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  1. High levels of anti-Nef antibodies may prevent AIDS disease progression in vertically HIV-1-infected infants

    PubMed Central

    Corró, Guillermo; Crudeli, Cintia Milena; Rocco, Carlos Alberto; Marino, Silvia Alejandra; Sen, Luisa

    2014-01-01

    Introduction HIV-1-associated CD4+ T-cell depletion is a consequence of uninfected cell death. Nef is one of the viral factors that trigger apoptosis on bystander cells, though the plasma Nef levels do not correlate with Th lymphocytes counts. The aim of our study was to evaluate whether anti-Nef antibodies were involved in paediatric AIDS development and whether they can prevent the CD4+ T-cell depletion in vertically infected children. Methods Two hundred and seventy three HIV-1 vertically infected children seen at Garrahan Paediatric Hospital were randomly included in the study, adding 13 selected cases: seven LTNP (long-term non-progressors) and six RP (rapid progressors) children (n total=286). Specific anti-HIV-1-Nef antibodies were titrated by indirect ELISA and compared between groups. The plasma blocking effect on Nef-dependent cytotoxicity was evaluated in Jurkat cells using recombinant Nef as apoptotic stimulus and patient plasmas as blockers, measuring the apoptotic levels using Annexin-V stain and flow cytometry. Results Only 63.4% of the patients had specific anti-Nef antibodies, and the levels of anti-Nef antibodies found in the selected LTNPs plasmas were always significantly higher (p=1.55×10−4) than those in RPs or general HIV-1+ paediatric populations. The LTNPs’ plasma had a strong inhibitory effect on Nef-dependent cytotoxicity even at high dilutions, while RP plasmas had little or no effect on Nef-induced apoptosis. Discussion and conclusions High anti-Nef antibody levels are associated and predict slow or non-progression to AIDS in vertically HIV-1-infected children. They could be an efficient tool in preventing Nef-associated bystander effect, preserving CD4+ T-cells and the immune function in the context of paediatric HIV-1 infection. PMID:24560340

  2. High-level HIV-1 Nef transient expression in Nicotiana benthamiana using the P19 gene silencing suppressor protein of Artichoke Mottled Crinckle Virus

    PubMed Central

    2009-01-01

    Background In recent years, different HIV antigens have been successfully expressed in plants by either stable transformation or transient expression systems. Among HIV proteins, Nef is considered a promising target for the formulation of a multi-component vaccine due to its implication in the first steps of viral infection. Attempts to express Nef as a single protein product (not fused to a stabilizing protein) in transgenic plants resulted in disappointingly low yields (about 0.5% of total soluble protein). In this work we describe a transient expression system based on co-agroinfiltration of plant virus gene silencing suppressor proteins in Nicotiana benthamiana, followed by a two-step affinity purification protocol of plant-derived Nef. Results The effect of three gene silencing viral suppressor proteins (P25 of Potato Virus X, P19 of either Artichoke Mottled Crinckle virus and Tomato Bushy Stunt virus) on Nef transient expression yield was evaluated. The P19 protein of Artichoke Mottled Crinckle virus (AMCV-P19) gave the highest expression yield in vacuum co-agroinfiltration experiments reaching 1.3% of total soluble protein, a level almost three times higher than that previously reported in stable transgenic plants. The high yield observed in the co-agroinfiltrated plants was correlated to a remarkable decrease of Nef-specific small interfering RNAs (siRNAs) indicating an effective modulation of RNA silencing mechanisms by AMCV-P19. Interestingly, we also showed that expression levels in top leaves of vacuum co-agroinfiltrated plants were noticeably reduced compared to bottom leaves. Moreover, purification of Nef from agroinfiltrated tissue was achieved by a two-step immobilized metal ion affinity chromatography protocol with yields of 250 ng/g of fresh tissue. Conclusion We demonstrated that expression level of HIV-1 Nef in plant can be improved using a transient expression system enhanced by the AMCV-P19 gene silencing suppressor protein. Moreover, plant

  3. Plant-based strategies aimed at expressing HIV antigens and neutralizing antibodies at high levels. Nef as a case study.

    PubMed

    Marusic, Carla; Vitale, Alessandro; Pedrazzini, Emanuela; Donini, Marcello; Frigerio, Lorenzo; Bock, Ralph; Dix, Philip J; McCabe, Matthew S; Bellucci, Michele; Benvenuto, Eugenio

    2009-08-01

    The first evidence that plants represent a valid, safe and cost-effective alternative to traditional expression systems for large-scale production of antigens and antibodies was described more than 10 years ago. Since then, considerable improvements have been made to increase the yield of plant-produced proteins. These include the use of signal sequences to target proteins to different cellular compartments, plastid transformation to achieve high transgene dosage, codon usage optimization to boost gene expression, and protein fusions to improve recombinant protein stability and accumulation. Thus, several HIV/SIV antigens and neutralizing anti-HIV antibodies have recently been successfully expressed in plants by stable nuclear or plastid transformation, and by transient expression systems based on plant virus vectors or Agrobacterium-mediated infection. The current article gives an overview of plant expressed HIV antigens and antibodies and provides an account of the use of different strategies aimed at increasing the expression of the accessory multifunctional HIV-1 Nef protein in transgenic plants.

  4. Identification of a highly conserved valine-glycine-phenylalanine amino acid triplet required for HIV-1 Nef function.

    PubMed

    Meuwissen, Pieter J; Stolp, Bettina; Iannucci, Veronica; Vermeire, Jolien; Naessens, Evelien; Saksela, Kalle; Geyer, Matthias; Vanham, Guido; Arien, Kevin K; Fackler, Oliver T; Verhasselt, Bruno

    2012-04-27

    The Nef protein of HIV facilitates virus replication and disease progression in infected patients. This role as pathogenesis factor depends on several genetically separable Nef functions that are mediated by interactions of highly conserved protein-protein interaction motifs with different host cell proteins. By studying the functionality of a series of nef alleles from clinical isolates, we identified a dysfunctional HIV group O Nef in which a highly conserved valine-glycine-phenylalanine (VGF) region, which links a preceding acidic cluster with the following proline-rich motif into an amphipathic surface was deleted. In this study, we aimed to study the functional importance of this VGF region. The dysfunctional HIV group O8 nef allele was restored to the consensus sequence, and mutants of canonical (NL4.3, NA-7, SF2) and non-canonical (B2 and C1422) HIV-1 group M nef alleles were generated in which the amino acids of the VGF region were changed into alanines (VGF→AAA) and tested for their capacity to interfere with surface receptor trafficking, signal transduction and enhancement of viral replication and infectivity. We found the VGF motif, and each individual amino acid of this motif, to be critical for downregulation of MHC-I and CXCR4. Moreover, Nef's association with the cellular p21-activated kinase 2 (PAK2), the resulting deregulation of cofilin and inhibition of host cell actin remodeling, and targeting of Lck kinase to the trans-golgi-network (TGN) were affected as well. Of particular interest, VGF integrity was essential for Nef-mediated enhancement of HIV virion infectivity and HIV replication in peripheral blood lymphocytes. For targeting of Lck kinase to the TGN and viral infectivity, especially the phenylalanine of the triplet was essential. At the molecular level, the VGF motif was required for the physical interaction of the adjacent proline-rich motif with Hck. Based on these findings, we propose that this highly conserved three amino acid VGF

  5. A Highly Conserved Residue in HIV-1 Nef Alpha Helix 2 Modulates Protein Expression

    PubMed Central

    Johnson, Aaron L.; Dirk, Brennan S.; Coutu, Mathieu; Haeryfar, S. M. Mansour; Arts, Eric J.; Finzi, Andrés

    2016-01-01

    ABSTRACT Extensive genetic diversity is a defining characteristic of human immunodeficiency virus type 1 (HIV-1) and poses a significant barrier to the development of an effective vaccine. To better understand the impact of this genetic diversity on the HIV-1 pathogenic factor Nef, we compiled a panel of reference strains from the NIH Los Alamos HIV Database. Initial sequence analysis identified point mutations at Nef residues 13, 84, and 92 in subtype C reference strain C.BR92025 from Brazil. Functional analysis revealed impaired major histocompatibility complex class I and CD4 downregulation of strain C.BR92025 Nef, which corresponded to decreased protein expression. Metabolic labeling demonstrated that strain C.BR92025 Nef has a greater rate of protein turnover than subtype B reference strain B.JRFL that, on the basis of mutational analysis, is related to Nef residue A84. An alanine-to-valine substitution at position 84, located in alpha helix 2 of Nef, was sufficient to alter the rate of turnover of an otherwise highly expressed Nef protein. In conclusion, these findings highlight HIV-1 Nef residue A84 as a major determinant of protein expression that may offer an additional avenue to disrupt or mediate the effects of this key HIV-1 pathogenic factor. IMPORTANCE The HIV-1 Nef protein has been established as a key pathogenic determinant of HIV/AIDS, but there is little knowledge of how the extensive genetic diversity of HIV-1 affects Nef function. Upon compiling a set of subtype-specific reference strains, we identified a subtype C reference strain, C.BR92025, that contained natural polymorphisms at otherwise highly conserved residues 13, 84, and 92. Interestingly, strain C.BR92025 Nef displayed impaired Nef function and had decreased protein expression. We have demonstrated that strain C.BR92025 Nef has a higher rate of protein turnover than highly expressed Nef proteins and that this higher rate of protein turnover is due to an alanine-to-valine substitution

  6. Identification of a highly conserved valine-glycine-phenylalanine amino acid triplet required for HIV-1 Nef function

    PubMed Central

    2012-01-01

    Background The Nef protein of HIV facilitates virus replication and disease progression in infected patients. This role as pathogenesis factor depends on several genetically separable Nef functions that are mediated by interactions of highly conserved protein-protein interaction motifs with different host cell proteins. By studying the functionality of a series of nef alleles from clinical isolates, we identified a dysfunctional HIV group O Nef in which a highly conserved valine-glycine-phenylalanine (VGF) region, which links a preceding acidic cluster with the following proline-rich motif into an amphipathic surface was deleted. In this study, we aimed to study the functional importance of this VGF region. Results The dysfunctional HIV group O8 nef allele was restored to the consensus sequence, and mutants of canonical (NL4.3, NA-7, SF2) and non-canonical (B2 and C1422) HIV-1 group M nef alleles were generated in which the amino acids of the VGF region were changed into alanines (VGF→AAA) and tested for their capacity to interfere with surface receptor trafficking, signal transduction and enhancement of viral replication and infectivity. We found the VGF motif, and each individual amino acid of this motif, to be critical for downregulation of MHC-I and CXCR4. Moreover, Nef’s association with the cellular p21-activated kinase 2 (PAK2), the resulting deregulation of cofilin and inhibition of host cell actin remodeling, and targeting of Lck kinase to the trans-golgi-network (TGN) were affected as well. Of particular interest, VGF integrity was essential for Nef-mediated enhancement of HIV virion infectivity and HIV replication in peripheral blood lymphocytes. For targeting of Lck kinase to the TGN and viral infectivity, especially the phenylalanine of the triplet was essential. At the molecular level, the VGF motif was required for the physical interaction of the adjacent proline-rich motif with Hck. Conclusion Based on these findings, we propose that this highly

  7. HIV Type 1 Nef Is Released from Infected Cells in CD45+ Microvesicles and Is Present in the Plasma of HIV-Infected Individuals

    PubMed Central

    Raymond, A.D.; Campbell-Sims, T.C.; Khan, M.; Lang, M.; Huang, M.B.; Bond, V.C.

    2011-01-01

    Abstract HIV-1 Nef has been demonstrated to be integral for viral persistence, infectivity, and the acceleration of disease pathogenesis (AIDS) in humans. Nef has also been detected in the plasma of HIV-infected individuals and is released from infected cells. The form in which Nef is released from infected cells is unknown. However, Nef is a myristoylated protein and has been shown to interact with the intracellular vesicular trafficking network. Here we show that Nef is released in CD45-containing microvesicles. This microvesicular Nef (mvNef) is detected in the plasma of HIV-infected individuals at relatively high concentrations (10 ng/ml). It is also present in tissue culture supernatants of Jurkat cells infected with HIVMN. Interestingly, plasma mvNef levels in HIV+ patients did not significantly correlate with viral load or CD4 count. Microvesicular Nef levels persisted in the plasma of HIV-infected individuals despite the use of antiretroviral therapy, even in individuals with undetectable viral loads. Using cell lines, we found Nef microvesicles induce apoptosis in Jurkat T-lymphocytes but had no observed effect on the U937 monocytic cell line. Given the large amount of mvNef present in the plasma of HIV-infected individuals, the apoptotic effect of mvNef on T cells, and the observed functions of extracellular soluble Nef in vitro, it seems likely that in vivo mvNef may play a significant role in the pathogenesis of AIDS. PMID:20964480

  8. Differential Ability of Primary HIV-1 Nef Isolates To Downregulate HIV-1 Entry Receptors

    PubMed Central

    Toyoda, Mako; Ogata, Yoko; Mahiti, Macdonald; Maeda, Yosuke; Kuang, Xiaomei T.; Miura, Toshiyuki; Jessen, Heiko; Walker, Bruce D.; Brockman, Mark A.; Brumme, Zabrina L.

    2015-01-01

    ABSTRACT HIV-1 Nef downregulates the viral entry receptor CD4 as well as the coreceptors CCR5 and CXCR4 from the surface of HIV-infected cells, and this leads to promotion of viral replication through superinfection resistance and other mechanisms. Nef sequence motifs that modulate these functions have been identified via in vitro mutagenesis with laboratory HIV-1 strains. However, it remains unclear whether the same motifs contribute to Nef activity in patient-derived sequences and whether these motifs may differ in Nef sequences isolated at different infection stages and/or from patients with different disease phenotypes. Here, nef clones from 45 elite controllers (EC), 46 chronic progressors (CP), and 43 acute progressors (AP) were examined for their CD4, CCR5, and CXCR4 downregulation functions. Nef clones from EC exhibited statistically significantly impaired CD4 and CCR5 downregulation ability and modestly impaired CXCR4 downregulation activity compared to those from CP and AP. Nef's ability to downregulate CD4 and CCR5 correlated positively in all cohorts, suggesting that they are functionally linked in vivo. Moreover, impairments in Nef's receptor downregulation functions increased the susceptibility of Nef-expressing cells to HIV-1 infection. Mutagenesis studies on three functionally impaired EC Nef clones revealed that multiple residues, including those at novel sites, were involved in the alteration of Nef functions and steady-state protein levels. Specifically, polymorphisms at highly conserved tryptophan residues (e.g., Trp-57 and Trp-183) and immune escape-associated sites were responsible for reduced Nef functions in these clones. Our results suggest that the functional modulation of primary Nef sequences is mediated by complex polymorphism networks. IMPORTANCE HIV-1 Nef, a key factor for viral pathogenesis, downregulates functionally important molecules from the surface of infected cells, including the viral entry receptor CD4 and coreceptors CCR5

  9. The human immunodeficiency virus-1 nef gene product: a positive factor for viral infection and replication in primary lymphocytes and macrophages

    PubMed Central

    1994-01-01

    Considerable controversy and uncertainty have surrounded the biological function of the Human Immunodeficiency Virus (HIV)-1 nef gene product. Initial studies suggested that this early, nonstructural viral protein functioned as a negative regulatory factor; thus, it was proposed to play a role in establishing or maintaining viral latency. In contrast, studies in Simian Immunodeficiency Virus (SIV)mac-infected rhesus monkeys have suggested that Nef is not a negative factor but rather plays a central role in promoting high-level viral replication and is required for viral pathogenesis in vivo. We sought to define a tissue culture system that would approximate the in vivo setting for virus infection in order to assess the role of HIV-1 Nef in viral replication. We show that infection of mitogen-activated peripheral blood mononuclear cells (PBMC) with Nef+ HIV results in enhanced replication as evidenced by earlier gag p24 expression when compared with infections performed with nef mutant viruses. Moreover, when unstimulated freshly isolated PBMC are infected with Nef+ and Nef- viruses and then subsequently activated with mitogen, the Nef-induced difference in viral replication kinetics is even more pronounced, with the Nef- viruses requiring much more time in culture for appreciable growth. A positive effect of Nef on viral replication was also observed in primary macrophages infected with a recombinant of YU-2, a patient- derived molecular clone with macrophage tropism. These positive effects of Nef on viral replication are dependent on the initial multiplicity of infection (MOI), in that infections of unstimulated PBMC at low MOI are most dependent upon intact nef for subsequent viral growth. We now provide evidence that the Nef+ HIV is more infectious than Nef- HIV from both a tissue culture infectious dose analysis, and a single-cell HIV infection assay. In the latter case, we demonstrate that infection with equivalent doses of HIV based on virion-associated gag p

  10. Efficient Nef-Mediated Downmodulation of TCR-CD3 and CD28 Is Associated with High CD4+ T Cell Counts in Viremic HIV-2 Infection

    PubMed Central

    Yu, Hangxing; Sauter, Daniel; Usmani, Shariq M.; Schmokel, Jan; Feldman, Jerome; Gruters, Rob A.; van der Ende, Marchina E.; Geyer, Matthias; Rowland-Jones, Sarah; Osterhaus, Albert D.

    2012-01-01

    The role of the multifunctional accessory Nef protein in the immunopathogenesis of HIV-2 infection is currently poorly understood. Here, we performed comprehensive functional analyses of 50 nef genes from 21 viremic (plasma viral load, >500 copies/ml) and 16 nonviremic (<500) HIV-2-infected individuals. On average, nef alleles from both groups were equally active in modulating CD4, TCR-CD3, CD28, MHC-I, and Ii cell surface expression and in enhancing virion infectivity. Thus, many HIV-2-infected individuals efficiently control the virus in spite of efficient Nef function. However, the potency of nef alleles in downmodulating TCR-CD3 and CD28 to suppress the activation and apoptosis of T cells correlated with high numbers of CD4+ T cells in viremic patients. No such correlations were observed in HIV-2-infected individuals with undetectable viral load. Further functional analyses showed that the Nef-mediated downmodulation of TCR-CD3 suppressed the induction of Fas, Fas-L, PD-1, and CTLA-4 cell surface expression as well as the secretion of gamma interferon (IFN-γ) by primary CD4+ T cells. Moreover, we identified a single naturally occurring amino acid variation (I132T) in the core domain of HIV-2 Nef that selectively disrupts its ability to downmodulate TCR-CD3 and results in functional properties highly reminiscent of HIV-1 Nef proteins. Taken together, our data suggest that the efficient Nef-mediated downmodulation of TCR-CD3 and CD28 help viremic HIV-2-infected individuals to maintain normal CD4+ T cell homeostasis by preventing T cell activation and by suppressing the induction of death receptors that may affect the functionality and survival of both virally infected and uninfected bystander cells. PMID:22345473

  11. Efficient Nef-mediated downmodulation of TCR-CD3 and CD28 is associated with high CD4+ T cell counts in viremic HIV-2 infection.

    PubMed

    Khalid, Mohammad; Yu, Hangxing; Sauter, Daniel; Usmani, Shariq M; Schmokel, Jan; Feldman, Jerome; Gruters, Rob A; van der Ende, Marchina E; Geyer, Matthias; Rowland-Jones, Sarah; Osterhaus, Albert D; Kirchhoff, Frank

    2012-05-01

    The role of the multifunctional accessory Nef protein in the immunopathogenesis of HIV-2 infection is currently poorly understood. Here, we performed comprehensive functional analyses of 50 nef genes from 21 viremic (plasma viral load, >500 copies/ml) and 16 nonviremic (<500) HIV-2-infected individuals. On average, nef alleles from both groups were equally active in modulating CD4, TCR-CD3, CD28, MHC-I, and Ii cell surface expression and in enhancing virion infectivity. Thus, many HIV-2-infected individuals efficiently control the virus in spite of efficient Nef function. However, the potency of nef alleles in downmodulating TCR-CD3 and CD28 to suppress the activation and apoptosis of T cells correlated with high numbers of CD4(+) T cells in viremic patients. No such correlations were observed in HIV-2-infected individuals with undetectable viral load. Further functional analyses showed that the Nef-mediated downmodulation of TCR-CD3 suppressed the induction of Fas, Fas-L, PD-1, and CTLA-4 cell surface expression as well as the secretion of gamma interferon (IFN-γ) by primary CD4(+) T cells. Moreover, we identified a single naturally occurring amino acid variation (I132T) in the core domain of HIV-2 Nef that selectively disrupts its ability to downmodulate TCR-CD3 and results in functional properties highly reminiscent of HIV-1 Nef proteins. Taken together, our data suggest that the efficient Nef-mediated downmodulation of TCR-CD3 and CD28 help viremic HIV-2-infected individuals to maintain normal CD4(+) T cell homeostasis by preventing T cell activation and by suppressing the induction of death receptors that may affect the functionality and survival of both virally infected and uninfected bystander cells.

  12. HIV-1 Nef protein in the nucleus influences adipogenesis as well as viral transcription through the peroxisome proliferator-activated receptors.

    PubMed

    Otake, Kaori; Omoto, Shinya; Yamamoto, Takuya; Okuyama, Harumi; Okada, Hidechika; Okada, Noriko; Kawai, Masahiro; Saksena, Nitin K; Fujii, Yoichi R

    2004-01-23

    Although the HIV-1 Nef protein (27 kDa) localizes primarily in cytoplasm, there is considerable evidence suggesting its occasional localization in the nucleus. Nef is known to play an important role in transcriptional events and viral replication, but the actual target of Nef in the nucleus remains to be identified. To examine the functional roles of Nef in the nucleus and its possible interactions with other unknown factors in the nucleus. High-density microarray analysis was used to screen directly the unique functions of Nef on host gene transcription. The nuclear localization of Nef and its effects on the expression of peroxisome proliferator-activated receptors (PPAR) was examined using PPAR promoter/reporter assay and immunoblotting. A long terminal repeat/reporter assay was used to investigated the effects of Nef and PPAR on viral transcription. Nef in the nucleus suppressed PPAR gamma expression and reduced fatty acid levels in human T and macrophage cell lines. Expression of Nef or PPAR suppressed viral replication; the effect of PPAR gamma or retinoid X receptor-alpha on viral replication were reduced by coexpression of Nef in MT(-)4 T cells. Nef may be involved in both viral replication and the wasting syndrome associated with AIDS.

  13. HIV protein Nef causes dyslipidemia and formation of foam cells in mouse models of atherosclerosis

    PubMed Central

    Cui, Huanhuan L.; Ditiatkovski, Michael; Kesani, Rajitha; Bobryshev, Yuri V.; Liu, Yingying; Geyer, Matthias; Mukhamedova, Nigora; Bukrinsky, Michael; Sviridov, Dmitri

    2014-01-01

    Patients with HIV are at an increased risk of cardiovascular disease. In this study we investigated the effect of Nef, a secreted HIV protein responsible for the impairment of cholesterol efflux, on the development of atherosclerosis in two animal models. ApoE−/− mice fed a high-fat diet and C57BL/6 mice fed a high-fat, high-cholesterol diet were injected with recombinant Nef (40 ng/injection) or vehicle, and the effects of Nef on development of atherosclerosis, inflammation, and dyslipidemia were assessed. In apoE−/− mice, Nef significantly increased the size of atherosclerotic lesions and caused vessel remodeling. Nef caused elevation of total cholesterol and triglyceride levels in the plasma while reducing high-density lipoprotein cholesterol levels. These changes were accompanied by a reduction of ABCA1 abundance in the liver, but not in the vessels. In C57BL/6 mice, Nef caused a significant number of lipid-laden macrophages presented in adventitia of the vessels; these cells were absent from the vessels of control mice. Nef caused sharp elevations of plasma triglyceride levels and body weight. Taken together, our findings suggest that Nef causes dyslipidemia and accumulation of cholesterol in macrophages within the vessel wall, supporting the role of Nef in pathogenesis of atherosclerosis in HIV-infected patients.—Cui, H. L., Ditiatkovski, M., Kesani, R., Bobryshev, Y. V., Liu, Y., Geyer, M., Mukhamedova, N., Bukrinsky, M., Sviridov, D. HIV protein Nef causes dyslipidemia and formation of foam cells in mouse models of atherosclerosis. PMID:24642731

  14. Microglia-derived HIV Nef+ exosome impairment of the blood-brain barrier is treatable by nanomedicine-based delivery of Nef peptides.

    PubMed

    Raymond, A D; Diaz, P; Chevelon, S; Agudelo, M; Yndart-Arias, A; Ding, H; Kaushik, A; Jayant, R Dev; Nikkhah-Moshaie, R; Roy, U; Pilakka-Kanthikeel, S; Nair, M P

    2016-04-01

    The negative factor (Nef) of human immunodeficiency virus (HIV) is an accessory protein that is thought to be integral to HIV-associated immune- and neuroimmune pathogenesis. Here, we show that nef-transfected microglia-released Nef+ exosome (exNef) disrupts the apical blood-brain barrier (BBB) and that only nef-transfected microglia release Nef in exosomes. nef-gfp-transduced neurons and astrocytes release exosomes but did not release exNef in the extracellular space. Apical administration of exNef derived from nef-transfected 293T cells reduced transendothelial electrical resistance (TEER) and increased permeability of the BBB. Microglia-derived exNef applied to either the apical/basal BBB significantly reduced expression of the tight junction protein, ZO-1, suggesting a mechanism of exNef-mediated neuropathogenesis. Microglia exposed to exNef release elevated levels of Toll-like receptor-induced cytokines and chemokines IL-12, IL-8, IL-6, RANTES, and IL-17A. Magnetic nanoparticle delivery of Nef peptides containing the Nef myrisolation site across an in vitro BBB ultimately reduced nef-transfected microglia release of Nef exosomes and prevented the loss of BBB integrity and permeability as measured by TEER and dextran-FITC transport studies, respectively. Overall, we show that exNef is released from nef-gfp-transfected microglia; exNef disrupts integrity and permeability, and tight junctions of the BBB, and induces microglial cytokine/chemokine secretion. These exNef-mediated effects were significantly restricted by Nef peptides. Taken together, this study provides preliminary evidence of the role of exNef in HIV neuroimmune pathogenesis and the feasibility of a nanomedicine-based therapeutics targeting exNef to treat HIV-associated neuropathogenesis.

  15. Viremic long-term nonprogressive HIV-1 infection is not associated with abnormalities in known Nef functions.

    PubMed

    Heigele, Anke; Camerini, David; van't Wout, Angélique B; Kirchhoff, Frank

    2014-02-04

    A small minority of HIV-1-infected individuals show low levels of immune activation and do not develop immunodeficiency despite high viral loads. Since the accessory viral Nef protein modulates T cell activation and plays a key role in the pathogenesis of AIDS, we investigated whether specific properties of Nef may be associated with this highly unusual clinical outcome of HIV-1 infection. Comprehensive functional analyses of sequential HIV-1 strains from three viremic long-term non-progressors (VNP) showed that they encode full-length Nef proteins that are capable of modulating CD4, CD28, CD8ß, MHC-I and CD74 cell surface expression. Similar to Nef proteins from HIV-1-infected individuals with progressive infection (P-Nefs) and unlike Nefs from simian immunodeficiency viruses (SIVs) that do not cause chronic immune activation and disease in their natural simian hosts, VNP-Nefs were generally unable to down-modulate TCR-CD3 cell surface expression to block T cell activation and apoptosis. On average, VNP-Nefs suppressed NF-AT activation less effectively than P-Nefs and were slightly less active in enhancing NF-κB activity. Finally, we found that VNP-Nefs increased virion infectivity and enhanced HIV-1 replication and cytopathicity in primary human cells and in ex vivo infected lymphoid tissues. Our results show that nef alleles from VNPs and progressors of HIV-1 infection show only modest differences in established functions. Thus, the lack of chronic immune activation and disease progression in HIV-1-infected VNPs is apparently not associated with unusual functional properties of the accessory viral Nef protein.

  16. A Conserved GPG-Motif in the HIV-1 Nef Core Is Required for Principal Nef-Activities

    PubMed Central

    Martínez-Bonet, Marta; Palladino, Claudia; Briz, Veronica; Rudolph, Jochen M.; Fackler, Oliver T.; Relloso, Miguel; Muñoz-Fernandez, Maria Angeles; Madrid, Ricardo

    2015-01-01

    To find out new determinants required for Nef activity we performed a functional alanine scanning analysis along a discrete but highly conserved region at the core of HIV-1 Nef. We identified the GPG-motif, located at the 121–137 region of HIV-1 NL4.3 Nef, as a novel protein signature strictly required for the p56Lck dependent Nef-induced CD4-downregulation in T-cells. Since the Nef-GPG motif was dispensable for CD4-downregulation in HeLa-CD4 cells, Nef/AP-1 interaction and Nef-dependent effects on Tf-R trafficking, the observed effects on CD4 downregulation cannot be attributed to structure constraints or to alterations on general protein trafficking. Besides, we found that the GPG-motif was also required for Nef-dependent inhibition of ring actin re-organization upon TCR triggering and MHCI downregulation, suggesting that the GPG-motif could actively cooperate with the Nef PxxP motif for these HIV-1 Nef-related effects. Finally, we observed that the Nef-GPG motif was required for optimal infectivity of those viruses produced in T-cells. According to these findings, we propose the conserved GPG-motif in HIV-1 Nef as functional region required for HIV-1 infectivity and therefore with a potential interest for the interference of Nef activity during HIV-1 infection. PMID:26700863

  17. Replication of HIV-1 deleted Nef mutants in chronically immune activated human T cells.

    PubMed

    Shapira-Nahor, Orit; Maayan, Shlomo; Peden, Keith W C; Rabinowitz, Ruth; Schlesinger, Michael; Alian, Akram; Panet, Amos

    2002-11-10

    Lymphocytes (PBMC) obtained from blood of HIV-sera negative Ethiopian immigrants (ETH) were highly susceptible to HIV-1 infection in vitro with no need for stimulation by mitogens. As the HIV nef gene product has been shown to enhance viral replication in stimulated primary lymphocytes, we investigated in this work the role of Nef in viral replication in the ETH cells. Lymphocytes obtained from ETH individuals supported high replication of wild-type HIV-1 and low but significant replication level of the two deleted Nef mutants (encode truncated Nef proteins consisting only of either the first 35 or the first 86 amino acids of Nef). In contrast, no replication was observed in nonactivated cells obtained from non-ETH individuals. After activation of the PBMC from ETH individuals with PHA, replication of both wild-type strains and the two deleted Nef mutant viruses further increased. The CD4(+) T cells of ETH individuals exhibited elevated levels of the surface activation markers CD45RO and HLA-DR, compared with T cells derived from non-ETH group. Likewise, expression of the chemokine receptors CCR5 and CXCR4 on these cells was higher in the ETH group than in the non-ETH group. Replication of HIV-1 wild-type and the isogenic-deleted Nef mutants was significantly correlated with the proportion of ETH cells expressing CD45RO and the chemokine receptors. This study suggests that HIV-1 may respond differently to several activation states characteristic of T cells. One activation state, defined by chronically activated lymphocytes from ETH individuals, is permissive to the wild-type HIV-1 and, to a lesser degree, to the Nef mutants. Further activation of these cells by exogenous stimuli enhances replication of the virus. Our results support the notion that Nef enhances the basal level of T cell activation and consequently, viral replication.

  18. cis Expression of the F12 Human Immunodeficiency Virus (HIV) Nef Allele Transforms the Highly Productive NL4-3 HIV Type 1 to a Replication-Defective Strain: Involvement of both Env gp41 and CD4 Intracytoplasmic Tails

    PubMed Central

    Olivetta, Eleonora; Pugliese, Katherina; Bona, Roberta; D'Aloja, Paola; Ferrantelli, Flavia; Santarcangelo, Anna Claudia; Mattia, Gianfranco; Verani, Paola; Federico, Maurizio

    2000-01-01

    F12 human immunodeficiency virus type 1 (HIV-1) nef is a naturally occurring nef mutant cloned from the provirus of a nonproductive, nondefective, and interfering HIV-1 variant (F12-HIV). We have already shown that cells stably transfected with a vector expressing the F12-HIV nef allele do not downregulate CD4 receptors and, more peculiarly, become resistant to the replication of wild type (wt) HIV. In order to investigate the mechanism of action of such an HIV inhibition, the F12-HIV nef gene was expressed in the context of the NL4-3 HIV-1 infectious molecular clone by replacing the wt nef gene (NL4-3/chi). Through this experimental approach we established the following. First, NL4-3/chi and nef-defective (Δnef) NL4-3 viral particles behave very similarly in terms of viral entry and HIV protein production during the first replicative cycle. Second, no viral particles were produced from cells infected with NL4-3/chi virions, whatever the multiplicity of infection used. The viral inhibition apparently occurs at level of viral assembling and/or release. Third, this block could not be relieved by in-trans expression of wt nef. Finally, NL4-3/chi reverts to a producer HIV strain when F12-HIV Nef is deprived of its myristoyl residue. Through a CD4 downregulation competition assay, we demonstrated that F12-HIV Nef protein potently inhibits the CD4 downregulation induced by wt Nef. Moreover, we observed a redistribution of CD4 receptors at the cell margin induced by F12-HIV Nef. These observations strongly suggest that F12-HIV Nef maintains the ability to interact with the intracytoplasmic tail of the CD4 receptor molecule. Remarkably, we distinguished the intracytoplasmic tails of Env gp41 and CD4 as, respectively, viral and cellular targets of the F12-HIV Nef-induced viral retention. For the first time, the inhibition of the viral life cycle by means of in-cis expression of a Nef mutant is here reported. Delineation of the F12-HIV Nef mechanism of action may offer

  19. Secretion modification region-derived peptide disrupts HIV-1 Nef's interaction with mortalin and blocks virus and Nef exosome release.

    PubMed

    Shelton, Martin N; Huang, Ming-Bo; Ali, Syed A; Powell, Michael D; Bond, Vincent C

    2012-01-01

    Nef is secreted from infected cells in exosomes and is found in abundance in the sera of HIV-infected individuals. Secreted exosomal Nef (exNef) induces apoptosis in uninfected CD4⁺ T cells and may be a key component of HIV pathogenesis. The exosomal pathway has been implicated in HIV-1 virus release, suggesting a possible link between these two viral processes. However, the underlying mechanisms and cellular components of exNef secretion have not been elucidated. We have previously described a Nef motif, the secretion modification region (SMR; amino acids 66 to 70), that is required for exNef secretion. In silico modeling data suggest that this motif can form a putative binding pocket. We hypothesized that the Nef SMR binds a cellular protein involved in protein trafficking and that inhibition of this interaction would abrogate exNef secretion. By using tandem mass spectrometry and coimmunoprecipitation with a novel SMR-based peptide (SMRwt) that blocks exNef secretion and HIV-1 virus release, we identified mortalin as an SMR-specific cellular protein. A second set of coimmunoprecipitation experiments with full-length Nef confirmed that mortalin interacts with Nef via Nef's SMR motif and that this interaction is disrupted by the SMRwt peptide. Overexpression and microRNA knockdown of mortalin revealed a positive correlation between exNef secretion levels and mortalin protein expression. Using antibody inhibition we demonstrated that the Nef/mortalin interaction is necessary for exNef secretion. Taken together, this work constitutes a significant step in understanding the underlying mechanism of exNef secretion, identifies a novel host-pathogen interaction, and introduces an HIV-derived peptide with antiviral properties.

  20. Persistent Low-Level Replication of SIVΔnef Drives Maturation of Antibody and CD8 T Cell Responses to Induce Protective Immunity against Vaginal SIV Infection

    PubMed Central

    Gillis, Jacqueline; Wong, Fay E.; Yu, Yi; Camp, Jeremy V.; Li, Qingsheng; Connole, Michelle; Li, Yuan; Lifson, Jeffrey D.; Li, Wenjun; Keele, Brandon F.; Kozlowski, Pamela A.; Desrosiers, Ronald C.; Haase, Ashley T.

    2016-01-01

    Defining the correlates of immune protection conferred by SIVΔnef, the most effective vaccine against SIV challenge, could enable the design of a protective vaccine against HIV infection. Here we provide a comprehensive assessment of immune responses that protect against SIV infection through detailed analyses of cellular and humoral immune responses in the blood and tissues of rhesus macaques vaccinated with SIVΔnef and then vaginally challenged with wild-type SIV. Despite the presence of robust cellular immune responses, animals at 5 weeks after vaccination displayed only transient viral suppression of challenge virus, whereas all macaques challenged at weeks 20 and 40 post-SIVΔnef vaccination were protected, as defined by either apparent sterile protection or significant suppression of viremia in infected animals. Multiple parameters of CD8 T cell function temporally correlated with maturation of protection, including polyfunctionality, phenotypic differentiation, and redistribution to gut and lymphoid tissues. Importantly, we also demonstrate the induction of a tissue-resident memory population of SIV-specific CD8 T cells in the vaginal mucosa, which was dependent on ongoing low-level antigenic stimulation. Moreover, we show that vaginal and serum antibody titers inversely correlated with post-challenge peak viral load, and we correlate the accumulation and affinity maturation of the antibody response to the duration of the vaccination period as well as to the SIVΔnef antigenic load. In conclusion, maturation of SIVΔnef-induced CD8 T cell and antibody responses, both propelled by viral persistence in the gut mucosa and secondary lymphoid tissues, results in protective immune responses that are able to interrupt viral transmission at mucosal portals of entry as well as potential sites of viral dissemination. PMID:27959961

  1. Analysis of human immunodeficiency virus type 1 nef gene sequences present in vivo.

    PubMed Central

    Shugars, D C; Smith, M S; Glueck, D H; Nantermet, P V; Seillier-Moiseiwitsch, F; Swanstrom, R

    1993-01-01

    The nef genes of the human immunodeficiency viruses type 1 and 2 (HIV-1 and HIV-2) and the related simian immunodeficiency viruses (SIVs) encode a protein (Nef) whose role in virus replication and cytopathicity remains uncertain. As an attempt to elucidate the function of nef, we characterized the nucleotide and corresponding protein sequences of naturally occurring nef genes obtained from several HIV-1-infected individuals. A consensus Nef sequence was derived and used to identify several features that were highly conserved among the Nef sequences. These features included a nearly invariant myristylation signal, regions of sequence polymorphism and variable duplication, a region with an acidic charge, a (Pxx)4 repeat sequence, and a potential protein kinase C phosphorylation site. Clustering of premature stop codons at position 124 was noted in 6 of the 54 Nef sequences. Further analysis revealed four stretches of residues that were highly conserved not only among the patient-derived HIV-1 Nef sequences, but also among the Nef sequences of HIV-2 and the SIVs, suggesting that Nef proteins expressed by these retroviruses are functionally equivalent. The "Nef-defining" sequences were used to evaluate the sequence alignments of known proteins reported to share sequence similarity with Nef sequences and to conduct additional computer-based searches for similar protein sequences. A gene encoding the consensus Nef sequence was also generated. This gene encodes a full-length Nef protein that should be a valuable tool in further studies of Nef function. Images PMID:8043040

  2. HIV-1 Nef Impairs Key Functional Activities in Human Macrophages through CD36 Downregulation

    PubMed Central

    Olivetta, Eleonora; Tirelli, Valentina; Chiozzini, Chiara; Scazzocchio, Beatrice; Romano, Ignazio; Arenaccio, Claudia; Sanchez, Massimo

    2014-01-01

    Monocytes and macrophages utilize the class A and B scavenger receptors to recognize and perform phagocytosis of invading microbes before a pathogen-specific immune response is generated. HIV-1 Nef protein affects the innate immune system impairing oxidative burst response and phagocytic capacity of macrophages. Our data show that exogenous recombinant myristoylated Nef protein induces a marked CD36 downregulation in monocytes from Peripheral Blood Mononuclear Cells, in Monocyte-Derived Macrophages (MDMs) differentiated by cytokines and in MDMs contained in a mixed culture obtained expanding PBMCs under Human Erythroid Massive Amplification condition. Under the latter culture condition we identify three main populations after 6 days of expansion: lymphocytes (37.8±14.7%), erythroblasts (46.7±6.1%) and MDMs (15.7±7.5%). The Nef addition to the cell culture significantly downregulates CD36 expression in MDMs, but not in erythroid cells. Furthermore, CD36 inhibition is highly specific since it does not modify the expression levels of other MDM markers such as CD14, CD11c, CD86, CD68, CD206, Toll-like Receptor 2 and Toll-like Receptor 4. Similar results were obtained in MDMs infected with VSV-G pseudotyped HIV-1-expressing Nef. The reduced CD36 membrane expression is associated with decrease of correspondent CD36 mRNA transcript. Furthermore, Nef-induced CD36 downregulation is linked to both impaired scavenger activity with reduced capability to take up oxidized lipoproteins and to significant decreased phagocytosis of fluorescent beads and GFP-expressing Salmonella tiphymurium. In addition we observed that Nef induces TNF-α release in MDMs. Although these data suggest a possible involvement of TNF-α in mediating Nef activity, our results exclude a possible relationship between Nef-induced TNF-α release and Nef-mediated CD36 downregulation. The present work shows that HIV-1 Nef protein may have a role in the strategies elaborated by HIV-1 to alter pathogen

  3. A Truncated Nef Peptide from SIVcpz Inhibits the Production of HIV-1 Infectious Progeny

    PubMed Central

    Sabino Cunha, Marcela; Lima Sampaio, Thatiane; Peterlin, B. Matija; Jesus da Costa, Luciana

    2016-01-01

    Nef proteins from all primate Lentiviruses, including the simian immunodeficiency virus of chimpanzees (SIVcpz), increase viral progeny infectivity. However, the function of Nef involved with the increase in viral infectivity is still not completely understood. Nonetheless, until now, studies investigating the functions of Nef from SIVcpz have been conducted in the context of the HIV-1 proviruses. In an attempt to investigate the role played by Nef during the replication cycle of an SIVcpz, a Nef-defective derivative was obtained from the SIVcpzWTGab2 clone by introducing a frame shift mutation at a unique restriction site within the nef sequence. This nef-deleted clone expresses an N-terminal 74-amino acid truncated peptide of Nef and was named SIVcpz-tNef. We found that the SIVcpz-tNef does not behave as a classic nef-deleted HIV-1 or simian immunodeficiency virus of macaques SIVmac. Markedly, SIVcpz-tNef progeny from both Hek-293T and Molt producer cells were completely non-infectious. Moreover, the loss in infectivity of SIVcpz-tNef correlated with the inhibition of Gag and GagPol processing. A marked accumulation of Gag and very low levels of reverse transcriptase were detected in viral lysates. Furthermore, these observations were reproduced once the tNef peptide was expressed in trans both in SIVcpzΔNef and HIV-1WT expressing cells, demonstrating that the truncated peptide is a dominant negative for viral processing and infectivity for both SIVcpz and HIV-1. We demonstrated that the truncated Nef peptide binds to GagPol outside the protease region and by doing so probably blocks processing of both GagPol and Gag precursors at a very early stage. This study demonstrates for the first time that naturally-occurring Nef peptides can potently block lentiviral processing and infectivity. PMID:27399760

  4. Development and validation of a high-content bimolecular fluorescence complementation assay for small-molecule inhibitors of HIV-1 Nef dimerization.

    PubMed

    Poe, Jerrod A; Vollmer, Laura; Vogt, Andreas; Smithgall, Thomas E

    2014-04-01

    Nef is a human immunodeficiency virus 1 (HIV-1) accessory factor essential for viral pathogenesis and AIDS progression. Many Nef functions require dimerization, and small molecules that block Nef dimerization may represent antiretroviral drug leads. Here we describe a cell-based assay for Nef dimerization inhibitors based on bimolecular fluorescence complementation (BiFC). Nef was fused to nonfluorescent, complementary fragments of yellow fluorescent protein (YFP) and coexpressed in the same cell population. Dimerization of Nef resulted in juxtaposition of the YFP fragments and reconstitution of the fluorophore. For automation, the Nef-YFP fusion proteins plus a monomeric red fluorescent protein (mRFP) reporter were expressed from a single vector, separated by picornavirus "2A" linker peptides to permit equivalent translation of all three proteins. Validation studies revealed a critical role for gating on the mRFP-positive subpopulation of transfected cells, as well as use of the mRFP signal to normalize the Nef-BiFC signal. Nef-BiFC/mRFP ratios resulting from cells expressing wild-type versus dimerization-defective Nef were very clearly separated, with Z factors consistently in the 0.6 to 0.7 range. A fully automated pilot screen of the National Cancer Institute Diversity Set III identified several hit compounds that reproducibly blocked Nef dimerization in the low micromolar range. This BiFC-based assay has the potential to identify cell-active small molecules that directly interfere with Nef dimerization and function.

  5. Suppression of HIV-1 Nef translation by Sam68 mutant-induced stress granules and nef mRNA sequestration

    PubMed Central

    Henao-Mejia, Jorge; Liu, Ying; Park, In-Woo; Zhang, Jizhong; Sanford, Jeremy; He, Johnny J.

    2009-01-01

    SUMMARY HIV-1 Nef plays important roles in HIV-1 replication and pathogenesis. It is translated from completely spliced HIV-1 RNA, its expression is inherently regulated at the levels of viral DNA transcription and RNA splicing. Here we show that Sam68 cytoplasmic mutants potently suppress Nef expression. The suppression requires Sam68 domain aa269-321 and is correlated with its ability to induce stress granules. In addition, the suppression is specific to Nef, and direct binding to nef mRNA 3′UTR confers the suppression specificity. Furthermore, nef mRNA is targeted to and enriched in these induced stress granules. Importantly, Nef suppression occurs in the context of HIV-1 infection of CD4+ T lymphocytes with little MHC I and CD4 down-regulation. Taken together, these results demonstrate that stress granule induction and nef mRNA sequestration account for this translational suppression of Nef expression and offers a new strategy for development of anti-HIV therapeutics to buttress our fight against HIV/AIDS. PMID:19150430

  6. Effects of HIV-1 Nef on retrograde transport from the plasma membrane to the endoplasmic reticulum.

    PubMed

    Johannes, Ludger; Pezo, Valérie; Mallard, Frédéric; Tenza, Danièle; Wiltz, Aimée; Saint-Pol, Agnès; Helft, Julie; Antony, Claude; Benaroch, Philippe

    2003-05-01

    HIV-1 Nef protein down-regulates several important immunoreceptors through interactions with components of the intracellular sorting machinery. Nef expression is also known to induce modifications of the endocytic pathway. Here, we analyzed the effects of Nef on retrograde transport, from the plasma membrane to the endoplasmic reticulum using Shiga toxin B-subunit (STxB). Nef expression inhibited access of STxB to the endoplasmic reticulum, but did not modify the surface expression level of STxB receptor, Gb3, nor its internalization rate as measured with a newly developed assay. Mutation of the myristoylation site or of a di-leucine motif of Nef involved in the interaction with the clathrin adaptor complexes AP1 and AP2 abolished the inhibition of retrograde transport. In contrast, mutations of Nef motifs known to interact with PACS-1, beta COP or a subunit of the v-ATPase did not modify the inhibitory activity of Nef on retrograde transport. Ultrastructural analysis revealed that Nef was present in clusters located on endosomal or Golgi membranes together with internalized STxB. Furthermore, in strongly Nef-expressing cells, STxB accumulated in endosomal structures that labeled with AP1. Our observations show that Nef perturbs retrograde transport between the early endosome and the endoplasmic reticulum. The potential transport steps targeted by Nef are discussed.

  7. HIV-1 Nef breaches placental barrier in rat model.

    PubMed

    Singh, Poonam; Agnihotri, Saurabh Kumar; Tewari, Mahesh Chandra; Kumar, Sadan; Sachdev, Monika; Tripathi, Raj Kamal

    2012-01-01

    The vertical transmission of HIV-1 from the mother to fetus is known, but the molecular mechanism regulating this transmission is not fully characterized. The fetus is highly protected by the placenta, which does not permit microbial pathogens to cross the placental barrier. In the present study, a rat model was established to observe the effect of HIV-1 protein Nef on placental barrier. Evans blue dye was used to assay permeability of placental barrier and fourteen day pregnant Sprague Dawley rats were injected intravenously with 2% Evans blue dye along with various concentrations of recombinant Nef. After an hour, animals were sacrificed and dye migration was observed through the assimilation of peripheral blood into fetus. Interestingly, traces of recombinant Nef protein were detected in the embryo as well as amniotic fluid and amniotic membrane along with placenta and uterus. Our study indicates that recombinant HIV-1-Nef protein breaches the placental barrier and allows the migration of Evans blue dye to the growing fetus. Further the concentration of Nef protein in blood is directly proportional to the intensity of dye migration and to the amount of Nef protein detected in uterus, placenta, amniotic membrane, amniotic fluid and embryo. Based on this study, it can be concluded that the HIV-1 Nef protein has a direct effect on breaching of the placental barrier in the model we have established in this study. Our observations will be helpful to understand the molecular mechanisms related to this breach of placental barrier by Nef in humans and may be helpful to identify specific Nef inhibitors.

  8. Is Nef the elusive cause of HIV-associated hematopoietic dysfunction?

    PubMed Central

    Kirchhoff, Frank; Silvestri, Guido

    2008-01-01

    HIV-associated hematological abnormalities involve all lineages of blood cells, thus implying that the virus impairs the function of early HSCs. However, the underlying mechanisms of this defect are unknown, particularly since HSCs are largely resistant to HIV-1 infection. In this issue of the JCI, Prost and colleagues show that the viral accessory protein Negative factor (Nef) plays a potentially critical role in the pathogenesis of HIV/SIV-associated hematopoietic dysfunction by affecting the clonogenic potential of HSCs (see the related article beginning on page 1765). Soluble Nef induces PPARγ in uninfected HSCs, thereby suppressing the expression of STAT5A and STAT5B, two factors necessary for proper HSC function. The identification of this novel activity of extracellular Nef defines a new mechanism of HIV/SIV pathogenesis and suggests that approaches aimed at increasing STAT5A and STAT5B expression may be considered in HIV-infected individuals with prominent hematological abnormalities. The results also raise the question of whether dysregulation of hematopoiesis by extracellular Nef plays a role in the development of T cell immunodeficiency and the high levels of chronic immune activation associated with AIDS. PMID:18431512

  9. Modulation of Different Human Immunodeficiency Virus Type 1 Nef Functions during Progression to AIDS

    PubMed Central

    Carl, Silke; Greenough, Thomas C.; Krumbiegel, Mandy; Greenberg, Michael; Skowronski, Jacek; Sullivan, John L.; Kirchhoff, Frank

    2001-01-01

    The human immunodeficiency virus type 1 (HIV-1) Nef protein has several independent functions that might contribute to efficient viral replication in vivo. Since HIV-1 adapts rapidly to its host environment, we investigated if different Nef properties are associated with disease progression. Functional analysis revealed that nef alleles obtained during late stages of infection did not efficiently downmodulate class I major histocompatibility complex but were highly active in the stimulation of viral replication. In comparison, functional activity in downregulation of CD4 and enhancement of HIV-1 infectivity were maintained or enhanced after AIDS progression. Our results demonstrate that various Nef activities are modulated during the course of HIV-1 infection to maintain high viral loads at different stages of disease progression. These findings suggest that all in vitro Nef functions investigated contribute to AIDS pathogenesis and indicate that nef variants with increased pathogenicity emerge in a significant number of HIV-1-infected individuals. PMID:11264355

  10. HIV-1 Nef mutations abrogating downregulation of CD4 affect other Nef functions and show reduced pathogenicity in transgenic mice.

    PubMed

    Hanna, Zaher; Priceputu, Elena; Hu, Chunyan; Vincent, Patrick; Jolicoeur, Paul

    2006-03-01

    HIV-1 Nef has the ability to downmodulate CD4 cell surface expression. Several studies have shown that CD4 downregulation is required for efficient virus replication and high infectivity. However, the pathophysiological relevance of this phenomenon in vivo, independently of its role in sustaining high virus loads, remains unclear. We studied the impact of the CD4 downregulation function of Nef on its pathogenesis in vivo, in the absence of viral replication, in the CD4C/HIV transgenic (Tg) mouse model. Two independent Nef mutants (RD35/36AA and D174K), known to abrogate CD4 downregulation, were tested in Tg mice. Flow cytometry analysis showed that downregulation of murine CD4 was severely decreased or abrogated on Tg T cells expressing respectively Nef(RD35/36AA) and Nef(D174K). Similarly, the severe depletion of double-positive CD4+CD8+ and of single-positive CD4+CD8- thymocytes, usually observed with Nef(Wt), was not detected in Nef(RD35/36AA) and Nef(D174K) Tg mice. However, both mutant Tg mice showed a partial depletion of peripheral CD4+ T cells. This was accompanied, as previously reported for Net(Wt) Tg mice, by the presence of an activated/memory-like phenotype (CD69+, CD25+, CD44+, CD45RB(Low), CD62(Low)) of CD4+ T cells expressing Nef(RD35/36AA) and to a lesser extent Nef(D174K). In addition, both mutants retained the ability to block CD4+ T cell proliferation in vitro after anti-CD3 stimulation, but not to enhance apoptosis/death of CD4+ T cells. Therefore, it appears that Nef-mediated CD4 downregulation is associated with thymic defects, but segregates independently of the activated/memory-like phenotype, of the partial depletion and of the impaired in vitro proliferation of peripheral CD4+ T cells. Histopathological assessment revealed the total absence of or decrease severity and frequency of organ AIDS-like diseases (lung, heart and kidney pathologies) in respectively Nef(RD35/36AA) and Nef(D174K) Tg mice, relative to those developing in Nef(Wt) Tg

  11. HIV-1 Nef mutations abrogating downregulation of CD4 affect other Nef functions and show reduced pathogenicity in transgenic mice

    SciTech Connect

    Hanna, Zaher . E-mail: Zaher.Hanna@ircm.qc.ca; Priceputu, Elena; Hu, Chunyan; Vincent, Patrick; Jolicoeur, Paul

    2006-03-01

    HIV-1 Nef has the ability to downmodulate CD4 cell surface expression. Several studies have shown that CD4 downregulation is required for efficient virus replication and high infectivity. However, the pathophysiological relevance of this phenomenon in vivo, independently of its role in sustaining high virus loads, remains unclear. We studied the impact of the CD4 downregulation function of Nef on its pathogenesis in vivo, in the absence of viral replication, in the CD4C/HIV transgenic (Tg) mouse model. Two independent Nef mutants (RD35/36AA and D174K), known to abrogate CD4 downregulation, were tested in Tg mice. Flow cytometry analysis showed that downregulation of murine CD4 was severely decreased or abrogated on Tg T cells expressing respectively Nef{sup RD35/36AA} and Nef{sup D174K}. Similarly, the severe depletion of double-positive CD4{sup +}CD8{sup +} and of single-positive CD4{sup +}CD8{sup -} thymocytes, usually observed with Nef{sup Wt}, was not detected in Nef{sup RD35/36AA} and Nef{sup D174K} Tg mice. However, both mutant Tg mice showed a partial depletion of peripheral CD4{sup +} T cells. This was accompanied, as previously reported for Net{sup Wt} Tg mice, by the presence of an activated/memory-like phenotype (CD69{sup +}, CD25{sup +}, CD44{sup +}, CD45RB{sup Low}, CD62{sup Low}) of CD4{sup +} T cells expressing Nef{sup RD35/36AA} and to a lesser extent Nef{sup D174K}. In addition, both mutants retained the ability to block CD4{sup +} T cell proliferation in vitro after anti-CD3 stimulation, but not to enhance apoptosis/death of CD4{sup +} T cells. Therefore, it appears that Nef-mediated CD4 downregulation is associated with thymic defects, but segregates independently of the activated/memory-like phenotype, of the partial depletion and of the impaired in vitro proliferation of peripheral CD4{sup +} T cells. Histopathological assessment revealed the total absence of or decrease severity and frequency of organ AIDS-like diseases (lung, heart and kidney

  12. HIV-1 Nef Induces CCL5 production in astrocytes through p38-MAPK and PI3K/Akt pathway and utilizes NF-kB, CEBP and AP-1 transcription factors

    NASA Astrophysics Data System (ADS)

    Liu, Xun; Shah, Ankit; Gangwani, Mohitkumar R.; Silverstein, Peter S.; Fu, Mingui; Kumar, Anil

    2014-03-01

    The prevalence of HIV-associated neurocognitive disorders (HAND) remains high in patients infected with HIV-1. The production of pro-inflammatory cytokines by astrocytes/microglia exposed to viral proteins is thought to be one of the mechanisms leading to HIV-1- mediated neurotoxicity. In the present study we examined the effects of Nef on CCL5 induction in astrocytes. The results demonstrate that CCL5 is significantly induced in Nef-transfected SVGA astrocytes. To determine the mechanisms responsible for the increased CCL5 caused by Nef, we employed siRNA and chemical antagonists. Antagonists of NF-κB, PI3K, and p38 significantly reduced the expression levels of CCL5 induced by Nef transfection. Furthermore, specific siRNAs demonstrated that the Akt, p38MAPK, NF-κB, CEBP, and AP-1 pathways play a role in Nef-mediated CCL5 expression. The results demonstrated that the PI3K/Akt and p38 MAPK pathways, along with the transcription factors NF-κB, CEBP, and AP-1, are involved in Nef-induced CCL5 production in astrocytes.

  13. Nef induces apoptosis by activating JNK signaling pathway and inhibits NF-kappaB-dependent immune responses in Drosophila.

    PubMed

    Lee, Sung Bae; Park, Jeehye; Jung, Jae U; Chung, Jongkyeong

    2005-05-01

    The human immunodeficiency virus type 1 (HIV-1) nef gene encodes a 27-kDa protein that plays a crucial role during AIDS pathogenesis, but its exact functional mechanism has not been fully elucidated and remains controversial. The present study illuminated the in vivo functions of Nef using Drosophila, in which genetic analyses can be conveniently conducted. Using Drosophila transgenic lines for wild-type Nef, we demonstrated that Nef is not involved in the regulation of cell proliferation but rather specifically induces caspase-dependent apoptosis in wings in a cell-autonomous manner. Interestingly, myristoylation-defective Nef completely failed to induce the apoptotic wing phenotypes, consistent with previous reports demonstrating a crucial role for membrane localization of Nef in vivo. Further genetic and immunohistochemical studies revealed that Nef-dependent JNK activation is responsible for apoptosis. Furthermore, we found that ectopic expression of Nef inhibits Drosophila innate immune responses including Relish NF-kappaB activation with subsequent induction of an antimicrobial peptide, diptericin. The in vivo functions of Nef in Drosophila are highly consistent with those found in mammals and so we propose that Nef regulates evolutionarily highly conserved signaling molecules of the JNK and NF-kappaB signaling pathways at the plasma membrane, and consequently modulates apoptosis and immune responses in HIV target cells.

  14. Refined solution structure and backbone dynamics of HIV-1 Nef.

    PubMed Central

    Grzesiek, S.; Bax, A.; Hu, J. S.; Kaufman, J.; Palmer, I.; Stahl, S. J.; Tjandra, N.; Wingfield, P. T.

    1997-01-01

    The tendency of HIV-1 Nef to form aggregates in solution, particularly at pH values below 8, together with its large fraction of highly mobile residues seriously complicated determination of its three-dimensional structure, both for heteronuclear solution NMR (Grzesiek et al., 1996a, Nat Struct Biol 3:340-345) and for X-ray crystallography (Lee et al., 1996, Cell 85:931-942). Methods used to determine the Nef structure by NMR at pH 8 and 0.6 mM concentration are presented, together with a detailed description of Nef's secondary and tertiary structure. The described techniques have general applicability for the NMR structure determination of proteins that are aggregating and/or have limited stability at low pH values. Extensive chemical shift assignments are reported for backbone and side chain 1H, 13C, and 15N resonances of the HIV-1 Nef deletion mutants NEF delta 2-39, NEF delta 2-39, delta 159-173, and of NEF delta 2-39, delta 159-173 in complex with the SH3 domain of the Hck tyrosine protein kinase. Besides a type II polyproline helix, Nef's structure consists of three alpha-helices, a 3(10) helix, and a five-stranded anti-parallel beta-sheet. The analysis of 15N relaxation parameters of the backbone amide sites reveals that all the secondary structure elements are non-mobile on the picosecond to nanosecond and on the millisecond time scale. A large number of slowly exchanging amide protons provides evidence for the stability of the Nef core even on the time scale of hours. Significant internal motions on the ps to ns time scale are detected for residues 60 to 71 and for residues 149 to 180, which form solvent-exposed loops. The residues of the HIV-1 protease cleavage site (W57/L58) do not exhibit large amplitude motions on the sub-nanosecond time scale, and their side chains insert themselves into a hydrophobic crevice formed between the C-terminus of helix 1 and the N-terminus of helix 2. A refined structure has been determined based on additional constraints

  15. Genetic and Functional Diversity of Human Immunodeficiency Virus Type 1 Subtype B Nef Primary Isolates

    PubMed Central

    Foster, John L.; Molina, Rene P.; Luo, Tianci; Arora, Vivek K.; Huang, Yaoxing; Ho, David D.; Garcia, J. Victor

    2001-01-01

    We have characterized the functional integrity of seven primary Nef isolates: five from a long-term nonprogressing human immunodeficiency virus (HIV)-infected individual and one each from two patients with AIDS. One of the seven Nefs was defective for CD4 downregulation, two others were defective for PAK-2 activation, and one Nef was defective for PAK-2 activation and major histocompatibility complex (MHC) class I downregulation. Five of the Nefs were tested and found to be functional for the enhancement of virus particle infectivity. The structural basis for each of the functional defects has been analyzed by constructing a consensus nef, followed by mutational analysis of the variant amino acid residues. Mutations A29V and F193I were deleterious to CD4 downregulation and PAK-2 activation, respectively, while S189R rendered Nef defective for both MHC class I downregulation and PAK-2 activation. A search of the literature identified HIVs from five patients with Nefs predominantly mutated at F193 and from one patient with Nefs predominantly mutated at A29. A29 is highly conserved in all HIV subtypes except for subtype E. F193 is conserved in subtype B (and possibly in the closely related subtype D), but none of the other HIV group M subtypes. Our results suggest that functional distinctions may exist between HIV subtypes. PMID:11160665

  16. Overlapping effector interfaces define the multiple functions of the HIV-1 Nef polyproline helix

    PubMed Central

    2012-01-01

    C- terminal sides of the polyproline helix are multifunctional protein interaction sites. The polyproline segment is also adapted to downregulate MHCI with a non-canonical binding surface. Our results demonstrate that Nef polyproline helix is highly adapted to directly interact with multiple host cell proteins. PMID:22651890

  17. Overlapping effector interfaces define the multiple functions of the HIV-1 Nef polyproline helix.

    PubMed

    Kuo, Lillian S; Baugh, Laura L; Denial, Sarah J; Watkins, Richard L; Liu, Mingjie; Garcia, J Victor; Foster, John L

    2012-05-31

    helix are multifunctional protein interaction sites. The polyproline segment is also adapted to downregulate MHCI with a non-canonical binding surface. Our results demonstrate that Nef polyproline helix is highly adapted to directly interact with multiple host cell proteins.

  18. HIV-1 Nef regulates the release of superoxide anions from human macrophages

    PubMed Central

    2005-01-01

    The NADPH oxidase enzymatic complex participates in the oxidative burst by producing ROS (reactive oxygen species). Altered levels of ROS production may have pathogenetic implications due to the loss of some innate immune functions such as oxidative burst and phagocytosis. Considering that HIV-1 Nef protein plays a primary role in AIDS pathogenesis, by affecting the immune system, we sought to dissect possible effects of Nef on the release of superoxide anions. We show here that the inducible expression of Nef in human phagocytic cells modulates the superoxide release in a biphasic manner. In particular, an early Nef-induced increase of the superoxide release was followed by a dramatic decrease starting from 10 h after the Nef induction. This was observed whatever the presence of cell activators such as GM-CSF (granulocyte/macrophage colony-stimulating factor) or fMLP (N-formyl-L-methionyl-L-leucyl-L-phenylalanine). Whereas the early increase in superoxide release is probably the result of the already described Nef-dependent activation of PAK-2 (p21-activated kinase 2)–Rac2, we were interested in investigating the mechanisms underlying the late inhibition of superoxide release observed originally. In this regard, we individuated at least three independent requirements for the Nef-induced blockade of superoxide release: (i) the active protein synthesis; (ii) both the membrane localization and the interaction with endocytotic machinery of Nef; and (iii) the release of soluble factor(s). Moreover, we observed that IL-10 (interleukin-10) inhibits superoxide release, whereas its depletion restored NADPH oxidase activity. We propose that the cell membrane-to-lysosome Nef transit leads to the synthesis and release of soluble factor(s) and, among them, IL-10 might significantly contribute to the inhibition of NAPDH oxidase activity. PMID:15847608

  19. Lentiviral Nef proteins manipulate T cells in a subset-specific manner.

    PubMed

    Yu, Hangxing; Khalid, Mohammad; Heigele, Anke; Schmökel, Jan; Usmani, Shariq M; van der Merwe, Johannes; Münch, Jan; Silvestri, Guido; Kirchhoff, Frank

    2015-02-01

    The role of the accessory viral Nef protein as a multifunctional manipulator of the host cell that is required for effective replication of human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) in vivo is well established. It is unknown, however, whether Nef manipulates all or just specific subsets of CD4(+) T cells, which are the main targets of virus infection and differ substantially in their state of activation and importance for a functional immune system. Here, we analyzed the effect of Nef proteins differing in their T cell receptor (TCR)-CD3 downmodulation function in HIV-infected human lymphoid aggregate cultures and peripheral blood mononuclear cells. We found that Nef efficiently downmodulates TCR-CD3 in naive and memory CD4(+) T cells and protects the latter against apoptosis. In contrast, highly proliferative CD45RA(+) CD45RO(+) CD4(+) T cells were main producers of infectious virus but largely refractory to TCR-CD3 downmodulation. Such T cell subset-specific differences were also observed for Nef-mediated modulation of CD4 but not for enhancement of virion infectivity. Our results indicate that Nef predominantly modulates surface receptors on CD4(+) T cell subsets that are not already fully permissive for viral replication. As a consequence, Nef-mediated downmodulation of TCR-CD3, which distinguishes most primate lentiviruses from HIV type 1 (HIV-1) and its vpu-containing simian precursors, may promote a selective preservation of central memory CD4(+) T cells, which are critical for the maintenance of a functional immune system. The Nef proteins of human and simian immunodeficiency viruses manipulate infected CD4(+) T cells in multiple ways to promote viral replication and immune evasion in vivo. Here, we show that some effects of Nef are subset specific. Downmodulation of CD4 and TCR-CD3 is highly effective in central memory CD4(+) T cells, and the latter Nef function protects this T cell subset against apoptosis. In contrast

  20. Lentiviral Nef Proteins Manipulate T Cells in a Subset-Specific Manner

    PubMed Central

    Yu, Hangxing; Khalid, Mohammad; Heigele, Anke; Schmökel, Jan; M. Usmani, Shariq; van der Merwe, Johannes; Münch, Jan; Silvestri, Guido

    2014-01-01

    ABSTRACT The role of the accessory viral Nef protein as a multifunctional manipulator of the host cell that is required for effective replication of human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) in vivo is well established. It is unknown, however, whether Nef manipulates all or just specific subsets of CD4+ T cells, which are the main targets of virus infection and differ substantially in their state of activation and importance for a functional immune system. Here, we analyzed the effect of Nef proteins differing in their T cell receptor (TCR)-CD3 downmodulation function in HIV-infected human lymphoid aggregate cultures and peripheral blood mononuclear cells. We found that Nef efficiently downmodulates TCR-CD3 in naive and memory CD4+ T cells and protects the latter against apoptosis. In contrast, highly proliferative CD45RA+ CD45RO+ CD4+ T cells were main producers of infectious virus but largely refractory to TCR-CD3 downmodulation. Such T cell subset-specific differences were also observed for Nef-mediated modulation of CD4 but not for enhancement of virion infectivity. Our results indicate that Nef predominantly modulates surface receptors on CD4+ T cell subsets that are not already fully permissive for viral replication. As a consequence, Nef-mediated downmodulation of TCR-CD3, which distinguishes most primate lentiviruses from HIV type 1 (HIV-1) and its vpu-containing simian precursors, may promote a selective preservation of central memory CD4+ T cells, which are critical for the maintenance of a functional immune system. IMPORTANCE The Nef proteins of human and simian immunodeficiency viruses manipulate infected CD4+ T cells in multiple ways to promote viral replication and immune evasion in vivo. Here, we show that some effects of Nef are subset specific. Downmodulation of CD4 and TCR-CD3 is highly effective in central memory CD4+ T cells, and the latter Nef function protects this T cell subset against apoptosis. In contrast

  1. Synthesis and Structure-activity Analysis of Diphenylpyrazolodiazene Inhibitors of the HIV-1 Nef Virulence Factor

    PubMed Central

    Iyer, Prema C.; Zhao, Jielu; Emert-Sedlak, Lori A.; Moore, Kerry; Smithgall, Thomas E.; Day, Billy W.

    2014-01-01

    HIV-1 Nef is a critical AIDS progression factor yet underexplored target for antiretroviral drug discovery. A recent high-throughput screen for pharmacological inhibitors of Nef-dependent Src-family kinase activation identified a diphenylpyrazolodiazene hit compound with submicromolar potency in HIV-1 replication assays against a broad range of primary Nef variants. This compound, known as ‘B9’, binds directly to Nef and inhibits is dimerization in cells as a possible mechanism of action. Here were synthesized a diverse set of B9 analogs and identified structural features essential to antiretroviral activity. Chemical modifications to each of the three rings present in the parent compound were identified that did not compromise antiviral action. These analogs will guide the development of next-generation compounds with appropriate pharmacological profiles for assessment of antiretroviral activity in vivo. PMID:24650642

  2. The interaction between HIV-1 Nef and adaptor protein-2 reduces Nef-mediated CD4(+) T cell apoptosis.

    PubMed

    Jacob, Rajesh Abraham; Johnson, Aaron L; Pawlak, Emily N; Dirk, Brennan S; Van Nynatten, Logan R; Haeryfar, S M Mansour; Dikeakos, Jimmy D

    2017-09-01

    Acquired Immune Deficiency Syndrome is characterized by a decline in CD4(+) T cells. Here, we elucidated the mechanism underlying apoptosis in Human Immunodeficiency Virus-1 (HIV-1) infection by examining host apoptotic pathways hijacked by the HIV-1 Nef protein in the CD4(+) T-cell line Sup-T1. Using a panel of Nef mutants unable to bind specific host proteins we uncovered that Nef generates pro- and anti-apoptotic signals. Apoptosis increased upon mutating the motifs involved in the interaction of Nef:AP-1 (NefM20A or NefEEEE62-65AAAA) or Nef:AP-2 (NefLL164/165AA), implying these interactions limit Nef-mediated apoptosis. In contrast, disrupting the Nef:PAK2 interaction motifs (NefH89A or NefF191A) reduced apoptosis. To validate further, apoptosis was measured after short-hairpin RNA knock-down of AP-1, AP-2 and PAK2. AP-2α depletion enhanced apoptosis, demonstrating that disrupting the Nef:AP-2α interaction limits Nef-mediated apoptosis. Collectively, we describe a mechanism by which HIV-1 regulates cell survival and demonstrate the consequence of interfering with Nef:host protein interactions. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Functional adaptation of Nef to the immune milieu of HIV-1 infection in vivo.

    PubMed

    Lewis, Martha J; Balamurugan, Arumugam; Ohno, Ayako; Kilpatrick, Stephanie; Ng, Hwee L; Yang, Otto O

    2008-03-15

    Nef-mediated down-regulation of MHC class I (MHC-I) molecules on HIV-1-infected cells has been proposed to enhance viral persistence through evasion of host CTLs. This conclusion is based largely on demonstrations that Nef from laboratory HIV-1 strains reduces the susceptibility of infected cells to CTL killing in vitro. However, the function and role of Nef-mediated MHC-I down-regulation in vivo have not been well described. To approach this issue, nef quasispecies from chronically HIV-1-infected individuals were cloned into recombinant reporter viruses and tested for their ability to down-regulate MHC-I molecules from the surface of infected cells. The level of function varied widely between individuals, and although comparison to the immunologic parameters of blood CD4(+) T lymphocyte count and breadth of the HIV-1-specific CTL response showed positive correlations, no significant correlation was found in comparison to plasma viremia. The ability of in vivo-derived Nef to down-regulate MHC-I predicted the resistance of HIV-1 to suppression by CTL. Taken together, these data demonstrate the functionality of Nef to down-regulate MHC-I in vivo during stable chronic infection, and suggest that this function is maintained by the need of HIV-1 to cope with the antiviral CTL response.

  4. CD4 and MHC class I down-modulation activities of nef alleles from brain- and lymphoid tissue-derived primary HIV-1 isolates

    PubMed Central

    Gray, Lachlan R.; Gabuzda, Dana; Cowley, Daniel; Ellett, Anne; Chiavaroli, Lisa; Wesselingh, Steven L.; Churchill, Melissa J.; Gorry, Paul R.

    2015-01-01

    HIV-1 nef undergoes adaptive evolution in the CNS, reflecting altered requirements for HIV-1 replication in macrophages/microglia and brain-specific immune selection pressures. The role of Nef in HIV-1 neurotropism and the pathogenesis of HIV-associated dementia (HAD) is unclear. In this study, we characterized 82 nef alleles cloned from brain, CSF, spinal cord and blood/lymphoid tissue-derived HIV-1 isolates from 7 subjects with HAD. CNS isolate-derived nef alleles were genetically compartmentalized and had reduced sequence diversity compared to those from lymphoid tissue isolates. Defective nef alleles predominated in a brain-derived isolate from one of the 7 subjects (MACS2-br). The ability of Nef to down-modulate CD4 and MHC class 1 (MHC-1) was generally conserved among nef alleles from both CNS and lymphoid tissues. However, the potency of CD4 and MHC-1 down-modulation was variable, which was associated with sequence alterations known to influence these Nef functions. These results suggest that CD4 and MHC-1 down-modulation are highly conserved functions among nef alleles from CNS- and lymphoid tissue-derived HIV-1 isolates that may contribute to viral replication and escape from immune surveillance in the CNS. PMID:21165790

  5. CD4 cell surface downregulation in HIV-1 Nef transgenic mice is a consequence of intracellular sequestration.

    PubMed Central

    Brady, H J; Pennington, D J; Miles, C G; Dzierzak, E A

    1993-01-01

    The Nef gene product is a regulatory protein of HIV whose biological function is poorly understood. Nef has been thought to have a negative effect on viral replication in vitro but has been shown in studies with SIV to be necessary in the establishment of viraemia in vivo. In vitro studies in various human cell lines have shown that Nef downregulates the expression of cell surface CD4 and thus could have effects on the immune response. We have generated four transgenic mouse lines, with constructs containing two different Nef alleles under the control of CD2 regulatory elements to examine the interaction of Nef with the host immune system in vivo. In adult transgenic mice we have found marked downregulation in the level of CD4 on the surface of double positive thymocytes and a decrease in the number of CD4+ T cells in the thymus. Functional analyses have revealed a decrease in the total activation of transgenic thymocytes by anti-CD3 epsilon antibody. By specific intracellular staining of T cells in such mice we have found CD4 colocalizing with a Golgi-specific marker. These results strongly suggest a Nef mediated effect on developing CD4 thymocytes resulting from interference of Nef in the intracellular trafficking or post-translational modification of CD4. Images PMID:8262036

  6. Changes in the Plasticity of HIV-1 Nef RNA during the Evolution of the North American Epidemic

    PubMed Central

    Manzourolajdad, Amirhossein; Gonzalez, Mileidy; Spouge, John L.

