Sample records for adenoviral sub-unit proteins

  1. Radiolabeled Adenoviral Sub-unit Proteins for Molecular Imaging and Therapeutic Applications in Oncology

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

    Srivastava, S.; Meinken, G.; Springer, K. Awasthi, V.

    2004-10-06

    The objective of this project was to develop and optimize new ligand systems, based on adenoviral vectors (intact adenovirus, adeno-viral fiber protein, and the knob protein), for delivering suitable radionuclides into tumor cells for molecular imaging and combined gene/radionuclide therapy of cancer.

  2. Targeted adenoviral vectors

    NASA Astrophysics Data System (ADS)

    Douglas, Joanne T.

    The practical implementation of gene therapy in the clinical setting mandates gene delivery vehicles, or vectors, capable of efficient gene delivery selectively to the target disease cells. The utility of adenoviral vectors for gene therapy is restricted by their dependence on the native adenoviral primary cellular receptor for cell entry. Therefore, a number of strategies have been developed to allow CAR-independent infection of specific cell types, including the use of bispecific conjugates and genetic modifications to the adenoviral capsid proteins, in particular the fibre protein. These targeted adenoviral vectors have demonstrated efficient gene transfer in vitro , correlating with a therapeutic benefit in preclinical animal models. Such vectors are predicted to possess enhanced efficacy in human clinical studies, although anatomical barriers to their use must be circumvented.

  3. Codon optimization of the adenoviral fiber negatively impacts structural protein expression and viral fitness

    NASA Astrophysics Data System (ADS)

    Villanueva, Eneko; Martí-Solano, Maria; Fillat, Cristina

    2016-06-01

    Codon usage adaptation of lytic viruses to their hosts is determinant for viral fitness. In this work, we analyzed the codon usage of adenoviral proteins by principal component analysis and assessed their codon adaptation to the host. We observed a general clustering of adenoviral proteins according to their function. However, there was a significant variation in the codon preference between the host-interacting fiber protein and the rest of structural late phase proteins, with a non-optimal codon usage of the fiber. To understand the impact of codon bias in the fiber, we optimized the Adenovirus-5 fiber to the codon usage of the hexon structural protein. The optimized fiber displayed increased expression in a non-viral context. However, infection with adenoviruses containing the optimized fiber resulted in decreased expression of the fiber and of wild-type structural proteins. Consequently, this led to a drastic reduction in viral release. The insertion of an exogenous optimized protein as a late gene in the adenovirus with the optimized fiber further interfered with viral fitness. These results highlight the importance of balancing codon usage in viral proteins to adequately exploit cellular resources for efficient infection and open new opportunities to regulate viral fitness for virotherapy and vaccine development.

  4. Avidin-Based Targeting and Purification of a Protein IX-Modified, Metabolically Biotinylated Adenoviral Vector

    PubMed Central

    Campos, Samuel K.; Parrott, M. Brandon; Barry, Michael A.

    2014-01-01

    While genetic modification of adenoviral vectors can produce vectors with modified tropism, incorporation of targeting peptides/proteins into the structural context of the virion can also result in destruction of ligand targeting or virion integrity. To combat this problem, we have developed a versatile targeting system using metabolically biotinylated adenoviral vectors bearing biotinylated fiber proteins. These vectors have been demonstrated to be useful as a platform for avidin-based ligand screening and vector targeting by conjugating biotinylated ligands to the virus using high-affinity tetrameric avidin (Kd = 10−15 M). The biotinylated vector could also be purified by biotin-reversible binding on monomeric avidin (Kd = 10−7 M). In this report, a second metabolically biotinylated adenovirus vector, Ad-IX-BAP, has been engineered by fusing a biotin acceptor peptide (BAP) to the C-terminus of the adenovirus pIX protein. This biotinylated vector displays twice as many biotins and was markedly superior for single-step affinity purification on monomeric avidin resin. However, unlike the fiber-biotinylated vector, Ad-IX-BAP failed to retarget to cells with biotinylated antibodies including anti-CD71 against the transferrin receptor. In contrast, Ad-IX-BAP was retargeted if transferrin, the cognate ligand for CD71, was used as a ligand rather than the anti-CD71. This work demonstrates the utility of metabolic biotinylation as a molecular screening tool to assess the utility of different viral capsid proteins for ligand display and the biology and compatibility of different ligands and receptors for vector targeting applications. These results also demonstrate the utility of the pIX-biotinylated vector as a platform for gentle single-step affinity purification of adenoviral vectors. PMID:15194061

  5. Bacterial RecA Protein Promotes Adenoviral Recombination during In Vitro Infection

    PubMed Central

    Lee, Jeong Yoon; Lee, Ji Sun; Materne, Emma C.; Rajala, Rahul; Ismail, Ashrafali M.; Seto, Donald; Dyer, David W.

    2018-01-01

    presence of bacterial RecA protein facilitated homologous recombination between viruses. Genetic recombination led to evolution of an important external feature on the adenoviral capsid, namely, the penton base protein hypervariable loop 2, which contains the arginine-glycine-aspartic acid motif critical to viral internalization. We speculate that free Rec proteins present in gastrointestinal secretions upon bacterial cell death facilitate the evolution of human adenoviruses through homologous recombination, an example of viral commensalism and the complexity of virus-host interactions, including regional microbiota. PMID:29925671

  6. Germline incorporation of a replication-defective adenoviral vector in mice does not alter immune responses to adenoviral vectors.

    PubMed

    Camargo, F D; Huey-Louie, D A; Finn, A V; Sassani, A B; Cozen, A E; Moriwaki, H; Schneider, D B; Agah, R; Dichek, D A

    2000-11-01

    The utility of adenoviral vectors is limited by immune responses to adenoviral antigens. We sought to develop immune-competent mice in which the immune response to adenoviral antigens was selectively absent. To do so, we generated mice that were transgenic for a replication-defective vector. Adenoviral antigens might be seen as self-antigens by these mice, and the mice could exhibit immunologic tolerance after postnatal exposure to adenoviral vectors. In addition, characterization of these mice could reveal potential consequences of germline transmission of an adenoviral vector, as might occur in a gene therapy trial. Injection of a "null" (not containing a transgene) E1, E3-deleted vector genome into mouse zygotes yielded five founders that were capable of transmitting the vector genome. Among offspring of these mice, transgenic pups were significantly underrepresented: 108 of 255 pups (42%) were transgenic (P<0.02 versus expected frequency of 50%). Postnatal transgenic mice, however, had no apparent abnormalities. Persistence of an adenoviral vector after intravenous injection was equivalent in livers of transgenic mice and their nontransgenic littermates. Transgenic and nontransgenic mice also had equivalent humoral and cellular immune responses to adenoviral vector injection. Mice that are transgenic for an E1, E3-deleted adenoviral genome can be easily generated; however, they are not tolerant of adenovirus. Moreover, germline transmission of an adenoviral vector genome does not prevent generation of a robust immune response after exposure to adenoviral antigens.

  7. High-capacity 'gutless' adenoviral vectors.

    PubMed

    Kochanek, S; Schiedner, G; Volpers, C

    2001-10-01

    Adenoviral vectors are promising gene transfer vehicles for different gene therapy applications. High-capacity adenoviral (HC-Ad) vectors address some of the problems that have been observed with replication-defective, E1-deleted first-generation adenoviral vectors: toxicity and immunogenicity due to viral gene expression and 7 to 8 kb capacity limit for the transport of therapeutic DNA. This review summarizes HC-Ad vector-related publications from the past 18 months that are mainly concerned with vector design/production and in vivo applications in different murine models.

