheep, thereby earning the often-used term 'phytoestrogens'. Subsequent work confirmed the ability of isoflavones to bind to estrogen receptors. Along with the cholesterol-lowering effect of soy intake, research that is more recent has pointed to a beneficial antidiabetic effect of soy intake, perhaps mediated by soy isoflavones. The two common categories of antidiabetic drugs acting on nuclear receptors known as peroxisome proliferator activated receptors (PPARs) are the fibrates and glitazones. We and others have recently asked the research question 'do the soy isoflavones have activities as either "phytofibrates" or "phytoglitazones"?' Such an activity should be able to be confirmed both in vivo and in vitro. In both the in vivo and in vitro cases, this action has indeed been confirmed. Further work suggests a possible action of isoflavones similar to the nonestrogenic ligands that bind the estrogen-related receptors (ERRs). Recently, these receptors have been demonstrated to contribute to lipolytic processes. Finally, evaluation of receptor activation studies suggests that thyroid receptor activation may provide additional clues explaining the metabolic action of isoflavones. The recent

  7. Glycogen synthase kinase 3{beta} regulation of nuclear factor of activated T-cells isoform c1 in the vascular smooth muscle cell response to injury

    SciTech Connect

    Chow Winsion; Hou Guangpei; Bendeck, Michelle P.

    2008-10-01

    The migration and proliferation of vascular smooth muscle cells (vSMCs) are critical events in neointima formation during atherosclerosis and restenosis. The transcription factor nuclear factor of activated T-cells-isoform c1 (NFATc1) is regulated by atherogenic cytokines, and has been implicated in the migratory and proliferative responses of vSMCs through the regulation of gene expression. In T-cells, calcineurin de-phosphorylates NFATc1, leading to its nuclear import, while glycogen synthase kinase 3 {beta} (GSK3{beta}) phosphorylates NFATc1 and promotes its nuclear export. However, the relationship between NFATc1 and GSK3{beta} has not been studied during SMC migration and proliferation. We investigated this by scrape wounding vSMCs in vitro, and studying wound repair. NFATc1 protein was transiently increased, reaching a peak at 8 h after wounding. Cell fractionation and immunocytochemistry revealed that NFATc1 accumulation in the nucleus was maximal at 4 h after injury, and this was coincident with a significant 9 fold increase in transcriptional activity. Silencing NFATc1 expression with siRNA or inhibition of NFAT with cyclosporin A (CsA) attenuated wound closure by vSMCs. Phospho-GSK3{beta} (inactive) increased to a peak at 30 min after injury, preceding the nuclear accumulation of NFATc1. Overexpression of a constitutively active mutant of GSK3{beta} delayed the nuclear accumulation of NFATc1, caused a 50% decrease in NFAT transcriptional activity, and attenuated vSMC wound repair. We conclude that NFATc1 promotes the vSMC response to injury, and that inhibition of GSK3{beta} is required for the activation of NFAT during wound repair.

  8. Role of Calcineurin, hnRNPA2 and Akt in Mitochondrial Respiratory Stress-Mediated Transcription Activation of Nuclear Gene Targets

    PubMed Central

    Guha, Manti; Tang, Weigang; Sondheimer, Neal; Avadhani, Narayan G.

    2010-01-01

    Pathophysiological conditions causing mitochondrial dysfunction and altered transmembrane potential (Δψm) initiate a mitochondrial respiratory stress response, also known as mitochondrial retrograde response, in a variety of mammalian cells. An increase in the cytosolic Ca2+ [Ca2+]c as part of this signaling cascade activates Ca2+ responsive phosphatase, Calcineurin (Cn). Activation of IGF1R accompanied by increased glycolysis, invasiveness, and resistance to apoptosis are phenotypic hallmarks of C2C12 rhabdomyoblast cells subjected to this stress. The signaling is associated with activation and increased nuclear translocation of a number of transcription factors including a novel NFκB (cRel: p50) pathway, NFAT, CREB and C/EBPδ. This culminates in the upregulation of a number of nuclear genes including Cathepsin L, RyR1, Glut4 and Akt1. We observed that stress regulated transcription activation of nuclear genes involves a cooperative interplay between NFκB (cRel:p50), C/EBPδ, CREB, NFAT. Our results show that the functional synergy of these factors requires the stress-activated heterogeneous nuclear ribonucleoprotein, hnRNPA2 as a transcriptional co-activator. We report here that mitochondrial stress leads to induced expression and activation of serine threonine kinase Akt1. Interestingly, we observe that Akt1 phosphorylates hnRNPA2 under mitochondrial stress conditions, which is a crucial step for the recruitment of this coactivator to the stress target promoters and culmination in mitochondrial stress-mediated transcription activation of target genes. We propose that mitochondrial stress plays an important role in tumor progression and emergence of invasive phenotypes. PMID:20153290