    2016-01-01

    Because of a high mutation rate, HIV exists as a viral swarm of many sequence variants evolving under various selective pressures from the human immune system. Although the Nef gene codes for the most immunogenic of HIV accessory proteins, which alone makes it of great interest to HIV research, it also encodes an RNA structure, whose contribution to HIV virulence has been largely unexplored. Nef RNA helps HIV escape RNA interference (RNAi) through nucleotide changes and alternative folding. This study examines Historic and Modern Datasets of patient HIV-1 Nef sequences during the evolution of the North American epidemic for local changes in RNA plasticity. By definition, RNA plasticity refers to an RNA molecule’s ability to take alternative folds (i.e., alternative conformations). Our most important finding is that an evolutionarily conserved region of the HIV-1 Nef gene, which we denote by R2, recently underwent a statistically significant increase in its RNA plasticity. Thus, our results indicate that Modern Nef R2 typically accommodates an alternative fold more readily than Historic Nef R2. Moreover, the increase in RNA plasticity resides mostly in synonymous nucleotide changes, which cannot be a response to selective pressures on the Nef protein. R2 may therefore be of interest in the development of antiviral RNAi therapies. PMID:27685447

  7. HIV-1 Nef responsiveness is determined by Env variable regions involved in trimer association and correlates with neutralization sensitivity.

    PubMed

    Usami, Yoshiko; Göttlinger, Heinrich

    2013-11-14

    HIV-1 Nef and the unrelated murine leukemia virus glycoGag similarly enhance the infectivity of HIV-1 virions. We now show that the effects of Nef and glycoGag are similarly determined by variable regions of HIV-1 gp120 that control Env trimer association and neutralization sensitivity. Whereas neutralization-sensitive X4-tropic Env proteins conferred high responsiveness to Nef and glycoGag, particles bearing neutralization-resistant R5-tropic Envs were considerably less affected. The profoundly different Nef/glycoGag responsiveness of a neutralization-resistant and a neutralization-sensitive R5-tropic Env could be switched by exchanging their gp120 V1/V2 regions, which also switches their neutralization sensitivity. Within V1/V2, the same determinants governed Nef/glycoGag responsiveness and neutralization sensitivity, indicating that these phenotypes are mechanistically linked. The V1/V2 and V3 regions, which form an apical trimer-association domain, together determined the Nef and glycoGag responsiveness of an X4-tropic Env. Our results suggest that Nef and glycoGag counteract the inactivation of Env spikes with relatively unstable apical trimer-association domains.

  8. HIV Nef expression favors the relative preservation of CD4+ T regulatory cells that retain some important suppressive functions.

    PubMed

    Chrobak, Pavel; Afkhami, Soheila; Priceputu, Elena; Poudrier, Johanne; Meunier, Clémence; Hanna, Zaher; Sparwasser, Tim; Jolicoeur, Paul

    2014-02-15

    HIV-1 infection causes depletion and/or dysfunction of distinct CD4(+) T cell subsets and may affect these differently. Using the CD4C/HIV-1(Nef) transgenic (Tg) mice as a model, we report that HIV-1 Nef causes depletion of total CD4(+) T cells, but preserves and relatively enriches CD4(+) regulatory T cells (Treg). We found that Nef-mediated CD4(+) Treg enrichment is the direct result of Nef expression in CD4(+) T cells, occurs independently of Nef-induced lymphopenia, and most likely results from multiple mechanisms: lower apoptosis, enhanced cell division, and increased generation from precursors. Interestingly, Tg Treg relative enrichment could be reversed by enhancing Lck activity. Most importantly, we show that, in contrast to Tg helper CD4(+) T cells that have lost their function, Nef-expressing CD4(+) Treg retain their regulatory function in vitro and also in vivo, under some settings. In particular, we found that Treg prevent expansion of Tg B and non-Treg T cells in vivo. Our study reveals that Nef affects distinct CD4(+) T cell subsets differently and uncovers the high proliferative potential of B and non-Treg T cells in this mouse model.

  9. Impaired Nef function is associated with early control of HIV-1 viremia.

    PubMed

    Kuang, Xiaomei T; Li, Xiaoguang; Anmole, Gursev; Mwimanzi, Philip; Shahid, Aniqa; Le, Anh Q; Chong, Louise; Qian, Hua; Miura, Toshiyuki; Markle, Tristan; Baraki, Bemuluyigza; Connick, Elizabeth; Daar, Eric S; Jessen, Heiko; Kelleher, Anthony D; Little, Susan; Markowitz, Martin; Pereyra, Florencia; Rosenberg, Eric S; Walker, Bruce D; Ueno, Takamasa; Brumme, Zabrina L; Brockman, Mark A

    2014-09-01

    Host and viral factors influence the HIV-1 infection course. Reduced Nef function has been observed in HIV-1 controllers during the chronic phase, but the kinetics and mechanisms of Nef attenuation in such individuals remain unclear. We examined plasma RNA-derived Nef clones from 10 recently infected individuals who subsequently suppressed viremia to less than 2,000 RNA copies/ml within 1 year postinfection (acute controllers) and 50 recently infected individuals who did not control viremia (acute progressors). Nef clones from acute controllers displayed a lesser ability to downregulate CD4 and HLA class I from the cell surface and a reduced ability to enhance virion infectivity compared to those from acute progressors (all P<0.01). HLA class I downregulation activity correlated inversely with days postinfection (Spearman's R=-0.85, P=0.004) and positively with baseline plasma viral load (Spearman's R=0.81, P=0.007) in acute controllers but not in acute progressors. Nef polymorphisms associated with functional changes over time were identified in follow-up samples from six controllers. For one such individual, mutational analyses indicated that four polymorphisms selected by HLA-A*31 and B*37 acted in combination to reduce Nef steady-state protein levels and HLA class I downregulation activity. Our results demonstrate that relative control of initial HIV-1 viremia is associated with Nef clones that display reduced function, which in turn may influence the course of HIV-1 infection. Transmission of impaired Nef sequences likely contributed in part to this observation; however, accumulation of HLA-associated polymorphisms in Nef that impair function also suggests that CD8+ T-cell pressures play a role in this phenomenon. Rare individuals can spontaneously control HIV-1 viremia in the absence of antiretroviral treatment. Understanding the host and viral factors that contribute to the controller phenotype may identify new strategies to design effective vaccines or

  10. Impaired Nef Function Is Associated with Early Control of HIV-1 Viremia

    PubMed Central

    Kuang, Xiaomei T.; Li, Xiaoguang; Anmole, Gursev; Mwimanzi, Philip; Shahid, Aniqa; Le, Anh Q.; Chong, Louise; Qian, Hua; Miura, Toshiyuki; Markle, Tristan; Baraki, Bemuluyigza; Connick, Elizabeth; Daar, Eric S.; Jessen, Heiko; Kelleher, Anthony D.; Little, Susan; Markowitz, Martin; Pereyra, Florencia; Rosenberg, Eric S.; Walker, Bruce D.; Ueno, Takamasa; Brumme, Zabrina L.

    2014-01-01

    ABSTRACT Host and viral factors influence the HIV-1 infection course. Reduced Nef function has been observed in HIV-1 controllers during the chronic phase, but the kinetics and mechanisms of Nef attenuation in such individuals remain unclear. We examined plasma RNA-derived Nef clones from 10 recently infected individuals who subsequently suppressed viremia to less than 2,000 RNA copies/ml within 1 year postinfection (acute controllers) and 50 recently infected individuals who did not control viremia (acute progressors). Nef clones from acute controllers displayed a lesser ability to downregulate CD4 and HLA class I from the cell surface and a reduced ability to enhance virion infectivity compared to those from acute progressors (all P < 0.01). HLA class I downregulation activity correlated inversely with days postinfection (Spearman's R = −0.85, P = 0.004) and positively with baseline plasma viral load (Spearman's R = 0.81, P = 0.007) in acute controllers but not in acute progressors. Nef polymorphisms associated with functional changes over time were identified in follow-up samples from six controllers. For one such individual, mutational analyses indicated that four polymorphisms selected by HLA-A*31 and B*37 acted in combination to reduce Nef steady-state protein levels and HLA class I downregulation activity. Our results demonstrate that relative control of initial HIV-1 viremia is associated with Nef clones that display reduced function, which in turn may influence the course of HIV-1 infection. Transmission of impaired Nef sequences likely contributed in part to this observation; however, accumulation of HLA-associated polymorphisms in Nef that impair function also suggests that CD8+ T-cell pressures play a role in this phenomenon. IMPORTANCE Rare individuals can spontaneously control HIV-1 viremia in the absence of antiretroviral treatment. Understanding the host and viral factors that contribute to the controller phenotype may identify new strategies to

  11. Nef Neutralizes the Ability of Exosomes from CD4+ T Cells to Act as Decoys during HIV-1 Infection

    PubMed Central

    da Silva, Elaine Z. M.; Silveira, Paola P.; da Silva-Januário, Mara E.; Arruda, Eurico; Jamur, Maria C.; Oliver, Constance; Aguiar, Renato S.; daSilva, Luis L. P.

    2014-01-01

    Nef is an HIV-1 accessory protein that promotes viral replication and pathogenesis. A key function of Nef is to ensure sustained depletion of CD4 and MHC-I molecules in infected cells by inducing targeting of these proteins to multivesicular bodies (MVBs), and ultimately to lysosomes for degradation. Nef also affects cellular secretory routes promoting its own secretion via exosomes. To better understand the effects of Nef on the exocytic pathway, we investigated whether this viral factor modifies the composition of exosomes released by T lymphocytes. We showed that both CD4 and MHC-I molecules are secreted in exosomes from T cells and that the expression of Nef reduces the amount of these proteins in exosomes. To investigate the functional role for this novel activity of Nef, we performed in vitro HIV-1 infection assays in the presence of distinct populations of exosomes. We demonstrated that exosomes released by CD4+ T cells, but not CD4− T cells, efficiently inhibit HIV-1 infection in vitro. Because CD4 is the main receptor for HIV-1 infection, these results suggest that CD4 molecules displayed on the surface of exosomes can bind to envelope proteins of HIV-1 hindering virus interaction with target cells and infection. Importantly, CD4-depleted exosomes released by CD4+ T cells expressing Nef have a reduced capacity to inhibit HIV-1 infection in vitro. These results provide evidence that Nef promotes HIV-1 infection by reducing the expression of CD4 in exosomes from infected cells, besides the original role of Nef in reducing the CD4 levels at the cell surface. PMID:25423108

  12. Reduced cell surface expression of processed human immunodeficiency virus type 1 envelope glycoprotein in the presence of Nef.

    PubMed Central

    Schwartz, O; Rivière, Y; Heard, J M; Danos, O

    1993-01-01

    nef genes from two laboratory grown human immunodeficiency virus type 1 (HIV-1) strains and from two proviruses that had not been propagated in vitro were introduced into CD4+ lymphoblastoid CEM cells. The stable expression of all four Nef proteins was associated with an almost complete abrogation of CD4 cell surface localization. The consequences of the presence of Nef on gp160 cleavage, gp120 surface localization, and envelope-induced cytopathic effect were examined in CEM cells in which the HIV-1 env gene was expressed from a vaccinia virus vector. The presence of Nef did not modify the processing of gp160 into its subunits but resulted in a significant decrease of cell surface levels of gp120, associated with a dramatic reduction of the fusion-mediated cell death. Surface levels of mutant envelope glycoproteins unable to bind CD4 were not altered in Nef-expressing cells, suggesting that the phenomenon was CD4 dependent. The intracellular accumulation of fully processed envelope glycoproteins could significantly delay the cytopathic effect associated with envelope surface expression in HIV-infected cells and may be relevant to the selective advantage associated with Nef during the in vivo infectious process. Images PMID:8497051

  13. Reacquisition of Nef-mediated tetherin antagonism in a single in vivo passage of HIV-1 through its original chimpanzee host

    PubMed Central

    Götz, Nicola; Sauter, Daniel; Usmani, Shariq M.; Fritz, Joëlle V.; Goffinet, Christine; Heigele, Anke; Geyer, Matthias; Bibollet-Ruche, Frederic; Learn, Gerald H.; Fackler, Oliver T.; Hahn, Beatrice H.; Kirchhoff, Frank

    2012-01-01

    SUMMARY The interferon-induced host restriction factor tetherin poses a barrier for SIV transmission from primates to humans. Following cross-species transmission, the chimpanzee precursor of pandemic HIV-1 switched from the accessory protein Nef to Vpu to effectively counteract human tetherin. As we report here, the experimental reintroduction of HIV-1 into its original chimpanzee host resulted in a virus that can use both Vpu and Nef to antagonize chimpanzee tetherin. Functional analyses demonstrated that alterations in and near the highly conserved ExxxLL motif in the C-terminal loop of Nef were critical for the reacquisition of anti-tetherin activity. Strikingly, just two amino acid changes allowed HIV-1 Nef to counteract chimpanzee tetherin and promote virus release. Our data demonstrate that primate lentiviruses can reacquire lost accessory gene functions during a single in vivo passage and suggest that other functional constraints keep Nef ready to regain anti-tetherin activity. PMID:22980333

  14. Consequences of HLA-B*13-Associated Escape Mutations on HIV-1 Replication and Nef Function

    PubMed Central

    Shahid, Aniqa; Olvera, Alex; Anmole, Gursev; Kuang, Xiaomei T.; Cotton, Laura A.; Plana, Montserrat; Brander, Christian; Brockman, Mark A.

    2015-01-01

    ABSTRACT HLA-B*13 is associated with superior in vivo HIV-1 viremia control. Protection is thought to be mediated by sustained targeting of key cytotoxic T lymphocyte (CTL) epitopes and viral fitness costs of CTL escape in Gag although additional factors may contribute. We assessed the impact of 10 published B*13-associated polymorphisms in Gag, Pol, and Nef, in 23 biologically relevant combinations, on HIV-1 replication capacity and Nef-mediated reduction of cell surface CD4 and HLA class I expression. Mutations were engineered into HIV-1NL4.3, and replication capacity was measured using a green fluorescent protein (GFP) reporter T cell line. Nef-mediated CD4 and HLA-A*02 downregulation was assessed by flow cytometry, and T cell recognition of infected target cells was measured via coculture with an HIV-specific luciferase reporter cell line. When tested individually, only Gag-I147L and Gag-I437L incurred replicative costs (5% and 17%, respectively), consistent with prior reports. The Gag-I437L-mediated replication defect was rescued to wild-type levels by the adjacent K436R mutation. A novel B*13 epitope, comprising 8 residues and terminating at Gag147, was identified in p24Gag (GQMVHQAIGag140–147). No other single or combination Gag, Pol, or Nef mutant impaired viral replication. Single Nef mutations did not affect CD4 or HLA downregulation; however, the Nef double mutant E24Q-Q107R showed 40% impairment in HLA downregulation with no evidence of Nef stability defects. Moreover, target cells infected with HIV-1-NefE24Q-Q107R were recognized better by HIV-specific T cells than those infected with HIV-1NL4.3 or single Nef mutants. Our results indicate that CTL escape in Gag and Nef can be functionally costly and suggest that these effects may contribute to long-term HIV-1 control by HLA-B*13. IMPORTANCE Protective effects of HLA-B*13 on HIV-1 disease progression are mediated in part by fitness costs of CTL escape mutations in conserved Gag epitopes, but other

  15. Consequences of HLA-B*13-Associated Escape Mutations on HIV-1 Replication and Nef Function.

    PubMed

    Shahid, Aniqa; Olvera, Alex; Anmole, Gursev; Kuang, Xiaomei T; Cotton, Laura A; Plana, Montserrat; Brander, Christian; Brockman, Mark A; Brumme, Zabrina L

    2015-11-01

    HLA-B*13 is associated with superior in vivo HIV-1 viremia control. Protection is thought to be mediated by sustained targeting of key cytotoxic T lymphocyte (CTL) epitopes and viral fitness costs of CTL escape in Gag although additional factors may contribute. We assessed the impact of 10 published B*13-associated polymorphisms in Gag, Pol, and Nef, in 23 biologically relevant combinations, on HIV-1 replication capacity and Nef-mediated reduction of cell surface CD4 and HLA class I expression. Mutations were engineered into HIV-1NL4.3, and replication capacity was measured using a green fluorescent protein (GFP) reporter T cell line. Nef-mediated CD4 and HLA-A*02 downregulation was assessed by flow cytometry, and T cell recognition of infected target cells was measured via coculture with an HIV-specific luciferase reporter cell line. When tested individually, only Gag-I147L and Gag-I437L incurred replicative costs (5% and 17%, respectively), consistent with prior reports. The Gag-I437L-mediated replication defect was rescued to wild-type levels by the adjacent K436R mutation. A novel B*13 epitope, comprising 8 residues and terminating at Gag147, was identified in p24(Gag) (GQMVHQAIGag140-147). No other single or combination Gag, Pol, or Nef mutant impaired viral replication. Single Nef mutations did not affect CD4 or HLA downregulation; however, the Nef double mutant E24Q-Q107R showed 40% impairment in HLA downregulation with no evidence of Nef stability defects. Moreover, target cells infected with HIV-1-NefE24Q-Q107R were recognized better by HIV-specific T cells than those infected with HIV-1NL4.3 or single Nef mutants. Our results indicate that CTL escape in Gag and Nef can be functionally costly and suggest that these effects may contribute to long-term HIV-1 control by HLA-B*13. Protective effects of HLA-B*13 on HIV-1 disease progression are mediated in part by fitness costs of CTL escape mutations in conserved Gag epitopes, but other mechanisms remain

  16. The membrane-associated conformation of HIV-1 Nef investigated with hydrogen exchange mass spectrometry at a Langmuir monolayer

    PubMed Central

    Pirrone, Gregory F.; Emert-Sedlak, Lori A.; Wales, Thomas E.; Smithgall, Thomas E.; Kent, Michael S.; Engen, John R.

    2015-01-01

    In the companion paper to this work, we have described development of a new type of hydrogen exchange (HX) mass spectrometry (MS) measurement that integrates Langmuir monolayers. With Langmuir monolayers, the lipid packing density can be reproducibly controlled and changed as desired. Analysis of HX in proteins that may undergo conformational changes as a function of lipid packing, for example conformational rearrangements after insertion into a lipid layer, are then possible. We previously used neutron reflection to characterize just such a conformational change in the myristoylated HIV-1 Nef protein (myrNef): at high lipid packing density, myrNef could not insert into the lipids and maintained a compact conformation adjacent to the monolayer whereas at lower lipid packing density, myrNef was able to insert N-terminal arm residues causing displacement of the core domain away from the monolayer. In order to locate where conformation may have been altered by lipid association, we applied the HX MS Langmuir monolayer method to myrNef associated with monolayers of packing densities identical to those used for the prior neutron reflection measurements. The results show that the N-terminal region and the C-terminal unstructured loop undergo conformational changes when associated with a low lipid density lipid monolayer. The results are not consistent with the hypothesis of myrNef dimerization upon membrane association in the absence of other myrNef binding partners. The HX MS Langmuir monolayer method provides new and meaningful information for myrNef that helps explain necessary conformational changes required for function at the membrane. PMID:26133569

  17. Human immunodeficiency virus type 1 Nef-induced down-modulation of CD4 is due to rapid internalization and degradation of surface CD4.

    PubMed Central

    Rhee, S S; Marsh, J W

    1994-01-01

    Human immunodeficiency virus type 1 (HIV-1) Nef is a myristylated protein with a relative molecular mass of 27 kDa, is localized to the cytoplasmic surfaces of cellular membranes, and has been reported to down-modulate CD4 in human T cells. To understand the mechanism of HIV-1 Nef-mediated down-modulation of cell surface CD4, we expressed Nef protein in human T-cell line VB. Expression of HIV-1 Nef protein down-modulated surface CD4 molecules. In pulse-chase experiments, CD4 molecules in Nef-expressing cells were synthesized at normal levels. However, the bulk of newly synthesized CD4 protein was degraded with a half-life of approximately 6 h, compared with the 24-h half-life in control cells. This Nef-induced acceleration of CD4 turnover was inhibited by lysosomotropic agents NH4Cl and chloroquine as well as by the protease inhibitor leupeptin. Surface CD4 biotinylation experiments demonstrated that CD4 molecules in Nef-expressing T cells are transported to the plasma membrane with normal kinetics but are then rapidly internalized. Therefore, HIV-1 Nef-induced down-modulation of CD4 is due to rapid internalization of surface CD4 and subsequent degradation by an acid-dependent process, potentially lysosomal. Additionally, in a Nef-expressing cell, we find accelerated dissociation of the T-cell tyrosine kinase p56lck and CD4 but only after the complex reaches the plasma membrane. This implies that HIV-1 Nef protein might play a role in triggering a series of T-cell activation-like events, which contribute to p56lck dissociation and internalization of surface CD4 molecules. Images PMID:8035515

  18. Nef proteins from simian immunodeficiency viruses are tetherin antagonists

    PubMed Central

    Zhang, Fengwen; Wilson, Sam J.; Langford, Wilmina; Virgen, Beatriz; Gregory, Devon; Johnson, Marc; Munch, Jan; Kirchhoff, Frank; Bieniasz, Paul D.; Hatziioannou, Theodora

    2010-01-01

    The tetherin/BST2/CD317 protein blocks the release of HIV-1 and other enveloped viruses by inducing tethering of nascent particles to infected cell surfaces. The HIV-1 Vpu protein antagonizes the antiviral activity of human but not monkey tetherins and many simian immunodeficiency viruses (SIVs) do not encode Vpu. Here, we show that the apparently ‘missing’ anti-tetherin activity in SIVs has been acquired by several SIV Nef proteins. Specifically, SIVMAC/SIVSMM, SIVAGM and SIVBLU Nef proteins can suppress tetherin activity. Notably, tetherin antagonism by SIV Nef proteins is species-specific, is genetically separable from other Nef activities and is most evident with simian rather than human tetherin proteins. Accordingly, a critical determinant of sensitivity to SIVMAC Nef in the tetherin cytoplasmic tail is variable in nonhuman primate tetherins and deleted in human tetherin, likely due to selective pressures imposed by viral antagonists, perhaps including Nef proteins. PMID:19501037

  19. Optimized Replicating Renilla Luciferase Reporter HIV-1 Utilizing Novel Internal Ribosome Entry Site Elements for Native Nef Expression and Function.

    PubMed

    Alberti, Michael O; Jones, Jennifer J; Miglietta, Riccardo; Ding, Haitao; Bakshi, Rakesh K; Edmonds, Tara G; Kappes, John C; Ochsenbauer, Christina

    2015-12-01

    We previously developed replication-competent reporter HIV-1 (referred to herein as LucR.T2A reporter viruses), utilizing a "ribosome skipping" T2A peptide strategy to link Renilla luciferase (LucR) with Nef expression. The demonstrated utility for HIV-1 vaccine and transmission study applications included measurement of neutralizing antibody (NAb) activity in vaccine sera, improved cell-mediated virus inhibition assays, such as T cell-mediated virus inhibition and antibody-dependent cell-mediated cytotoxicity (ADCC) assays, and humanized mouse models. Herein, we extend our prior work and introduce reporter virus technology for applications that require fully functional Nef. We demonstrate that in CD4(+) T cells productively infected with LucR.T2A reporter viruses, T2A peptide-driven Nef expression and function, such as down-regulation of surface CD4 and MHC-I, were impaired. We overcame this limitation of LucR.T2A reporter viruses and achieved physiological Nef expression and function by engineering novel LucR reporter HIV-1 comprising 11 different internal ribosome entry site (IRES) elements chosen for size and relative activity. A range of Nef expression was observed in 293T cells transfected with the different LucR.IRES reporter virus constructs. Iteratively, we identified IRES reporter genomes that expressed Nef closest to physiological levels and produced virus with infectivity, titers, and replication kinetics similar to nonreporter viruses. Our results demonstrated that LucR reporter activity was stable over multiple replication cycles in peripheral blood mononuclear cells (PBMCs). Furthermore, we analyzed Nef functionality, i.e., down-modulation of MHC-I and CD4, following infection of T cell lines and PBMCs. Unlike LucR.T2A reporter virus, one of the redesigned LucR.IRES reporter viruses [containing the modified encephalomyocarditis virus (EMCV) 6ATR IRES element, "6ATRi"] demonstrated Nef expression and function similar to parental "nonreporter" virus

  20. Molecular characterization of HIV-1 Nef and ACOT8 interaction: insights from in silico structural predictions and in vitro functional assays

    NASA Astrophysics Data System (ADS)

    Serena, Michela; Giorgetti, Alejandro; Busato, Mirko; Gasparini, Francesca; Diani, Erica; Romanelli, Maria Grazia; Zipeto, Donato

    2016-03-01

    HIV-1 Nef interacts with several cellular proteins, among which the human peroxisomal thioesterase 8 (ACOT8). This interaction may be involved in the endocytosis regulation of membrane proteins and might modulate lipid composition in membrane rafts. Nef regions involved in the interaction have been experimentally characterized, whereas structural details of the ACOT8 protein are unknown. The lack of structural information hampers the comprehension of the functional consequences of the complex formation during HIV-1 infection. We modelled, through in silico predictions, the ACOT8 structure and we observed a high charge complementarity between Nef and ACOT8 surfaces, which allowed the identification of the ACOT8 putative contact points involved in the interaction. The predictions were validated by in vitro assays through the development of ACOT8 deletion mutants. Coimmunoprecipitation and immunofluorescence analyses showed that ACOT8 Arg45-Phe55 and Arg86-Pro93 regions are involved in Nef association. In addition, K91S mutation abrogated the interaction with Nef, indicating that Lys91 plays a key role in the interaction. Finally, when associated with ACOT8, Nef may be preserved from degradation. These findings improve the comprehension of the association between HIV-1 Nef and ACOT8, helping elucidating the biological effect of their interaction.

  1. Molecular characterization of HIV-1 Nef and ACOT8 interaction: insights from in silico structural predictions and in vitro functional assays

    PubMed Central

    Serena, Michela; Giorgetti, Alejandro; Busato, Mirko; Gasparini, Francesca; Diani, Erica; Romanelli, Maria Grazia; Zipeto, Donato

    2016-01-01

    HIV-1 Nef interacts with several cellular proteins, among which the human peroxisomal thioesterase 8 (ACOT8). This interaction may be involved in the endocytosis regulation of membrane proteins and might modulate lipid composition in membrane rafts. Nef regions involved in the interaction have been experimentally characterized, whereas structural details of the ACOT8 protein are unknown. The lack of structural information hampers the comprehension of the functional consequences of the complex formation during HIV-1 infection. We modelled, through in silico predictions, the ACOT8 structure and we observed a high charge complementarity between Nef and ACOT8 surfaces, which allowed the identification of the ACOT8 putative contact points involved in the interaction. The predictions were validated by in vitro assays through the development of ACOT8 deletion mutants. Coimmunoprecipitation and immunofluorescence analyses showed that ACOT8 Arg45-Phe55 and Arg86-Pro93 regions are involved in Nef association. In addition, K91S mutation abrogated the interaction with Nef, indicating that Lys91 plays a key role in the interaction. Finally, when associated with ACOT8, Nef may be preserved from degradation. These findings improve the comprehension of the association between HIV-1 Nef and ACOT8, helping elucidating the biological effect of their interaction. PMID:26927806

  2. Repair and evolution of nef in vivo modulates simian immunodeficiency virus virulence.

    PubMed Central

    Whatmore, A M; Cook, N; Hall, G A; Sharpe, S; Rud, E W; Cranage, M P

    1995-01-01

    Experimental evidence from the simian immunodeficiency virus (SIV) model of AIDS has shown that the nef gene is critical in the pathogenesis of AIDS. Consequently, nef is of considerable interest in both antiviral drug and vaccine development. Preliminary findings in two rhesus macaques indicated that a deletion of only 12 bp found in the overlapping nef/3' long terminal repeat (LTR) region (9501 to 9512) of the SIVmacC8 molecular clone was associated with reduced virus isolation frequency. We show that this deletion can be repaired in vivo by a sequence duplication event and that sequence evolution continues until the predicted amino acid sequence of the repair is virtually indistinguishable from that of the virulent wild type. These changes occurred concomitantly with reversion to virulence, evidenced by a high virus isolation frequency and load, decline in anti-p27 antibody, substantial reduction in the CD4/CD8 ratio, and development of opportunistic infections associated with AIDS. These findings clearly illustrate the capacity for repair of small attenuating deletions in primate lentiviruses and also strongly suggest that the region from 9501 to 9512 in the SIV nef/3' LTR region is of biological relevance. In addition, the ability of attenuated virus to revert to virulence raises fundamental questions regarding the nature of superinfection immunity. PMID:7609080

  3. Inhibition of human immunodeficiency virus type 1 by triciribine involves the accessory protein nef.

    PubMed

    Ptak, Roger G; Gentry, Brian G; Hartman, Tracy L; Watson, Karen M; Osterling, M Clayton; Buckheit, Robert W; Townsend, Leroy B; Drach, John C

    2010-04-01

    Triciribine (TCN) is a tricyclic nucleoside that inhibits human immunodeficiency virus type 1 (HIV-1) replication by a unique mechanism not involving the inhibition of enzymes directly involved in viral replication. This activity requires the phosphorylation of TCN to its 5' monophosphate by intracellular adenosine kinase. New testing with a panel of HIV and simian immunodeficiency virus isolates, including low-passage-number clinical isolates and selected subgroups of HIV-1, multidrug resistant HIV-1, and HIV-2, has demonstrated that TCN has broad antiretroviral activity. It was active in cell lines chronically infected with HIV-1 in which the provirus was integrated into chromosomal DNA, thereby indicating that TCN inhibits a late process in virus replication. The selection of TCN-resistant HIV-1 isolates resulted in up to a 750-fold increase in the level of resistance to the drug. DNA sequence analysis of highly resistant isolate HIV-1(H10) found five point mutations in the HIV-1 gene nef, resulting in five different amino acid changes. DNA sequencing of the other TCN-resistant isolates identified at least one and up to three of the same mutations observed in isolate HIV-1(H10). Transfer of the mutations from TCN-resistant isolate HIV-1(H10) to wild-type virus and subsequent viral growth experiments with increasing concentrations of TCN demonstrated resistance to the drug. We conclude that TCN is a late-phase inhibitor of HIV-1 replication and that mutations in nef are necessary and sufficient for TCN resistance.

  4. Rectal HSV-2 Infection May Increase Rectal SIV Acquisition Even in the Context of SIVΔnef Vaccination.

    PubMed

    Guerra-Pérez, Natalia; Aravantinou, Meropi; Veglia, Filippo; Goode, Diana; Truong, Rosaline; Derby, Nina; Blanchard, James; Grasperge, Brooke; Gettie, Agegnehu; Robbiani, Melissa; Martinelli, Elena

    2016-01-01

    Prevalent HSV-2 infection increases the risk of HIV acquisition both in men and women even in asymptomatic subjects. Understanding the impact of HSV-2 on the mucosal microenvironment may help to identify determinants of susceptibility to HIV. Vaginal HSV-2 infection increases the frequency of cells highly susceptible to HIV in the vaginal tissue of women and macaques and this correlates with increased susceptibility to vaginal SHIV infection in macaques. However, the effect of rectal HSV-2 infection on HIV acquisition remains understudied. We developed a model of rectal HSV-2 infection in macaques in combination with rectal SIVmac239Δnef (SIVΔnef) vaccination and our results suggest that rectal HSV-2 infection may increase the susceptibility of macaques to rectal SIVmac239 wild-type (wt) infection even in SIVΔnef-infected animals. Rectal SIVΔnef infection/vaccination protected 7 out of 7 SIVΔnef-infected macaques from SIVmac239wt rectal infection (vs 12 out of 16 SIVΔnef-negative macaques), while 1 out of 3 animals co-infected with SIVΔnef and HSV-2 acquired SIVmac239wt infection. HSV-2/SIVmac239wt co-infected animals had increased concentrations of inflammatory factors in their plasma and rectal fluids and a tendency toward higher acute SIVmac239wt plasma viral load. However, they had higher blood CD4 counts and reduced depletion of CCR5+ CD4+ T cells compared to SIVmac239wt-only infected animals. Thus, rectal HSV-2 infection generates a pro-inflammatory environment that may increase susceptibility to rectal SIV infection and may impact immunological and virological parameters during acute SIV infection. Studies with larger number of animals are needed to confirm these findings.

  5. Structural constraints on human immunodeficiency virus type 1 Nef function

    SciTech Connect

    Raney, Alexa; Shaw, Alice Y.; Foster, John L.; Garcia, J. Victor

    2007-11-10

    HIV-1 Nef is a multifunctional protein that exerts its activities through interactions with multiple cellular partners. Nef uses different domains and mechanisms to exert its functions including cell surface down-modulation of CD4 and MHC-I receptors and activation of the serine/threonine kinase PAK-2. We inserted tags at the C-terminus and proximal to the N-terminus of Nef and the effects on Nef's structure/function relationships were examined. We discovered significant defects in MHC-I down-modulation with the insertion of HA/FLAG tags at either region. We also found impaired PAK-2 activation with a C-terminal fusion with GFP. Interestingly, Nef-GFP and Nef-GH{sub 7} induced MHC-I down-modulation, suggesting that the negative charge of the HA/FLAG tag could contribute to the observed defect. Together, these observations highlight elements of Nef's functional complexity and demonstrate previously unsuspected structural requirements for PAK-2 activation and MHC-1 down-modulation in Nef's flexible N- and C-terminal regions.

  6. Human immunodeficiency virus type 1 nef quasispecies in pathological tissue.

    PubMed Central

    Blumberg, B M; Epstein, L G; Saito, Y; Chen, D; Sharer, L R; Anand, R

    1992-01-01

    The role of the nef gene in human immunodeficiency virus type 1 (HIV-1) infection is poorly understood. To provide a basis for studies on the role of nef in AIDS, we used targeted polymerase chain reaction amplification and DNA sequencing to determine the structure of nef genes in pathologic tissue from HIV-1-infected children and adults. We find that the nef reading frame is open in 92% of clones derived from both brain and lymphocytic tissue of children, suggesting that nef is expressed in these tissues. One HIV-1 clone, BRVA, obtained by coculture from the brain of an adult AIDS patient with progressive dementia, was previously shown to contain a duplicated region in nef. We show here that similar duplications are widespread in both adults and children with AIDS. However, coculture strongly selects against the broad spectrum of nef quasispecies found in tissue. These findings suggest functional selection for nef quasispecies in pathologic tissues during HIV-1 infection of the human host. Images PMID:1501274

  7. CD8 T Cell Response Maturation Defined by Anentropic Specificity and Repertoire Depth Correlates with SIVΔnef-induced Protection

    PubMed Central

    Adnan, Sama; Colantonio, Arnaud D.; Yu, Yi; Gillis, Jacqueline; Wong, Fay E.; Becker, Ericka A.; Reeves, R. Keith; Lifson, Jeffrey D.; O’Connor, Shelby L.; Johnson, R. Paul

    2015-01-01

    The live attenuated simian immunodeficiency virus (LASIV) vaccine SIVΔnef is one of the most effective vaccines in inducing protection against wild-type lentiviral challenge, yet little is known about the mechanisms underlying its remarkable protective efficacy. Here, we exploit deep sequencing technology and comprehensive CD8 T cell epitope mapping to deconstruct the CD8 T cell response, to identify the regions of immune pressure and viral escape, and to delineate the effect of epitope escape on the evolution of the CD8 T cell response in SIVΔnef-vaccinated animals. We demonstrate that the initial CD8 T cell response in the acute phase of SIVΔnef infection is mounted predominantly against more variable epitopes, followed by widespread sequence evolution and viral escape. Furthermore, we show that epitope escape expands the CD8 T cell repertoire that targets highly conserved epitopes, defined as anentropic specificity, and generates de novo responses to the escaped epitope variants during the vaccination period. These results correlate SIVΔnef-induced protection with expanded anentropic specificity and increased response depth. Importantly, these findings render SIVΔnef, long the gold standard in HIV/SIV vaccine research, as a proof-of-concept vaccine that highlights the significance of the twin principles of anentropic specificity and repertoire depth in successful vaccine design. PMID:25688559

  8. [Linking Nef with the immunological and virological determinants of AIDS].

    PubMed

    Gómez-Icazbalceta, Guillermo; Larralde, Carlos

    2009-01-01

    Nef is an accessory protein from HIV-1 involved in viral replication, depletion of CD4 + T cells and progression to AIDS. The participation of Nef in altering several of the cellular and subcellular components has been studied in detail. The finding that infected patients over 15 years with HIV-1 variants that expressed a defective nef gene had not progressed to AIDS, suggested that Nef played a decisive role in the outcome of AIDS, through the regulation of different virological and immunological components. In addition, the mere expression of Nef is able to induce AIDS in transgenic mice and alter the kinetics of replication of the virus in infected monkeys. This review examines the relationship between the expression of Nef and the ultimate consequences of HIV-1 infection, taking into account the most relevant clinical parameters: viral load, counting of CD4 + T cells and progression to AIDS. Under this approach, it is emphasized the role of Nef as immunomodulator and also as independent mediator of damage to the immune system.

  9. Mechanisms of HIV-1 Nef Function and Intracellular Signaling

    PubMed Central

    Foster, John L.; Denial, Sarah J.; Temple, Brenda R. S.; Garcia, J. Victor

    2013-01-01

    Advances in the last several years have enhanced mechanistic understanding of Nef induced CD4 and MHCI downregulation and have suggested a new paradigm for analyzing Nef function. In both of these cases, Nef acts by forming ternary complexes with significant contributions to stability imparted by non-canonical interactions. The mutational analyses and binding assays that have led to these conclusions are discussed. The recent progress has been dependent on conservative mutations and multi-protein binding assays. The poorly understood Nef functions of p21 activated protein kinase (PAK2) activation, enhancement of virion infectivity, and inhibition of immunoglobulin class switching are also likely to involve ternary complexes and non-canonical interactions. Hence, investigation of these latter Nef functions should benefit from a similar approach. Six historically used alanine substitutions for determining structure-function relationships of Nef are discussed. These are M20A, E62A/E63A/E64A/E65A (AAAA), P72A/P75A (AXXA), R106A, L164A/L165A, and D174A/D175A. Investigations of less disruptive mutations in place of AAAA and AXXA have led to different interpretations of mechanism. Two recent examples of this alternate approach applied to PAK2 activation F191 and critical residue D123 are presented. The implications of the new findings and the resulting new paradigm for Nef structure-function are discussed with respect to creating a map of Nef functions on the protein surface. We report the results of a PPI-Pred analysis for protein-protein interfaces. There are three predicted patches produced by the analysis which describe regions consistent with the currently known mutational analyses of Nef function. PMID:21336563

  10. The cytoplasmic domain of CD4 is sufficient for its down-regulation from the cell surface by human immunodeficiency virus type 1 Nef.

    PubMed Central

    Anderson, S J; Lenburg, M; Landau, N R; Garcia, J V

    1994-01-01

    Human immunodeficiency virus type 1 Nef down-regulates surface expression of murine and human CD4 but not human CD8. We recently reported that the cytoplasmic domain of CD4 is required for its down-regulation by Nef. Using a chimeric molecule composed of the extracellular and transmembrane domains of human CD8 fused to the cytoplasmic domain of human CD4, we show here that the cytoplasmic domain of CD4 is sufficient for down-regulation by Nef. Since the cytoplasmic domain of CD4 is also the site of its association with p56lck, we used a series of CD4 mutants to determine whether the regions of the cytoplasmic domain of CD4 required for down-regulation by Nef are the same as those required for p56lck binding. Our results indicate that the portion of the cytoplasmic domain required for the down-regulation of CD4 by Nef overlaps with the binding site of p56lck, but the cysteine residues which are essential for the association of CD4 with p56lck are not required. This observation raised the possibility that Nef competes with p56lck for binding to CD4. However, under conditions which are considerably milder than those permissive for coimmunoprecipitation of CD4 and p56lck, we found no evidence for an association between Nef and CD4. While a decrease in total CD4 was observed in lysates of cells expressing Nef, the levels of p56lck were not significantly affected. Pulse-chase experiments further revealed a decrease in the half-life of CD4 in Nef-expressing cells. These results show that the decrease in surface CD4 expression induced by Nef is mediated at least in part by a decrease in the half-life of CD4 protein. These results also indicate that a large portion of p56lck is free of CD4 in T cells expressing Nef, which could have a significant effect on T-cell function. Images PMID:8151774

  11. Immunogenic compositions comprising human immunodeficiency virus (HIV) mosaic Nef proteins

    DOEpatents

    Korber, Bette T [Los Alamos, NM; Perkins, Simon [Los Alamos, NM; Bhattacharya, Tanmoy [Los Alamos, NM; Fischer, William M [Los Alamos, NM; Theiler, James [Los Alamos, NM; Letvin, Norman [Boston, MA; Haynes, Barton F [Durham, NC; Hahn, Beatrice H [Birmingham, AL; Yusim, Karina [Los Alamos, NM; Kuiken, Carla [Los Alamos, NM

    2012-02-21

    The present invention relates to mosaic clade M HIV-1 Nef polypeptides and to compositions comprising same. The polypeptides of the invention are suitable for use in inducing an immune response to HIV-1 in a human.

  12. The human immunodeficiency virus antigen Nef forms protein bodies in leaves of transgenic tobacco when fused to zeolin.

    PubMed

    de Virgilio, Maddalena; De Marchis, Francesca; Bellucci, Michele; Mainieri, Davide; Rossi, Marika; Benvenuto, Eugenio; Arcioni, Sergio; Vitale, Alessandro

    2008-01-01

    Protein bodies (PB) are stable polymers naturally formed by certain seed storage proteins within the endoplasmic reticulum (ER). The human immunodeficiency virus negative factor (Nef) protein, a potential antigen for the development of an anti-viral vaccine, is highly unstable when introduced into the plant secretory pathway, probably because of folding defects in the ER environment. The aim of this study was to promote the formation of Nef-containing PB in tobacco (Nicotiana tabacum) leaves by fusing the Nef sequence to the N-terminal domains of the maize storage protein gamma-zein or to the chimeric protein zeolin (which efficiently forms PB and is composed of the vacuolar storage protein phaseolin fused to the N-terminal domains of gamma-zein). Protein blots and pulse-chase indicate that fusions between Nef and the same gamma-zein domains present in zeolin are degraded by ER quality control. Consistently, a mutated zeolin, in which wild-type phaseolin was substituted with a defective version known to be degraded by ER quality control, is unstable in plant cells. Fusion of Nef to the entire zeolin sequence instead allows the formation of PB detectable by electron microscopy and subcellular fractionation, leading to zeolin-Nef accumulation higher than 1% of total soluble protein, consistently reproduced in independent transgenic plants. It is concluded that zeolin, but not its gamma-zein portion, has a positive dominant effect over ER quality control degradation. These results provide insights into the requirements for PB formation and avoidance of quality-control degradation, and indicate a strategy for enhancing foreign protein accumulation in plants.

  13. The human immunodeficiency virus antigen Nef forms protein bodies in leaves of transgenic tobacco when fused to zeolin

    PubMed Central

    de Virgilio, Maddalena; Bellucci, Michele; Mainieri, Davide; Rossi, Marika; Benvenuto, Eugenio; Arcioni, Sergio; Vitale, Alessandro

    2008-01-01

    Protein bodies (PB) are stable polymers naturally formed by certain seed storage proteins within the endoplasmic reticulum (ER). The human immunodeficiency virus negative factor (Nef) protein, a potential antigen for the development of an anti-viral vaccine, is highly unstable when introduced into the plant secretory pathway, probably because of folding defects in the ER environment. The aim of this study was to promote the formation of Nef-containing PB in tobacco (Nicotiana tabacum) leaves by fusing the Nef sequence to the N-terminal domains of the maize storage protein γ-zein or to the chimeric protein zeolin (which efficiently forms PB and is composed of the vacuolar storage protein phaseolin fused to the N-terminal domains of γ-zein). Protein blots and pulse–chase indicate that fusions between Nef and the same γ-zein domains present in zeolin are degraded by ER quality control. Consistently, a mutated zeolin, in which wild-type phaseolin was substituted with a defective version known to be degraded by ER quality control, is unstable in plant cells. Fusion of Nef to the entire zeolin sequence instead allows the formation of PB detectable by electron microscopy and subcellular fractionation, leading to zeolin–Nef accumulation higher than 1% of total soluble protein, consistently reproduced in independent transgenic plants. It is concluded that zeolin, but not its γ-zein portion, has a positive dominant effect over ER quality control degradation. These results provide insights into the requirements for PB formation and avoidance of quality-control degradation, and indicate a strategy for enhancing foreign protein accumulation in plants. PMID:18540021

  14. Association between a naturally arising polymorphism within a functional region of HIV-1 Nef and disease progression in chronic HIV-1 infection.

    PubMed

    Meribe, Stanley C; Hasan, Zafrul; Mahiti, Macdonald; Mwimanzi, Francis; Toyoda, Mako; Mori, Masahiko; Gatanaga, Hiroyuki; Kikuchi, Tadashi; Miura, Toshiyuki; Kawana-Tachikawa, Ai; Iwamoto, Aikichi; Oka, Shinichi; Ueno, Takamasa

    2015-08-01

    HIV-1 Nef mediates downregulation of HLA class I (HLA-I) through a number of highly conserved sequence motifs. We investigated the in vivo implication(s) of naturally arising polymorphisms in functional motifs in HIV-1 Nef that are associated with HLA-I downregulation, including the acidic cluster, polyproline, di-arginine and Met-20 regions. Plasma samples from treatment-naive, chronically HIV-1 infected subjects were collected after obtaining informed consent, and viral RNA was extracted and amplified by nested RT-PCR. The resultant nef amplicons were sequenced directly, and subtype-B sequences with an intact open reading frame (n = 406) were included in our analyses. There was over-representation of isoleucine at position 20 (Ile-20) in our dataset when compared to sequences in the Los Alamos sequence database (17.7 vs. 6.9 %, p = 0.0309). The presence of having Ile-20 in Nef was found to be associated with higher median plasma viral load (p = 0.013), independent of associated codons or viral lineage effects, whereas no clinical association was found with polymorphisms in the other functional motifs. Moreover, introduction of a Met-20-to-Ile mutation in a laboratory strain SF2 Nef resulted in a modest, albeit not statistically significant, increase in HLA class I downregulation activity (p = 0.06). Taken together, we have identified a naturally arising polymorphism, Ile-20, within HIV-1 subtype B Nef that is associated with poorer disease outcome.

  15. Human Immunodeficiency Virus Type 1 Nef Inhibits Autophagy through Transcription Factor EB Sequestration

    PubMed Central

    Campbell, Grant R.; Rawat, Pratima; Bruckman, Rachel S.; Spector, Stephen A.

    2015-01-01

    HIV Nef acts as an anti-autophagic maturation factor through interaction with beclin-1 (BECN1). We report that exposure of macrophages to infectious or non-infectious purified HIV induces toll-like receptor 8 (TLR8) and BECN1 dependent dephosphorylation and nuclear translocation of TFEB and that this correlates with an increase in autophagy markers. RNA interference for ATG13, TFEB, TLR8, or BECN1 inhibits this HIV-induced autophagy. However, once HIV establishes a productive infection, TFEB phosphorylation and cytoplasmic sequestration are increased resulting in decreased autophagy markers. Moreover, by 7 d post-infection, autophagy levels are similar to mock infected controls. Conversely, although Nef deleted HIV similarly induces TFEB dephosphorylation and nuclear localization, and increases autophagy, these levels remain elevated during continued productive infection. Thus, the interaction between HIV and TLR8 serves as a signal for autophagy induction that is dependent upon the dephosphorylation and nuclear translocation of TFEB. During permissive infection, Nef binds BECN1 resulting in mammalian target of rapamycin (MTOR) activation, TFEB phosphorylation and cytosolic sequestration, and the inhibition of autophagy. To our knowledge, this is the first report of a virus modulating TFEB localization and helps to explain how HIV modulates autophagy to promote its own replication and cell survival. PMID:26115100

  16. Activation of the PAK-Related Kinase by Human Immunodeficiency Virus Type 1 Nef in Primary Human Peripheral Blood Lymphocytes and Macrophages Leads to Phosphorylation of a PIX-p95 Complex

    PubMed Central

    Brown, Amanda; Wang, Xia; Sawai, Earl; Cheng-Mayer, Cecilia

    1999-01-01

    Human immunodeficiency virus type 1 (HIV-1) Nef enhances virus replication in both primary T lymphocytes and monocyte-derived macrophages. This enhancement phenotype has been linked to the ability of Nef to modulate the activity of cellular kinases. We find that despite the reported high-affinity interaction between Nef and the Src kinase Hck in vitro, a Nef-Hck interaction in the context of HIV-1-infected primary macrophages is not detectable. However, Nef binding and activation of the PAK-related kinase and phosphorylation of its substrate could be readily detected in both infected primary T lymphocytes and macrophages. Furthermore, we show that this substrate is a complex composed of the recently characterized PAK interacting partner PIX (PAK-interacting guanine nucleotide exchange factor) and its tightly associated p95 protein. PAK and PIX-p95 appear to be differentially activated and phosphorylated depending on the intracellular environment in which nef is expressed. These results identify the PIX-p95 complex as a novel effector of Nef in primary cells and suggest that the regulation of the PAK signaling pathway may differ in T cells and macrophages. PMID:10559302

  17. Nef Binds p6* in GagPol during Replication of Human Immunodeficiency Virus Type 1

    PubMed Central

    Costa, Luciana J.; Zheng, Yong-Hui; Sabotic, Jerica; Mak, Johnson; Fackler, Oliver T.; Peterlin, B. Matija

    2004-01-01

    The atypical Nef protein (NefF12) from human immunodeficiency virus type 1 strain F12 (HIV-1F12) interferes with virion production and infectivity via a mysterious mechanism. The correlation of these effects with the unusual perinuclear subcellular localization of NefF12 suggested that the wild-type Nef protein could bind to assembly intermediates in late stages of viral replication. To test this hypothesis, Nef from HIV-1NL4-3 was fused to an endoplasmic reticulum (ER) retention signal (NefKKXX). This mutant NefKKXX protein recapitulated fully the effects of NefF12 on Gag processing and virion production, either alone or as a CD8 fusion protein. Importantly, the mutant NefKKXX protein also localized to the intermediate compartment, between the ER and the trans-Golgi network. Furthermore, Nef bound the GagPol polyprotein in vitro and in vivo. This binding mapped to the C-terminal flexible loop in Nef and the transframe p6* protein in GagPol. The significance of this interaction was demonstrated by a genetic assay in which the release of a mutant HIV-1 provirus lacking the PTAP motif in the late domain that no longer binds Tsg101 was rescued by a Nef.Tsg101 chimera. Importantly, this rescue as well as incorporation of Nef into HIV-1 virions correlated with the ability of Nef to interact with GagPol. Our data demonstrate that the retention of Nef in the intermediate compartment interferes with viral replication and suggest a new role for Nef in the production of HIV-1. PMID:15137387

  18. High level nuclear waste

    SciTech Connect

    Crandall, J L

    1980-01-01

    The DOE Division of Waste Products through a lead office at Savannah River is developing a program to immobilize all US high-level nuclear waste for terminal disposal. DOE high-level wastes include those at the Hanford Plant, the Idaho Chemical Processing Plant, and the Savannah River Plant. Commercial high-level wastes, for which DOE is also developing immobilization technology, include those at the Nuclear Fuel Services Plant and any future commercial fuels reprocessing plants. The first immobilization plant is to be the Defense Waste Processing Facility at Savannah River, scheduled for 1983 project submission to Congress and 1989 operation. Waste forms are still being selected for this plant. Borosilicate glass is currently the reference form, but alternate candidates include concretes, calcines, other glasses, ceramics, and matrix forms.

  19. Immune Selection In Vitro Reveals Human Immunodeficiency Virus Type 1 Nef Sequence Motifs Important for Its Immune Evasion Function In Vivo

    PubMed Central

    Lee, Patricia; Ng, Hwee L.; Yang, Otto O.

    2012-01-01

    Human immunodeficiency virus type 1 (HIV-1) Nef downregulates major histocompatibility complex class I (MHC-I), impairing the clearance of infected cells by CD8+ cytotoxic T lymphocytes (CTLs). While sequence motifs mediating this function have been determined by in vitro mutagenesis studies of laboratory-adapted HIV-1 molecular clones, it is unclear whether the highly variable Nef sequences of primary isolates in vivo rely on the same sequence motifs. To address this issue, nef quasispecies from nine chronically HIV-1-infected persons were examined for sequence evolution and altered MHC-I downregulatory function under Gag-specific CTL immune pressure in vitro. This selection resulted in decreased nef diversity and strong purifying selection. Site-by-site analysis identified 13 codons undergoing purifying selection and 1 undergoing positive selection. Of the former, only 6 have been reported to have roles in Nef function, including 4 associated with MHC-I downregulation. Functional testing of naturally occurring in vivo polymorphisms at the 7 sites with no previously known functional role revealed 3 mutations (A84D, Y135F, and G140R) that ablated MHC-I downregulation and 3 (N52A, S169I, and V180E) that partially impaired MHC-I downregulation. Globally, the CTL pressure in vitro selected functional Nef from the in vivo quasispecies mixtures that predominately lacked MHC-I downregulatory function at the baseline. Overall, these data demonstrate that CTL pressure exerts a strong purifying selective pressure for MHC-I downregulation and identifies novel functional motifs present in Nef sequences in vivo. PMID:22553319

  20. Chimeric human/murine monoclonal IgM antibodies to HIV-1 Nef antigen expressed on chronically infected cells.

    PubMed

    Kawai, Masahiro; He, Lianying; Kawamura, Takeshi; Omoto, Shinya; Fujii, Yoichi R; Okada, Noriko

    2003-01-01

    Human IgM antibody (Ab) to gangliosides induced cytolysis of HIV-1-infected cells by homologous human complement. We expected that any human IgM Ab reactive with HIV-1 infected cells could cause complement-mediated cytolysis. The trans-chromosome mouse (TC mouse) contains human chromosomes harboring genes responsible for immunoglobulin production. Spleen cells from TC mice immunized with recombinant Nef were fused with mouse myeloma cells to generate hybridomas, and we selected those that produced human mu-chain-positive Abs reactive with Nef fixed on an ELISA plate. However, the L-chain of the monoclonal Abs (mAbs) were murine lambda in type and were chimeric, and we could not succeed in obtaining mAb with human mu- and human kappa-chains. The chimeric mAbs reacted with the HIV-1 infected cells as seen with flow cytometric analysis, and the surface expression of Nef was also detectable on chronically infected OM10.1 cells which had no detectable gp120. However, although the reaction of the chimeric IgM mAb with HIV-1-infected MOLT4 cells induced C3 deposition on cell surfaces on incubation with fresh human serum, the cells remained unlysed, as determined by 51Cr release assay. The amount of Nef antigen on the cells might not have been high enough to overcome the function of HRF20 (CD59) that restricts formation of membrane attack complexes of homologous complement. However, combination of anti-Nef IgM mAb with other IgM mAbs reactive with the surface of HIV-1-infected cells may induce a synergistic effect in complement mediated cytolysis.

  1. Fusogenic activity of amino-terminal region of HIV type 1 Nef protein.

    PubMed

    Curtain, C C; Separovic, F; Rivett, D; Kirkpatrick, A; Waring, A J; Gordon, L M; Azad, A A

    1994-10-01

    We have studied two isoforms of Nef, Nef-27 and Nef-25, which were produced in E. coli. Nef-25 lacked the first 18 N-terminal residues of Nef-27 and both were nonmyristylated. Nef-27 fuses small unilamellar dipalmitoyl phosphatidylcholine vesicles (SUVs), as indicated by enhanced light scattering of SUVs and lipid mixing using concentration-dependent fluorescence dequenching. Nef-27 also causes the appearance of a shifted isotropic peak in the 31P NMR spectra of these vesicles, suggesting that protein interactions induce nonlamellar lipid structures. Recombinant Nef-25, which lacks only the 18 N-terminal residues of Nef-27, does not fuse vesicles and has little effect on the 31P NMR spectra. On the other hand, synthetic peptides consisting of 18 or 21 of the N-terminal residues of Nef-27 are strongly membrane perturbing, causing vesicle fusion and inducing isotropic peaks in the 31P NMR spectrum. Endogenous fluorescence spectra of the N-terminal peptide (21 residues) with SUVs show that the N-terminal sequence of Nef may achieve these perturbing effects by inserting its hydrophobic side into the lipid bilayer. Theoretical calculations using hydrophobic moment plot analysis indicate that short-length stretches (i.e., six amino acid residues) of the N-terminal sequence may insert into the lipid bilayer as multimeric alpha helices or beta sheets. The above-described membrane activities of Nef-27, which principally reside in its N-terminal domain, may play critical role(s) in certain functional properties of the full-length protein. For example, the fusogenic activity of the N-terminal sequence may be involved in the extracellular release of Nef-27, much of which appears to be associated with small membrane vesicles. The fusion activity may also be relevant to the ability of Nef-27 to downregulate CD4 and IL-2 receptors when this protein is electroporated into cultured lymphocytes, an activity not possessed by Nef-25.

  2. Effects of HIV-1 Nef on virus co-receptor expression and cytokine release in human bladder, laryngeal, and intestinal epithelial cell lines.

    PubMed

    Quaranta, Maria Giovanna; Falzano, Loredana; Vincentini, Olimpia; Fiorentini, Carla; Giordani, Luciana; Viora, Marina

    2011-06-01

    HIV infections are mainly acquired by mucosal transmission, through oral, rectal, or genital mucosa. Epithelial cells (EC) are the first cells encountered by HIV during infection through sexual transmission and breastfeeding. EC express several receptors critical for both primary HIV infection and secondary transmission. The regulation of co-receptor expression correlates with changes in susceptibility to infection by HIV-1 strains with different tropism. Moreover, inflammatory responses at mucosal surfaces after HIV-1 transmission may influence disease outcome. In the present study, we analyzed the effect of the accessory HIV-1 Nef protein on mucosal EC, using unstimulated or IFN-γ-stimulated HEp-2, T24, and Caco2 cell lines as models for homeostatic or inflamed mucosal tracts. We found that Nef significantly upregulated the expression of CXCR4 on the Caco-2 cell surface and the expression of galactosylceramide on the T24 cell surface. In addition, Nef significantly upregulated IL-6 production by T24 and Caco-2 cells, and TNF-α release by all three cell lines analyzed. Notably, Nef abrogated the IFN-γ-induced modulation of co-receptor expression and cytokine secretion. Our findings suggest that Nef differently regulates co-receptor expression and cytokine secretion at the epithelial level, depending on the anatomical derivation of the cells and the inflammatory status.

  3. Tracking the Emergence of Host-Specific Simian Immunodeficiency Virus env and nef Populations Reveals nef Early Adaptation and Convergent Evolution in Brain of Naturally Progressing Rhesus Macaques.

    PubMed

    Lamers, Susanna L; Nolan, David J; Rife, Brittany D; Fogel, Gary B; McGrath, Michael S; Burdo, Tricia H; Autissier, Patrick; Williams, Kenneth C; Goodenow, Maureen M; Salemi, Marco

    2015-08-01

    While a clear understanding of the events leading to successful establishment of host-specific viral populations and productive infection in the central nervous system (CNS) has not yet been reached, the simian immunodeficiency virus (SIV)-infected rhesus macaque provides a powerful model for the study of human immunodeficiency virus (HIV) intrahost evolution and neuropathogenesis. The evolution of the gp120 and nef genes, which encode two key proteins required for the establishment and maintenance of infection, was assessed in macaques that were intravenously inoculated with the same viral swarm and allowed to naturally progress to simian AIDS and potential SIV-associated encephalitis (SIVE). Longitudinal plasma samples and immune markers were monitored until terminal illness. Single-genome sequencing was employed to amplify full-length env through nef transcripts from plasma over time and from brain tissues at necropsy. nef sequences diverged from the founder virus faster than gp120 diverged. Host-specific sequence populations were detected in nef (~92 days) before they were detected in gp120 (~182 days). At necropsy, similar brain nef sequences were found in different macaques, indicating convergent evolution, while gp120 brain sequences remained largely host specific. Molecular clock and selection analyses showed weaker clock-like behavior and stronger selection pressure in nef than in gp120, with the strongest nef selection in the macaque with SIVE. Rapid nef diversification, occurring prior to gp120 diversification, indicates that early adaptation of nef in the new host is essential for successful infection. Moreover, the convergent evolution of nef sequences in the CNS suggests a significant role for nef in establishing neurotropic strains. The SIV-infected rhesus macaque model closely resembles HIV-1 immunopathogenesis, neuropathogenesis, and disease progression in humans. Macaques were intravenously infected with identical viral swarms to investigate

  4. A strategy of antigen incorporation into exosomes: comparing cross-presentation levels of antigens delivered by engineered exosomes and by lentiviral virus-like particles.

    PubMed

    Lattanzi, Laura; Federico, Maurizio

    2012-11-26

    Among strategies aimed at developing new nanoparticle-based vaccines, exosomes hold much promise. They are nanovesicles released by basically all eukaryotic cell types originating from intraluminal vesicles which accumulate in multivesicular bodies. Exosomes have immunogenic properties whose strength correlates with the amounts of associated antigens. Engineering antigens to target them in exosomes represents the last frontier in terms of nanoparticle-based vaccines. Here we report a new method to incorporate protein antigens in exosomes relying on the unique properties of a mutant of the HIV-1 Nef protein, Nef(mut). This is a biologically inactive mutant we found incorporating into exosomes at high levels also when fused at its C-terminus with foreign proteins. We compared both biochemical and antigenic properties of Nef(mut) exosomes with those of previously characterized Nef(mut) -based lentiviral virus-like particles (VLPs). We found that exosomes incorporate Nef(mut) and fusion protein derivatives with similar efficiency of VLPs. When an envelope fusion protein was associated with both exosomes and VLPs to favor cross-presentation of associated antigens, Nef(mut) and its derivatives incorporated in exosomes were cross-presented at levels at least similar to what observed when the antigens were delivered by engineered VLPs. This occurred despite exosomes entered target cells with an apparent lower efficiency than VLPs. The unique properties of HIV-1 Nef(mut) in terms of exosome incorporation efficiency, carrier of foreign antigens, and lack of anti-cellular effects open the way toward the development of a flexible, safe, cost-effective exosome-based CD8(+) T cell vaccine platform.

  5. Nef enhances HIV-1 infectivity via association with the virus assembly complex

    SciTech Connect

    Qi Mingli; Aiken, Christopher

    2008-04-10

    The HIV-1 accessory protein Nef enhances virus infectivity by facilitating an early post-entry step of infection. Nef acts in the virus producer cell, leading to a beneficial modification to HIV-1 particles. Nef itself is incorporated into HIV-1 particles, where it is cleaved by the viral protease during virion maturation. To probe the role of virion-associated Nef in HIV-1 infection, we generated a fusion protein consisting of the host protein cyclophilin A (CypA) linked to the amino terminus of Nef. The resulting CypA-Nef protein enhanced the infectivity of Nef-defective HIV-1 particles and was specifically incorporated into the virions via association with Gag during particle assembly. Pharmacologic or genetic inhibition of CypA-Nef binding to Gag prevented incorporation of CypA-Nef into virions and inhibited infectivity enhancement. Our results indicate that infectivity enhancement by Nef requires its association with a component of the assembling HIV-1 particle.

  6. Inhibition of Nef- and phorbol ester-induced CD4 degradation by macrolide antibiotics.

    PubMed Central

    Luo, T; Anderson, S J; Garcia, J V

    1996-01-01

    Human immunodeficiency virus type 1 (HIV-1) is the causative agent of AIDS. The simian immunodeficiency virus (SIV) causes a similar syndrome in macaques. The product of the nef gene of SIV has been shown to be important for virus replication and disease progression in vivo. In vitro, both SIV and HIV Nef downregulate surface expression of CD4 and accelerate total CD4 turnover. The mechanism by which Nef downregulates CD4 has not been established. A current model suggests that Nef enhances cell surface CD4 endocytosis and degradation in lysosomes. However, this was recently challenged when CD4 was found to accumulate in early endosomes of cells expressing Nef. Because inhibition of Nef function might halt virus replication and disease progression, we tested two macrolide antibiotics for their ability to inhibit Nef function. Concanamycin B (ConB) and bafilomycin A1 (BFLA1) are specific inhibitors of acidification of cell endosomes and lysosomes and, unlike other inhibitors, do not affect transport. Although ConB (25 nM) and BFLA1 (100 nM) blocked phorbol myristate acetate- and Nef-induced CD4 degradation in human monocyte U937 cells, CD4 surface expression was not recovered. Instead, CD4 accumulated in lysosomes. To determine if Nef is directly responsible for CD4 degradation or if they bind to each other in a manner similar to Vpu, transcripts of human CD4 and HIV-1 nef were cotranslated in vitro. Our results indicate that under our experimental conditions, Nef does not affect CD4 stability and does not associate with CD4 in this in vitro system. Our data suggest that (i) CD4 downregulation by Nef results in degradation of CD4 in lysosomes, (ii) inhibition of CD4 degradation by macrolide antibiotics does not restore surface expression, and (iii) the inhibition of CD4 expression by Nef appears to be indirect and is likely to involve cellular factors. PMID:8627671

  7. Cytolytic T lymphocytes (CTLs) from HIV-1 subtype C-infected Indian patients recognize CTL epitopes from a conserved immunodominant region of HIV-1 Gag and Nef.

    PubMed

    Thakar, Madhuri R; Bhonge, Leena S; Lakhashe, Samir K; Shankarkumar, U; Sane, Suvarna S; Kulkarni, Smita S; Mahajan, Bharati A; Paranjape, Ramesh S

    2005-09-01

    Analysis of the human immunodeficiency virus type 1 (HIV-1) cytolytic T lymphocyte (CTL) epitopes recognized by the targeted population is critical for HIV-1 vaccine design. Peripheral blood mononuclear cells from 47 Indian subjects at different stages of HIV-1 infection were tested for HIV-1 Gag-, Nef-, and Env-specific T cell responses by interferon (IFN)- gamma enzyme-linked immunospot (ELISPOT) assay, using pools of overlapping peptides. The Gag and Nef antigens were targeted by 83% and 36% of responders. Five immunodominant regions, 4 in Gag and 1 in Nef, were identified in the study; these regions are conserved across clades, including the African subtype C clade. Three antigenic regions were also found to be recognized by CTLs of the study participants. These regions were not identified as immunodominant regions in studies performed in Africa, which highlights the importance of differential clustering of responses within HIV-1 subtype C. Twenty-six putative epitopes--15 Gag (10 in p24 and 5 in p17), 10 Nef, and 1 Env (gp 41)--were predicted using a combination of peptide matrix ELISPOT assay and CTL epitope-prediction software. Ninety percent of the predicted epitopes were clustered in the conserved immunodominant regions of the Gag and Nef antigens. Of 26 predicted epitopes, 8 were promiscuous, 3 of which were highly conserved across clades. Three Gag and 4 Nef epitopes were novel. The identification of conserved epitopes will be important in the planning of an HIV-1 vaccine strategy for subtype C-affected regions.

  8. Virological and Immunological Characterization of Novel NYVAC-Based HIV/AIDS Vaccine Candidates Expressing Clade C Trimeric Soluble gp140(ZM96) and Gag(ZM96)-Pol-Nef(CN54) as Virus-Like Particles

    PubMed Central

    Perdiguero, Beatriz; Gómez, Carmen Elena; Cepeda, Victoria; Sánchez-Sampedro, Lucas; García-Arriaza, Juan; Mejías-Pérez, Ernesto; Jiménez, Victoria; Sánchez, Cristina; Sorzano, Carlos Óscar S.; Oliveros, Juan Carlos; Delaloye, Julie; Roger, Thierry; Calandra, Thierry; Asbach, Benedikt; Wagner, Ralf; Kibler, Karen V.; Jacobs, Bertram L.; Pantaleo, Giuseppe

    2014-01-01

    (CN54) as VLPs. These vectors are stable and express high levels of both HIV-1 antigens. Gag-induced VLPs do not interfere with NYVAC morphogenesis, are highly attenuated in immunocompromised and newborn mice after intracranial inoculation, trigger specific innate immune responses in human cells, and activate T (Env-specific CD4 and Gag-specific CD8) and B cell immune responses to the HIV antigens, leading to high antibody titers against gp140. For these reasons, these vectors can be considered vaccine candidates against HIV/AIDS and currently are being tested in macaques and humans. PMID:25355891

  9. Hydrogen Exchange Mass Spectrometry of Related Proteins with Divergent Sequences: A Comparative Study of HIV-1 Nef Allelic Variants

    NASA Astrophysics Data System (ADS)

    Wales, Thomas E.; Poe, Jerrod A.; Emert-Sedlak, Lori; Morgan, Christopher R.; Smithgall, Thomas E.; Engen, John R.

    2016-06-01

    Hydrogen exchange mass spectrometry can be used to compare the conformation and dynamics of proteins that are similar in tertiary structure. If relative deuterium levels are measured, differences in sequence, deuterium forward- and back-exchange, peptide retention time, and protease digestion patterns all complicate the data analysis. We illustrate what can be learned from such data sets by analyzing five variants (Consensus G2E, SF2, NL4-3, ELI, and LTNP4) of the HIV-1 Nef protein, both alone and when bound to the human Hck SH3 domain. Regions with similar sequence could be compared between variants. Although much of the hydrogen exchange features were preserved across the five proteins, the kinetics of Nef binding to Hck SH3 were not the same. These observations may be related to biological function, particularly for ELI Nef where we also observed an impaired ability to downregulate CD4 surface presentation. The data illustrate some of the caveats that must be considered for comparison experiments and provide a framework for investigations of other protein relatives, families, and superfamilies with HX MS.

  10. Expression and biochemical characterization of human immunodeficiency virus type 1 nef gene product.

    PubMed Central

    Kaminchik, J; Bashan, N; Pinchasi, D; Amit, B; Sarver, N; Johnston, M I; Fischer, M; Yavin, Z; Gorecki, M; Panet, A

    1990-01-01

    nef genes from human immunodeficiency virus type 1 isolates BH10 and LAV1 (lymphadenopathy-associated virus type 1) were expressed in Escherichia coli under the deo operon promoter. The two proteins found in the soluble compartment of the bacterial lysate were purified by ion-exchange column chromatography to apparent homogeneity. Determination of the amino-terminal sequence revealed glycine as the first amino acid in the Nef protein, indicating removal of the initiator methionine during expression in E. coli. Under native conditions, the recombinant Nef protein is a monomer of 23 kilodaltons. In denaturing polyacrylamide gels, however, BH10 and LAV1 Nef proteins migrate as 28 and 26 kilodaltons, respectively. GTP binding and GTPase activity were monitored during Nef protein purification. These activities did not copurify with the recombinant Nef protein from either the BH10 or the LAV1 isolate. Purified recombinant BH10 Nef protein was used as an immunogen to elicit mouse monoclonal antibodies. A series of monoclonal antibodies were obtained which reacted with sequences at either the amino or carboxy terminus of Nef. In addition, a conformational epitope reacting with native BH10, but not LAV1, Nef was isolated. Images PMID:2191151

  11. Interactions between Nef and AIP1 proliferate multivesicular bodies and facilitate egress of HIV-1

    PubMed Central

    Costa, Luciana J; Chen, Nan; Lopes, Adriana; Aguiar, Renato S; Tanuri, Amilcar; Plemenitas, Ana; Peterlin, B Matija

    2006-01-01

    Background Nef is an accessory protein of primate lentiviruses, HIV-1, HIV-2 and SIV. Besides removing CD4 and MHC class I from the surface and activating cellular signaling cascades, Nef also binds GagPol during late stages of the viral replicative cycle. In this report, we investigated further the ability of Nef to facilitate the replication of HIV-1. Results To this end, first the release of new viral particles was much lower in the absence of Nef in a T cell line. Since the same results were obtained in the absence of the viral envelope using pseudo-typed viruses, this phenomenon was independent of CD4 and enhanced infectivity. Next, we found that Nef not only possesses a consensus motif for but also binds AIP1 in vitro and in vivo. AIP1 is the critical intermediate in the formation of multivesicular bodies (MVBs), which play an important role in the budding and release of viruses from infected cells. Indeed, Nef proliferated MVBs in cells, but only when its AIP1-binding site was intact. Finally, these functions of Nef were reproduced in primary macrophages, where the wild type but not mutant Nef proteins led to increased release of new viral particles from infected cells. Conclusion We conclude that by binding GagPol and AIP1, Nef not only proliferates MVBs but also contributes to the egress of viral particles from infected cells. PMID:16764724

  12. Astrocytic expression of HIV-1 Nef impairs spatial and recognition memory

    PubMed Central

    Chompre, Gladys; Cruz, Emmanuel; Maldonado, Lucianette; Rivera-Amill, Vanessa; Porter, James T.; Noel, Richard J.

    2012-01-01

    Despite the widespread use of antiretroviral therapy that effectively limits viral replication, memory impairment remains a dilemma for HIV infected people. In the CNS, HIV infection of astrocytes leads to the production of the HIV-1 Nef protein without viral replication. Post mortem studies have found Nef expression in hippocampal astrocytes of people with HIV associated dementia suggesting that astrocytic Nef may contribute to HIV associated cognitive impairment even when viral replication is suppressed. To test whether astrocytic expression of Nef is sufficient to induce cognitive deficits, we examined the effect of implanting primary rat astrocytes expressing Nef into the hippocampus on spatial and recognition memory. Rats implanted unilaterally with astrocytes expressing Nef showed impaired novel location and novel object recognition in comparison with controls implanted with astrocytes expressing green fluorescent protein (GFP). This impairment was correlated with an increase in chemokine ligand 2 (CCL2) expression and the infiltration of peripheral macrophages into the hippocampus at the site of injection. Furthermore, the Nef exposed rats exhibited a bilateral loss of CA3 neurons. These results suggest that Nef protein expressed by the implanted astrocytes activates the immune system leading to neuronal damage and spatial and recognition memory deficits. Therefore, the continued expression of Nef by astrocytes in the absence of viral replication has the potential to contribute to HIV associated cognitive impairment. PMID:22926191

  13. D186/D190 is an allele-dependent determinant of HIV-1 Nef function.

    PubMed

    Imle, Andrea; Stolp, Bettina; Böhmer, Verena; Geyer, Matthias; Raz, Erez; Fackler, Oliver T

    2016-11-01

    The HIV-1 pathogenesis factor Nef interacts with numerous ligands to affect cellular vesicular transport, signal transduction and cytoskeletal dynamics. While most Nef functions depend on multivalent protein interaction motifs, disrupting actin dynamics requires a motif that specifically recruits the host kinase PAK2. An adjacent aspartate was recently predicted to mediate Nef-β-catenin interactions. We report here that β-catenin can be co-immunoprecipitated with Nef.GFP from Jurkat T cell lysates. This association is conserved among lentiviral Nef proteins but does not involve classical Nef protein interaction motifs, including the critical aspartate. While aspartate-to-alanine mutations impaired cell surface receptor downregulation and interference with actin dynamics and cell motility by HIV-1 NA7 Nef, analogous mutations did not affect HIV-1 SF2 Nef function. These allelic differences were determined by a proximal lysine/arginine polymorphism. These results emphasize differences between Nef alleles regarding the functional role of individual residues and underscore the need for allele-specific structure-function analyses. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. The human immunodeficiency virus (HIV) Rev-binding protein (HRB) is a co-factor for HIV-1 Nef-mediated CD4 downregulation.