  8. MicroRNA Silencing Improves the Tumor Specificity of Adenoviral Transgene Expression

    PubMed Central

    Card, Paul B.; Hogg, Richard T.; del Alcazar, Carlos Gil

    2012-01-01

    Adenoviral technology has been thoroughly evaluated for delivering genetic material to tumor tissue and the surrounding microenvironment. Almost any gene can be cloned into an adenovirus (Ad) vector, which when combined with strong, constitutively active promoters permit up to a million-fold amplification of the transgene in a single adenoviral particle, thus facilitating their use in cancer therapy and imaging. However, widespread infection of the liver and other non-targeted tissues by Ad vectors is a substantial problem that often results in significant liver inflammation and hepatotoxicity at doses required to achieve efficient tumor transduction. miR-122 is a highly expressed liver-specific microRNA that provides a unique opportunity for down-regulating adenoviral transgene expression in liver tissue. The binding of endogenous miRNAs to complementary miRNA targeting elements (miRTs) incorporated into the 3′ untranslated region of adenoviral transgenes interferes with message stability and/or protein translation, and miRT elements against miR-122 (miRT-122) can selectively reduce adenoviral transgene expression in the liver. Previous studies using miR-122-based regulation, with and without other types of transcriptional targeting, have yielded promising preliminary results. However, investigations to date evaluating miRT-122 elements for improving tumor specificity have used either non-tumor bearing animals or direct intratumoral injection as the mode of delivery. In the present study, we confirmed the ability of miRT-122 sequences to selectively down-regulate adenoviral luciferase expression in the liver in vitro and in vivo, and show that this strategy can improve tumor specific transgene expression in a HT1080 human fibrosarcoma model. Rapid growth and the inefficient flow of blood through tumor neovasculature often results in profound hypoxia, which provides additional opportunities for targeting solid tumors and their microenvironment using vectors

  9. Modification of liposomal concentration in liposome/adenoviral complexes allows significant protection of adenoviral vectors from neutralising antibody, in vitro.

    PubMed

    Steel, Jason C; Cavanagh, Heather M A; Burton, Mark A; Dingwall, Daniel J; Kalle, Wouter H J

    2005-06-01

    Adenoviral vectors have been commonly used in gene therapy protocols, however the success of their use is often limited by the induction of host immunity to the vector. Following exposure to the adenoviral vector, adenoviral-specific neutralising antibodies are produced which limits further administration. This study examines the efficacy of complexing liposomes to adenovirus for the protection of the adenovirus from neutralising antibodies in an in vitro setting. Dimethyldioctadecylammonium bromide (DDAB)-dioleoyl-l-phosphatidylethanolamine (DOPE) liposomes were bound at varying concentrations to adenovirus to form AL complexes and tested these complexes' ability to prevent adenoviral neutralisation. It is shown that by increasing the concentration of liposomes in the adenoviral-liposome (AL) complexes we can increase the level of immuno-shielding afforded the adenovirus. It is also shown that the increase in liposomal concentration may lead to drawbacks such as increased cytotoxicity and reductions in expression levels.

  10. In vivo, cardiac-specific knockdown of target protein, Malic Enzyme-1, in rat via adenoviral delivery of DNA for non-native miRNA

    PubMed Central

    O'Donnell, J. Michael; Kalichira, Asha; Bi, Jian; Lewandowski, E. Douglas

    2013-01-01

    This study examines the feasibility of using the adenoviral delivery of DNA for a non-native microRNA to suppress expression of a target protein (cytosolic NADP+-dependent malic-enzyme 1, ME1) in whole heart in vivo, via an isolated-heart coronary perfusion approach. Complementary DNA constructs for ME1 microRNA were inserted into adenoviral vectors. Viral gene transfer to neonatal rat cardiomyocytes yielded 65% suppression of ME1 protein. This viral package was delivered to rat hearts in vivo (Adv.miR_ME1, 1013 vp/ml PBS) via coronary perfusion, using a cardiac-specific isolation technique. ME1 mRNA was reduced by 73% at 2-6 days post-surgery in heart receiving the Adv.miR_ME1. Importantly, ME1 protein was reduced by 66% (p<0.0002) at 5-6 days relative to sham-operated control hearts. Non-target protein expression for GAPDH, calsequestrin, and mitochondrial malic enzyme, ME3, were all unchanged. The non-target isoform, ME2, was unchanged at 2-5 days and reduced at day 6. This new approach demonstrates for the first time significant and acute silencing of target RNA translation and protein content in whole heart, in vivo, via non-native microRNA expression. PMID:22974418

  11. Interleukin-Encoding Adenoviral Vectors as Genetic Adjuvant for Vaccination against Retroviral Infection

    PubMed Central

    Ohs, Inga; Windmann, Sonja; Wildner, Oliver; Dittmer, Ulf; Bayer, Wibke

    2013-01-01

    Interleukins (IL) are cytokines with stimulatory and modulatory functions in the immune system. In this study, we have chosen interleukins which are involved in the enhancement of TH2 responses and B cell functions to analyze their potential to improve a prophylactic adenovirus-based anti-retroviral vaccine with regard to antibody and virus-specific CD4+ T cell responses. Mice were vaccinated with an adenoviral vector which encodes and displays the Friend Virus (FV) surface envelope protein gp70 (Ad.pIXgp70) in combination with adenoviral vectors encoding the interleukins IL4, IL5, IL6, IL7 or IL23. Co-application of Ad.pIXgp70 with Ad.IL5, Ad.IL6 or Ad.IL23 resulted in improved protection with high control over FV-induced splenomegaly and reduced viral loads. Mice co-immunized with adenoviral vectors encoding IL5 or IL23 showed increased neutralizing antibody responses while mice co-immunized with Ad.IL6 or Ad.IL23 showed improved FV-specific CD4+ T cell responses compared to mice immunized with Ad.pIXgp70 alone. We show that the co-application of adenoviral vectors encoding specific interleukins is suitable to improve the vaccination efficacy of an anti-retroviral vaccine. Improved protection correlated with improved CD4+ T cell responses and especially with higher neutralizing antibody titers. The co-application of selected interleukin-encoding adenoviral vectors is a valuable tool for vaccination with regard to enhancement of antibody mediated immunity. PMID:24349306

  12. Optimizing cardiovascular gene therapy: increased vascular gene transfer with modified adenoviral vectors.

    PubMed

    Kibbe, M R; Murdock, A; Wickham, T; Lizonova, A; Kovesdi, I; Nie, S; Shears, L; Billiar, T R; Tzeng, E

    2000-02-01

    Adenovirus is widely used as a vector for gene transfer to the vasculature. However, the efficiency of these vectors can be limited by ineffective viral-target cell interactions. Viral attachment, which largely determines adenoviral tropism, is mediated through binding of the adenoviral fiber coat protein to the Coxsackievirus and adenovirus receptor, while internalization follows binding of the adenoviral RGD motif to alpha(v)-integrin receptors. Modifications of the fiber coat protein sequence have been successful for targeting the adenovirus to more prevalent receptors in the vasculature, including heparan sulfate-containing receptors and alpha(v)-integrin receptors. Modified adenoviral vectors targeted to receptors more prevalent in the vasculature result in an increased transfer efficiency of the virus in vitro and in vivo even in the presence of clinically relevant doses of heparin. We tested 2 modified E1- and E3-deleted Ad5 type adenoviral vectors containing the beta-galactosidase gene. AdZ.F(pK7) contains multiple positively charged lysines in the fiber coat protein that target the adenovirus to heparan sulfate receptors, while AdZ.F(RGD) contains an RGD integrin-binding sequence in the fiber coat protein that allows binding to alpha(v)-integrin receptors. The gene transfer efficiency of these modified viruses was compared in rat aortic smooth muscle cells in vitro and in an in vivo porcine model of balloon-induced arterial injury. Because of the use of heparin during most vascular surgical procedures and the concern that heparin might interfere with the binding of AdZ.F(pK7) to heparan sulfate receptors, the effect of heparin on the in vitro and in vivo transfer efficiency of these 2 modified adenoviruses was evaluated. In vitro infection of rat aortic smooth muscle cells with AdZ.F(pK7) and AdZ.F(RGD) resulted in significantly higher levels of beta-galactosidase expression compared with the unmodified adenovirus (mean +/- SEM, 1766.3 +/- 89.1 and 44