  9. Advanced x-ray spectrometric techniques for characterization of nuclear materials: An overview of recent laboratory activities

    NASA Astrophysics Data System (ADS)

    Misra, N. L.

    2014-11-01

    Advancements in x-ray spectrometric techniques at different stages have made this technique suitable for characterization of nuclear materials with respect to trace/major element determinations and compositional uniformity studies. The two important features of total reflection x-ray fluorescence spectrometry: 1) requirement of very small amount of sample in ng level 2) multielement analytical capability, in addition to other features, make this technique very much suitable to nuclear materials characterization as most of the nuclear materials are radioactive and the radioactive waste generated and radiation hazards to the operator are minimum when such low amount of sample is used. Similarly advanced features of energy dispersive x-ray fluorescence e.g. better geometry for high flux, reduction in background due to application of radiation filters have made the measurements of samples sealed inside thin alkathene/PVC covers possible with good sensitivity. This approach avoids putting the instrument inside a glove box for measuring radioactive samples and makes the operation/maintenance of the instrument and analysis of the samples possible in easy and fast manner. This approach has been used for major element determinations in mixed uranium-plutonium samples. Similarly μ-XRF with brilliant and micro-focused excitation sources can be used for compositional uniformity study of reactor fuel pellets. A μ-XRF study using synchrotron light source has been made to assess the compositional uniformity of mixed uranium-thorium oxide pellets produced by different processes. This approach is simple as it does not involve any sample preparation and is non-destructive. A brief summary of such activities carried out in our laboratory in past as well as ongoing and planned for the future have been discussed in the present manuscript.

  10. Increasing ocean temperatures reduce activity patterns of a large commercially important coral reef fish.

    PubMed

    Johansen, J L; Messmer, V; Coker, D J; Hoey, A S; Pratchett, M S

    2014-04-01

    Large-bodied fish are critical for sustaining coral reef fisheries, but little is known about the vulnerability of these fish to global warming. This study examined the effects of elevated temperatures on the movement and activity patterns of the common coral trout Plectropomus leopardus (Serranidae), which is an important fishery species in tropical Australia and throughout the Indo West-Pacific. Adult fish were collected from two locations on Australia's Great Barrier Reef (23°S and 14°S) and maintained at one of four temperatures (24, 27, 30, 33 °C). Following >4 weeks acclimation, the spontaneous swimming speeds and activity patterns of individuals were recorded over a period of 12 days. At 24-27 °C, spontaneous swimming speeds of common coral trout were 0.43-0.45 body lengths per second (bls(-1)), but dropped sharply to 0.29 bls(-1) at 30 °C and 0.25 bls(-1) at 33 °C. Concurrently, individuals spent 9.3-10.6% of their time resting motionless on the bottom at 24-27 °C, but this behaviour increased to 14.0% at 30 °C and 20.0% of the time at 33 °C (mean ± SE). The impact of temperature was greatest for smaller individuals (<45 cm TL), showing significant changes to swimming speeds across every temperature tested, while medium (45-55 cm TL) and large individuals (>55 cm TL) were first affected by 30 °C and 33 °C, respectively. Importantly, there was some indication that populations can adapt to elevated temperature if presented with adequate time, as the high-latitude population decreased significantly in swimming speeds at both 30 °C and 33 °C, while the low-latitude population only showed significant reductions at 33 °C. Given that movement and activity patterns of large mobile species are directly related to prey encounter rates, ability to capture prey and avoid predators, any reductions in activity patterns are likely to reduce overall foraging and energy intake, limit the energy available for growth and reproduction, and affect the fitness and