    PubMed

    Landi, Alessia; Timermans, Cristina Garcia; Naessens, Evelien; Vanderstraeten, Hanne; Stove, Veronique; Verhasselt, Bruno

    2016-03-01

    Human immunodeficiency virus type 1 (HIV-1)-mediated CD4 downregulation is an important determinant of viral replication in vivo. Research on cellular co-factors involved in this process could lead to the identification of potential therapeutic targets. We found that CD4 surface levels were significantly higher in HIV-1-infected cells knocked-down for the HIV Rev-binding protein (HRB) compared with control cells. HRB knock-down affected CD4 downregulation induced by Nef but not by HIV-1 Vpu. Interestingly, the knock-down of the related protein HRBL (HRB-like), but not of the HRB interaction partner EPS15 (epidermal growth factor receptor pathway substrate 15), increased CD4 levels in Vpu-expressing cells significantly. Both of these proteins are known to be involved in HIV-1-mediated CD4 downregulation as co-factors of HIV-1 Nef. These results identify HRB as a previously unknown co-factor for HIV-1 Nef-mediated CD4 downregulation and highlight differences with the related protein HRBL, which affects the CD4 downregulation in a dual role as co-factor of both HIV-1 Nef and Vpu.

  15. Activation of NFAT-Dependent Gene Expression by Nef: Conservation among Divergent Nef Alleles, Dependence on SH3 Binding and Membrane Association, and Cooperation with Protein Kinase C-θ

    PubMed Central

    Manninen, Aki; Huotari, Päivi; Hiipakka, Marita; Renkema, G. Herma; Saksela, Kalle

    2001-01-01

    Here we show that the potential to regulate NFAT is a conserved property of different Nef alleles and that Nef residues involved in membrane targeting and SH3 binding are critical for this function. Cotransfection of an activated protein kinase C-θ (PKC-θ) with Nef implicated PKC-θ as a possible physiological cofactor of Nef in promoting NFAT-dependent gene expression and T-cell activation. PMID:11222731

  16. HIV-1 Nef binds with human GCC185 protein and regulates mannose 6 phosphate receptor recycling

    SciTech Connect

    Kumar, Manjeet; Kaur, Supinder; Nazir, Aamir; Tripathi, Raj Kamal

    2016-05-20

    HIV-1 Nef modulates cellular function that enhances viral replication in vivo which culminate into AIDS pathogenesis. With no enzymatic activity, Nef regulates cellular function through host protein interaction. Interestingly, trans-cellular introduction of recombinant Nef protein in Caenorhabditis elegans results in AIDS like pathogenesis which might share common pathophysiology because the gene sequence of C. elegans and humans share considerable homology. Therefore employing C. elegans based initial screen complemented with sequence based homology search we identified GCC185 as novel host protein interacting with HIV-1 Nef. The detailed molecular characterization revealed N-terminal EEEE{sub 65} acidic domain of Nef as key region for interaction. GCC185 is a tethering protein that binds with Rab9 transport vesicles. Our results show that Nef-GCC185 interaction disrupts Rab9 interaction resulting in delocalization of CI-MPR (cation independent Mannose 6 phosphate receptor) resulting in elevated secretion of hexosaminidase. In agreement with this, our studies identified novel host GCC185 protein that interacts with Nef EEEE65 acidic domain interfering GCC185-Rab9 vesicle membrane fusion responsible for retrograde vesicular transport of CI-MPR from late endosomes to TGN. In light of existing report suggesting critical role of Nef-GCC185 interaction reveals valuable mechanistic insights affecting specific protein transport pathway in docking of late endosome derived Rab9 bearing transport vesicle at TGN elucidating role of Nef during viral pathogenesis. -- Highlights: •Nef, an accessory protein of HIV-1 interacts with host factor and culminates into AIDS pathogenesis. •Using Caenorhabditis elegans based screen system, novel Nef interacting cellular protein GCC185 was identified. •Molecular characterization of Nef and human protein GCC185 revealed Nef EEEE{sub 65} key region interacted with full length GCC185. •Nef impeded the GCC185-Rab 9 interaction and

  17. Construction of Nef-positive doxycycline-dependent HIV-1 variants using bicistronic expression elements

    SciTech Connect

    Velden, Yme U. van der; Kleibeuker, Wendy; Harwig, Alex; Klaver, Bep; Siteur-van Rijnstra, Esther; Frankin, Esmay; Berkhout, Ben; Das, Atze T.

    2016-01-15

    Conditionally replicating HIV-1 variants that can be switched on and off at will are attractive tools for HIV research. We previously developed a genetically modified HIV-1 variant that replicates exclusively when doxycycline (dox) is administered. The nef gene in this HIV-rtTA variant was replaced with the gene encoding the dox-dependent rtTA transcriptional activator. Because loss of Nef expression compromises virus replication in primary cells and precludes studies on Nef function, we tested different approaches to restore Nef production in HIV-rtTA. Strategies that involved translation via an EMCV or synthetic internal ribosome entry site (IRES) failed because these elements were incompatible with efficient virus replication. Fusion protein approaches with the FMDV 2A peptide and human ubiquitin were successful and resulted in genetically-stable Nef-expressing HIV-rtTA strains that replicate more efficiently in primary T-cells and human immune system (HIS) mice than Nef-deficient variants, thus confirming the positive effect of Nef on in vivo virus replication. - Highlights: • Different approaches to encode additional proteins in the HIV-1 genome were tested. • IRES translation elements are incompatible with efficient HIV-1 replication. • Ubiquitin and 2A fusion protein approaches allow efficient HIV-1 replication. • Doxycycline-controlled HIV-1 variants that encode all viral proteins were developed. • Nef stimulates HIV-rtTA replication in primary cells and human immune system mice.

  18. Structural basis of evasion of cellular adaptive immunity by HIV-1 Nef

    SciTech Connect

    Jia, Xiaofei; Singh, Rajendra; Homann, Stefanie; Yang, Haitao; Guatelli, John; Xiong, Yong

    2012-10-24

    The HIV-1 protein Nef inhibits antigen presentation by class I major histocompatibility complex (MHC-I). We determined the mechanism of this activity by solving the crystal structure of a protein complex comprising Nef, the MHC-I cytoplasmic domain (MHC-I CD) and the {mu}1 subunit of the clathrin adaptor protein complex 1. A ternary, cooperative interaction clamps the MHC-I CD into a narrow binding groove at the Nef-{mu}1 interface, which encompasses the cargo-recognition site of {mu}1 and the proline-rich strand of Nef. The Nef C terminus induces a previously unobserved conformational change in {mu}1, whereas the N terminus binds the Nef core to position it optimally for complex formation. Positively charged patches on {mu}1 recognize acidic clusters in Nef and MHC-I. The structure shows how Nef functions as a clathrin-associated sorting protein to alter the specificity of host membrane trafficking and enable viral evasion of adaptive immunity.

  19. Synthesis and evaluation of orally active small molecule HIV-1 Nef antagonists.

    PubMed

    Emert-Sedlak, Lori A; Loughran, H Marie; Shi, Haibin; Kulp, John L; Shu, Sherry T; Zhao, Jielu; Day, Billy W; Wrobel, Jay E; Reitz, Allen B; Smithgall, Thomas E

    2016-03-01

    The HIV-1 Nef accessory factor enhances viral replication and promotes immune system evasion of HIV-infected cells, making it an attractive target for drug discovery. Recently we described a novel class of diphenylpyrazolodiazene compounds that bind directly to Nef in vitro and inhibit Nef-dependent HIV-1 infectivity and replication in cell culture. However, these first-generation Nef antagonists have several structural liabilities, including an azo linkage that led to poor oral bioavailability. The azo group was therefore replaced with either a one- or two-carbon linker. The resulting set of non-azo analogs retained nanomolar binding affinity for Nef by surface plasmon resonance, while inhibiting HIV-1 replication with micromolar potency in cell-based assays without cytotoxicity. Computational docking studies show that these non-azo analogs occupy the same predicted binding site within the HIV-1 Nef dimer interface as the original azo compound. Computational methods also identified a hot spot for inhibitor binding within this site that is defined by conserved HIV-1 Nef residues Asp108, Leu112, and Pro122. Pharmacokinetic evaluation of the non-azo B9 analogs in mice showed that replacement of the azo linkage dramatically enhanced oral bioavailability without substantially affecting plasma half-life or clearance. The improved oral bioavailability of non-azo diphenylpyrazolo Nef antagonists provides a starting point for further drug lead optimization in support of future efficacy testing in animal models of HIV/AIDS.

  20. Biochemical and biological comparison of HIV-1 NEF and ras gene products.

    PubMed

    Nebreda, A R; Bryan, T; Segade, F; Wingfield, P; Venkatesan, S; Santos, E

    1991-07-01

    Human immunodeficiency virus type 1 (HIV-1) NEF protein has been reported to share certain biochemical and structural properties with known oncoproteins like src or rats. To determine whether this is a general property of NEF from various HIV isolates, three different NEF proteins were expressed in Escherichia coli using a thermoinducible expression system previously exploited to overproduce functionally active p21 ras proteins. ras and NEF proteins expressed in this manner were evaluated in parallel to compare their biochemical and biological properties. In contrast to ras, our NEF protein preparations had no detectable GTP binding but showed autophosphorylation activity when incubated in the presence of either GTP or ATP. This putative autokinase activity was higher in NEF proteins containing threonine at position 15 than in those carrying alanine at that position. Two different NEF genes also failed to induce oncogenic transformation of permanently transfected NIH 3T3 cells under conditions that led to oncogenic transformation using activated ras genes. Also, unlike ras, the NEF gene products failed to induce meiotic maturation when injected into fully grown Xenopus oocytes.

  1. How HIV-1 Nef hijacks the AP-2 clathrin adaptor to downregulate CD4.

    PubMed

    Ren, Xuefeng; Park, Sang Yoon; Bonifacino, Juan S; Hurley, James H

    2014-01-01

    The Nef protein of HIV-1 downregulates the cell surface co-receptor CD4 by hijacking the clathrin adaptor complex AP-2. The structural basis for the hijacking of AP-2 by Nef is revealed by a 2.9 Å crystal structure of Nef bound to the α and σ2 subunits of AP-2. Nef binds to AP-2 via its central loop (residues 149-179) and its core. The determinants for Nef binding include residues that directly contact AP-2 and others that stabilize the binding-competent conformation of the central loop. Residues involved in both direct and indirect interactions are required for the binding of Nef to AP-2 and for downregulation of CD4. These results lead to a model for the docking of the full AP-2 tetramer to membranes as bound to Nef, such that the cytosolic tail of CD4 is situated to interact with its binding site on Nef. DOI: http://dx.doi.org/10.7554/eLife.01754.001.

  2. HIV-1 Nef associates with p22-phox, a component of the NADPH oxidase protein complex.

    PubMed

    Salmen, Siham; Colmenares, Melisa; Peterson, Darrel L; Reyes, Elbert; Rosales, Jose D; Berrueta, Lisbeth

    2010-01-01

    Altered neutrophil function may contribute to the development of AIDS during the course of HIV infection. It has been described that Nef, a regulatory protein from HIV, can modulate superoxide production in other cells, therefore altered superoxide production in neutrophils from HIV infected patients, could be secondary to a direct effect of Nef on components of the NADPH oxidase complex. In this work, we describe that Nef, was capable of increasing superoxide production in human neutrophils. Furthermore, a specific association between Nef and p22-phox, a membrane component of the NADPH oxidase complex, was found. We propose that this association may reflect a capability of Nef to modulate by direct association, the enzymatic complex responsible for one of the most efficient innate defense mechanisms in phagocytes, contributing to the pathogenesis of the disease. Copyright (c) 2010 Elsevier Inc. All rights reserved.

  3. Nef Proteins of Epidemic HIV-1 Group O Strains Antagonize Human Tetherin

    PubMed Central

    Kluge, Silvia F.; Mack, Katharina; Iyer, Shilpa S.; Pujol, François M.; Heigele, Anke; Learn, Gerald H.; Usmani, Shariq M.; Sauter, Daniel; Joas, Simone; Hotter, Dominik; Bibollet-Ruche, Frederic; Plenderleith, Lindsey J.; Peeters, Martine; Geyer, Matthias; Sharp, Paul M.; Fackler, Oliver T.; Hahn, Beatrice H.; Kirchhoff, Frank

    2014-01-01

    SUMMARY Most simian immunodeficiency viruses use their Nef protein to antagonize the host restriction factor tetherin. A deletion in human tetherin confers Nef resistance, representing a hurdle to successful zoonotic transmission. HIV-1 group M evolved to utilize the viral protein U (Vpu) to counteract tetherin. Although HIV-1 group O has spread epidemically in humans, it has not evolved a Vpu-based tetherin antagonism. Here we show that HIV-1 group O Nef targets a region adjacent to this deletion to inhibit transport of human tetherin to the cell surface, enhances virion release, and increases viral resistance to inhibition by interferon-α. The Nef protein of the inferred common ancestor of group O viruses is also active against human tetherin. Thus, Nef-mediated antagonism of human tetherin evolved prior to the spread of HIV-1 group O and likely facilitated secondary virus transmission. Our results may explain the epidemic spread of HIV-1 group O. PMID:25525794

  4. Is the high virulence of HIV-1 an unfortunate coincidence of primate lentiviral evolution?

    PubMed

    Kirchhoff, Frank

    2009-06-01

    In the subset of primate lentiviruses that contain a vpu gene - HIV-1 and its simian precursors - the Nef protein has lost the ability to down-modulate CD3, block T cell activation and suppress programmed death. Vpu counteracts a host restriction factor induced by the inflammatory cytokine interferon-alpha. I propose that the acquisition of vpu may have allowed the viral lineage that gave rise to HIV-1 to evolve towards greater pathogenicity by removing the selective pressure for a protective Nef function that prevents damagingly high levels of immune activation.

  5. Characterization of Three nef-Defective Human Immunodeficiency Virus Type 1 Strains Associated with Long-Term Nonprogression

    PubMed Central

    Rhodes, David I.; Ashton, Lesley; Solomon, Ajantha; Carr, Andrew; Cooper, David; Kaldor, John; Deacon, Nicholas

    2000-01-01

    Long-term survivors (LTS) of human immunodeficiency virus type 1 (HIV-1) infection provide an opportunity to investigate both viral and host factors that influence the rate of disease progression. We have identified three HIV-1-infected individuals in Australia who have been infected for over 11 years with viruses that contain deletions in the nef and nef-long terminal repeat (nef/LTR) overlap regions. These viruses differ from each other and from other nef-defective strains of HIV-1 previously identified in Australia. One individual, LTS 3, is infected with a virus containing a nef gene with a deletion of 29 bp from the nef/LTR overlap region, resulting in a truncated Nef open reading frame. In addition to the Nef defect, only viruses containing truncated Vif open reading frames of 37 or 69 amino acids could be detected in peripheral blood mononuclear cells isolated from this patient. LTS 3 had a viral load of less than 20 copies of RNA/ml of plasma. The other two long-term survivors, LTS 9 and LTS 11, had loads of less than 200 copies of RNA/ml of plasma and are infected with viruses with larger deletions in both the nef alone and nef/LTR overlap regions. These viruses contain wild-type vif, vpu, and vpr accessory genes. All three strains of virus had envelope sequences characteristic of macrophagetropic viruses. These findings further indicate the reduced pathogenic potential of nef-defective viruses. PMID:11044102

  6. Extracellular Nef Protein Targets CD4+ T Cells for Apoptosis by Interacting with CXCR4 Surface Receptors

    PubMed Central

    James, Cleve O.; Huang, Ming-Bo; Khan, Mafuz; Garcia-Barrio, Minerva; Powell, Michael D.; Bond, Vincent C.

    2004-01-01

    The effects of soluble Nef protein on CD4+ T cells were examined. CD4+-T-cell cultures exposed to soluble Nef were analyzed for apoptosis by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling and hallmarks of apoptosis including cytoplasmic shrinkage, nuclear fragmentation, DNA laddering, and caspase activation. We observed dose- and time-dependent inductions of apoptosis. DNA laddering and activated caspase 3 were also evident. Cells treated with Nef/protein kinase inhibitor complexes were protected from Nef-induced apoptosis, suggesting possible roles for protein kinases in the apoptosis pathway. Similarly, cells treated with Nef/anti-Nef antibody complexes were protected from Nef-induced apoptosis. The cellular receptor responsible for Nef-induced apoptosis was identified through antibody- and ligand-blocking experiments as a receptor commonly involved in viral entry. CXCR4 antibodies, as well as the endogenous ligand SDF-1α, were effective in blocking Nef-induced apoptosis, while CCR5 and CD4 antibodies were ineffective. Moreover, a CXCR4-deficient cell line, MDA-MB-468, which was resistant to Nef-induced apoptosis, became sensitive upon transfection with a CXCR4-expressing vector. This study suggests that extracellular Nef protein could contribute to the decline of CD4 counts prior to and during the onset of AIDS in patients with human immunodeficiency virus type 1 infections. PMID:14990729

  7. Relative Resistance of HLA-B to Downregulation by Naturally Occurring HIV-1 Nef Sequences.

    PubMed

    Mahiti, Macdonald; Toyoda, Mako; Jia, Xiaofei; Kuang, Xiaomei T; Mwimanzi, Francis; Mwimanzi, Philip; Walker, Bruce D; Xiong, Yong; Brumme, Zabrina L; Brockman, Mark A; Ueno, Takamasa

    2016-01-19

    HIV-1 Nef binds to the cytoplasmic region of HLA-A and HLA-B and downregulates these molecules from the surface of virus-infected cells, thus evading immune detection by CD8(+) T cells. Polymorphic residues within the HLA cytoplasmic region may affect Nef's downregulation activity. However, the effects of HLA polymorphisms on recognition by primary Nef isolates remain elusive, as do the specific Nef regions responsible for downregulation of HLA-A versus HLA-B. Here, we examined 46 Nef clones isolated from chronically HIV-1 subtype B-infected subjects for their ability to downregulate various HLA-A, HLA-B, and HLA-C molecules on the surface of virus-infected cells. Overall, HLA-B exhibited greater resistance to Nef-mediated downregulation than HLA-A, regardless of the cell type examined. As expected, no Nef clone downregulated HLA-C. Importantly, the differential abilities of patient-derived Nef clones to downregulate HLA-A and HLA-B correlated inversely with the sensitivities of HIV-infected target cells to recognition by effector cells expressing an HIV-1 Gag-specific T cell receptor. Nef codon function analysis implicated amino acid variation at position 202 (Nef-202) in differentially affecting the ability to downregulate HLA-A and HLA-B, an observation that was subsequently confirmed by experiments using Nef mutants constructed by site-directed mutagenesis. The in silico and mutagenesis analyses further suggested that Nef-202 may interact with the C-terminal Cys-Lys-Val residues of HLA-A, which are absent in HLA-B. Taken together, the results show that natural polymorphisms within Nef modulate its interaction with natural polymorphisms in the HLA cytoplasmic tails, thereby affecting the efficiency of HLA downregulation and consequent recognition by HIV-specific T cells. These results thus extend our understanding of this complex pathway of retroviral immune evasion. Recognition of genetically diverse pathogens by the adaptive immune system represents a primary

  8. HIV-1 Nef sequesters MHC-I intracellularly by targeting early stages of endocytosis and recycling

    PubMed Central

    Dirk, Brennan S.; Pawlak, Emily N.; Johnson, Aaron L.; Van Nynatten, Logan R.; Jacob, Rajesh A.; Heit, Bryan; Dikeakos, Jimmy D.

    2016-01-01

    A defining characteristic of HIV-1 infection is the ability of the virus to persist within the host. Specifically, MHC-I downregulation by the HIV-1 accessory protein Nef is of critical importance in preventing infected cells from cytotoxic T-cell mediated killing. Nef downregulates MHC-I by modulating the host membrane trafficking machinery, resulting in the endocytosis and eventual sequestration of MHC-I within the cell. In the current report, we utilized the intracellular protein-protein interaction reporter system, bimolecular fluorescence complementation (BiFC), in combination with super-resolution microscopy, to track the Nef/MHC-I interaction and determine its subcellular localization in cells. We demonstrate that this interaction occurs upon Nef binding the MHC-I cytoplasmic tail early during endocytosis in a Rab5-positive endosome. Disruption of early endosome regulation inhibited Nef-dependent MHC-I downregulation, demonstrating that Nef hijacks the early endosome to sequester MHC-I within the cell. Furthermore, super-resolution imaging identified that the Nef:MHC-I BiFC complex transits through both early and late endosomes before ultimately residing at the trans-Golgi network. Together we demonstrate the importance of the early stages of the endocytic network in the removal of MHC-I from the cell surface and its re-localization within the cell, which allows HIV-1 to optimally evade host immune responses. PMID:27841315

  9. HIV-1 evolution in patients undergoing immunotherapy with Tat, Rev, and Nef expressing dendritic cells followed by treatment interruption.

    PubMed

    de Goede, Anna L; van Deutekom, Hanneke W M; Vrancken, Bram; Schutten, Martin; Allard, Sabine D; van Baalen, Carel A; Osterhaus, Albert D M E; Thielemans, Kris; Aerts, Joeri L; Keşmir, Can; Lemey, Philippe; Gruters, Rob A

    2013-11-13

    This study aimed to evaluate HIV sequence evolution in whole genes and in CD8 T-cell epitope regions following immunotherapy and subsequent analytical treatment interruption (ATI). A second objective of this study was to analyze associations between vaccine-specific immune responses and epitope mutation rates. HIV-1-infected patients on combined antiretroviral therapy (cART) were subjected to immunotherapy by the administration of an autologous dendritic cell-based therapeutic vaccine expressing Tat, Rev, and Nef and subsequent ATI. HIV-1 genes were amplified and sequenced from plasma RNA obtained before initiation of cART as well as during ATI. Control sequences for virus evolution in untreated HIV-1-infected individuals were obtained from the HIV Sequence Database (Los Alamos). CD8 T-cell epitope regions were defined based on literature data and prediction models. HIV-1-specific immune responses were evaluated to analyze their impact on sequence evolution. Viral sequence evolution in the tat, rev, and nef genes of vaccinated patients was similar to that of controls. The number of mutations observed inside and outside CD8 T-cell epitopes was comparable for vaccine-targeted and nontargeted proteins. We found no evidence for an impact of vaccine-induced or enhanced immune responses on the number of mutations inside or outside epitopes. Therapeutic vaccination of HIV-1-infected patients with a dendritic cell-based vaccine targeting Tat, Rev, and Nef did not affect virus evolution at the whole gene level nor at the CD8 T-cell epitope level.

  10. Human immunodeficiency virus (HIV) nef-specific cytotoxic T lymphocytes in noninfected heterosexual contact of HIV-infected patients.

    PubMed Central

    Langlade-Demoyen, P; Ngo-Giang-Huong, N; Ferchal, F; Oksenhendler, E

    1994-01-01

    We report on the detection of HIV-specific cytotoxic T lymphocytes (CTL) among 23 regular partners of HIV-infected individuals. 15 of the 46 individuals enrolled in the study were positive for HLA-A2.1 typing. Among the 23 contacts studied, 7 were seropositive and 16 were seronegative on repeated tests. None of the 16 seronegative contacts were positive for p24 antigenemia nor were they positive by the lymphocytes coculture assay, although, in two instances HIV-1 DNA could be detected by PCR (in one case using a gag SK 38/39 primer, and in the other using a primer for the pol P3/P4 primer). These two individuals remained seronegative for 18 and 36 mo, respectively. HIV-specific cytotoxicity was performed in the 15 HLA-A2.1 subjects (7 indexes, 2 seropositive contacts, and 6 seronegative contacts) and in 4 HLA-matched HIV negative donors. CTL specific for env, gag, or nef proteins could not be detected in unstimulated bulk cultures of peripheral blood lymphocytes in any of the six seronegative contacts. However, using a limiting dilution assay we found an usually high frequency of HIV nef-specific CTL precursors (CTLp) for HIV env and gag was very similar to that observed in seronegative HLA-matched healthy donors. Because no presence of HIV could be demonstrated in these individuals, these findings argue against the possibility of a silent HIV infection and suggest that a CTL response against nef may be involved in a rapid and effective clearance of the virus after sexual exposure. PMID:8132769

  11. Mouse Models for Studies of In Vivo Functions of HIV-1 Nef.

    PubMed

    Zou, Wei; Zhang, Lunli

    2016-01-01

    In vitro studies have demonstrated that HIV-1 Nef has several important activities, promoting viral replication and pathogenesis. These activities include downregulation of cell surface molecules CD4 and major histocompatibility complex class I, enhancement of viral infectivity, activation of p21-activated kinase 2, and inhibition of immunoglobulin class switching. But how important each in vitro activity is to in vivo Nef function remains elusive. To address this question, several small animal models have been developed in the past two decades, such as Nef transgenic mice, SCID-hu mice, and humanized mice. Each of those models has its own pros and cons. Easy access and relative inexpensiveness have made small animal models the favorite models for HIV research. This review will be focused on the recent progress in the understanding of the in vivo functions of HIV-1 Nef obtained from studies using these small animal models.

  12. Evidence for Adaptive Evolution at the Divergence Between Lymphoid and Brain HIV-1 nef Genes

    PubMed Central

    Olivieri, Kevin C.; Agopian, Kristin A.; Mukerji, Joya

    2010-01-01

    Abstract Human immunodeficiency virus type 1 (HIV) infection of the central nervous system frequently causes HIV-associated neurocognitive disorders (HAND). The role of HIV Nef and other accessory proteins in HAND pathogenesis is unclear. To determine whether HIV nef undergoes adaptive selection in brain, we cloned 100 nef sequences (n = 30 brain and n = 70 lymphoid) from four patients with AIDS and HIV-associated dementia (HAD). Normalized nonsynonymous substitutions were more frequent at the divergence of lymphoid and brain sequences, indicating stronger adaptive selection in brain compared to lymphoid tissue. Brain-specific nonsynonymous substitutions were found within an NH3-terminal CTL epitope, the PACS1 binding motif, or positions predicted to be important for activation of the myeloid-restricted Src family tyrosine kinase Hck. These results suggest that adaptive selection of HIV nef in brain may reflect altered requirements for efficient replication in macrophages and brain-specific immune selection pressures. PMID:20377428

  13. p21-Activated Kinase 1 Plays a Critical Role in Cellular Activation by Nef

    PubMed Central

    Fackler, Oliver T.; Lu, Xiaobin; Frost, Jeffrey A.; Geyer, Matthias; Jiang, Bing; Luo, Wen; Abo, Arie; Alberts, Arthur S.; Peterlin, B. Matija

    2000-01-01

    The activation of Nef-associated kinase (NAK) by Nef from human and simian immunodeficiency viruses is critical for efficient viral replication and pathogenesis. This induction occurs via the guanine nucleotide exchange factor Vav and the small GTPases Rac1 and Cdc42. In this study, we identified NAK as p21-activated kinase 1 (PAK1). PAK1 bound to Nef in vitro and in vivo. Moreover, the induction of cytoskeletal rearrangements such as the formation of trichopodia, the activation of Jun N-terminal kinase, and the increase of viral production were blocked by an inhibitory peptide that targets the kinase activity of PAK1 (PAK1 83-149). These results identify NAK as PAK1 and emphasize the central role its kinase activity plays in cytoskeletal rearrangements and cellular signaling by Nef. PMID:10713183

  14. HIV's Nef interacts with β-catenin of the Wnt signaling pathway in HEK293 cells.

    PubMed

    Weiser, Keren; Barton, Meredith; Gershoony, Dafna; Dasgupta, Ramanuj; Cardozo, Timothy

    2013-01-01

    The Wnt signaling pathway is implicated in major physiologic cellular functions, such as proliferation, migration, cell fate specification, maintenance of pluripotency and induction of tumorigenicity. Proliferation and migration are important responses of T-cells, which are major cellular targets of HIV infection. Using an informatics screen, we identified a previously unsuspected interaction between HIV's Nef protein and β-catenin, a key component of the Wnt pathway. A segment in Nef contains identical amino acids at key positions and structurally mimics the β-catenin binding sites on endogenous β-catenin ligands. The interaction between Nef and β-catenin was confirmed in vitro and in a co-immunoprecipitation from HEK293 cells. Moreover, the introduction of Nef into HEK293 cells specifically inhibited a Wnt pathway reporter.

  15. High pressure liquid level monitor

    DOEpatents

    Bean, Vern E.; Long, Frederick G.

    1984-01-01

    A liquid level monitor for tracking the level of a coal slurry in a high-pressure vessel including a toroidal-shaped float with magnetically permeable bands thereon disposed within the vessel, two pairs of magnetic field generators and detectors disposed outside the vessel adjacent the top and bottom thereof and magnetically coupled to the magnetically permeable bands on the float, and signal processing circuitry for combining signals from the top and bottom detectors for generating a monotonically increasing analog control signal which is a function of liquid level. The control signal may be utilized to operate high-pressure control valves associated with processes in which the high-pressure vessel is used.

  16. Human Immunodeficiency Virus nef signature sequences are associated with pulmonary hypertension.

    PubMed

    Almodovar, Sharilyn; Knight, Rob; Allshouse, Amanda A; Roemer, Sarah; Lozupone, Catherine; McDonald, Daniel; Widmann, Jeremy; Voelkel, Norbert F; Shelton, Robert J; Suarez, Edu B; Hammer, Kenneth W; Goujard, Cecile; Petrosillo, Nicola; Simonneau, Gerald; Hsue, Priscilla Y; Humbert, Marc; Flores, Sonia C

    2012-06-01

    Severe pulmonary hypertension (PH) associated with vascular remodeling is a long-term complication of HIV infection (HIV-PH) affecting 1/200 infected individuals vs. 1/200,000 frequency in the uninfected population. Factors accounting for increased PH susceptibility in HIV-infected individuals are unknown. Rhesus macaques infected with chimeric SHIVnef virions but not with SIV display PH-like pulmonary vascular remodeling suggesting that HIV-Nef is associated with PH; these monkeys showed changes in nef sequences that correlated with pathogenesis after passage in vivo. We further examined whether HIV-nef alleles in HIV-PH subjects have signature sequences associated with the disease phenotype. We evaluated specimens from participants with and without HIV-PH from European Registries and validated results with samples collected as part of the Lung-HIV Studies in San Francisco. We found that 10 polymorphisms in nef were overrepresented in blood cells or lung tissue specimens from European HIV-PH individuals but significantly less frequent in HIV-infected individuals without PH. These polymorphisms mapped to known functional domains in Nef. In the validation cohort, 7/10 polymorphisms in the HIV-nef gene were confirmed; these polymorphisms arose independently from viral load, CD4(+) T cell counts, length of infection, and antiretroviral therapy status. Two out of 10 polymorphisms were previously reported in macaques with PH-like pulmonary vascular remodeling. Cloned recombinant Nef proteins from clinical samples down-regulated CD4, suggesting that these primary isolates are functional. This study offers new insights into the association between Nef polymorphisms in functional domains and the HIV-PH phenotype. The utility of these polymorphisms as predictors of PH should be examined in a larger population.

  17. High-Level Radioactive Waste.

    ERIC Educational Resources Information Center

    Hayden, Howard C.

    1995-01-01

    Presents a method to calculate the amount of high-level radioactive waste by taking into consideration the following factors: the fission process that yields the waste, identification of the waste, the energy required to run a 1-GWe plant for one year, and the uranium mass required to produce that energy. Briefly discusses waste disposal and…

  18. High-Level Radioactive Waste.

    ERIC Educational Resources Information Center

    Hayden, Howard C.

    1995-01-01

    Presents a method to calculate the amount of high-level radioactive waste by taking into consideration the following factors: the fission process that yields the waste, identification of the waste, the energy required to run a 1-GWe plant for one year, and the uranium mass required to produce that energy. Briefly discusses waste disposal and…

  19. Human Immunodeficiency Virus Type 1 Nef-Induced CD4 Cell Surface Downregulation Is Inhibited by Ikarugamycin

    PubMed Central

    Luo, Tianci; Fredericksen, Brenda L.; Hasumi, Keiji; Endo, Akira; Garcia, J. Victor

    2001-01-01

    One well-characterized in vitro function of Nef is its ability to remove CD4, the human immunodeficiency virus (HIV) receptor, from the cell surface. Nef accomplishes this by accelerating the internalization and degradation of CD4. Current models propose that Nef promotes CD4 internalization via an increased association of CD4 with clathrin-coated pits (CCP). Here, we investigated the effect of a naturally occurring antiprotozoan antibiotic, ikarugamycin (IKA), on CD4 cell surface expression in human monocytic cells stably expressing HIV type 1 SF2 Nef. IKA was able to efficiently restore CD4 cell surface expression in Nef-expressing cells without affecting either CD4 synthesis or Nef expression. In addition, we demonstrate that IKA is also capable of efficiently blocking CD4 down-modulation in response to phorbol myristate acetate. Our data suggest that IKA may be an efficient and useful inhibitor of CCP-dependent endocytosis. PMID:11160755

  20. The CMS High Level Trigger

    NASA Astrophysics Data System (ADS)

    Trocino, Daniele

    2014-06-01

    The CMS experiment has been designed with a two-level trigger system: the Level-1 Trigger, implemented in custom-designed electronics, and the High-Level Trigger (HLT), a streamlined version of the CMS offline reconstruction software running on a computer farm. A software trigger system requires a tradeoff between the complexity of the algorithms running with the available computing power, the sustainable output rate, and the selection efficiency. We present the performance of the main triggers used during the 2012 data taking, ranging from simple single-object selections to more complex algorithms combining different objects, and applying analysis-level reconstruction and selection. We discuss the optimisation of the trigger and the specific techniques to cope with the increasing LHC pile-up, reducing its impact on the physics performance.

  1. A basic patch on alpha-adaptin is required for binding of human immunodeficiency virus type 1 Nef and cooperative assembly of a CD4-Nef-AP-2 complex.

    PubMed

    Chaudhuri, Rittik; Mattera, Rafael; Lindwasser, O Wolf; Robinson, Margaret S; Bonifacino, Juan S

    2009-03-01

    A critical function of the human immunodeficiency virus type 1 Nef protein is the downregulation of CD4 from the surfaces of infected cells. Nef is believed to act by linking the cytosolic tail of CD4 to the endocytic machinery, thereby increasing the rate of CD4 internalization. In support of this model, weak binary interactions between CD4, Nef, and the endocytic adaptor complex, AP-2, have been reported. In particular, dileucine and diacidic motifs in the C-terminal flexible loop of Nef have been shown to mediate binding to a combination of the alpha and sigma2 subunits of AP-2. Here, we report the identification of a potential binding site for the Nef diacidic motif on alpha-adaptin. This site comprises two basic residues, lysine-297 and arginine-340, on the alpha-adaptin trunk domain. The mutation of these residues specifically inhibits the ability of Nef to bind AP-2 and downregulate CD4. We also present evidence that the diacidic motif on Nef and the basic patch on alpha-adaptin are both required for the cooperative assembly of a CD4-Nef-AP-2 complex. This cooperativity explains how Nef is able to efficiently downregulate CD4 despite weak binary interactions between components of the tripartite complex.

  2. RPython high-level synthesis

    NASA Astrophysics Data System (ADS)

    Cieszewski, Radoslaw; Linczuk, Maciej

    2016-09-01

    The development of FPGA technology and the increasing complexity of applications in recent decades have forced compilers to move to higher abstraction levels. Compilers interprets an algorithmic description of a desired behavior written in High-Level Languages (HLLs) and translate it to Hardware Description Languages (HDLs). This paper presents a RPython based High-Level synthesis (HLS) compiler. The compiler get the configuration parameters and map RPython program to VHDL. Then, VHDL code can be used to program FPGA chips. In comparison of other technologies usage, FPGAs have the potential to achieve far greater performance than software as a result of omitting the fetch-decode-execute operations of General Purpose Processors (GPUs), and introduce more parallel computation. This can be exploited by utilizing many resources at the same time. Creating parallel algorithms computed with FPGAs in pure HDL is difficult and time consuming. Implementation time can be greatly reduced with High-Level Synthesis compiler. This article describes design methodologies and tools, implementation and first results of created VHDL backend for RPython compiler.

  3. Discovery of a diaminoquinoxaline benzenesulfonamide antagonist of HIV-1 Nef function using a yeast-based phenotypic screen

    PubMed Central

    2013-01-01

    Background HIV-1 Nef is a viral accessory protein critical for AIDS progression. Nef lacks intrinsic catalytic activity and binds multiple host cell signaling proteins, including Hck and other Src-family tyrosine kinases. Nef binding induces constitutive Hck activation that may contribute to HIV pathogenesis by promoting viral infectivity, replication and downregulation of cell-surface MHC-I molecules. In this study, we developed a yeast-based phenotypic screen to identify small molecules that inhibit the Nef-Hck complex. Results Nef-Hck interaction was faithfully reconstituted in yeast cells, resulting in kinase activation and growth arrest. Yeast cells expressing the Nef-Hck complex were used to screen a library of small heterocyclic compounds for their ability to rescue growth inhibition. The screen identified a dihydrobenzo-1,4-dioxin-substituted analog of 2-quinoxalinyl-3-aminobenzene-sulfonamide (DQBS) as a potent inhibitor of Nef-dependent HIV-1 replication and MHC-I downregulation in T-cells. Docking studies predicted direct binding of DQBS to Nef which was confirmed in differential scanning fluorimetry assays with recombinant purified Nef protein. DQBS also potently inhibited the replication of HIV-1 NL4-3 chimeras expressing Nef alleles representative of all M-group HIV-1 clades. Conclusions Our findings demonstrate the utility of a yeast-based growth reversion assay for the identification of small molecule Nef antagonists. Inhibitors of Nef function discovered with this assay, such as DQBS, may complement the activity of current antiretroviral therapies by enabling immune recognition of HIV-infected cells through the rescue of cell surface MHC-I. PMID:24229420

  4. Validation of a novel secretion modification region (SMR) of HIV-1 Nef using cohort sequence analysis and molecular modeling

    PubMed Central

    Campbell, Patrick E.; Isayev, Olexandr; Ali, Syed A.; Roth, William W.; Huang, Ming-Bo; Powell, Michael D.; Leszczynski, Jerzy

    2012-01-01

    The HIV-1 accessory protein Nef plays an active role in the pathogenesis of AIDS by its numerous cellular interactions that facilitate the release of virus particles. This 27 kDa protein is required for maintenance of the viral replication in HIV, and is also known to contribute to immune evasion, blocking of apoptosis in virus-infected cells and enhancement of virus infectivity. Nef has been shown to be secreted and is present on the surface of virus-infected cells. Recent studies from our laboratory have shown that the Nef protein is secreted from nef-transfected and HIV-1-infected cells in small exosome-like vesicles (40–100 nm diam.) that do not contain virions. We have identified three amino-terminal domains of Nef as necessary for secretion: (i) the four arginine residues (17,19,21, 22) comprising the basic region; (ii) the phosphofurin acidic cluster sequence (PACS) composed of four glutamic acid residues (61–64); (iii) a previously unknown motif spanning amino acid residues 65–69 (VGFPV) which we named the secretion modification region (SMR). In this study, we have used population-based phylogeny data and sequence analysis to characterize the conservation of the Nef SMR domain that regulates vesicle secretion. We have performed in silico computational chemistry analysis involving molecular dynamic structure modeling of mutations in the SMR motif. Sequence analysis of Nef from HIV-1-infected patients, including slow progressors (SP), long term progressors (LTP) and long term non-progressors (LTNP) demonstrated 99 % conservation of the Nef SMR motif. Computational analysis including modeling of wild-type HIV-1 Nef and V66A Nef SMR mutant using structural homology and molecular dynamics of ligand-associated interactions indicated significant structural changes in the Nef mutant, thus supporting the importance of the SMR domain for mediating Nef vesicle secretion. PMID:22643973

  5. CD4 down-modulation during infection of human T cells with human immunodeficiency virus type 1 involves independent activities of vpu, env, and nef.

    PubMed Central

    Chen, B K; Gandhi, R T; Baltimore, D

    1996-01-01

    The human immunodeficiency virus type 1 (HIV-1) genes vpu, env, and nef have all been implicated in modulating the levels of cell surface CD4 on infected cells. To quantitatively assess the relative contribution of each gene product to the regulation of CD4 during HIV infection of Jurkat T cells and peripheral blood mononuclear cells, we have developed an infectious HIV reporter system which expresses different combinations of these genes. To distinguish infected cells in the early or late stages of infection from uninfected cells, these viruses were designed to express human placental alkaline phosphatase with the kinetics of either early or late viral genes. Flow cytometry to detect placental alkaline phosphatase and CD4 in infected cells showed that vpu, env, and nef are independently capable of down-modulation of CD4. As predicted by their respective expression patterns, nef down-modulated CD4 rapidly during the early phase of virus infection whereas vpu and env functioned late in the infection. In both Jurkat cells and peripheral blood mononuclear cells, a combination of the three genes was more efficient than any one or two genes, demonstrating that all three genes are required to achieve maximal CD4 down-modulation. In primary cells, down-modulation of CD4 was less efficient than in Jurkat cells and there was a stronger dependence on nef function for reducing cell surface CD4. HIV therefore has three genes that are able to independently down-modulate CD4; together, they can eliminate the bulk of cell surface CD4. PMID:8709227

  6. HIV-Nef and ADAM17-Containing Plasma Extracellular Vesicles Induce and Correlate with Immune Pathogenesis in Chronic HIV Infection

    PubMed Central

    Lee, Jung-Hyun; Schierer, Stephan; Blume, Katja; Dindorf, Jochen; Wittki, Sebastian; Xiang, Wei; Ostalecki, Christian; Koliha, Nina; Wild, Stefan; Schuler, Gerold; Fackler, Oliver T.; Saksela, Kalle; Harrer, Thomas; Baur, Andreas S.

    2016-01-01

    Antiretroviral therapy (ART) efficiently suppresses HIV replication but immune activation and low CD4 T cell counts often persist. The underlying mechanism of this ART-resistant pathogenesis is not clear. We observed that levels of plasma extracellular vesicles (pEV) are strongly elevated in HIV infection and do not decline during ART. Surprisingly, these vesicles contained the viral accessory proteins Nef and Vpu, which are assumed to be not expressed under efficient ART, as well as pro-inflammatory effectors, including activated ADAM17. HIV pEV were characterized by the presence of activated αvβ3 and absence of CD81 and Tsg101. Correlating with immune activation, peripheral monocytes ingested large amounts of pEV, giving rise to an increased population of CD1c+ CD14+ cells that secreted inflammatory cytokines. Importantly, the pro-inflammatory content, particularly ADAM17 activity, correlated with low T cell counts. Preliminary evidence suggested that HIV pEV derived from peripheral mononuclear cells and from an unknown myeloid cell population. In summary we propose an important role of pro-inflammatory pEV in chronic HIV infection due to ongoing viral Nef activity. PMID:27211553

  7. HIV-Nef and ADAM17-Containing Plasma Extracellular Vesicles Induce and Correlate with Immune Pathogenesis in Chronic HIV Infection.

    PubMed

    Lee, Jung-Hyun; Schierer, Stephan; Blume, Katja; Dindorf, Jochen; Wittki, Sebastian; Xiang, Wei; Ostalecki, Christian; Koliha, Nina; Wild, Stefan; Schuler, Gerold; Fackler, Oliver T; Saksela, Kalle; Harrer, Thomas; Baur, Andreas S

    2016-04-01

    Antiretroviral therapy (ART) efficiently suppresses HIV replication but immune activation and low CD4 T cell counts often persist. The underlying mechanism of this ART-resistant pathogenesis is not clear. We observed that levels of plasma extracellular vesicles (pEV) are strongly elevated in HIV infection and do not decline during ART. Surprisingly, these vesicles contained the viral accessory proteins Nef and Vpu, which are assumed to be not expressed under efficient ART, as well as pro-inflammatory effectors, including activated ADAM17. HIV pEV were characterized by the presence of activated αvβ3 and absence of CD81 and Tsg101. Correlating with immune activation, peripheral monocytes ingested large amounts of pEV, giving rise to an increased population of CD1c(+) CD14(+) cells that secreted inflammatory cytokines. Importantly, the pro-inflammatory content, particularly ADAM17 activity, correlated with low T cell counts. Preliminary evidence suggested that HIV pEV derived from peripheral mononuclear cells and from an unknown myeloid cell population. In summary we propose an important role of pro-inflammatory pEV in chronic HIV infection due to ongoing viral Nef activity.

  8. The Potency of Nef-Mediated SERINC5 Antagonism Correlates with the Prevalence of Primate Lentiviruses in the Wild.

    PubMed

    Heigele, Anke; Kmiec, Dorota; Regensburger, Kerstin; Langer, Simon; Peiffer, Lukas; Stürzel, Christina M; Sauter, Daniel; Peeters, Martine; Pizzato, Massimo; Learn, Gerald H; Hahn, Beatrice H; Kirchhoff, Frank

    2016-09-14

    The cellular factor serine incorporator 5 (SERINC5) impairs HIV-1 infectivity but is antagonized by the viral Nef protein. We analyzed the anti-SERINC5 activity of Nef proteins across primate lentiviruses and examined whether SERINC5 represents a barrier to cross-species transmissions and/or within-species viral spread. HIV-1, HIV-2, and SIV Nefs counteract human, ape, monkey, and murine SERINC5 orthologs with similar potency. However, HIV-1 Nefs are more active against SERINC5 than HIV-2 Nefs, and chimpanzee SIV (SIVcpz) Nefs are more potent than those of their monkey precursors. Additionally, Nefs of HIV and most SIVs rely on the dileucine motif in the C-terminal loop for anti-SERINC5 activity, while the Nef from colobus SIV (SIVcol) evolved different inhibitory mechanisms. We also found a significant correlation between anti-SERINC5 potency and the SIV prevalence in the respective ape and monkey species. Thus, Nef-mediated SERINC5 antagonism may determine the ability of primate lentiviruses to spread within natural hosts. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. SIMULTANEOUS ASSESSMENT OF CD4 AND MHCI DOWNREGULATION BY NEF PRIMARY ISOLATES IN THE CONTEXT OF INFECTION

    PubMed Central

    Ali, Ayub; Realegeno, Susan; Yang, Otto O.; Lewis, Martha J.

    2009-01-01

    SUMMARY The HIV-1 Nef protein plays a key role in pathogenesis, as demonstrated by strong selective pressure to maintain its open reading frame, and disease attenuation when it is deleted. Among myriad cellular effects attributed to Nef, downregulation of cell surface CD4 and major histocompatibility complex class I (MHC-I) proteins are the best documented. However, few data regarding primary isolate Nef functions are available, and most studies have been performed using transient transfections to express Nef driven by a non-physiologic promoter. A novel assay system to measure simultaneously the downregulation of CD4 and MHC-I by primary HIV-1 nef in a more physiologic viral genomic context is presented. Examination of plasma nef mixtures allowed comprehensive profiling of these Nef functions within the quasispecies in vivo. Subsets within the circulating nef population were observed that are either fully functional or non-functional. These data demonstrated that this assay system allows rapid characterization of bulk and clonal Nef functional profiles that can be used in pathogenesis studies to define further its important role in pathogenesis. PMID:19643141

  10. The ALICE high level trigger

    NASA Astrophysics Data System (ADS)

    Alt, T.; Grastveit, G.; Helstrup, H.; Lindenstruth, V.; Loizides, C.; Röhrich, D.; Skaali, B.; Steinbeck, T.; Stock, R.; Tilsner, H.; Ullaland, K.; Vestbø, A.; Vik, T.; Wiebalck, A.; the ALICE Collaboration

    2004-08-01

    The ALICE experiment at LHC will implement a high-level trigger system for online event selection and/or data compression. The largest computing challenge is posed by the TPC detector, which requires real-time pattern recognition. The system entails a very large processing farm that is designed for an anticipated input data stream of 25 GB s-1. In this paper, we present the architecture of the system and the current state of the tracking methods and data compression applications.

  11. High-Level Connectionist Models

    DTIC Science & Technology

    1993-10-01

    Artficial Intelligence Research Computer and Information Science Department The Ohio State Universiy Columbus, Ohio 43210 pja@ci.ohio-state.edu saunders...Peter J. Angeline, Gregory M. Saunders and Jordan B. Pollack Laboratory for Artficial Intelligence Research Computer and 1i4ormadon Science Deparment...AD-A273 638 OHIOi High-Level Connectionist Models 5LPJE UNIVERSITY Jordan B. Pollack Laboratory for Artificial Intelligence Research Department of

  12. Trans-Cellular Introduction of HIV-1 Protein Nef Induces Pathogenic Response in Caenorhabditis elegans

    PubMed Central

    Nazir, Aamir; Sammi, Shreesh Raj; Singh, Pankaj; Tripathi, Raj Kamal

    2010-01-01

    Background Caenorhabditis elegans has emerged as a very powerful model for studying the host pathogen interactions. Despite the absence of a naturally occurring viral infection for C. elegans, the model is now being exploited experimentally to study the basic aspects of virus-host interplay. The data generated from recent studies suggests that the virus that infects mammalian cells does infect, replicate and accumulate in C. elegans. Methodology/Principal Findings We took advantage of the easy-to-achieve protein introduction in C. elegans and employing the methodology, we administered HIV-1 protein Nef into live worms. Nef is known to be an important protein for exacerbating HIV-1 pathogenesis in host by enhancing viral replication. The deletion of nef from the viral genome has been reported to inhibit its replication in the host, thereby leading to delayed pathogenesis. Our studies, employing Nef introduction into C. elegans, led to creation of an in-vivo model that allowed us to study, whether or not, the protein induces effect in the whole organism. We observed a marked lipodystrophy, effect on neuromuscular function, impaired fertility and reduced longevity in the worms exposed to Nef. The observed effects resemble to those observed in Nef transgenic mice and most interestingly the effects also relate to some of the pathogenic aspects exhibited by human AIDS patients. Conclusions/Significance Our studies underline the importance of this in vivo model for studying the interactions of Nef with host proteins, which could further be used for identifying possible inhibitors of such interactions. PMID:21179446

  13. Interaction between HIV-1 Nef and calnexin: from modeling to small molecule inhibitors reversing HIV-induced lipid accumulation

    PubMed Central

    Hunegnaw, Ruth; Vassylyeva, Marina; Dubrovsky, Larisa; Pushkarsky, Tatiana; Sviridov, Dmitri; Anashkina, Anastasia A.; Üren, Aykut; Brichacek, Beda; Vassylyev, Dmitry; Adzhubei, Alexei A.; Bukrinsky, Michael

    2016-01-01

    Objective HIV-infected patients are at an increased risk of developing atherosclerosis, in part due to downmodulation and functional impairment of ATP-Binding Cassette A1 (ABCA1) cholesterol transporter by the HIV-1 protein Nef. The mechanism of this effect involves Nef interacting with an endoplasmic reticulum (ER) chaperone calnexin and disrupting calnexin binding to ABCA1, leading to ABCA1 retention in ER, its degradation and resulting suppression of cholesterol efflux. However, molecular details of Nef-calnexin interaction remained unknown, limiting translational impact of this finding. Approach and results Here, we used molecular modeling and mutagenesis to characterize Nef-calnexin interaction and to identify small molecule compounds that could block it. We demonstrated that interaction between Nef and calnexin is direct and can be reconstituted using recombinant proteins in vitro with a binding affinity of 89.1 nM measured by surface plasmon resonance. The cytoplasmic tail of calnexin is essential and sufficient for interaction with Nef, and binds Nef with affinity of 9.4 nM. Replacing lysine residues in positions 4 and 7 of Nef with alanines abrogates Nef-calnexin interaction, prevents ABCA1 downregulation by Nef, and preserves cholesterol efflux from HIV-infected cells. Through virtual screening of the NCI library of compounds, we identified a compound, 1[(7-Oxo-7H-benz[de]anthracene-3-yl)amino]anthraquinone, which blocked Nef-calnexin interaction, partially restored ABCA1 activity in HIV-infected cells, and reduced foam cell formation in a culture of HIV-infected macrophages. Conclusion This study identifies potential targets that can be exploited to block the pathogenic effect of HIV infection on cholesterol metabolism and prevent atherosclerosis in HIV-infected subjects. PMID:27470515

  14. Nef proteins of epidemic HIV-1 group O strains antagonize human tetherin.

    PubMed

    Kluge, Silvia F; Mack, Katharina; Iyer, Shilpa S; Pujol, François M; Heigele, Anke; Learn, Gerald H; Usmani, Shariq M; Sauter, Daniel; Joas, Simone; Hotter, Dominik; Bibollet-Ruche, Frederic; Plenderleith, Lindsey J; Peeters, Martine; Geyer, Matthias; Sharp, Paul M; Fackler, Oliver T; Hahn, Beatrice H; Kirchhoff, Frank

    2014-11-12

    Most simian immunodeficiency viruses use their Nef protein to antagonize the host restriction factor tetherin. A deletion in human tetherin confers Nef resistance, representing a hurdle to successful zoonotic transmission. HIV-1 group M evolved to utilize the viral protein U (Vpu) to counteract tetherin. Although HIV-1 group O has spread epidemically in humans, it has not evolved a Vpu-based tetherin antagonism. Here we show that HIV-1 group O Nef targets a region adjacent to this deletion to inhibit transport of human tetherin to the cell surface, enhances virion release, and increases viral resistance to inhibition by interferon-α. The Nef protein of the inferred common ancestor of group O viruses is also active against human tetherin. Thus, Nef-mediated antagonism of human tetherin evolved prior to the spread of HIV-1 group O and likely facilitated secondary virus transmission. Our results may explain the epidemic spread of HIV-1 group O. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. IL-12 and GM-CSF in DNA/MVA Immunizations against HIV-1 CRF12_BF Nef Induced T-Cell Responses With an Enhanced Magnitude, Breadth and Quality

    PubMed Central

    Rodríguez, Ana María; Falivene, Juliana; Holgado, María Pía; Turk, Gabriela; Gherardi, María Magdalena

    2012-01-01

    In Argentina, the HIV epidemic is characterized by the co-circulation of subtype B and BF recombinant viral variants. Nef is an HIV protein highly variable among subtypes, making it a good tool to study the impact of HIV variability in the vaccine design setting. We have previously reported a specific cellular response against NefBF with low cross-reactivity to NefB in mice. The aim of this work was to analyze whether the co-administration of IL-12 and GM-CSF, using DNA and MVA vaccine vectors, could improve the final cellular response induced. Mice received three DNA priming doses of a plasmid that express NefBF plus DNAs expressing IL-12 and/or GM-CSF. Afterwards, all the groups were boosted with a MVAnefBF dose. The highest increase in the magnitude of the NefBF response, compared to that induced in the control was found in the IL-12 group. Importantly, a response with higher breadth was detected in groups which received IL-12 or GM-CSF, evidenced as an increased frequency of recognition of homologous (BF) and heterologous (B) Nef peptides, as well as a higher number of other Nef peptide pools representing different viral subtypes. However, these improvements were lost when both DNA cytokines were simultaneously administered, as the response was focused against the immunodominant peptide with a detrimental response towards subdominant epitopes. The pattern of cytokines secreted and the specific-T-cell proliferative capacity were improved in IL-12 and IL-12+GM-CSF groups. Importantly IL-12 generated a significant higher T-cell avidity against a B heterologous peptide. This study indicates that the incorporation of DNA expressing IL-12 in DNA/MVA schemes produced the best results in terms of improvements of T-cell-response key properties such as breadth, cross-reactivity and quality (avidity and pattern of cytokines secreted). These relevant results contribute to the design of strategies aimed to induce T-cell responses against HIV antigens with higher quality

  16. HIV-1 Nef activates STAT1 in human monocytes/macrophages through the release of soluble factors.

    PubMed

    Federico, M; Percario, Z; Olivetta, E; Fiorucci, G; Muratori, C; Micheli, A; Romeo, G; Affabris, E

    2001-11-01

    Monocytes/macrophages play a predominant role in the immunologic network by secreting and reacting to a wide range of soluble factors. Human immunodeficiency virus (HIV) infection leads to deep immunologic dysfunctions, also as a consequence of alterations in the pattern of cytokine release. Recent studies on in vivo models demonstrated that the expression of HIV Nef alone mimics many pathogenetic effects of HIV infection. In particular, Nef expression in monocytes/macrophages has been correlated with remarkable modifications in the pattern of secreted soluble factors, suggesting that the interaction of Nef with monocytes/macrophages plays a role in the pathogenesis of acquired immunodeficiency syndrome (AIDS). This study sought to define possible alterations in intracellular signaling induced by Nef in monocytes/macrophages. Results demonstrate that HIV-1 Nef specifically activates both alpha and beta isoforms of the signal transducer and activator of transcription 1 (STAT1). This was observed both by infecting human monocyte-derived macrophages (MDMs) with HIV-1 deletion mutants, and by exploiting the ability of MDMs to internalize soluble, recombinant Nef protein (rNef). STAT1-alpha activation occurs on phosphorylation of both C-terminal Tyr701 and Ser727 and leads to a strong binding activity. Nef-dependent STAT1 activation is followed by increased expression of both STAT1 and interferon regulatory factor-1, a transcription factor transcriptionally regulated by STAT1 activation. It was also established that Nef-induced STAT1- alpha/beta activation occurs through the secretion of soluble factors. Taken together, the results indicate that HIV-1 Nef could interfere with STAT1-governed intracellular signaling in human monocytes/macrophages.

  17. Species-Specific Activity of SIV Nef and HIV-1 Vpu in Overcoming Restriction by Tetherin/BST2

    PubMed Central

    Neidermyer, William; Rahmberg, Andrew; Mackey, John; Fofana, Ismael Ben; Johnson, Welkin E.; Westmoreland, Susan; Evans, David T.

    2009-01-01

    Tetherin, also known as BST2, CD317 or HM1.24, was recently identified as an interferon-inducible host–cell factor that interferes with the detachment of virus particles from infected cells. HIV-1 overcomes this restriction by expressing an accessory protein, Vpu, which counteracts tetherin. Since lentiviruses of the SIVsmm/mac/HIV-2 lineage do not have a vpu gene, this activity has likely been assumed by other viral gene products. We found that deletion of the SIVmac239 nef gene significantly impaired virus release in cells expressing rhesus macaque tetherin. Virus release could be restored by expressing Nef in trans. However, Nef was unable to facilitate virus release in the presence of human tetherin. Conversely, Vpu enhanced virus release in the presence of human tetherin, but not in the presence of rhesus tetherin. In accordance with the species-specificity of Nef in mediating virus release, SIV Nef downregulated cell-surface expression of rhesus tetherin, but did not downregulate human tetherin. The specificity of SIV Nef for rhesus tetherin mapped to four amino acids in the cytoplasmic domain of the molecule that are missing from human tetherin, whereas the specificity of Vpu for human tetherin mapped to amino acid differences in the transmembrane domain. Nef alleles of SIVsmm, HIV-2 and HIV-1 were also able to rescue virus release in the presence of both rhesus macaque and sooty mangabey tetherin, but were generally ineffective against human tetherin. Thus, the ability of Nef to antagonize tetherin from these Old World primates appears to be conserved among the primate lentiviruses. These results identify Nef as the viral gene product of SIV that opposes restriction by tetherin in rhesus macaques and sooty mangabeys, and reveal species-specificity in the activities of both Nef and Vpu in overcoming tetherin in their respective hosts. PMID:19436700

  18. High level white noise generator

    DOEpatents

    Borkowski, Casimer J.; Blalock, Theron V.

    1979-01-01

    A wide band, stable, random noise source with a high and well-defined output power spectral density is provided which may be used for accurate calibration of Johnson Noise Power Thermometers (JNPT) and other applications requiring a stable, wide band, well-defined noise power spectral density. The noise source is based on the fact that the open-circuit thermal noise voltage of a feedback resistor, connecting the output to the input of a special inverting amplifier, is available at the amplifier output from an equivalent low output impedance caused by the feedback mechanism. The noise power spectral density level at the noise source output is equivalent to the density of the open-circuit thermal noise or a 100 ohm resistor at a temperature of approximately 64,000 Kelvins. The noise source has an output power spectral density that is flat to within 0.1% (0.0043 db) in the frequency range of from 1 KHz to 100 KHz which brackets typical passbands of the signal-processing channels of JNPT's. Two embodiments, one of higher accuracy that is suitable for use as a standards instrument and another that is particularly adapted for ambient temperature operation, are illustrated in this application.

  19. Potential impact of viral load and genetic makeup of HIV type 1 on mother-to-child transmission: characterization of env-C2V3C3 and nef sequences.

    PubMed

    Pádua, Elizabeth; Parreira, Ricardo; Tendeiro, Rita; Nunes, Baltazar; Castela, João; Soares, Isabel; Mouzinho, Ana; Reis, Eduarda; Paixão, Maria Teresa

    2009-11-01

    HIV-1 mother-to-child transmission (MTCT) was evaluated in terms of the molecular characterization of the env and nef genomic regions and quantification of maternal RNA viral loads. Assignment of viral subtype was achieved by direct sequencing of PCR 1172 products amplified from proviral DNA in 45 HIV-1-nontransmitting mothers (NTM), along with 13 pairs of HIV-1-transmitting mothers (TM) and their infected children (C). Analysis of the env C2V3C3 and nef sequences revealed that subtypes G and B, and their genetic combinations (AG, BG), accounted for over 84.5% of all viruses identified. The genetic structure form envA-nefG was the most commonly observed, with a lower frequency in the NTM (13.3%) compared to the TM (23.1%) group. A greater number of genetic forms was observed among NTM, namely the presence of sequences assigned to subtypes D and F, as well as the intergenetic A/J, and C/U, recombinant forms, along with a mosaic provirus with a complex putative envA-nefEGE genetic structure. No significant differences were found when RNA viral loads were evaluated as a function of the viral subtypes. Nevertheless, a relatively high quantification of HIV-1 RNA was obtained in the NTM group, emphasizing the importance of the compliance and effectiveness of therapeutic schemes to control viral replication and reduce the risk of HIV vertical transmission. V3 sequences displaying features associated with the R5 phenotype dominated in both groups. Both C2V3C3 and Nef's functional domains were conserved during HIV-1 vertical transmission.

  20. SIVsm Tat, Rev, and Nef1: functional characteristics of r-GV internalization on isotypes, cytokines, and intracellular degradation

    PubMed Central

    2010-01-01

    Background Recombinant gas vesicles (r-GV) from Halobacterium sp. strain SD109 expressing cassettes with different SIVsm inserts, have potential utility as an effective antigen display system for immunogen testing in vivo and for initial epitope assessments in vitro. Previous mouse model studies demonstrated immunization with r-GV expressing selected exogenous sequences elicited a prolonged immune response. Here we tested segments from three SIVsm genes (tat, rev, and nef) each surface displayed by r-GV. As with HIV, for SIVsm the proteins encoded by tat, rev and nef respectively serve critical and diverse functions: effects on efficient viral RNA polymerase II transcription, regulation of viral gene expression and effects on specific signaling functions through the assembly of multiprotein complexes. Humoral responses to r-GVTat, Rev or Nef1 elicited in vivo, associated changes in selected cell cytokine production following r-GV internalization, and the capacity of J774A.1 macrophage cells to degrade these internalized display/delivery particles in vitro were examined. Results The in vivo studies involving r-GV immunizations and in vitro studies of r-GV uptake by J774A.1 macrophages demonstrated: (i) tests for antibody isotypes in immunized mice sera showed activation and re-stimulation of memory B cells, (ii) during long term immune response to the epitopes, primarily the IgG1 isotype was produced, (iii) in vitro, macrophage degradation of r-GV containing different SIVsm inserts occurred over a period of days resulting in an inherent slow breakdown and degradation of the SIVsm peptide inserts, (iv) vesicle specific GvpC, a larger protein, degraded more slowly than the recombinant peptide inserts and (v) in vitro uptake and degradation of the r-GV populations tested was associated with SIVsm insert specific patterns for cytokines IL-10, IL-12 and IL-18. Conclusions Together these findings provide new information underscoring r-GV potential. They can clearly

  1. Optimizing High Level Waste Disposal

    SciTech Connect

    Dirk Gombert

    2005-09-01

    If society is ever to reap the potential benefits of nuclear energy, technologists must close the fuel-cycle completely. A closed cycle equates to a continued supply of fuel and safe reactors, but also reliable and comprehensive closure of waste issues. High level waste (HLW) disposal in borosilicate glass (BSG) is based on 1970s era evaluations. This host matrix is very adaptable to sequestering a wide variety of radionuclides found in raffinates from spent fuel reprocessing. However, it is now known that the current system is far from optimal for disposal of the diverse HLW streams, and proven alternatives are available to reduce costs by billions of dollars. The basis for HLW disposal should be reassessed to consider extensive waste form and process technology research and development efforts, which have been conducted by the United States Department of Energy (USDOE), international agencies and the private sector. Matching the waste form to the waste chemistry and using currently available technology could increase the waste content in waste forms to 50% or more and double processing rates. Optimization of the HLW disposal system would accelerate HLW disposition and increase repository capacity. This does not necessarily require developing new waste forms, the emphasis should be on qualifying existing matrices to demonstrate protection equal to or better than the baseline glass performance. Also, this proposed effort does not necessarily require developing new technology concepts. The emphasis is on demonstrating existing technology that is clearly better (reliability, productivity, cost) than current technology, and justifying its use in future facilities or retrofitted facilities. Higher waste processing and disposal efficiency can be realized by performing the engineering analyses and trade-studies necessary to select the most efficient methods for processing the full spectrum of wastes across the nuclear complex. This paper will describe technologies being

  2. Guanidine alkaloid analogs as inhibitors of HIV-1 Nef interactions with p53, actin, and p56lck.

    PubMed

    Olszewski, Allison; Sato, Ken; Aron, Zachary D; Cohen, Frederick; Harris, Aleishia; McDougall, Brenda R; Robinson, W Edward; Overman, Larry E; Weiss, Gregory A

    2004-09-28

    With current anti-HIV treatments targeting only 4 of the 15 HIV proteins, many potential viral vulnerabilities remain unexploited. We report small-molecule inhibitors of the HIV-1 protein Nef. In addition to expanding the anti-HIV arsenal, small-molecule inhibitors against untargeted HIV proteins could be used to dissect key events in the HIV lifecycle. Numerous incompletely characterized interactions between Nef and cellular ligands, for example, present a challenge to understanding molecular events during HIV progression to AIDS. Assays with phage-displayed Nef from HIV(NL4-3) were used to identify a series of guanidine alkaloid-based inhibitors of Nef interactions with p53, actin, and p56(lck). The guanidines, synthetic analogs of batzellidine and crambescidin natural products, inhibit the Nef-ligand interactions with IC(50) values in the low micromolar range. In addition, sensitive in vivo assays for Nef inhibition are reported. Although compounds that are effective in vitro proved to be too cytotoxic for cellular assays, the reported Nef inhibitors provide proof-of-concept for disrupting a new HIV target and offer useful leads for drug development.

  3. Differential signaling mechanism for HIV-1 Nef-mediated production of IL-6 and IL-8 in human astrocytes.

    PubMed

    Liu, Xun; Kumar, Anil

    2015-06-15

    Variety of HIV-1 viral proteins including HIV-1 Nef are known to activate astrocytes and microglia in the brain and cause the release of pro-inflammatory cytokines, which is thought to be one of the mechanisms leading to HIV-1- mediated neurotoxicity. IL-6 and IL-8 have been found in the CSF of patients with HIV-1 associated dementia (HAD), suggesting that they might play important roles in HIV-1 neuropathology. In the present study we examined the effects of HIV-1 Nef on IL-6 and IL-8 induction in astrocytes. The results demonstrate that both IL-6 and IL-8 are significantly induced in HIV-1 Nef-transfected SVGA astrocytes and HIV-1 Nef-treated primary fetal astrocytes. We also determined the molecular mechanisms responsible for the HIV-1 Nef-induced increased IL-6 and IL-8 by using chemical inhibitors and siRNAs against PI3K/Akt/PKC, p38 MAPK, NF-κB, CEBP and AP-1. Our results clearly demonstrate that the PI3K/PKC, p38 MAPK, NF-κB and AP-1 pathways are involved in HIV-1 Nef-induced IL-6 production in astrocytes, while PI3K/PKC and NF-κB pathways are involved in HIV-1 Nef-induced IL-8 production. These results offer new potential targets to develop therapeutic strategy for treatment of HIV-1 associated neurological disorders, prevalent in > 40% of individuals infected with HIV-1.

  4. HIV-1 Nef Is Transferred from Expressing T Cells to Hepatocytic Cells through Conduits and Enhances HCV Replication

    PubMed Central

    Park, In-Woo; Fan, Yan; Luo, Xiaoyu; Ryou, Myoung-Gwi; Liu, Jinfeng; Green, Linden; He, Johnny J.

    2014-01-01

    HIV-1 infection enhances HCV replication and as a consequence accelerates HCV-mediated hepatocellular carcinoma (HCC). However, the precise molecular mechanism by which this takes place is currently unknown. Our data showed that infectious HIV-1 failed to replicate in human hepatocytic cell lines. No discernible virus replication was observed, even when the cell lines transfected with HIV-1 proviral DNA were co-cultured with Jurkat T cells, indicating that the problem of liver deterioration in the co-infected patient is not due to the replication of HIV-1 in the hepatocytes of the HCV infected host. Instead, HIV-1 Nef protein was transferred from nef-expressing T cells to hepatocytic cells through conduits, wherein up to 16% (average 10%) of the cells harbored the transferred Nef, when the hepatocytic cells were co-cultured with nef-expressing Jurkat cells for 24 h. Further, Nef altered the size and numbers of lipid droplets (LD), and consistently up-regulated HCV replication by 1.5∼2.5 fold in the target subgenomic replicon cells, which is remarkable in relation to the initially indolent viral replication. Nef also dramatically augmented reactive oxygen species (ROS) production and enhanced ethanol-mediated up-regulation of HCV replication so as to accelerate HCC. Taken together, these data indicate that HIV-1 Nef is a critical element in accelerating progression of liver pathogenesis via enhancing HCV replication and coordinating modulation of key intra- and extra-cellular molecules for liver decay. PMID:24911518

  5. Cell-based Fluorescence Complementation Reveals a Role for HIV-1 Nef Protein Dimerization in AP-2 Adaptor Recruitment and CD4 Co-receptor Down-regulation.

    PubMed

    Shu, Sherry T; Emert-Sedlak, Lori A; Smithgall, Thomas E

    2017-02-17

    The HIV-1 Nef accessory factor enhances viral infectivity, immune evasion, and AIDS progression. Nef triggers rapid down-regulation of CD4 via the endocytic adaptor protein 2 (AP-2) complex, a process linked to enhanced viral infectivity and immune escape. Here, we describe a bimolecular fluorescence complementation (BiFC) assay to visualize the interaction of Nef with AP-2 and CD4 in living cells. Interacting protein pairs were fused to complementary non-fluorescent fragments of YFP and co-expressed in 293T cells. Nef interactions with both CD4 and AP-2 resulted in complementation of YFP and a bright fluorescent signal by confocal microcopy that localized to the cell periphery. Co-expression of the AP-2 α subunit enhanced the Nef·AP-2 σ2 subunit BiFC signal and vice versa, suggesting that the AP-2 α-σ2 hemicomplex interacts cooperatively with Nef. Mutagenesis of Nef amino acids Arg-134, Glu-174, and Asp-175, which stabilize Nef for AP-2 α-σ2 binding in a recent co-crystal structure, substantially reduced AP-2 interaction without affecting CD4 binding. A dimerization-defective mutant of Nef failed to interact with either CD4 or AP-2 in the BiFC assay, indicating that Nef quaternary structure is required for CD4 and AP-2 recruitment as well as CD4 down-regulation. A small molecule previously shown to bind the Nef dimerization interface also reduced Nef interactions with AP-2 and CD4 and restored CD4 expression to the surface of HIV-infected cells. Our findings provide a mechanistic explanation for previous observations that dimerization-defective Nef mutants fail to down-regulate CD4 and validate the Nef dimerization interface as a target site for antiretroviral drug development.

  6. Nef from primary isolates of human immunodeficiency virus type 1 suppresses surface CD4 expression in human and mouse T cells.

    PubMed Central

    Anderson, S; Shugars, D C; Swanstrom, R; Garcia, J V

    1993-01-01

    The human immunodeficiency virus type 1 (HIV-1) nef gene was originally described as a negative regulator of transcription from the viral long terminal repeat promoter. This observation has been disputed, and the function of Nef remains unclear. In vivo experiments have indicated that an intact nef gene is required for disease progression in macaques infected with simian immunodeficiency virus, suggesting a role for Nef in the pathogenesis of AIDS. We and others have previously shown that expression of Nef in cells bearing surface CD4 results in a sustained decrease in surface CD4 expression. This was demonstrated for Nef from two laboratory strains of HIV-1, Bru and SF2. Because both of these isolates were passaged in vitro prior to molecular cloning and in vitro passage can result in mutations which might alter nef gene function, we have analyzed two primary isolates of Nef for their ability to suppress cell surface CD4 expression. The nef genes of HIV-1 isolates from two patients with fewer than 200 CD4+ T cells per mm3 of blood were introduced into human and mouse T-cell lines by retrovirus-mediated gene transfer. Expression of Nef from both isolates correlated with a decrease in surface expression of both human and mouse CD4. To determine whether the ability to suppress surface CD4 expression is a general function of Nef, we also tested an artificially generated consensus nef gene derived from analysis of 54 patient isolates of HIV-1. Expression of the consensus Nef protein also correlated with decreased cell surface CD4 expression in both mouse and human T-cell lines. These results suggest that the ability to suppress cell surface CD4 expression is an intrinsic feature of HIV-1 Nef. Images PMID:8331733

  7. HIV-1 Nef hijacks clathrin coats by stabilizing AP-1:Arf1 polygons.

    PubMed

    Shen, Qing-Tao; Ren, Xuefeng; Zhang, Rui; Lee, Il-Hyung; Hurley, James H

    2015-10-23

    The lentiviruses HIV and simian immunodeficiency virus (SIV) subvert intracellular membrane traffic as part of their replication cycle. The lentiviral Nef protein helps viruses evade innate and adaptive immune defenses by hijacking the adaptor protein 1 (AP-1) and AP-2 clathrin adaptors. We found that HIV-1 Nef and the guanosine triphosphatase Arf1 induced trimerization and activation of AP-1. Here we report the cryo-electron microscopy structures of the Nef- and Arf1-bound AP-1 trimer in the active and inactive states. A central nucleus of three Arf1 molecules organizes the trimers. We combined the open trimer with a known dimer structure and thus predicted a hexagonal assembly with inner and outer faces that bind the membranes and clathrin, respectively. Hexagons were directly visualized and the model validated by reconstituting clathrin cage assembly. Arf1 and Nef thus play interconnected roles in allosteric activation, cargo recruitment, and coat assembly, revealing an unexpectedly intricate organization of the inner AP-1 layer of the clathrin coat.

  8. High temperature liquid level sensor

    DOEpatents

    Tokarz, Richard D.