  13. Neonatal adenoviral infection: a seventeen year experience and review of the literature.

    PubMed

    Ronchi, Andrea; Doern, Christopher; Brock, Evangeline; Pugni, Lorenza; Sánchez, Pablo J

    2014-03-01

    To describe the clinical manifestations and short-term outcomes of adenoviral infections in neonates and review all published cases to better determine impact and treatment outcomes. Retrospective cohort study of all neonates hospitalized at Children's Medical Center (CMC) and Parkland Memorial Hospital (PMH), Dallas, TX with laboratory-confirmed adenoviral infection from January 1,1995-December 31, 2012. Neonates were identified by review of the CMC Virology Laboratory's prospective database of all positive adenovirus tests performed in the inpatient and ambulatory settings, and at PMH, of a prospective neonatal database that included all neonatal intensive care unit admissions. Patients also were identified by discharge International Classification of Disease, 9th edition codes for adenoviral infection. The medical records were reviewed, and a review of the English literature was performed. During 17 years, 26 neonates had adenoviral infection (25, CMC; 1, PMH). The principle reasons for hospitalization were respiratory signs (88%) and temperature instability (65%). Five (19%) had disseminated disease and 4 (80%) of these infants died. Ribavirin or cidofovir treatment, as well as immune globulin intravenous, did not improve outcomes except in 1 neonate. Literature review (n = 72) combined with our data found that disseminated infection was associated with death (68% vs 21% with localized infection, P < .001). In addition, neonates <14 days of age were more likely to have disseminated disease (44% vs 12%, P = .004) and death (48% vs 8%; P < .001). Adenoviral infection in hospitalized neonates was associated with severe morbidity and mortality, especially when infection was disseminated and involved the respiratory tract. Development of new therapeutic strategies is needed. Copyright © 2014 Mosby, Inc. All rights reserved.

  14. Gene Transfer into Rat Brain Using Adenoviral Vectors

    PubMed Central

    Puntel, Mariana; Kroeger, Kurt M.; Sanderson, Nicholas S.R.; Thomas, Clare E.; Castro, Maria G.; Lowenstein, Pedro R.

    2010-01-01

    Viral vector–mediated gene delivery is an attractive procedure for introducing genes into the brain, both for purposes of basic neuroscience research and to develop gene therapy for neurological diseases. Replication-defective adenoviruses possess many features which make them ideal vectors for this purpose—efficiently transducing terminally differentiated cells such as neurons and glial cells, resulting in high levels of transgene expression in vivo. Also, in the absence of anti-adenovirus immunity, these vectors can sustain very long-term transgene expression within the brain parenchyma. This unit provides protocols for the stereotactic injection of adenoviral vectors into the brain, followed by protocols to detect transgene expression or infiltrates of immune cells by immunocytochemistry or immunofluorescence. ELISPOT and neutralizing antibody assay methodologies are provided to quantitate the levels of cellular and humoral immune responses against adenoviruses. Quantitation of adenoviral vector genomes within the rat brain using qPCR is also described. Curr. Protoc. Neurosci. 50:4.24.1–4.24.49. © 2010 by John Wiley & Sons, Inc. PMID:20066657

  15. ROLE OF GENETIC SUSCEPTIBILITY TO LATENT ADENOVIRAL INFECTION AND DECREASED LUNG FUNCTION

    PubMed Central

    Kasuga, Ikuma; Hogg, James C.; Paré, Peter D.; Hayashi, Shizu; Sedgwick, Edward G.; Ruan, Jian; Wallace, Alison M.; He, Jian-Qing; Zhang, Xiaozhu; Sandford, Andrew J.

    2009-01-01

    Background Latent adenoviral infection may amplify cigarette smoke-induced lung inflammation and therefore play an important role in the development of chronic obstructive pulmonary disease (COPD). Adenoviruses can evade the human immune response via their 19-kDa protein (19K) which delays the expression of class I human leukocyte antigen (HLA) proteins. The 19K protein shows higher affinity to HLA-B7 and A2 compared with HLA-A1 and A3. The receptor for adenovirus (CXADR) and integrin β5 (ITGB5) are host factors which might affect adenovirus infection. Therefore, we investigated the contribution of HLA, CXADR, and ITGB5 genetic variants to the presence of the E1A gene and to level of lung function. Methods Study subjects were assayed for HLA-B7, A1, A2 and A3 by PCR-based assays using allele-specific primers. Polymorphisms of the CXADR and ITGB5 genes were genotyped by PCR-based restriction fragment length polymorphism assays. Detection of adenoviral E1A gene was performed by a real-time PCR TaqMan assay. Results E1A positive individuals have a lower FEV1 compared with E1A negative individuals. However, there was no significant difference in E1A positivity rate between the high (HLA-B7 and A2) and low (HLA-A1 and A3) 19K affinity groups. There was also no significant difference in FEV1 level between each affinity group. There was no significant difference in E1A positivity rate or lung function among the CXADR and ITGB5 genotypes. Conclusions Genetic variants in HLA, CXADR and ITGB5 do not influence latent adenoviral infections and are not associated with COPD. PMID:19502044

  16. Vaccination with an adenoviral vector that encodes and displays a retroviral antigen induces improved neutralizing antibody and CD4+ T-cell responses and confers enhanced protection.

    PubMed

    Bayer, Wibke; Tenbusch, Matthias; Lietz, Ruth; Johrden, Lena; Schimmer, Simone; Uberla, Klaus; Dittmer, Ulf; Wildner, Oliver

    2010-02-01

    We present a new type of adenoviral vector that both encodes and displays a vaccine antigen on the capsid, thus combining in itself gene-based and protein vaccination; this vector resulted in an improved vaccination outcome in the Friend virus (FV) model. For presentation of the envelope protein gp70 of Friend murine leukemia virus on the adenoviral capsid, gp70 was fused to the adenovirus capsid protein IX. When compared to vaccination with conventional FV Env- and Gag-encoding adenoviral vectors, vaccination with the adenoviral vector that encodes and displays pIX-gp70 combined with an FV Gag-encoding vector resulted in significantly improved protection against systemic FV challenge infection, with highly controlled viral loads in plasma and spleen. This improved protection correlated with improved neutralizing antibody titers and stronger CD4(+) T-cell responses. Using a vector that displays gp70 without encoding it, we found that while the antigen display on the capsid alone was sufficient to induce high levels of binding antibodies, in vivo expression was necessary for the induction of neutralizing antibodies. This new type of adenovirus-based vaccine could be a valuable tool for vaccination.

  17. Anti-adenoviral effect of anti-HIV agents in vitro in serotypes inducing keratoconjunctivitis.

    PubMed

    Uchio, Eiichi; Fuchigami, Aki; Kadonosono, Kazuaki; Hayashi, Akio; Ishiko, Hiroaki; Aoki, Koki; Ohno, Shigeaki

    2007-09-01

    Around one million people are affected by adenoviral keratoconjunctivitis a year in Japan, and it is recognized as one of the major pathogens of ophthalmological nosocomial infection worldwide. Although cidofovir can be used systemically for immunocompromised patients with disseminated adenoviral infection, no specific anti-adenoviral agent has been established for the treatment of adenoviral infection. We evaluated the anti-adenoviral effect of anti-HIV (human immunodeficiency virus) agents in this study. Five anti-HIV agents (zalcitabine, stavudine, nevirapine, indinavir and amprenavir) were subjected to in vitro evaluation. A549 cells were used for viral cell culture, and adenovirus serotypes 3, 4, 8, 19 and 37 were used. After calculating CC(50) (50% cytotoxic concentration) of each agent by MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) method, we cultured adenovirus with the agents for seven days and quantitatively measured extracted adenoviral DNA by real-time PCR. Among the anti-HIV drugs, zalcitabine and stavudine, both nucleoside reverse transcriptase inhibitors, showed significant anti-adenoviral activity. In contrast, nevirapine, a non-nucleoside reverse transcriptase inhibitor, and indinavir and amprenavir, which are both protease inhibitors, were ineffective against adenovirus. These results indicate that zalcitabine and stavudine are possible candidates for the local and systemic treatment of adenoviral infection, and the anti-adenoviral effect might depend on the pharmacological properties of anti-HIV agents. The chemical properties on the clinical safety for systemic and local application need to be determined in order to for these drugs to be accepted for the treatment of adenovirus in clinical settings.