  11. Nuclear transfer in rodents.

    PubMed

    Mullins, Linda J; Wilmut, Ian; Mullins, John J

    2004-01-01

    Cloning is the asexual reproduction of an individual, such that the offspring have an essentially identical nuclear genome. Nuclear transfer and cloning have been achieved in a number of species, namely sheep, cows, goats, rabbits, cats and mice, but have been largely unsuccessful, so far, in dogs, primates and rats. Clearly, contributory factors which affect the outcome of successful cloning experiments are not universally applicable to all species. One theme common to all cloning experiments, however, is the overall inefficiency of the process, typically 0-4%. A number of factors contribute to nuclear transfer inefficiency, and we will review mouse cloning experiments, which address these problems, highlighting the importance of donor nucleus choice (somatic or ES cell, fetal or adult, quiescent or actively dividing). Finally, we will summarize the emerging principles which appear to govern nuclear reprogramming and production of clones, and will consider the application of nuclear transfer to the rat.

  12. The role of constitutive nitric-oxide synthase in ultraviolet B light-induced nuclear factor κB activity.

    PubMed

    Tong, Lingying; Wu, Shiyong

    2014-09-19

    NF-κB is a transcription factor involved in many signaling pathways that also plays an important role in UV-induced skin tumorigenesis. UV radiation can activate NF-κB, but the detailed mechanism remains unclear. In this study, we provided evidence that the activation of constitutive nitric-oxide synthase plays a role in regulation of IκB reduction and NF-κB activation in human keratinocyte HaCaT cells in early phase (within 6 h) post-UVB. Treating the cells with l-NAME, a selective inhibitor of constitutive nitric-oxide synthase (cNOS), can partially reverse the IκB reduction and inhibit the DNA binding activity as well as nuclear translocation of NF-κB after UVB radiation. A luciferase reporter assay indicates that UVB-induced NF-κB activation is totally diminished in cNOS null cells. The cNOS-mediated reduction of IκB is likely due to the imbalance of nitric oxide/peroxynitrite because treating the cells with lower (50 μm), but not higher (100-500 μm), concentration of S-nitroso-N-acetylpenicillamine (SNAP) can reverse the effect of l-NAME in partial restore IκB level post-UVB. Our data also showed that NF-κB activity was required for maintaining a stable IκB kinase α subunit (IKKα) level because treating the cells with NF-κB or cNOS inhibitors could reduce IKKα level upon UVB radiation. In addition, our data demonstrated that although NF-κB protects cells from UVB-induced death, its pro-survival activity was likely neutralized by the pro-death activity of peroxynitrite after UVB radiation.

  13. Importance of neutron energy distribution in borehole activation analysis in relatively dry, low-porosity rocks

    USGS Publications Warehouse

    Senftle, F.E.; Moxham, R.M.; Tanner, A.B.; Philbin, P.W.; Boynton, G.R.; Wager, R.E.

    1977-01-01

    To evaluate the importance of variations in the neutron energy distribution in borehole activation analysis, capture gamma-ray measurements were made in relatively dry, low-porosity gabbro of the Duluth Complex. Although sections of over a meter of solid rock were encountered in the borehole, there was significant fracturing with interstitial water leading to a substantial variation of water with depth in the borehole. The linear-correlation coefficients calculated for the peak intensities of several elements compared to the chemical core analyses were generally poor throughout the depth investigated. The data suggest and arguments are given which indicate that the variation of the thermal-to-intermediate-to-fast neutron flux density as a function of borehole depth is a serious source of error and is a major cause of the changes observed in the capture gamma-ray peak intensities. These variations in neutron energy may also cause a shift in the observed capture gamma-ray energy.

  14. [Antibacterial and antifungal activity of Salvia apiana against clinically important microorganisms].

    PubMed

    Córdova-Guerrero, Iván; Aragon-Martinez, Othoniel H; Díaz-Rubio, Laura; Franco-Cabrera, Santiago; Serafín-Higuera, Nicolas A; Pozos-Guillén, Amaury; Soto-Castro, Tely A; Martinez-Morales, Flavio; Isiordia-Espinoza, Mario

    Due to the great global concern regarding bacterial resistance to antibiotics, an ongoing search for new molecules having antibacterial activity is necessary. This study evaluated the antibacterial and anticandidal effects of a hexane extract from the root of Salvia apiana. Salvia extracts at concentrations of 27, 13.5, 6.8 and 3.4mg/ml caused growth inhibition of Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis and Candida albicans. However, no significant effect was observed on Escherichia coli and Candida tropicalis in comparison to vehicle. It was here demonstrated for the first time that Salvia apiana has an important antimicrobial effect on human pathogens of great clinical value, thus opening the field to continue the evaluation of this lamiaceous plant for its future use as a therapeutic agent.