    1983-01-01

    A length of metal sheathed metal oxide cable is perforated to permit liquid access to the insulation about a pair of conductors spaced close to one another. Changes in resistance across the conductors will be a function of liquid level, since the wetted insulation will have greater electrical conductivity than that of the dry insulation above the liquid elevation.

  9. The HIV proteins Tat and Nef promote human bone marrow mesenchymal stem cell senescence and alter osteoblastic differentiation.

    PubMed

    Beaupere, Carine; Garcia, Marie; Larghero, Jerome; Fève, Bruno; Capeau, Jacqueline; Lagathu, Claire

    2015-08-01

    To maintain bone mass turnover and bone mineral density (BMD), bone marrow (BM) mesenchymal stem cells (MSCs) are constantly recruited and subsequently differentiated into osteoblasts. HIV-infected patients present lower BMD than non-HIV infected individuals and a higher prevalence of osteopenia/osteoporosis. In antiretroviral treatment (ART)-naive patients, encoded HIV proteins represent pathogenic candidates. They are released by infected cells within BM and can impact on neighbouring cells. In this study, we tested whether HIV proteins Tat and/or Nef could induce senescence of human BM-MSCs and reduce their capacity to differentiate into osteoblasts. When compared to nontreated cells, MSCs chronically treated with Tat and/or Nef up to 30 days reduced their proliferative activity and underwent early senescence, associated with increased oxidative stress and mitochondrial dysfunction. The antioxidant molecule N-acetyl- cysteine had no or minimal effects on Tat- or Nef-induced senescence. Tat but not Nef induced an early increase in NF-κB activity and cytokine/chemokine secretion. Tat-induced effects were prevented by the NF-κB inhibitor parthenolide, indicating that Tat triggered senescence via NF-κB activation leading to oxidative stress. Otherwise, Nef- but not Tat-treated cells displayed early inhibition of autophagy. Rapamycin, an autophagy inducer, reversed Nef-induced senescence and oxidative stress. Moreover, Tat+Nef had cumulative effects. Finally, Tat and/or Nef decreased the MSC potential of osteoblastic differentiation. In conclusion, our in vitro data show that Tat and Nef could reduce the number of available precursors by inducing MSC senescence, through either enhanced inflammation or reduced autophagy. These results offer new insights into the pathophysiological mechanisms of decreased BMD in HIV-infected patients.

  10. The HIV proteins Tat and Nef promote human bone marrow mesenchymal stem cell senescence and alter osteoblastic differentiation

    PubMed Central

    Beaupere, Carine; Garcia, Marie; Larghero, Jerome; Fève, Bruno; Capeau, Jacqueline; Lagathu, Claire

    2015-01-01

    To maintain bone mass turnover and bone mineral density (BMD), bone marrow (BM) mesenchymal stem cells (MSCs) are constantly recruited and subsequently differentiated into osteoblasts. HIV-infected patients present lower BMD than non-HIV infected individuals and a higher prevalence of osteopenia/osteoporosis. In antiretroviral treatment (ART)-naive patients, encoded HIV proteins represent pathogenic candidates. They are released by infected cells within BM and can impact on neighbouring cells. In this study, we tested whether HIV proteins Tat and/or Nef could induce senescence of human BM-MSCs and reduce their capacity to differentiate into osteoblasts. When compared to nontreated cells, MSCs chronically treated with Tat and/or Nef up to 30 days reduced their proliferative activity and underwent early senescence, associated with increased oxidative stress and mitochondrial dysfunction. The antioxidant molecule N-acetyl- cysteine had no or minimal effects on Tat- or Nef-induced senescence. Tat but not Nef induced an early increase in NF-κB activity and cytokine/chemokine secretion. Tat-induced effects were prevented by the NF-κB inhibitor parthenolide, indicating that Tat triggered senescence via NF-κB activation leading to oxidative stress. Otherwise, Nef- but not Tat-treated cells displayed early inhibition of autophagy. Rapamycin, an autophagy inducer, reversed Nef-induced senescence and oxidative stress. Moreover, Tat+Nef had cumulative effects. Finally, Tat and/or Nef decreased the MSC potential of osteoblastic differentiation. In conclusion, our in vitro data show that Tat and Nef could reduce the number of available precursors by inducing MSC senescence, through either enhanced inflammation or reduced autophagy. These results offer new insights into the pathophysiological mechanisms of decreased BMD in HIV-infected patients. PMID:25847297

  11. High-Level Connectionist Models

    DTIC Science & Technology

    1990-04-01

    Freeman, 1987) and on the mathematical level ( Derrida & Meir, 1988; Huberman & Hogg, 1987; Kurten, 1987). It is time that this link be further...Wesley. Derrida B. & Meir, R. (198sL80moac behavior of a layered neural network. Phys. Rev. A, 38. Elman, J. L. (1988). Findi4 Structure in Time. Report...and Huberman, 1983; Kurten and Clark. 1986; Babcock and Westervelt, 1987; Derrida and Meir, 1988; Riedal et al., 1988; Sompolin- sky et al., 1988

  12. gag, vif, and nef Genes Contribute to the Homologous Viral Interference Induced by a Nonproducer Human Immunodeficiency Virus Type 1 (HIV-1) Variant: Identification of Novel HIV-1-Inhibiting Viral Protein Mutants

    PubMed Central

    D’Aloja, Paola; Olivetta, Eleonora; Bona, Roberta; Nappi, Filomena; Pedacchia, Daniela; Pugliese, Katherina; Ferrari, Giuliana; Verani, Paola; Federico, Maurizio

    1998-01-01

    We previously demonstrated that expression of the nonproducer F12-human immunodeficiency virus type 1 (HIV-1) variant induces a block in the replication of superinfecting HIV that does not depend on the down-regulation of CD4 HIV receptors. In order to individuate the gene(s) involved in F12-HIV-induced interference, vectors expressing each of the nine F12-HIV proteins were transfected in HIV-susceptible HeLa CD4 cells. Pools of cell clones stably producing each viral protein were infected with HIV-1, and virus release was measured in terms of reverse transcriptase activity in supernatants. We hereby demonstrate that HeLa CD4 cells expressing the F12-HIV gag, vif, or nef gene were resistant, to different degrees, to infection with T-cell-line-adapted HIV-1 strains. Conversely, expression of either the tat, rev, or vpu F12-HIV gene increased the rate of HIV release, and no apparent effects on HIV replication were observed in cells expressing either the F12-HIV vpr, pol, or env gene. No variation of CD4 exposure was detected in any of the uninfected HeLa CD4 pools. These data indicate that F12-HIV homologous viral interference is the consequence of the synergistic anti-HIV effects of Gag, Vif, and Nef proteins. Retrovirus vectors expressing F12-HIV vif or nef allowed us to further establish that the expression of each mutated protein (i) inhibits the replication of clinical HIV-1 isolates as well, (ii) impairs the infectivity of the virus released by cells chronically infected with HIV-1, and (iii) limitedly to F12-HIV Vif protein, induces HIV resistance in both vif-permissive and vif-nonpermissive cells. The levels of action of F12-HIV vif and nef anti-HIV effects were also determined. We observed that HIV virions emerging from the first viral cycle on F12-HIV vif-expressing cells, although released in unaltered amounts, had a strongly reduced ability to initiate the retrotranscription process when they reinfected parental HeLa CD4 cells. Differently, we observed

  13. High-Level Data Races

    NASA Technical Reports Server (NTRS)

    Artho, Cyrille; Havelund, Klaus; Biere, Armin; Koga, Dennis (Technical Monitor)

    2003-01-01

    Data races are a common problem in concurrent and multi-threaded programming. They are hard to detect without proper tool support. Despite the successful application of these tools, experience shows that the notion of data race is not powerful enough to capture certain types of inconsistencies occurring in practice. In this paper we investigate data races on a higher abstraction layer. This enables us to detect inconsistent uses of shared variables, even if no classical race condition occurs. For example, a data structure representing a coordinate pair may have to be treated atomically. By lifting the meaning of a data race to a higher level, such problems can now be covered. The paper defines the concepts view and view consistency to give a notation for this novel kind of property. It describes what kinds of errors can be detected with this new definition, and where its limitations are. It also gives a formal guideline for using data structures in a multi-threading environment.

  14. HIV’s Nef Interacts with β-Catenin of the Wnt Signaling Pathway in HEK293 Cells

    PubMed Central

    Weiser, Keren; Barton, Meredith; Gershoony, Dafna; DasGupta, Ramanuj; Cardozo, Timothy

    2013-01-01

    The Wnt signaling pathway is implicated in major physiologic cellular functions, such as proliferation, migration, cell fate specification, maintenance of pluripotency and induction of tumorigenicity. Proliferation and migration are important responses of T-cells, which are major cellular targets of HIV infection. Using an informatics screen, we identified a previously unsuspected interaction between HIV’s Nef protein and β-catenin, a key component of the Wnt pathway. A segment in Nef contains identical amino acids at key positions and structurally mimics the β-catenin binding sites on endogenous β-catenin ligands. The interaction between Nef and β-catenin was confirmed in vitro and in a co-immunoprecipitation from HEK293 cells. Moreover, the introduction of Nef into HEK293 cells specifically inhibited a Wnt pathway reporter. PMID:24130899

  15. Dissociation of the CD4 downregulation and viral infectivity enhancement functions of human immunodeficiency virus type 1 Nef.

    PubMed Central

    Goldsmith, M A; Warmerdam, M T; Atchison, R E; Miller, M D; Greene, W C

    1995-01-01

    Recent evidence indicates that the nef gene of human immunodeficiency virus type 1 augments rather than inhibits viral replication in both cell culture and in vivo models. In addition, nef alters various normal cellular processes, including the display of CD4 on the cell surface. However, it remains unknown whether the enhancement of infectivity and the downregulation of CD4 represent linked or independent biologic properties of this single protein. In the present studies, mutational analyses were performed to define structure-function relationships within the Nef protein that mediate these effects. To assess the functional consequences of these mutations, sensitive and reliable assays were developed to quantitate the viral infectivity enhancement and CD4 downregulation functions of Nef. The results indicate that membrane-targeting sequences at the N terminus of Nef are important for both functions of Nef, while certain other conserved regions are dispensable for both functions. A conserved proline-X-X repeat segment in the central core of the protein, which is reminiscent of an SH3-binding domain, is critical for the enhancement of infectivity function but is dispensable for CD4 downregulation. However, the downregulation of CD4 by Nef appears to involve a two-step process requiring the initial dissociation of p56lck from CD4 to permit engagement of the endocytic apparatus by CD4. Together, these findings demonstrate that the infectivity enhancement and CD4 downregulation activities of human immunodeficiency virus type 1 Nef can be dissociated. Thus, these processes may be independent of one another in the viral replication cycle. PMID:7769669

  16. Brain transcriptome-wide screen for HIV-1 Nef protein interaction partners reveals various membrane-associated proteins.

    PubMed

    Kammula, Ellen C; Mötter, Jessica; Gorgels, Alexandra; Jonas, Esther; Hoffmann, Silke; Willbold, Dieter

    2012-01-01

    HIV-1 Nef protein contributes essentially to the pathology of AIDS by a variety of protein-protein-interactions within the host cell. The versatile functionality of Nef is partially attributed to different conformational states and posttranslational modifications, such as myristoylation. Up to now, many interaction partners of Nef have been identified using classical yeast two-hybrid screens. Such screens rely on transcriptional activation of reporter genes in the nucleus to detect interactions. Thus, the identification of Nef interaction partners that are integral membrane proteins, membrane-associated proteins or other proteins that do not translocate into the nucleus is hampered. In the present study, a split-ubiquitin based yeast two-hybrid screen was used to identify novel membrane-localized interaction partners of Nef. More than 80% of the hereby identified interaction partners of Nef are transmembrane proteins. The identified hits are GPM6B, GPM6A, BAP31, TSPAN7, CYB5B, CD320/TCblR, VSIG4, PMEPA1, OCIAD1, ITGB1, CHN1, PH4, CLDN10, HSPA9, APR-3, PEBP1 and B3GNT, which are involved in diverse cellular processes like signaling, apoptosis, neurogenesis, cell adhesion and protein trafficking or quality control. For a subfraction of the hereby identified proteins we present data supporting their direct interaction with HIV-1 Nef. We discuss the results with respect to many phenotypes observed in HIV infected cells and patients. The identified Nef interaction partners may help to further elucidate the molecular basis of HIV-related diseases.

  17. Nef-M1, a peptide antagonist of CXCR4, inhibits tumor angiogenesis and epithelial‑to‑mesenchymal transition in colon and breast cancers.

    PubMed

    Katkoori, Venkat R; Basson, Marc D; Bond, Vincent C; Manne, Upender; Bumpers, Harvey L

    2015-09-29

    The Nef-M1 peptide competes effectively with the natural ligand of CXC chemokine receptor 4 (CXCR4), stromal cell-derived factor 1-alpha, to induce apoptosis and inhibit growth in colon cancer (CRC) and breast cancer (BC). Its role in tumor angiogenesis, and epithelial-to-mesenchymal transition (EMT) regulation, key steps involved in tumor growth and metastasis, are unknown. We evaluated the angioinhibitory effect of Nef-M1 peptide and examined its role in the inhibition of EMT in these cancers. Colon (HT29) and breast (MDA-MB231) cancer cells expressing CXCR4 were studied in vitro and in xenograft tumors propagated in severe combined immunodeficient mice. The mice were treated intraperitoneally with Nef-M1 or scrambled amino acid sequence of Nef-M1 (sNef-M1) peptide, a negative control, starting at the time of tumor implantation. Sections from tumors were evaluated for tumor angiogenesis, as measured by microvessel density (MVD) based on immunostaining of endothelial markers. In vitro tumor angiogenesis was assessed by treating human umbilical vein endothelial cells with conditioned media from the tumor cell lines. A BC cell line (MDA-MB 468) which does not express CXCR4 was used to study the actions of Nef-M1 peptide. Western blot and immunofluorescence analyses assessed the effect of Nef-M1 on tumor angiogenesis and EMT in both tumors and cancer cells. Metastatic lesions of CRC and BC expressed more CXCR4 than primary lesions. It was also found that tumors from mice treated with sNef-M1 had well established vascularity, while Nef-M1 treated tumors had very poor vascularization. Indeed, the mean MVD was lower in tumors from Nef-M1 treated mice than in sNef-M1 treated tumors. Nef-M1 treated tumor has poor morphology and loss of endothelial integrity. Although conditioned medium from CRC or BC cells supported HUVEC tube formation, the conditioned medium from Nef-M1 treated CRC or BC cells did not support tube formation. Western blot analyses revealed that Nef-M1

  18. Guanidine alkaloid analogs as inhibitors of HIV-1 Nef interactions with p53, actin, and p56lck

    PubMed Central

    Olszewski, Allison; Sato, Ken; Aron, Zachary D.; Cohen, Frederick; Harris, Aleishia; McDougall, Brenda R.; Robinson, W. Edward; Overman, Larry E.; Weiss, Gregory A.

    2004-01-01

    With current anti-HIV treatments targeting only 4 of the 15 HIV proteins, many potential viral vulnerabilities remain unexploited. We report small-molecule inhibitors of the HIV-1 protein Nef. In addition to expanding the anti-HIV arsenal, small-molecule inhibitors against untargeted HIV proteins could be used to dissect key events in the HIV lifecycle. Numerous incompletely characterized interactions between Nef and cellular ligands, for example, present a challenge to understanding molecular events during HIV progression to AIDS. Assays with phage-displayed Nef from HIVNL4-3 were used to identify a series of guanidine alkaloid-based inhibitors of Nef interactions with p53, actin, and p56lck. The guanidines, synthetic analogs of batzellidine and crambescidin natural products, inhibit the Nef–ligand interactions with IC50 values in the low micromolar range. In addition, sensitive in vivo assays for Nef inhibition are reported. Although compounds that are effective in vitro proved to be too cytotoxic for cellular assays, the reported Nef inhibitors provide proof-of-concept for disrupting a new HIV target and offer useful leads for drug development. PMID:15371598

  19. Higher cross-subtype IFN-γ ELISpot responses to Gag and Nef peptides in Brazilian HIV-1 subtype B- and F1- than in C-infected subjects.

    PubMed

    Côrtes, Fernanda Heloise; Bello, Gonzalo; Vorsatz, Carla; Pilotto, José Henrique; Guimarães, Monick Lindenmeyer; Grinsztejn, Beatriz; Veloso, Valdiléa Gonçalves; Pinto, Aguinaldo Roberto; Morgado, Mariza Gonçalves

    2013-02-04

    HIV-1 diversity has been considered a huge challenge for the HIV-1 vaccine development. To overcome it, immunogens based on centralized sequences, as consensus, have been tested. In Brazil, the co-circulation of three subtypes offers a suitable scenario to test T cell cross-subtype responses to consensus sequences. Furthermore, we included peptides based on closest viral isolates (CVI) from each subtype analyzed to compare with T cell responses detected against the consensus sequences. The study included 32 subjects infected with HIV-1 subtype B (n=13),C (n=11), and F1 (n=8). Gag and Nef-specific T cell responses were evaluated by IFN-γ-ELISpot assay. Peptides based on CVI sequences were similar to consensus in both reducing genetic distance and detecting T cell responses. A high cross-subtype response between B and F1 in both regions was observed in HIV-1 subtype B and F1-infected subjects. We also found no significant difference in responses to subtype B and C consensus peptides among subtype B-infected subjects. In contrast, the magnitude of T cell responses to consensus C peptides in the Gag region was higher than to consensus B peptides among HIV-1 subtype C-infected subjects. Regarding Nef, subtype C-infected subjects showed higher values to consensus C than to consensus F1 peptides. Moreover, subtype F1-infected subjects presented lower responses to subtype C peptides than to subtype F1 and B. A similar level of responses was detected with group M based peptides in subtype B and F1 infected subjects. However, among subtype C infected subjects, this set of peptides detected lower levels of response than consensus C. Overall, the level of cross-subtype response between subtypes B and F1 was higher than between subtype C and B or C and F1. Our data suggests that the barrier of genetic diversity in HIV-1 group M for vaccine design may be dependent on the subtypes involved. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Association of the dopamine receptor interacting protein gene, NEF3, with early response to antipsychotic medication.

    PubMed

    Strous, Rael D; Greenbaum, Lior; Kanyas, Kyra; Merbl, Yifat; Horowitz, Anat; Karni, Osnat; Viglin, Dina; Olender, Tsviya; Deshpande, Smita N; Lancet, Doron; Ben-Asher, Edna; Lerer, Bernard

    2007-06-01

    Genetic variation in antipsychotic drug targets could underlie variability among patients in the time required for antipsychotic effects to be elicited. In a clinical, pharmacogenetic study we focused on the dopamine receptor interacting protein (DRIP) gene family. DRIPs are pivotally involved in regulating dopamine receptor signal transduction. Consecutively hospitalized, acutely psychotic patients with DSM-IV schizophrenia (n=121) were included in the study if they received treatment with typical antipsychotic medication (TYP, n=72) or TYP plus risperidone (TYP-R, n=49) for at least 2 wk. Clinical state and adverse effects were rated at baseline and after 2 wk. Patients improved significantly on both TYP and TYP-R with no significant difference between them. Early responders were defined as patients whose PANSS change scores were greater than the median. Twenty-two single nucleotide polymorphisms (SNPs) were analysed in five DRIP-encoding genes. Two SNPs in NEF3, which encodes the DRIP, neurofilament-medium (NF-M), were associated with early response (rs1457266, p=0.01; rs1379357, p=0.006). A 5 SNP haplotype spanning NEF3 was over-represented in early responders (p=0.015), in the combined patient group and in the TYP group alone. These findings suggest that variation in NEF3, most likely functional variants that are in linkage disequilibrium with the SNPs that we studied, influences rate of response to TYP. Since NEF3 is primarily associated with dopamine D1 receptor function, the evidence for a complementary role of dopamine D1 receptors in antipsychotic effects is considered. The findings reported here open an interesting research avenue in the pharmacogenetics of antipsychotic effects but require replication in larger samples treated in a controlled context.

  1. Activation/proliferation and apoptosis of bystander goat lymphocytes induced by a macrophage-tropic chimeric caprine arthritis encephalitis virus expressing SIV Nef

    SciTech Connect

    Bouzar, Baya Amel; Rea, Angela; Hoc-Villet, Stephanie; Garnier, Celine; Guiguen, Francois; Jin Yuhuai; Narayan, Opendra; Chebloune, Yahia . E-mail: ychebloune@kumc.edu

    2007-08-01

    Caprine arthritis encephalitis virus (CAEV) is the natural lentivirus of goats, well known for its tropism for macrophages and its inability to cause infection in lymphocytes. The viral genome lacks nef, tat, vpu and vpx coding sequences. To test the hypothesis that when nef is expressed by the viral genome, the virus became toxic for lymphocytes during replication in macrophages, we inserted the SIVsmm PBj14 nef coding sequences into the genome of CAEV thereby generating CAEV-nef. This recombinant virus is not infectious for lymphocytes but is fully replication competent in goat macrophages in which it constitutively expresses the SIV Nef. We found that goat lymphocytes cocultured with CAEV-nef-infected macrophages became activated, showing increased expression of the interleukin-2 receptor (IL-2R). Activation correlated with increased proliferation of the cells. Interestingly, a dual effect in terms of apoptosis regulation was observed in exposed goat lymphocytes. Nef was found first to induce a protection of lymphocytes from apoptosis during the first few days following exposure to infected macrophages, but later it induced increased apoptosis in the activated lymphocytes. This new recombinant virus provides a model to study the functions of Nef in the context of infection of macrophages, but in absence of infection of T lymphocytes and brings new insights into the biological effects of Nef on lymphocytes.

  2. HIV-1 Nef assembles a Src family kinase-ZAP-70/Syk-PI3K cascade to downregulate cell-surface MHC-I.

    PubMed

    Hung, Chien-Hui; Thomas, Laurel; Ruby, Carl E; Atkins, Katelyn M; Morris, Nicholas P; Knight, Zachary A; Scholz, Isabel; Barklis, Eric; Weinberg, Andrew D; Shokat, Kevan M; Thomas, Gary

    2007-04-19

    HIV-1 Nef, which is required for the efficient onset of AIDS, enhances viral replication and infectivity by exerting multiple effects on infected cells. Nef downregulates cell-surface MHC-I molecules by an uncharacterized PI3K pathway requiring the actions of two Nef motifs-EEEE(65) and PXXP(75). We report that the Nef EEEE(65) targeting motif enables Nef PXXP(75) to bind and activate a trans-Golgi network-localized Src family tyrosine kinase (SFK). The Nef/SFK complex then recruits and phosphorylates the tyrosine kinase ZAP-70, which binds class I PI3K to trigger MHC-I downregulation in primary CD4+ T cells. In promonocytic cells, Nef/SFK recruits the ZAP-70 homolog Syk to downregulate MHC-I, implicating this PI3K pathway in multiple HIV-1 reservoirs. Isoform-specific PI3K inhibitors repress MHC-I downregulation, identifying them as potential therapeutic agents to combat HIV-1. The discovery of this Nef-SFK-ZAP-70/Syk-PI3K signaling pathway explains the hierarchal role of the Nef motifs in effecting immunoevasion.

  3. Remodeling of the Host Cell Plasma Membrane by HIV-1 Nef and Vpu: A Strategy to Ensure Viral Fitness and Persistence

    PubMed Central

    Sugden, Scott M.; Bego, Mariana G.; Pham, Tram N.Q.; Cohen, Éric A.

    2016-01-01

    The plasma membrane protects the cell from its surroundings and regulates cellular communication, homing, and metabolism. Not surprisingly, the composition of this membrane is highly controlled through the vesicular trafficking of proteins to and from the cell surface. As intracellular pathogens, most viruses exploit the host plasma membrane to promote viral replication while avoiding immune detection. This is particularly true for the enveloped human immunodeficiency virus (HIV), which assembles and obtains its lipid shell directly at the plasma membrane. HIV-1 encodes two proteins, negative factor (Nef) and viral protein U (Vpu), which function primarily by altering the quantity and localization of cell surface molecules to increase virus fitness despite host antiviral immune responses. These proteins are expressed at different stages in the HIV-1 life cycle and employ a variety of mechanisms to target both unique and redundant surface proteins, including the viral receptor CD4, host restriction factors, immunoreceptors, homing molecules, tetraspanins and membrane transporters. In this review, we discuss recent progress in the study of the Nef and Vpu targeting of host membrane proteins with an emphasis on how remodeling of the cell membrane allows HIV-1 to avoid host antiviral immune responses leading to the establishment of systemic and persistent infection. PMID:26950141

  4. Remodeling of the Host Cell Plasma Membrane by HIV-1 Nef and Vpu: A Strategy to Ensure Viral Fitness and Persistence.

    PubMed

    Sugden, Scott M; Bego, Mariana G; Pham, Tram N Q; Cohen, Éric A

    2016-03-03

    The plasma membrane protects the cell from its surroundings and regulates cellular communication, homing, and metabolism. Not surprisingly, the composition of this membrane is highly controlled through the vesicular trafficking of proteins to and from the cell surface. As intracellular pathogens, most viruses exploit the host plasma membrane to promote viral replication while avoiding immune detection. This is particularly true for the enveloped human immunodeficiency virus (HIV), which assembles and obtains its lipid shell directly at the plasma membrane. HIV-1 encodes two proteins, negative factor (Nef) and viral protein U (Vpu), which function primarily by altering the quantity and localization of cell surface molecules to increase virus fitness despite host antiviral immune responses. These proteins are expressed at different stages in the HIV-1 life cycle and employ a variety of mechanisms to target both unique and redundant surface proteins, including the viral receptor CD4, host restriction factors, immunoreceptors, homing molecules, tetraspanins and membrane transporters. In this review, we discuss recent progress in the study of the Nef and Vpu targeting of host membrane proteins with an emphasis on how remodeling of the cell membrane allows HIV-1 to avoid host antiviral immune responses leading to the establishment of systemic and persistent infection.

  5. High-Level Binocular Rivalry Effects

    PubMed Central

    Wolf, Michal; Hochstein, Shaul

    2011-01-01

    Binocular rivalry (BR) occurs when the brain cannot fuse percepts from the two eyes because they are different. We review results relating to an ongoing controversy regarding the cortical site of the BR mechanism. Some BR qualities suggest it is low-level: (1) BR, as its name implies, is usually between eyes and only low-levels have access to utrocular information. (2) All input to one eye is suppressed: blurring doesn’t stimulate accommodation; pupilary constrictions are reduced; probe detection is reduced. (3) Rivalry is affected by low-level attributes, contrast, spatial frequency, brightness, motion. (4) There is limited priming due to suppressed words or pictures. On the other hand, recent studies favor a high-level mechanism: (1) Rivalry occurs between patterns, not eyes, as in patchwork rivalry or a swapping paradigm. (2) Attention affects alternations. (3) Context affects dominance. There is conflicting evidence from physiological studies (single cell and fMRI) regarding cortical level(s) of conscious perception. We discuss the possibility of multiple BR sites and theoretical considerations that rule out this solution. We present new data regarding the locus of the BR switch by manipulating stimulus semantic content or high-level characteristics. Since these variations are represented at higher cortical levels, their affecting rivalry supports high-level BR intervention. In Experiment I, we measure rivalry when one eye views words and the other non-words and find significantly longer dominance durations for non-words. In Experiment II, we find longer dominance times for line drawings of simple, structurally impossible figures than for similar, possible objects. In Experiment III, we test the influence of idiomatic context on rivalry between words. Results show that generally words within their idiomatic context have longer mean dominance durations. We conclude that BR has high-level cortical influences, and may be controlled by a high-level mechanism

  6. High-level binocular rivalry effects.

    PubMed

    Wolf, Michal; Hochstein, Shaul

    2011-01-01

    Binocular rivalry (BR) occurs when the brain cannot fuse percepts from the two eyes because they are different. We review results relating to an ongoing controversy regarding the cortical site of the BR mechanism. Some BR qualities suggest it is low-level: (1) BR, as its name implies, is usually between eyes and only low-levels have access to utrocular information. (2) All input to one eye is suppressed: blurring doesn't stimulate accommodation; pupilary constrictions are reduced; probe detection is reduced. (3) Rivalry is affected by low-level attributes, contrast, spatial frequency, brightness, motion. (4) There is limited priming due to suppressed words or pictures. On the other hand, recent studies favor a high-level mechanism: (1) Rivalry occurs between patterns, not eyes, as in patchwork rivalry or a swapping paradigm. (2) Attention affects alternations. (3) Context affects dominance. There is conflicting evidence from physiological studies (single cell and fMRI) regarding cortical level(s) of conscious perception. We discuss the possibility of multiple BR sites and theoretical considerations that rule out this solution. We present new data regarding the locus of the BR switch by manipulating stimulus semantic content or high-level characteristics. Since these variations are represented at higher cortical levels, their affecting rivalry supports high-level BR intervention. In Experiment I, we measure rivalry when one eye views words and the other non-words and find significantly longer dominance durations for non-words. In Experiment II, we find longer dominance times for line drawings of simple, structurally impossible figures than for similar, possible objects. In Experiment III, we test the influence of idiomatic context on rivalry between words. Results show that generally words within their idiomatic context have longer mean dominance durations. We conclude that BR has high-level cortical influences, and may be controlled by a high-level mechanism.

  7. Characterization of the mammalian RNA exonuclease 5/NEF-sp as a testis-specific nuclear 3' → 5' exoribonuclease.

    PubMed

    Silva, Sara; Homolka, David; Pillai, Ramesh S

    2017-09-01

    Ribonucleases catalyze maturation of functional RNAs or mediate degradation of cellular transcripts, activities that are critical for gene expression control. Here we identify a previously uncharacterized mammalian nuclease family member NEF-sp (RNA exonuclease 5 [REXO5] or LOC81691) as a testis-specific factor. Recombinant human NEF-sp demonstrates a divalent metal ion-dependent 3' → 5' exoribonuclease activity. This activity is specific to single-stranded RNA substrates and is independent of their length. The presence of a 2'-O-methyl modification at the 3' end of the RNA substrate is inhibitory. Ectopically expressed NEF-sp localizes to the nucleolar/nuclear compartment in mammalian cell cultures and this is dependent on an amino-terminal nuclear localization signal. Finally, mice lacking NEF-sp are viable and display normal fertility, likely indicating overlapping functions with other nucleases. Taken together, our study provides the first biochemical and genetic exploration of the role of the NEF-sp exoribonuclease in the mammalian genome. © 2017 Silva et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  8. Mutations in the nef and vif genes associated with progression to AIDS in elite controller and slow-progressor patients.

    PubMed

    Cruz, Nadia V G; Amorim, Raquel; Oliveira, Fátima E; Speranza, Francisco A C; Costa, Luciana J

    2013-04-01

    Progression towards AIDS can vary from 5 to 10 years from the establishment of the primary infection by HIV-1 to more than 10 years in the complete absence of antiretroviral therapy. Several factors can contribute to the outcome of HIV infection, including host genetic and viral replicating characteristics. Historically, nef-deleted viral genomes have been associated with disease progression. Therefore, the lentiviral Nef protein is regarded as a progression factor. The objective of this work was to characterize the nef gene from a group of treatment naive patients infected with HIV-1 for more than 10 years. These patients were classified as long-term non-progressors, elite controller, and slow-progressors according to clinical and laboratorial data. A premature stop codon within the nef gene leading to the expression of a truncated peptide was observed on samples from the elite controller patient. For the slow-progressor patients, several degrees of deletions at the C-terminal of Nef were observed predicting a loss of function of this protein. The vif gene was characterized for these patients and a rare mutation that predicts a miss localization of the Vif protein to the nucleus of infected cells that could prevent its function as an APOBEC neutralization factor was also observed. These data indicate the importance of the HIV accessory proteins as factors that contribute to the outcome of AIDS.

  9. Parallel Processing at the High School Level.

    ERIC Educational Resources Information Center

    Sheary, Kathryn Anne

    This study investigated the ability of high school students to cognitively understand and implement parallel processing. Data indicates that most parallel processing is being taught at the university level. Instructional modules on C, Linux, and the parallel processing language, P4, were designed to show that high school students are highly…

  10. High-Level Application Framework for LCLS

    SciTech Connect

    Chu, P; Chevtsov, S.; Fairley, D.; Larrieu, C.; Rock, J.; Rogind, D.; White, G.; Zalazny, M.; /SLAC

    2008-04-22

    A framework for high level accelerator application software is being developed for the Linac Coherent Light Source (LCLS). The framework is based on plug-in technology developed by an open source project, Eclipse. Many existing functionalities provided by Eclipse are available to high-level applications written within this framework. The framework also contains static data storage configuration and dynamic data connectivity. Because the framework is Eclipse-based, it is highly compatible with any other Eclipse plug-ins. The entire infrastructure of the software framework will be presented. Planned applications and plug-ins based on the framework are also presented.

  11. Cell Surface Proteomic Map of HIV Infection Reveals Antagonism of Amino Acid Metabolism by Vpu and Nef

    PubMed Central

    Matheson, Nicholas J.; Sumner, Jonathan; Wals, Kim; Rapiteanu, Radu; Weekes, Michael P.; Vigan, Raphael; Weinelt, Julia; Schindler, Michael; Antrobus, Robin; Costa, Ana S.H.; Frezza, Christian; Clish, Clary B.; Neil, Stuart J.D.; Lehner, Paul J.

    2015-01-01

    Summary Critical cell surface immunoreceptors downregulated during HIV infection have previously been identified using non-systematic, candidate approaches. To gain a comprehensive, unbiased overview of how HIV infection remodels the T cell surface, we took a distinct, systems-level, quantitative proteomic approach. >100 plasma membrane proteins, many without characterized immune functions, were downregulated during HIV infection. Host factors targeted by the viral accessory proteins Vpu or Nef included the amino acid transporter SNAT1 and the serine carriers SERINC3/5. We focused on SNAT1, a β-TrCP-dependent Vpu substrate. SNAT1 antagonism was acquired by Vpu variants from the lineage of SIVcpz/HIV-1 viruses responsible for pandemic AIDS. We found marked SNAT1 induction in activated primary human CD4+ T cells, and used Consumption and Release (CoRe) metabolomics to identify alanine as an endogenous SNAT1 substrate required for T cell mitogenesis. Downregulation of SNAT1 therefore defines a unique paradigm of HIV interference with immunometabolism. PMID:26439863

  12. Rituximab fails where eculizumab restores renal function in C3nef-related DDD.

    PubMed

    Rousset-Rouvière, Caroline; Cailliez, Mathilde; Garaix, Florentine; Bruno, Daniele; Laurent, Daniel; Tsimaratos, Michel

    2014-06-01

    Dense deposit disease (DDD), a C3 glomerulopathy (C3G), is a rare disease with unfavorable progression towards end-stage kidney disease. The pathogenesis of DDD is due to cytotoxic effects related to acquired or genetic dysregulation of the complement alternative pathway, which is at times accompanied by the production of C3 nephritic factor (C3NeF), an auto-antibody directed against the alternative C3 convertase. Available treatments include plasma exchange, CD20-targeted antibodies, and a terminal complement blockade via the anti-C5 monoclonal antibody eculizumab. We report here the case of an 8-year-old child with C3NeF and refractory DDD who presented with a nephritic syndrome. She tested positive for C3NeF activity; C3 was undetectable. Genetic analyses of the alternative complement pathway were normal. Methylprednisolone pulses and mycophenolate mofetil treatment resulted in complete recovery of renal function and a reduction in proteinuria. Corticosteroids were tapered and then withdrawn. Four months after corticosteroid discontinuation, hematuria and proteinuria recurred, and a renal biopsy confirmed an active DDD with a majority of extracapillary crescents. Despite an increase in immunosuppressive drugs, including methylprednisolone pulses and rituximab therapy, the patient suffered acute renal failure within 3 weeks, requiring dialysis. Eculizumab treatment resulted in a quick and impressive response. Hematuria very quickly resolved, kidney function improved, and no further dialysis was required. The patient received bimonthly eculizumab injections of 600 mg, allowing for normalization of renal function and reduction of proteinuria to <0.5 g per day. Since then, she continues to receive eculizumab. Complement regulation pathway-targeted therapy may be a specific and useful treatment for rapidly progressing DDD prior to the development of glomerulosclerosis. Our data provide evidence supporting the pivotal role of complement alternative pathway

  13. A Software Architecture for High Level Applications

    SciTech Connect

    Shen,G.

    2009-05-04

    A modular software platform for high level applications is under development at the National Synchrotron Light Source II project. This platform is based on client-server architecture, and the components of high level applications on this platform will be modular and distributed, and therefore reusable. An online model server is indispensable for model based control. Different accelerator facilities have different requirements for the online simulation. To supply various accelerator simulators, a set of narrow and general application programming interfaces is developed based on Tracy-3 and Elegant. This paper describes the system architecture for the modular high level applications, the design of narrow and general application programming interface for an online model server, and the prototype of online model server.

  14. Case hardenability at high carbon levels

    SciTech Connect

    Walton, H.W.

    1995-02-01

    Loss of hardenability in the case was thought to be responsible for a lower than specified hardness found on a large carburized bushing. Pseudo Jominy testing on several high hardenability carburizing grades confirmed that hardenability fade was present at carbon levels above 0.65% and particularly for those steels containing molybdenum. Analysis of previous work provided a formula for calculating Jominy hardenability at various carbon levels. Again the results confirmed that the loss of hardenability was more severe in steels containing molybdenum.

  15. Future high sea levels in south Sweden

    SciTech Connect

    Blomgren, S.H.; Hanson, H.

    1997-12-31

    An estimation of future mean high water levels in Oeresund and the southwest Baltic Sea is presented together with a discussion of probable consequences for Falsterbo Peninsula, a trumpet-shaped sandy formation of some 25 km{sup 2} size situated in the very southwest corner of Sweden. A literature review coupled with sea-level measurements and observations made in the area every four hours since October 1945 are given and comprise the base for the present analysis.

  16. Antibodies to gp41 and nef in otherwise HIV-negative homosexual man with Kaposi's sarcoma.

    PubMed

    Bowden, F J; McPhee, D A; Deacon, N J; Cumming, S A; Doherty, R R; Sonza, S; Lucas, C R; Crowe, S M

    1991-06-01

    A homosexual man with histologically confirmed Kaposi's sarcoma remained seronegative for HIV-1, HIV-2, and HTLV-1 on conventional tests over a 4-year period. HIV cultures were also negative on thirteen separate occasions. However, serum antibodies to synthetic peptide analogues of the gp41 and nef regions of HIV-1 were consistently detected on an enzyme immunoassay. Tests with the polymerase chain reaction with primers directed to the gag and env regions were negative. The antigens to which the antibodies were produced might have come from a defective HIV mutant, another retrovirus, or a hitherto unknown "agent of Kaposi's sarcoma" with similar antigenic epitopes.

  17. High-level radioactive wastes. Supplement 1

    SciTech Connect

    McLaren, L.H.

    1984-09-01

    This bibliography contains information on high-level radioactive wastes included in the Department of Energy's Energy Data Base from August 1982 through December 1983. These citations are to research reports, journal articles, books, patents, theses, and conference papers from worldwide sources. Five indexes, each preceded by a brief description, are provided: Corporate Author, Personal Author, Subject, Contract Number, and Report Number. 1452 citations.

  18. PAIRWISE BLENDING OF HIGH LEVEL WASTE (HLW)

    SciTech Connect

    CERTA, P.J.

    2006-02-22

    The primary objective of this study is to demonstrate a mission scenario that uses pairwise and incidental blending of high level waste (HLW) to reduce the total mass of HLW glass. Secondary objectives include understanding how recent refinements to the tank waste inventory and solubility assumptions affect the mass of HLW glass and how logistical constraints may affect the efficacy of HLW blending.

  19. Do we understand high-level vision?

    PubMed

    Cox, David Daniel

    2014-04-01

    'High-level' vision lacks a single, agreed upon definition, but it might usefully be defined as those stages of visual processing that transition from analyzing local image structure to analyzing structure of the external world that produced those images. Much work in the last several decades has focused on object recognition as a framing problem for the study of high-level visual cortex, and much progress has been made in this direction. This approach presumes that the operational goal of the visual system is to read-out the identity of an object (or objects) in a scene, in spite of variation in the position, size, lighting and the presence of other nearby objects. However, while object recognition as a operational framing of high-level is intuitive appealing, it is by no means the only task that visual cortex might do, and the study of object recognition is beset by challenges in building stimulus sets that adequately sample the infinite space of possible stimuli. Here I review the successes and limitations of this work, and ask whether we should reframe our approaches to understanding high-level vision. Copyright © 2014. Published by Elsevier Ltd.

  20. Python based high-level synthesis compiler

    NASA Astrophysics Data System (ADS)

    Cieszewski, Radosław; Pozniak, Krzysztof; Romaniuk, Ryszard

    2014-11-01

    This paper presents a python based High-Level synthesis (HLS) compiler. The compiler interprets an algorithmic description of a desired behavior written in Python and map it to VHDL. FPGA combines many benefits of both software and ASIC implementations. Like software, the mapped circuit is flexible, and can be reconfigured over the lifetime of the system. FPGAs therefore have the potential to achieve far greater performance than software as a result of bypassing the fetch-decode-execute operations of traditional processors, and possibly exploiting a greater level of parallelism. Creating parallel programs implemented in FPGAs is not trivial. This article describes design, implementation and first results of created Python based compiler.

  1. DUACS: Toward High Resolution Sea Level Products

    NASA Astrophysics Data System (ADS)

    Faugere, Y.; Gerald, D.; Ubelmann, C.; Claire, D.; Pujol, M. I.; Antoine, D.; Desjonqueres, J. D.; Picot, N.

    2016-12-01

    The DUACS system produces, as part of the CNES/SALP project, and the Copernicus Marine Environment and Monitoring Service, high quality multimission altimetry Sea Level products for oceanographic applications, climate forecasting centers, geophysic and biology communities... These products consist in directly usable and easy to manipulate Level 3 (along-track cross-calibrated SLA) and Level 4 products (multiple sensors merged as maps or time series) and are available in global and regional version (Mediterranean Sea, Arctic, European Shelves …).The quality of the products is today limited by the altimeter technology "Low Resolution Mode" (LRM), and the lack of available observations. The launch of 2 new satellites in 2016, Jason-3 and Sentinel-3A, opens new perspectives. Using the global Synthetic Aperture Radar mode (SARM) coverage of S3A and optimizing the LRM altimeter processing (retracking, editing, ...) will allow us to fully exploit the fine-scale content of the altimetric missions. Thanks to this increase of real time altimetry observations we will also be able to improve Level-4 products by combining these new Level-3 products and new mapping methodology, such as dynamic interpolation. Finally these improvements will benefit to downstream products : geostrophic currents, Lagrangian products, eddy atlas… Overcoming all these challenges will provide major upgrades of Sea Level products to better fulfill user needs.

  2. High-Level Waste Melter Study Report

    SciTech Connect

    Perez, Joseph M.; Bickford, Dennis F.; Day, Delbert E.; Kim, Dong-Sang; Lambert, Steven L.; Marra, Sharon L.; Peeler, David K.; Strachan, Denis M.; Triplett, Mark B.; Vienna, John D.; Wittman, Richard S.

    2001-07-13

    At the Hanford Site in Richland, Washington, the path to site cleanup involves vitrification of the majority of the wastes that currently reside in large underground tanks. A Joule-heated glass melter is the equipment of choice for vitrifying the high-level fraction of these wastes. Even though this technology has general national and international acceptance, opportunities may exist to improve or change the technology to reduce the enormous cost of accomplishing the mission of site cleanup. Consequently, the U.S. Department of Energy requested the staff of the Tanks Focus Area to review immobilization technologies, waste forms, and modifications to requirements for solidification of the high-level waste fraction at Hanford to determine what aspects could affect cost reductions with reasonable long-term risk. The results of this study are summarized in this report.

  3. High-Level Waste Melter Study Report

    SciTech Connect

    Perez Jr, Joseph M; Bickford, Dennis F; Day, Delbert E; Kim, Dong-Sang; Lambert, Steven L; Marra, Sharon L; Peeler, David K; Strachan, Denis M; Triplett, Mark B; Vienna, John D; Wittman, Richard S

    2001-07-13

    At the Hanford Site in Richland, Washington, the path to site cleanup involves vitrification of the majority of the wastes that currently reside in large underground tanks. A Joule-heated glass melter is the equipment of choice for vitrifying the high-level fraction of these wastes. Even though this technology has general national and international acceptance, opportunities may exist to improve or change the technology to reduce the enormous cost of accomplishing the mission of site cleanup. Consequently, the U.S. Department of Energy requested the staff of the Tanks Focus Area to review immobilization technologies, waste forms, and modifications to requirements for solidification of the high-level waste fraction at Hanford to determine what aspects could affect cost reductions with reasonable long-term risk. The results of this study are summarized in this report.

  4. The HIV-1 Nef protein interacts with two components of the 40S small ribosomal subunit, the RPS10 protein and the 18S rRNA.

    PubMed

    Abbas, Wasim; Dichamp, Isabelle; Herbein, Georges

    2012-07-10

    Human immunodeficiency virus type 1 (HIV-1) Nef-encoded protein plays key functions at almost all stages of the viral life cycle, but its role in translation is largely unknown. To determine the effect of Nef on translation we used an in vitro translation assay. The detection of Nef/RPS10 complexes and the presence of 18S rRNA and tRNAs in the complexes were performed by coimmunoprecipitation and RT-PCR assay. We observed that the HIV-1 Nef protein specifically impaired translation in vitro. We observed the interaction of Nef with RPS10 by coimmunoprecipitation assay. In addition 18S rRNA and tRNAs were present in the Nef/RPS10 complexes. Our results are consistent with a model in which the Nef protein by binding to two components of the 40S small ribosomal subunit, RPS10 and 18S rRNA, and to a lesser extent to tRNAs, could lead to decreased protein synthesis.

  5. Commissioning of the CMS High Level Trigger

    SciTech Connect

    Agostino, Lorenzo; et al.

    2009-08-01

    The CMS experiment will collect data from the proton-proton collisions delivered by the Large Hadron Collider (LHC) at a centre-of-mass energy up to 14 TeV. The CMS trigger system is designed to cope with unprecedented luminosities and LHC bunch-crossing rates up to 40 MHz. The unique CMS trigger architecture only employs two trigger levels. The Level-1 trigger is implemented using custom electronics, while the High Level Trigger (HLT) is based on software algorithms running on a large cluster of commercial processors, the Event Filter Farm. We present the major functionalities of the CMS High Level Trigger system as of the starting of LHC beams operations in September 2008. The validation of the HLT system in the online environment with Monte Carlo simulated data and its commissioning during cosmic rays data taking campaigns are discussed in detail. We conclude with the description of the HLT operations with the first circulating LHC beams before the incident occurred the 19th September 2008.

  6. Genetic drift can dominate short-term human immunodeficiency virus type 1 nef quasispecies evolution in vivo.

    PubMed Central

    Plikat, U; Nieselt-Struwe, K; Meyerhans, A

    1997-01-01

    The evolution of human immunodeficiency virus (HIV) type 1 nef quasispecies in a patient clonally infected with a contaminated batch of blood clotting factor IX was monitored. nef sequences were derived at 11, 25, and 41 months postinfection from infected peripheral blood mononuclear cells after molecular cloning of PCR-amplified proviral DNA. The phylogenetic relationships among a total of 41 informative sequences were established by split decomposition analysis and used as a basis to establish a substitution matrix and to score synonymous (s) and nonsynonymous (ns) substitutions. The number of observed in-phase stop codons within the nef sequences was comparable to that expected on a random basis. Similarly, the numbers of observed s and ns substitutions did not differ significantly from expected values. No codon position was preferentially mutated. The maximum sequence divergence increased in a linear manner, with approximately 4.4 nucleotide and approximately 3.2 amino acid changes per year. It appears that stochastic processes strongly influence short-term HIV nef quasispecies evolution in vivo. PMID:9151810

  7. Performance of the CMS High Level Trigger

    NASA Astrophysics Data System (ADS)

    Perrotta, Andrea

    2015-12-01

    The CMS experiment has been designed with a 2-level trigger system. The first level is implemented using custom-designed electronics. The second level is the so-called High Level Trigger (HLT), a streamlined version of the CMS offline reconstruction software running on a computer farm. For Run II of the Large Hadron Collider, the increases in center-of-mass energy and luminosity will raise the event rate to a level challenging for the HLT algorithms. The increase in the number of interactions per bunch crossing, on average 25 in 2012, and expected to be around 40 in Run II, will be an additional complication. We present here the expected performance of the main triggers that will be used during the 2015 data taking campaign, paying particular attention to the new approaches that have been developed to cope with the challenges of the new run. This includes improvements in HLT electron and photon reconstruction as well as better performing muon triggers. We will also present the performance of the improved tracking and vertexing algorithms, discussing their impact on the b-tagging performance as well as on the jet and missing energy reconstruction.

  8. Liquid level measurement in high level nuclear waste slurries

    SciTech Connect

    Weeks, G.E.; Heckendorn, F.M.; Postles, R.L.

    1990-01-01

    Accurate liquid level measurement has been a difficult problem to solve for the Defense Waste Processing Facility (DWPF). The nuclear waste sludge tends to plug or degrade most commercially available liquid-level measurement sensors. A liquid-level measurement system that meets demanding accuracy requirements for the DWPF has been developed. The system uses a pneumatic 1:1 pressure repeater as a sensor and a computerized error correction system. 2 figs.

  9. Proton Affinity Calculations with High Level Methods.

    PubMed

    Kolboe, Stein

    2014-08-12

    Proton affinities, stretching from small reference compounds, up to the methylbenzenes and naphthalene and anthracene, have been calculated with high accuracy computational methods, viz. W1BD, G4, G3B3, CBS-QB3, and M06-2X. Computed and the currently accepted reference proton affinities are generally in excellent accord, but there are deviations. The literature value for propene appears to be 6-7 kJ/mol too high. Reported proton affinities for the methylbenzenes seem 4-5 kJ/mol too high. G4 and G3 computations generally give results in good accord with the high level W1BD. Proton affinity values computed with the CBS-QB3 scheme are too low, and the error increases with increasing molecule size, reaching nearly 10 kJ/mol for the xylenes. The functional M06-2X fails markedly for some of the small reference compounds, in particular, for CO and ketene, but calculates methylbenzene proton affinities with high accuracy.

  10. The high-level trigger of ALICE

    NASA Astrophysics Data System (ADS)

    Tilsner, H.; Alt, T.; Aurbakken, K.; Grastveit, G.; Helstrup, H.; Lindenstruth, V.; Loizides, C.; Nystrand, J.; Roehrich, D.; Skaali, B.; Steinbeck, T.; Ullaland, K.; Vestbo, A.; Vik, T.

    One of the main tracking detectors of the forthcoming ALICE Experiment at the LHC is a cylindrical Time Projection Chamber (TPC) with an expected data volume of about 75 MByte per event. This data volume, in combination with the presumed maximum bandwidth of 1.2 GByte/s to the mass storage system, would limit the maximum event rate to 20 Hz. In order to achieve higher event rates, online data processing has to be applied. This implies either the detection and read-out of only those events which contain interesting physical signatures or an efficient compression of the data by modeling techniques. In order to cope with the anticipated data rate, massive parallel computing power is required. It will be provided in form of a clustered farm of SMP-nodes, based on off-the-shelf PCs, which are connected with a high bandwidth low overhead network. This High-Level Trigger (HLT) will be able to process a data rate of 25 GByte/s online. The front-end electronics of the individual sub-detectors is connected to the HLT via an optical link and a custom PCI card which is mounted in the clustered PCs. The PCI card is equipped with an FPGA necessary for the implementation of the PCI-bus protocol. Therefore, this FPGA can also be used to assist the host processor with first-level processing. The first-level processing done on the FPGA includes conventional cluster-finding for low multiplicity events and local track finding based on the Hough Transformation of the raw data for high multiplicity events. PACS: 07.05.-t Computers in experimental physics - 07.05.Hd Data acquisition: hardware and software - 29.85.+c Computer data analysis

  11. EAP high-level product architecture

    NASA Astrophysics Data System (ADS)

    Gudlaugsson, T. V.; Mortensen, N. H.; Sarban, R.

    2013-04-01

    EAP technology has the potential to be used in a wide range of applications. This poses the challenge to the EAP component manufacturers to develop components for a wide variety of products. Danfoss Polypower A/S is developing an EAP technology platform, which can form the basis for a variety of EAP technology products while keeping complexity under control. High level product architecture has been developed for the mechanical part of EAP transducers, as the foundation for platform development. A generic description of an EAP transducer forms the core of the high level product architecture. This description breaks down the EAP transducer into organs that perform the functions that may be present in an EAP transducer. A physical instance of an EAP transducer contains a combination of the organs needed to fulfill the task of actuator, sensor, and generation. Alternative principles for each organ allow the function of the EAP transducers to be changed, by basing the EAP transducers on a different combination of organ alternatives. A model providing an overview of the high level product architecture has been developed to support daily development and cooperation across development teams. The platform approach has resulted in the first version of an EAP technology platform, on which multiple EAP products can be based. The contents of the platform have been the result of multi-disciplinary development work at Danfoss PolyPower, as well as collaboration with potential customers and research institutions. Initial results from applying the platform on demonstrator design for potential applications are promising. The scope of the article does not include technical details.

  12. High-level waste qualification: Managing uncertainty

    SciTech Connect

    Pulsipher, B.A.

    1993-09-01

    A vitrification facility is being developed by the U.S. Department of Energy (DOE) at the West Valley Demonstration Plant (WVDP) near Buffalo, New York, where approximately 300 canisters of high-level nuclear waste glass will be produced. To assure that the produced waste form is acceptable, uncertainty must be managed. Statistical issues arise due to sampling, waste variations, processing uncertainties, and analytical variations. This paper presents elements of a strategy to characterize and manage the uncertainties associated with demonstrating that an acceptable waste form product is achieved. Specific examples are provided within the context of statistical work performed by Pacific Northwest Laboratory (PNL).

  13. Regulation of high density lipoprotein levels

    SciTech Connect

    Krauss, R.M.

    1982-03-01

    An increasing awareness of the physiologic and pathologic importance of serum high density lipoproteins (HDL) has led to a large number of observations regarding factors which influence their concentrations. HDL consists of a heterogeneous collection of macromolecules with diverse physical properties and chemical constituents. While laboratory techniques have made it possible to measure HDL and their individual components, there are as yet large gaps in our knowledge of the biochemical mechanisms and clinical significance of changes in these laboratory parameters. In this review, current concepts of the structure and metabolism of HDL will be briefly summarized, and the factors influencing their levels in humans will be surveyed. 313 references.

  14. The effects of high level infrasound

    SciTech Connect

    Johnson, D.L.

    1980-02-01

    This paper will attempt to survey the current knowledge on the effects of relative high levels of infrasound on humans. While this conference is concerned mainly about hearing, some discussion of other physiological effects is appropriate. Such discussion also serves to highlight a basic question, 'Is hearing the main concern of infrasound and low frequency exposure, or is there a more sensitive mechanism'. It would be comforting to know that the focal point of this conference is indeed the most important concern. Therefore, besides hearing loss and auditory threshold of infrasonic and low frequency exposure, four other effects will be provided. These are performance, respiration, annoyance, and vibration.

  15. Service Oriented Architecture for High Level Applications

    SciTech Connect

    Chu, Chungming; Chevtsov, Sergei; Wu, Juhao; Shen, Guobao; /Brookhaven

    2012-06-28

    Standalone high level applications often suffer from poor performance and reliability due to lengthy initialization, heavy computation and rapid graphical update. Service-oriented architecture (SOA) is trying to separate the initialization and computation from applications and to distribute such work to various service providers. Heavy computation such as beam tracking will be done periodically on a dedicated server and data will be available to client applications at all time. Industrial standard service architecture can help to improve the performance, reliability and maintainability of the service. Robustness will also be improved by reducing the complexity of individual client applications.

  16. The High Level Data Reduction Library

    NASA Astrophysics Data System (ADS)

    Ballester, P.; Gabasch, A.; Jung, Y.; Modigliani, A.; Taylor, J.; Coccato, L.; Freudling, W.; Neeser, M.; Marchetti, E.

    2015-09-01

    The European Southern Observatory (ESO) provides pipelines to reduce data for most of the instruments at its Very Large telescope (VLT). These pipelines are written as part of the development of VLT instruments, and are used both in the ESO's operational environment and by science users who receive VLT data. All the pipelines are highly specific geared toward instruments. However, experience showed that the independently developed pipelines include significant overlap, duplication and slight variations of similar algorithms. In order to reduce the cost of development, verification and maintenance of ESO pipelines, and at the same time improve the scientific quality of pipelines data products, ESO decided to develop a limited set of versatile high-level scientific functions that are to be used in all future pipelines. The routines are provided by the High-level Data Reduction Library (HDRL). To reach this goal, we first compare several candidate algorithms and verify them during a prototype phase using data sets from several instruments. Once the best algorithm and error model have been chosen, we start a design and implementation phase. The coding of HDRL is done in plain C and using the Common Pipeline Library (CPL) functionality. HDRL adopts consistent function naming conventions and a well defined API to minimise future maintenance costs, implements error propagation, uses pixel quality information, employs OpenMP to take advantage of multi-core processors, and is verified with extensive unit and regression tests. This poster describes the status of the project and the lesson learned during the development of reusable code implementing algorithms of high scientific quality.

  17. Exposure to unusually high indoor radon levels

    SciTech Connect

    Rasheed, F.N. )

    1993-03-27

    Unusually high indoor radon concentrations were reported in a small village in western Tyrol, Austria. The authors have measured the seasonal course of indoor radon concentrations in 390 houses of this village. 71% of houses in winter and 33% in summer, showed radon values on the ground floor above the Austrian action level of 400 Bq/cm[sup 3]. This proportion results in an unusually high indoor radon exposure of the population. The radon source was an 8,700-year-old rock slide of granite gneiss, the largest of the alpine crystalline rocks. It has a strong emanating power because its rocks are heavily fractured and show a slightly increased uranium content. Previous reports show increased lung cancer mortality, myeloid leukemia, kidney cancer, melanoma, and prostate cancer resulting from indoor radon exposure. However, many studies fail to provide accurate information on indoor radon concentrations, classifying them merely as low, intermediate, and high, or they record only minor increases in indoor radon concentrations. Mortality data for 1970-91 were used to calculate age and sex standardized mortality rates (SMR) for 51 sites of carcinoma. The total population of Tyrol were controls. A significantly higher risk was recorded for lung cancer. The high SMR for lung cancer in female subjects is especially striking. Because the numbers were low for the other cancer sites, these were combined in one group to calculate the SMR. No significant increase in SMR was found for this group.

  18. The ALICE electromagnetic calorimeter high level triggers

    NASA Astrophysics Data System (ADS)

    Ronchetti, F.; Blanco, F.; Figueredo, M.; Knospe, A. G.; Xaplanteris, L.

    2012-12-01

    The ALICE (A Large Ion Collider Experiment) detector yields a huge sample of data from different sub-detectors. On-line data processing is applied to select and reduce the volume of the stored data. ALICE applies a multi-level hardware trigger scheme where fast detectors are used to feed a three-level (L0, L1, and L2) deep chain. The High-Level Trigger (HLT) is a fourth filtering stage sitting logically between the L2 trigger and the data acquisition event building. The EMCal detector comprises a large area electromagnetic calorimeter that extends the momentum measurement of photons and neutral mesons up to pT = 250 GeV/c, which improves the ALICE capability to perform jet reconstruction with measurement of the neutral energy component of jets. An online reconstruction and trigger chain has been developed within the HLT framework to sharpen the EMCal hardware triggers, by combining the central barrel tracking information with the shower reconstruction (clusters) in the calorimeter. In the present report the status and the functionality of the software components developed for the EMCal HLT online reconstruction and trigger chain will be discussed, as well as preliminary results from their commissioning performed during the 2011 LHC running period.

  19. High Level Waste Disposal System Optimization

    SciTech Connect

    Dirk Gombert; M. Connolly; J. Roach; W. Holtzscheiter

    2005-02-01

    The high level waste (HLW) disposal system consists of the Yucca Mountain Facility (YMF) and waste product (e.g. glass) generation facilities. Responsibility for management is shared between the U. S. Department of Energy (DOE) Offices of Civilian Radioactive Waste Management (DOE-RW) and Environmental Management (DOE-EM). The DOE-RW license application and the Waste Acceptance System Requirements Document (WASRD), as well as the DOE-EM Waste Acceptance Product Specification for Vitrified High Level Waste Forms (WAPS) govern the overall performance of the system. This basis for HLW disposal should be reassessed to consider waste form and process technology research and development (R&D), which have been conducted by DOE-EM, international agencies (i.e. ANSTO, CEA), and the private sector; as well as the technical bases for including additional waste forms in the final license application. This will yield a more optimized HLW disposal system to accelerate HLW disposition, more efficient utilization of the YMF, and overall system cost reduction.

  20. High accuracy electronic material level sensor

    DOEpatents

    McEwan, T.E.

    1997-03-11

    The High Accuracy Electronic Material Level Sensor (electronic dipstick) is a sensor based on time domain reflectometry (TDR) of very short electrical pulses. Pulses are propagated along a transmission line or guide wire that is partially immersed in the material being measured; a launcher plate is positioned at the beginning of the guide wire. Reflected pulses are produced at the material interface due to the change in dielectric constant. The time difference of the reflections at the launcher plate and at the material interface are used to determine the material level. Improved performance is obtained by the incorporation of: (1) a high accuracy time base that is referenced to a quartz crystal, (2) an ultrawideband directional sampler to allow operation without an interconnect cable between the electronics module and the guide wire, (3) constant fraction discriminators (CFDs) that allow accurate measurements regardless of material dielectric constants, and reduce or eliminate errors induced by triple-transit or ``ghost`` reflections on the interconnect cable. These improvements make the dipstick accurate to better than 0.1%. 4 figs.

  1. High accuracy electronic material level sensor

    DOEpatents

    McEwan, Thomas E.

    1997-01-01

    The High Accuracy Electronic Material Level Sensor (electronic dipstick) is a sensor based on time domain reflectometry (TDR) of very short electrical pulses. Pulses are propagated along a transmission line or guide wire that is partially immersed in the material being measured; a launcher plate is positioned at the beginning of the guide wire. Reflected pulses are produced at the material interface due to the change in dielectric constant. The time difference of the reflections at the launcher plate and at the material interface are used to determine the material level. Improved performance is obtained by the incorporation of: 1) a high accuracy time base that is referenced to a quartz crystal, 2) an ultrawideband directional sampler to allow operation without an interconnect cable between the electronics module and the guide wire, 3) constant fraction discriminators (CFDs) that allow accurate measurements regardless of material dielectric constants, and reduce or eliminate errors induced by triple-transit or "ghost" reflections on the interconnect cable. These improvements make the dipstick accurate to better than 0.1%.

  2. Mucosal humoral immune response to SIVmac239Δnef vaccination and vaginal challenge

    PubMed Central

    Zeng, Ming; Smith, Anthony J.; Shang, Liang; Wietgrefe, Stephen W.; Voss, James E.; Carlis, John V.; Li, Qingsheng; Piatak, Michael; Lifson, Jeffrey D.; Johnson, R. Paul; Haase, Ashley T.

    2016-01-01

    Live attenuated vaccines such as SIV with a deleted nef gene (SIVmac239Δnef) have provided the most robust protection against subsequent vaginal challenge with wild type (WT) SIV in the SIV-rhesus macaque (RM) model of HIV-1 transmission to women. Hence, identifying correlates of this protection could enable design of an effective HIV-1 vaccine. One such pre-challenge correlate of protection from vaginal challenge has recently been identified as a system with three components: 1) IgG antibodies reacting with the viral envelope glycoprotein, trimeric gp41 (gp41t); 2) produced by plasma cells in the submucosa and ectopic tertiary lymphoid follicles in the ectocervix and vagina; and 3) concentrated on the path of virus entry by the neonatal Fc receptor in the overlying epithelium. We now examine the mucosal production of antibody component of this system post-vaginal challenge. We show that vaginal challenge immediately elicits striking increases in plasma cells not only in the female reproductive tract but also at other mucosal sites, and that these increases correlate with low but persistent replication at mucosal sites. We describe vaginal ectopic follicles that are structurally and functionally organized like follicles in secondary lymphoid organs, and provide inferential evidence for a key role of the female reproductive tract epithelium in facilitating antibody production, affinity maturation and class switch recombination. Vaccination thus accesses an epithelial-immune system axis in the female reproductive tract to respond to exposure to mucosal pathogens. Designing strategies to mimic this system could advance development of an effective HIV-1 vaccine. PMID:26864031

  3. Comparative genetic variability in HIV-1 subtype C nef gene in early age groups of infants.

    PubMed

    Sharma, Uma; Gupta, Poonam; Singhal, Megha; Singh, Supriya; Gupta, Sunil; Venkatesh, Srinivas; Rai, Arvind; Husain, Mohammad

    2017-09-01

    Targeting properties of vertically transmitted viruses in early infancy is important to understand disease progression. To investigate genotypic characteristics of transmitted viruses, blood samples were obtained from infants aged 6 weeks-18 months, categorized in two age groups, acute (<6 months) and early (>6-18 months). Nef having an important role in pathogenesis was selected to explore the viral characteristics. A total of 57 PCR positive samples, amplified by nef gene were sequenced. Analysis showed that 50 sequences belonged to subtype C. In one sequence of acute age group, a long insertion of 10 residues (AAERMRRAEP) in variable region and a 13 residues deletion (ATNNADCAWLEAQ) around proteolytic cleavage region of gene in another sequence was observed. Insertions were also observed in sequences of early age group, however, they ranged from two to eight residues only. In one sequence of early age group, 3/4 arginines at positions 19, 21, 22 of arginine cluster were mutated to glutamine, alanine, and glutamine, respectively. Entropy analysis of two age groups revealed presence of several residues with statistically significant differences in their variability. Among these, 15 (R18,R23,R24; A66,L68,Q71; E74,E77,E78; V87,M92; R119, P144, E167, and C176) belonged to functional motifs, out of which, 12 were in acute age group, suggesting that variability was greater in this group. Prediction of HLA binding peptide motif revealed that epitope LTFGWCFKL was present in >80% study sequences. This epitope was also present in maximum number of HLA types circulating in India and vaccine candidate sequences, suggesting that it may be helpful in designing an epitope-based vaccine. © 2017 Wiley Periodicals, Inc.

  4. Umbra's High Level Architecture (HLA) Interface

    SciTech Connect

    GOTTLIEB, ERIC JOSEPH; MCDONALD, MICHAEL J.; OPPEL III, FRED J.

    2002-04-01

    This report describes Umbra's High Level Architecture HLA library. This library serves as an interface to the Defense Simulation and Modeling Office's (DMSO) Run Time Infrastructure Next Generation Version 1.3 (RTI NG1.3) software library and enables Umbra-based models to be federated into HLA environments. The Umbra library was built to enable the modeling of robots for military and security system concept evaluation. A first application provides component technologies that ideally fit the US Army JPSD's Joint Virtual Battlespace (JVB) simulation framework for Objective Force concept analysis. In addition to describing the Umbra HLA library, the report describes general issues of integrating Umbra with RTI code and outlines ways of building models to support particular HLA simulation frameworks like the JVB.

  5. HIV-1 Nef Binds a Subpopulation of MHC-I throughout Its Trafficking Itinerary and Down-regulates MHC-I by Perturbing Both Anterograde and Retrograde Trafficking*

    PubMed Central

    Yi, Ling; Rosales, Tilman; Rose, Jeremy J.; Chaudhury, Bhabhadeb; Knutson, Jay R.; Venkatesan, Sundararajan

    2010-01-01

    The HIV protein Nef is thought to mediate immune evasion and promote viral persistence in part by down-regulating major histocompatibility complex class I protein (MHC-I or HLA-I) from the cell surface. Two different models have been proposed to explain this phenomenon as follows: 1) stimulation of MHC-I retrograde trafficking from and aberrant recycling to the plasma membrane, and 2) inhibition of anterograde trafficking of newly synthesized HLA-I from the endoplasmic reticulum to the plasma membrane. We show here that Nef simultaneously uses both mechanisms to down-regulate HLA-I in peripheral blood mononuclear cells or HeLa cells. Consistent with this, we found by using fluorescence correlation spectroscopy that a third of diffusing HLA-I at the endoplasmic reticulum, Golgi/trans-Golgi network, and the plasma membrane (PM) was associated with Nef. The binding of Nef was similarly avid for native HLA-I and recombinant HLA-I A2 at the PM. Nef binding to HLA-I at the PM was sensitive to specific inhibition of endocytosis. It was also attenuated by cyclodextrin disruption of PM lipid micro-domain architecture, a change that also retarded lateral diffusion and induced large clusters of HLA-I. In all, our data support a model for Nef down-regulation of HLA-I that involves both major trafficking itineraries and persistent protein-protein interactions throughout the cell. PMID:20622010

  6. Decontamination of high-level waste canisters

    SciTech Connect

    Nesbitt, J.F.; Slate, S.C.; Fetrow, L.K.

    1980-12-01

    This report presents evaluations of several methods for the in-process decontamination of metallic canisters containing any one of a number of solidified high-level waste (HLW) forms. The use of steam-water, steam, abrasive blasting, electropolishing, liquid honing, vibratory finishing and soaking have been tested or evaluated as potential techniques to decontaminate the outer surfaces of HLW canisters. Either these techniques have been tested or available literature has been examined to assess their applicability to the decontamination of HLW canisters. Electropolishing has been found to be the most thorough method to remove radionuclides and other foreign material that may be deposited on or in the outer surface of a canister during any of the HLW processes. Steam or steam-water spraying techniques may be adequate for some applications but fail to remove all contaminated forms that could be present in some of the HLW processes. Liquid honing and abrasive blasting remove contamination and foreign material very quickly and effectively from small areas and components although these blasting techniques tend to disperse the material removed from the cleaned surfaces. Vibratory finishing is very capable of removing the bulk of contamination and foreign matter from a variety of materials. However, special vibratory finishing equipment would have to be designed and adapted for a remote process. Soaking techniques take long periods of time and may not remove all of the smearable contamination. If soaking involves pickling baths that use corrosive agents, these agents may cause erosion of grain boundaries that results in rough surfaces.

  7. HIGH LEVEL RF FOR THE SNS RING.

    SciTech Connect

    ZALTSMAN,A.; BLASKIEWICZ,M.; BRENNAN,J.; BRODOWSKI,J.; METH,M.; SPITZ,R.; SEVERINO,F.

    2002-06-03

    A high level RF system (HLRF) consisting of power amplifiers (PA's) and ferrite loaded cavities is being designed and built by Brookhaven National Laboratory (BNL) for the Spallation Neutron Source (SNS) project. It is a fixed frequency, two harmonic system whose main function is to maintain a gap for the kicker rise time. Three cavities running at the fundamental harmonic (h=l) will provide 40 kV and one cavity at the second harmonic (h=2) will provide 20 kV. Each cavity has two gaps with a design voltage of 10 kV per gap and will be driven by a power amplifier (PA) directly adjacent to it. The PA uses a 600kW tetrode to provide the necessary drive current. The anode of the tetrode is magnetically coupled to the downstream cell of the cavity. Drive to the PA will be provided by a wide band, solid state amplifier located remotely. A dynamic tuning scheme will be implemented to help compensate for the effect of beam loading.

  8. DEFENSE HIGH LEVEL WASTE GLASS DEGRADATION

    SciTech Connect

    W. Ebert

    2001-09-20

    The purpose of this Analysis/Model Report (AMR) is to document the analyses that were done to develop models for radionuclide release from high-level waste (HLW) glass dissolution that can be integrated into performance assessment (PA) calculations conducted to support site recommendation and license application for the Yucca Mountain site. This report was developed in accordance with the ''Technical Work Plan for Waste Form Degradation Process Model Report for SR'' (CRWMS M&O 2000a). It specifically addresses the item, ''Defense High Level Waste Glass Degradation'', of the product technical work plan. The AP-3.15Q Attachment 1 screening criteria determines the importance for its intended use of the HLW glass model derived herein to be in the category ''Other Factors for the Postclosure Safety Case-Waste Form Performance'', and thus indicates that this factor does not contribute significantly to the postclosure safety strategy. Because the release of radionuclides from the glass will depend on the prior dissolution of the glass, the dissolution rate of the glass imposes an upper bound on the radionuclide release rate. The approach taken to provide a bound for the radionuclide release is to develop models that can be used to calculate the dissolution rate of waste glass when contacted by water in the disposal site. The release rate of a particular radionuclide can then be calculated by multiplying the glass dissolution rate by the mass fraction of that radionuclide in the glass and by the surface area of glass contacted by water. The scope includes consideration of the three modes by which water may contact waste glass in the disposal system: contact by humid air, dripping water, and immersion. The models for glass dissolution under these contact modes are all based on the rate expression for aqueous dissolution of borosilicate glasses. The mechanism and rate expression for aqueous dissolution are adequately understood; the analyses in this AMR were conducted to

  9. High bicarbonate levels in narcoleptic children.

    PubMed

    Franco, Patricia; Junqua, Aurelie; Guignard-Perret, Anne; Raoux, Aude; Perier, Magali; Raverot, Veronique; Claustrat, Bruno; Gustin, Marie-Paule; Inocente, Clara Odilia; Lin, Jian-Sheng

    2016-04-01

    The objective of this study was to evaluate the levels of plasma bicarbonate levels in narcoleptic children. Clinical, electrophysiological data and bicarbonate levels were evaluated retrospectively in children seen in our paediatric national reference centre for hypersomnia. The cohort included 23 control subjects (11.5 ± 4 years, 43% boys) and 51 patients presenting de-novo narcolepsy (N) (12.7 ± 3.7 years, 47% boys). In narcoleptic children, cataplexy was present in 78% and DQB1*0602 was positive in 96%. The control children were less obese (2 versus 47%, P = 0.001). Compared with control subjects, narcoleptic children had higher bicarbonate levels (P = 0.02) as well as higher PCO2 (P < 0.01) and lower venous pH gas (P < 0.01). Bicarbonate levels higher than 27 mmol L(-1) were found in 41.2% of the narcoleptic children and 4.2% of the controls (P = 0.001). Bicarbonate levels were correlated with the Adapted Epworth Sleepiness Scale (P = 0.01). Narcoleptic patients without obesity often had bicarbonate levels higher than 27 mmol L (-1) (55 versus 25%, P = 0.025). No differences were found between children with and without cataplexy. In conclusion, narcoleptic patients had higher bicarbonate plasma levels compared to control children. This result could be a marker of hypoventilation in this pathology, provoking an increase in PCO2 and therefore a respiratory acidosis, compensated by an increase in plasma bicarbonates. This simple screening tool could be useful for prioritizing children for sleep laboratory evaluation in practice. © 2015 European Sleep Research Society.