  18. Isolation and characterization of anti-adenoviral secondary metabolites from marine actinobacteria.

    PubMed

    Strand, Mårten; Carlsson, Marcus; Uvell, Hanna; Islam, Koushikul; Edlund, Karin; Cullman, Inger; Altermark, Björn; Mei, Ya-Fang; Elofsson, Mikael; Willassen, Nils-Peder; Wadell, Göran; Almqvist, Fredrik

    2014-01-28

    Adenovirus infections in immunocompromised patients are associated with high mortality rates. Currently, there are no effective anti-adenoviral therapies available. It is well known that actinobacteria can produce secondary metabolites that are attractive in drug discovery due to their structural diversity and their evolved interaction with biomolecules. Here, we have established an extract library derived from actinobacteria isolated from Vestfjorden, Norway, and performed a screening campaign to discover anti-adenoviral compounds. One extract with anti-adenoviral activity was found to contain a diastereomeric 1:1 mixture of the butenolide secondary alcohols 1a and 1b. By further cultivation and analysis, we could isolate 1a and 1b in different diastereomeric ratio. In addition, three more anti-adenoviral butenolides 2, 3 and 4 with differences in their side-chains were isolated. In this study, the anti-adenoviral activity of these compounds was characterized and substantial differences in the cytotoxic potential between the butenolide analogs were observed. The most potent butenolide analog 3 displayed an EC50 value of 91 μM and no prominent cytotoxicity at 2 mM. Furthermore, we propose a biosynthetic pathway for these compounds based on their relative time of appearance and structure.

  19. Regulation of human adenovirus alternative RNA splicing by the adenoviral L4-33K and L4-22K proteins.

    PubMed

    Biasiotto, Roberta; Akusjärvi, Göran

    2015-01-28

    Adenovirus makes extensive use of alternative RNA splicing to produce a complex set of spliced viral mRNAs. Studies aimed at characterizing the interactions between the virus and the host cell RNA splicing machinery have identified three viral proteins of special significance for the control of late viral gene expression: L4-33K, L4-22K, and E4-ORF4. L4-33K is a viral alternative RNA splicing factor that controls L1 alternative splicing via an interaction with the cellular protein kinases Protein Kinase A (PKA) and DNA-dependent protein kinase (DNA-PK). L4-22K is a viral transcription factor that also has been implicated in the splicing of a subset of late viral mRNAs. E4-ORF4 is a viral protein that binds the cellular protein phosphatase IIA (PP2A) and controls Serine/Arginine (SR)-rich protein activity by inducing SR protein dephosphorylation. The L4-33K, and most likely also the L4-22K protein, are highly phosphorylated in vivo. Here we will review the function of these viral proteins in the post-transcriptional control of adenoviral gene expression and further discuss the significance of potential protein kinases phosphorylating the L4-33K and/or L4-22K proteins.

  20. Adenoviral bcl-2 transfer improves survival and early graft function after ischemia and reperfusion in rat liver transplantation.

    PubMed

    Rentsch, Markus; Kienle, Klaus; Mueller, Thomas; Vogel, Mandy; Jauch, Karl Walter; Püllmann, Kerstin; Obed, Aiman; Schlitt, Hans J; Beham, Alexander

    2005-11-27

    Primary graft dysfunction due to ischemia and reperfusion injury represents a major problem in liver transplantation. The related cell stress may induce apoptosis, which can be suppressed by bcl-2. The purpose of the study was to investigate the effect of adenoviral bcl-2 gene transfer on early graft function and survival in rat liver transplantation. An adenoviral construct that transfers bcl-2 under the control of a tetracycline inducible promoter was generated (advTetOn bcl-2) and used with a second adenovirus that transfers the repressor protein (advCMV Rep). Forty-eight hours before explantation, donor rats were treated with advTetOn bcl-2/ advCMV Rep (n=7) and doxycyclin, with the control adenoviral construct advCMV GFP (n=8) or with doxycyclin alone (n=8). Liver transplantation was performed following 16 hours of cold storage (UW). Bcl-2 expression and intrahepatic apoptosis was assessed. Bile flow was monitored 90 min posttransplantation. The endpoint for survival was 7 days. Bcl-2 was expressed in hepatocytes and sinusoidal lining cells. This was associated with a significant reduction of apoptotic sinusoidal lining cells and hepatocytes after 24 hours and 7 days. Bile production was significantly higher following bcl-2 pretreatment. Furthermore, bcl-2 transfer resulted in significantly improved survival (100% vs. 50% both control groups). Adenoviral bcl-2 transfer results in protein expression in hepatocytes and sinusoidal lining cells resulting in early graft function and survival enhancement after prolonged ischemia and reperfusion injury. The inhibition of apoptosis in the context of liver transplantation might be a reasonable approach in the treatment of graft dysfunction.

  1. Rapid construction of capsid-modified adenoviral vectors through bacteriophage lambda Red recombination.

    PubMed

    Campos, Samuel K; Barry, Michael A

    2004-11-01

    There are extensive efforts to develop cell-targeting adenoviral vectors for gene therapy wherein endogenous cell-binding ligands are ablated and exogenous ligands are introduced by genetic means. Although current approaches can genetically manipulate the capsid genes of adenoviral vectors, these approaches can be time-consuming and require multiple steps to produce a modified viral genome. We present here the use of the bacteriophage lambda Red recombination system as a valuable tool for the easy and rapid construction of capsid-modified adenoviral genomes.

  2. Adenoviral Gene Delivery to Primary Human Cutaneous Cells and Burn Wounds

    PubMed Central

    Hirsch, Tobias; von Peter, Sebastian; Dubin, Grzegorz; Mittler, Dominik; Jacobsen, Frank; Lehnhardt, Markus; Eriksson, Elof; Steinau, Hans-Ulrich; Steinstraesser, Lars

    2006-01-01

    The adenoviral transfer of therapeutic genes into epidermal and dermal cells is an interesting approach to treat skin diseases and to promote wound healing. The aim of this study was to assess the in vitro and in vivo transfection efficacy in skin and burn wounds after adenoviral gene delivery. Primary keratinocytes (HKC), fibroblasts (HFB), and HaCaT cells were transfected using different concentrations of an adenoviral construct (eGFP). Transfection efficiency and cytotoxicity was determined up to 30 days. Expression was quantified by FACS analysis and fluorimeter. Cytotoxicity was measured using the trypan blue exclusion method. 45 male Sprague Dawley rats received 2 × 108 pfu of Ad5-CMV-LacZ or carrier control intradermally into either superficial partial thickness scald burn or unburned skin. Animals were euthanized after 48 h, 7 or 14 days posttreatment. Transgene expression was assessed using immunohistochemistry and bioluminescent assays. The highest transfection rate was observed 48 h posttransfection: 79% for HKC, 70% for HFB, and 48% for HaCaT. The eGFP expression was detectable in all groups over 30 days (P > 0.05). Cytotoxic effects of the adenoviral vector were observed for HFB after 10 days and HaCaT after 30 days. Reporter gene expression in vivo was significantly higher in burned skin compared with unburned skin (P = 0,004). Gene expression decreases from 2 to 7 days with no significant expression after 14 days. This study demonstrates that effective adenoviral-mediated gene transfer of epidermal primary cells and cell-lines is feasible. Ex vivo gene transfer in epithelial cells might have promise for the use in severely burned patients who receive autologous keratinocyte sheets. Transient cutaneous gene delivery in burn wounds using adenoviral vectors causes significant concentrations in the wound tissue for at least 1 week. Based on these findings, we hypothesize that transient cutaneous adenoviral gene delivery of wound healing promoting