  15. Expanded polyglutamine domain possesses nuclear export activity which modulates subcellular localization and toxicity of polyQ disease protein via exportin-1.

    PubMed

    Chan, Wing Man; Tsoi, Ho; Wu, Chi Chung; Wong, Chi Hang; Cheng, Tat Cheung; Li, Hoi Yeung; Lau, Kwok Fai; Shaw, Pang Chui; Perrimon, Norbert; Chan, Ho Yin Edwin

    2011-05-01

    Polyglutamine (polyQ) diseases are a group of late-onset, progressive neurodegenerative disorders caused by CAG trinucleotide repeat expansion in the coding region of disease genes. The cell nucleus is an important site of pathology in polyQ diseases, and transcriptional dysregulation is one of the pathologic hallmarks observed. In this study, we showed that exportin-1 (Xpo1) regulates the nucleocytoplasmic distribution of expanded polyQ protein. We found that expanded polyQ protein, but not its unexpanded form, possesses nuclear export activity and interacts with Xpo1. Genetic manipulation of Xpo1 expression levels in transgenic Drosophila models of polyQ disease confirmed the specific nuclear export role of Xpo1 on expanded polyQ protein. Upon Xpo1 knockdown, the expanded polyQ protein was retained in the nucleus. The nuclear disease protein enhanced polyQ toxicity by binding to heat shock protein (hsp) gene promoter and abolished hsp gene induction. Further, we uncovered a developmental decline of Xpo1 protein levels in vivo that contributes to the accumulation of expanded polyQ protein in the nucleus of symptomatic polyQ transgenic mice. Taken together, we first showed that Xpo1 is a nuclear export receptor for expanded polyQ domain, and our findings establish a direct link between protein nuclear export and the progressive nature of polyQ neurodegeneration.

  16. Blockade of T-cell activation by dithiocarbamates involves novel mechanisms of inhibition of nuclear factor of activated T cells.

    PubMed Central

    Martínez-Martínez, S; Gómez del Arco, P; Armesilla, A L; Aramburu, J; Luo, C; Rao, A; Redondo, J M

    1997-01-01

    Dithiocarbamates (DTCs) have recently been reported as powerful inhibitors of NF-kappaB activation in a number of cell types. Given the role of this transcription factor in the regulation of gene expression in the inflammatory response, NF-kappaB inhibitors have been suggested as potential therapeutic drugs for inflammatory diseases. We show here that DTCs inhibited both interleukin 2 (IL-2) synthesis and membrane expression of antigens which are induced during T-cell activation. This inhibition, which occurred with a parallel activation of c-Jun transactivating functions and expression, was reflected by transfection experiments at the IL-2 promoter level, and involved not only the inhibition of NF-kappaB-driven reporter activation but also that of nuclear factor of activated T cells (NFAT). Accordingly, electrophoretic mobility shift assays (EMSAs) indicated that pyrrolidine DTC (PDTC) prevented NF-kappaB, and NFAT DNA-binding activity in T cells stimulated with either phorbol myristate acetate plus ionophore or antibodies against the CD3-T-cell receptor complex and simultaneously activated the binding of AP-1. Furthermore, PDTC differentially targeted both NFATp and NFATc family members, inhibiting the transactivation functions of NFATp and mRNA induction of NFATc. Strikingly, Western blotting and immunocytochemical experiments indicated that PDTC promoted a transient and rapid shuttling of NFATp and NFATc, leading to their accelerated export from the nucleus of activated T cells. We propose that the activation of an NFAT kinase by PDTC could be responsible for the rapid shuttling of the NFAT, therefore transiently converting the sustained transactivation of this transcription factor that occurs during lymphocyte activation, and show that c-Jun NH2-terminal kinase (JNK) can act by directly phosphorylating NFATp. In addition, the combined inhibitory effects on NFAT and NF-KB support a potential use of DTCs as immunosuppressants. PMID:9343406

  17. Hydrothermal activity at slow-spreading ridges: variability and importance of magmatic controls

    NASA Astrophysics Data System (ADS)