  10. Enhanced Detection of Human Immunodeficiency Virus Type 1 (HIV-1) Nef-Specific T Cells Recognizing Multiple Variants in Early HIV-1 Infection▿ †

    PubMed Central

    Malhotra, Uma; Li, Fusheng; Nolin, Jessica; Allison, Megan; Zhao, Hong; Mullins, James I.; Self, Steve; McElrath, M. Juliana

    2007-01-01

    A human immunodeficiency virus (HIV)-preventive vaccine will likely need to induce broad immunity that can recognize antigens expressed within circulating strains. To understand the potentially relevant responses that T-cell based vaccines should elicit, we examined the ability of T cells from early infected persons to recognize a broad spectrum of potential T-cell epitopes (PTE) expressed by the products encoded by the HIV type 1 (HIV-1) nef gene, which is commonly included in candidate vaccines. T cells were evaluated for gamma interferon (IFN-γ) secretion using two peptide panels: subtype B consensus (CON) peptides and a novel peptide panel providing 70% coverage of PTE in subtype B HIV-1 Nef. Eighteen of 23 subjects’ T cells recognized HIV-1 Nef. In one subject, Nef-specific T cells were detected with the PTE but not with the CON peptides. The greatest frequency of responses spanned Nef amino acids 65 to 103 and 113 to 147, with multiple epitope variants being recognized. Detection of both the epitope domain number and the response magnitude was enhanced using the PTE peptides. On average, we detected 2.7 epitope domains with the PTE peptides versus 1.7 domains with the CON peptides (P = 0.0034). The average response magnitude was 2,169 spot-forming cells (SFC)/106 peripheral blood mononuclear cells (PBMC) with the PTE peptides versus 1,010 SFC/106 PBMC with CON peptides (P = 0.0046). During early HIV-1 infection, Nef-specific T cells capable of recognizing multiple variants are commonly induced, and these responses are readily detected with the PTE peptide panel. Our findings suggest that Nef responses induced by a given vaccine strain before HIV-1 exposure may be sufficiently broad to recognize most variants within subtype B HIV-1. PMID:17329342

  11. Statistics of high-level scene context

    PubMed Central

    Greene, Michelle R.

    2013-01-01

    Context is critical for recognizing environments and for searching for objects within them: contextual associations have been shown to modulate reaction time and object recognition accuracy, as well as influence the distribution of eye movements and patterns of brain activations. However, we have not yet systematically quantified the relationships between objects and their scene environments. Here I seek to fill this gap by providing descriptive statistics of object-scene relationships. A total of 48, 167 objects were hand-labeled in 3499 scenes using the LabelMe tool (Russell et al., 2008). From these data, I computed a variety of descriptive statistics at three different levels of analysis: the ensemble statistics that describe the density and spatial distribution of unnamed “things” in the scene; the bag of words level where scenes are described by the list of objects contained within them; and the structural level where the spatial distribution and relationships between the objects are measured. The utility of each level of description for scene categorization was assessed through the use of linear classifiers, and the plausibility of each level for modeling human scene categorization is discussed. Of the three levels, ensemble statistics were found to be the most informative (per feature), and also best explained human patterns of categorization errors. Although a bag of words classifier had similar performance to human observers, it had a markedly different pattern of errors. However, certain objects are more useful than others, and ceiling classification performance could be achieved using only the 64 most informative objects. As object location tends not to vary as a function of category, structural information provided little additional information. Additionally, these data provide valuable information on natural scene redundancy that can be exploited for machine vision, and can help the visual cognition community to design experiments guided by

  12. Choledocholithiasis presenting with very high transaminase level

    PubMed Central

    Agahi, Amy; McNair, Alistair

    2012-01-01

    We present three cases of choledocholithiasis presenting with a rise in transaminase to levels normally associated with acute hepatitis (alanine aminotransferase in excess of 1000 IU/l). All three cases had repeated investigation for liver disease before identification of common bile duct stones with magnetic resonance cholangiopancreatogram, and removal at endoscopic retrograde cholangiopancreatogram. We discuss the existing literature and the potential mechanisms of hepatocyte injury in extrahepatic obstruction. PMID:23188856

  13. Globalism on the High School Level.

    ERIC Educational Resources Information Center

    Presutti, Robert M.

    1997-01-01

    Describes the International Sibling Program at Lewiston-Porter High School in Youngstown, New York. Notes that 10 "sibling schools" in eight countries participate by exchanging faculty and students. Suggests that the program has given students, staff, and the community many opportunities to interact with the real world. (RS)

  14. High Power Amplifier Harmonic Output Level Measurement

    NASA Technical Reports Server (NTRS)

    Perez, R. M.; Hoppe, D. J.; Khan, A. R.

    1995-01-01

    A method is presented for the measurement of the harmonic output power of high power klystron amplifiers, involving coherent hemispherical radiation pattern measurements of the radiated klystron output. Results are discussed for the operation in saturated and unsaturated conditions, and with a waveguide harmonic filter included.

  15. High Power Amplifier Harmonic Output Level Measurement

    NASA Technical Reports Server (NTRS)

    Perez, R. M.; Hoppe, D. J.; Khan, A. R.

    1995-01-01

    A method is presented for the measurement of the harmonic output power of high power klystron amplifiers, involving coherent hemispherical radiation pattern measurements of the radiated klystron output. Results are discussed for the operation in saturated and unsaturated conditions, and with a waveguide harmonic filter included.

  16. Infrared Thermography in High Level Waste

    SciTech Connect

    GLEATON, DAVIDT.

    2004-08-24

    The Savannah River Site is a Department of Energy, government-owned, company-operated industrial complex built in the 1950s to produce materials used in nuclear weapons. Five reactors were built to support the production of nuclear weapons material. Irradiated materials were moved from the reactors to one of the two chemical separation plants. In these facilities, known as ''canyons,'' the irradiated fuel and target assemblies were chemically processed to separate useful products from waste. Unfortunately, the by-product waste of nuclear material production was a highly radioactive liquid that had to be stored and maintained. In 1993 a strategy was developed to implement predictive maintenance technologies in the Liquid Waste Disposition Project Division responsible for processing the liquid waste. Responsibilities include the processing and treatment of 51 underground tanks designed to hold 750,000 to1,300,000 gallons of liquid waste and operation of a facility that vitrifies highly radioactive liquid waste into glass logs. Electrical and mechanical equipment monitored at these facilities is very similar to that found in non-nuclear industrial plants. Annual inspections are performed on electrical components, roof systems, and mechanical equipment. Troubleshooting and post installation and post-maintenance infrared inspections are performed as needed. In conclusion, regardless of the industry, the use of infrared thermography has proven to be an efficient and effective method of inspection to help improve plant safety and reliability through early detection of equipment problems.

  17. Fermilab Tevatron high level RF accelerating systems

    NASA Astrophysics Data System (ADS)

    Kerns, Q.; Kerns, C.; Miller, H.; Tawser, S.; Reid, J.; Webber, R.; Wildman, D.

    1985-06-01

    Eight tuned RF cavities have been installed and operated in the F0 straight section of the Tevatron. Their mechanical placement along the beam line enables them to be operated for colliding beams as two independent groups of four cavities, group 1-4 accelerating antiprotons and group 5-8 accelerating protons. The only difference is that the spacing between cavities 4 and 5 was increased to stay clear of the F0 colliding point. The cavities can easily be rephased by switching cables in a low-level distribution system (fan-out) so that the full accelerating capability of all eight cavities can be used during a fixed target operations. Likewise, the cables from capacitive probes on each cavity gap can be switched to proper lengths and summed in a fan-back system to give an RF signal representing the amplitude and phase as seen by the beam separately for protons and antiprotons. Such signals have been used to phase lock the Tevatron to the Main Ring for synchronous transfer.

  18. 46 CFR 119.530 - Bilge high level alarms.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Bilge high level alarms. 119.530 Section 119.530... Bilge and Ballast Systems § 119.530 Bilge high level alarms. (a) Each vessel must be provided with a visual and audible alarm at the operating station to indicate a high water level in each of the...

  19. 46 CFR 182.530 - Bilge high level alarms.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Bilge high level alarms. 182.530 Section 182.530... TONS) MACHINERY INSTALLATION Bilge and Ballast Systems § 182.530 Bilge high level alarms. (a) On a... operating station to indicate a high water level in each of the following normally unmanned spaces: (1)...

  20. Bumblebee Pupae Contain High Levels of Aluminium

    PubMed Central

    Exley, Christopher; Rotheray, Ellen; Goulson, David

    2015-01-01

    The causes of declines in bees and other pollinators remains an on-going debate. While recent attention has focussed upon pesticides, other environmental pollutants have largely been ignored. Aluminium is the most significant environmental contaminant of recent times and we speculated that it could be a factor in pollinator decline. Herein we have measured the content of aluminium in bumblebee pupae taken from naturally foraging colonies in the UK. Individual pupae were acid-digested in a microwave oven and their aluminium content determined using transversely heated graphite furnace atomic absorption spectrometry. Pupae were heavily contaminated with aluminium giving values between 13.4 and 193.4 μg/g dry wt. and a mean (SD) value of 51.0 (33.0) μg/g dry wt. for the 72 pupae tested. Mean aluminium content was shown to be a significant negative predictor of average pupal weight in colonies. While no other statistically significant relationships were found relating aluminium to bee or colony health, the actual content of aluminium in pupae are extremely high and demonstrate significant exposure to aluminium. Bees rely heavily on cognitive function and aluminium is a known neurotoxin with links, for example, to Alzheimer’s disease in humans. The significant contamination of bumblebee pupae by aluminium raises the intriguing spectre of cognitive dysfunction playing a role in their population decline. PMID:26042788

  1. Bumblebee pupae contain high levels of aluminium.

    PubMed

    Exley, Christopher; Rotheray, Ellen; Goulson, David

    2015-01-01

    The causes of declines in bees and other pollinators remains an on-going debate. While recent attention has focussed upon pesticides, other environmental pollutants have largely been ignored. Aluminium is the most significant environmental contaminant of recent times and we speculated that it could be a factor in pollinator decline. Herein we have measured the content of aluminium in bumblebee pupae taken from naturally foraging colonies in the UK. Individual pupae were acid-digested in a microwave oven and their aluminium content determined using transversely heated graphite furnace atomic absorption spectrometry. Pupae were heavily contaminated with aluminium giving values between 13.4 and 193.4 μg/g dry wt. and a mean (SD) value of 51.0 (33.0) μg/g dry wt. for the 72 pupae tested. Mean aluminium content was shown to be a significant negative predictor of average pupal weight in colonies. While no other statistically significant relationships were found relating aluminium to bee or colony health, the actual content of aluminium in pupae are extremely high and demonstrate significant exposure to aluminium. Bees rely heavily on cognitive function and aluminium is a known neurotoxin with links, for example, to Alzheimer's disease in humans. The significant contamination of bumblebee pupae by aluminium raises the intriguing spectre of cognitive dysfunction playing a role in their population decline.

  2. Study of the HIV-2 Env Cytoplasmic Tail Variability and Its Impact on Tat, Rev and Nef

    PubMed Central

    Bakouche, Nordine; Vandenbroucke, Anne-Thérèse; Goubau, Patrick; Ruelle, Jean

    2013-01-01

    Background The HIV-2 env’s 3’ end encodes the cytoplasmic tail (CT) of the Env protein. This genomic region also encodes the rev, Tat and Nef protein in overlapping reading frames. We studied the variability in the CT coding region in 46 clinical specimens and in 2 reference strains by sequencing and by culturing. The aims were to analyse the variability of Env CT and the evolution of proteins expressed from overlapping coding sequences. Results A 70% reduction of the length of the CT region affected the HIV-2 ROD and EHO strains in vitro due to a premature stop codon in the env gene. In clinical samples this wasn’t observed, but the CT length varied due to insertions and deletions. We noted 3 conserved and 3 variable regions in the CT. The conserved regions were those containing residues involved in Env endocytosis, the potential HIV-2 CT region implicated in the NF-kB activation and the potential end of the lentiviral lytic peptide one. The variable regions were the potential HIV-2 Kennedy region, the potential lentiviral lytic peptide two and the beginning of the potential lentiviral lytic peptide one. A very hydrophobic region was coded downstream of the premature stop codon observed in vitro, suggesting a membrane spanning region. Interestingly, the nucleotides that are responsible for the variability of the CT don’t impact rev and Nef. However, in the Kennedy-like coding region variability resulted only from nucleotide changes that impacted Env and Tat together. Conclusion The HIV-2 Env, Tat and Rev C-terminal part are subject to major length variations in both clinical samples and cultured strains. The HIV-2 Env CT contains variable and conserved regions. These regions don’t affect the rev and Nef amino acids composition which evolves independently. In contrast, Tat co-evolves with the Env CT. PMID:24223892

  3. Overlapping cis sites used for splicing of HIV-1 env/nef and rev mRNAs.

    PubMed

    Swanson, A K; Stoltzfus, C M

    1998-12-18

    Alternative splicing is used to generate more than 30 human immunodeficiency virus type 1 (HIV-1) spliced and unspliced mRNAs from a single primary transcript. The abundance of HIV-1 mRNAs is determined by the efficiencies with which its different 5' and 3' splice sites are used. Three splice sites (A4c, A4a, and A4b) are upstream of the rev initiator AUG. RNAs spliced at A4c, A4a, and A4b are used as mRNAs for Rev. Another 3' splice site (A5) is immediately downstream of the rev initiator. RNAs spliced at A5 are used as mRNAs for Env and Nef. In this report, primer extension analysis of splicing intermediates was used to show that there are eight branch points in this region, all of which map to adenosine residues. In addition, cis elements recognized by the cellular splicing machinery overlap; the two most 3' branch points overlap with the AG dinucleotides at rev 3' splice sites A4a and A4b. Competition of the overlapping cis sites for different splicing factors may play a role in maintaining the appropriate balance of mRNAs in HIV-1-infected cells. In support of this possibility, mutations at rev 3' splice site A4b AG dinucleotide dramatically increased splicing of the env/nef 3' splice site A5. This correlated with increased usage of the four most 3' branch points, which include those within the rev 3' splice site AG dinucleotides. Consistent with these results, analysis of a mutant in which three of the four env/nef branch points were inactivated indicated that use of splice site A5 was inhibited and splicing was shifted predominantly to the most 5' rev 3' splice site A4c with preferential use of the two most 5' branch points. Our results suggest that spliceosomes formed at rev A4a-4b, rev A4c, and env/nef A5 3' splice sites each recognize different subsets of the eight branch point sequences.

  4. 40 CFR 227.30 - High-level radioactive waste.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 25 2011-07-01 2011-07-01 false High-level radioactive waste. 227.30 Section 227.30 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) OCEAN DUMPING...-level radioactive waste. High-level radioactive waste means the aqueous waste resulting from the...

  5. 40 CFR 227.30 - High-level radioactive waste.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 24 2010-07-01 2010-07-01 false High-level radioactive waste. 227.30 Section 227.30 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) OCEAN DUMPING...-level radioactive waste. High-level radioactive waste means the aqueous waste resulting from the...

  6. 40 CFR 227.30 - High-level radioactive waste.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 26 2013-07-01 2013-07-01 false High-level radioactive waste. 227.30...-level radioactive waste. High-level radioactive waste means the aqueous waste resulting from the operation of the first cycle solvent extraction system, or equivalent, and the concentrated waste...

  7. 40 CFR 227.30 - High-level radioactive waste.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 26 2012-07-01 2011-07-01 true High-level radioactive waste. 227.30...-level radioactive waste. High-level radioactive waste means the aqueous waste resulting from the operation of the first cycle solvent extraction system, or equivalent, and the concentrated waste...

  8. 40 CFR 227.30 - High-level radioactive waste.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 25 2014-07-01 2014-07-01 false High-level radioactive waste. 227.30...-level radioactive waste. High-level radioactive waste means the aqueous waste resulting from the operation of the first cycle solvent extraction system, or equivalent, and the concentrated waste...

  9. Potent, Persistent Induction and Modulation of Cellular Immune Responses in Rhesus Macaques Primed with Ad5hr-Simian Immunodeficiency Virus (SIV) env/rev, gag, and/or nef Vaccines and Boosted with SIV gp120

    PubMed Central

    Patterson, L. Jean; Malkevitch, Nina; Pinczewski, Joel; Venzon, David; Lou, Yuanmei; Peng, Bo; Munch, Cindy; Leonard, Melissa; Richardson, Ersell; Aldrich, Kristine; Kalyanaraman, V. S.; Pavlakis, George N.; Robert-Guroff, Marjorie

    2003-01-01

    Immunity elicited by multicomponent vaccines delivered by replication-competent Ad5hr-simian immunodeficiency virus (SIV) recombinants was systematically investigated. Rhesus macaques were immunized mucosally at weeks 0 and 12 with Ad5hr-SIVsmH4 env/rev, with or without Ad5hr-SIVmac239 gag or Ad5hr-SIVmac239 nef, or with all three recombinants. The total Ad5hr dosage was comparably adjusted among all animals with empty Ad5hr-ΔE3 vector. The macaques were boosted with SIV gp120 in monophosphoryl A-stable emulsion adjuvant at 24 and 36 weeks. Controls received Ad5hr-ΔE3 vector or adjuvant only. By ELISPOT analysis, all four SIV gene products elicited potent cellular immune responses that persisted 42 weeks post-initial immunization. Unexpectedly, modulation of this cellular immune response was observed among macaques receiving one, two, or three Ad5hr-SIV recombinants. Env responses were significantly enhanced throughout the immunization period in macaques immunized with Ad5hr-SIV env/rev plus Ad5hr-SIV gag and tended to be higher in macaques that also received Ad5hr-SIV nef. Macaques primed with all three recombinants displayed significant down-modulation in numbers of gamma interferon (IFN-γ)-secreting cells specific for SIV Nef, and the Env- and Gag-specific responses were also diminished. Modulation of antibody responses was not observed. Down-modulation was seen only during the period of Ad5hr-recombinant priming, not during subunit boosting, although SIV-specific IFN-γ-secreting cells persisted. The effect was not attributable to Ad5hr replication differences among immunization groups. Vaccine delivery via replication-competent live vectors, which can persistently infect new cells and continuously present low-level antigen, may be advantageous in overcoming competition among complex immunogens for immune recognition. Effects of current multicomponent vaccines on individual immune responses should be evaluated with regard to future vaccine design. PMID

  10. Analysis of Cyberbullying Sensitivity Levels of High School Students and Their Perceived Social Support Levels

    ERIC Educational Resources Information Center

    Akturk, Ahmet Oguz

    2015-01-01

    Purpose: The purpose of this paper is to determine the cyberbullying sensitivity levels of high school students and their perceived social supports levels, and analyze the variables that predict cyberbullying sensitivity. In addition, whether cyberbullying sensitivity levels and social support levels differed according to gender was also…

  11. Analysis of Cyberbullying Sensitivity Levels of High School Students and Their Perceived Social Support Levels

    ERIC Educational Resources Information Center

    Akturk, Ahmet Oguz

    2015-01-01

    Purpose: The purpose of this paper is to determine the cyberbullying sensitivity levels of high school students and their perceived social supports levels, and analyze the variables that predict cyberbullying sensitivity. In addition, whether cyberbullying sensitivity levels and social support levels differed according to gender was also…

  12. A Framework for Translating a High Level Security Policy into Low Level Security Mechanisms

    NASA Astrophysics Data System (ADS)

    Hassan, Ahmed A.; Bahgat, Waleed M.

    2010-01-01

    Security policies have different components; firewall, active directory, and IDS are some examples of these components. Enforcement of network security policies to low level security mechanisms faces some essential difficulties. Consistency, verification, and maintenance are the major ones of these difficulties. One approach to overcome these difficulties is to automate the process of translation of high level security policy into low level security mechanisms. This paper introduces a framework of an automation process that translates a high level security policy into low level security mechanisms. The framework is described in terms of three phases; in the first phase all network assets are categorized according to their roles in the network security and relations between them are identified to constitute the network security model. This proposed model is based on organization based access control (OrBAC). However, the proposed model extend the OrBAC model to include not only access control policy but also some other administrative security policies like auditing policy. Besides, the proposed model enables matching of each rule of the high level security policy with the corresponding ones of the low level security policy. Through the second phase of the proposed framework, the high level security policy is mapped into the network security model. The second phase could be considered as a translation of the high level security policy into an intermediate model level. Finally, the intermediate model level is translated automatically into low level security mechanism. The paper illustrates the applicability of proposed approach through an application example.

  13. Distinct transcriptome profiles of Gag-specific CD8+ T cells temporally correlated with the protection elicited by SIVΔnef live attenuated vaccine

    PubMed Central

    Lu, Wuxun; Wan, Yanmin; Ma, Fangrui; Johnson, R. Paul; Li, Qingsheng

    2017-01-01

    The live attenuated vaccine (LAV) SIVmac239Δnef (SIVΔnef) confers the best protection among all the vaccine modalities tested in rhesus macaque model of HIV-1 infection. This vaccine has a unique feature of time-dependent protection: macaques are not protected at 3–5 weeks post vaccination (WPV), whereas immune protection emerges between 15 and 20 WPV. Although the exact mechanisms of the time-dependent protection remain incompletely understood, studies suggested that both cellular and humoral immunities contribute to this time-dependent protection. To further elucidate the mechanisms of protection induced by SIVΔnef, we longitudinally compared the global gene expression profiles of SIV Gag-CM9+ CD8+ (Gag-specific CD8+) T cells from peripheral blood of Mamu-A*01+ rhesus macaques at 3 and 20 WPV using rhesus microarray. We found that gene expression profiles of Gag-specific CD8+ T cells at 20 WPV are qualitatively different from those at 3 WPV. At 20 WPV, the most significant transcriptional changes of Gag-specific CD8+ T cells were genes involved in TCR signaling, differentiation and maturation toward central memory cells, with increased expression of CCR7, TCRα, TCRβ, CD28 and decreased expression of CTLA-4, IFN-γ, RANTES, granzyme A and B. Our study suggests that a higher quality of SIV-specific CD8+ T cells elicited by SIVΔnef over time contributes to the maturation of time-dependent protection. PMID:28333940

  14. Physical Activity Levels in Portuguese High School Physical Education

    ERIC Educational Resources Information Center

    Marmeleira, Jose Francisco Filipe; Aldeias, Nuno Micael Carrasqueira; da Graca, Pedro Miguel dos Santos Medeira

    2012-01-01

    The main aim of this study was to evaluate the physical activity (PA) levels of high school Portuguese students during physical education (PE) and investigate the association of PA levels with students' goal orientation and intrinsic motivation. Forty-six students from three high schools participated. Heart rate telemetry and pedometry were used…

  15. Physical Activity Levels in Portuguese High School Physical Education

    ERIC Educational Resources Information Center

    Marmeleira, Jose Francisco Filipe; Aldeias, Nuno Micael Carrasqueira; da Graca, Pedro Miguel dos Santos Medeira

    2012-01-01

    The main aim of this study was to evaluate the physical activity (PA) levels of high school Portuguese students during physical education (PE) and investigate the association of PA levels with students' goal orientation and intrinsic motivation. Forty-six students from three high schools participated. Heart rate telemetry and pedometry were used…

  16. 46 CFR 182.530 - Bilge high level alarms.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 7 2012-10-01 2012-10-01 false Bilge high level alarms. 182.530 Section 182.530 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) MACHINERY INSTALLATION Bilge and Ballast Systems § 182.530 Bilge high level alarms. (a) On a...

  17. 46 CFR 119.530 - Bilge high level alarms.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false Bilge high level alarms. 119.530 Section 119.530 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE... Bilge and Ballast Systems § 119.530 Bilge high level alarms. (a) Each vessel must be provided with a...

  18. 46 CFR 119.530 - Bilge high level alarms.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Bilge high level alarms. 119.530 Section 119.530 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE... Bilge and Ballast Systems § 119.530 Bilge high level alarms. (a) Each vessel must be provided with a...

  19. 46 CFR 119.530 - Bilge high level alarms.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Bilge high level alarms. 119.530 Section 119.530 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE... Bilge and Ballast Systems § 119.530 Bilge high level alarms. (a) Each vessel must be provided with a...

  20. 46 CFR 182.530 - Bilge high level alarms.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 7 2014-10-01 2014-10-01 false Bilge high level alarms. 182.530 Section 182.530 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) MACHINERY INSTALLATION Bilge and Ballast Systems § 182.530 Bilge high level alarms. (a) On a...

  1. 46 CFR 182.530 - Bilge high level alarms.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 7 2011-10-01 2011-10-01 false Bilge high level alarms. 182.530 Section 182.530 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) MACHINERY INSTALLATION Bilge and Ballast Systems § 182.530 Bilge high level alarms. (a) On a...

  2. Process for solidifying high-level nuclear waste

    DOEpatents

    Ross, Wayne A.

    1978-01-01

    The addition of a small amount of reducing agent to a mixture of a high-level radioactive waste calcine and glass frit before the mixture is melted will produce a more homogeneous glass which is leach-resistant and suitable for long-term storage of high-level radioactive waste products.

  3. Tech Prep Implementation. Self-Assessment Inventory. Middle/High School Level. Postsecondary Level.

    ERIC Educational Resources Information Center

    Partnership for Academic and Career Education, Pendleton, SC.

    These two self-assessment inventories are designed to be used for evaluation of tech prep implementation at the middle/high school and postsecondary levels. The middle/high school version is intended to be completed by the district/school, the postsecondary form by the technical college. The form provides for rating the level of implementation as…

  4. Predictors of Placement in Lower Level versus Higher Level High School Mathematics

    ERIC Educational Resources Information Center

    Archbald, Doug; Farley-Ripple, Elizabeth N.

    2012-01-01

    Educators and researchers have long been interested in determinants of access to honors level and college prep courses in high school. Factors influencing access to upper level mathematics courses are particularly important because of the hierarchical and sequential nature of this subject and because students who finish high school with only lower…

  5. The Presence of a vpu Gene and the Lack of Nef-Mediated Downmodulation of T Cell Receptor-CD3 Are Not Always Linked in Primate Lentiviruses ▿ †

    PubMed Central

    Schmökel, Jan; Sauter, Daniel; Schindler, Michael; Leendertz, Fabian H.; Bailes, Elizabeth; Dazza, Marie-Christine; Saragosti, Sentob; Bibollet-Ruche, Frederic; Peeters, Martine; Hahn, Beatrice H.; Kirchhoff, Frank

    2011-01-01

    Nef is an accessory protein critical for the ability of human and simian immunodeficiency viruses (HIV and SIV) to replicate efficiently in their respective hosts. Previous analyses of members of 15 different primate lentivirus lineages revealed a link between Nef function and the presence of a vpu gene. In particular, Nef proteins of all vpu-containing viruses had lost their ability to downmodulate the T cell (TCR-CD3) receptor. Here we examined Nef proteins from eight additional SIV lineages, including SIVgor, SIVwrc, SIVolc, SIVgri, SIVdrl, SIVlho, SIVden, and SIVasc, from western lowland gorillas, western red colobus monkeys, olive colobus monkeys, grivet monkeys, drills, L'Hoest's monkeys, Dent's mona monkeys, and red-tailed monkeys, respectively. We found that except for the nef gene of SIVdrl, all of them were efficiently expressed and modulated CD4, major histocompatibility complex class I (MHC-I), CD28, CXCR4, and Ii cell surface expression and/or enhanced viral infectivity and replication. Furthermore, the Nef proteins of SIVgri, SIVlho, SIVwrc, SIVolc, and SIVgor antagonized tetherin. As expected, the Nef protein of SIVgor, which carries vpu, failed to downmodulate CD3, whereas those of SIVwrc, SIVgri, SIVlho, and SIVasc, which lack vpu, were capable of performing this function. Surprisingly, however, the Nef protein of the vpu-containing SIVden strain retained the ability to downmodulate TCR-CD3, whereas that of SIVolc, which does not contain vpu, was unable to perform this function. Although the SIVden Vpu is about 20 amino acids shorter than other Vpu proteins, it degrades CD4 and antagonizes tetherin. Our data show that there are exceptions to the link between the presence of a vpu gene and nef alleles deficient in CD3 modulation, indicating that host properties also affect the selective pressure for Nef-mediated disruption of TCR-CD3 signaling. Our results are also further evidence that tetherin antagonism is a common function of primate lentivirus

  6. Pseudo-merohedral Twinning and Noncrystallographic Symmetry in Orthorhombic Crystals of SIVmac239 Nef Core Domain Bound to Different-length TCR Fragments

    SciTech Connect

    Kim, W.; Sigalov, A; Stern, L

    2010-01-01

    HIV/SIV Nef mediates many cellular processes through interactions with various cytoplasmic and membrane-associated host proteins, including the signalling subunit of the T-cell receptor (TCR{zeta}). Here, the crystallization strategy, methods and refinement procedures used to solve the structures of the core domain of the SIVmac239 isolate of Nef (Nef{sub core}) in complex with two different TCR{zeta} fragments are described. The structure of SIVmac239 Nef{sub core} bound to the longer TCR{zeta} polypeptide (Leu51-Asp93) was determined to 3.7 {angstrom} resolution (R{sub work} = 28.7%) in the tetragonal space group P4{sub 3}2{sub 1}2. The structure of SIVmac239 Nef{sub core} in complex with the shorter TCR{zeta} polypeptide (Ala63-Arg80) was determined to 2.05 {angstrom} resolution (R{sub work} = 17.0%), but only after the detection of nearly perfect pseudo-merohedral crystal twinning and proper assignment of the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}. The reduction in crystal space-group symmetry induced by the truncated TCR{zeta} polypeptide appears to be caused by the rearrangement of crystal-contact hydrogen-bonding networks and the substitution of crystallographic symmetry operations by similar noncrystallographic symmetry (NCS) operations. The combination of NCS rotations that were nearly parallel to the twin operation (k, h, -l) and a and b unit-cell parameters that were nearly identical predisposed the P2{sub 1}2{sub 1}2{sub 1} crystal form to pseudo-merohedral twinning.

  7. Stable superconducting magnet. [high current levels below critical temperature

    NASA Technical Reports Server (NTRS)

    Boom, R. W. (Inventor)

    1967-01-01

    Operation of a superconducting magnet is considered. A method is described for; (1) obtaining a relatively high current in a superconducting magnet positioned in a bath of a gas refrigerant; (2) operating a superconducting magnet at a relatively high current level without training; and (3) operating a superconducting magnet containing a plurality of turns of a niobium zirconium wire at a relatively high current level without training.

  8. Reference commercial high-level waste glass and canister definition.

    SciTech Connect

    Slate, S.C.; Ross, W.A.; Partain, W.L.

    1981-09-01

    This report presents technical data and performance characteristics of a high-level waste glass and canister intended for use in the design of a complete waste encapsulation package suitable for disposal in a geologic repository. The borosilicate glass contained in the stainless steel canister represents the probable type of high-level waste product that will be produced in a commercial nuclear-fuel reprocessing plant. Development history is summarized for high-level liquid waste compositions, waste glass composition and characteristics, and canister design. The decay histories of the fission products and actinides (plus daughters) calculated by the ORIGEN-II code are presented.

  9. Reference commercial high-level waste glass and canister definition

    NASA Astrophysics Data System (ADS)

    Slate, S. C.; Ross, W. A.; Partain, W. L.

    1981-09-01

    Technical data and performance characteristics of a high level waste glass and canister intended for use in the design of a complete waste encapsulation package suitable for disposal in a geologic repository are presented. The borosilicate glass contained in the stainless steel canister represents the probable type of high level waste product that is produced in a commercial nuclear-fuel reprocessing plant. Development history is summarized for high level liquid waste compositions, waste glass composition and characteristics, and canister design. The decay histories of the fission products and actinides (plus daughters) calculated by the ORIGEN-II code are presented.

  10. Holism and High Level Wellness in the Treatment of Alcoholism.

    ERIC Educational Resources Information Center

    Bartha, Robert; Davis, Tom

    1982-01-01

    Discusses how a holistic and wellness philosophy is a viable alternative in the treatment of alcoholism. Describes five major dimensions of high-level wellness: nutritional awareness, physical fitness, stress management, environmental sensitivity, and self-responsibility. (RC)

  11. Neptunium estimation in dissolver and high-level-waste solutions

    SciTech Connect

    Pathak, P.N.; Prabhu, D.R.; Kanekar, A.S.; Manchanda, V.K.

    2008-07-01

    This papers deals with the optimization of the experimental conditions for the estimation of {sup 237}Np in spent-fuel dissolver/high-level waste solutions using thenoyltrifluoroacetone as the extractant. (authors)

  12. Glucose level regulation via integral high-order sliding modes.

    PubMed

    Dorel, Lela

    2011-04-01

    Diabetes is a condition in which the body either does not produce enough insulin, or does not properly respond to it. This causes the glucose level in blood to increase. An algorithm based on Integral High-Order Sliding Mode technique is proposed, which keeps the normal blood glucose level automatically releasing insulin into the blood. The system is highly insensitive to inevitable parametric and model uncertainties, measurement noises and small delays.

  13. High-Level Waste System Process Interface Description

    SciTech Connect

    d'Entremont, P.D.

    1999-01-14

    The High-Level Waste System is a set of six different processes interconnected by pipelines. These processes function as one large treatment plant that receives, stores, and treats high-level wastes from various generators at SRS and converts them into forms suitable for final disposal. The three major forms are borosilicate glass, which will be eventually disposed of in a Federal Repository, Saltstone to be buried on site, and treated water effluent that is released to the environment.

  14. Effect of high fluoride and high fat on serum lipid levels and oxidative stress in rabbits.

    PubMed

    Sun, Liyan; Gao, Yanhui; Zhang, Wei; Liu, Hui; Sun, Dianjun

    2014-11-01

    The purpose of this study was to explore the effects of high fluoride and high fat on triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), total antioxidant capacity (T-AOC), lipid peroxide (LPO) and malondialdehyde (MDA) in rabbits. A factorial experimental design was used, with two factors (fluoride and fat) and three levels. Seventy-two male rabbits were randomly assigned into nine groups according to initial weight and serum lipid levels. The rabbits were fed with basic feed, moderate fat feed or high fat feed and drank tap water, fluoridated water at levels of 50 and 100mgfluorion/L freely. Biological materials were collected after 5 months, and serum lipid, T-AOC, LPO, and MDA levels were then measured. Using these data, the separate and interactive effects of high fluoride and high fat were analyzed. High fluoride and high fat both increased serum levels of TC, HDL-C and LDL-C significantly (P<0.05), and there was also a synergistic effect between high fluoride and high fat (P<0.05). High fluoride and high fat had different effects on TG levels: high fat significantly increased TG levels (P<0.01) whereas high fluoride had nothing to do with TG levels (P>0.05). High fat significantly elevated LPO and MDA levels and lowered T-AOC levels in serum (P<0.05). Similarly, high fluoride significantly increased LPO and MDA levels in serum (P<0.05). However, there was no interactive effect between high fat and high fluoride on these indexes. In summary, high fluoride and high fat increased serum TC and LDL-C levels individually and synergistically, and this would cause and aggravate hypercholesterolemia in rabbits. At the same time, high fluoride and high fat both made the accumulation of product of oxidative stress in experimental animals. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Field Trips as Cognitive Motivators for High Level Science Learning

    ERIC Educational Resources Information Center

    Hurley, Marlene M.

    2006-01-01

    Using a composite example of field trips from several years of traveling to Yellowstone with high school biology students, the author illustrates how to raise the cognitive level of science instruction and student learning through science field trips. The author examines what teachers can do to raise the level of both teaching and learning in all…

  16. Field Trips as Cognitive Motivators for High Level Science Learning

    ERIC Educational Resources Information Center

    Hurley, Marlene M.

    2006-01-01

    Using a composite example of field trips from several years of traveling to Yellowstone with high school biology students, the author illustrates how to raise the cognitive level of science instruction and student learning through science field trips. The author examines what teachers can do to raise the level of both teaching and learning in all…

  17. The effect of high altitude on nasal nitric oxide levels.

    PubMed

    Altundag, Aytug; Salihoglu, Murat; Cayonu, Melih; Cingi, Cemal; Tekeli, Hakan; Hummel, Thomas

    2014-09-01

    The aim of the present study was to investigate whether nasal nitric oxide (nNO) levels change in relation to high altitude in a natural setting where the weather conditions were favorable. The present study included 41 healthy volunteers without a history of acute rhinosinusitis within 3 weeks and nasal polyposis. The study group consisted of 31 males (76 %) and 10 females (24 %) and the mean age of the study population was 38 ± 10 years. The volunteers encamped for 2 days in a mountain village at an altitude of 1,500 m above sea level (masl) and proceeded to highlands at an altitude of 2,200 masl throughout the day. The measurements of nNO were done randomly, either first at the mountain village or at sea level. Each participant had nNO values both at sea level and at high altitude at the end of the study. The nNO values of sea level and high altitude were compared to investigate the effect of high altitude on nNO levels. The mean of average nNO measurements at the high altitude was 74.2 ± 41 parts-per-billion (ppb) and the mean of the measurements at sea level was 93.4 ± 45 ppb. The change in nNO depending on the altitude level was statistically significant (p < 0.001). The current investigation showed that nNO levels were decreased at high altitude even if the weather conditions were favorable, such as temperature, humidity, and wind.

  18. High level resistance to aminoglycosides in enterococci from Riyadh.

    PubMed

    Al-Ballaa, S R; Qadri, S M; Al-Ballaa, S R; Kambal, A M; Saldin, H; Al-Qatary, K

    1994-07-01

    Enterococci with high level of aminoglycosides resistance are being reported from different parts of the world with increasing frequency. Treatment of infections caused by such isolates is associated with a high incidence of failure or relapse. This is attributed to the loss of the synergetic effect of aminoglycosides and cell wall active agents against isolates exhibiting this type of resistance. To determine the prevalence of enterococci with high level resistance to aminoglycosides in Riyadh, Saudi Arabia, 241 distinct clinical isolates were examined by disk diffusion method using high content aminoglycosides disks. Seventy-four isolates (30%) were resistant to one or more of the aminoglycosides tested. The most common pattern of resistance was that to streptomycin and kanamycin. Of the 241 isolates tested, 29 (12%) were resistant to high levels of gentamicin, 35 (15%) to tobramycin, 65 (27%) to kanamycin and 53 (22%) to streptomycin. The highest rate of resistance to a high level of gentamicin was found among enterococcal blood isolates (30%). Eighteen of the isolates were identified as Enterococcus faecium, 13 (72%) of these showed high level resistance to two or more of the aminoglycosides tested.

  19. Activity in high-level brain regions reflects visibility of low-level stimuli.

    PubMed

    Imamoglu, F; Heinzle, J; Imfeld, A; Haynes, J-D

    2014-11-15

    Stimulus visibility is associated with neural signals in multiple brain regions, ranging from visual cortex to prefrontal regions. Here we used functional magnetic resonance imaging (fMRI) to investigate to which extent the perceived visibility of a "low-level" grating stimulus is reflected in the brain activity in high-level brain regions. Oriented grating stimuli were presented under varying visibility conditions created by backward masking. Visibility was manipulated using four different stimulus onset asynchronies (SOAs), which created a continuum from invisible to highly visible target stimuli. Brain activity in early visual areas, high-level visual brain regions (fusiform gyrus), as well as parietal and prefrontal brain regions was significantly correlated with subjects' psychometric visibility functions. In addition, increased stimulus visibility was reflected in the functional coupling between low and high-level visual areas. Specifically, neuroimaging signals in the middle occipital gyrus were significantly more correlated with signals in the inferior temporal gyrus when subjects successfully perceived the target stimulus than when they did not. These results provide evidence that not only low-level visual but also high-level brain regions reflect visibility of low-level grating stimuli and that changes in functional connectivity reflect perceived stimulus visibility. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. A Testing Instrument for High School Arabic, Level III.

    ERIC Educational Resources Information Center

    Wolowelsky, Joel B.

    The Arabic language examination was designed for Jewish immigrants from Syria wishing to satisfy New York State language requirements for high school graduation by indicating their proficiency in Arabic. The test is essentially a translation of a state test of Hebrew, and is intended to test Arabic at the third-year high school level. The…

  1. An overview of very high level software design methods

    NASA Technical Reports Server (NTRS)

    Asdjodi, Maryam; Hooper, James W.

    1988-01-01

    Very High Level design methods emphasize automatic transfer of requirements to formal design specifications, and/or may concentrate on automatic transformation of formal design specifications that include some semantic information of the system into machine executable form. Very high level design methods range from general domain independent methods to approaches implementable for specific applications or domains. Applying AI techniques, abstract programming methods, domain heuristics, software engineering tools, library-based programming and other methods different approaches for higher level software design are being developed. Though one finds that a given approach does not always fall exactly in any specific class, this paper provides a classification for very high level design methods including examples for each class. These methods are analyzed and compared based on their basic approaches, strengths and feasibility for future expansion toward automatic development of software systems.

  2. Does high serum uric acid level cause aspirin resistance?

    PubMed

    Yildiz, Bekir S; Ozkan, Emel; Esin, Fatma; Alihanoglu, Yusuf I; Ozkan, Hayrettin; Bilgin, Murat; Kilic, Ismail D; Ergin, Ahmet; Kaftan, Havane A; Evrengul, Harun

    2016-06-01

    In patients with coronary artery disease (CAD), though aspirin inhibits platelet activation and reduces atherothrombotic complications, it does not always sufficiently inhibit platelet function, thereby causing a clinical situation known as aspirin resistance. As hyperuricemia activates platelet turnover, aspirin resistance may be specifically induced by increased serum uric acid (SUA) levels. In this study, we thus investigated the association between SUA level and aspirin resistance in patients with CAD. We analyzed 245 consecutive patients with stable angina pectoris (SAP) who in coronary angiography showed more than 50% occlusion in a major coronary artery. According to aspirin resistance, two groups were formed: the aspirin resistance group (Group 1) and the aspirin-sensitive group (Group 2). Compared with those of Group 2, patients with aspirin resistance exhibited significantly higher white blood cell counts, neutrophil counts, neutrophil-to-lymphocyte ratios, SUA levels, high-sensitivity C-reactive protein levels, and fasting blood glucose levels. After multivariate analysis, a high level of SUA emerged as an independent predictor of aspirin resistance. The receiver-operating characteristic analysis provided a cutoff value of 6.45 mg/dl for SUA to predict aspirin resistance with 79% sensitivity and 65% specificity. Hyperuricemia may cause aspirin resistance in patients with CAD and high SUA levels may indicate aspirin-resistant patients. Such levels should thus recommend avoiding heart attack and stroke by adjusting aspirin dosage.

  3. Human endothelial cells enhance human immunodeficiency virus type 1 replication in CD4+ T cells in a Nef-dependent manner in vitro and in vivo.

    PubMed

    Choi, Jaehyuk; Walker, Jason; Boichuk, Sergei; Kirkiles-Smith, Nancy; Torpey, Nicholas; Pober, Jordan S; Alexander, Louis

    2005-01-01

    Infected CD4+ T cells are the primary sites of human immunodeficiency virus type 1 (HIV-1) replication in vivo. However, signals from professional antigen-presenting cells (APCs), such as dendritic cells and macrophages, greatly enhance HIV-1 replication in T cells. Here, we report that in cocultures, vascular endothelial cells (ECs), which in humans can also serve as APCs, can enhance HIV-1 production of both CCR5- and CXCR4-utilizing strains approximately 50,000-fold. The observed HIV-1 replication enhancement conferred by ECs occurred only in memory CD4+ T cells, required expression of major histocompatibility complex class II (MHC-II) molecules by the ECs, and could not be conferred by fixed ECs, all of which are consistent with a requirement for EC-mediated T-cell activation via T-cell receptor (TCR) signaling. Deletion of nef (Nef-) decreased HIV-1 production by approximately 100-fold in T cells cocultured with ECs but had no effect on virus production in T cells cocultured with professional APCs or fibroblasts induced to express MHC-II. Human ECs do not express B7 costimulators, but Nef- replication in CD4(+)-T-cell and EC cocultures could not be rescued by anti-CD28 antibody. ECs act in trans to enhance wild-type but not Nef- replication and facilitate enhanced wild-type replication in naive T cells when added to T-cell or B-lymphoblastoid cell cocultures, suggesting that ECs also provide a TCR-independent signal to infected T cells. Consistent with these in vitro observations, wild-type HIV-1 replicated 30- to 50-fold more than Nef- in human T cells infiltrating allogeneic human skin grafts on human huPBL-SCID/bg mice, an in vivo model of T-cell activation by ECs. Our studies suggest that ECs, which line the entire cardiovascular system and are, per force, in frequent contact with memory CD4+ T cells, provide signals to HIV-1-infected CD4+ T cells to greatly enhance HIV-1 production in a Nef-dependent manner, a mechanism that could contribute to the

  4. High-level trigger system for the LHC ALICE experiment

    NASA Astrophysics Data System (ADS)

    Bramm, R.; Helstrup, H.; Lien, J.; Lindenstruth, V.; Loizides, C.; Röhrich, D.; Skaali, B.; Steinbeck, T.; Stock, R.; Ullaland, K.; Vestbø, A.; Wiebalck, A.; ALICE Colloboration

    2003-04-01

    The central detectors of the ALICE experiment at LHC will produce a data size of up to 75 MB/ event at an event rate ⩽200 Hz resulting in a data rate of ˜15 GB/ s. Online processing of the data is necessary in order to select interesting (sub)events ("High Level Trigger"), or to compress data efficiently by modeling techniques. Processing this data requires a massive parallel computing system (High Level Trigger System). The system will consist of a farm of clustered SMP-nodes based on off-the-shelf PCs connected with a high bandwidth low latency network.

  5. The effects of high presentation levels on consonant feature transmission

    NASA Astrophysics Data System (ADS)

    Hornsby, Benjamin W. Y.; Trine, Timothy D.; Ohde, Ralph N.

    2005-09-01

    The effect of high speech presentation levels on consonant recognition and feature transmission was assessed in eight participants with normal hearing. Consonant recognition in noise (0 dB signal-to-noise ratio) was measured at five overall speech levels ranging from 65 to 100 dB SPL. Consistent with the work of others, overall percent correct performance decreased as the presentation level of speech increased [e.g., G. A. Studebaker, R. L. Sherbecoe, D. M. McDaniel, and C. A. Gwaltney, J. Acoust. Soc. Am. 105(4), 2431-2444 (1999)]. Confusion matrices were analyzed in terms of relative percent information transmitted at each speech presentation level, as a function of feature. Six feature sets (voicing, place, nasality, duration, frication, and sonorance) were analyzed. Results showed the feature duration (long consonant duration fricatives) to be most affected by increases in level, while the voicing feature was relatively unaffected by increases in level. In addition, alveolar consonants were substantially affected by level, while palatal consonants were not. While the underlying mechanisms responsible for decreases in performance with level increases are unclear, an analysis of common error patterns at high levels suggests that saturation of the neural response and/or a loss of neural synchrony may play a role.

  6. High Frequency Acoustic Propagation using Level Set Methods

    DTIC Science & Technology

    2007-01-01

    solution of the high frequency approximation to the wave equation. Traditional solutions to the Eikonal equation in high frequency acoustics are...curvature can be extracted at any point of the front from the level set function (provided the normal and curvature are well-defined at that point ), and... points per wavelength to resolve the wave). Ray tracing is therefore the current standard for high frequency propagation modeling. LSM may provide

  7. Mercury levels in high-end consumers of fish.

    PubMed

    Hightower, Jane M; Moore, Dan

    2003-04-01

    Consumption of food containing mercury has been identified as a health risk. The U.S. Environmental Protection Agency (U.S. EPA) and the National Academy of Sciences recommend keeping the whole blood mercury level < 5.0 microg/L or the hair level < 1.0 microg/g. This corresponds to a reference dose (RfD) of 0.1 microg/kg body weight per day. All patients in a 1-year period (n = 720) who came for an office visit in a private internal medicine practice in San Francisco, California, were evaluated for mercury excess using the current RfD. One hundred twenty-three patients were tested (93 females, 30 males). Of these, data were statistically analyzed for 89 subjects. Mercury levels ranged from 2.0 to 89.5 microg/L for the 89 subjects. The mean for 66 women was 15 microg/L [standard deviation (SD) = 15], and for 23 men was 13 microg/L (SD = 5); 89% had levels exceeding the RfD. Subjects consumed 30 different forms or types of fish. Swordfish had the highest correlation with mercury level. Sixty-seven patients with serial blood levels over time after stopping fish showed a decline in mercury levels; reduction was significant (p < 0.0001). A substantial fraction of patients had diets high in fish consumption; of these, a high proportion had blood mercury levels exceeding the maximum level recommended by the U.S. EPA and National Academy of Sciences. The mean level for women in this survey was 10 times that of mercury levels found in a recent population survey by the U.S. Centers for Disease Control and Prevention. Some children were > 40 times the national mean.

  8. High-level aminoglycoside resistant enterococci isolated from swine.

    PubMed Central

    Jackson, C. R.; Fedorka-Cray, P. J.; Barrett, J. B.; Ladely, S. R.

    2005-01-01

    Approximately 42% (187/444) of swine enterococci collected between the years 1999 and 2000 exhibited high-level resistance to gentamicin (MIC > or =500 microg/ml), kanamycin (MIC > or =500 microg/ml), or streptomycin (MIC > or =1000 microg/ml). Eight aminoglycoside resistance genes were detected using PCR, most frequently ant(6)-Ia and aac(6')-Ii from Enterococcus faecium. Twenty-four per cent (45/187) of total high-level aminoglycoside-resistant isolates and 26% (4/15) of isolates resistant to high levels of all three antimicrobials were negative for all genes tested. These data suggest that enterococci isolated from swine contain diverse and possibly unidentified aminoglycoside resistance genes. PMID:15816164

  9. High-level aminoglycoside resistant enterococci isolated from swine.

    PubMed

    Jackson, C R; Fedorka-Cray, P J; Barrett, J B; Ladely, S R

    2005-04-01

    Approximately 42% (187/444) of swine enterococci collected between the years 1999 and 2000 exhibited high-level resistance to gentamicin (MIC > or =500 microg/ml), kanamycin (MIC > or =500 microg/ml), or streptomycin (MIC > or =1000 microg/ml). Eight aminoglycoside resistance genes were detected using PCR, most frequently ant(6)-Ia and aac(6')-Ii from Enterococcus faecium. Twenty-four per cent (45/187) of total high-level aminoglycoside-resistant isolates and 26% (4/15) of isolates resistant to high levels of all three antimicrobials were negative for all genes tested. These data suggest that enterococci isolated from swine contain diverse and possibly unidentified aminoglycoside resistance genes.

  10. High Frequency Monitoring System of Groundwater Level in Sheliao

    NASA Astrophysics Data System (ADS)

    Lee, C.; Chia, Y.; Chuang, P.

    2012-12-01

    Long-term groundwater monitoring had been executed since 1950s in Taiwan. In 1980s, with improving technology, various types of automatic reorders of groundwater level had become the most widely used equipment in groundwater monitoring. Among these devices, submersible pressure transducer is frequently selected to monitor groundwater level for its high frequency and high resolution. In this study, it is chosen to monitor groundwater level change in Sheliao well. On the other hand, factors which might influence the performance of recorded data were excluded in the early stage of establishment as well. And the final approach is to achieve a comprehensive understanding of the minor groundwater level change of Sheliao well, and specify its connection between precipitation, atmosphere, earth tide and earthquake. The Shelia well is located in central Taiwan, constructed in an unconfined aquifer, recorded hourly groundwater level change since 1997. We tried to establish a 1 Hz sampling rate pressure-sensing system in 2011 June. The groundwater level was monitored in a resolution of 2-mm. According to the records, several small-scale of fluctuations were observed and were all correlate well to the earthquakes. However, during the time that no earthquake occurred, some short-term fluctuations were still occurred, performed in a different pattern to those induced by earthquakes. After further investigation, those anomalous fluctuations of groundwater level were found corresponded to precipitation quite well. The fluctuations were observed under some specific condition, which involving different range of accumulated precipitation, rainfall intensity, and rainfall duration. The result implied groundwater level in Sheliao well changes with loading effect result from runoff on the ground surface and infiltration. And the earth tide lead to regularly change was also observed. We conclude that Sheliao can be characterized as a partial-confined aquifer with high frequency and high

  11. Photovoltaic (PV) Impact Assessment for Very High Penetration Levels

    SciTech Connect

    Cheng, Danling; Mather, Barry A.; Seguin, Richard; Hambrick, Joshua; Broadwater, Robert P.

    2016-01-01

    This paper describes a granular approach for investigating the impacts of very high photovoltaic (PV) generation penetration. Studies on two real-world distribution feeders connected to PV plants are presented. The studies include both steady-state and time-series power flow analyses, which include the effects of solar variability. The goal of the study is to predict the effects of increasing levels of PV generation as it reaches very high penetration levels. The loss and return of generation with and without regulation is simulated to capture short-term problems such as voltage fluctuations. Impact results from the analyses are described along with potential mitigations.

  12. High level radioactive waste management facility design criteria

    SciTech Connect

    Sheikh, N.A.; Salaymeh, S.R.

    1993-10-01

    This paper discusses the engineering systems for the structural design of the Defense Waste Processing Facility (DWPF) at the Savannah River Site (SRS). At the DWPF, high level radioactive liquids will be mixed with glass particles and heated in a melter. This molten glass will then be poured into stainless steel canisters where it will harden. This process will transform the high level waste into a more stable, manageable substance. This paper discuss the structural design requirements for this unique one of a kind facility. A special emphasis will be concentrated on the design criteria pertaining to earthquake, wind and tornado, and flooding.

  13. Final report on cermet high-level waste forms

    SciTech Connect

    Kobisk, E.H.; Quinby, T.C.; Aaron, W.S.

    1981-08-01

    Cermets are being developed as an alternate method for the fixation of defense and commercial high level radioactive waste in a terminal disposal form. Following initial feasibility assessments of this waste form, consisting of ceramic particles dispersed in an iron-nickel base alloy, significantly improved processing methods were developed. The characterization of cermets has continued through property determinations on samples prepared by various methods from a variety of simulated and actual high-level wastes. This report describes the status of development of the cermet waste form as it has evolved since 1977. 6 tables, 18 figures.

  14. High Level Waste (HLW) Feed Process Control Strategy

    SciTech Connect

    STAEHR, T.W.

    2000-06-14

    The primary purpose of this document is to describe the overall process control strategy for monitoring and controlling the functions associated with the Phase 1B high-level waste feed delivery. This document provides the basis for process monitoring and control functions and requirements needed throughput the double-shell tank system during Phase 1 high-level waste feed delivery. This document is intended to be used by (1) the developers of the future Process Control Plan and (2) the developers of the monitoring and control system.

  15. TE-domestication and horizontal transfer in a putative Nef-AP1mu mimic of HLA-A cytoplasmic domain re-trafficking

    PubMed Central

    Murray, Joseph S.; Murray, Elaina H.

    2016-01-01

    ABSTRACT Genes of the major histocompatibility complex (MHC; also called HLA in human) are polymorphic elements in the genomes of sharks to humans. Class-I and class-II MHC loci appear responsible for much of the genetic linkage to myriad disease states via the capacity to bind short (~8-15 a.a.) peptides of a given pathogen's proteome, or in some cases, the altered proteomes of cancerous cells, and even (in autoimmunity) certain nominal ‘self’ peptides (Janeway, 2004).1 Unfortunately, little is known about how the canonical structure of the MHC-I/-II peptide-presenting gene evolved, particularly since beyond ~500 Mya (sharks) no paralogs exist.2,3 We previously reported that HLA-A isotype alleles with the α1-helix, R65 motif, are wide-spread in phylogeny, but that the α 2-helix, H151R motif, has apparently segregated out of most species. Surprisingly, an uncharacterized orf in T. syrichta (Loc-103275158) encoded R151, but within a truncated A-23 like gene containing 5′- and 3′- footprints of the transposon (TE), tigger-1; the extant tarsier A-23 allele is totally missing exon-3 and part-of exon-4; together, suggesting TE-mediated inactivation of an intact/ancestral A-23 allele (Murray, 2015a).4 The unique Loc-103275158 orf encodes a putative 15-exon transcript with no apparent paralogs throughout phylogeny. However, an HLA-A11 like gene in M. leucophaeus with a shortened C-terminal domain, and an HLA-A like orf in C. atys with two linked α1/α2/α3 domains, both contain a second transmembrane segment, which is conserved in Loc-103275158. Thus, we could model the putative protein with its Nef-like tail domain docked to its MHC-I like α3 domain (i.e., on the same side of a membrane). This modeled tertiary structure is strikingly similar to the solved structure of the Nef:MHC-I CD:AP1mu transporter (Jia, 2012).5 Nef:AP1mu binds the CD of MHC-I in trafficking MHC-I away from the trans-golgi and into the endocytic pathway in HIV-1 infected cells. The CD

  16. TE-domestication and horizontal transfer in a putative Nef-AP1mu mimic of HLA-A cytoplasmic domain re-trafficking.

    PubMed

    Murray, Joseph S; Murray, Elaina H

    2016-01-01

    Genes of the major histocompatibility complex (MHC; also called HLA in human) are polymorphic elements in the genomes of sharks to humans. Class-I and class-II MHC loci appear responsible for much of the genetic linkage to myriad disease states via the capacity to bind short (~8-15 a.a.) peptides of a given pathogen's proteome, or in some cases, the altered proteomes of cancerous cells, and even (in autoimmunity) certain nominal 'self' peptides (Janeway, 2004).(1) Unfortunately, little is known about how the canonical structure of the MHC-I/-II peptide-presenting gene evolved, particularly since beyond ~500 Mya (sharks) no paralogs exist.(2,3) We previously reported that HLA-A isotype alleles with the α1-helix, R65 motif, are wide-spread in phylogeny, but that the α 2-helix, H151R motif, has apparently segregated out of most species. Surprisingly, an uncharacterized orf in T. syrichta (Loc-103275158) encoded R151, but within a truncated A-23 like gene containing 5'- and 3'- footprints of the transposon (TE), tigger-1; the extant tarsier A-23 allele is totally missing exon-3 and part-of exon-4; together, suggesting TE-mediated inactivation of an intact/ancestral A-23 allele (Murray, 2015a).(4) The unique Loc-103275158 orf encodes a putative 15-exon transcript with no apparent paralogs throughout phylogeny. However, an HLA-A11 like gene in M. leucophaeus with a shortened C-terminal domain, and an HLA-A like orf in C. atys with two linked α1/α2/α3 domains, both contain a second transmembrane segment, which is conserved in Loc-103275158. Thus, we could model the putative protein with its Nef-like tail domain docked to its MHC-I like α3 domain (i.e., on the same side of a membrane). This modeled tertiary structure is strikingly similar to the solved structure of the Nef:MHC-I CD:AP1mu transporter (Jia, 2012).(5) Nef:AP1mu binds the CD of MHC-I in trafficking MHC-I away from the trans-golgi and into the endocytic pathway in HIV-1 infected cells. The CD loop of the

  17. Low voltage to high voltage level shifter and related methods

    NASA Technical Reports Server (NTRS)

    Mentze, Erik J. (Inventor); Hess, Herbert L. (Inventor); Buck, Kevin M. (Inventor); Cox, David F. (Inventor)

    2006-01-01

    A shifter circuit comprises a high and low voltage buffer stages and an output buffer stage. The high voltage buffer stage comprises multiple transistors arranged in a transistor stack having a plurality of intermediate nodes connecting individual transistors along the stack. The transistor stack is connected between a voltage level being shifted to and an input voltage. An inverter of this stage comprises multiple inputs and an output. Inverter inputs are connected to a respective intermediate node of the transistor stack. The low voltage buffer stage has an input connected to the input voltage and an output, and is operably connected to the high voltage buffer stage. The low voltage buffer stage is connected between a voltage level being shifted away from and a lower voltage. The output buffer stage is driven by the outputs of the high voltage buffer stage inverter and the low voltage buffer stage.

  18. Transmission Level High Temperature Superconducting Fault Current Limiter

    SciTech Connect

    Stewart, Gary

    2016-10-05

    The primary objective of this project was to demonstrate the feasibility and reliability of utilizing high-temperature superconducting (HTS) materials in a Transmission Level Superconducting Fault Current Limiter (SFCL) application. During the project, the type of high-temperature superconducting material used evolved from 1st generation (1G) BSCCO-2212 melt cast bulk high-temperature superconductors to 2nd generation (2G) YBCO-based high-temperature superconducting tape. The SFCL employed SuperPower's “Matrix” technology, that offers modular features to enable scale up to transmission voltage levels. The SFCL consists of individual modules that contain elements and parallel inductors that assist in carrying the current during the fault. A number of these modules are arranged in an m x n array to form the current-limiting matrix.

  19. Online pattern recognition for the ALICE high level trigger

    NASA Astrophysics Data System (ADS)

    Bramm, R.; Helstrup, H.; Lien, J.; Lindenstruth, V.; Loizides, C.; Rohrich, D.; Skaali, B.; Steinbeck, T.; Stock, R.; Ullaland, K.; Vestbø, A.; Wiebalck, A.; Alice Collaboration

    2003-04-01

    The ALICE High Level Trigger system needs to reconstruct events online at high data rates. Focusing on the Time Projection Chamber we present two pattern recognition methods under investigation: the sequential approach (cluster finding, track follower) and the iterative approach (Hough Transform, cluster assignment, re-fitting). The implementation of the former in hardware indicates that we can reach the designed inspection rate for p-p collisions of 1 kHz with 98% efficiency.

  20. Cake: Enabling High-level SLOs on Shared Storage Systems

    DTIC Science & Technology

    2012-11-07

    Cake : Enabling High-level SLOs on Shared Storage Systems Andrew Wang Shivaram Venkataraman Sara Alspaugh Randy H. Katz Ion Stoica Electrical...control number. 1. REPORT DATE 07 NOV 2012 2. REPORT TYPE 3. DATES COVERED 00-00-2012 to 00-00-2012 4. TITLE AND SUBTITLE Cake : Enabling High...SUPPLEMENTARY NOTES 14. ABSTRACT Cake is a coordinated, multi-resource scheduler for shared distributed storage environments with the goal of

  1. The tracking of high level waste shipments-TRANSCOM system

    SciTech Connect

    Johnson, P.E.; Joy, D.S.; Pope, R.B.

    1995-12-31

    The TRANSCOM (transportation tracking and communication) system is the U.S. Department of Energy`s (DOE`s) real-time system for tracking shipments of spent fuel, high-level wastes, and other high-visibility shipments of radioactive material. The TRANSCOM system has been operational since 1988. The system was used during FY1993 to track almost 100 shipments within the US.DOE complex, and it is accessed weekly by 10 to 20 users.

  2. Evolution of human immunodeficiency virus type 1 nef and long terminal repeat sequences over 4 years in vivo and in vitro.

    PubMed Central

    Delassus, S; Cheynier, R; Wain-Hobson, S

    1991-01-01

    The evolution of an 851-bp segment of the human immunodeficiency virus type 1 (HIV-1) genome encoding the nef open reading frame and U3/R elements of the long terminal repeat has been followed over a 4-year period in vivo and in vitro. The population of viral sequences at any given time was established by sequencing cloned polymerase chain reaction products. The samples studied were derived from the same man for whom a detailed analysis of the tat gene was previously described (A. Meyerhans, R. Cheynier, J. Albert, M. Seth, S. Kwok, J. Sninsky, L. Morfeldt-Manson, B. Asjö, and S. Wain-Hobson, Cell 58:901-910, 1989). Once again in vitro culture resulted in the selection of minor forms. Over a 4-year period in vivo, there was no obvious selection for, or outgrowth of, any particular nef or U3/R sequence. Few defective nef protein sequences were observed, which argues against nef acting as a negative regulatory factor. Although no functionally defective promoter/trans-activation-responsive elements were identified, the transactivation efficiencies varied between 0.2 and 2 times that of the control. The sequence encoding the most efficient trans-activation-responsive region did not outgrow others. The extreme genetic heterogeneity of the different samples of the locus, either in vivo or in vitro, indicates that there is no such thing as a single, distinct HIV sequence. It is suggested that different HIV-1 loci evolve independently, recombination being responsible for their uncoupling. PMID:1985198

  3. Equity-Focused Schools Carry All Students to High Levels

    ERIC Educational Resources Information Center

    WestEd, 2014

    2014-01-01

    Despite decades of experience supporting efforts from local to state levels to improve learning for underserved students, Sonia Caus Gleason and WestEd's Nancy Gerzon could not point to examples of entire schools accomplishing what they believed was possible: high-poverty public schools personalizing learning for all students to consistently reach…

  4. High level cognitive information processing in neural networks

    NASA Technical Reports Server (NTRS)

    Barnden, John A.; Fields, Christopher A.

    1992-01-01

    Two related research efforts were addressed: (1) high-level connectionist cognitive modeling; and (2) local neural circuit modeling. The goals of the first effort were to develop connectionist models of high-level cognitive processes such as problem solving or natural language understanding, and to understand the computational requirements of such models. The goals of the second effort were to develop biologically-realistic model of local neural circuits, and to understand the computational behavior of such models. In keeping with the nature of NASA's Innovative Research Program, all the work conducted under the grant was highly innovative. For instance, the following ideas, all summarized, are contributions to the study of connectionist/neural networks: (1) the temporal-winner-take-all, relative-position encoding, and pattern-similarity association techniques; (2) the importation of logical combinators into connection; (3) the use of analogy-based reasoning as a bridge across the gap between the traditional symbolic paradigm and the connectionist paradigm; and (4) the application of connectionism to the domain of belief representation/reasoning. The work on local neural circuit modeling also departs significantly from the work of related researchers. In particular, its concentration on low-level neural phenomena that could support high-level cognitive processing is unusual within the area of biological local circuit modeling, and also serves to expand the horizons of the artificial neural net field.

  5. High-Level Overview of Data Needs for RE Analysis

    SciTech Connect

    Lopez, Anthony

    2016-12-22

    This presentation provides a high level overview of analysis topics and associated data needs. Types of renewable energy analysis are grouped into two buckets: First, analysis for renewable energy potential, and second, analysis for other goals. Data requirements are similar but and they build upon one another.

  6. High-Level Visual Object Representations Are Constrained by Position

    PubMed Central

    Kriegeskorte, Nikolaus; Baker, Chris I.

    2010-01-01

    It is widely assumed that high-level visual object representations are position-independent (or invariant). While there is sensitivity to position in high-level object-selective cortex, position and object identity are thought to be encoded independently in the population response such that position information is available across objects and object information is available across positions. Contrary to this view, we show, with both behavior and neuroimaging, that visual object representations are position-dependent (tied to limited portions of the visual field). Behaviorally, we show that the effect of priming an object was greatly reduced with any change in position (within- or between-hemifields), indicating nonoverlapping representations of the same object across different positions. Furthermore, using neuroimaging, we show that object-selective cortex is not only highly sensitive to object position but also the ability to differentiate objects based on its response is greatly reduced across different positions, consistent with the observed behavior and the receptive field properties observed in macaque object-selective neurons. Thus, even at the population level, the object information available in response of object-selective cortex is constrained by position. We conclude that even high-level visual object representations are position-dependent. PMID:20351021

  7. High-level visual object representations are constrained by position.

    PubMed

    Kravitz, Dwight J; Kriegeskorte, Nikolaus; Baker, Chris I

    2010-12-01

    It is widely assumed that high-level visual object representations are position-independent (or invariant). While there is sensitivity to position in high-level object-selective cortex, position and object identity are thought to be encoded independently in the population response such that position information is available across objects and object information is available across positions. Contrary to this view, we show, with both behavior and neuroimaging, that visual object representations are position-dependent (tied to limited portions of the visual field). Behaviorally, we show that the effect of priming an object was greatly reduced with any change in position (within- or between-hemifields), indicating nonoverlapping representations of the same object across different positions. Furthermore, using neuroimaging, we show that object-selective cortex is not only highly sensitive to object position but also the ability to differentiate objects based on its response is greatly reduced across different positions, consistent with the observed behavior and the receptive field properties observed in macaque object-selective neurons. Thus, even at the population level, the object information available in response of object-selective cortex is constrained by position. We conclude that even high-level visual object representations are position-dependent.

  8. Rice lines with high leaf mineral nutrient levels

    USDA-ARS?s Scientific Manuscript database

    Phosphorus (P) and Potassium (K), and sometimes other mineral nutrients are often applied as fertilizer, in addition to Nitrogen, to help achieve high yields in Texas rice production. For some mineral nutrients, total levels in soil would be sufficient to support the desired rice crop growth, but th...

  9. High-level manpower movement and Japan's foreign aid.

    PubMed

    Furuya, K

    1992-01-01

    "Japan's technical assistance programs to Asian countries are summarized. Movements of high-level manpower accompanying direct foreign investments by private enterprise are also reviewed. Proposals for increased human resources development include education and training of foreigners in Japan as well as the training of Japanese aid experts and the development of networks for information exchange."

  10. Typewriter Modifications for Persons Who Are High-Level Quadriplegics.

    ERIC Educational Resources Information Center

    O'Reagan, James R.; And Others

    Standard, common electric typewriters are not completely suited to the needs of a high-level quadriplegic typing with a mouthstick. Experiences show that for complete control of a typewriter a mouthstick user needs the combined features of one-button correction, electric forward and reverse indexing, and easy character viewing. To modify a…

  11. Student Achievement Levels Climb at Ribault Senior High School.

    ERIC Educational Resources Information Center

    Profiles, Programs & Products, 1983

    1983-01-01

    Ribault (Florida) Senior High School has reported a dramatic increase in student achievement levels since implementing a comprehensive college preparation curriculum. Among the program changes that contributed to those gains are: (1) the establishment and consistent enforcement of a strong discipline program, including rules for student conduct, a…

  12. Structuring Peer Interaction To Promote High-Level Cognitive Processing.

    ERIC Educational Resources Information Center

    King, Alison

    2002-01-01

    Examines the kind of peer learning that demands high-level cognitive processing, discussing how peer interaction influences cognitive processes (structuring peer interaction and using guided reciprocal peer questioning); how to promote cognitive processing (knowledge construction and integration and socio- cognitive conflict); metacognition; and…

  13. The Estuary Guide. Level 3: High School. Draft.

    ERIC Educational Resources Information Center

    Alexander, Glen; And Others

    Estuaries are marine systems that serve as nurseries for animals, links in the migratory pathways, and habitat for a complex community of organisms. This curriculum guide intended for use at the high school level seeks to teach what estuaries are; provide opportunities to practice decision-making that affects estuaries; and encourage students to…

  14. Typewriter Modifications for Persons Who Are High-Level Quadriplegics.

    ERIC Educational Resources Information Center

    O'Reagan, James R.; And Others

    Standard, common electric typewriters are not completely suited to the needs of a high-level quadriplegic typing with a mouthstick. Experiences show that for complete control of a typewriter a mouthstick user needs the combined features of one-button correction, electric forward and reverse indexing, and easy character viewing. To modify a…

  15. THE XAL INFRASTRUCTURE FOR HIGH LEVEL CONTROL ROOM APPLICATIONS

    SciTech Connect

    Shishlo, Andrei P; Allen, Christopher K; Chu, Paul; Galambos, John D; Pelaia II, Tom

    2009-01-01

    XAL is a Java programming framework for building high-level control applications related to accelerator physics. The structure, details of implementation, and interaction between components, auxiliary XAL packages, and the latest modifications are discussed. A general overview of XAL applications created for the SNS project is presented.

  16. A comparison of high-level waste form characteristics

    SciTech Connect

    Salmon, R.; Notz, K.J.

    1991-01-01

    There are currently about 1055 million curies of high-level waste with a thermal output of about 2950 kilowatts (KW) at four sites in the United States: West Valley Demonstration Project (WVDP), Savannah River Site (SRS), Hanford Site (HANF), and Idaho National Engineering Laboratory (INEL). These quantities are expected to increase to about 1200 million curies and 3570 kw by the end of year 2020. Under the Nuclear Waste Policy Act, this high-level waste must ultimately be disposed of in a geologic repository. Accordingly, canisters of high-level waste immobilized in borosilicate glass or glass-ceramic mixtures are to be produced at the four sites and stored there until a repository becomes available. Data on the estimated production schedules and on the physical, chemical, and radiological characteristics of the canisters of immobilized high-level waste have been collected in OCRWM's Waste Characteristics Data Base, including recent updates an revisions. Comparisons of some of these data for the four sites are presented in this report. 14 refs., 3 tabs.

  17. Deep-level transient conductance spectroscopy of high resistivity semiconductors

    NASA Astrophysics Data System (ADS)

    Alexiev, Dimitri; Prokopovich, Dale; Reinhard, Mark I.; Thomson, Stuart; Mo, Li

    2005-03-01

    We describe a deep-level transient-conductance (DLTC) spectrometer for high resistivity semiconductors, which uses a radiofrequency (40 MHz) marginal oscillator as conductance detector. The DLTC spectra are generated by periodically filling the deep-level trapping centres by carriers stimulated by a pulsed GaAs laser. Then the trap-emptying conductance's signal process through an exponential Miller correlator as the sample temperature is slowly ramped. A simple capacitive coupling of the samples to the oscillator tank circuit eliminates problems such as unwanted defect annealing and other material changes often associated with the high-temperature techniques necessary for ohmic contact formation. Representative deep-level spectra for CdTe, CdZnTe, HgI and gamma-irradiated Si are given.

  18. Differential regulation of NF-κB-mediated proviral and antiviral host gene expression by primate lentiviral Nef and Vpu proteins.

    PubMed

    Sauter, Daniel; Hotter, Dominik; Van Driessche, Benoît; Stürzel, Christina M; Kluge, Silvia F; Wildum, Steffen; Yu, Hangxing; Baumann, Bernd; Wirth, Thomas; Plantier, Jean-Christophe; Leoz, Marie; Hahn, Beatrice H; Van Lint, Carine; Kirchhoff, Frank

    2015-02-03

    NF-κB is essential for effective transcription of primate lentiviral genomes and also activates antiviral host genes. Here, we show that the early protein Nef of most primate lentiviruses enhances NF-κB activation. In contrast, the late protein Vpu of HIV-1 and its simian precursors inhibits activation of NF-κB, even in the presence of Nef. Although this effect of Vpu did not correlate with its ability to interact with β-TrCP, it involved the stabilization of IκB and reduced nuclear translocation of p65. Interestingly, however, Vpu did not affect casein kinase II-mediated phosphorylation of p65. Lack of Vpu was associated with increased NF-κB activation and induction of interferon and interferon-stimulated genes (ISGs) in HIV-1-infected T cells. Thus, HIV-1 and its simian precursors employ Nef to boost NF-κB activation early during the viral life cycle to initiate proviral transcription, while Vpu is used to downmodulate NF-κB-dependent expression of ISGs at later stages. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  19. On Programming Models for Service-Level High Availability

    SciTech Connect

    Engelmann, Christian; Scott, Stephen L; Leangsuksun, Chokchai; He, X.

    2007-01-01

    This paper provides an overview of existing programming models for service-level high availability and investigates their differences, similarities, advantages, and disadvantages. Its goal is to help to improve reuse of code and to allow adaptation to quality of service requirements by using a uniform programming model description. It further aims at encouraging a discussion about these programming models and their provided quality of service, such as availability, performance, serviceability, usability, and applicability. Within this context, the presented research focuses on providing high availability for services running on head and service nodes of high-performance computing systems.