  3. Adenoviral gene delivery to primary human cutaneous cells and burn wounds.

    PubMed

    Hirsch, Tobias; von Peter, Sebastian; Dubin, Grzegorz; Mittler, Dominik; Jacobsen, Frank; Lehnhardt, Markus; Eriksson, Elof; Steinau, Hans-Ulrich; Steinstraesser, Lars

    2006-01-01

    The adenoviral transfer of therapeutic genes into epidermal and dermal cells is an interesting approach to treat skin diseases and to promote wound healing. The aim of this study was to assess the in vitro and in vivo transfection efficacy in skin and burn wounds after adenoviral gene delivery. Primary keratinocytes (HKC), fibroblasts (HFB), and HaCaT cells were transfected using different concentrations of an adenoviral construct (eGFP). Transfection efficiency and cytotoxicity was determined up to 30 days. Expression was quantified by FACS analysis and fluorimeter. Cytotoxicity was measured using the trypan blue exclusion method. 45 male Sprague Dawley rats received 2x10(8) pfu of Ad5-CMV-LacZ or carrier control intradermally into either superficial partial thickness scald burn or unburned skin. Animals were euthanized after 48 h, 7 or 14 days posttreatment. Transgene expression was assessed using immunohistochemistry and bioluminescent assays. The highest transfection rate was observed 48 h posttransfection: 79% for HKC, 70% for HFB, and 48% for HaCaT. The eGFP expression was detectable in all groups over 30 days (P>0.05). Cytotoxic effects of the adenoviral vector were observed for HFB after 10 days and HaCaT after 30 days. Reporter gene expression in vivo was significantly higher in burned skin compared with unburned skin (P=0,004). Gene expression decreases from 2 to 7 days with no significant expression after 14 days. This study demonstrates that effective adenoviral-mediated gene transfer of epidermal primary cells and cell-lines is feasible. Ex vivo gene transfer in epithelial cells might have promise for the use in severely burned patients who receive autologous keratinocyte sheets. Transient cutaneous gene delivery in burn wounds using adenoviral vectors causes significant concentrations in the wound tissue for at least 1 week. Based on these findings, we hypothesize that transient cutaneous adenoviral gene delivery of wound healing promoting factors has

  4. Heterologous prime-boost regimens with a recombinant chimpanzee adenoviral vector and adjuvanted F4 protein elicit polyfunctional HIV-1-specific T-Cell responses in macaques.

    PubMed

    Lorin, Clarisse; Vanloubbeeck, Yannick; Baudart, Sébastien; Ska, Michaël; Bayat, Babak; Brauers, Geoffroy; Clarinval, Géraldine; Donner, Marie-Noëlle; Marchand, Martine; Koutsoukos, Marguerite; Mettens, Pascal; Cohen, Joe; Voss, Gerald

    2015-01-01

    HIV-1-specific CD4+ and CD8+ T lymphocytes are important for HIV-1 replication control. F4/AS01 consists of F4 recombinant fusion protein (containing clade B Gag/p24, Pol/RT, Nef and Gag/p17) formulated in AS01 Adjuvant System, and was shown to induce F4-specific polyfunctional CD4+ T-cell responses in humans. While replication-incompetent recombinant HIV-1/SIV antigen-expressing human adenoviral vectors can elicit high-frequency antigen-specific CD8+ T-cell responses, their use is hampered by widespread pre-existing immunity to human serotypes. Non-human adenovirus serotypes associated with lower prevalence may offer an alternative strategy. We evaluated the immunogenicity of AdC7-GRN ('A'), a recombinant chimpanzee adenovirus type 7 vector expressing clade B Gag, RT and Nef, and F4/AS01 ('P'), when delivered intramuscularly in homologous (PP or AA) and heterologous (AAPP or PPAA) prime-boost regimens, in macaques and mice. Vaccine-induced HIV-1-antigen-specific T cells in peripheral blood (macaques), liver, spleen, and intestinal and genital mucosa (mice) were characterized by intracellular cytokine staining. Vaccine-specific IgG antibodies (macaques) were detected using ELISA. In macaques, only the heterologous prime-boost regimens induced polyfunctional, persistent and balanced CD4+ and CD8+ T-cell responses specific to each HIV-1 vaccine antigen. AdC7-GRN priming increased the polyfunctionality of F4/AS01-induced CD4+ T cells. Approximately 50% of AdC7-GRN-induced memory CD8+ T cells exhibited an effector-memory phenotype. HIV-1-specific antibodies were detected with each regimen. In mice, antigen-specific CD4+ and CD8+ T-cell responses were detected in the mucosal and systemic anatomical compartments assessed. When administered in heterologous prime-boost regimens, AdC7-GRN and F4/AS01 candidate vaccines acted complementarily in inducing potent and persistent peripheral blood HIV-1-specific CD4+ and CD8+ T-cell responses and antibodies in macaques. Besides

  5. Heterologous Prime-Boost Regimens with a Recombinant Chimpanzee Adenoviral Vector and Adjuvanted F4 Protein Elicit Polyfunctional HIV-1-Specific T-Cell Responses in Macaques

    PubMed Central

    Lorin, Clarisse; Vanloubbeeck, Yannick; Baudart, Sébastien; Ska, Michaël; Bayat, Babak; Brauers, Geoffroy; Clarinval, Géraldine; Donner, Marie-Noëlle; Marchand, Martine; Koutsoukos, Marguerite; Mettens, Pascal; Cohen, Joe; Voss, Gerald

    2015-01-01

    HIV-1-specific CD4+ and CD8+ T lymphocytes are important for HIV-1 replication control. F4/AS01 consists of F4 recombinant fusion protein (containing clade B Gag/p24, Pol/RT, Nef and Gag/p17) formulated in AS01 Adjuvant System, and was shown to induce F4-specific polyfunctional CD4+ T-cell responses in humans. While replication-incompetent recombinant HIV-1/SIV antigen-expressing human adenoviral vectors can elicit high-frequency antigen-specific CD8+ T-cell responses, their use is hampered by widespread pre-existing immunity to human serotypes. Non-human adenovirus serotypes associated with lower prevalence may offer an alternative strategy. We evaluated the immunogenicity of AdC7-GRN (‘A’), a recombinant chimpanzee adenovirus type 7 vector expressing clade B Gag, RT and Nef, and F4/AS01 (‘P’), when delivered intramuscularly in homologous (PP or AA) and heterologous (AAPP or PPAA) prime-boost regimens, in macaques and mice. Vaccine-induced HIV-1-antigen-specific T cells in peripheral blood (macaques), liver, spleen, and intestinal and genital mucosa (mice) were characterized by intracellular cytokine staining. Vaccine-specific IgG antibodies (macaques) were detected using ELISA. In macaques, only the heterologous prime-boost regimens induced polyfunctional, persistent and balanced CD4+ and CD8+ T-cell responses specific to each HIV-1 vaccine antigen. AdC7-GRN priming increased the polyfunctionality of F4/AS01-induced CD4+ T cells. Approximately 50% of AdC7-GRN-induced memory CD8+ T cells exhibited an effector-memory phenotype. HIV-1-specific antibodies were detected with each regimen. In mice, antigen-specific CD4+ and CD8+ T-cell responses were detected in the mucosal and systemic anatomical compartments assessed. When administered in heterologous prime-boost regimens, AdC7-GRN and F4/AS01 candidate vaccines acted complementarily in inducing potent and persistent peripheral blood HIV-1-specific CD4+ and CD8+ T-cell responses and antibodies in macaques

  6. Adenoviral transduction supports matrix expression of alginate cultured articular chondrocytes.

    PubMed

    Pohle, D; Kasch, R; Herlyn, P; Bader, R; Mittlmeier, T; Pützer, B M; Müller-Hilke, B

    2012-09-01

    The present study examines the effects of adenoviral (Ad) transduction of human primary chondrocyte on transgene expression and matrix production. Primary chondrocytes were isolated from healthy articular cartilage and from cartilage with mild osteoarthritis (OA), transduced with an Ad vector and either immediately cultured in alginate or expanded in monolayer before alginate culture. Proteoglycan production was measured using dimethylmethylene blue (DMMB) assay and matrix gene expression was quantified by real-time PCR. Viral infection of primary chondrocytes results in a stable long time transgene expression for up to 13 weeks. Ad transduction does not significantly alter gene expression and matrix production if chondrocytes are immediately embedded in alginate. However, if expanded prior to three dimension (3D) culture in alginate, chondrocytes produce not only more proteoglycans compared to non-transduced controls, but also display an increased anabolic and decreased catabolic activity compared to non-transduced controls. We therefore suggest that successful autologous chondrocyte transplantation (ACT) should combine adenoviral transduction of primary chondrocytes with expansion in monolayer followed by 3D culture. Future studies will be needed to investigate whether the subsequent matrix production can be further improved by using Ad vectors bearing genes encoding matrix proteins. Copyright © 2012 Wiley Periodicals, Inc.