    Escartin, Javier

    2016-04-01

    less studied, similar hydrothermal systems are found elsewhere associated to other central volcanoes along the ridge axis (e.g., Menez Gwenn at the Mid-Atlantic Ridge and Soria Mornia or Troll Wall at the Arctic Ridges). Long-lived hydrothermal activity plays an important role in controlling the thermal structure of the lithosphere and its accretion at and near-axis, and also determining the distribution and biogeography of vent communities. Along slow-spreading segments, long-lived hydrothermal activity can be provided both by volcanic systems (e.g., Lucky Strike) and tectonic systems (oceanic detachment faults). While magmatic and hydrothermal activity is relatively well understood now in volcanic systems (e.g., Lucky Strike), tectonic systems (oceanic detachment faults) require further integrated studies to constrain the links between long-lived localization of deformation along oceanic detachment faults, hydrothermal activity, and origin and nature of off-axis heat sources animating hydrothermal circulation.

  18. β-Amyloid induces nuclear protease-mediated lamin fragmentation independent of caspase activation.

    PubMed

    Ramasamy, Vijay Sankar; Islam, Md Imamul; Haque, Md Aminul; Shin, Song Yub; Park, Il-Seon

    2016-06-01

    β-Amyloid (Aβ), a hallmark peptide of Alzheimer's disease, induces both caspase-dependent apoptosis and non-apoptotic cell death. In this study, we examined caspase-independent non-apoptotic cell death preceding caspase activation in Aβ42-treated cells. We first determined the optimal treatment conditions for inducing cell death without caspase activation and selected a double-treatment method involving the incubation of cells with Aβ42 for 4 and 6 h (4+6 h sample). We observed that levels of lamin A (LA) and lamin B (LB) were reduced in the 4+6 h samples. This reduction was decreased by treatment with suc-AAPF-CMK, an inhibitor of nuclear scaffold (NS) protease, but not by treatment with z-VAD-FMK, a pan-caspase inhibitor. In addition, suc-AAPF-CMK decreased the changes in nuclear morphology observed in cells in the 4+6 h samples, which were different from nuclear fragmentation observed in STS-treated cells. Furthermore, suc-AAPF-CMK inhibited cell death in the 4+6 h samples. LA and LB fragmentation occurred in the isolated nuclei and was also inhibited by suc-AAPF-CMK. Together, these data indicated that the fragmentation of LA and LB in the Aβ42-treated cells was induced by an NS protease, whose identity is not clearly determined yet. A correlation between Aβ42 toxicity and the lamin fragmentation by NS protease suggests that inhibition of the protease could be an effective method for controlling the pathological process of AD.

  19. Nuclear activity versus star formation: emission-line diagnostics at ultraviolet and optical wavelengths

    NASA Astrophysics Data System (ADS)

    Feltre, A.; Charlot, S.; Gutkin, J.

    2016-03-01

    In the context of observations of the rest-frame ultraviolet and optical emission from distant galaxies, we explore the emission-line properties of photoionization models of active and inactive galaxies. Our aim is to identify new line-ratio diagnostics to discriminate between gas photoionization by active galactic nuclei (AGN) and star formation. We use a standard photoionization code to compute the emission from AGN narrow-line regions and compare this with calculations of the nebular emission from star-forming galaxies achieved using the same code. We confirm the appropriateness of widely used optical spectral diagnostics of nuclear activity versus star formation and explore new diagnostics at ultraviolet wavelengths. We find that combinations of a collisionally excited metal line or line multiplet, such as C IV λλ1548, 1551, O III] λλ1661, 1666, N III] λ1750, [Si III] λ1883+Si III] λ1892 and [C III] λ1907+C III] λ1909, with the He II λ1640 recombination line are individually good discriminants of the nature of the ionizing source. Diagrams involving at least three of these lines allow an even more stringent distinction between active and inactive galaxies, as well as valuable constraints on interstellar gas parameters and the shape of the ionizing radiation. Several line ratios involving Ne-based emission lines, such as [Ne IV] λ2424, [Ne III] λ3343 and [Ne V] λ3426, are also good diagnostics of nuclear activity. Our results provide a comprehensive framework to identify the sources of photoionization and physical conditions of the ionized gas from the ultraviolet and optical nebular emission from galaxies. This will be particularly useful to interpret observations of high-redshift galaxies with future facilities, such as the James Webb Space Telescope and extremely large ground-based telescopes.