  20. High-Level Waste Vitrification Facility Feasibility Study

    SciTech Connect

    D. A. Lopez

    1999-08-01

    A ''Settlement Agreement'' between the Department of Energy and the State of Idaho mandates that all radioactive high-level waste now stored at the Idaho Nuclear Technology and Engineering Center will be treated so that it is ready to be moved out of Idaho for disposal by a compliance date of 2035. This report investigates vitrification treatment of the high-level waste in a High-Level Waste Vitrification Facility based on the assumption that no more New Waste Calcining Facility campaigns will be conducted after June 2000. Under this option, the sodium-bearing waste remaining in the Idaho Nuclear Technology and Engineering Center Tank Farm, and newly generated liquid waste produced between now and the start of 2013, will be processed using a different option, such as a Cesium Ion Exchange Facility. The cesium-saturated waste from this other option will be sent to the Calcine Solids Storage Facilities to be mixed with existing calcine. The calcine and cesium-saturated waste will be processed in the High-Level Waste Vitrification Facility by the end of calendar year 2035. In addition, the High-Level Waste Vitrification Facility will process all newly-generated liquid waste produced between 2013 and the end of 2035. Vitrification of this waste is an acceptable treatment method for complying with the Settlement Agreement. This method involves vitrifying the waste and pouring it into stainless-steel canisters that will be ready for shipment out of Idaho to a disposal facility by 2035. These canisters will be stored at the Idaho National Engineering and Environmental Laboratory until they are sent to a national geologic repository. The operating period for vitrification treatment will be from the end of 2015 through 2035.

  1. Long-term high-level waste technology

    NASA Astrophysics Data System (ADS)

    Cornman, W. R.

    1980-07-01

    This series of reports summarizes research and development studies on the immobilization of high level wastes from the chemical reprocessing of nuclear reactor fuels. Immobilization of the wastes (defense and commercial) consists of placing them in a high integrity form with a very low potential for radionuclide release. Immobilization of commercial wastes is being considered on a contingency basis in the event that reprocessing is resumed. The basic plan for meeting the goal of immobilization of the DOE high level wastes is: (1) to develop technology to support a realistic choice of waste form alternatives for each of the three DOE sites; (2) to develop product and processing technology with sufficient scaleup to provide design data for full scale facilities; and (3) to construct and operate the facilities.

  2. Extremely high urine arsenic level after remote seafood ingestion.

    PubMed

    Nañagas, Kristine A; Tormoehlen, Laura M

    2014-01-01

    Urine testing for heavy metal concentrations is increasingly performed in the outpatient setting as a part of laboratory evaluation for neuropathy. Abnormal urine arsenic levels due to dietary intake of organic arsenic can lead to unnecessary chelation therapy. A 54-year-old man underwent a 24-hour urine collection for heavy metal concentrations in evaluation of paresthesia of the right foot. The total arsenic level was 8880 μg/d with concentrations of 4749 μg/L and 3769 μg/g creatinine. He was urgently referred to the toxicology clinic for consideration of chelation therapy. History revealed consumption of 2 lobster tails 5 days before the testing. Speciation was then performed on the original urine specimen and revealed an organic arsenic concentration of 4332 μg/L. No inorganic or methylated arsenic was detected. Repeat testing after abstaining from seafood demonstrated a total arsenic level of 50 μg/d with concentrations of 30 μg/L and 21 μg/g creatinine. Our patient demonstrates the highest level of arsenobetaine reported in the literature, and this level is higher than expected for a person who had not consumed seafood for 5 days before testing. The high levels may be due to consumption of food that he did not recognize as containing arsenobetaine or that his clearance of arsenobetaine from the ingested lobster is slower than published ranges. This case demonstrates the importance of speciation when measuring urine arsenic levels to avoid unnecessary chelation therapy.

  3. High leptin levels are associated with migraine with aura.

    PubMed

    Pisanu, Claudia; Preisig, Martin; Castelao, Enrique; Glaus, Jennifer; Cunningham, Janet L; Del Zompo, Maria; Merikangas, Kathleen R; Schiöth, Helgi B; Mwinyi, Jessica

    2017-04-01

    Background Migraine is a prevalent disorder characterised by recurrent headache attacks preceded or accompanied by aura in a subgroup of patients. Migraine often occurs together with major depressive disorder (MDD). Alterations of adipokine levels have been reported both in migraine and in MDD. In this cross-sectional study, we aimed to assess the associations between serum leptin and adiponectin levels and migraine or migraine subtypes. Analyses were adjusted for a lifetime history of MDD in order to investigate the association between adipokines and migraine under consideration of depression status. Methods We included 3025 participants from the CoLaus/PsyCoLaus study. The impact of leptin and adiponectin levels on a diagnosis of migraine was analysed by binary regression analyses, adjusting for variables known to influence adipokine levels. Subgroup analyses were conducted based on the presence of aura. Results Crude leptin levels were significantly higher in subjects with migraine than controls (Mann-Whitney U = 515,102, p = 6 × 10(-7)). When performing adjusted analyses, leptin levels were found to be significantly higher in subjects with migraine (odds ratio = 1.22, p = 0.024) and migraine with aura (odds ratio = 1.34, p = 0.004). Conclusion High leptin levels might play a role in the pathogenesis of migraine and migraine with aura.

  4. [Medical image retrieval by high level semantic features and low level content features of image].

    PubMed

    Xie, Tianwen; Tang, Weijun; Zhao, Qiufeng; Zhao, Jiaao

    2009-12-01

    Content-based image retrieval aims at searching the similar images using low level features,and medical image retrieval needs it for the retrieval of similar images. Medical images contain not only a lot of content data, but also a lot of semantic information. This paper presents an approach by combining digital imaging and communications in medicine (DICOM) features and low level features to perform retrieval on medical image databases. At the first step, the semantic information is extracted from DICOM header for the pre-filtering of the images, and then dual-tree complex wavelet transfrom(DT-CWT) features of pre-filtered images and example images are extracted to retrieve similar images. Experimental results show that by combining the high level semantics (DICOM features) and low level content features (texture) the retrieval time is reduced and the performance of medical image retrieval is increased.

  5. Very-high-level neutral-beam control system

    SciTech Connect

    Elischer, V.; Jacobson, V.; Theil, E.

    1981-10-01

    As increasing numbers of neutral beams are added to fusion machines, their operation can consume a significant fraction of a facility's total resources. LBL has developed a very high level control system that allows a neutral beam injector to be treated as a black box with just 2 controls: one to set the beam power and one to set the pulse duration. This 2 knob view allows simple operation and provides a natural base for implementing even higher level controls such as automatic source conditioning.

  6. High plasma adenosine levels in overweight/obese pregnant women.

    PubMed

    Badillo, Priscila; Salgado, Paola; Bravo, Patricia; Guevara, Katherine; Acurio, Jesenia; Gonzalez, Maria Angelica; Oyarzun, Carlos; San Martin, Rody; Escudero, Carlos

    2017-07-18

    We aim to investigate whether overweight/obese pregnant women have elevated plasma levels of adenosine associated with increased consumption of high-calorie food. Sixty women were included. They were divided into lean (n = 23 and n = 12) or overweight/obese (n = 7 and n = 18) non-pregnant and pregnant women, respectively. Clinical records and maternal blood samples were collected after informed consent. A self-reported dietary questionnaire was also completed. Plasma adenosine levels were determined with high-performance liquid chromatography. Biochemical parameters, including glucose, total protein, and lipid profile, were determined using standard colorimetric assays. Adenosine levels were higher in pregnant women than in non-pregnant women (18.7 ± 1.6 vs 10.8 ± 1.3 nM/μg protein, respectively, p < 0.0001). Overweight/obese pregnant women (21.9 ± 2.5 nM/μg protein) exhibited higher adenosine levels than lean pregnant (14.5 ± 1.0 nM/μg protein, p = 0.04) or non-pregnant women (11.7 ± 1.5 nM/μg protein, p = 0.0005). Also, pregnant women with elevated weight gain exhibited higher (26.2 ± 3.7 nM/μg protein) adenosine levels than those with adequate weight gain (14.9 ± 1.4 nM/μg protein, p = 0.03). These differences were not statistically significant compared with those of pregnant women with reduced weight gain (17.4 ± 2.1 nM/μg protein, p = 0.053). Body mass index and adenosine only in pregnant women were positively correlated (r = 0.39, p = 0.02). While, polyunsaturated fatty acid (PUFA) consumption was negatively correlated with plasma adenosine levels only in non-pregnant women (r = -0.33, p = 0.03). Pregnancy is associated with high plasma adenosine levels, which are further elevated in pregnant women who are overweight/obese. High PUFA intake might reduce plasma adenosine levels in non-pregnant women.

  7. High Prepubertal Leptin Levels Are Associated With Earlier Menarcheal Age.

    PubMed

    Gavela-Pérez, Teresa; Navarro, Pilar; Soriano-Guillén, Leandro; Garcés, Carmen

    2016-08-01

    Given the interplay between metabolic status and the reproductive system, factors governing energy homeostasis could influence the timing of pubertal onset. The aim of this longitudinal study was to determine the influence of metabolic peptides associated to adiposity on menarcheal age. The sample population included 168 girls. Anthropometric and biochemical variables were measured at a prepubertal baseline (6-8 years old) and 7 years later. Leptin and adiponectin levels were determined by Enzyme-Linked ImmunoSorbent Assay, insulin by radioimmunoassay and ghrelin by a multiplexed bead immunoassay. Adjusted for body mass index, Spearman correlation analysis revealed a significant negative association between prepubertal leptin levels and menarche. The highest prepubertal leptin levels were observed in overweight girls with the earliest menarche. Menarcheal age was significantly associated with variation of adiponectin levels between both baseline and follow-up. Multiple regression analysis showed that prepubertal leptin was the only significant contributing factor, predicting 15.3% of variation in menarcheal age. High leptin levels in prepubertal girls are associated with earlier menarcheal age, independent of body mass index. However, the relationship of ghrelin, insulin, or adiponectin levels with menarcheal age is not evident in our population. Copyright © 2016 Society for Adolescent Health and Medicine. Published by Elsevier Inc. All rights reserved.

  8. Mammut: High-level management of system knobs and sensors

    NASA Astrophysics Data System (ADS)

    De Sensi, Daniele; Torquati, Massimo; Danelutto, Marco

    Managing low-level architectural features for controlling performance and power consumption is a growing demand in the parallel computing community. Such features include, but are not limited to: energy profiling, platform topology analysis, CPU cores disabling and frequency scaling. However, these low-level mechanisms are usually managed by specific tools, without any interaction between each other, thus hampering their usability. More important, most existing tools can only be used through a command line interface and they do not provide any API. Moreover, in most cases, they only allow monitoring and managing the same machine on which the tools are used. MAMMUT provides and integrates architectural management utilities through a high-level and easy-to-use object-oriented interface. By using MAMMUT, is possible to link together different collected information and to exploit them on both local and remote systems, to build architecture-aware applications.

  9. Landau-Level Splitting in Graphene in High Magnetic Fields

    NASA Astrophysics Data System (ADS)

    Zhang, Y.; Jiang, Z.; Small, J. P.; Purewal, M. S.; Tan, Y.-W.; Fazlollahi, M.; Chudow, J. D.; Jaszczak, J. A.; Stormer, H. L.; Kim, P.

    2006-04-01

    The quantum Hall (QH) effect in two-dimensional electrons and holes in high quality graphene samples is studied in strong magnetic fields up to 45 T. QH plateaus at filling factors ν=0,±1,±4 are discovered at magnetic fields B>20T, indicating the lifting of the fourfold degeneracy of the previously observed QH states at ν=±4(|n|+1/2), where n is the Landau-level index. In particular, the presence of the ν=0,±1 QH plateaus indicates that the Landau level at the charge neutral Dirac point splits into four sublevels, lifting sublattice and spin degeneracy. The QH effect at ν=±4 is investigated in a tilted magnetic field and can be attributed to lifting of the spin degeneracy of the n=1 Landau level.

  10. FLUIDIZED BED STEAM REFORMING ENABLING ORGANIC HIGH LEVEL WASTE DISPOSAL

    SciTech Connect

    Williams, M

    2008-05-09

    Waste streams planned for generation by the Global Nuclear Energy Partnership (GNEP) and existing radioactive High Level Waste (HLW) streams containing organic compounds such as the Tank 48H waste stream at Savannah River Site have completed simulant and radioactive testing, respectfully, by Savannah River National Laboratory (SRNL). GNEP waste streams will include up to 53 wt% organic compounds and nitrates up to 56 wt%. Decomposition of high nitrate streams requires reducing conditions, e.g. provided by organic additives such as sugar or coal, to reduce NOX in the off-gas to N2 to meet Clean Air Act (CAA) standards during processing. Thus, organics will be present during the waste form stabilization process regardless of the GNEP processes utilized and exists in some of the high level radioactive waste tanks at Savannah River Site and Hanford Tank Farms, e.g. organics in the feed or organics used for nitrate destruction. Waste streams containing high organic concentrations cannot be stabilized with the existing HLW Best Developed Available Technology (BDAT) which is HLW vitrification (HLVIT) unless the organics are removed by pretreatment. The alternative waste stabilization pretreatment process of Fluidized Bed Steam Reforming (FBSR) operates at moderate temperatures (650-750 C) compared to vitrification (1150-1300 C). The FBSR process has been demonstrated on GNEP simulated waste and radioactive waste containing high organics from Tank 48H to convert organics to CAA compliant gases, create no secondary liquid waste streams and create a stable mineral waste form.

  11. Ionization chamber for measurements of high-level tritium gas

    SciTech Connect

    Carstens, D.H.W.; David, W.R.

    1980-01-01

    The construction and calibration of a simple ionization-chamber apparatus for measurement of high level tritium gas is described. The apparatus uses an easily constructed but rugged chamber containing the unknown gas and an inexpensive digital multimeter for measuring the ion current. The equipment after calibration is suitable for measuring 0.01 to 100% tritium gas in hydrogen-helium mixes with an accuracy of a few percent. At both the high and low limits of measurements deviations from the predicted theoretical current are observed. These are briefly discussed.

  12. RETENTION OF SULFATE IN HIGH LEVEL RADIOACTIVE WASTE GLASS

    SciTech Connect

    Fox, K.

    2010-09-07

    High level radioactive wastes are being vitrified at the Savannah River Site for long term disposal. Many of the wastes contain sulfate at concentrations that can be difficult to retain in borosilicate glass. This study involves efforts to optimize the composition of a glass frit for combination with the waste to improve sulfate retention while meeting other process and product performance constraints. The fabrication and characterization of several series of simulated waste glasses are described. The experiments are detailed chronologically, to provide insight into part of the engineering studies used in developing frit compositions for an operating high level waste vitrification facility. The results lead to the recommendation of a specific frit composition and a concentration limit for sulfate in the glass for the next batch of sludge to be processed at Savannah River.

  13. Nondestructive examination of DOE high-level waste storage tanks

    SciTech Connect

    Bush, S.; Bandyopadhyay, K.; Kassir, M.; Mather, B.; Shewmon, P.; Streicher, M.; Thompson, B.; van Rooyen, D.; Weeks, J.

    1995-05-01

    A number of DOE sites have buried tanks containing high-level waste. Tanks of particular interest am double-shell inside concrete cylinders. A program has been developed for the inservice inspection of the primary tank containing high-level waste (HLW), for testing of transfer lines and for the inspection of the concrete containment where possible. Emphasis is placed on the ultrasonic examination of selected areas of the primary tank, coupled with a leak-detection system capable of detecting small leaks through the wall of the primary tank. The NDE program is modelled after ASME Section XI in many respects, particularly with respects to the sampling protocol. Selected testing of concrete is planned to determine if there has been any significant degradation. The most probable failure mechanisms are corrosion-related so that the examination program gives major emphasis to possible locations for corrosion attack.

  14. Review of High Level Waste Tanks Ultrasonic Inspection Data

    SciTech Connect

    Wiersma, B

    2006-03-09

    A review of the data collected during ultrasonic inspection of the Type I high level waste tanks has been completed. The data was analyzed for relevance to the possibility of vapor space corrosion and liquid/air interface corrosion. The review of the Type I tank UT inspection data has confirmed that the vapor space general corrosion is not an unusually aggressive phenomena and correlates well with predicted corrosion rates for steel exposed to bulk solution. The corrosion rates are seen to decrease with time as expected. The review of the temperature data did not reveal any obvious correlations between high temperatures and the occurrences of leaks. The complex nature of temperature-humidity interaction, particularly with respect to vapor corrosion requires further understanding to infer any correlation. The review of the waste level data also did not reveal any obvious correlations.

  15. Life Extension of Aging High-Level Waste Tanks

    SciTech Connect

    Bryson, D.; Callahan, V.; Ostrom, M.; Bryan, W.; Berman, H.

    2002-02-26

    The Double Shell Tanks (DSTs) play a critical role in the Hanford High-Level Waste Treatment Complex, and therefore activities are underway to protect and better understand these tanks. The DST Life Extension Program is focused on both tank life extension and on evaluation of tank integrity. Tank life extension activities focus on understanding tank failure modes and have produced key chemistry and operations controls to minimize tank corrosion and extend useful tank life. Tank integrity program activities have developed and applied key technologies to evaluate the condition of the tank structure and predict useful tank life. Program results to date indicate that DST useful life can be extended well beyond the original design life and allow the existing tanks to fill a critical function within the Hanford High-Level Waste Treatment Complex. In addition the tank life may now be more reliably predicted, facilitating improved planning for the use and possible future replacement of these tanks.

  16. Evaluation and selection of candidate high-level waste forms

    SciTech Connect

    Bernadzikowski, T. A.; Allender, J. S.; Butler, J. L.; Gordon, D. E.; Gould, Jr., T. H.; Stone, J. A.

    1982-03-01

    Seven candidate waste forms being developed under the direction of the Department of Energy's National High-Level Waste (HLW) Technology Program, were evaluated as potential media for the immobilization and geologic disposal of high-level nuclear wastes. The evaluation combined preliminary waste form evaluations conducted at DOE defense waste-sites and independent laboratories, peer review assessments, a product performance evaluation, and a processability analysis. Based on the combined results of these four inputs, two of the seven forms, borosilicate glass and a titanate based ceramic, SYNROC, were selected as the reference and alternative forms for continued development and evaluation in the National HLW Program. Both the glass and ceramic forms are viable candidates for use at each of the DOE defense waste-sites; they are also potential candidates for immobilization of commercial reprocessing wastes. This report describes the waste form screening process, and discusses each of the four major inputs considered in the selection of the two forms.

  17. Long-term high-level waste technology

    NASA Astrophysics Data System (ADS)

    Corman, W. R.

    1981-08-01

    Work performed at sites to immobilize high-level radioactive wastes is described. Program management and support with subtasks of management and budget, environmental and safety assessments, waste preparation, storage or disposal; waste retrieval, separation and concentration are discussed. Waste fixation and characterization, process and equipment development, final handling, canister development and characterization and onsite storage or disposal are also reported. Event trees defining possible accidents were completed in a safety assessment of continued in-tank storage of high-level waste. Two low-cost waste forms (tailored concrete and bitumen) were investigated as candidate immobilization forms. Comparative impact tests and leaching tests were also conducted on glasses, ceramics, and concretes. A process design description was written for the tailored ceramic process.

  18. Multipurpose optimization models for high level waste vitrification

    SciTech Connect

    Hoza, M.

    1994-08-01

    Optimal Waste Loading (OWL) models have been developed as multipurpose tools for high-level waste studies for the Tank Waste Remediation Program at Hanford. Using nonlinear programming techniques, these models maximize the waste loading of the vitrified waste and optimize the glass formers composition such that the glass produced has the appropriate properties within the melter, and the resultant vitrified waste form meets the requirements for disposal. The OWL model can be used for a single waste stream or for blended streams. The models can determine optimal continuous blends or optimal discrete blends of a number of different wastes. The OWL models have been used to identify the most restrictive constraints, to evaluate prospective waste pretreatment methods, to formulate and evaluate blending strategies, and to determine the impacts of variability in the wastes. The OWL models will be used to aid in the design of frits and the maximize the waste in the glass for High-Level Waste (HLW) vitrification.

  19. FPGA based compute nodes for high level triggering in PANDA

    NASA Astrophysics Data System (ADS)

    Kühn, W.; Gilardi, C.; Kirschner, D.; Lang, J.; Lange, S.; Liu, M.; Perez, T.; Yang, S.; Schmitt, L.; Jin, D.; Li, L.; Liu, Z.; Lu, Y.; Wang, Q.; Wei, S.; Xu, H.; Zhao, D.; Korcyl, K.; Otwinowski, J. T.; Salabura, P.; Konorov, I.; Mann, A.

    2008-07-01

    PANDA is a new universal detector for antiproton physics at the HESR facility at FAIR/GSI. The PANDA data acquisition system has to handle interaction rates of the order of 107/s and data rates of several 100 Gb/s. FPGA based compute nodes with multi-Gb/s bandwidth capability using the ATCA architecture are designed to handle tasks such as event building, feature extraction and high level trigger processing. Data connectivity is provided via optical links as well as multiple Gb Ethernet ports. The boards will support trigger algorithms such us pattern recognition for RICH detectors, EM shower analysis, fast tracking algorithms and global event characterization. Besides VHDL, high level C-like hardware description languages will be considered to implement the firmware.

  20. Management of data quality of high level waste characterization

    SciTech Connect

    Winters, W.I., Westinghouse Hanford

    1996-06-12

    Over the past 10 years, the Hanford Site has been transitioning from nuclear materials production to Site cleanup operations. High-level waste characterization at the Hanford Site provides data to support present waste processing operations, tank safety programs, and future waste disposal programs. Quality elements in the high-level waste characterization program will be presented by following a sample through the data quality objective, sampling, laboratory analysis and data review process. Transition from production to cleanup has resulted in changes in quality systems and program; the changes, as well as other issues in these quality programs, will be described. Laboratory assessment through quality control and performance evaluation programs will be described, and data assessments in the laboratory and final reporting in the tank characterization reports will be discussed.

  1. Life Extension of Aging High Level Waste (HLW) Tanks

    SciTech Connect

    BRYSON, D.

    2002-02-04

    The Double Shell Tanks (DSTs) play a critical role in the Hanford High-Level Waste Treatment Complex, and therefore activities are underway to protect and better understand these tanks. The DST Life Extension Program is focused on both tank life extension and on evaluation of tank integrity. Tank life extension activities focus on understanding tank failure modes and have produced key chemistry and operations controls to minimize tank corrosion and extend useful tank life. Tank integrity program activities have developed and applied key technologies to evaluate the condition of the tank structure and predict useful tank life. Program results to date indicate that DST useful life can be extended well beyond the original design life and allow the existing tanks to fill a critical function within the Hanford High-Level Waste Treatment Complex. In addition the tank life may now be more reliably predicted, facilitating improved planning for the use and possible future replacement of these tanks.

  2. High-level waste management technology program plan

    SciTech Connect

    Harmon, H.D.

    1995-01-01

    The purpose of this plan is to document the integrated technology program plan for the Savannah River Site (SRS) High-Level Waste (HLW) Management System. The mission of the SRS HLW System is to receive and store SRS high-level wastes in a see and environmentally sound, and to convert these wastes into forms suitable for final disposal. These final disposal forms are borosilicate glass to be sent to the Federal Repository, Saltstone grout to be disposed of on site, and treated waste water to be released to the environment via a permitted outfall. Thus, the technology development activities described herein are those activities required to enable successful accomplishment of this mission. The technology program is based on specific needs of the SRS HLW System and organized following the systems engineering level 3 functions. Technology needs for each level 3 function are listed as reference, enhancements, and alternatives. Finally, FY-95 funding, deliverables, and schedules are s in Chapter IV with details on the specific tasks that are funded in FY-95 provided in Appendix A. The information in this report represents the vision of activities as defined at the beginning of the fiscal year. Depending on emergent issues, funding changes, and other factors, programs and milestones may be adjusted during the fiscal year. The FY-95 SRS HLW technology program strongly emphasizes startup support for the Defense Waste Processing Facility and In-Tank Precipitation. Closure of technical issues associated with these operations has been given highest priority. Consequently, efforts on longer term enhancements and alternatives are receiving minimal funding. However, High-Level Waste Management is committed to participation in the national Radioactive Waste Tank Remediation Technology Focus Area. 4 refs., 5 figs., 9 tabs.

  3. A high resolution water level forecast for the German Bight

    NASA Astrophysics Data System (ADS)

    Niehüser, Sebastian; Dangendorf, Sönke; Arns, Arne; Jensen, Jürgen

    2016-04-01

    Many coastal regions worldwide are potentially endangered by storm surges which can cause disastrous damages and loss of life. Due to climate change induced sea level rise, an accumulation of such events is expected by the end of the 21th century. Therefore, advanced storm surge warnings are needed to be prepared when another storm surge hits the coast. In the shallow southeastern North Sea these storm surge warnings are nowadays routinely provided for selected tide gauge locations along a coastline through state-of-the-art forecast systems, which are based on a coupled system of empirical tidal predictions and numerical storm surge forecasts. Along the German North Sea coastline, the Federal Maritime and Hydrographic Agency in cooperation with the German Weather Service is responsible for the storm surge warnings. They provide accurate, high frequency and real-time water level forecasts for up to six days ahead at selected tide gauge sites via internet, telephone and broadcast. Since water levels along the German North Sea coastline are dominated by shallow water effects and a very complex bathymetric structure of the seabed, the pointwise forecast is not necessarily transferable to un-gauged areas between the tide gauges. Here we aim to close this existing gap and develop water level forecasts with a high spatial (continuously with a resolution of at least 1 kilometer) as well as a high temporal (at least 15-minute values) resolution along the entire German North Sea coastline. We introduce a new methodology for water level forecasts which combines empirical or statistical and numerical models. While the tidal forecast is performed by non-parametric interpolation techniques between un-gauged and gauged sites, storm surges are estimated on the basis of statistical/empirical storm surge formulas taken from a numerical model hindcast. The procedure will be implemented in the operational mode forced with numerical weather forecasts.

  4. High Level Information Fusion (HLIF) with nested fusion loops

    NASA Astrophysics Data System (ADS)

    Woodley, Robert; Gosnell, Michael; Fischer, Amber

    2013-05-01

    Situation modeling and threat prediction require higher levels of data fusion in order to provide actionable information. Beyond the sensor data and sources the analyst has access to, the use of out-sourced and re-sourced data is becoming common. Through the years, some common frameworks have emerged for dealing with information fusion—perhaps the most ubiquitous being the JDL Data Fusion Group and their initial 4-level data fusion model. Since these initial developments, numerous models of information fusion have emerged, hoping to better capture the human-centric process of data analyses within a machine-centric framework. 21st Century Systems, Inc. has developed Fusion with Uncertainty Reasoning using Nested Assessment Characterizer Elements (FURNACE) to address challenges of high level information fusion and handle bias, ambiguity, and uncertainty (BAU) for Situation Modeling, Threat Modeling, and Threat Prediction. It combines JDL fusion levels with nested fusion loops and state-of-the-art data reasoning. Initial research has shown that FURNACE is able to reduce BAU and improve the fusion process by allowing high level information fusion (HLIF) to affect lower levels without the double counting of information or other biasing issues. The initial FURNACE project was focused on the underlying algorithms to produce a fusion system able to handle BAU and repurposed data in a cohesive manner. FURNACE supports analyst's efforts to develop situation models, threat models, and threat predictions to increase situational awareness of the battlespace. FURNACE will not only revolutionize the military intelligence realm, but also benefit the larger homeland defense, law enforcement, and business intelligence markets.

  5. Mixing Processes in High-Level Waste Tanks - Final Report

    SciTech Connect

    Peterson, P.F.

    1999-05-24

    The mixing processes in large, complex enclosures using one-dimensional differential equations, with transport in free and wall jets is modeled using standard integral techniques. With this goal in mind, we have constructed a simple, computationally efficient numerical tool, the Berkeley Mechanistic Mixing Model, which can be used to predict the transient evolution of fuel and oxygen concentrations in DOE high-level waste tanks following loss of ventilation, and validate the model against a series of experiments.

  6. Case for retrievable high-level nuclear waste disposal

    USGS Publications Warehouse

    Roseboom, Eugene H.

    1994-01-01

    Plans for the nation's first high-level nuclear waste repository have called for permanently closing and sealing the repository soon after it is filled. However, the hydrologic environment of the proposed site at Yucca Mountain, Nevada, should allow the repository to be kept open and the waste retrievable indefinitely. This would allow direct monitoring of the repository and maintain the options for future generations to improve upon the disposal methods or use the uranium in the spent fuel as an energy resource.

  7. Automatic rule generation for high-level vision

    NASA Technical Reports Server (NTRS)

    Rhee, Frank Chung-Hoon; Krishnapuram, Raghu

    1992-01-01

    Many high-level vision systems use rule-based approaches to solving problems such as autonomous navigation and image understanding. The rules are usually elaborated by experts. However, this procedure may be rather tedious. In this paper, we propose a method to generate such rules automatically from training data. The proposed method is also capable of filtering out irrelevant features and criteria from the rules.

  8. High-Speed, Low-Level Flight: Aircrew Factors

    DTIC Science & Technology

    1980-03-01

    clear from the material presented that the major problems currently encountered in high-speed, low-level flight are ’ride-bumpiness’, excessive workload...which we have for centuries, colonized, administered, developed, explored and loved. The unexpected drying up of preferential sources of raw materials and...definition means assuming responsibility for providing them with the material means necessary to carry the assigned mission, one can nevertheless say that the

  9. Learning high-level features for chord recognition using Autoencoder

    NASA Astrophysics Data System (ADS)

    Phongthongloa, Vilailukkana; Kamonsantiroj, Suwatchai; Pipanmaekaporn, Luepol

    2016-07-01

    Chord transcription is valuable to do by itself. It is known that the manual transcription of chords is very tiresome, time-consuming. It requires, moreover, musical knowledge. Automatic chord recognition has recently attracted a number of researches in the Music Information Retrieval field. It has known that a pitch class profile (PCP) is the commonly signal representation of musical harmonic analysis. However, the PCP may contain additional non-harmonic noise such as harmonic overtones and transient noise. The problem of non-harmonic might be generating the sound energy in term of frequency more than the actual notes of the respective chord. Autoencoder neural network may be trained to learn a mapping from low level feature to one or more higher-level representation. These high-level representations can explain dependencies of the inputs and reduce the effect of non-harmonic noise. Then these improve features are fed into neural network classifier. The proposed high-level musical features show 80.90% of accuracy. The experimental results have shown that the proposed approach can achieve better performance in comparison with other based method.

  10. Local acceptance of a high-level nuclear waste repository.

    PubMed

    Sjöberg, Lennart

    2004-06-01

    The siting of nuclear waste facilities has been very difficult in all countries. Recent experience in Sweden indicates, however, that it may be possible, under certain circumstances, to gain local support for the siting of a high-level nuclear waste (HLNW) repository. The article reports on a study of attitudes and risk perceptions of people living in four municipalities in Sweden where HLNW siting was being intensely discussed at the political level, in media, and among the public. Data showed a relatively high level of consensus on acceptability of at least further investigation of the issue; in two cases local councils have since voted in favor of a go-ahead, and in one case only a very small majority defeated the issue. Models of policy attitudes showed that these were related to attitude to nuclear power, attributes of the perceived HLNW risk, and trust. Factors responsible for acceptance are discussed at several levels. One is the attitude to nuclear power, which is becoming more positive, probably because no viable alternatives are in sight. Other factors have to do with the extensive information programs conducted in these municipalities, and with the logical nature of the conclusion that they would be good candidates for hosting the national HLNW repository.

  11. Handbook of high-level radioactive waste transportation

    SciTech Connect

    Sattler, L.R.

    1992-10-01

    The High-Level Radioactive Waste Transportation Handbook serves as a reference to which state officials and members of the general public may turn for information on radioactive waste transportation and on the federal government`s system for transporting this waste under the Civilian Radioactive Waste Management Program. The Handbook condenses and updates information contained in the Midwestern High-Level Radioactive Waste Transportation Primer. It is intended primarily to assist legislators who, in the future, may be called upon to enact legislation pertaining to the transportation of radioactive waste through their jurisdictions. The Handbook is divided into two sections. The first section places the federal government`s program for transporting radioactive waste in context. It provides background information on nuclear waste production in the United States and traces the emergence of federal policy for disposing of radioactive waste. The second section covers the history of radioactive waste transportation; summarizes major pieces of legislation pertaining to the transportation of radioactive waste; and provides an overview of the radioactive waste transportation program developed by the US Department of Energy (DOE). To supplement this information, a summary of pertinent federal and state legislation and a glossary of terms are included as appendices, as is a list of publications produced by the Midwestern Office of The Council of State Governments (CSG-MW) as part of the Midwestern High-Level Radioactive Waste Transportation Project.

  12. Materials Science of High-Level Nuclear Waste Immobilization

    SciTech Connect

    Weber, William J.; Navrotsky, Alexandra; Stefanovsky, S. V.; Vance, E. R.; Vernaz, Etienne Y.

    2009-01-09

    With the increasing demand for the development of more nuclear power comes the responsibility to address the technical challenges of immobilizing high-level nuclear wastes in stable solid forms for interim storage or disposition in geologic repositories. The immobilization of high-level nuclear wastes has been an active area of research and development for over 50 years. Borosilicate glasses and complex ceramic composites have been developed to meet many technical challenges and current needs, although regulatory issues, which vary widely from country to country, have yet to be resolved. Cooperative international programs to develop advanced proliferation-resistant nuclear technologies to close the nuclear fuel cycle and increase the efficiency of nuclear energy production might create new separation waste streams that could demand new concepts and materials for nuclear waste immobilization. This article reviews the current state-of-the-art understanding regarding the materials science of glasses and ceramics for the immobilization of high-level nuclear waste and excess nuclear materials and discusses approaches to address new waste streams.

  13. Bacteriologic testing of endoscopes after high-level disinfection.

    PubMed

    Rejchrt, Stanislav; Cermák, Pavel; Pavlatová, Ludmila; McKová, Eva; Bures, Jan

    2004-07-01

    There are no definitive data available concerning microbiologic safety of prolonged endoscope storage after reprocessing and disinfection. This study evaluated the durability of high-level disinfection of endoscopes stored in a dust-proof cabinet for 5 days. Three different types of endoscopes (upper endoscopes, duodenoscopes, colonoscopes) were tested. After completion of the endoscopic procedure, endoscopes were subjected to an initial decontamination, followed by manual cleaning with the endoscope immersed in detergent. The endoscopes then were placed in an automatic reprocessor that provides high-level disinfection. They then were stored by hanging in a dust-proof cabinet. Bacteriologic samples were obtained from the surface of the endoscopes, the openings for the piston valves, and the accessory channel daily for 5 days, and by flush-through (combined with brushing) from the accessory channels after 5 days of storage. Samples were cultured for all types of aerobic and anaerobic bacteria, including bacterial spores, and for Candida species. For all assays, all endoscopes were bacteria-free immediately after high-level disinfection. Only 4 assays (of 135) were positive during the subsequent 5-day assessment (skin bacteria cultured from endoscope surfaces). All flush-through samples were sterile. When endoscope reprocessing guidelines are strictly observed and endoscopes are stored in appropriate cabinets for up to 5 days, reprocessing before use may not be necessary.

  14. High Levels of Molecular Chlorine found in the Arctic Atmosphere

    NASA Astrophysics Data System (ADS)

    Liao, J.; Huey, L. G.; Liu, Z.; Tanner, D.; Cantrell, C. A.; Orlando, J. J.; Flocke, F. M.; Shepson, P. B.; Weinheimer, A. J.; Hall, S. R.; Beine, H.; Wang, Y.; Ingall, E. D.; Thompson, C. R.; Hornbrook, R. S.; Apel, E. C.; Fried, A.; Mauldin, L.; Smith, J. N.; Staebler, R. M.; Neuman, J. A.; Nowak, J. B.

    2014-12-01

    Chlorine radicals are a strong atmospheric oxidant, particularly in polar regions where levels of hydroxyl radicals can be quite low. In the atmosphere, chlorine radicals expedite the degradation of methane and tropospheric ozone and the oxidation of mercury to more toxic forms. Here, we present direct measurements of molecular chlorine levels in the Arctic marine boundary layer in Barrow, Alaska, collected in the spring of 2009 over a six-week period using chemical ionization mass spectrometry. We detected high levels of molecular chlorine of up to 400 pptv. Concentrations peaked in the early morning and late afternoon and fell to near-zero levels at night. Average daytime molecular chlorine levels were correlated with ozone concentrations, suggesting that sunlight and ozone are required for molecular chlorine formation. Using a time-dependent box model, we estimated that the chlorine radicals produced from the photolysis of molecular chlorine on average oxidized more methane than hydroxyl radicals and enhanced the abundance of short-lived peroxy radicals. Elevated hydroperoxyl radical levels, in turn, promoted the formation of hypobromous acid, which catalyzed mercury oxidation and the breakdown of tropospheric ozone. Therefore, we propose that molecular chlorine exerts a significant effect on the atmospheric chemistry in the Arctic. While the formation mechanisms of molecular chlorine are not yet understood, the main potential sources of chlorine include snowpack, sea salt, and sea ice. There is recent evidence of molecular halogen (Br2 and Cl2) formation in the Arctic snowpack. The coverage and composition of the snow may control halogen chemistry in the Arctic. Changes of sea ice and snow cover in the changing climate may affect air-snow-ice interaction and have a significant impact on the levels of radicals, ozone, mercury and methane in the Arctic troposphere.

  15. Multiply spliced env and nef transcripts of simian immunodeficiency virus from West African green monkey (SIVagm-sab).

    PubMed

    Bibollet-Ruche, F; Cuny, G; Pourrut, X; Brengues, C; Galat-Luong, A; Galat, G; Delaporte, E

    1998-04-10

    We have characterized the spliced transcripts of nef and envelope genes of SIVagm from African green monkey of the sabaeus subspecies. Most of the transcripts we have studied, representing the most abundant mRNA species in our assay, have undergone a specific splicing event that removes a part of the trans-activation response (TAR) element. This region is predicted to form a stable secondary structure (four stem-loop elements in SIVagm-sab) that affects the trans-activation of viral gene expression by Tat and the translation of the viral transcripts. Contrary to what is observed in other viruses, in which this R-region splicing has also been described (e.g., HIV-2), the LTR splicing in SIVagm-sab removes part of the first stem-loop and the following ones, nearly completely disrupting the TAR element secondary structure. Because LTR splicing seems to be a conserved feature among the strains we have characterized, these results suggest that this phenomenon could have important consequences for virus replication, pathogenicity, and latency.

  16. Identification of new and unusual rev and nef transcripts expressed by an HIV type 1 primary isolate.

    PubMed

    Vega, Yolanda; Delgado, Elena; Carrera, Cristina; Nebreda, Paloma; Fernández-García, Aurora; Cuevas, María Teresa; Pérez-Álvarez, Lucía; Thomson, Michael M

    2013-07-01

    We analyzed RNA splice site usage in three HIV-1 subtype B primary isolates through reverse transcriptase polymerase chain reaction (RT-PCR) amplification of spliced RNAs using a fluorescently labeled primer, with computerized size determination and quantification of PCR products, which were also identified by clone sequencing. In one isolate, P2149-3, unusual and unreported spliced transcripts were detected. This isolate preferentially used for rev RNA generation a 3' splice site (3'ss) located five nucleotides upstream of A4a, previously identified only in a T cell line-adapted virus and in a group O isolate, and designated A4d. P2149-3 also used an unreported 3'ss for rev RNA generation, designated A4h, located 20 nucleotides upstream of 3'ss A4c. Additionally, unusual nef RNAs using 3'ss A5a and A7a and with exon composition 1.3.7 were identified. The identification of several unusual and unreported spliced transcripts in an HIV-1 primary isolate suggests a greater diversity of splice site usage in HIV-1 than previously appreciated.

  17. Identification of New and Unusual rev and nef Transcripts Expressed by an HIV Type 1 Primary Isolate

    PubMed Central

    Vega, Yolanda; Delgado, Elena; Carrera, Cristina; Nebreda, Paloma; Fernández-García, Aurora; Cuevas, María Teresa; Pérez-Álvarez, Lucía

    2013-01-01

    Abstract We analyzed RNA splice site usage in three HIV-1 subtype B primary isolates through reverse transcriptase polymerase chain reaction (RT-PCR) amplification of spliced RNAs using a fluorescently labeled primer, with computerized size determination and quantification of PCR products, which were also identified by clone sequencing. In one isolate, P2149-3, unusual and unreported spliced transcripts were detected. This isolate preferentially used for rev RNA generation a 3′ splice site (3′ss) located five nucleotides upstream of A4a, previously identified only in a T cell line-adapted virus and in a group O isolate, and designated A4d. P2149-3 also used an unreported 3′ss for rev RNA generation, designated A4h, located 20 nucleotides upstream of 3′ss A4c. Additionally, unusual nef RNAs using 3′ss A5a and A7a and with exon composition 1.3.7 were identified. The identification of several unusual and unreported spliced transcripts in an HIV-1 primary isolate suggests a greater diversity of splice site usage in HIV-1 than previously appreciated. PMID:23540799

  18. In this issue: Nef, a lingering problem; are there better immunoglobulins for human use; and Takayasu arteritis, still a mystery.

    PubMed

    Bot, Adrian

    2012-12-01

    The current issue of the International Reviews of Immunology brings the latest in three different areas of basic and clinical immunology. First, there is the lingering question of residual HIV-related pathology in patients on chronic antiretroviral therapy. The specific role of Nef in the macrophage-mediated disease manifested through lymphoma, metabolic disease and neurological disorder, is extensively discussed. A second topic is a clinical immunology one, a critical perspective on the efficacy and safety profile of various preparations of immunoglobulin products currently prescribed for a range of immunodeficiencies. Surprisingly, the authors showed that while the available preparations are equivalent from efficacy standpoint, they differ from the standpoint of toxicity. The third subject is again, in the arena of clinical immunology and deals with a relatively rare yet extremely puzzling disease -Takayasu arteritis -with a pathogenesis that needs elucidation and a dire need for better treatments. The authors provide a state of the art in terms of genetic association, other factors possibly involved in the onset and progression of this unusual inflammatory disease of large arteries, and provide a perspective on current standard of care and potential usefulness of TNF-α blockade as therapy for Takayasu arteritis.

  19. Association between high cystatin C levels and carotid atherosclerosis

    PubMed Central

    Kobayashi, Toshiyuki; Yokokawa, Hirohide; Fujibayashi, Kazutoshi; Haniu, Tomomi; Hisaoka, Teruhiko; Fukuda, Hiroshi; Naito, Toshio

    2017-01-01

    AIM To investigate the association between carotid atherosclerosis and cystatin C (CysC) and to determine the optimal CysC cut-off value. METHODS One hundred twenty-eight subjects were included in this study. Atherosclerosis was defined as a maximum carotid plaque thickness (MCPT) of greater than 2 mm. A receiver operating characteristic curve analysis was used to determine the diagnostic value of serum CysC for atherosclerosis. The subjects were divided into two groups according to the CysC cut-off value. We screened for diabetes, hypertension, dyslipidemia, smoking status, alcohol consumption, and exercise behavior. The association between atherosclerosis and CysC levels was assessed using multivariate analysis. RESULTS The subjects were then divided into two groups according to the CysC cut-off value (0.73 mg/L). The median age of the high CysC group was 72 years (85% males), whereas that of the low CysC group was 61 years (63% males). The CysC levels were significantly correlated with Cr and estimated glomerular filtration rate (eGFR) values. Body-mass index, visceral fat area, hypertension, diabetes mellitus, and MCPT were significantly higher in the high CysC group than in the low CysC group. Furthermore, the eGFR was significantly lower in the high CysC group. Regarding lifestyle habits, only the exercise level was lower in the high CysC group than in the low CysC group. Multivariate analysis, adjusted for age and sex, revealed that high CysC levels were significantly associated with an MCPT of ≥ 2 mm (odds ratio: 2.92; 95%CI: 1.13-7.99). CONCLUSION Higher CysC levels were associated with an MCPT of ≥ 2 mm. The CysC cut-off value of 0.73 mg/L appears to aid in the diagnosis of atherosclerosis. PMID:28289532

  20. Adaptive user interfaces for relating high-level concepts to low-level photographic parameters

    NASA Astrophysics Data System (ADS)

    Scott, Edward; Madhawa Silva, Pubudu; Pardo, Bryan; Pappas, Thrasyvoulos N.

    2011-03-01

    Common controls for photographic editing can be difficult to use and have a significant learning curve. Often, a user does not know a direct mapping from a high-level concept (such as "soft") to the available parameters or controls. In addition, many concepts are subjective in nature, and the appropriate mapping may vary from user to user. To overcome these problems, we propose a system that can quickly learn a mapping from a high-level subjective concept onto low- level image controls using machine learning techniques. To learn such a concept, the system shows the user a series of training images that are generated by modifying a seed image along different dimensions (e.g., color, sharpness), and collects the user ratings of how well each training image matches the concept. Since it is known precisely how each modified example is different from the original, the system can determine the correlation between the user ratings and the image parameters to generate a controller tailored to the concept for the given user. The end result - a personalized image controller - is applicable to a variety of concepts. We have demonstrated the utility of this approach to relate low-level parameters, such as color balance and sharpness, to simple concepts, such as "lightness" and "crispness," as well as more complex and subjective concepts, such as "pleasantness." We have also applied the proposed approach to relate subband statistics (variance) to perceived roughness of visual textures (from the CUReT database).

  1. Radiative Lifetimes for High Levels of Neutral Fe

    NASA Astrophysics Data System (ADS)

    Lawler, James E.; Den Hartog, E.; Guzman, A.

    2013-01-01

    New radiative lifetime measurements for ~ 50 high lying levels of Fe I are reported. Laboratory astrophysics faces a challenge to provide basic spectroscopic data, especially reliable atomic transition probabilities, in the IR region for abundance studies. The availability of HgCdTe (HAWAII) detector arrays has opened IR spectral regions for extensive new spectroscopic studies. The SDSS III APOGEE project in the H-Band is an important example which will penetrate the dust obscuring the Galactic bulge. APOGEE will survey elemental abundances of 100,000 red giant stars in the bulge, bar, disk, and halo of the Milky Way. Many stellar spectra in the H-Band are, as expected, dominated by transitions of Fe I. Most of these IR transitions connect high levels of Fe. Our program has started an effort to meet this challenge with new radiative lifetime measurements on high lying levels of Fe I using time resolved laser induced fluorescence (TRLIF). The TRLIF method is typically accurate to 5% and is efficient. Our goal is to combine these accurate, absolute radiative lifetimes with emission branching fractions [1] to determine log(gf) values of the highest quality for Fe I lines in the UV, visible, and IR. This method was used very successfully by O’Brian et al. [2] on lower levels of Fe I. This method is still the best available for all but very simple spectra for which ab-initio theory is more accurate. Supported by NSF grant AST-0907732. [1] Branching fractions are being measured by M. Ruffoni and J. C. Pickering at Imperial College London. [2] O'Brian, T. R., Wickliffe, M. E., Lawler, J. E., Whaling, W., & Brault, J. W. 1991, J. Opt. Soc. Am. B 8, 1185

  2. ALICE: Project Overview and High Level Science Products

    NASA Astrophysics Data System (ADS)

    Soummer, Remi; Choquet, Elodie; Pueyo, Laurent; Brendan Hagan, J.; Gofas-Salas, Elena; Rajan, Abhijith; Perrin, Marshall D.; Chen, Christine; Debes, John H.; Golimowski, David A.; Hines, Dean C.; Schneider, Glenn; N'Diaye, Mamadou; Mawet, Dimitri; Marois, Christian; Barman, Travis

    2015-01-01

    We report on the status of the ALICE project (Archival Legacy Investigation of Circumstellar Environments), which consists in a consistent reanalysis of the entire HST-NICMOS coronagraphic archive. Over the last two years, we have developed a sophisticated pipeline able to handle the data of the 400 stars of the archive. This pipeline builds on the Karhunen-Loeve Image Projection (KLIP) algorithm, and was completed in the fall of 2014. We discuss the first processing and analysis results of the overall reduction campaign. As we will deliver high-level science products to the STScI MAST archive, we are defining a new standard format for high-contrast science products, which will be compatible with every new high-contrast imaging instrument (GPI, SPHERE, P1640, CHARIS, etc.) and used by the JWST coronagraphs. We present here the specifications of this standard.

  3. Exceptionally high levels of multiple mating in an army ant

    NASA Astrophysics Data System (ADS)

    Denny, A. Jay; Franks, Nigel R.; Powell, Scott; Edwards, Keith J.

    Most species of social insects have singly mated queens, although there are notable exceptions. Competing hypotheses have been proposed to explain the evolution of high levels of multiple mating, but this issue is far from resolved. Here we use microsatellites to investigate mating frequency in the army ant Eciton burchellii and show that queens mate with an exceptionally large number of males, eclipsing all but one other social insect species for which data are available. In addition we present evidence that suggests that mating is serial, continuing throughout the lifetime of the queen. This is the first demonstration of serial mating among social hymenoptera. We propose that high paternity within colonies is most likely to have evolved to increase genetic diversity and to counter high pathogen and parasite loads.

  4. High-level power analysis and optimization techniques

    NASA Astrophysics Data System (ADS)

    Raghunathan, Anand

    1997-12-01

    This thesis combines two ubiquitous trends in the VLSI design world--the move towards designing at higher levels of design abstraction, and the increasing importance of power consumption as a design metric. Power estimation and optimization tools are becoming an increasingly important part of design flows, driven by a variety of requirements such as prolonging battery life in portable computing and communication devices, thermal considerations and system cooling and packaging costs, reliability issues (e.g. electromigration, ground bounce, and I-R drops in the power network), and environmental concerns. This thesis presents a suite of techniques to automatically perform power analysis and optimization for designs at the architecture or register-transfer, and behavior or algorithm levels of the design hierarchy. High-level synthesis refers to the process of synthesizing, from an abstract behavioral description, a register-transfer implementation that satisfies the desired constraints. High-level synthesis tools typically perform one or more of the following tasks: transformations, module selection, clock selection, scheduling, and resource allocation and assignment (also called resource sharing or hardware sharing). High-level synthesis techniques for minimizing the area, maximizing the performance, and enhancing the testability of the synthesized designs have been investigated. This thesis presents high-level synthesis techniques that minimize power consumption in the synthesized data paths. This thesis investigates the effects of resource sharing on the power consumption in the data path, provides techniques to efficiently estimate power consumption during resource sharing, and resource sharing algorithms to minimize power consumption. The RTL circuit that is obtained from the high-level synthesis process can be further optimized for power by applying power-reducing RTL transformations. This thesis presents macro-modeling and estimation techniques for switching

  5. Space augmentation of military high-level waste disposal

    NASA Technical Reports Server (NTRS)

    English, T.; Lees, L.; Divita, E.

    1979-01-01

    Space disposal of selected components of military high-level waste (HLW) is considered. This disposal option offers the promise of eliminating the long-lived radionuclides in military HLW from the earth. A space mission which meets the dual requirements of long-term orbital stability and a maximum of one space shuttle launch per week over a period of 20-40 years, is a heliocentric orbit about halfway between the orbits of earth and Venus. Space disposal of high-level radioactive waste is characterized by long-term predictability and short-term uncertainties which must be reduced to acceptably low levels. For example, failure of either the Orbit Transfer Vehicle after leaving low earth orbit, or the storable propellant stage failure at perihelion would leave the nuclear waste package in an unplanned and potentially unstable orbit. Since potential earth reencounter and subsequent burn-up in the earth's atmosphere is unacceptable, a deep space rendezvous, docking, and retrieval capability must be developed.

  6. High level language-based robotic control system

    NASA Technical Reports Server (NTRS)

    Rodriguez, Guillermo (Inventor); Kreutz, Kenneth K. (Inventor); Jain, Abhinandan (Inventor)

    1996-01-01

    This invention is a robot control system based on a high level language implementing a spatial operator algebra. There are two high level languages included within the system. At the highest level, applications programs can be written in a robot-oriented applications language including broad operators such as MOVE and GRASP. The robot-oriented applications language statements are translated into statements in the spatial operator algebra language. Programming can also take place using the spatial operator algebra language. The statements in the spatial operator algebra language from either source are then translated into machine language statements for execution by a digital control computer. The system also includes the capability of executing the control code sequences in a simulation mode before actual execution to assure proper action at execution time. The robot's environment is checked as part of the process and dynamic reconfiguration is also possible. The languages and system allow the programming and control of multiple arms and the use of inward/outward spatial recursions in which every computational step can be related to a transformation from one point in the mechanical robot to another point to name two major advantages.

  7. High level language-based robotic control system

    NASA Technical Reports Server (NTRS)

    Rodriguez, Guillermo (Inventor); Kruetz, Kenneth K. (Inventor); Jain, Abhinandan (Inventor)

    1994-01-01

    This invention is a robot control system based on a high level language implementing a spatial operator algebra. There are two high level languages included within the system. At the highest level, applications programs can be written in a robot-oriented applications language including broad operators such as MOVE and GRASP. The robot-oriented applications language statements are translated into statements in the spatial operator algebra language. Programming can also take place using the spatial operator algebra language. The statements in the spatial operator algebra language from either source are then translated into machine language statements for execution by a digital control computer. The system also includes the capability of executing the control code sequences in a simulation mode before actual execution to assure proper action at execution time. The robot's environment is checked as part of the process and dynamic reconfiguration is also possible. The languages and system allow the programming and control of multiple arms and the use of inward/outward spatial recursions in which every computational step can be related to a transformation from one point in the mechanical robot to another point to name two major advantages.

  8. Characterization of Transducers and Resonators under High Drive Levels

    NASA Technical Reports Server (NTRS)

    Sherrit, Stewart; Bao, X.; Sigel, D. A.; Gradziel, M. J.; Askins, S. A.; Dolgin, B. P.; Bar-Cohen, Y.

    2001-01-01

    In many applications, piezoelectric transducers are driven at AC voltage levels well beyond the level for which the material was nominally characterized. In this paper we describe an experimental setup that allows for the determination of the main transducer or resonator properties under large AC drive. A sinusoidal voltage from a waveform generator is amplified and applied across the transducer/resonator in series with a known high power resistor. The amplitude of applied voltage and the amplitude and the relative phase of the current through the resistor are monitored on a digital scope. The frequency of the applied signal is swept through resonance and the voltage/current signals are recorded. After corrections for the series resistance and parasitic elements the technique allows for the determination of the complex impedance spectra of the sample as a function of frequency. In addition, access to the current signal allows for the direct investigation of non-linear effects through the application of Fourier transform techniques on the current signal. Our results indicate that care is required when interpreting impedance data at high drive level due to the frequency dependence of the dissipated power. Although the transducer/resonator at a single frequency and after many cycles may reach thermal equilibrium, the spectra as a whole cannot be considered an isothermal measurement due to the temperature change with frequency. Methods to correct for this effect will be discussed. Results determined from resonators of both soft and hard PZT and a ultrasonic horn transducer are presented.

  9. Permitting plan for the high-level waste interim storage

    SciTech Connect

    Deffenbaugh, M.L.

    1997-04-23

    This document addresses the environmental permitting requirements for the transportation and interim storage of solidified high-level waste (HLW) produced during Phase 1 of the Hanford Site privatization effort. Solidified HLW consists of canisters containing vitrified HLW (glass) and containers that hold cesium separated during low-level waste pretreatment. The glass canisters and cesium containers will be transported to the Canister Storage Building (CSB) in a U.S. Department of Energy (DOE)-provided transportation cask via diesel-powered tractor trailer. Tri-Party Agreement (TPA) Milestone M-90 establishes a new major milestone, and associated interim milestones and target dates, governing acquisition and/or modification of facilities necessary for: (1) interim storage of Tank Waste Remediation Systems (TWRS) immobilized HLW (IHLW) and other canistered high-level waste forms; and (2) interim storage and disposal of TWRS immobilized low-activity tank waste (ILAW). An environmental requirements checklist and narrative was developed to identify the permitting path forward for the HLW interim storage (HLWIS) project (See Appendix B). This permitting plan will follow the permitting logic developed in that checklist.

  10. Prostate Cancer, High Cortisol Levels and Complex Hormonal Interaction.

    PubMed

    Fabre, Bibiana; Grosman, Halina; Gonzalez, Diego; Machulsky, Nahuel Fernandez; Repetto, Esteban M; Mesch, Viviana; Lopez, Miguel Angel; Mazza, Osvaldo; Berg, Gabriela

    2016-01-01

    Prostate cancer (PCa) is one of the most common diseases in men. It is important to assess prognostic factors and whether high cortisol levels and complex hormonal interactions could be responsible for PCa development. We evaluated the relationship between cortisol, leptin and estrogens in 141 men, 71 with PCa and the remaining 70 constituting a low risk group (LRG). They were recruited for this study from a total of 2906 middleaged men (ages 4570 years) who completed an evaluation for prostatic diseases at the Urology Division, Hospital de Clinicas "Jose de San Martin", University of Buenos Aires, in May 2009. In this cross sectional study, cortisol, PSA, totaltestosterone, freetestosterone, bioavailable testosterone, LH and estradiol were measured in serum. We observed increased cortisol levels in PCa patients as compared to LRG cases (p=0.004,). Leptin and estradiol levels were also higher in PCa patients (p=0.048; p<0.0001, respectively). Logistic regression analysis indicated that serum cortisol (OR: 1.110 (95% CI 1.0161.213), p=0.022), estradiol (OR: 1.044 (95% CI 1.0081.081), p=0.016) and leptin (OR: 1.248 (95% CI 1.0481.487), p=0.013) explained 27% of the variance of dependent variables, even after adjusting for age, smoking, BMI and waist circumference. We found increased cortisol levels in PCa patients as compared to LRG, as well as an altered circulating hormonal profile.

  11. Effects of high vs low-level radiation exposure

    SciTech Connect

    Bond, V.P.

    1983-01-01

    In order to appreciate adequately the various possible effects of radiation, particularly from high-level vs low-level radiation exposure (HLRE, vs LLRE), it is necessary to understand the substantial differences between (a) exposure as used in exposure-incidence curves, which are always initially linear and without threshold, and (b) dose as used in dose-response curves, which always have a threshold, above which the function is curvilinear with increasing slope. The differences are discussed first in terms of generally familiar nonradiation situations involving dose vs exposure, and then specifically in terms of exposure to radiation, vs a dose of radiation. Examples are given of relevant biomedical findings illustrating that, while dose can be used with HLRE, it is inappropriate and misleading the LLRE where exposure is the conceptually correct measure of the amount of radiation involved.

  12. University-Level Research Projects for High School Students

    NASA Technical Reports Server (NTRS)

    McConnell, Mark L.

    2000-01-01

    The goal of this project was to provide an opportunity for high school students to participate in university-level research projects. In this case, students from Pinkerton Academy (Derry, New Hampshire) were invited to participate in efforts to catalog data from the COMPTEL experiment on NASA's Compton Gamma-Ray Observatory (CGRO). These activities were part of a senior level honors course at Pinkerton. Although the success of this particular program was rather limited, we feel that the general concept is a sound one. In principle, the concept of partnerships between local schools and university researchers is one that could be especially attractive to soft money researchers. Programs can be carefully designed to benefit both the students and the research program.

  13. Salivary Fluoride Levels after Use of High-Fluoride Dentifrice

    PubMed Central

    Vale, Glauber Campos; Cruz, Priscila Figueiredo; Bohn, Ana Clarissa Cavalcante Elvas; de Moura, Marcoeli Silva

    2015-01-01

    The aim of the study was to evaluate salivary fluoride (F) availability after toothbrushing with a high-F dentifrice. Twelve adult volunteers took part in this crossover and blind study. F concentration in saliva was determined after brushing with a high-F dentifrice (5000 µg F/g) or with a conventional F concentration dentifrice (1100 µg F/g) followed by a 15 mL distilled water rinse. Samples of nonstimulated saliva were collected on the following times: before (baseline), and immediately after spit (time = 0) and after 1, 2, 3, 4, 5, 10, 15, 20, 30, 45, 60, 90, and 120 min. F analysis was performed with a fluoride-sensitive electrode and the area under curve of F salivary concentration × time (µg F/mL × min−1) was calculated. At baseline, no significant difference was found among dentifrices (P > 0.05). After brushing, both dentifrices caused an elevated fluoride level in saliva; however salivary F concentration was significantly higher at all times, when high-F dentifrice was used (P < 0.01). Even after 120 min, salivary F concentration was still higher than the baseline values for both dentifrices (P < 0.001). High-F dentifrice enhanced the bioavailability of salivary F, being an option for caries management in patients with high caries risk. PMID:25821849

  14. Salivary fluoride levels after use of high-fluoride dentifrice.

    PubMed

    Vale, Glauber Campos; Cruz, Priscila Figueiredo; Bohn, Ana Clarissa Cavalcante Elvas; de Moura, Marcoeli Silva

    2015-01-01

    The aim of the study was to evaluate salivary fluoride (F) availability after toothbrushing with a high-F dentifrice. Twelve adult volunteers took part in this crossover and blind study. F concentration in saliva was determined after brushing with a high-F dentifrice (5000 µg F/g) or with a conventional F concentration dentifrice (1100 µg F/g) followed by a 15 mL distilled water rinse. Samples of nonstimulated saliva were collected on the following times: before (baseline), and immediately after spit (time = 0) and after 1, 2, 3, 4, 5, 10, 15, 20, 30, 45, 60, 90, and 120 min. F analysis was performed with a fluoride-sensitive electrode and the area under curve of F salivary concentration × time (µg F/mL × min(-1)) was calculated. At baseline, no significant difference was found among dentifrices (P > 0.05). After brushing, both dentifrices caused an elevated fluoride level in saliva; however salivary F concentration was significantly higher at all times, when high-F dentifrice was used (P < 0.01). Even after 120 min, salivary F concentration was still higher than the baseline values for both dentifrices (P < 0.001). High-F dentifrice enhanced the bioavailability of salivary F, being an option for caries management in patients with high caries risk.

  15. Hip Arthroscopy in High-Level Baseball Players.

    PubMed

    Byrd, J W Thomas; Jones, Kay S

    2015-08-01

    To report the results of hip arthroscopy among high-level baseball players as recorded by outcome scores and return to baseball. All patients undergoing hip arthroscopy were prospectively assessed with the modified Harris Hip Score. On review of all procedures performed over a 12-year period, 44 hips were identified among 41 intercollegiate or professional baseball players who had achieved 2-year follow-up. Among the 41 players, follow-up averaged 45 months (range, 24 to 120 months), with a mean age of 23 years (range, 18 to 34 years). There were 23 collegiate (1 bilateral) and 18 professional (2 bilateral) baseball players, including 10 Major League Baseball players. Of the 8 Major League Baseball pitchers, 6 (75%) also underwent ulnar collateral ligament elbow surgery. Improvement in the modified Harris Hip Score averaged 13 points (from 81 points preoperatively to 94 points postoperatively); a paired-samples t test determined that this mean improvement of 13 points was statistically significant (P < .001). Players returned to baseball after 42 of 44 procedures (95%) at a mean of 4.3 months (range, 3 to 8 months), with 90% regaining the ability to participate at their previous level of competition. There were no complications. Three players (1 bilateral) underwent repeat arthroscopy. This study supports the idea that arthroscopic treatment for a variety of hip pathologies in high-level baseball players provides a successful return to sport and improvement in functional outcome scores. Level IV, therapeutic case series. Copyright © 2015 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.

  16. Defense High Level Waste Disposal Container System Description Document

    SciTech Connect

    N. E. Pettit

    2001-07-13

    The Defense High Level Waste Disposal Container System supports the confinement and isolation of waste within the Engineered Barrier System of the Monitored Geologic Repository (MGR). Disposal containers are loaded and sealed in the surface waste handling facilities, transferred to the underground through the accesses using a rail mounted transporter, and emplaced in emplacement drifts. The defense high level waste (HLW) disposal container provides long-term confinement of the commercial HLW and defense HLW (including immobilized plutonium waste forms [IPWF]) placed within disposable canisters, and withstands the loading, transfer, emplacement, and retrieval loads and environments. US Department of Energy (DOE)-owned spent nuclear fuel (SNF) in disposable canisters may also be placed in a defense HLW disposal container along with commercial HLW waste forms, which is known as co-disposal. The Defense High Level Waste Disposal Container System provides containment of waste for a designated period of time, and limits radionuclide release. The disposal container/waste package maintains the waste in a designated configuration, withstands maximum handling and rockfall loads, limits the individual canister temperatures after emplacement, resists corrosion in the expected handling and repository environments, and provides containment of waste in the event of an accident. Defense HLW disposal containers for HLW disposal will hold up to five HLW canisters. Defense HLW disposal containers for co-disposal will hold up to five HLW canisters arranged in a ring and one DOE SNF canister inserted in the center and/or one or more DOE SNF canisters displacing a HLW canister in the ring. Defense HLW disposal containers also will hold two Multi-Canister Overpacks (MCOs) and two HLW canisters in one disposal container. The disposal container will include outer and inner cylinders, outer and inner cylinder lids, and may include a canister guide. An exterior label will provide a means by

  17. Development of a High Level Waste Tank Inspection System

    SciTech Connect

    Appel, D.K.; Loibl, M.W.; Meese, D.C.

    1995-03-21

    The Westinghouse Savannah River Technology Center was requested by it`s sister site, West Valley Nuclear Service (WVNS), to develop a remote inspection system to gather wall thickness readings of their High Level Waste Tanks. WVNS management chose to take a proactive approach to gain current information on two tanks t hat had been in service since the early 70`s. The tanks contain high level waste, are buried underground, and have only two access ports to an annular space between the tank and the secondary concrete vault. A specialized remote system was proposed to provide both a visual surveillance and ultrasonic thickness measurements of the tank walls. A magnetic wheeled crawler was the basis for the remote delivery system integrated with an off-the-shelf Ultrasonic Data Acquisition System. A development program was initiated for Savannah River Technology Center (SRTC) to design, fabricate, and test a remote system based on the Crawler. The system was completed and involved three crawlers to perform the needed tasks, an Ultrasonic Crawler, a Camera Crawler, and a Surface Prep Crawler. The crawlers were computer controlled so that their operation could be done remotely and their position on the wall could be tracked. The Ultrasonic Crawler controls were interfaced with ABB Amdata`s I-PC, Ultrasonic Data Acquisition System so that thickness mapping of the wall could be obtained. A second system was requested by Westinghouse Savannah River Company (WSRC), to perform just ultrasonic mapping on their similar Waste Storage Tanks; however, the system needed to be interfaced with the P-scan Ultrasonic Data Acquisition System. Both remote inspection systems were completed 9/94. Qualifications tests were conducted by WVNS prior to implementation on the actual tank and tank development was achieved 10/94. The second inspection system was deployed at WSRC 11/94 with success, and the system is now in continuous service inspecting the remaining high level waste tanks at WSRC.

  18. CEMENTITIOUS GROUT FOR CLOSING SRS HIGH LEVEL WASTE TANKS - #12315

    SciTech Connect

    Langton, C.; Burns, H.; Stefanko, D.

    2012-01-10

    In 1997, the first two United States Department of Energy (US DOE) high level waste tanks (Tanks 17-F and 20-F: Type IV, single shell tanks) were taken out of service (permanently closed) at the Savannah River Site (SRS). In 2012, the DOE plans to remove from service two additional Savannah River Site (SRS) Type IV high-level waste tanks, Tanks 18-F and 19-F. These tanks were constructed in the late 1950's and received low-heat waste and do not contain cooling coils. Operational closure of Tanks 18-F and 19-F is intended to be consistent with the applicable requirements of the Resource Conservation and Recovery Act (RCRA) and the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) and will be performed in accordance with South Carolina Department of Health and Environmental Control (SCDHEC). The closure will physically stabilize two 4.92E+04 cubic meter (1.3 E+06 gallon) carbon steel tanks and isolate and stabilize any residual contaminants left in the tanks. The closure will also fill, physically stabilize and isolate ancillary equipment abandoned in the tanks. A Performance Assessment (PA) has been developed to assess the long-term fate and transport of residual contamination in the environment resulting from the operational closure of the F-Area Tank Farm (FTF) waste tanks. Next generation flowable, zero-bleed cementitious grouts were designed, tested, and specified for closing Tanks 18-F and 19-F and for filling the abandoned equipment. Fill requirements were developed for both the tank and equipment grouts. All grout formulations were required to be alkaline with a pH of 12.4 and chemically reduction potential (Eh) of -200 to -400 to stabilize selected potential contaminants of concern. This was achieved by including Portland cement and Grade 100 slag in the mixes, respectively. Ingredients and proportions of cementitious reagents were selected and adjusted, respectively, to support the mass placement strategy developed by closure

  19. Stagnation Region Heat Transfer Augmentation at Very High Turbulence Levels

    SciTech Connect

    Ames, Forrest; Kingery, Joseph E.

    2015-06-17

    A database for stagnation region heat transfer has been extended to include heat transfer measurements acquired downstream from a new high intensity turbulence generator. This work was motivated by gas turbine industry heat transfer designers who deal with heat transfer environments with increasing Reynolds numbers and very high turbulence levels. The new mock aero-combustor turbulence generator produces turbulence levels which average 17.4%, which is 37% higher than the older turbulence generator. The increased level of turbulence is caused by the reduced contraction ratio from the liner to the exit. Heat transfer measurements were acquired on two large cylindrical leading edge test surfaces having a four to one range in leading edge diameter (40.64 cm and 10.16 cm). Gandvarapu and Ames [1] previously acquired heat transfer measurements for six turbulence conditions including three grid conditions, two lower turbulence aero-combustor conditions, and a low turbulence condition. The data are documented and tabulated for an eight to one range in Reynolds numbers for each test surface with Reynolds numbers ranging from 62,500 to 500,000 for the large leading edge and 15,625 to 125,000 for the smaller leading edge. The data show augmentation levels of up to 136% in the stagnation region for the large leading edge. This heat transfer rate is an increase over the previous aero-combustor turbulence generator which had augmentation levels up to 110%. Note, the rate of increase in heat transfer augmentation decreases for the large cylindrical leading edge inferring only a limited level of turbulence intensification in the stagnation region. The smaller cylindrical leading edge shows more consistency with earlier stagnation region heat transfer results correlated on the TRL (Turbulence, Reynolds number, Length scale) parameter. The downstream regions of both test surfaces continue to accelerate the flow but at a much lower rate than the leading edge. Bypass transition occurs

  20. Modern Alchemy: Solidifying high-level nuclear waste

    SciTech Connect

    Newton, C.C.

    1997-07-01

    The U.S. Department of Energy is putting a modern version of alchemy to work to produce an answer to a decades-old problem. It is taking place at the Savannah River Site (SRS) in Aiken, South Carolina and at the West Valley Demonstration Project (WVDP) near Buffalo, New York. At both locations, contractor Westinghouse Electric Corporation is applying technology that is turning liquid high-level radioactive waste (HLW) into a stabilized, durable glass for safer and easier management. The process is called vitrification. SRS and WVDP are now operating the nation`s first full-scale HLW vitrification plants.

  1. Corrosion and failure processes in high-level waste tanks

    SciTech Connect

    Mahidhara, R.K.; Elleman, T.S.; Murty, K.L.

    1992-11-01

    A large amount of radioactive waste has been stored safely at the Savannah River and Hanford sites over the past 46 years. The aim of this report is to review the experimental corrosion studies at Savannah River and Hanford with the intention of identifying the types and rates of corrosion encountered and indicate how these data contribute to tank failure predictions. The compositions of the High-Level Wastes, mild steels used in the construction of the waste tanks and degradation-modes particularly stress corrosion cracking and pitting are discussed. Current concerns at the Hanford Site are highlighted.

  2. High level trigger online calibration framework in ALICE

    NASA Astrophysics Data System (ADS)

    Bablok, S. R.; Djuvsland, Ø.; Kanaki, K.; Nystrand, J.; Richter, M.; Röhrich, D.; Skjerdal, K.; Ullaland, K.; Øvrebekk, G.; Larsen, D.; Alme, J.; Alt, T.; Lindenstruth, V.; Steinbeck, T. M.; Thäder, J.; Kebschull, U.; Böttger, S.; Kalcher, S.; Lara, C.; Panse, R.; Appelshäuser, H.; Ploskon, M.; Helstrup, H.; Hetland, K. F.; Haaland, Ø.; Roed, K.; Thingnæs, T.; Aamodt, K.; Hille, P. T.; Lovhoiden, G.; Skaali, B.; Tveter, T.; Das, I.; Chattopadhyay, S.; Becker, B.; Cicalo, C.; Marras, D.; Siddhanta, S.; Cleymans, J.; Szostak, A.; Fearick, R.; Vaux, G. d.; Vilakazi, Z.

    2008-07-01

    The ALICE High Level Trigger (HLT) is designed to perform event analysis of heavy ion and proton-proton collisions as well as calibration calculations online. A large PC farm, currently under installation, enables analysis algorithms to process these computationally intensive tasks. The HLT receives event data from all major detectors in ALICE. Interfaces to the various other systems provide the analysis software with required additional information. Processed results are sent back to the corresponding systems. To allow online performance monitoring of the detectors an interface for visualizing these results has been developed.

  3. THE AMERICAN HIGH SCHOOL GRADUATION RATE: TRENDS AND LEVELS*

    PubMed Central

    Heckman, James J.; LaFontaine, Paul A.

    2009-01-01

    This paper applies a unified methodology to multiple data sets to estimate both the levels and trends in U.S. high school graduation rates. We establish that (a) the true rate is substantially lower than widely used measures; (b) it peaked in the early 1970s; (c) majority/minority differentials are substantial and have not converged for 35 years; (d) lower post-1970 rates are not solely due to increasing immigrant and minority populations; (e) our findings explain part of the slowdown in college attendance and rising college wage premiums; and (f) widening graduation differentials by gender help explain increasing male-female college attendance gaps. PMID:20625528

  4. Smoking as a determinant of high organochlorine levels in Greenland.

    PubMed

    Deutch, Bente; Pedersen, Henning Sloth; Jørgensen, Eva C Bonefeld; Hansen, Jens C

    2003-01-01

    The authors investigated the accumulation of organochlorines among smoking and nonsmoking Inuit hunters (n = 48) in Uummanaq, Greenland, a population with high dietary exposure to persistent organic pollutants (POPs). Human plasma organochlorine levels were positively correlated with age, marine diet, and smoking or plasma cotinine in multiple linear-regression models (p < 0.001). Body mass index was inversely correlated with organochlorine accumulation, independent of smoking status. These findings confirm that the source of POPs among the Inuit in Greenland is diet, but smoking is an important determinant of POP bioaccumulation. Smoking cessation may provide a means to lower the body burden of POPs.

  5. High-level neutron coincidence counter maintenance manual

    SciTech Connect

    Swansen, J.; Collinsworth, P.

    1983-05-01

    High-level neutron coincidence counter operational (field) calibration and usage is well known. This manual makes explicit basic (shop) check-out, calibration, and testing of new units and is a guide for repair of failed in-service units. Operational criteria for the major electronic functions are detailed, as are adjustments and calibration procedures, and recurrent mechanical/electromechanical problems are addressed. Some system tests are included for quality assurance. Data on nonstandard large-scale integrated (circuit) components and a schematic set are also included.

  6. Market Designs for High Levels of Variable Generation: Preprint

    SciTech Connect

    Milligan, M.; Holttinen, H.; Kiviluoma, J.; Orths, A.; Lynch, M.; Soder, L.

    2014-10-01

    Variable renewable generation is increasing in penetration in modern power systems, leading to higher variability in the supply and price of electricity as well as lower average spot prices. This raises new challenges, particularly in ensuring sufficient capacity and flexibility from conventional technologies. Because the fixed costs and lifetimes of electricity generation investments are significant, designing markets and regulations that ensure the efficient integration of renewable generation is a significant challenge. This papers reviews the state of play of market designs for high levels of variable generation in the United States and Europe and considers new developments in both regions.