  7. Purification of adenoviral vectors by combined anion exchange and gel filtration chromatography.

    PubMed

    Eglon, Marc N; Duffy, Aoife M; O'Brien, Timothy; Strappe, Padraig M

    2009-11-01

    Adenoviral vectors are used extensively in human gene therapy trials and in vaccine development. Large-scale GMP production requires a downstream purification process, and liquid chromatography is emerging as the most powerful mode of purification, enabling the production of vectors at a clinically relevant scale and quality. The present study describes the development of a two-step high-performance liquid chromatography (HPLC) process combining anion exchange (AIEX) and gel filtration (GF) in comparison with the caesium chloride density gradient method. HEK-293 cells were cultured in ten-layer CellStacks() and infected with 10 pfu/cell of adenoviral vector expressing green fluorescent protein (Ad5-GFP). Cell-bound virus was harvested and benzonase added to digest DNA, crude lysate was clarified by centrifugation and filtration prior to HPLC. Chromatography fractions were added to HEK-293 cells and GFP expression measured using a fluorescent plate reader. Using AIEX then GF resulted in an adenoviral vector with purity comparable to Ad5-GFP purified by CsCl, whereas the reverse process (GF-AIEX) showed a reduced purity by electrophoresis and required further buffer exchange of the product. The optimal process (AIEX-GF) resulted in a vector yield of 2.3 x 10(7) pfu/cm(2) of cell culture harvested compared to 3.3 x 10(7) pfu/cm(2) for CsCl. The process recovery for the HPLC process was 36% compared to 27.5% for CsCl and total virion to infectious particle ratios of 18 and 11, respectively, were measured. We present a simple two-step chromatography process that is capable of producing high-quality adenovirus at a titre suitable for scale-up and clinical translation.

  8. The evolution of adenoviral vectors through genetic and chemical surface modifications.

    PubMed

    Capasso, Cristian; Garofalo, Mariangela; Hirvinen, Mari; Cerullo, Vincenzo

    2014-02-17

    A long time has passed since the first clinical trial with adenoviral (Ad) vectors. Despite being very promising, Ad vectors soon revealed their limitations in human clinical trials. The pre-existing immunity, the marked liver tropism and the high toxicity of first generation Ad (FG-Ad) vectors have been the main challenges for the development of new approaches. Significant effort toward the development of genetically and chemically modified adenoviral vectors has enabled researchers to create more sophisticated vectors for gene therapy, with an improved safety profile and a higher transduction ability of different tissues. In this review, we will describe the latest findings in the high-speed, evolving field of genetic and chemical modifications of adenoviral vectors, a field in which different disciplines, such as biomaterial research, virology and immunology, co-operate synergistically to create better gene therapy tools for modern challenges.

  9. Adenovirally mediated p53 overexpression diversely influence the cell cycle of HEp-2 and CAL 27 cell lines upon cisplatin and methotrexate treatment.

    PubMed

    Kraljević Pavelić, Sandra; Marjanović, Marko; Poznić, Miroslav; Kralj, Marijeta

    2009-12-01

    p53 gene plays a crucial role in the response to therapy. Since it is inactivated in the majority of human cancers, it is strongly believed that the p53 mutations confer resistance to therapeutics. In this paper we analyzed the influence of two mechanistically diverse antitumor agents--cisplatin and methotrexate on the proliferation and cell cycle of two head and neck squamous cancer cell lines HEp-2 (wild type p53 gene, but HPV 18/E6-inactivated protein) and CAL 27 (mutated p53 gene), along with the influence of adenovirally mediated p53 overexpression in modulation of cisplatin and methoterexate effects, whereby subtoxic vector/compound concentrations were employed. p53 gene was introduced into tumor cells using adenoviral vector (AdCMV-p53). The cell cycle perturbations were measured by two parameter flow cytometry. The expression of p53, p21(WAF1/CIP1) and cyclin B1 proteins was examined using immunocytochemistry and western blot methods. In CAL 27 cells overexpression of p53 completely abrogated high S phase content observed in methotrexate-treated cells into a G1 and slight G2 arrest, while it sustained G2 arrest of the cells treated with cisplatin, along with the reduction of DNA synthesis and cyclin B1 expression. On the other hand, in HEp-2 cell line p53 overexpression slightly slowed down the progression through S phase in cells treated with methotrexate, decreased the cyclin B1 expression only after 24 h, and failed to sustain the G2 arrest after treatment with cisplatin alone. Instead, it increased the population of S phase cells that were not actively synthesizing DNA, sustained cyclin B1 expression and allowed the G2 cells to progress through mitosis. This study demonstrates that adenovirally mediated p53 overexpression at sub-cytotoxic levels enhanced the activity of low doses of cisplatin and methotrexate in HEp-2 and CAL 27 cells through changes in the cell cycle. However, the mechanisms of these effects differ depending on the genetic context and

  10. Regulated Expression of Adenoviral Vectors-Based Gene Therapies

    PubMed Central

    Curtin, James F.; Candolfi, Marianela; Puntel, Mariana; Xiong, Weidong; Muhammad, A. K. M.; Kroeger, Kurt; Mondkar, Sonali; Liu, Chunyan; Bondale, Niyati; Lowenstein, Pedro R.; Castro, Maria G.

    2008-01-01

    Summary Regulatable promoter systems allow gene expression to be tightly controlled in vivo. This is highly desirable for the development of safe, efficacious adenoviral vectors that can be used to treat human diseases in the clinic. Ideally, regulatable cassettes should have minimal gene expression in the “OFF” state, and expression should quickly reach therapeutic levels in the “ON” state. In addition, the components of regulatable cassettes should be non-toxic at physiological concentrations and should not be immunogenic, especially when treating chronic illness that requires long-lasting gene expression. In this chapter, we will describe in detail protocols to develop and validate first generation (Ad) and high-capacity adenoviral (HC-Ad) vectors that express therapeutic genes under the control of the TetON regulatable system. Our laboratory has successfully used these protocols to regulate the expression of marker genes, immune stimulatory genes, and toxins for cancer gene therapeutics, i.e., glioma that is a deadly form of brain cancer. We have shown that this third generation TetON regulatable system, incorporating a doxycycline (DOX)-sensitive rtTA2S-M2 inducer and tTSKid silencer, is non-toxic, relatively non-immunogenic, and can tightly regulate reporter transgene expression downstream of a TRE promoter from adenoviral vectors in vitro and also in vivo. PMID:18470649

  11. Helper-Dependent Adenoviral Vectors.

    PubMed

    Rosewell, Amanda; Vetrini, Francesco; Ng, Philip

    2011-10-29

    Helper-dependent adenoviral vectors are devoid of all viral coding sequences, possess a large cloning capacity, and can efficiently transduce a wide variety of cell types from various species independent of the cell cycle to mediate long-term transgene expression without chronic toxicity. These non-integrating vectors hold tremendous potential for a variety of gene transfer and gene therapy applications. Here, we review the production technologies, applications, obstacles to clinical translation and their potential resolutions, and the future challenges and unanswered questions regarding this promising gene transfer technology.