  20. The importance of self-determined motivation towards physical activity in patients with schizophrenia.

    PubMed

    Vancampfort, Davy; De Hert, Marc; Vansteenkiste, Maarten; De Herdt, Amber; Scheewe, Thomas W; Soundy, Andrew; Stubbs, Brendon; Probst, Michel

    2013-12-30

    There is a need for theoretically-based research on the motivational processes linked to the commencement and continuation of physical activity in patients with schizophrenia. Within the Self-Determination Theory (SDT) framework, we investigated the SDT tenets in these patients by examining the factor structure of the Behavioral Regulation in Exercise Questionnaire-2 (BREQ-2) and by investigating associations between motivation and PA. The secondary aim was to study differences in motivation according to gender, educational level, treatment setting and disease stage. A total of 129 patients (44♀) with schizophrenia agreed to participate. Exploratory factor analysis showed sufficient convergence with the original factor for amotivation, external and introjected regulation, while identified and intrinsic regulations loaded on a single factor which we labeled "autonomous regulation". Significant positive correlations were found between the total physical activity score and the subscales amotivation (r = -0.44, P < 0.001), external regulation (r = -0.27, P < 0.001), and autonomous regulation (r = 0.57, P < 0.001). Outpatients reported more external (P < 0.05) and introjected (P < 0.05) regulations than inpatients. Our results suggest that patients' level of self-determination may play an important role in the adoption and maintenance of health promoting behaviors in patients with schizophrenia.

  1. Identification of a protein with antioxidant activity that is important for the protection against beer ageing.

    PubMed

    Wu, Ming J; Clarke, Frank M; Rogers, Peter J; Young, Paul; Sales, Narelle; O'Doherty, Patrick J; Higgins, Vincent J

    2011-01-01

    This study was carried out with fresh Australian lager beer which was sampled directly off the production line, the same samples aged for 12 weeks at 30 °C, and the vintage beer which was kept at 20 °C for 5 years. Characteristic Australian lager flavour was maintained in the fresh and vintage beers but was lost in the aged beer. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and free thiol group labelling analyses of beer proteins found that this flavour stability correlated with the presence of an unknown 10 kilodaltons (kDa) protein with a higher level of free thiols. The protein was purified by size-exclusion chromatography, then peptide sequencing and database matching identified it as the barley lipid transfer protein (LTP1). Further characterisation using diphenylpicrylhydrazyl (DPPH) free radical scavenging and a Saccharomyces cerevisiae-based antioxidant screening assay demonstrated that the LTP1 protein was active in DPPH reduction and antioxidant activity. The absence of free thiol in the aged beer indicates that the thiol functional groups within the LTP1 protein were saturated and suggests that it is important in the flavour stability of beer by maintaining reduction capacity during the ageing process.

  2. Identification of a Protein with Antioxidant Activity that is Important for the Protection against Beer Ageing

    PubMed Central

    Wu, Ming J.; Clarke, Frank M.; Rogers, Peter J.; Young, Paul; Sales, Narelle; O’Doherty, Patrick J.; Higgins, Vincent J.

    2011-01-01

    This study was carried out with fresh Australian lager beer which was sampled directly off the production line, the same samples aged for 12 weeks at 30 °C, and the vintage beer which was kept at 20 °C for 5 years. Characteristic Australian lager flavour was maintained in the fresh and vintage beers but was lost in the aged beer. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and free thiol group labelling analyses of beer proteins found that this flavour stability correlated with the presence of an unknown 10 kilodaltons (kDa) protein with a higher level of free thiols. The protein was purified by size-exclusion chromatography, then peptide sequencing and database matching identified it as the barley lipid transfer protein (LTP1). Further characterisation using diphenylpicrylhydrazyl (DPPH) free radical scavenging and a Saccharomyces cerevisiae-based antioxidant screening assay demonstrated that the LTP1 protein was active in DPPH reduction and antioxidant activity. The absence of free thiol in the aged beer indicates that the thiol functional groups within the LTP1 protein were saturated and suggests that it is important in the flavour stability of beer by maintaining reduction capacity during the ageing process. PMID:22016646

  3. The chromodomains of CHD1 are critical for enzymatic activity but less important for chromatin localization.

    PubMe