  7. High resolution measurement of water levels in optical components

    NASA Astrophysics Data System (ADS)

    Murrieta-Rico, Fabian N.; Petranovskii, Vitalii; Sergiyenko, Oleg; Hernandez-Balbuena, Daniel; Raymond-Herrera, Oscar

    2016-09-01

    Systems for optical analysis use vacuum chambers, where low pressures are reached. Remaining water molecules are the prevalent contaminant in high vacuum chambers. For this reason measurement of water levels is an important task that allows correct equipment operation. In this work, a different approach is presented for detecting and quantifying the water molecules inside a the vacuum chamber used in optical systems. A zeolite coated quartz crystal microbalance is used for detecting the water molecules, and the change in the resonance frequency is measured using a novel technique known as the principle of rational approximations. Theoretical results show how nanograms of adsorbed molecules are measured, and the number of molecules are quantified.

  8. High-level wastes: DOE names three sites for characterization

    SciTech Connect

    1986-07-01

    DOE announced in May 1986 that there will be there site characterization studies made to determine suitability for a high-level radioactive waste repository. The studies will include several test drillings to the proposed disposal depths. Yucca Mountain, Nevada; Deaf Smith Country, Texas, and Hanford, Washington were identified as the study sites, and further studies for a second repository site in the East were postponed. The affected states all filed suits in federal circuit courts because they were given no advance warning of the announcement of their selection or the decision to suspend work on a second repository. Criticisms of the selection process include the narrowing or DOE options.

  9. Elevated glucose levels in early puerperium, and association with high cortisol levels during parturition.

    PubMed

    Risberg, Anitha; Sjöquist, Mats; Wedenberg, Kaj; Larsson, Anders

    2016-07-01

    Background Gestational diabetes is one of the commonest metabolic problems associated with pregnancy and an accurate diagnosis is critical for the care. Research has shown that pregnant women have high levels of cortisol during the last stage of parturition. As cortisol is a diabetogenic hormone causing increased glucose levels, we wanted to study the association between cortisol and glucose levels during parturition. Materials and methods Glucose and cortisol were analyzed during parturition in 50 females divided according to slow (n = 11) and normal labors (n = 39). Blood samples were analyzed three times during the parturition and four times in the first day after delivery. Glucose levels were also measured once in each trimester. Results In the normal group, the glucose concentration increased from 6.2 (IQR 5.6-8.0) mmol/L in the latency phase to 11.6 (10.0-13.3) mmol/L at aftercare (p < 0.05). After parturition the glucose concentrations decreased gradually. There were significant Spearman rank correlations between glucose and cortisol values. Conclusions The changes associated with birth cause significant elevations of cortisol and glucose around parturition.

  10. Abnormally high serum ferritin levels among professional road cyclists

    PubMed Central

    Zotter, H; Robinson, N; Zorzoli, M; Schattenberg, L; Saugy, M; Mangin, P

    2004-01-01

    Background: An international, longitudinal medical follow up examination of male professional road cyclists revealed excessively elevated serum ferritin levels. Objective: To evaluate the importance of elevated ferritin values among professional cyclists, their relationship with age and nationality, and their evolution over 3 years. Methods: Over 1000 serum ferritin values were collected. Other parameters were included in order to exclude conditions which might have increased ferritin levels without changing body iron stores. Results: In 1999, over 45% of riders displayed ferritin values above 300 ng/ml and one fourth levels over 500 ng/ml. These percentages had decreased to 27% and 9%, respectively, 3 years later, while the overall average, which was above the normal limits in 1999, had decreased by 33% in 3 years. Older cyclists had higher ferritin values than younger cyclists. There was also a relationship between ferritin levels and the nationality of the cyclists. Analysis of 714 riders in 2000 and 2002 showed only a slight and insignificant decrease in the mean ferritin value although those with initially elevated iron stores had a much greater decrease. Conclusion: Professional road cyclists used excessive iron supplementation leading to high serum ferritin levels correlating with increased body iron stores. Although the situation progressively improved over 3 years, it remains worrying as increased body iron stores are related to health complications. Therefore, prevention in addition to the fight against doping should be a main goal of the UCI. Aggressive therapy for athletes with excessive ferritin values should be carried out at or before the end of their careers. PMID:15562163

  11. HIGH LEVELS OF URANIUM IN GROUNDWATER OF ULAANBAATAR, MONGOLIA

    PubMed Central

    Nriagu, Jerome; Nam, Dong-Ha; Ayanwola, Titilayo A.; Dinh, Hau; Erdenechimeg, Erdenebayar; Ochir, Chimedsuren; Bolormaa, Tsend-Ayush

    2011-01-01

    Water samples collected from 129 wells in seven of the nine sub-divisions of Ulaanbaatar were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) using Clean Lab methods. The levels of many trace elements were found to be very low with the average concentrations (ranges in brackets) being 0.9 (<0.1-7.9) μg/L for As; 7.7 (0.12-177) μg/L for Mn; 0.2 (<0.05-1.9) μg/L for Co; 16 (<0.1-686) μg/L for Zn; 0.7 (<0.1-1.8) μg/L for Se; <0.1 (<0.02-0.69) μg/L for Cd; and 1.3 (<0.02-32) μg/L for Pb. The levels of uranium were surprisingly elevated (mean, 4.6 μg/L; range <0.01-57 μg/L, with the values for many samples exceeding the World Health Organization's guideline of 15 μg/L for uranium in drinking water. Local rocks and soils appear to be the natural source of the uranium. The levels of uranium in Ulaanbaatar's groundwater are in the range that has been associated with nephrotoxicity, high blood pressure, bone dysfunction and likely reproductive impairment in human populations. We consider the risk associated with drinking the groundwater with elevated levels of uranium in Ulaanbaatar to be a matter for some public health concern and conclude that the paucity of data on chronic effects of low level exposure is a risk factor for continuing the injury to many people in this city. PMID:22142646

  12. Online Pattern Recognition for the ALICE High Level Trigger

    NASA Astrophysics Data System (ADS)

    Lindenstruth, V.; Loizides, C.; Rohrich, D.; Skaali, B.; Steinbeck, T.; Stock, R.; Tilsner, H.; Ullaland, K.; Vestbo, A.; Vik, T.

    2004-06-01

    The ALICE high level trigger has to process data online, in order to select interesting (sub)events, or to compress data efficiently by modeling techniques. Focusing on the main data source, the time projection chamber (TPC), we present two pattern recognition methods under investigation: a sequential approach (cluster finder and track follower) and an iterative approach (track candidate finder and cluster deconvoluter). We show, that the former is suited for pp and low multiplicity PbPb collisions, whereas the latter might be applicable for high multiplicity PbPb collisions of dN/dy>3000. Based on the developed tracking schemes we show that using modeling techniques, a compression factor of around 10 might be achievable.

  13. Remote ignitability analysis of high-level radioactive waste

    SciTech Connect

    Lundholm, C.W.; Morgan, J.M.; Shurtliff, R.M.; Trejo, L.E.

    1992-09-01

    The Idaho Chemical Processing Plant (ICPP), was used to reprocess nuclear fuel from government owned reactors to recover the unused uranium-235. These processes generated highly radioactive liquid wastes which are stored in large underground tanks prior to being calcined into a granular solid. The Resource Conservation and Recovery Act (RCRA) and state/federal clean air statutes require waste characterization of these high level radioactive wastes for regulatory permitting and waste treatment purposes. The determination of the characteristic of ignitability is part of the required analyses prior to calcination and waste treatment. To perform this analysis in a radiologically safe manner, a remoted instrument was needed. The remote ignitability Method and Instrument will meet the 60 deg. C. requirement as prescribed for the ignitability in method 1020 of SW-846. The method for remote use will be equivalent to method 1020 of SW-846.

  14. Psilocybin impairs high-level but not low-level motion perception.

    PubMed

    Carter, Olivia L; Pettigrew, John D; Burr, David C; Alais, David; Hasler, Felix; Vollenweider, Franz X

    2004-08-26

    The hallucinogenic serotonin(1A&2A) agonist psilocybin is known for its ability to induce illusions of motion in otherwise stationary objects or textured surfaces. This study investigated the effect of psilocybin on local and global motion processing in nine human volunteers. Using a forced choice direction of motion discrimination task we show that psilocybin selectively impairs coherence sensitivity for random dot patterns, likely mediated by high-level global motion detectors, but not contrast sensitivity for drifting gratings, believed to be mediated by low-level detectors. These results are in line with those observed within schizophrenic populations and are discussed in respect to the proposition that psilocybin may provide a model to investigate clinical psychosis and the pharmacological underpinnings of visual perception in normal populations.

  15. High-level waste melter alternatives assessment report

    SciTech Connect

    Calmus, R.B.

    1995-02-01

    This document describes the Tank Waste Remediation System (TWRS) High-Level Waste (HLW) Program`s (hereafter referred to as HLW Program) Melter Candidate Assessment Activity performed in fiscal year (FY) 1994. The mission of the TWRS Program is to store, treat, and immobilize highly radioactive Hanford Site waste (current and future tank waste and encapsulated strontium and cesium isotopic sources) in an environmentally sound, safe, and cost-effective manner. The goal of the HLW Program is to immobilize the HLW fraction of pretreated tank waste into a vitrified product suitable for interim onsite storage and eventual offsite disposal at a geologic repository. Preparation of the encapsulated strontium and cesium isotopic sources for final disposal is also included in the HLW Program. As a result of trade studies performed in 1992 and 1993, processes planned for pretreatment of tank wastes were modified substantially because of increasing estimates of the quantity of high-level and transuranic tank waste remaining after pretreatment. This resulted in substantial increases in needed vitrification plant capacity compared to the capacity of original Hanford Waste Vitrification Plant (HWVP). The required capacity has not been finalized, but is expected to be four to eight times that of the HWVP design. The increased capacity requirements for the HLW vitrification plant`s melter prompted the assessment of candidate high-capacity HLW melter technologies to determine the most viable candidates and the required development and testing (D and T) focus required to select the Hanford Site HLW vitrification plant melter system. An assessment process was developed in early 1994. This document describes the assessment team, roles of team members, the phased assessment process and results, resulting recommendations, and the implementation strategy.

  16. Wind resource quality affected by high levels of renewables

    DOE PAGES

    Diakov, Victor

    2015-06-17

    For solar photovoltaic (PV) and wind resources, the capacity factor is an important parameter describing the quality of the resource. As the share of variable renewable resources (such as PV and wind) on the electric system is increasing, so does curtailment (and the fraction of time when it cannot be avoided). At high levels of renewable generation, curtailments effectively change the practical measure of resource quality from capacity factor to the incremental capacity factor. The latter accounts only for generation during hours of no curtailment and is directly connected with the marginal capital cost of renewable generators for a givenmore » level of renewable generation during the year. The Western U.S. wind generation is analyzed hourly for a system with 75% of annual generation from wind, and it is found that the value for the system of resources with equal capacity factors can vary by a factor of 2, which highlights the importance of using the incremental capacity factor instead. Finally, the effect is expected to be more pronounced in smaller geographic areas (or when transmission limitations imposed) and less pronounced at lower levels of renewable energy in the system with less curtailment.« less

  17. Wind resource quality affected by high levels of renewables

    SciTech Connect

    Diakov, Victor

    2015-06-17

    For solar photovoltaic (PV) and wind resources, the capacity factor is an important parameter describing the quality of the resource. As the share of variable renewable resources (such as PV and wind) on the electric system is increasing, so does curtailment (and the fraction of time when it cannot be avoided). At high levels of renewable generation, curtailments effectively change the practical measure of resource quality from capacity factor to the incremental capacity factor. The latter accounts only for generation during hours of no curtailment and is directly connected with the marginal capital cost of renewable generators for a given level of renewable generation during the year. The Western U.S. wind generation is analyzed hourly for a system with 75% of annual generation from wind, and it is found that the value for the system of resources with equal capacity factors can vary by a factor of 2, which highlights the importance of using the incremental capacity factor instead. Finally, the effect is expected to be more pronounced in smaller geographic areas (or when transmission limitations imposed) and less pronounced at lower levels of renewable energy in the system with less curtailment.

  18. High-spin level scheme of doubly odd 128I

    NASA Astrophysics Data System (ADS)

    Ding, B.; Wang, H. X.; Jiang, H.; Zhang, Y. H.; Zhou, X. H.; Zhao, Y. M.; Wang, S. T.; Liu, M. L.; Li, G. S.; Zheng, Y.; Zhang, N. T.; Zhou, H. B.; Ma, Y. J.; Sasakiz, Y.; Yamada, K.; Ohshima, H.; Yokose, S.; Ishizuka, M.; Komatsubara, T.; Furuno, K.

    2012-09-01

    High-spin states in the odd-odd 128I have been studied experimentally using the 124Sn(7Li, 3nγ)128I reaction at beam energies of 28 and 32 MeV. A level scheme built on the known T1/2=175 ns, (6-) isomer has been established. The low-lying states in the level scheme have been proposed to be associated with the πd5/2⊗νh11/2 and πg7/2⊗νh11/2 two-quasiparticle excitations. The strongly populated negative-parity states, forming a bandlike structure, have been assigned to be based on the πg7/2⊗νh11/2 configuration. The πh11/2⊗νh11/2 multiplet 9+, 10+, and 11+ members have been identified according to the systematics of the corresponding level structures observed in the lighter odd-odd iodine isotopes. The two-quasiparticle configuration assignments are supported by the nucleon pair approximation and empirical shell-model calculations.

  19. Pupil responses to high-level image content.

    PubMed

    Naber, Marnix; Nakayama, Ken

    2013-05-17

    The link between arousal and pupil dilation is well studied, but it is less known that other cognitive processes can trigger pupil responses. Here we present evidence that pupil responses can be induced by high-level scene processing, independent of changes in low-level features or arousal. In Experiment 1, we recorded changes in pupil diameter of observers while they viewed a variety of natural scenes with or without a sun that were presented either upright or inverted. Image inversion had the strongest effect on the pupil responses. The pupil constricted more to the onset of upright images as compared to inverted images. Furthermore, the amplitudes of pupil constrictions to viewing images containing a sun were larger relative to control images. In Experiment 2, we presented cartoon versions of upright and inverted pictures that included either a sun or a moon. The image backgrounds were kept identical across conditions. Similar to Experiment 1, upright images triggered pupil constrictions with larger amplitudes than inverted images and images of the sun evoked greater pupil contraction than images of the moon. We suggest that the modulations of pupil responses were due to higher-level interpretations of image content.

  20. ATW system impact on high-level waste

    SciTech Connect

    Arthur, E.D.

    1992-12-01

    This report discusses the Accelerator Transmutation of Waste (ATW) concept which aims at destruction of key long-lived radionuclides in high-level nuclear waste (HLW), both fission products and actinides. This focus makes it different from most other transmutation concepts which concentrate primarily on actinide burning. The ATW system uses an accelerator-driven, sub-critical assembly to create an intense thermal neutron environment for radionuclide transmutation. This feature allows rapid transmutation under low-inventory system conditions, which in turn, has a direct impact on the size of chemical separations and materials handling components of the system. Inventories in ATW are factors of eight to thirty times smaller than reactor systems of equivalent thermal power. Chemical separations systems are relatively small in scale and can be optimized to achieve high decontamination factors and minimized waste streams. The low-inventory feature also directly impacts material amounts remaining in the system at its end of life. In addition to its low-inventory operation, the accelerator-driven neutron source features of ATW are key to providing a sufficient level of neutrons to allow transmutation of long-lived fission products.

  1. High level bacterial contamination of secondary school students’ mobile phones

    PubMed Central

    Kõljalg, Siiri; Mändar, Rando; Sõber, Tiina; Rööp, Tiiu; Mändar, Reet

    2017-01-01

    Introduction While contamination of mobile phones in the hospital has been found to be common in several studies, little information about bacterial abundance on phones used in the community is available. Our aim was to quantitatively determine the bacterial contamination of secondary school students’ mobile phones. Methods Altogether 27 mobile phones were studied. The contact plate method and microbial identification using MALDI-TOF mass spectrometer were used for culture studies. Quantitative PCR reaction for detection of universal 16S rRNA, Enterococcus faecalis 16S rRNA and Escherichia coli allantoin permease were performed, and the presence of tetracycline (tetA, tetB, tetM), erythromycin (ermB) and sulphonamide (sul1) resistance genes was assessed. Results We found a high median bacterial count on secondary school students’ mobile phones (10.5 CFU/cm2) and a median of 17,032 bacterial 16S rRNA gene copies per phone. Potentially pathogenic microbes (Staphylococcus aureus, Acinetobacter spp., Pseudomonas spp., Bacillus cereus and Neisseria flavescens) were found among dominant microbes more often on phones with higher percentage of E. faecalis in total bacterial 16S rRNA. No differences in contamination level or dominating bacterial species between phone owner’s gender and between phone types (touch screen/keypad) were found. No antibiotic resistance genes were detected on mobile phone surfaces. Conclusion Quantitative study methods revealed high level bacterial contamination of secondary school students’ mobile phones. PMID:28626737

  2. High-level expressing YAC vector for transgenic animal bioreactors.

    PubMed

    Fujiwara, Y; Miwa, M; Takahashi, R; Kodaira, K; Hirabayashi, M; Suzuki, T; Ueda, M

    1999-04-01

    The position effect is one major problem in the production of transgenic animals as mammary gland bioreactors. In the present study, we introduced the human growth hormone (hGH) gene into 210-kb human alpha-lactalbumin position-independent YAC vectors using homologous recombination and produced transgenic rats via microinjection of YAC DNA into rat embryos. The efficiency of producing transgenic rats with the YAC vector DNA was the same as that using plasmid constructs. All analyzed transgenic rats had one copy of the transgene and produced milk containing a high level of hGH (0.25-8.9 mg/ml). In transgenic rats with the YAC vector in which the human alpha-lactalbumin gene was replaced with the hGH gene, tissue specificity of hGH mRNA was the same as that of the endogenous rat alpha-lactalbumin gene. Thus, the 210-kb human alpha-lactalbumin YAC is a useful vector for high-level expression of foreign genes in the milk of transgenic animals.

  3. High level radioactive waste glass production and product description

    SciTech Connect

    Sproull, J.F.; Marra, S.L.; Jantzen, C.M.

    1993-12-01

    This report examines borosilicate glass as a means of immobilizing high-level radioactive wastes. Borosilicate glass will encapsulate most of the defense and some of the commercial HLW in the US. The resulting waste forms must meet the requirements of the WA-SRD and the WAPS, which include a short term PCT durability test. The waste form producer must report the composition(s) of the borosilicate waste glass(es) produced but can choose the composition(s) to meet site-specific requirements. Although the waste form composition is the primary determinant of durability, the redox state of the glass; the existence, content, and composition of crystals; and the presence of glass-in-glass phase separation can affect durability. The waste glass should be formulated to avoid phase separation regions. The ultimate result of this effort will be a waste form which is much more stable and potentially less mobile than the liquid high level radioactive waste is currently.

  4. Burning high-level TRU waste in fusion fission reactors

    NASA Astrophysics Data System (ADS)

    Shen, Yaosong

    2016-09-01

    Recently, the concept of actinide burning instead of a once-through fuel cycle for disposing spent nuclear fuel seems to get much more attention. A new method of burning high-level transuranic (TRU) waste combined with Thorium-Uranium (Th-U) fuel in the subcritical reactors driven by external fusion neutron sources is proposed in this paper. The thorium-based TRU fuel burns all of the long-lived actinides via a hard neutron spectrum while outputting power. A one-dimensional model of the reactor concept was built by means of the ONESN_BURN code with new data libraries. The numerical results included actinide radioactivity, biological hazard potential, and much higher burnup rate of high-level transuranic waste. The comparison of the fusion-fission reactor with the thermal reactor shows that the harder neutron spectrum is more efficient than the soft. The Th-U cycle produces less TRU, less radiotoxicity and fewer long-lived actinides. The Th-U cycle provides breeding of 233U with a long operation time (>20 years), hence significantly reducing the reactivity swing while improving safety and burnup.

  5. Spent fuel and high-level radioactive waste transportation report

    SciTech Connect

    Not Available

    1989-11-01

    This publication is intended to provide its readers with an introduction to the issues surrounding the subject of transportation of spent nuclear fuel and high-level radioactive waste, especially as those issues impact the southern region of the United States. It was originally issued by the Southern States Energy Board (SSEB) in July 1987 as the Spent Nuclear Fuel and High-Level Radioactive Waste Transportation Primer, a document patterned on work performed by the Western Interstate Energy Board and designed as a ``comprehensive overview of the issues.`` This work differs from that earlier effort in that it is designed for the educated layman with little or no background in nuclear waste issues. In addition, this document is not a comprehensive examination of nuclear waste issues but should instead serve as a general introduction to the subject. Owing to changes in the nuclear waste management system, program activities by the US Department of Energy and other federal agencies and developing technologies, much of this information is dated quickly. While this report uses the most recent data available, readers should keep in mind that some of the material is subject to rapid change. SSEB plans periodic updates in the future to account for changes in the program. Replacement pages sew be supplied to all parties in receipt of this publication provided they remain on the SSEB mailing list.

  6. How to achieve high-level expression of microbial enzymes

    PubMed Central

    Liu, Long; Yang, Haiquan; Shin, Hyun-dong; Chen, Rachel R.; Li, Jianghua; Du, Guocheng; Chen, Jian

    2013-01-01

    Microbial enzymes have been used in a large number of fields, such as chemical, agricultural and biopharmaceutical industries. The enzyme production rate and yield are the main factors to consider when choosing the appropriate expression system for the production of recombinant proteins. Recombinant enzymes have been expressed in bacteria (e.g., Escherichia coli, Bacillus and lactic acid bacteria), filamentous fungi (e.g., Aspergillus) and yeasts (e.g., Pichia pastoris). The favorable and very advantageous characteristics of these species have resulted in an increasing number of biotechnological applications. Bacterial hosts (e.g., E. coli) can be used to quickly and easily overexpress recombinant enzymes; however, bacterial systems cannot express very large proteins and proteins that require post-translational modifications. The main bacterial expression hosts, with the exception of lactic acid bacteria and filamentous fungi, can produce several toxins which are not compatible with the expression of recombinant enzymes in food and drugs. However, due to the multiplicity of the physiological impacts arising from high-level expression of genes encoding the enzymes and expression hosts, the goal of overproduction can hardly be achieved, and therefore, the yield of recombinant enzymes is limited. In this review, the recent strategies used for the high-level expression of microbial enzymes in the hosts mentioned above are summarized and the prospects are also discussed. We hope this review will contribute to the development of the enzyme-related research field. PMID:23686280

  7. Spent fuel and high-level radioactive waste transportation report

    SciTech Connect

    Not Available

    1990-11-01

    This publication is intended to provide its readers with an introduction to the issues surrounding the subject of transportation of spent nuclear fuel and high-level radioactive waste, especially as those issues impact the southern region of the United States. It was originally issued by the Southern States Energy Board (SSEB) in July 1987 as the Spent Nuclear Fuel and High-Level Radioactive Waste Transportation Primer, a document patterned on work performed by the Western Interstate Energy Board and designed as a ``comprehensive overview of the issues.`` This work differs from that earlier effort in that it is designed for the educated layman with little or no background in nuclear waste issues. In addition, this document is not a comprehensive examination of nuclear waste issues but should instead serve as a general introduction to the subject. Owing to changes in the nuclear waste management system, program activities by the US Department of Energy and other federal agencies and developing technologies, much of this information is dated quickly. While this report uses the most recent data available, readers should keep in mind that some of the material is subject to rapid change. SSEB plans periodic updates in the future to account for changes in the program. Replacement pages will be supplied to all parties in receipt of this publication provided they remain on the SSEB mailing list.

  8. Spent Fuel and High-Level Radioactive Waste Transportation Report

    SciTech Connect

    Not Available

    1992-03-01

    This publication is intended to provide its readers with an introduction to the issues surrounding the subject of transportation of spent nuclear fuel and high-level radioactive waste, especially as those issues impact the southern region of the United States. It was originally issued by SSEB in July 1987 as the Spent Nuclear Fuel and High-Level Radioactive Waste Transportation Primer, a document patterned on work performed by the Western Interstate Energy Board and designed as a ``comprehensive overview of the issues.`` This work differs from that earlier effort in that it is designed for the educated layman with little or no background in nuclear waste Issues. In addition. this document is not a comprehensive examination of nuclear waste issues but should instead serve as a general introduction to the subject. Owing to changes in the nuclear waste management system, program activities by the US Department of Energy and other federal agencies and developing technologies, much of this information is dated quickly. While this report uses the most recent data available, readers should keep in mind that some of the material is subject to rapid change. SSEB plans periodic updates in the future to account for changes in the program. Replacement pages will be supplied to all parties in receipt of this publication provided they remain on the SSEB mailing list.

  9. VITRIFICATION OF HIGH LEVEL WASTE AT THE SAVANNAH RIVER SITE

    SciTech Connect

    Fox, K.; Peeler, D.

    2009-06-17

    The objective of this study was to experimentally measure the properties and performance of a series of glasses with compositions that could represent high level waste Sludge Batch 5 (SB5) as vitrified at the Savannah River Site Defense Waste Processing Facility. These data were used to guide frit optimization efforts as the SB5 composition was finalized. Glass compositions for this study were developed by combining a series of SB5 composition projections with a group of candidate frits. The study glasses were fabricated using depleted uranium and their chemical compositions, crystalline contents and chemical durabilities were characterized. Trevorite was the only crystalline phase that was identified in a few of the study glasses after slow cooling, and is not of concern as spinels have been shown to have little impact on the durability of high level waste glasses. Chemical durability was quantified using the Product Consistency Test (PCT). All of the glasses had very acceptable durability performance. The results of this study indicate that a frit composition can be identified that will provide a processable and durable glass when combined with SB5.

  10. Unexpectedly high mercury level in pelleted commercial fish feed

    SciTech Connect

    Choi, M.H.; Cech, J.J. Jr.

    1998-10-01

    An unexpectedly high mercury (Hg) level was found in a pelleted commercial fish feed used to feed fish in laboratory and fish farm settings. Mean total Hg (T-Hg) concentration in the commercial fish pellets was 66 ppb. Mean total selenium (T-Se) concentration in the pellets was 1,120 ppb (ranging from 790 to 1,360 ppb). Total Hg and Se in the whole blood of Sacramento blackfish and in the fish feed were determined by inductively coupled plasma-mass spectrometry (ICP-MS). During a 10-week sampling period T-Hg in blood fluctuated between 35 and 56 ppb. A highly significant, positive correlation was found between T-Hg in the fish blood and in the fish feed through the sampling period. On the other hand, no correlation was found between T-Se in the fish feed and T-Hg or T-Se blood level. Researchers working with fish in Hg studies need to know that fish pellets may contain Hg and to consider the influence of these pellets in their results.

  11. Status of high-level waste processing at West Valley

    SciTech Connect

    Howell, A.J.; Baker, M.N. )

    1991-11-01

    The US Department of Energy is charged with the solidification of high-level liquid waste remaining from nuclear fuel reprocessing activities that were conducted at West Valley, New York, between 1966 and 1972. The 2.27 million liters (600,000 gal) of waste in an underground storage tank has separated into a sludge layer, {approximately}10% of the original volume, and a liquid layer. Prior to the high-level waste (HLW) vitrification, volume reduction of the waste is necessary. Sine May 1988, West Valley has successfully processed >1.59 million liters (420,000 gal) of HLW. Processing to date has involved the removal of {sup 139}Cs from the HLW effluent by ion exchange, evaporation to concentrate the effluent to a predetermined salt concentration, and finally cementation. This process has removed {approximately}80% of the {sup 137}Cs from the HLW liquid phase. Modifications are currently being made to begin the second phase of the HLW processing at West Valley. The second phase of HLW processing will include the removal of plutonium as well as cesium from the HLW sludge. This paper describes the progress made to date and the modifications being made to the process and to the feed stream to begin the second phase of HLW processing.

  12. Confidence Level and Sensitivity Limits in High Contrast Imaging

    SciTech Connect

    Marois, C

    2007-11-07

    In long adaptive optics corrected exposures, exoplanet detections are currently limited by speckle noise originating from the telescope and instrument optics, and it is expected that such noise will also limit future high-contrast imaging instruments for both ground and space-based telescopes. Previous theoretical analysis have shown that the time intensity variations of a single speckle follows a modified Rician. It is first demonstrated here that for a circular pupil this temporal intensity distribution also represents the speckle spatial intensity distribution at a fix separation from the point spread function center; this fact is demonstrated using numerical simulations for coronagraphic and non-coronagraphic data. The real statistical distribution of the noise needs to be taken into account explicitly when selecting a detection threshold appropriate for some desired confidence level. In this paper, a technique is described to obtain the pixel intensity distribution of an image and its corresponding confidence level as a function of the detection threshold. Using numerical simulations, it is shown that in the presence of speckles noise, a detection threshold up to three times higher is required to obtain a confidence level equivalent to that at 5{sigma} for Gaussian noise. The technique is then tested using TRIDENT CFHT and angular differential imaging NIRI Gemini adaptive optics data. It is found that the angular differential imaging technique produces quasi-Gaussian residuals, a remarkable result compared to classical adaptive optic imaging. A power-law is finally derived to predict the 1-3 x 10{sup -7} confidence level detection threshold when averaging a partially correlated non-Gaussian noise.

  13. Confidence Level and Sensitivity Limits in High Contrast Imaging

    SciTech Connect

    Marois, C; LaFreniere, D; Macintosh, B; Doyon, R

    2008-06-02

    In long adaptive optics corrected exposures, exoplanet detections are currently limited by speckle noise originating from the telescope and instrument optics, and it is expected that such noise will also limit future high-contrast imaging instruments for both ground and space-based telescopes. Previous theoretical analysis have shown that the time intensity variations of a single speckle follows a modified Rician. It is first demonstrated here that for a circular pupil this temporal intensity distribution also represents the speckle spatial intensity distribution at a fix separation from the point spread function center; this fact is demonstrated using numerical simulations for coronagraphic and non-coronagraphic data. The real statistical distribution of the noise needs to be taken into account explicitly when selecting a detection threshold appropriate for some desired confidence level. In this paper, a technique is described to obtain the pixel intensity distribution of an image and its corresponding confidence level as a function of the detection threshold. Using numerical simulations, it is shown that in the presence of speckles noise, a detection threshold up to three times higher is required to obtain a confidence level equivalent to that at 5{sigma} for Gaussian noise. The technique is then tested using TRIDENT CFHT and angular differential imaging NIRI Gemini adaptive optics data. It is found that the angular differential imaging technique produces quasi-Gaussian residuals, a remarkable result compared to classical adaptive optic imaging. A power-law is finally derived to predict the 1-3 x 10{sup -7} confidence level detection threshold when averaging a partially correlated non-Gaussian noise.

  14. Engineering neural systems for high-level problem solving.

    PubMed

    Sylvester, Jared; Reggia, James

    2016-07-01

    There is a long-standing, sometimes contentious debate in AI concerning the relative merits of a symbolic, top-down approach vs. a neural, bottom-up approach to engineering intelligent machine behaviors. While neurocomputational methods excel at lower-level cognitive tasks (incremental learning for pattern classification, low-level sensorimotor control, fault tolerance and processing of noisy data, etc.), they are largely non-competitive with top-down symbolic methods for tasks involving high-level cognitive problem solving (goal-directed reasoning, metacognition, planning, etc.). Here we take a step towards addressing this limitation by developing a purely neural framework named galis. Our goal in this work is to integrate top-down (non-symbolic) control of a neural network system with more traditional bottom-up neural computations. galis is based on attractor networks that can be "programmed" with temporal sequences of hand-crafted instructions that control problem solving by gating the activity retention of, communication between, and learning done by other neural networks. We demonstrate the effectiveness of this approach by showing that it can be applied successfully to solve sequential card matching problems, using both human performance and a top-down symbolic algorithm as experimental controls. Solving this kind of problem makes use of top-down attention control and the binding together of visual features in ways that are easy for symbolic AI systems but not for neural networks to achieve. Our model can not only be instructed on how to solve card matching problems successfully, but its performance also qualitatively (and sometimes quantitatively) matches the performance of both human subjects that we had perform the same task and the top-down symbolic algorithm that we used as an experimental control. We conclude that the core principles underlying the galis framework provide a promising approach to engineering purely neurocomputational systems for problem

  15. Department of Energy pretreatment of high-level and low-level wastes

    SciTech Connect

    McGinnis, C.P.; Hunt, R.D.

    1995-12-31

    The remediation of the 1 {times} 10{sup 8} gal of highly radioactive waste in the underground storage tanks (USTs) at five US Department of Energy (DOE) sites is one of DOE`s greatest challenges. Therefore, the DOE Office of Environmental Management has created the Tank Focus Area (TFA) to manage an integrated technology development program that results in the safe and efficient remediation of UST waste. The TFA has divided its efforts into five areas, which are safety, characterization, retrieval/closure, pretreatment, and immobilization. All DOE pretreatment activities are integrated by the Pretreatment Technical Integration Manager of the TFA. For FY 1996, the 14 pretreatment tasks are divided into 3 systems: supernate separations, sludge treatment, and solid/liquid separation. The plans and recent results of these TFA tasks, which include two 25,000-gal demonstrations and two former TFA tasks on Cs removal, are presented. The pretreatment goals are to minimize the volume of high-level waste and the radioactivity in low-level waste.

  16. Do Highly Effective Principals Also Have High Levels of Cultural Intelligence?

    ERIC Educational Resources Information Center

    Naughton, Whitney Michelle

    2010-01-01

    Purpose: The purpose of this study was to determine if elementary school principals who exhibit characteristics of highly effective principals also possess high levels of cultural intelligence. Methodology: Three instruments were used in this study, combining both qualitative and quantitative approaches to the collection of data. The first…

  17. Interleukin-1- and Type I Interferon-Dependent Enhanced Immunogenicity of an NYVAC-HIV-1 Env-Gag-Pol-Nef Vaccine Vector with Dual Deletions of Type I and Type II Interferon-Binding Proteins

    PubMed Central

    Delaloye, Julie; Filali-Mouhim, Abdelali; Cameron, Mark J.; Haddad, Elias K.; Harari, Alexandre; Goulet, Jean-Pierre; Gomez, Carmen E.; Perdiguero, Beatriz; Esteban, Mariano; Pantaleo, Giuseppe; Roger, Thierry; Sékaly, Rafick-Pierre

    2015-01-01

    ABSTRACT NYVAC, a highly attenuated, replication-restricted poxvirus, is a safe and immunogenic vaccine vector. Deletion of immune evasion genes from the poxvirus genome is an attractive strategy for improving the immunogenic properties of poxviruses. Using systems biology approaches, we describe herein the enhanced immunological profile of NYVAC vectors expressing the HIV-1 clade C env, gag, pol, and nef genes (NYVAC-C) with single or double deletions of genes encoding type I (ΔB19R) or type II (ΔB8R) interferon (IFN)-binding proteins. Transcriptomic analyses of human monocytes infected with NYVAC-C, NYVAC-C with the B19R deletion (NYVAC-C-ΔB19R), or NYVAC-C with B8R and B19R deletions (NYVAC-C-ΔB8RB19R) revealed a concerted upregulation of innate immune pathways (IFN-stimulated genes [ISGs]) of increasing magnitude with NYVAC-C-ΔB19R and NYVAC-C-ΔB8RB19R than with NYVAC-C. Deletion of B8R and B19R resulted in an enhanced activation of IRF3, IRF7, and STAT1 and the robust production of type I IFNs and of ISGs, whose expression was inhibited by anti-type I IFN antibodies. Interestingly, NYVAC-C-ΔB8RB19R induced the production of much higher levels of proinflammatory cytokines (tumor necrosis factor [TNF], interleukin-6 [IL-6], and IL-8) than NYVAC-C or NYVAC-C-ΔB19R as well as a strong inflammasome response (caspase-1 and IL-1β) in infected monocytes. Top network analyses showed that this broad response mediated by the deletion of B8R and B19R was organized around two upregulated gene expression nodes (TNF and IRF7). Consistent with these findings, monocytes infected with NYVAC-C-ΔB8RB19R induced a stronger type I IFN-dependent and IL-1-dependent allogeneic CD4+ T cell response than monocytes infected with NYVAC-C or NYVAC-C-ΔB19R. Dual deletion of type I and type II IFN immune evasion genes in NYVAC markedly enhanced its immunogenic properties via its induction of the increased expression of type I IFNs and IL-1β and make it an attractive candidate HIV

  18. Interleukin-1- and type I interferon-dependent enhanced immunogenicity of an NYVAC-HIV-1 Env-Gag-Pol-Nef vaccine vector with dual deletions of type I and type II interferon-binding proteins.

    PubMed

    Delaloye, Julie; Filali-Mouhim, Abdelali; Cameron, Mark J; Haddad, Elias K; Harari, Alexandre; Goulet, Jean-Pierre; Gomez, Carmen E; Perdiguero, Beatriz; Esteban, Mariano; Pantaleo, Giuseppe; Roger, Thierry; Sékaly, Rafick-Pierre; Calandra, Thierry

    2015-04-01

    NYVAC, a highly attenuated, replication-restricted poxvirus, is a safe and immunogenic vaccine vector. Deletion of immune evasion genes from the poxvirus genome is an attractive strategy for improving the immunogenic properties of poxviruses. Using systems biology approaches, we describe herein the enhanced immunological profile of NYVAC vectors expressing the HIV-1 clade C env, gag, pol, and nef genes (NYVAC-C) with single or double deletions of genes encoding type I (ΔB19R) or type II (ΔB8R) interferon (IFN)-binding proteins. Transcriptomic analyses of human monocytes infected with NYVAC-C, NYVAC-C with the B19R deletion (NYVAC-C-ΔB19R), or NYVAC-C with B8R and B19R deletions (NYVAC-C-ΔB8RB19R) revealed a concerted upregulation of innate immune pathways (IFN-stimulated genes [ISGs]) of increasing magnitude with NYVAC-C-ΔB19R and NYVAC-C-ΔB8RB19R than with NYVAC-C. Deletion of B8R and B19R resulted in an enhanced activation of IRF3, IRF7, and STAT1 and the robust production of type I IFNs and of ISGs, whose expression was inhibited by anti-type I IFN antibodies. Interestingly, NYVAC-C-ΔB8RB19R induced the production of much higher levels of proinflammatory cytokines (tumor necrosis factor [TNF], interleukin-6 [IL-6], and IL-8) than NYVAC-C or NYVAC-C-ΔB19R as well as a strong inflammasome response (caspase-1 and IL-1β) in infected monocytes. Top network analyses showed that this broad response mediated by the deletion of B8R and B19R was organized around two upregulated gene expression nodes (TNF and IRF7). Consistent with these findings, monocytes infected with NYVAC-C-ΔB8RB19R induced a stronger type I IFN-dependent and IL-1-dependent allogeneic CD4(+) T cell response than monocytes infected with NYVAC-C or NYVAC-C-ΔB19R. Dual deletion of type I and type II IFN immune evasion genes in NYVAC markedly enhanced its immunogenic properties via its induction of the increased expression of type I IFNs and IL-1β and make it an attractive candidate HIV

  19. High-Voltage-Input Level Translator Using Standard CMOS

    NASA Technical Reports Server (NTRS)

    Yager, Jeremy A.; Mojarradi, Mohammad M.; Vo, Tuan A.; Blalock, Benjamin J.

    2011-01-01

    proposed integrated circuit would translate (1) a pair of input signals having a low differential potential and a possibly high common-mode potential into (2) a pair of output signals having the same low differential potential and a low common-mode potential. As used here, "low" and "high" refer to potentials that are, respectively, below or above the nominal supply potential (3.3 V) at which standard complementary metal oxide/semiconductor (CMOS) integrated circuits are designed to operate. The input common-mode potential could lie between 0 and 10 V; the output common-mode potential would be 2 V. This translation would make it possible to process the pair of signals by use of standard 3.3-V CMOS analog and/or mixed-signal (analog and digital) circuitry on the same integrated-circuit chip. A schematic of the circuit is shown in the figure. Standard 3.3-V CMOS circuitry cannot withstand input potentials greater than about 4 V. However, there are many applications that involve low-differential-potential, high-common-mode-potential input signal pairs and in which standard 3.3-V CMOS circuitry, which is relatively inexpensive, would be the most appropriate circuitry for performing other functions on the integrated-circuit chip that handles the high-potential input signals. Thus, there is a need to combine high-voltage input circuitry with standard low-voltage CMOS circuitry on the same integrated-circuit chip. The proposed circuit would satisfy this need. In the proposed circuit, the input signals would be coupled into both a level-shifting pair and a common-mode-sensing pair of CMOS transistors. The output of the level-shifting pair would be fed as input to a differential pair of transistors. The resulting differential current output would pass through six standoff transistors to be mirrored into an output branch by four heterojunction bipolar transistors. The mirrored differential current would be converted back to potential by a pair of diode-connected transistors

  20. High-level fluorescence labeling of gram-positive pathogens.

    PubMed

    Aymanns, Simone; Mauerer, Stefanie; van Zandbergen, Ger; Wolz, Christiane; Spellerberg, Barbara

    2011-01-01

    Fluorescence labeling of bacterial pathogens has a broad range of interesting applications including the observation of living bacteria within host cells. We constructed a novel vector based on the E. coli streptococcal shuttle plasmid pAT28 that can propagate in numerous bacterial species from different genera. The plasmid harbors a promoterless copy of the green fluorescent variant gene egfp under the control of the CAMP-factor gene (cfb) promoter of Streptococcus agalactiae and was designated pBSU101. Upon transfer of the plasmid into streptococci, the bacteria show a distinct and easily detectable fluorescence using a standard fluorescence microscope and quantification by FACS-analysis demonstrated values that were 10-50 times increased over the respective controls. To assess the suitability of the construct for high efficiency fluorescence labeling in different gram-positive pathogens, numerous species were transformed. We successfully labeled Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus dysgalactiae subsp. equisimilis, Enterococcus faecalis, Enterococcus faecium, Streptococcus mutans, Streptococcus anginosus and Staphylococcus aureus strains utilizing the EGFP reporter plasmid pBSU101. In all of these species the presence of the cfb promoter construct resulted in high-level EGFP expression that could be further increased by growing the streptococcal and enterococcal cultures under high oxygen conditions through continuous aeration.

  1. Analysis of metagenomic data containing high biodiversity levels.

    PubMed

    Valverde, José R; Mellado, Rafael P

    2013-01-01

    In this paper we have addressed the problem of analysing Next Generation Sequencing samples with an expected large biodiversity content. We analysed several well-known 16S rRNA datasets from experimental samples, including both large and short sequences, in numbers of tens of thousands, in addition to carefully crafted synthetic datasets containing more than 7000 OTUs. From this data analysis several patterns were identified and used to develop new guidelines for experimentation in conditions of high biodiversity. We analysed the suitability of different clustering packages for these type of situations, the problem of even sampling, the relative effectiveness of Chao1 and ACE estimators as well as their effect on sampling size for a variety of population distributions. As regards practical analysis procedures, we advocated an approach that retains as much high-quality experimental data as possible. By carefully applying selection rules combining the taxonomic assignment with clustering strategies, we derived a set of recommendations for ultra-sequencing data analysis at high biodiversity levels.

  2. Exceptionally high levels of recombination across the honey bee genome.

    PubMed

    Beye, Martin; Gattermeier, Irene; Hasselmann, Martin; Gempe, Tanja; Schioett, Morten; Baines, John F; Schlipalius, David; Mougel, Florence; Emore, Christine; Rueppell, Olav; Sirviö, Anu; Guzmán-Novoa, Ernesto; Hunt, Greg; Solignac, Michel; Page, Robert E

    2006-11-01

    The first draft of the honey bee genome sequence and improved genetic maps are utilized to analyze a genome displaying 10 times higher levels of recombination (19 cM/Mb) than previously analyzed genomes of higher eukaryotes. The exceptionally high recombination rate is distributed genome-wide, but varies by two orders of magnitude. Analysis of chromosome, sequence, and gene parameters with respect to recombination showed that local recombination rate is associated with distance to the telomere, GC content, and the number of simple repeats as described for low-recombining genomes. Recombination rate does not decrease with chromosome size. On average 5.7 recombination events per chromosome pair per meiosis are found in the honey bee genome. This contrasts with a wide range of taxa that have a uniform recombination frequency of about 1.6 per chromosome pair. The excess of recombination activity does not support a mechanistic role of recombination in stabilizing pairs of homologous chromosome during chromosome pairing. Recombination rate is associated with gene size, suggesting that introns are larger in regions of low recombination and may improve the efficacy of selection in these regions. Very few transposons and no retrotransposons are present in the high-recombining genome. We propose evolutionary explanations for the exceptionally high genome-wide recombination rate.

  3. High Level Waste System Impacts from Acid Dissolution of Sludge

    SciTech Connect

    KETUSKY, EDWARD

    2006-04-20

    This research evaluates the ability of OLI{copyright} equilibrium based software to forecast Savannah River Site High Level Waste system impacts from oxalic acid dissolution of Tank 1-15 sludge heels. Without further laboratory and field testing, only the use of oxalic acid can be considered plausible to support sludge heel dissolution on multiple tanks. Using OLI{copyright} and available test results, a dissolution model is constructed and validated. Material and energy balances, coupled with the model, identify potential safety concerns. Overpressurization and overheating are shown to be unlikely. Corrosion induced hydrogen could, however, overwhelm the tank ventilation. While pH adjustment can restore the minimal hydrogen generation, resultant precipitates will notably increase the sludge volume. OLI{copyright} is used to develop a flowsheet such that additional sludge vitrification canisters and other negative system impacts are minimized. Sensitivity analyses are used to assess the processability impacts from variations in the sludge/quantities of acids.

  4. High-level iodination of monoclonal antibody fragments for radiotherapy

    SciTech Connect

    Ferens, J.M.; Krohn, K.A.; Beaumier, P.L.; Brown, J.P.; Hellstroem, I.; Hellstroem, K.E.; Carrasquillo, J.A.; Larson, S.M.

    1984-03-01

    Two different murine monoclonal antibody Fab fragments specific for p97, a melanoma-associated antigen, were labeled with I-131 at high activity levels without excessive chemical damage. Up to 20 mg of Fab were labeled with up to 300 mCi of I-131 using the chloramine-T method and large working volumes at room temperature. As much as 90% of the initial activity was recovered as labeled product. The labeled Fabs varied in their sensitivity to radioiodination damage, as measured by an in vitro cell-binding assay. Radioiodination was performed safely using a remote iodination apparatus. The final product was of radiopharmaceutical quality suitable for clinical diagnosis and experimental radiotherapy in humans.

  5. Socioeconomic studies of high-level nuclear waste disposal.

    PubMed Central

    White, G F; Bronzini, M S; Colglazier, E W; Dohrenwend, B; Erikson, K; Hansen, R; Kneese, A V; Moore, R; Page, E B; Rappaport, R A

    1994-01-01

    The socioeconomic investigations of possible impacts of the proposed repository for high-level nuclear waste at Yucca Mountain, Nevada, have been unprecedented in several respects. They bear on the public decision that sooner or later will be made as to where and how to dispose permanently of the waste presently at military weapons installations and that continues to accumulate at nuclear power stations. No final decision has yet been made. There is no clear precedent from other countries. The organization of state and federal studies is unique. The state studies involve more disciplines than any previous efforts. They have been carried out in parallel to federal studies and have pioneered in defining some problems and appropriate research methods. A recent annotated bibliography provides interested scientists with a compact guide to the 178 published reports, as well as to relevant journal articles and related documents. PMID:7971963

  6. Sandia National Laboratories' new high level acoustic test facility

    SciTech Connect

    Rogers, J. D.; Hendrick, D. M.

    1989-01-01

    A high intensity acoustic test facility has been designed and is under construction at Sandia National Laboratories in Albuquerque, NM. The chamber is designed to provide an acoustic environment of 154dB (re 20 {mu}Pa) overall sound pressure level over the bandwidth of 50 Hz to 10,000 Hz. The chamber has a volume of 16,000 cubic feet with interior dimensions of 21.6 ft {times} 24.6 ft {times} 30 ft. The construction of the chamber should be complete by the summer of 1990. This paper discusses the design goals and constraints of the facility. The construction characteristics are discussed in detail, as are the acoustic performance design characteristics. The authors hope that this work will help others in designing acoustic chambers. 12 refs., 6 figs.

  7. High level security policies for Healthcare Information Systems.

    PubMed

    Katsikas, Sokratis; Kokolakis, Spyros

    2003-01-01

    Healthcare Establishments (HCE) have are today highly dependent upon Information and Communications Technologies (ICT). This increasing reliance upon ICT has stressed the need to foster security in Healthcare Information Systems (HIS). Security policies may have a significant contribution to this effort, but they could become the cause of portability and interoperability problems. Moreover, policies that fail to take into account all the aspects of HIS security, the legal and regulatory requirements, and the existence of several stakeholders may lead to ineffective or inefficient security measures. Policies of a special category, named Generic Security Policies (GSPs), should be developed to provide policy-level harmonisation and guidance to policy-makers within HCEs. Five such policies are comparatively reviewed herein.

  8. National high-level waste systems analysis plan

    SciTech Connect

    Kristofferson, K.; Oholleran, T.P.; Powell, R.H.; Thiel, E.C.

    1995-05-01

    This document details the development of modeling capabilities that can provide a system-wide view of all US Department of Energy (DOE) high-level waste (HLW) treatment and storage systems. This model can assess the impact of budget constraints on storage and treatment system schedules and throughput. These impacts can then be assessed against existing and pending milestones to determine the impact to the overall HLW system. A nation-wide view of waste treatment availability will help project the time required to prepare HLW for disposal. The impacts of the availability of various treatment systems and throughput can be compared to repository readiness to determine the prudent application of resources or the need to renegotiate milestones.

  9. High-level theoretical rovibrational spectroscopy of HCS+ isotopologues

    NASA Astrophysics Data System (ADS)

    Schröder, B.; Sebald, P.

    2016-12-01

    In this work the rovibrational spectrum of the HCS+ molecular cation is revisited through high-level electronic structure and variational rovibrational calculations. A local potential energy function is built from explicitly correlated coupled-cluster results, incorporating corrections for core-valence, scalar relativistic and higher-order excitation effects. The computed spectroscopic parameters, based on variational calculations with Watson's isomorphic Hamiltonian for linear molecules lead to a nearly perfect agreement with experimentally reported values (Rosenbaum et al., 1989). Furthermore, the documented Fermi resonance within the (0,00, 1) / (0,20, 0) and (1,00, 1) / (1,20, 0) pairs of states is clarified. Based on a newly developed electric dipole moment function transition dipole moments of fundamental transitions are predicted for the most important isotopologues.

  10. High-level waste tank farm set point document

    SciTech Connect

    Anthony, J.A. III

    1995-01-15

    Setpoints for nuclear safety-related instrumentation are required for actions determined by the design authorization basis. Minimum requirements need to be established for assuring that setpoints are established and held within specified limits. This document establishes the controlling methodology for changing setpoints of all classifications. The instrumentation under consideration involve the transfer, storage, and volume reduction of radioactive liquid waste in the F- and H-Area High-Level Radioactive Waste Tank Farms. The setpoint document will encompass the PROCESS AREA listed in the Safety Analysis Report (SAR) (DPSTSA-200-10 Sup 18) which includes the diversion box HDB-8 facility. In addition to the PROCESS AREAS listed in the SAR, Building 299-H and the Effluent Transfer Facility (ETF) are also included in the scope.

  11. High-level tetracycline resistant Neisseria gonorrhoeae isolated in Portugal.

    PubMed

    Ferreira, E; Louro, D; Gomes, J P; Catry, M A; Pato, M V

    1997-05-01

    The first high-level tetracycline resistance (MIC > or = 16 mg/l) isolates of Neisseria gonorrhoeae (TRNG) were reported in 1990 from patients attending a Sexual Transmitted Disease (STD) Center in Lisbon. The TRNG prevalence was 4% in 1991, 5.3% in 1992 and 10,8% in 1994, exploding to 52.2% in 1995. The tet M determinant was evaluated by PCR. The digests of PCRP using HpaII produced the restriction pattern 2 for all the strains, except one (pattern 3). 78.3% of the TRNG strains were beta-lactamase producers and the 4.5 MDa penicillinase plasmid was the dominant (83%), 90% and 93.3% of the TRNG strains belonged to the auxotype NR and to the serogroup IA, respectively. The IA-8/NR class represented 58.3% of the TRNG isolates, suggesting a clonal spreading.

  12. Respiratory physiology: adaptations to high-level exercise.

    PubMed

    McKenzie, Donald C

    2012-05-01

    Most exercise scientists would agree that the physiological determinants of peak endurance performance include the capacity to transport oxygen to the working muscle, diffusion from the muscle to the mitochondria, energy production and force generation, all influenced by signals from the central nervous system. In general, the capacity of the pulmonary system far exceeds the demands required for ventilation and gas exchange during exercise. Endurance training induces large and significant adaptations within the cardiovascular, musculoskeletal and haematological systems. However, the structural and functional properties of the lung and airways do not change in response to repetitive physical activity and, in elite athletes, the pulmonary system may become a limiting factor to exercise at sea level and altitude. As a consequence to this respiratory paradox, highly trained athletes may develop intrathoracic and extrathoracic obstruction, expiratory flow limitation, respiratory muscle fatigue and exercise-induced hypoxaemia. All of these maladaptations may influence performance.

  13. SIMULANT DEVELOPMENT FOR SAVANNAH RIVER SITE HIGH LEVEL WASTE

    SciTech Connect

    Stone, M; Russell Eibling, R; David Koopman, D; Dan Lambert, D; Paul Burket, P

    2007-09-04

    The Defense Waste Processing Facility (DWPF) at the Savannah River Site vitrifies High Level Waste (HLW) for repository internment. The process consists of three major steps: waste pretreatment, vitrification, and canister decontamination/sealing. The HLW consists of insoluble metal hydroxides (primarily iron, aluminum, magnesium, manganese, and uranium) and soluble sodium salts (carbonate, hydroxide, nitrite, nitrate, and sulfate). The HLW is processed in large batches through DWPF; DWPF has recently completed processing Sludge Batch 3 (SB3) and is currently processing Sludge Batch 4 (SB4). The composition of metal species in SB4 is shown in Table 1 as a function of the ratio of a metal to iron. Simulants remove radioactive species and renormalize the remaining species. Supernate composition is shown in Table 2.

  14. 4.5 Meter high level waste canister study

    SciTech Connect

    Calmus, R. B.

    1997-10-01

    The Tank Waste Remediation System (TWRS) Storage and Disposal Project has established the Immobilized High-Level Waste (IBLW) Storage Sub-Project to provide the capability to store Phase I and II BLW products generated by private vendors. A design/construction project, Project W-464, was established under the Sub-Project to provide the Phase I capability. Project W-464 will retrofit the Hanford Site Canister Storage Building (CSB) to accommodate the Phase I I-ILW products. Project W-464 conceptual design is currently being performed to interim store 3.0 m-long BLW stainless steel canisters with a 0.61 in diameter, DOE is considering using a 4.5 in canister of the same diameter to reduce permanent disposal costs. This study was performed to assess the impact of replacing the 3.0 in canister with the 4.5 in canister. The summary cost and schedule impacts are described.

  15. High-spin level scheme of {sup 194}Pb

    SciTech Connect

    Kutsarova, T.; Stefanova, E. A.; Minkova, A.; Lalkovski, S.; Korichi, A.; Lopez-Martens, A.; Hannachi, F.; Huebel, H.; Goergen, A.; Jansen, A.; Schoenwasser, G.; Khoo, T. L.; Herskind, B.; Bergstroem, M.; Bazzacco, D.; Podolyak, Z.

    2009-01-15

    High-spin states in {sup 194}Pb have been populated in the {sup 168}Er({sup 30}Si,4n) reaction at 142 MeV. The emitted {gamma} rays were detected by the EUROBALL III multidetector array. The level scheme was considerably extended and many previously observed {gamma}-ray transitions were reordered. Four new magnetic rotational bands were observed. The energies and spins of the bandheads of all previously observed magnetic rotational bands were corrected based on the observation of new transitions. From nine observed bands, only one could not be connected to the lower lying states. Based on comparison systematics with neighboring Pb isotopes and tilted-axis cranking model calculations previously reported, configuration assignments to the observed bands have been made.

  16. The SEISMED High Level Security Policy for Health Care.

    PubMed

    Katsikas, S K

    1996-01-01

    The proliferation of the use of automated Health Information Systems in the everyday practice of health professionals has brought a number of issues related to the security of health information to a critical point. The preservation of security of health-related information can only be achieved through a concerted approach, comprising legal, organisational, technical and educational actions. These classes of actions constitute a complete "security framework", a key aspect of which is the set of rules, laws and regulations that govern the usage of information within a Health Care Establishment. This set is commonly referred to as "Security Policy". In this paper, the SEISMED High Level Security Policy for Health Care Establishments is presented.

  17. High level security policies for health care establishments.

    PubMed

    Katsikas, Sokratis; Kokolakis, Spyros

    2004-01-01

    Health Care Establishments (HCE) are today highly dependent upon Information and Communications Technologies (ICT). This increasing reliance upon ICT has stressed the need to foster security in Healthcare Information Systems (HIS). Security policies may have a significant contribution to this effort, but they could become the cause of portability and interoperability problems. Moreover, policies that fail to take into account all the aspects of HIS security, the legal and regulatory requirements, and the existence of several stakeholders may lead to ineffective or inefficient security measures. Policies of a special category, named Generic Security Policies (GSP), should be developed to provide policy-level harmonisation and guidance to policy-makers within HCE. Six such policies are comparatively reviewed herein.

  18. Midwestern High-Level Radioactive Waste Transportation Project

    SciTech Connect

    Dantoin, T.S.

    1990-12-01

    For more than half a century, the Council of State Governments has served as a common ground for the states of the nation. The Council is a nonprofit, state-supported and -directed service organization that provides research and resources, identifies trends, supplies answers and creates a network for legislative, executive and judicial branch representatives. This List of Available Resources was prepared with the support of the US Department of Energy, Cooperative Agreement No. DE-FC02-89CH10402. However, any opinions, findings, conclusions, or recommendations expressed herein are those of the author(s) and do not necessarily reflect the views of DOE. The purpose of the agreement, and reports issued pursuant to it, is to identify and analyze regional issues pertaining to the transportation of high-level radioactive waste and to inform Midwestern state officials with respect to technical issues and regulatory concerns related to waste transportation.

  19. A High-Level Language for Rule-Based Modelling

    PubMed Central

    Pedersen, Michael; Phillips, Andrew; Plotkin, Gordon D.

    2015-01-01

    Rule-based languages such as Kappa excel in their support for handling the combinatorial complexities prevalent in many biological systems, including signalling pathways. But Kappa provides little structure for organising rules, and large models can therefore be hard to read and maintain. This paper introduces a high-level, modular extension of Kappa called LBS-κ. We demonstrate the constructs of the language through examples and three case studies: a chemotaxis switch ring, a MAPK cascade, and an insulin signalling pathway. We then provide a formal definition of LBS-κ through an abstract syntax and a translation to plain Kappa. The translation is implemented in a compiler tool which is available as a web application. We finally demonstrate how to increase the expressivity of LBS-κ through embedded scripts in a general-purpose programming language, a technique which we view as generally applicable to other domain specific languages. PMID:26043208

  20. High-Level Language Production in Parkinson's Disease: A Review

    PubMed Central

    Altmann, Lori J. P.; Troche, Michelle S.

    2011-01-01

    This paper discusses impairments of high-level, complex language production in Parkinson's disease (PD), defined as sentence and discourse production, and situates these impairments within the framework of current psycholinguistic theories of language production. The paper comprises three major sections, an overview of the effects of PD on the brain and cognition, a review of the literature on language production in PD, and a discussion of the stages of the language production process that are impaired in PD. Overall, the literature converges on a few common characteristics of language production in PD: reduced information content, impaired grammaticality, disrupted fluency, and reduced syntactic complexity. Many studies also document the strong impact of differences in cognitive ability on language production. Based on the data, PD affects all stages of language production including conceptualization and functional and positional processing. Furthermore, impairments at all stages appear to be exacerbated by impairments in cognitive abilities. PMID:21860777

  1. High level radioactive waste vitrification process equipment component testing

    SciTech Connect

    Siemens, D.H.; Heath, W.O.; Larson, D.E.; Craig, S.N.; Berger, D.N.; Goles, R.W.

    1985-04-01

    Remote operability and maintainability of vitrification equipment were assessed under shielded-cell conditions. The equipment tested will be applied to immobilize high-level and transuranic liquid waste slurries that resulted from plutonium production for defense weapons. Equipment tested included: a turntable for handling waste canisters under the melter; a removable discharge cone in the melter overflow section; a thermocouple jumper that extends into a shielded cell; remote instrument and electrical connectors; remote, mechanical, and heat transfer aspects of the melter glass overflow section; a reamer to clean out plugged nozzles in the melter top; a closed circuit camera to view the melter interior; and a device to retrieve samples of the glass product. A test was also conducted to evaluate liquid metals for use in a liquid metal sealing system.

  2. A high-level language for rule-based modelling.

    PubMed

    Pedersen, Michael; Phillips, Andrew; Plotkin, Gordon D

    2015-01-01

    Rule-based languages such as Kappa excel in their support for handling the combinatorial complexities prevalent in many biological systems, including signalling pathways. But Kappa provides little structure for organising rules, and large models can therefore be hard to read and maintain. This paper introduces a high-level, modular extension of Kappa called LBS-κ. We demonstrate the constructs of the language through examples and three case studies: a chemotaxis switch ring, a MAPK cascade, and an insulin signalling pathway. We then provide a formal definition of LBS-κ through an abstract syntax and a translation to plain Kappa. The translation is implemented in a compiler tool which is available as a web application. We finally demonstrate how to increase the expressivity of LBS-κ through embedded scripts in a general-purpose programming language, a technique which we view as generally applicable to other domain specific languages.

  3. Electrophysiological correlates of high-level perception during spatial navigation

    PubMed Central

    Weidemann, Christoph T.; Mollison, Matthew V.; Kahana, Michael J.

    2009-01-01

    We studied the electrophysiological basis o object recognition by recording scalp EEG while participants played a virtual reality taxi driver game. Participants searched for passengers and stores during virtual navigation in simulated towns. We compared oscillatory brain activity for store views that were targets or non-targets (during store search) or neutral (during passenger search). Even though store category was solely defined by task context (rather than sensory cues), frontal electrophysiological activity in low frequency bands (primarily in the theta [4–8 Hz] band) reliably distinguished between target, non-target, and neutral store views. These results implicate low frequency oscillatory brain activity in frontal regions as an important variable in the study of cognitive processes involved in object recognition, categorization, and other forms of high-level perception. PMID:19293100

  4. Reinforcement learning for high-level fuzzy Petri nets.

    PubMed

    Shen, V L

    2003-01-01

    The author has developed a reinforcement learning algorithm for the high-level fuzzy Petri net (HLFPN) models in order to perform structure and parameter learning simultaneously. In addition to the HLFPN itself, the difference and similarity among a variety of subclasses concerning Petri nets are also discussed. As compared with the fuzzy adaptive learning control network (FALCON), the HLFPN model preserves the advantages that: 1) it offers more flexible learning capability because it is able to model both IF-THEN and IF-THEN-ELSE rules; 2) it allows multiple heterogeneous outputs to be drawn if they exist; 3) it offers a more compact data structure for fuzzy production rules so as to save information storage; and 4) it is able to learn faster due to its structural reduction. Finally, main results are presented in the form of seven propositions and supported by some experiments.

  5. High-level waste program integration within the DOE complex

    SciTech Connect

    Valentine, J.H.; Davis, N.R.; Malone, K.; Schaus, P.S.

    1998-03-01

    Eleven major Department of Energy (DOE) site contractors were chartered by the Assistant Secretary to use a systems engineering approach to develop and evaluate technically defensible cost savings opportunities across the complex. Known as the complex-wide Environmental Management Integration (EMI), this process evaluated all the major DOE waste streams including high level waste (HLW). Across the DOE complex, this waste stream has the highest life cycle cost and is scheduled to take until at least 2035 before all HLW is processed for disposal. Technical contract experts from the four DOE sites that manage high level waste participated in the integration analysis: Hanford, Savannah River Site (SRS), Idaho National Engineering and Environmental Laboratory (INEEL), and West Valley Demonstration Project (WVDP). In addition, subject matter experts from the Yucca Mountain Project and the Tanks Focus Area participated in the analysis. Also, departmental representatives from the US Department of Energy Headquarters (DOE-HQ) monitored the analysis and results. Workouts were held throughout the year to develop recommendations to achieve a complex-wide integrated program. From this effort, the HLW Environmental Management (EM) Team identified a set of programmatic and technical opportunities that could result in potential cost savings and avoidance in excess of $18 billion and an accelerated completion of the HLW mission by seven years. The cost savings, schedule improvements, and volume reduction are attributed to a multifaceted HLW treatment disposal strategy which involves waste pretreatment, standardized waste matrices, risk-based retrieval, early development and deployment of a shipping system for glass canisters, and reasonable, low cost tank closure.

  6. A High School Project Seminar on Sea Level Rise

    NASA Astrophysics Data System (ADS)

    Seitz, M.; Bosch, W.

    2012-04-01

    In Bavaria the curriculum of the upper grade of high school includes a so called project seminar, running over one and a half year. The aims of the seminar are to let the pupils learn to work on a specific topic, to organize themselves in a team, to improve their soft skills and become familiar with the working life. The topic of the project seminar, jointly organized by the Bertold-Brecht-Gymnasium in Munich and the Deutsche Geodätische Forschungsinstitut (DGFI) was on the "Global sea level rise". A team of 13 pupils computed the mean sea level rise by using on the one hand altimetry data of TOPEX, Jason-1 and Jason2 and on the other hand data of globally distributed tide gauges, corrected for vertical crustal movements derived from GPS products. The results of the two independent approaches were compared with each other and discussed considering also statements and discussions found in press, TV, and the web. Finally, a presentation was prepared and presented at school.

  7. Hemipelvectomy: high-level amputation surgery and prosthetic rehabilitation.

    PubMed

    Houdek, Matthew T; Kralovec, Michael E; Andrews, Karen L

    2014-07-01

    The hemipelvectomy, most commonly performed for pelvic tumor resection, is one of the most technically demanding and invasive surgical procedures performed today. Adequate soft tissue coverage and wound complications after hemipelvectomy are important considerations. Rehabilitation after hemipelvectomy is optimally managed by a multidisciplinary integrated team. Understanding the functional outcomes for this population assists the rehabilitation team to counsel patients, plan goals, and determine discharge needs. The most important rehabilitation goal is the optimal restoration of the patient's functional independence. Factors such as age, sex, etiology, level of amputation, and general health play important roles in determining prosthetic use. The three main criteria for successful prosthetic rehabilitation of patients with high-level amputation are comfort, function, and cosmesis. Recent advances in hip and knee joints have contributed to increased function. Prosthetic use after hemipelvectomy improves balance and decreases the need for a gait aid. Using a prosthesis helps maintain muscle strength and tone, cardiovascular health, and functional mobility. With new advances in prosthetic components, patients are choosing to use their prostheses for primary mobility.

  8. Vestibular contributions to high-level sensorimotor functions.

    PubMed

    Medendorp, W Pieter; Selen, Luc J P

    2017-02-02

    The vestibular system, which detects motion and orientation of the head in space, is known to be important in controlling gaze to stabilize vision, to ensure postural stability and to provide our sense of self-motion. While the brain's computations underlying these functions are extensively studied, the role of the vestibular system in higher level sensorimotor functions is less clear. This review covers new research on the vestibular influence on perceptual judgments, motor decisions, and the ability to learn multiple motor actions. Guided by concepts such as optimization, inference, estimation and control, we focus on how the brain determines causal relationships between memorized and visual representations in the updating of visual space, and how vestibular, visual and efferent motor information are integrated in the estimation of body motion. We also discuss evidence that these computations involve multiple coordinate representations, some of which can be probed in parietal cortex using neuronal oscillations derived from EEG. In addition, we describe work on decision making during self-motion, showing a clear modulation of bottom-up acceleration signals on decisions in the saccadic system. Finally, we consider the importance of vestibular signals as contextual cues in motor learning and recall. Taken together, these results emphasize the impact of vestibular information on high-level sensorimotor functions, and identify future directions for theoretical, behavioral, and neurophysiological investigations.

  9. Interpreting methamphetamine levels in a high-use community.

    PubMed

    Chiaia-Hernandez, Aurea C; Banta-Green, Caleb J; Field, Jennifer A

    2011-11-01

    Illicit drug use is a largely hidden phenomenon, and population measures are notoriously problematic. Reliable and valid data for local, regional, and national public health and other interventions are needed. To address this information, we examined temporal trends within and across weeks in methamphetamine (MA) in a single location in order to inform a sampling plan for understanding long-term trends in MA use based on sampling raw influent to wastewater treatment plants. The measured concentrations in the wastewater are used to estimate the total mass of MA consumed rather than the number of doses due to the uncertainty surrounding methamphetamine purity, mass of MA per dose, and the number of doses used per day. Results from a region with high levels of MA use indicate that MA levels do not differ significantly between weekdays and weekends (p = 0.1), consistent with a predominately regular, daily use pattern use. The potential contribution of legal sales of D: - and L: -MA to the mass of MA consumed within the community was estimated to range from 3-8%. Limitations and uncertainties associated with estimating the mass of MA consumption include small contributions of prescription and over-the-counter drugs that are metabolized to MA as well as measurement and sampling variability.

  10. The LHCb Data Acquisition and High Level Trigger Processing Architecture

    NASA Astrophysics Data System (ADS)

    Frank, M.; Gaspar, C.; Jost, B.; Neufeld, N.

    2015-12-01

    The LHCb experiment at the LHC accelerator at CERN collects collisions of particle bunches at 40 MHz. After a first level of hardware trigger with an output rate of 1 MHz, the physically interesting collisions are selected by running dedicated trigger algorithms in the High Level Trigger (HLT) computing farm. This farm consists of up to roughly 25000 CPU cores in roughly 1750 physical nodes each equipped with up to 4 TB local storage space. This work describes the LHCb online system with an emphasis on the developments implemented during the current long shutdown (LS1). We will elaborate the architecture to treble the available CPU power of the HLT farm and the technicalities to determine and verify precise calibration and alignment constants which are fed to the HLT event selection procedure. We will describe how the constants are fed into a two stage HLT event selection facility using extensively the local disk buffering capabilities on the worker nodes. With the installed disk buffers, the CPU resources can be used during periods of up to ten days without beams. These periods in the past accounted to more than 70% of the total time.

  11. Seeking explanations for high levels of infant mortality in Pakistan.

    PubMed

    Sathar, Z A

    1987-01-01

    Data from the Fertility Module of the 1979 Population, Labour Force and Migration (PLM) Survey of Pakistan were analyzed to determine which of 4 factors were primarily responsible for the high infant mortality rate. The factors examined were poverty, childbearing and childrearing practices, distribution of health care and lack of individual attention given to children due to ignorance. These items were presented in a discussion format. Infant mortality in Pakistan is high at about 125-140/1000, for a country with mid-level per capita income. Income was not a good indicator of child mortality, primarily because it was difficult to determine, particularly in rural areas where non-cash income predominates. Wealth and status were good indicators of child survival. Child-rearing practices were somewhat important, as judged by birth order, breastfeeding duration and gender. Childbearing practices as shown by spacing were important determinants of survival. Health care facilities were somewhat important, indicated by higher mortality in rural areas. Rural neonates die from tetanus due to lack of immunization, or later from diarrheal disease due to lack of potable water or poor weaning practices. Maternal education was a strong indicator of survival, much more so than paternal education. Similarly, female heads of households increased survival, probably because they control financial allocations. The study suggested that rather than attempting to eliminate poverty overall, improvements in maternal education, nutrition, health care facilities and their use, and childbearing and child-rearing methods would do more to improve child survival in Pakistan.

  12. Ultrafilter Conditions for High Level Waste Sludge Processing

    SciTech Connect

    Geeting, John GH; Hallen, Richard T.; Peterson, Reid A.

    2006-08-28

    An evaluation of the optimal filtration conditions was performed based on test data obtained from filtration of a High Level Waste Sludge sample from the Hanford tank farms. This evaluation was performed using the anticipated configuration for the Waste Treatment Plant at the Hanford site. Testing was performed to identify the optimal pressure drop and cross flow velocity for filtration at both high and low solids loading. However, this analysis indicates that the actual filtration rate achieved is relatively insensitive to these conditions under anticipated operating conditions. The maximum filter flux was obtained by adjusting the system control valve pressure from 400 to 650 kPa while the filter feed concentration increased from 5 to 20 wt%. However, operating the system with a constant control valve pressure drop of 500 kPa resulted in a less than 1% reduction in the average filter flux. Also note that allowing the control valve pressure to swing as much as +/- 20% resulted in less than a 5% decrease in filter flux.

  13. Sea level rise Contribution from High Mountain Asia by 20150

    NASA Astrophysics Data System (ADS)

    Zhao, Liyun; Moore, John; Ding, Ran

    2015-04-01

    We estimate individual area and volume change by 2050 of all 83,460 glaciers of high mountain Asia (HMA), with a total area of 118,263 km2, delineated in the Randolph Glacier Inventory version 4.0 which separates glacier complexes in its previous version into individual glaciers. We used the 25 km resolution regional climate model RegCM 3.0 temperature and precipitation change projections forced by the IPCC A1B scenario. Glacier simulations were based on a novel surface mass balance-altitude parameterization fitted to observational data, and various volume-area scaling approaches using Shuttle Radar Topography Mission surface topography of each individual glacier. We generate mass balance-altitude relations for all the glaciers by region using nearest available glacier measurements. Two method are used to model the Equilibrium line altitude (ELA) variation. One is to use ELA sensitivities to temperature and precipitation change vary by region based on the relative importance of sublimation and melting processes. The other is solved ELA implicitly for every year using the temperature at ELA and Degree Day model. We project total glacier area loss in high mountain Asia in 2050 to be 22% of their extent in 2000, and they will contribute 5-8 mm to global sea level rise.

  14. Control of high level radioactive waste-glass melters

    SciTech Connect

    Bickford, D.F.; Choi, A.S.

    1991-01-01

    Slurry Fed Melters (SFM) are being developed in the United States, Europe and Japan for the conversion of high-level radioactive waste to borosilicate glass for permanent disposal. The high transition metal, noble metal, nitrate, organic, and sulfate contents of these wastes lead to unique melter redox control requirements. Pilot waste-glass melter operations have indicated the possibility of nickel sulfide or noble-metal fission-product accumulation on melter floors, which can lead to distortion of electric heating patterns, and decrease melter life. Sulfide formation is prevented by control of the redox chemistry of the melter feed. The redox state of waste-glass melters is determined by balance between the reducing potential of organic compounds in the feed, and the oxidizing potential of gases above the melt, and nitrates and polyvalent elements in the waste. Semiquantitative models predicting limitations of organic content have been developed based on crucible testing. Computerized thermodynamic computations are being developed to predict the sequence and products of redox reactions and is assessing process variations. Continuous melter test results have been compared to improved computer staged-thermodynamic-models of redox behavior. Feed chemistry control to prevent sulfide and moderate noble metal accumulations are discussed. 17 refs., 3 figs.