  12. Helper-Dependent Adenoviral Vectors

    PubMed Central

    Rosewell, Amanda; Vetrini, Francesco; Ng, Philip

    2012-01-01

    Helper-dependent adenoviral vectors are devoid of all viral coding sequences, possess a large cloning capacity, and can efficiently transduce a wide variety of cell types from various species independent of the cell cycle to mediate long-term transgene expression without chronic toxicity. These non-integrating vectors hold tremendous potential for a variety of gene transfer and gene therapy applications. Here, we review the production technologies, applications, obstacles to clinical translation and their potential resolutions, and the future challenges and unanswered questions regarding this promising gene transfer technology. PMID:24533227

  13. Genetically engineering adenoviral vectors for gene therapy.

    PubMed

    Coughlan, Lynda

    2014-01-01

    Adenoviral (Ad) vectors are commonly used for various gene therapy applications. Significant advances in the genetic engineering of Ad vectors in recent years has highlighted their potential for the treatment of metastatic disease. There are several methods to genetically modify the Ad genome to incorporate retargeting peptides which will redirect the natural tropism of the viruses, including homologous recombination in bacteria or yeast. However, homologous recombination in yeast is highly efficient and can be achieved without the need for extensive cloning strategies. In addition, the method does not rely on the presence of unique restriction sites within the Ad genome and the reagents required for this method are widely available and inexpensive. Large plasmids containing the entire adenoviral genome (~36 kbp) can be modified within Saccharomyces cerevisiae yeast and genomes easily rescued in Escherichia coli hosts for analysis or amplification. A method for two-step homologous recombination in yeast is described in this chapter.

  14. Evaluation of signal transduction pathways after transient cutaneous adenoviral gene delivery

    PubMed Central

    2011-01-01

    Background Adenoviral vectors have provided effective methods for in vivo gene delivery in therapeutic applications. However, these vectors can induce immune responses that may severely affect the ability of vector re-application. There is limited information about the mechanisms and signal transduction pathways involved in adenoviral recognition. For optimization of cutaneous gene therapy it is necessary to investigate molecular mechanisms of virus recognition in epidermal cells. The aim of this study was to investigate the signal transduction of the innate immunity after adenoviral DNA internalization in keratinocytes. Methods In vitro, keratinocytes were transfected with DNA, in the presence and absence of inhibitors for signalling molecules. In vivo, immunocompetent and athymic mice (n = 3 per group) were twice transduced with an Ad-vector. Results The results show an acute induction of type-I-interferon after in vitro transfection. Inhibition of PI3K, p38 MAPK, JNK and NFkappaB resulted in a decreased expression of type-I-interferon. In contrast to immunocompetent mice, athymic mice demonstrated a constant transgene expression and reduced inflammatory response in vivo. Conclusion The results suggest an induction of the innate immunity triggered by cytoplasm localised DNA which is mediated by PI3K-, p38 MAPK-, JNK-, NFkappaB-, JAK/STAT- and ERK1/2-dependent pathways. A stable transgene expression and a reduced inflammatory response in immunodeficient mice have been observed. These results provide potential for an effective adenoviral gene delivery into immunosupressed skin. PMID:21255430

  15. Enhanced radiosensitivity of malignant glioma cells after adenoviral p53 transduction.

    PubMed

    Broaddus, W C; Liu, Y; Steele, L L; Gillies, G T; Lin, P S; Loudon, W G; Valerie, K; Schmidt-Ullrich, R K; Fillmore, H L

    1999-12-01

    The goal of this study was to determine whether adenoviral vector-mediated expression of human wildtype p53 can enhance the radiosensitivity of malignant glioma cells that express native wild-type p53. The p53 gene is thought to function abnormally in the majority of malignant gliomas, although it has been demonstrated to be mutated in only approximately 30%. This has led to studies in which adenoviral transduction with wild-type human p53 has been investigated in an attempt to slow tumor cell growth. Recent studies suggest that reconstitution of wild-type p53 can render cells more susceptible to radiation-mediated death, primarily by p53-mediated apoptosis. Rat RT2 glioma cells were analyzed for native p53 status by reverse transcriptase-polymerase chain reaction and sequence analysis and for p53 expression by Western blot analysis. Clonogenic survival and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assay were used to characterize RT2 cell radiosensitivity and apoptosis, respectively, with and without prior transduction with p53-containing and control adenoviral vectors. Animal survival length was monitored after intracerebral implantation with transduced and nontransduced RT2 cells, with and without cranial radiation. The RT2 cells were demonstrated to express native rat wild-type p53 and to markedly overexpress human p53 following adenoviral p53 transduction. The combination of p53 transduction followed by radiation resulted in marked decreases in RT2 cell survival and increases in apoptosis at radiation doses from 2 to 6 Gy. Animals receiving cranial radiation after intracerebral implantation with RT2 cells previously transduced with p53 survived significantly longer than control animals (p<0.01). The ability to enhance the radiosensitivity of malignant glioma cells that express wild-type p53 by using adenoviral transduction to induce overexpression of p53 offers hope for this approach as a therapeutic strategy

  16. Helper-dependent adenoviral vectors for liver-directed gene therapy

    PubMed Central

    Brunetti-Pierri, Nicola; Ng, Philip

    2011-01-01

    Helper-dependent adenoviral (HDAd) vectors devoid of all viral-coding sequences are promising non-integrating vectors for liver-directed gene therapy because they have a large cloning capacity, can efficiently transduce a wide variety of cell types from various species independent of the cell cycle and can result in long-term transgene expression without chronic toxicity. The main obstacle preventing clinical applications of HDAd for liver-directed gene therapy is the host innate inflammatory response against the vector capsid proteins that occurs shortly after intravascular vector administration resulting in acute toxicity, the severity of which is dependent on vector dose. Intense efforts have been focused on elucidating the factors involved in this acute response and various strategies have been investigated to improve the therapeutic index of HDAd vectors. These strategies have yielded encouraging results with the potential for clinical translation. PMID:21470977

  17. Microsphere-liposome complexes protect adenoviral vectors from neutralising antibody without losses in transfection efficiency, in-vitro.

    PubMed

    Steel, Jason C; Cavanagh, Heather M A; Burton, Mark A; Kalle, Wouter H J

    2004-11-01

    Adenoviral vectors have been commonly used in gene therapy protocols but the success of their use is often limited by the induction of host immunity to the vector. Following exposure to the adenoviral vector, adenoviral-specific neutralising antibodies are produced, which limits further administration. This study examines the effectiveness of a novel combination of microspheres and liposomes for the shielding of adenovirus from neutralising antibodies in an in-vitro setting. We show that liposomes are effective in the protection of adenovirus from neutralising antibody and that the conjugation of these complexes to microspheres augments the level of protection. This study further reveals that previously neutralised adenovirus may still be transported into the cell via liposome-cell interactions and is still capable of expressing its genes, making this vector an effective tool for circumvention of the humoral immune response. We also looked at possible side effects of using the complexes, namely increases in cytotoxicity and reductions in transfection efficiency. Our results showed that varying the liposome:adenovirus ratio can reduce the cytotoxicity of the vector as well as increase the transfection efficiency. In addition, in cell lines that are adenoviral competent, transfection efficiencies on par with uncomplexed adenoviral vectors were achievable with the combination vector.

  18. Treatment of Adenoviral Acute Respiratory Distress Syndrome Using Cidofovir With Extracorporeal Membrane Oxygenation.