  15. High dietary fiber intake prevents stroke at a population level.

    PubMed

    Casiglia, Edoardo; Tikhonoff, Valérie; Caffi, Sandro; Boschetti, Giovanni; Grasselli, Carla; Saugo, Mario; Giordano, Nunzia; Rapisarda, Valentina; Spinella, Paolo; Palatini, Paolo

    2013-10-01

    This research was aimed at clarifying whether high dietary fiber intake has an impact on incidence and risk of stroke at a population level. In 1647 unselected subjects, dietary fiber intake (DFI) was detected in a 12-year population-based study, using other dietary variables, anagraphics, biometrics, blood pressure, heart rate, blood lipids, glucose, insulin, uricaemia, fibrinogenaemia, erytrosedimentation rate, diabetes, insulin resistance, smoking, pulmonary disease and left ventricular hypertrophy as covariables. In adjusted Cox models, high DFI reduced the risk of stroke. In analysis based on quintiles of fiber intake adjusted for confounders, HR for incidence of stroke was lower when the daily intake of soluble fiber was >25 g or that of insoluble fiber was >47 g. In multivariate analyses, using these values as cut-off of DFI, the risk of stroke was lower in those intaking more that the cut-off of soluble (HR 0.31, 0.17-0.55) or insoluble (HR 0.35, 0.19-0.63) fiber. Incidence of stroke was also lower (-50%, p < 0.003 and -46%, p < 0.01, respectively). Higher dietary DFI is inversely and independently associated to incidence and risk of stroke in general population. Copyright © 2012 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

  16. High level waste characterization in support of low level waste certification. I. HLW supernate radionuclide characterization

    SciTech Connect

    Jamison, M.E.; d`Entremont, P.D.; Clemmons, J.S.; Bess, C.E.; Brown, D.F.

    1994-07-08

    High Level Waste Programs has radioactive waste storage, treatment and processing facilities that are located in the F and H Areas at the Savannah River Site. These facilities include the Effluent Treatment Facility (ETF), F and H Area Tank Farms, Extended Sludge Processing (ESP), and In-Tank Precipitation (ITP). Job wastes are generated from operation, maintenance, and construction activities inside radiological areas. These items may have been contaminated with radioactive supernate, salt, and sludge material. Most of these wastes will be disposed of in the E-area Vaults. Therefore, an isotopic and hazardous characterization must be performed. The characterization of HLW supernate radionuclides is discussed in Chapter I. The characterization for salt and sludge phases, which can also contaminate LLW, will be included in other Chapters.

  17. Defense High Level Waste Disposal Container System Description

    SciTech Connect

    2000-10-12

    The Defense High Level Waste Disposal Container System supports the confinement and isolation of waste within the Engineered Barrier System of the Monitored Geologic Repository (MGR). Disposal containers are loaded and sealed in the surface waste handling facilities, transferred to the underground through the accesses using a rail mounted transporter, and emplaced in emplacement drifts. The defense high level waste (HLW) disposal container provides long-term confinement of the commercial HLW and defense HLW (including immobilized plutonium waste forms (IPWF)) placed within disposable canisters, and withstands the loading, transfer, emplacement, and retrieval loads and environments. U.S. Department of Energy (DOE)-owned spent nuclear fuel (SNF) in disposable canisters may also be placed in a defense HLW disposal container along with commercial HLW waste forms, which is known as 'co-disposal'. The Defense High Level Waste Disposal Container System provides containment of waste for a designated period of time, and limits radionuclide release. The disposal container/waste package maintains the waste in a designated configuration, withstands maximum handling and rockfall loads, limits the individual canister temperatures after emplacement, resists corrosion in the expected handling and repository environments, and provides containment of waste in the event of an accident. Defense HLW disposal containers for HLW disposal will hold up to five HLW canisters. Defense HLW disposal containers for co-disposal will hold up to five HLW canisters arranged in a ring and one DOE SNF canister in the ring. Defense HLW disposal containers also will hold two Multi-Canister Overpacks (MCOs) and two HLW canisters in one disposal container. The disposal container will include outer and inner cylinders, outer and inner cylinder lids, and may include a canister guide. An exterior label will provide a means by which to identify the disposal container and its contents. Different materials

  18. Review of high-level waste form properties. [146 bibliographies

    SciTech Connect

    Rusin, J.M.

    1980-12-01

    This report is a review of waste form options for the immobilization of high-level-liquid wastes from the nuclear fuel cycle. This review covers the status of international research and development on waste forms as of May 1979. Although the emphasis in this report is on waste form properties, process parameters are discussed where they may affect final waste form properties. A summary table is provided listing properties of various nuclear waste form options. It is concluded that proposed waste forms have properties falling within a relatively narrow range. In regard to crystalline versus glass waste forms, the conclusion is that either glass of crystalline materials can be shown to have some advantage when a single property is considered; however, at this date no single waste form offers optimum properties over the entire range of characteristics investigated. A long-term effort has been applied to the development of glass and calcine waste forms. Several additional waste forms have enough promise to warrant continued research and development to bring their state of development up to that of glass and calcine. Synthetic minerals, the multibarrier approach with coated particles in a metal matrix, and high pressure-high temperature ceramics offer potential advantages and need further study. Although this report discusses waste form properties, the total waste management system should be considered in the final selection of a waste form option. Canister design, canister materials, overpacks, engineered barriers, and repository characteristics, as well as the waste form, affect the overall performance of a waste management system. These parameters were not considered in this comparison.

  19. Activity profile of high-level Australian lacrosse players.

    PubMed

    Polley, Chris S; Cormack, Stuart J; Gabbett, Tim J; Polglaze, Ted

    2015-01-01

    Despite lacrosse being one of the fastest growing team sports in the world, there is a paucity of information detailing the activity profile of high-level players. Microtechnology systems (global positioning systems and accelerometers) provide the opportunity to obtain detailed information on the activity profile in lacrosse. Therefore, this study aimed to analyze the activity profile of lacrosse match-play using microtechnology. Activity profile variables assessed relative to minutes of playing time included relative distance (meter per minute), distance spent standing (0-0.1 m·min), walking (0.2-1.7 m·min), jogging (1.8-3.2 m·min), running (3.3-5.6 m·min), sprinting (≥5.7 m·min), number of high, moderate, low accelerations and decelerations, and player load (PL per minute), calculated as the square root of the sum of the squared instantaneous rate of change in acceleration in 3 vectors (medio-lateral, anterior-posterior, and vertical). Activity was recorded from 14 lacrosse players over 4 matches during a national tournament. Players were separated into positions of attack, midfield, or defense. Differences (effect size [ES] ± 90% confidence interval) between positions and periods of play were considered likely positive when there was ≥75% likelihood of the difference exceeding an ES threshold of 0.2. Midfielders had likely covered higher (mean ± SD) meters per minute (100 ± 11) compared with attackers (87 ± 14; ES = 0.89 ± 1.04) and defenders (79 ± 14; ES = 1.54 ± 0.94) and more moderate and high accelerations and decelerations. Almost all variables across positions were reduced in quarter 4 compared with quarter 1. Coaches should accommodate for positional differences when preparing lacrosse players for competition.

  20. Impact of high electromagnetic field levels on childhood leukemia incidence.

    PubMed

    Teepen, Jop C; van Dijck, Jos A A M

    2012-08-15

    The increasing exposure to electromagnetic fields (EMFs) has raised concern, as increased exposure may result in an increased risk of childhood leukemia (CL). Besides a short introduction of CL and EMF, our article gives an evaluation of the evidence of a causal relation between EMF and CL by critically appraising the epidemiological and biological evidence. The potential impact is also estimated by the population attributable risk. The etiology of CL is largely unknown, but is probably multifactorial. EMF may be one of the environmental exposures involved. Three pooled analyses of case-control studies showed a 1.4- to 1.7-fold increased CL risk for extremely low-frequency EMF (ELF-EMF) exposure levels above 0.3 μT. Several biases may have played a role in these studies, but are unlikely to fully explain the increased risk. For effects of radiofrequency ELF evidence is lacking. None of the proposed biological mechanisms by which ELF-EMF might cause CL have been confirmed. The estimated overall population attributable risk was 1.9%, with the highest estimates in Northern America and Brazil (4.2% and 4.1%, respectively). The potential impact of EMF exposure on public health is probably limited, although in some countries exposure might be relatively high and thus might have a more substantial impact. We recommend nationwide surveys to gain more insight into the contemporary exposure levels among children. Reducing exposure from power lines near densely populated areas and schools is advised. Future epidemiological studies should focus on limiting bias. Copyright © 2012 UICC.

  1. Cementitious Grout for Closing SRS High Level Waste Tanks - 12315

    SciTech Connect

    Langton, C.A.; Stefanko, D.B.; Burns, H.H.; Waymer, J.; Mhyre, W.B.; Herbert, J.E.; Jolly, J.C. Jr.

    2012-07-01

    In 1997, the first two United States Department of Energy (US DOE) high level waste tanks (Tanks 17-F and 20-F: Type IV, single shell tanks) were taken out of service (permanently closed) at the Savannah River Site (SRS). In 2012, the DOE plans to remove from service two additional Savannah River Site (SRS) Type IV high-level waste tanks, Tanks 18-F and 19-F. These tanks were constructed in the late 1950's and received low-heat waste and do not contain cooling coils. Operational closure of Tanks 18-F and 19-F is intended to be consistent with the applicable requirements of the Resource Conservation and Recovery Act (RCRA) and the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) and will be performed in accordance with South Carolina Department of Health and Environmental Control (SCDHEC). The closure will physically stabilize two 4.92E+04 cubic meter (1.3 E+06 gallon) carbon steel tanks and isolate and stabilize any residual contaminants left in the tanks. Ancillary equipment abandoned in the tanks will also be filled to the extent practical. A Performance Assessment (PA) has been developed to assess the long-term fate and transport of residual contamination in the environment resulting from the operational closure of the F-Area Tank Farm (FTF) waste tanks. Next generation flowable, zero-bleed cementitious grouts were designed, tested, and specified for closing Tanks 18-F and 19-F and for filling the abandoned equipment. Fill requirements were developed for both the tank and equipment grouts. All grout formulations were required to be alkaline with a pH of 12.4 and to be chemically reducing with a reduction potential (Eh) of -200 to -400. Grouts with this chemistry stabilize potential contaminants of concern. This was achieved by including Portland cement and Grade 100 slag in the mixes, respectively. Ingredients and proportions of cementitious reagents were selected and adjusted to support the mass placement strategy developed by

  2. Transient - Photoresistance Spectroscopy of Deep Levels in High Resistivity Semiconductors

    NASA Astrophysics Data System (ADS)

    Seabaugh, Alan Carter

    A new photoinduced transient-resistance technique is proposed and examined for use in characterizing high -resistivity, short-lifetime semiconductors. In this technique, termed photoresistance deep-level transient spectroscopy (PR-DLTS), an optical pulse is used to generate excess carriers which are trapped by deep levels in the material. The ac resistance of the specimen is monitored, and the resistance transient which occurs after the illumination ends is signal processed in the same way as the capacitance transient in DLTS. A phenomenological model of the phototransient is used to derive an expression for the decay transient. Two limiting forms for the transient are predicted depending upon whether the emitted carriers are collected at the device contacts, or recombine in the bulk before being collected. In order to test this model, several methods for distinguishing between the predicted decays of the form exp(-(lamda)t) and (lamda)exp(-(lamda)t) were developed ((lamda) is the reciprocal time constant for the decay). The measured results are consistent with the predictions of the model, but an 'ideal' sample has not been found to fully test the predictions. On Au-doped Si, excellent agreement between published emission rates for the Au-acceptor and the apparent emission rates measured by PR-DLTS is found. In addition, PR-DLTS data for the Cr-related deep level in semi-insulating GaAs is consistent with published DLTS results on conducting GaAs. Comparison of the PR-DLTS technique with the dc current-transient technique, photoinduced transient spectroscopy (PITS), shows that the two techniques are sensitive to the same trapping/detrapping phenomena. Finally, the results of a comparative study of commercial semi-insulating GaAs are reported. Nineteen specimens from ten suppliers were examined using the PR -DLTS technique, including material grown by the horizontal Bridgman (HB) and liquid-encapsulated Czochralski (LEC) technique, both with and without Cr

  3. Interventions for Individuals With High Levels of Needle Fear

    PubMed Central

    Noel, Melanie; Taddio, Anna; Antony, Martin M.; Asmundson, Gordon J.G.; Riddell, Rebecca Pillai; Chambers, Christine T.; Shah, Vibhuti

    2015-01-01

    Background: This systematic review evaluated the effectiveness of exposure-based psychological and physical interventions for the management of high levels of needle fear and/or phobia and fainting in children and adults. Design/Methods: A systematic review identified relevant randomized and quasi-randomized controlled trials of children, adults, or both with high levels of needle fear, including phobia (if not available, then populations with other specific phobias were included). Critically important outcomes were self-reported fear specific to the feared situation and stimulus (psychological interventions) or fainting (applied muscle tension). Data were pooled using standardized mean difference (SMD) or relative risk with 95% confidence intervals. Results: The systematic review included 11 trials. In vivo exposure-based therapy for children 7 years and above showed benefit on specific fear (n=234; SMD: −1.71 [95% CI: −2.72, −0.7]). In vivo exposure-based therapy with adults reduced fear of needles posttreatment (n=20; SMD: −1.09 [−2.04, −0.14]) but not at 1-year follow-up (n=20; SMD: −0.28 [−1.16, 0.6]). Compared with single session, a benefit was observed for multiple sessions of exposure-based therapy posttreatment (n=93; SMD: −0.66 [−1.08, −0.24]) but not after 1 year (n=83; SMD: −0.37 [−0.87, 0.13]). Non in vivo e.g., imaginal exposure-based therapy in children reduced specific fear posttreatment (n=41; SMD: −0.88 [−1.7, −0.05]) and at 3 months (n=24; SMD: −0.89 [−1.73, −0.04]). Non in vivo exposure-based therapy for adults showed benefit on specific fear (n=68; SMD: −0.62 [−1.11, −0.14]) but not procedural fear (n=17; SMD: 0.18 [−0.87, 1.23]). Applied tension showed benefit on fainting posttreatment (n=20; SMD: −1.16 [−2.12, −0.19]) and after 1 year (n=20; SMD: −0.97 [−1.91, −0.03]) compared with exposure alone. Conclusions: Exposure-based psychological interventions and applied muscle tension show

  4. Spent nuclear fuel project high-level information management plan

    SciTech Connect

    Main, G.C.

    1996-09-13

    This document presents the results of the Spent Nuclear Fuel Project (SNFP) Information Management Planning Project (IMPP), a short-term project that identified information management (IM) issues and opportunities within the SNFP and outlined a high-level plan to address them. This high-level plan for the SNMFP IM focuses on specific examples from within the SNFP. The plan`s recommendations can be characterized in several ways. Some recommendations address specific challenges that the SNFP faces. Others form the basis for making smooth transitions in several important IM areas. Still others identify areas where further study and planning are indicated. The team`s knowledge of developments in the IM industry and at the Hanford Site were crucial in deciding where to recommend that the SNFP act and where they should wait for Site plans to be made. Because of the fast pace of the SNFP and demands on SNFP staff, input and interaction were primarily between the IMPP team and members of the SNFP Information Management Steering Committee (IMSC). Key input to the IMPP came from a workshop where IMSC members and their delegates developed a set of draft IM principles. These principles, described in Section 2, became the foundation for the recommendations found in the transition plan outlined in Section 5. Availability of SNFP staff was limited, so project documents were used as a basis for much of the work. The team, realizing that the status of the project and the environment are continually changing, tried to keep abreast of major developments since those documents were generated. To the extent possible, the information contained in this document is current as of the end of fiscal year (FY) 1995. Programs and organizations on the Hanford Site as a whole are trying to maximize their return on IM investments. They are coordinating IM activities and trying to leverage existing capabilities. However, the SNFP cannot just rely on Sitewide activities to meet its IM requirements

  5. High resolution geodetic techniques for monitoring fluid levels over time

    NASA Astrophysics Data System (ADS)

    Hare, Jennifer Thompson

    1998-11-01

    In the first study, a novel surveillance technique is developed in which surface gravity observations are used to monitor the progress of a gas cap waterflood in the 8200 ft (2500 m) deep Prudhoe Bay reservoir, Alaska. This cost-effective method requires that high-precision gravity surveys be repeated every 3 to 5 years. Differences in the gravity field with time reflect changes in the reservoir fluid density distribution. A preliminary field test at Prudhoe Bay indicates survey accuracy of 5 to 10 mu Gal can be achieved for gravity data using a modified Lacoste & Romberg "G" type meter or Scintrex CG-3M combined with Global Positioning System (GPS) positioning. Forward gravity modeling of a suite of reservoir simulations of the proposed waterflood predicts variation in surface measurements of 100 mu Gal after 5 years of injection, and 180 to 250 mu Gal after 15 years. A constrained, least-squares method is used to invert synthetic gravity data for subsurface density distributions. The modeling procedure has been formulated to allow testing of the models for sensitivity to gravity sampling patterns, noise characteristics, and various constraints on model parameters such as density range, total mass, and model moment of inertia. Horizontal feature resolution of the waterflood is about 5000 ft (1520 m) for constrained inverse models from synthetic gravity with 5 mu Gal standard deviation noise. Results of the modeling indicate that inversion of time-lapse gravity data is a viable and promising technique for monitoring reservoir gas cap waterfloods. In the second study, the problem of how to estimate ancient lake levels from the geomorphology of remnant shoreline terraces is investigated. High resolution, GPS controlled, topographic data from around the highstand shoreline of Pleistocene Lake Lahontan in western Nevada provide the means for isolating coherent terrace features which are related to the paleoshoreline level. Determination of an unambiguous point or

  6. High-level disinfection of gastrointestinal endoscope reprocessing

    PubMed Central

    Chiu, King-Wah; Lu, Lung-Sheng; Chiou, Shue-Shian

    2015-01-01

    High level disinfection (HLD) of the gastrointestinal (GI) endoscope is not simply a slogan, but rather is a form of experimental monitoring-based medicine. By definition, GI endoscopy is a semicritical medical device. Hence, such medical devices require major quality assurance for disinfection. And because many of these items are temperature sensitive, low-temperature chemical methods, such as liquid chemical germicide, must be used rather than steam sterilization. In summarizing guidelines for infection prevention and control for GI endoscopy, there are three important steps that must be highlighted: manual washing, HLD with automated endoscope reprocessor, and drying. Strict adherence to current guidelines is required because compared to any other medical device, the GI endoscope is associated with more outbreaks linked to inadequate cleaning or disinfecting during HLD. Both experimental evaluation on the surveillance bacterial cultures and in-use clinical results have shown that, the monitoring of the stringent processes to prevent and control infection is an essential component of the broader strategy to ensure the delivery of safe endoscopy services, because endoscope reprocessing is a multistep procedure involving numerous factors that can interfere with its efficacy. Based on our years of experience in the surveillance of culture monitoring of endoscopic reprocessing, we aim in this study to carefully describe what details require attention in the GI endoscopy disinfection and to share our experience so that patients can be provided with high quality and safe medical practices. Quality management encompasses all aspects of pre- and post-procedural care including the efficiency of the endoscopy unit and reprocessing area, as well as the endoscopic procedure itself. PMID:25699232

  7. High-level disinfection of gastrointestinal endoscope reprocessing.

    PubMed

    Chiu, King-Wah; Lu, Lung-Sheng; Chiou, Shue-Shian

    2015-02-20

    High level disinfection (HLD) of the gastrointestinal (GI) endoscope is not simply a slogan, but rather is a form of experimental monitoring-based medicine. By definition, GI endoscopy is a semicritical medical device. Hence, such medical devices require major quality assurance for disinfection. And because many of these items are temperature sensitive, low-temperature chemical methods, such as liquid chemical germicide, must be used rather than steam sterilization. In summarizing guidelines for infection prevention and control for GI endoscopy, there are three important steps that must be highlighted: manual washing, HLD with automated endoscope reprocessor, and drying. Strict adherence to current guidelines is required because compared to any other medical device, the GI endoscope is associated with more outbreaks linked to inadequate cleaning or disinfecting during HLD. Both experimental evaluation on the surveillance bacterial cultures and in-use clinical results have shown that, the monitoring of the stringent processes to prevent and control infection is an essential component of the broader strategy to ensure the delivery of safe endoscopy services, because endoscope reprocessing is a multistep procedure involving numerous factors that can interfere with its efficacy. Based on our years of experience in the surveillance of culture monitoring of endoscopic reprocessing, we aim in this study to carefully describe what details require attention in the GI endoscopy disinfection and to share our experience so that patients can be provided with high quality and safe medical practices. Quality management encompasses all aspects of pre- and post-procedural care including the efficiency of the endoscopy unit and reprocessing area, as well as the endoscopic procedure itself.

  8. Deep borehole disposal of high-level radioactive waste.

    SciTech Connect

    Stein, Joshua S.; Freeze, Geoffrey A.; Brady, Patrick Vane; Swift, Peter N.; Rechard, Robert Paul; Arnold, Bill Walter; Kanney, Joseph F.; Bauer, Stephen J.

    2009-07-01

    Preliminary evaluation of deep borehole disposal of high-level radioactive waste and spent nuclear fuel indicates the potential for excellent long-term safety performance at costs competitive with mined repositories. Significant fluid flow through basement rock is prevented, in part, by low permeabilities, poorly connected transport pathways, and overburden self-sealing. Deep fluids also resist vertical movement because they are density stratified. Thermal hydrologic calculations estimate the thermal pulse from emplaced waste to be small (less than 20 C at 10 meters from the borehole, for less than a few hundred years), and to result in maximum total vertical fluid movement of {approx}100 m. Reducing conditions will sharply limit solubilities of most dose-critical radionuclides at depth, and high ionic strengths of deep fluids will prevent colloidal transport. For the bounding analysis of this report, waste is envisioned to be emplaced as fuel assemblies stacked inside drill casing that are lowered, and emplaced using off-the-shelf oilfield and geothermal drilling techniques, into the lower 1-2 km portion of a vertical borehole {approx}45 cm in diameter and 3-5 km deep, followed by borehole sealing. Deep borehole disposal of radioactive waste in the United States would require modifications to the Nuclear Waste Policy Act and to applicable regulatory standards for long-term performance set by the US Environmental Protection Agency (40 CFR part 191) and US Nuclear Regulatory Commission (10 CFR part 60). The performance analysis described here is based on the assumption that long-term standards for deep borehole disposal would be identical in the key regards to those prescribed for existing repositories (40 CFR part 197 and 10 CFR part 63).

  9. Identifying High Interest/Low Reading Level Books

    ERIC Educational Resources Information Center

    Bates, Barbara S.

    1977-01-01

    Discussed are criteria for identifying and selecting books written three or more reading levels below interest level. Characteristics of these books are discussed with respect to their appearance, content (fiction and non-fiction), and style. (JPF)

  10. Why consider subseabed disposal of high-level nuclear waste

    SciTech Connect

    Heath, G. R.; Hollister, C. D.; Anderson, D. R.; Leinen, M.

    1980-01-01

    Large areas of the deep seabed warrant assessment as potential disposal sites for high-level radioactive waste because: (1) they are far from seismically and tectonically active lithospheric plate boundaries; (2) they are far from active or young volcanos; (3) they contain thick layers of very uniform fine-grained clays; (4) they are devoid of natural resources likely to be exploited in the forseeable future; (5) the geologic and oceanographic processes governing the deposition of sediments in such areas are well understood, and are remarkably insensitive to past oceanographic and climatic changes; and (6) sedmentary records of tens of millions of years of slow, uninterrupted deposition of fine grained clay support predictions of the future stability of such sites. Data accumulated to date on the permeability, ion-retardation properties, and mechanical strength of pelagic clay sediments indicate that they can act as a primary barrier to the escape of buried nuclides. Work in progress should determine within the current decade whether subseabed disposal is environmentally acceptable and technically feasible, as well as address the legal, political and social issues raised by this new concept.

  11. The ALICE High Level Trigger: status and plans

    NASA Astrophysics Data System (ADS)

    Krzewicki, Mikolaj; Rohr, David; Gorbunov, Sergey; Breitner, Timo; Lehrbach, Johannes; Lindenstruth, Volker; Berzano, Dario

    2015-12-01

    The ALICE High Level Trigger (HLT) is an online reconstruction, triggering and data compression system used in the ALICE experiment at CERN. Unique among the LHC experiments, it extensively uses modern coprocessor technologies like general purpose graphic processing units (GPGPU) and field programmable gate arrays (FPGA) in the data flow. Realtime data compression is performed using a cluster finder algorithm implemented on FPGA boards. These data, instead of raw clusters, are used in the subsequent processing and storage, resulting in a compression factor of around 4. Track finding is performed using a cellular automaton and a Kalman filter algorithm on GPGPU hardware, where both CUDA and OpenCL technologies can be used interchangeably. The ALICE upgrade requires further development of online concepts to include detector calibration and stronger data compression. The current HLT farm will be used as a test bed for online calibration and both synchronous and asynchronous processing frameworks already before the upgrade, during Run 2. For opportunistic use as a Grid computing site during periods of inactivity of the experiment a virtualisation based setup is deployed.

  12. High-level simulation of JWST event-driven operations

    NASA Astrophysics Data System (ADS)

    Henry, R.; Kinzel, W.

    2012-09-01

    The James Webb Space Telescope (JWST) has an event-driven architecture: an onboard Observation Plan Executive (OPE) executes an Observation Plan (OP) consisting of a sequence of observing units (visits). During normal operations, ground action to update the OP is only expected to be necessary about once a week. This architecture is designed to tolerate uncertainty in visit duration, and occasional visit failures due to inability to acquire guide stars, without creating gaps in the observing timeline. The operations concept is complicated by the need for occasional scheduling of timecritical science and engineering visits that cannot tolerate much slippage without inducing gaps, and also by onboard momentum management. A prototype Python tool called the JWST Observation Plan Execution Simulator (JOPES) has recently been developed to simulate OP execution at a high level and analyze the response of the Observatory and OPE to both nominal and contingency scenarios. Incorporating both deterministic and stochastic behavior, JOPES has potential to be a powerful tool for several purposes: requirements analysis, system verification, systems engineering studies, and test data generation. It has already been successfully applied to a study of overhead estimation bias: whether to use conservative or average-case estimates for timing components that are inherently uncertain, such as those involving guide-star acquisition. JOPES is being enhanced to support interfaces to the operational Proposal Planning Subsystem (PPS) now being developed, with the objective of "closing the loop" between testing and simulation by feeding simulated event logs back into the PPS.

  13. Metrics associated with NIH funding: a high-level view

    PubMed Central

    Jordan, Paul

    2011-01-01

    Objective To introduce the availability of grant-to-article linkage data associated with National Institutes of Health (NIH) grants and to perform a high-level analysis of the publication outputs and impacts associated with those grants. Design Articles were linked to the grants they acknowledge using the grant acknowledgment strings in PubMed using a parsing and matching process as embodied in the NIH Scientific Publication Information Retrieval & Evaluation System system. Additional data from PubMed and citation counts from Scopus were added to the linkage data. The data comprise 2 572 576 records from 1980 to 2009. Results The data show that synergies between NIH institutes are increasing over time; 29% of current articles acknowledge grants from multiple institutes. The median time lag to publication for a new grant is 3 years. Each grant contributes to approximately 1.7 articles per year, averaged over all grant types. Articles acknowledging US Public Health Service (PHS, which includes NIH) funding are cited twice as much as US-authored articles acknowledging no funding source. Articles acknowledging both PHS funding and a non-US government funding source receive on average 40% more citations that those acknowledging PHS funding sources alone. Conclusion The US PHS is effective at funding research with a higher-than-average impact. The data are amenable to further and much more detailed analysis. PMID:21527408

  14. Commissioning and first experiences of the ALICE High Level Trigger

    NASA Astrophysics Data System (ADS)

    Steinbeck, Timm M.; Alice Hlt Collaboration

    2010-04-01

    For the ALICE heavy-ion experiment a large computing cluster will be used to perform the last triggering stages in the High Level Trigger (HLT). For the first year of operation the cluster consisted of about 100 multi-processing nodes with 4 or 8 CPU cores each, to be increased to more than 1000 nodes for the coming years of operation. During the commissioning phases of the detector, the preparations for first LHC beam, as well as during the periods of first LHC beam, the HLT has been used extensively already to reconstruct, compress, and display data from the different detectors. For example the HLT has been used to compress Silicon Drift Detector (SDD) data by a factor of 15, lossless, on the fly at a rate of more than 800 Hz. For ALICE's Time Projection Chamber (TPC) detector the HLT has been used to reconstruct tracks online and show the reconstructed tracks in an online event display. The event display can also display online reconstructed data from the Dimuon and Photon Spectrometer (PHOS) detectors. For the latter detector a first selection mechanism has also been put into place to select only events for forwarding to the online display in which data has passed through the PHOS detector. In this contribution we will present experiences and results from these commissioning phases.

  15. High Level Trigger Applications for the ALICE Experiment

    NASA Astrophysics Data System (ADS)

    Richter, M.; Aamodt, K.; Alt, T.; Bablok, S.; Cheshkov, C.; Hille, P. T.; Lindenstruth, V.; Ovrebekk, G.; Ploskon, M.; Popescu, S.; Rohrich, D.; Steinbeck, T. M.; Thader, J.

    2008-02-01

    For the ALICE experiment at the Large Hadron Collider (LHC) at CERN/Geneva, a high level trigger system (HLT) for online event selection and data compression has been developed and a computing cluster of several hundred dual-processor nodes is being installed. A major system integration test was carried out during the commissioning of the time projection chamber (TPC), where the HLT also provides a monitoring system. All major parts like a small computing cluster, hardware input devices, the online data transportation framework, and the HLT analysis could be tested successfully. A common interface for HLT processing components has been designed to run the components from either the online or offline analysis framework without changes. The interface adapts the component to the needs of the online processing and allows the developer to use the offline framework for easy development, debugging, and benchmarking. Following this approach, results can be compared directly. For the upcoming commissioning of the whole detector, the HLT is currently prepared to run online data analysis for the main detectors, e.g., the inner tracking system (ITS), the TPC, and the transition radiation detector (TRD). The HLT processing capability is indispensable for the photon spectrometer (PHOS), where the online pulse shape analysis reduces the data volume by a factor 20. A common monitoring framework is in place and detector calibration algorithms have been ported to the HLT. The paper describes briefly the architecture of the HLT system. It focuses on typical applications and component development.

  16. High Level Waste Feed Certification in Hanford Double Shell Tanks

    SciTech Connect

    Thien, Micheal G.; Wells, Beric E.; Adamson, Duane J.

    2010-03-01

    The ability to effectively mix, sample, certify, and deliver consistent batches of High Level Waste (HLW) feed from the Hanford Double Shell Tanks (DST) to the Waste Treatment and Immobilization Plant (WTP) presents a significant mission risk with potential to impact mission length and the quantity of HLW glass produced. DOE’s River Protection Project (RPP) mission modeling and WTP facility modeling assume that individual 3785 cubic meter (1 million gallon) HLW feed tanks are homogenously mixed, representatively sampled, and consistently delivered to the WTP. It has been demonstrated that homogenous mixing of HLW sludge in Hanford DSTs is not likely achievable with the baseline design thereby causing representative sampling and consistent feed delivery to be more difficult. Inconsistent feed to the WTP could cause additional batch to batch operational adjustments that reduces operating efficiency and has the potential to increase the overall mission length. The Hanford mixing and sampling demonstration program will identify DST mixing performance capability, will evaluate representative sampling techniques, and will estimate feed batch consistency. An evaluation of demonstration program results will identify potential mission improvement considerations that will help ensure successful mission completion. This paper will discuss the history, progress, and future activities that will define and mitigate the mission risk.

  17. A high rate clarifier for load levelling in sewerage systems.

    PubMed

    Jago, R A; Davey, A; Li, H

    2003-01-01

    The combining of chemically assisted clarification with a proprietary physical separation technology has led to a high rate process for clarifying flocculated sewage and other waste streams. This hybrid physico-chemical system, known as the CDS Fine Solids Separation (FSS) System, was developed over a two year period within a sewage treatment plant environment. This paper summarises the results of a recent field trial of the system with a Victorian water authority which experiences heavy loading of sewers in a coastal town during holiday periods. The trial sought to evaluate the FSS as a tool for smoothing the load on the 11 km long sewer to the sewage treatment plant (STP). The FSS system could possibly enable the costly augmentation of the sewer to be deferred, particularly as the capacity of the existing sewer pipe is satisfactory for most of the year. Water quality parameters were determined for a range of flowrates and operational conditions over a two month period. Large reductions were achieved in TSS, TP, FC, turbidity and BOD5, with only minimal reductions in NH3 and TON. These results showed that the FSS could meet the authority's objectives for load levelling and would provide a 20-25% increase in effective sewer capacity. The data are also discussed in terms of possible use of the effluent from the FSS for water reuse applications.

  18. Potential for erosion corrosion of SRS high level waste tanks

    SciTech Connect

    Zapp, P.E.

    1994-01-01

    SRS high-level radioactive waste tanks will not experience erosion corrosion to any significant degree during slurry pump operations. Erosion corrosion in carbon steel structures at reported pump discharge velocities is dominated by electrochemical (corrosion) processes. Interruption of those processes, as by the addition of corrosion inhibitors, sharply reduces the rate of metal loss from erosion corrosion. The well-inhibited SRS waste tanks have a near-zero general corrosion rate, and therefore will be essentially immune to erosion corrosion. The experimental data on carbon steel erosion corrosion most relevant to SRS operations was obtained at the Hanford Site on simulated Purex waste. A metal loss rate of 2.4 mils per year was measured at a temperature of 102 C and a slurry velocity comparable to calculated SRS slurry velocities on ground specimens of the same carbon steel used in SRS waste tanks. Based on these data and the much lower expected temperatures, the metal loss rate of SRS tanks under waste removal and processing conditions should be insignificant, i.e. less than 1 mil per year.

  19. Exercise responses in patients with chronically high creatine kinase levels.

    PubMed

    Cooper, Christopher B; Dolezal, Brett A; Neufeld, Eric V; Shieh, Perry; Jenner, John R; Riley, Marshall

    2017-08-01

    Elevated serum creatine kinase (CK) is often taken to reflect muscle disease, but many individuals have elevated CK without a specific diagnosis. How elevated CK reflects muscle metabolism during exercise is not known. Participants (46 men, 48 women) underwent incremental exercise testing to assess aerobic performance, cardiovascular response, and ventilatory response. Serum lactate, ammonia, and CK were measured at rest, 4 minutes into exercise, and 2 minutes into recovery. High-CK and control subjects demonstrated similar aerobic capacities and cardiovascular responses to incremental exercise. Those with CK ≥ 300 U/L exhibited significantly higher lactate and ammonia levels after maximal exercise, together with increased ventilatory responses, whereas those with CK ≥200 U/L but ≤ 300 U/L did not. We recommend measurement of lactate and ammonia profiles during a maximal incremental exercise protocol to help identify patients who warrant muscle biopsy to rule out myopathy. Muscle Nerve 56: 264-270, 2017. © 2016 Wiley Periodicals, Inc.

  20. Identification of areas with high levels of untreated dental caries.

    PubMed

    Ellwood, R P; O'Mullane, D M

    1996-02-01

    In order to examine the geographical variation of dental health within 10 county districts in North Wales, 3538 children were examined. The associations between three demographic indicators, based on the 1981 OPCS census, and dental health outcomes were assessed for electoral wards within the county districts. The Townsend and Jarman indices were the first two indicators employed and the third was based on a mathematical model representing the variation in the mean number of untreated decayed surfaces per person for the wards. This model was developed using the children examined in the five most westerly county districts. Using the data derived from the five most easterly county districts, the three indicators were assessed. All three showed strong correlations (r > or = 0.88) with dental health. These results indicate that measures of dental health based on large administrative units may obscure variation within them. It is concluded that geographical methods of this type may be useful for targeting dental resources at small areas with high levels of need.

  1. High-Level Performance Modeling of SAR Systems

    NASA Technical Reports Server (NTRS)

    Chen, Curtis

    2006-01-01

    SAUSAGE (Still Another Utility for SAR Analysis that s General and Extensible) is a computer program for modeling (see figure) the performance of synthetic- aperture radar (SAR) or interferometric synthetic-aperture radar (InSAR or IFSAR) systems. The user is assumed to be familiar with the basic principles of SAR imaging and interferometry. Given design parameters (e.g., altitude, power, and bandwidth) that characterize a radar system, the software predicts various performance metrics (e.g., signal-to-noise ratio and resolution). SAUSAGE is intended to be a general software tool for quick, high-level evaluation of radar designs; it is not meant to capture all the subtleties, nuances, and particulars of specific systems. SAUSAGE was written to facilitate the exploration of engineering tradeoffs within the multidimensional space of design parameters. Typically, this space is examined through an iterative process of adjusting the values of the design parameters and examining the effects of the adjustments on the overall performance of the system at each iteration. The software is designed to be modular and extensible to enable consideration of a variety of operating modes and antenna beam patterns, including, for example, strip-map and spotlight SAR acquisitions, polarimetry, burst modes, and squinted geometries.

  2. High resolution LES study of the nocturnal low level jet

    NASA Astrophysics Data System (ADS)

    Giometto, Marco; Calaf, Marc; Oldroyd, Holly; Fang, Jiannong; Parlange, Marc B.

    2013-04-01

    Katabatic winds are buoyantly driven flows arising along cooled sloping surfaces which play a crucial role in driving the local weather, redistributing scalars such as temperature and moisture in the atmosphere. These winds are established following sunset under strong radiational cooling and rapidly stop after dawn with the formation of the convective boundary layer. They are characterized by a peak in the along slope velocity known as nocturnal low level jet (LLJ) whose effects, on the dynamics of such systems, have been recently investigated but are still not fully understood. The current contribution proposes a Large Eddy Simulation (LES) study at high resolution of idealized katabatic flows along cooled sloping surfaces and aims at gaining a deeper understanding on those that are the dynamics of such thermodynamical systems at the LLJ height. The stably stratified atmosphere is approximated in the Boussinesq sense, rotational effects are not taken into account and the subgrid terms for momentum and buoyancy are independently parametrized adopting Lagrangian scale dependent dynamic models (Bou Zeid et al., 2005). The structure of the mean and turbulent fields obtained from our numerical setup is analysed and results are compared with recent literature and meteorological observations from a narrow alpine valley with steep slopes (Val Ferret, Switzerland). The importance of the subgrid parametrization is tested via run at various resolution.

  3. Defense High-Level Waste Leaching Mechanisms Program. Final report

    SciTech Connect

    Mendel, J.E.

    1984-08-01

    The Defense High-Level Waste Leaching Mechanisms Program brought six major US laboratories together for three years of cooperative research. The participants reached a consensus that solubility of the leached glass species, particularly solubility in the altered surface layer, is the dominant factor controlling the leaching behavior of defense waste glass in a system in which the flow of leachant is constrained, as it will be in a deep geologic repository. Also, once the surface of waste glass is contacted by ground water, the kinetics of establishing solubility control are relatively rapid. The concentrations of leached species reach saturation, or steady-state concentrations, within a few months to a year at 70 to 90/sup 0/C. Thus, reaction kinetics, which were the main subject of earlier leaching mechanisms studies, are now shown to assume much less importance. The dominance of solubility means that the leach rate is, in fact, directly proportional to ground water flow rate. Doubling the flow rate doubles the effective leach rate. This relationship is expected to obtain in most, if not all, repository situations.

  4. EUVE GO Survey: High Levels of User Satisfaction

    NASA Astrophysics Data System (ADS)

    Stroozas, B. A.

    2000-12-01

    This paper describes the results of a detailed customer survey of Guest Observers (GOs) for NASA's Extreme Ultraviolet Explorer (EUVE) astronomy satellite observatory. The purpose of the research survey was to (1) measure the levels of GO customer satisfaction with respect to EUVE observing services, and (2) compare the observing experiences of EUVE GOs with their experiences using other satellite observatories. This survey was conducted as a business research project -- part of the author's graduate work as an MBA candidate. A total sample of 38 respondents, from a working population of 101 "active" EUVE GOs, participated in this survey. The results, which provided a profile of the "typical" EUVE GO, showed in a statistically significant fashion that these GOs were more than satisfied with the available EUVE observing services. In fact, the sample GOs generally rated their EUVE observing experiences to be better than average as compared to their experiences as GOs on other missions. These relatively high satisfaction results are particularly pleasing to the EUVE Project which, given its significantly reduced staffing environment at U.C. Berkeley, has continued to do more with less. This paper outlines the overall survey process: the relevant background and previous research, the survey design and methodology, and the final results and their interpretation. The paper also points out some general limitations and weaknesses of the study, along with some recommended actions for the EUVE Project and for NASA in general. This work was funded by NASA/UCB Cooperative Agreement NCC5-138.

  5. The GRAVITY instrument software/high-level software

    NASA Astrophysics Data System (ADS)

    Burtscher, Leonard; Wieprecht, Ekkehard; Ott, Thomas; Kok, Yitping; Yazici, Senol; Anugu, Narsireddy; Dembet, Roderick; Fedou, Pierre; Lacour, Sylvestre; Ott, Jürgen; Paumard, Thibaut; Lapeyrere, Vincent; Kervella, Pierre; Abuter, Roberto; Pozna, Eszter; Eisenhauer, Frank; Blind, Nicolas; Genzel, Reinhard; Gillessen, Stefan; Hans, Oliver; Haug, Marcus; Haussmann, Frank; Kellner, Stefan; Lippa, Magdalena; Pfuhl, Oliver; Sturm, Eckhard; Weber, Johannes; Amorim, Antonio; Brandner, Wolfgang; Rousselet-Perraut, Karine; Perrin, Guy S.; Straubmeier, Christian; Schöller, Markus

    2014-07-01

    GRAVITY is the four-beam, near-infrared, AO-assisted, fringe tracking, astrometric and imaging instrument for the Very Large Telescope Interferometer (VLTI). It is requiring the development of one of the most complex instrument software systems ever built for an ESO instrument. Apart from its many interfaces and interdependencies, one of the most challenging aspects is the overall performance and stability of this complex system. The three infrared detectors and the fast reflective memory network (RMN) recorder contribute a total data rate of up to 20 MiB/s accumulating to a maximum of 250 GiB of data per night. The detectors, the two instrument Local Control Units (LCUs) as well as the five LCUs running applications under TAC (Tools for Advanced Control) architecture, are interconnected with fast Ethernet, RMN fibers and dedicated fiber connections as well as signals for the time synchronization. Here we give a simplified overview of all subsystems of GRAVITY and their interfaces and discuss two examples of high-level applications during observations: the acquisition procedure and the gathering and merging of data to the final FITS file.

  6. Cytotoxicity assessment of residual high-level disinfectants.

    PubMed

    Ryu, Mizuyuki; Kobayashi, Toshihiro; Kawamukai, Emiko; Quan, Glenlelyn; Furuta, Taro

    2013-01-01

    Some studies show the uptake of disinfectants on medical devices but no studies on their cytotoxicity have been reported. This study aimed to assess that cytotoxicity in a 3-dimensional culture system using HeLa cells grown in matrices composed of collagen. Plastic materials were soaked in the use solutions of the widely used high-level disinfectants, glutaraldehyde (GA), ortho-phthalaldehyde (OPA) and peracetic acid (PAA). After being rinsed, they were allowed to dry and were embedded into the cell medium to investigate the cytotoxicity of the residual disinfectants. Cytotoxicity was observed with the polyvinyl chloride, polyurethane and silicon tubes soaked in GA and OPA, indicating that both disinfectants were absorbed in the test pieces, whereas for PAA, none was observed. As for the polytetrafluoroethylene (PTFE) tubes, no disinfectant displayed cytotoxicity. GA and OPA are primary irritants, having a potential to cause anaphylaxis and other forms of allergic reactions. There should be consideration not only about the toxicity of the residual disinfectant from poor rinsing, but also about the toxicity that would result from the disinfectants that were absorbed and consequently released from the medical devices or materials.

  7. The IFR pyroprocessing for high-level waste minimization

    SciTech Connect

    Laidler, J.J. )

    1993-01-01

    The process developed for the recycle of integral fast reactor (IFR) spent fuel utilizes a combination of pyrometallurgical and electrochemical methods and has been termed pyroprocessing. The process has been operated at full scale with simulated spent fuel using nonradioactive fission product elements. A comprehensive demonstration of the pyroprocessing of irradiated IFR fuel will begin later this year. Pyroprocessing involves the anodic dissolution of all the constituent elements of the IFR spent fuel and controlled electrotransport (electrorefining) to separate the actinide elements from the fission products present in the spent fuel. The process be applied to the processing of spent light water reactor (LWR) fuel as well, requiring only the addition of a reduction step to convert the LWR fuel as well, requiring only the addition of a reduction step to convert the LWR oxide fuel to metallic form and a separation step to separate uranium from the transuranic (TRU) elements. The TRU elements are then recovered by electroefining in the same manner as the actinides from the IFR high-level wastes arising from pyroprocessing are virtually free of actinides, and the volume of the wastes is minimized by the intrinsic characteristics of the processing of the processing method.

  8. Burning experiments and late Paleozoic high O2 levels

    NASA Astrophysics Data System (ADS)

    Wildman, R.; Essenhigh, R.; Berner, R.; Hickey, L.; Wildman, C.

    2003-04-01

    The Paleozoic rise of land plants brought about increased burial of organic matter and a resulting increase in atmospheric oxygen concentrations. Levels as high as 30-35% O2 may have been reached during the Permo-Carboniferous (Berner and Canfield, 1989; Berner, 2001). However, burning experiments based solely on paper (Watson, 1978) have challenged these results, the claim being that if the oxygen made up more than 25% of the atmosphere, the frequency and intensity of forest fires would increase sufficiently to prevent the continued existence of plant life. Thus, since plants have persisted, it is possible that fires served as a negative feedback against excessive oxygen levels. An initial study of Paleozoic wildfire behavior via thermogravimetric analysis (TGA) was conducted under ambient and enriched oxygen conditions to simulate present and ancient atmospheres. The tests focused on natural fuels, specifically tree leaves and wood, tree fern fibers, and sphagnum peat-moss, simulating Permo-Carboniferous upland and swampland ecosystems, respectively. Three conclusions are: (1) enriched oxygen increases the rate of mass loss during burning; (2) fuel chemistry (cellulose vs. lignin) influences burning patterns; and (3) in geometrically heterogeneous fuels, geometry affects burning rate significantly. Both geometrically and chemically, paper resists fire poorly; thus, we found that it loses its mass at lower temperatures than forest materials and is therefore a poor proxy for Paleozoic ecosystems. Further study of Paleozoic wildfire spread behavior is currently being conducted. Fires are lit using pine dowels, which allow for reproducible fuel density. Steady-state, one-dimensional flame-spread is measured with thermocouples anchored two inches above the fuel bed. Both oxygen concentration of the air supply to the fire and moisture content of the fuels are varied, as we suspect that these are two main controls of wildfire spread. Burning fuels of varying moisture

  9. Compensatory Changes in the Cytoplasmic Tail of gp41 Confer Resistance to Tetherin/BST-2 in a Pathogenic Nef-deleted SIV

    PubMed Central

    Serra-Moreno, Ruth; Jia, Bin; Breed, Matthew; Alvarez, Xavier; Evans, David T.

    2011-01-01

    SUMMARY Tetherin (BST-2 or CD317) is an interferon-inducible transmembrane protein that inhibits virus release from infected cells. Whereas HIV-1 Vpu and HIV-2 Env counteract human tetherin, most SIVs use Nef to antagonize the tetherin proteins of their non-human primate hosts. Here we show that compensatory changes in the cytoplasmic domain of SIV gp41, acquired by a nef-deleted virus that regained a pathogenic phenotype in infected rhesus macaques, restore resistance to tetherin. These changes facilitate virus release in the presence of rhesus tetherin, but not human tetherin, and enhance virus replication in interferon-treated primary lymphocytes. The substitutions in gp41 result in a selective physical association with rhesus tetherin, and the internalization and sequestration of rhesus tetherin by a mechanism that depends on a conserved endocytosis motif in gp41. These results are consistent with HIV-2 Env antagonism of human tetherin, and suggest that the ability to oppose tetherin is important for lentiviral pathogenesis. PMID:21238946

  10. Oasis: A high-level/high-performance open source Navier-Stokes solver

    NASA Astrophysics Data System (ADS)

    Mortensen, Mikael; Valen-Sendstad, Kristian

    2015-03-01

    Oasis is a high-level/high-performance finite element Navier-Stokes solver written from scratch in Python using building blocks from the FEniCS project (fenicsproject.org). The solver is unstructured and targets large-scale applications in complex geometries on massively parallel clusters. Oasis utilizes MPI and interfaces, through FEniCS, to the linear algebra backend PETSc. Oasis advocates a high-level, programmable user interface through the creation of highly flexible Python modules for new problems. Through the high-level Python interface the user is placed in complete control of every aspect of the solver. A version of the solver, that is using piecewise linear elements for both velocity and pressure, is shown to reproduce very well the classical, spectral, turbulent channel simulations of Moser et al. (1999). The computational speed is strongly dominated by the iterative solvers provided by the linear algebra backend, which is arguably the best performance any similar implicit solver using PETSc may hope for. Higher order accuracy is also demonstrated and new solvers may be easily added within the same framework.

  11. JET MIXING ANALYSIS FOR SRS HIGH-LEVEL WASTE RECOVERY

    SciTech Connect

    Lee, S.

    2011-07-05

    The process of recovering the waste in storage tanks at the Savannah River Site (SRS) typically requires mixing the contents of the tank to ensure uniformity of the discharge stream. Mixing is accomplished with one to four slurry pumps located within the tank liquid. The slurry pump may be fixed in position or they may rotate depending on the specific mixing requirements. The high-level waste in Tank 48 contains insoluble solids in the form of potassium tetraphenyl borate compounds (KTPB), monosodium titanate (MST), and sludge. Tank 48 is equipped with 4 slurry pumps, which are intended to suspend the insoluble solids prior to transfer of the waste to the Fluidized Bed Steam Reformer (FBSR) process. The FBSR process is being designed for a normal feed of 3.05 wt% insoluble solids. A chemical characterization study has shown the insoluble solids concentration is approximately 3.05 wt% when well-mixed. The project is requesting a Computational Fluid Dynamics (CFD) mixing study from SRNL to determine the solids behavior with 2, 3, and 4 slurry pumps in operation and an estimate of the insoluble solids concentration at the suction of the transfer pump to the FBSR process. The impact of cooling coils is not considered in the current work. The work consists of two principal objectives by taking a CFD approach: (1) To estimate insoluble solids concentration transferred from Tank 48 to the Waste Feed Tank in the FBSR process and (2) To assess the impact of different combinations of four slurry pumps on insoluble solids suspension and mixing in Tank 48. For this work, several different combinations of a maximum of four pumps are considered to determine the resulting flow patterns and local flow velocities which are thought to be associated with sludge particle mixing. Two different elevations of pump nozzles are used for an assessment of the flow patterns on the tank mixing. Pump design and operating parameters used for the analysis are summarized in Table 1. The baseline

  12. Millimeter-Wave Measurements of High Level and Low Level Activity Glass Melts

    SciTech Connect

    Woskov, Paul P.; Sundaram, S.K.; Daniel, William E., Jr.

    2006-06-01

    The primary objectives of the current research is to develop on-line sensors for characterizing molten glass in high-level and low-activity waste glass melters using millimeter-wave (MMW) technology and to use this technology to do novel research of melt dynamics. Existing and planned waste glass melters lack sophisticated diagnostics due to the hot, corrosive, and radioactive melter environments. Without process control diagnostics, the Defense Waste Processing Facility (DWPF) and the Waste Treatment Plant (WTP) under construction at Hanford operate by a feed forward process control scheme that relies on predictive models with large uncertainties. This scheme severely limits production throughput and waste loading. Also operations at DWPF have shown susceptibility to anomalies such as pouring, foaming, and combustion gas build up, which can seriously disrupt operations. Future waste chemistries will be even more challenging. The scientific goals of this project are to develop new reliable on-line monitoring capability for important glass process parameters such as temperature profiles, emissivity, density, viscosity, and other characteristics using the unique advantages of millimeter wave electromagnetic radiation that can be eventually implemented in the operating melters. Once successfully developed and implemented, significant cost savings would be realized in melter operations by increasing production through put, reduced storage volumes (through higher waste loading), and reduced risks (prevention or mitigation of anomalies).

  13. Isolation of high quality RNA from cereal seeds containing high levels of starch.

    PubMed

    Wang, Guifeng; Wang, Gang; Zhang, Xiaowei; Wang, Fang; Song, Rentao

    2012-01-01

    Cereals are an important source of food, feed and fuel with a rapidly increasing global demand. However, cereal seeds contain high levels of starch and polysaccharides, making the isolation of high quality RNA extremely difficult. To develop a novel method for extracting high quality total RNA from various starch- and polysaccharides-rich cereal seeds, such as maize, rice, sorghum and wheat. We developed a modified sodium dodecyl sulphate (SDS)/TRIzol method. The combined use of a Tris buffer (pH 9.0) and SDS before TRIzol extraction effectively resolved the problem of seed homogenate solidification in such a buffer. A high concentration of SDS was used separately, not only to promote cell lysis but also to effectively dissolve seed sample containing high levels of starch. Moreover, acid phenol saturated with 0.1  M citrate buffer (pH 4.3) was used to separate RNA from DNAs, proteins and high levels of starch. This rapid protocol was compared with other RNA isolation methods preferentially used for plants rich in polysaccharides and secondary metabolites. Gel electrophoresis analysis indicated that the extracted total RNA had good integrity without apparent DNA contamination. Furthermore, an A₂₆₀/₂₈₀ ratio of approximately 2.0, an A₂₆₀/₂₃₀ ratio of more than 2.0 and RIN values of more than 8.6 indicated that the isolated RNA was of high purity. The isolated RNA was suitable for subsequent molecular manipulations, such as reverse-transcription polymerase chain reaction (PCR), rapid amplification of cDNA ends (RACE) and real-time PCR. The study has described an easy, efficient and highly reproducible method for RNA isolation from various cereal seeds. Copyright © 2011 John Wiley & Sons, Ltd.

  14. Electric Grid Expansion Planning with High Levels of Variable Generation

    SciTech Connect

    Hadley, Stanton W.; You, Shutang; Shankar, Mallikarjun; Liu, Yilu

    2016-02-01

    Renewables are taking a large proportion of generation capacity in U.S. power grids. As their randomness has increasing influence on power system operation, it is necessary to consider their impact on system expansion planning. To this end, this project studies the generation and transmission expansion co-optimization problem of the US Eastern Interconnection (EI) power grid with a high wind power penetration rate. In this project, the generation and transmission expansion problem for the EI system is modeled as a mixed-integer programming (MIP) problem. This study analyzed a time series creation method to capture the diversity of load and wind power across balancing regions in the EI system. The obtained time series can be easily introduced into the MIP co-optimization problem and then solved robustly through available MIP solvers. Simulation results show that the proposed time series generation method and the expansion co-optimization model and can improve the expansion result significantly after considering the diversity of wind and load across EI regions. The improved expansion plan that combines generation and transmission will aid system planners and policy makers to maximize the social welfare. This study shows that modelling load and wind variations and diversities across balancing regions will produce significantly different expansion result compared with former studies. For example, if wind is modeled in more details (by increasing the number of wind output levels) so that more wind blocks are considered in expansion planning, transmission expansion will be larger and the expansion timing will be earlier. Regarding generation expansion, more wind scenarios will slightly reduce wind generation expansion in the EI system and increase the expansion of other generation such as gas. Also, adopting detailed wind scenarios will reveal that it may be uneconomic to expand transmission networks for transmitting a large amount of wind power through a long distance

  15. Qualification of Innovative High Level Waste Pipeline Unplugging Technologies

    SciTech Connect

    McDaniel, D.; Gokaltun, S.; Varona, J.; Awwad, A.; Roelant, D.; Srivastava, R.

    2008-07-01

    In the past, some of the pipelines have plugged during high level waste (HLW) transfers resulting in schedule delays and increased costs. Furthermore, pipeline plugging has been cited by the 'best and brightest' technical review as one of the major issues that can result in unplanned outages at the Waste Treatment Plant causing inconsistent operation. As the DOE moves toward a more active high level waste retrieval, the site engineers will be faced with increasing cross-site pipeline waste slurry transfers that will result in increased probability of a pipeline getting plugged. Hence, availability of a pipeline unplugging tool/technology is crucial to ensure smooth operation of the waste transfers and in ensuring tank farm cleanup milestones are met. FIU had earlier tested and evaluated various unplugging technologies through an industry call. Based on mockup testing, two technologies were identified that could withstand the rigors of operation in a radioactive environment and with the ability to handle sharp 90 elbows. We present results of the second phase of detailed testing and evaluation of pipeline unplugging technologies and the objective is to qualify these pipeline unplugging technologies for subsequent deployment at a DOE facility. The current phase of testing and qualification comprises of a heavily instrumented 3-inch diameter (full-scale) pipeline facilitating extensive data acquisition for design optimization and performance evaluation, as it applies to three types of plugs atypical of the DOE HLW waste. Furthermore, the data from testing at three different lengths of pipe in conjunction with the physics of the process will assist in modeling the unplugging phenomenon that will then be used to scale-up process parameters and system variables for longer and site typical pipe lengths, which can extend as much as up to 19,000 ft. Detailed information resulting from the testing will provide the DOE end-user with sufficient data and understanding of the

  16. Stability of High-Level Radioactive Waste Forms

    SciTech Connect

    Besmann, T.M.

    2001-06-22

    High-level waste (HLW) glass compositions, processing schemes, limits on waste content, and corrosion/dissolution release models are dependent on an accurate knowledge of melting temperatures and thermochemical values. Unfortunately, existing models for predicting these temperatures are empirically-based, depending on extrapolations of experimental information. In addition, present models of leaching behavior of glass waste forms use simplistic assumptions or experimentally measured values obtained under non-realistic conditions. There is thus a critical need for both more accurate and more widely applicable models for HLW glass behavior, which this project addressed. Significant progress was made in this project on modeling HLW glass. Borosilicate glass was accurately represented along with the additional important components that contain iron, lithium, potassium, magnesium, and calcium. The formation of crystalline inclusions in the glass, an issue in Hanford HLW formulations, was modeled and shown to be predictive. Thus the results of this work have already demonstrated practical benefits with the ability to map compositional regions where crystalline material forms, and therefore avoid that detrimental effect. With regard to a fundamental understanding, added insights on the behavior of the components of glass have been obtained, including the potential formation of molecular clusters. The EMSP project had very significant effects beyond the confines of Environmental Management. The models developed for glass have been used to solve a very costly problem in the corrosion of refractories for glass production. The effort resulted in another laboratory, Sandia National Laboratories-Livermore, to become conversant in the techniques and to apply those through a DOE Office of Industrial Technologies project joint with PPG Industries. The glass industry as a whole is now cognizant of these capabilities, and there is a Glass Manufacturer's Research Institute proposal

  17. Seasonal changes in stress indicators in high level football.

    PubMed

    Faude, O; Kellmann, M; Ammann, T; Schnittker, R; Meyer, T

    2011-04-01

    This study aimed at describing changes in stress and performance indicators throughout a competitive season in high level football. 15 players (19.5±3.0 years, 181±5 cm, 75.7±9.0 kg) competing under professional circumstances were tested at baseline and 3 times during the season 2008/09 (in-season 1, 2, 3). Testing consisted of the Recovery-Stress Questionnaire for Athletes (Total Stress and Recovery score), vertical jump tests (counter movement and drop jump (DJ)), and a maximal ramp-like running test. Average match exposure was higher during a 3-weeks period prior to in-season 3 compared to in-season 1 and 2 (1.5 vs. 1 h/week, p=0.05). Total Stress score was elevated at in-season 1 and 2 compared to baseline (p<0.01) with a further increase at in-season 3 (p<0.03; generalized eta squared (η(2)(g))=0.37). Total Recovery score was decreased at in-season 1 and 3 compared to baseline (p<0.05; η(2)(g)=0.21). Maximal running velocity (V(max)) and jumping heights were not significantly affected (η(2)(g)≤0.04). Changes in DJ height and V (max) between baseline and in-season 3 were correlated with the corresponding changes in Total Stress score (r=-0.55 and r=-0.61, p<0.03). Usual match exposure during a professional football season does not induce relevant changes in performance indicators. Accumulated stress and a lack of recovery towards the end of a season might be indicated by psychometric deteriorations. © Georg Thieme Verlag KG Stuttgart · New York.

  18. High-Level Waste Systems Plan. Revision 7

    SciTech Connect

    Brooke, J.N.; Gregory, M.V.; Paul, P.; Taylor, G.; Wise, F.E.; Davis, N.R.; Wells, M.N.

    1996-10-01

    This revision of the High-Level Waste (HLW) System Plan aligns SRS HLW program planning with the DOE Savannah River (DOE-SR) Ten Year Plan (QC-96-0005, Draft 8/6), which was issued in July 1996. The objective of the Ten Year Plan is to complete cleanup at most nuclear sites within the next ten years. The two key principles of the Ten Year Plan are to accelerate the reduction of the most urgent risks to human health and the environment and to reduce mortgage costs. Accordingly, this System Plan describes the HLW program that will remove HLW from all 24 old-style tanks, and close 20 of those tanks, by 2006 with vitrification of all HLW by 2018. To achieve these goals, the DWPF canister production rate is projected to climb to 300 canisters per year starting in FY06, and remain at that rate through the end of the program in FY18, (Compare that to past System Plans, in which DWPF production peaked at 200 canisters per year, and the program did not complete until 2026.) An additional $247M (FY98 dollars) must be made available as requested over the ten year planning period, including a one-time $10M to enhance Late Wash attainment. If appropriate resources are made available, facility attainment issues are resolved and regulatory support is sufficient, then completion of the HLW program in 2018 would achieve a $3.3 billion cost savings to DOE, versus the cost of completing the program in 2026. Facility status information is current as of October 31, 1996.

  19. High-level waste issues and resolutions document

    SciTech Connect

    Not Available

    1994-05-01

    The High-Level Waste (HLW) Issues and Resolutions Document recognizes US Department of Energy (DOE) complex-wide HLW issues and offers potential corrective actions for resolving these issues. Westinghouse Management and Operations (M&O) Contractors are effectively managing HLW for the Department of Energy at four sites: Idaho National Engineering Laboratory (INEL), Savannah River Site (SRS), West Valley Demonstration Project (WVDP), and Hanford Reservation. Each site is at varying stages of processing HLW into a more manageable form. This HLW Issues and Resolutions Document identifies five primary issues that must be resolved in order to reach the long-term objective of HLW repository disposal. As the current M&O contractor at DOE`s most difficult waste problem sites, Westinghouse recognizes that they have the responsibility to help solve some of the complexes` HLW problems in a cost effective manner by encouraging the M&Os to work together by sharing expertise, eliminating duplicate efforts, and sharing best practices. Pending an action plan, Westinghouse M&Os will take the initiative on those corrective actions identified as the responsibility of an M&O. This document captures issues important to the management of HLW. The proposed resolutions contained within this document set the framework for the M&Os and DOE work cooperatively to develop an action plan to solve some of the major complex-wide problems. Dialogue will continue between the M&Os, DOE, and other regulatory agencies to work jointly toward the goal of storing, treating, and immobilizing HLW for disposal in an environmentally sound, safe, and cost effective manner.

  20. PLUTONIUM/HIGH-LEVEL VITRIFIED WASTE BDBE DOSE CALCULATION

    SciTech Connect

    D.C. Richardson

    2003-03-19

    In accordance with the Nuclear Waste Policy Amendments Act of 1987, Yucca Mountain was designated as the site to be investigated as a potential repository for the disposal of high-level radioactive waste. The Yucca Mountain site is an undeveloped area located on the southwestern edge of the Nevada Test Site (NTS), about 100 miles northwest of Las Vegas. The site currently lacks rail service or an existing right-of-way. If the Yucca Mountain site is found suitable for the repository, rail service is desirable to the Office of Civilian Waste Management (OCRWM) Program because of the potential of rail transportation to reduce costs and to reduce the number of shipments relative to highway transportation. A Preliminary Rail Access Study evaluated 13 potential rail spur options. Alternative routes within the major options were also developed. Each of these options was then evaluated for potential land use conflicts and access to regional rail carriers. Three potential routes having few land use conflicts and having access to regional carriers were recommended for further investigation. Figure 1-1 shows these three routes. The Jean route is estimated to be about 120 miles long, the Carlin route to be about 365 miles long, and Caliente route to be about 365 miles long. The remaining ten routes continue to be monitored and should any of the present conflicts change, a re-evaluation of that route will be made. Complete details of the evaluation of the 13 routes can be found in the previous study. The DOE has not identified any preferred route and recognizes that the transportation issues need a full and open treatment under the National Environmental Policy Act. The issue of transportation will be included in public hearings to support development of the Environmental Impact Statement (EIS) proceedings for either the Monitored Retrievable Storage Facility or the Yucca Mountain Project or both.

  1. Level 1 Tornado PRA for the High Flux Beam Reactor

    SciTech Connect

    Bozoki, G.E.; Conrad, C.S.

    1994-05-01

    This report describes a risk analysis primarily directed at providing an estimate for the frequency of tornado induced damage to the core of the High Flux Beam Reactor (HFBR), and thus it constitutes a Level 1 Probabilistic Risk Assessment (PRA) covering tornado induced accident sequences. The basic methodology of the risk analysis was to develop a ``tornado specific`` plant logic model that integrates the internal random hardware failures with failures caused externally by the tornado strike and includes operator errors worsened by the tornado modified environment. The tornado hazard frequency, as well as earlier prepared structural and equipment fragility data, were used as input data to the model. To keep modeling/calculational complexity as simple as reasonable a ``bounding`` type, slightly conservative, approach was applied. By a thorough screening process a single dominant initiating event was selected as a representative initiator, defined as: ``Tornado Induced Loss of Offsite Power.`` The frequency of this initiator was determined to be 6.37E-5/year. The safety response of the HFBR facility resulted in a total Conditional Core Damage Probability of .621. Thus, the point estimate of the HFBR`s Tornado Induced Core Damage Frequency (CDF) was found to be: (CDF){sub Tornado} = 3.96E-5/year. This value represents only 7.8% of the internal CDF and thus is considered to be a small contribution to the overall facility risk expressed in terms of total Core Damage Frequency. In addition to providing the estimate of (CDF){sub Tornado}, the report documents, the relative importance of various tornado induced system, component, and operator failures that contribute most to (CDF){sub Tornado}.

  2. Do mothers with high sodium levels in their breast milk have high depression and anxiety scores?

    PubMed

    Serim Demirgoren, Burcu; Ozbek, Aylin; Ormen, Murat; Kavurma, Canem; Ozer, Esra; Aydın, Adem

    2017-04-01

    Objective This study aimed to assess the possible association of high breast milk sodium levels with postpartum depression and anxiety. Methods A total of 150 mothers and their healthy, exclusively breastfed newborns aged 8 to 15 days were recruited. Mothers were asked to complete scales for evaluation of postnatal depression and anxiety following an interview for consent and sociodemographic data collection. Breast milk samples were obtained to measure sodium and potassium (K) levels. Results Forty-nine mothers had higher than expected breast milk Na concentrations and a high Na/K ratio. These mothers scored significantly higher on the scales of postnatal depression and state anxiety ( P = 0.018 and P = 0.048, respectively). Conclusions This study shows that compared to normal breast milk Na levels and Na/K ratio, high breast milk Na and high Na/K ratio, with possible serious consequences in infants, are associated with maternal depressive and anxious symptoms in the postpartum period.

  3. Concerns--High Sea Levels and Temperatures Seen Next Century.

    ERIC Educational Resources Information Center

    Ryan, Paul R.

    1984-01-01

    A National Research Council committee recently concluded that atmospheric carbon dioxide levels will "most likely" double by late in the next century, causing an increase in the earth's average temperature. Effects of the increase on sea levels, global climate, and other parameters are discussed. (JN)

  4. Concerns--High Sea Levels and Temperatures Seen Next Century.

    ERIC Educational Resources Information Center

    Ryan, Paul R.

    1984-01-01

    A National Research Council committee recently concluded that atmospheric carbon dioxide levels will "most likely" double by late in the next century, causing an increase in the earth's average temperature. Effects of the increase on sea levels, global climate, and other parameters are discussed. (JN)

  5. Confidence Level and Sensitivity Limits in High-Contrast Imaging

    NASA Astrophysics Data System (ADS)

    Marois, Christian; Lafrenière, David; Macintosh, Bruce; Doyon, René

    2008-01-01

    In long adaptive optics corrected exposures, exoplanet detections are currently limited by speckle noise originating from the telescope and instrument optics, and it is expected that such noise will also limit future high-contrast imaging instruments for both ground- and space-based telescopes. Previous theoretical analyses have shown that the time intensity variations of a single speckle follow a modified Rician. It is first demonstrated here that for a circular pupil, this temporal intensity distribution also represents the speckle spatial intensity distribution at a fixed separation from the point-spread function center; this fact is demonstrated using numerical simulations for coronagraphic and noncoronagraphic data. The real statistical distribution of the noise needs to be taken into account explicitly when selecting a detection threshold appropriate for some desired confidence level (CL). In this paper, a technique is described to obtain the pixel intensity distribution of an image and its corresponding CL as a function of the detection threshold. Using numerical simulations, it is shown that in the presence of speckle noise, a detection threshold up to 3 times higher is required to obtain a CL equivalent to that at 5 σ for Gaussian noise. The technique is then tested on data acquired by simultaneous spectral differential imaging with TRIDENT and by angular differential imaging with NIRI. It is found that the angular differential imaging technique produces quasi-Gaussian residuals, a remarkable result compared to classical adaptive optic imaging. Finally, a power law is derived to predict the 1 - 3 × 10-7 CL detection threshold when averaging a partially correlated non-Gaussian noise. Based on observations obtained at the Canada-France-Hawaii Telescope (CFHT), which is operated by the National Research Council of Canada, the Institut National des Sciences de l'Univers of the Centre National de la Recherche Scientifique of France, and the University of

  6. A high-speed DAQ framework for future high-level trigger and event building clusters

    NASA Astrophysics Data System (ADS)

    Caselle, M.; Ardila Perez, L. E.; Balzer, M.; Dritschler, T.; Kopmann, A.; Mohr, H.; Rota, L.; Vogelgesang, M.; Weber, M.

    2017-03-01

    Modern data acquisition and trigger systems require a throughput of several GB/s and latencies of the order of microseconds. To satisfy such requirements, a heterogeneous readout system based on FPGA readout cards and GPU-based computing nodes coupled by InfiniBand has been developed. The incoming data from the back-end electronics is delivered directly into the internal memory of GPUs through a dedicated peer-to-peer PCIe communication. High performance DMA engines have been developed for direct communication between FPGAs and GPUs using "DirectGMA (AMD)" and "GPUDirect (NVIDIA)" technologies. The proposed infrastructure is a candidate for future generations of event building clusters, high-level trigger filter farms and low-level trigger system. In this paper the heterogeneous FPGA-GPU architecture will be presented and its performance be discussed.

  7. High-Dimensional Topological Insulators with Quaternionic Analytic Landau Levels

    NASA Astrophysics Data System (ADS)

    Li, Yi; Wu, Congjun

    2013-05-01

    We study the three-dimensional topological insulators in the continuum by coupling spin-1/2 fermions to the Aharonov-Casher SU(2) gauge field. They exhibit flat Landau levels in which orbital angular momentum and spin are coupled with a fixed helicity. The three-dimensional lowest Landau level wave functions exhibit the quaternionic analyticity as a generalization of the complex analyticity of the two-dimensional case. Each Landau level contributes one branch of gapless helical Dirac modes to the surface spectra, whose topological properties belong to the Z2 class. The flat Landau levels can be generalized to an arbitrary dimension. Interaction effects and experimental realizations are also studied.

  8. Low copper and high manganese levels in prion protein plaques

    USGS Publications Warehouse

    Johnson, Christopher J.; Gilbert, P.U.P.A.; Abrecth, Mike; Baldwin, Katherine L.; Russell, Robin E.; Pedersen, Joel A.; McKenzie, Debbie

    2013-01-01

    Accumulation of aggregates rich in an abnormally folded form of the prion protein characterize the neurodegeneration caused by transmissible spongiform encephalopathies (TSEs). The molecular triggers of plaque formation and neurodegeneration remain unknown, but analyses of TSE-infected brain homogenates and preparations enriched for abnormal prion protein suggest that reduced levels of copper and increased levels of manganese are associated with disease. The objectives of this study were to: (1) assess copper and manganese levels in healthy and TSE-infected Syrian hamster brain homogenates; (2) determine if the distribution of these metals can be mapped in TSE-infected brain tissue using X-ray photoelectron emission microscopy (X-PEEM) with synchrotron radiation; and (3) use X-PEEM to assess the relative amounts of copper and manganese in prion plaques in situ. In agreement with studies of other TSEs and species, we found reduced brain levels of copper and increased levels of manganese associated with disease in our hamster model. We also found that the in situ levels of these metals in brainstem were sufficient to image by X-PEEM. Using immunolabeled prion plaques in directly adjacent tissue sections to identify regions to image by X-PEEM, we found a statistically significant relationship of copper-manganese dysregulation in prion plaques: copper was depleted whereas manganese was enriched. These data provide evidence for prion plaques altering local transition metal distribution in the TSE-infected central nervous system.

  9. High glucose levels boost the aggressiveness of highly metastatic cholangiocarcinoma cells via O-GlcNAcylation

    PubMed Central

    Phoomak, Chatchai; Vaeteewoottacharn, Kulthida; Silsirivanit, Atit; Saengboonmee, Charupong; Seubwai, Wunchana; Sawanyawisuth, Kanlayanee; Wongkham, Chaisiri; Wongkham, Sopit

    2017-01-01

    Increased glucose utilization is a feature of cancer cells to support cell survival, proliferation, and metastasis. An association between diabetes mellitus and cancer progression was previously demonstrated in cancers including cholangiocarcinoma (CCA). This study was aimed to determine the effects of high glucose on protein O-GlcNAcylation and metastatic potentials of CCA cells. Two pairs each of the parental low metastatic and highly metastatic CCA sublines were cultured in normal (5.6 mM) or high (25 mM) glucose media. The migration and invasion abilities were determined and underlying mechanisms were explored. Results revealed that high glucose promoted migration and invasion of CCA cells that were more pronounced in the highly metastatic sublines. Concomitantly, high glucose increased global O-GlcNAcylated proteins, the expressions of vimentin, hexokinase, glucosamine-fructose-6-phosphate amidotransferase (GFAT) and O-GlcNAc transferase of CCA cells. The glucose level that promoted migration/invasion was shown to be potentiated by the induction of GFAT, O-GlcNAcylation and an increase of O-GlcNAcylated vimentin and vimentin expression. Treatment with a GFAT inhibitor reduced global O-GlcNAcylated proteins, vimentin expression, and alleviated cell migration. Altogether, these results suggested the role of high glucose enhanced CCA metastasis via modulation of O-GlcNAcylation, through the expressions of GFAT and vimentin. PMID:28262738