    PubMed

    Lee, Minhyeok; Kim, Seulgi; Kwon, Oh Jung; Kim, Ji Hye; Jeong, Inbeom; Son, Ji Woong; Na, Moon Jun; Yoon, Yoo Sang; Park, Hyun Woong; Kwon, Sun Jung

    2017-03-01

    Adenovirus infections are associated with respiratory (especially upper respiratory) infection and gastrointestinal disease and occur primarily in infants and children. Although rare in adults, severe lower respiratory adenovirus infections including pneumonia are reported in specific populations, such as military recruits and immunocompromised patients. Antiviral treatment is challenging due to limited clinical experience and lack of well-controlled randomized trials. Several previously reported cases of adenoviral pneumonia showed promising efficacy of cidofovir. However, few reports discussed the efficacy of cidofovir in acute respiratory distress syndrome (ARDS). We experienced 3 cases of adenoviral pneumonia associated with ARDS and treated with cidofovir and respiratory support, including extracorporeal membrane oxygenation (ECMO). All 3 patients showed a positive clinical response to cidofovir and survival at 28 days. Cidofovir with early ECMO therapy may be a therapeutic option in adenoviral ARDS. A literature review identified 15 cases of adenovirus pneumonia associated with ARDS.

  19. Discovering Conformational Sub-States Relevant to Protein Function

    PubMed Central

    Ramanathan, Arvind; Savol, Andrej J.; Langmead, Christopher J.; Agarwal, Pratul K.; Chennubhotla, Chakra S.

    2011-01-01

    Background Internal motions enable proteins to explore a range of conformations, even in the vicinity of native state. The role of conformational fluctuations in the designated function of a protein is widely debated. Emerging evidence suggests that sub-groups within the range of conformations (or sub-states) contain properties that may be functionally relevant. However, low populations in these sub-states and the transient nature of conformational transitions between these sub-states present significant challenges for their identification and characterization. Methods and Findings To overcome these challenges we have developed a new computational technique, quasi-anharmonic analysis (QAA). QAA utilizes higher-order statistics of protein motions to identify sub-states in the conformational landscape. Further, the focus on anharmonicity allows identification of conformational fluctuations that enable transitions between sub-states. QAA applied to equilibrium simulations of human ubiquitin and T4 lysozyme reveals functionally relevant sub-states and protein motions involved in molecular recognition. In combination with a reaction pathway sampling method, QAA characterizes conformational sub-states associated with cis/trans peptidyl-prolyl isomerization catalyzed by the enzyme cyclophilin A. In these three proteins, QAA allows identification of conformational sub-states, with critical structural and dynamical features relevant to protein function. Conclusions Overall, QAA provides a novel framework to intuitively understand the biophysical basis of conformational diversity and its relevance to protein function. PMID:21297978

  20. Administration of helper-dependent adenoviral vectors and sequential delivery of different vector serotype for long-term liver-directed gene transfer in baboons

    PubMed Central

    Morral, Núria; O’Neal, Wanda; Rice, Karen; Leland, Michele; Kaplan, Johanne; Piedra, Pedro A.; Zhou, Heshan; Parks, Robin J.; Velji, Rizwan; Aguilar-Córdova, Estuardo; Wadsworth, Samuel; Graham, Frank L.; Kochanek, Stefan; Carey, K. Dee; Beaudet, Arthur L.

    1999-01-01

    The efficiency of first-generation adenoviral vectors as gene delivery tools is often limited by the short duration of transgene expression, which can be related to immune responses and to toxic effects of viral proteins. In addition, readministration is usually ineffective unless the animals are immunocompromised or a different adenovirus serotype is used. Recently, adenoviral vectors devoid of all viral coding sequences (helper-dependent or gutless vectors) have been developed to avoid expression of viral proteins. In mice, liver-directed gene transfer with AdSTK109, a helper-dependent adenoviral (Ad) vector containing the human α1-antitrypsin (hAAT) gene, resulted in sustained expression for longer than 10 months with negligible toxicity to the liver. In the present report, we have examined the duration of expression of AdSTK109 in the liver of baboons and compared it to first-generation vectors expressing hAAT. Transgene expression was limited to approximately 3–5 months with the first-generation vectors. In contrast, administration of AdSTK109 resulted in transgene expression for longer than a year in two of three baboons. We have also investigated the feasibility of circumventing the humoral response to the virus by sequential administration of vectors of different serotypes. We found that the ineffectiveness of readministration due to the humoral response to an Ad5 first-generation vector was overcome by use of an Ad2-based vector expressing hAAT. These data suggest that long-term expression of transgenes should be possible by combining the reduced immunogenicity and toxicity of helper-dependent vectors with sequential delivery of vectors of different serotypes. PMID:10536005

  1. Adenoviral receptor expression of normal bladder and transitional cell carcinoma of the bladder.

    PubMed

    Buscarini, Maurizio; Quek, Marcus L; Gilliam-Hegarich, Susan; Kasahara, Nori; Bochner, Bernard

    2007-01-01

    The insertion of absent or underexpressed genes into cancer cells to alter their malignant phenotype is an important potential application of available gene therapy technology. One of the more common viral vector systems that has been extensively studied for this purpose are the replication-deficient adenoviruses (Ad). Adenoviral infection of cells is mediated through a complex pathway, initiated following viral-cell attachment. Adenoviral-cell attachment occurs following interactions with a 46-kDa transmembrane protein with high affinity for both the Coxsackie and adenovirus, designated the CAR (Coxsackie and adenoviral receptor). Additional important cell-viral interactions that occur involve the alpha(v)-based integrins, specifically alpha(v)beta3 and alpha(v)beta5. The purpose of the present study was to determine the extent of expression and localization of the known Ad receptor proteins (CAR, alpha(v)beta3, and alpha(v)beta5) in normal and cancerous human bladders. Frozen tissue samples of normal bladder and invasive transitional cell cancers of the bladder were evaluated. Tissue blocks containing muscle-invasive transitional cell carcinoma (TCC) were obtained following radical cystectomy, which were performed at our institution. Thirty-two invasive transitional cell bladder tumors were evaluated, each with a matched sample of histologically normal-appearing bladder used as a control. Four additional samples of normal bladder were obtained from patients with no evidence of disease of the bladder and served as further controls. Three additional cases of invasive bladder cancer with no matching normal tissue were also evaluated. Identification of the CAR receptor was performed using the anti-CAR mouse monoclonal antibody designated RmBC. The integrins alpha(v)beta3 and alpha(v)beta5 were identified using the mouse monoclonal antibodies designated LM609 and P1F6 respectively. All slides were evaluated by two of the authors (M.B., B.B.) without knowledge of the

  2. Adenoviral infection in a collection of juvenile inland bearded dragons (Pogona vitticeps).

    PubMed

    Doneley, R J T; Buckle, K N; Hulse, L

    2014-01-01

    Juvenile inland bearded dragons (Pogona vitticeps) from a breeding collection in south-east Queensland were presented at age 6-10 weeks with neurological signs, poor growth and occasional deaths. Histopathological examination revealed that six of eight lizards had multifocal non-suppurative hepatitis associated with 5-10 μm diameter, smudgy, basophilic, hyaline intranuclear inclusion bodies that marginated the nuclear chromatin. These histological lesions were considered consistent with adenoviral hepatitis. Infection with adenovirus was confirmed positive in one of the eight dragons by PCR for adenoviral DNA. DNA was extracted from formalin-fixed paraffin-embedded pooled tissues of the juvenile inland bearded dragons and tested using a nested-PCR protocol with primers specific for identification of adenovirus. Sequencing of the one PCR-positive dragon showed 95% nucleotide sequence alignment with agamid atadenovirus 1. Further investigation involved testing the breeding population, including the parents of the affected juveniles. Blood and cloacal samples were collected from the adult population, DNA was extracted and tested by PCR for adenovirus. There was a high percentage of positive results from the samples collected from the breeding population. This is the first reported group outbreak of adenoviral disease in bearded dragons in Australia. © 2014 Australian Veterinary Association.

  3. Functional characterization of adenoviral/retroviral chimeric vectors and their use for efficient screening of retroviral producer cell lines.

    PubMed

    Duisit, G; Salvetti, A; Moullier, P; Cosset, F L

    1999-01-20

    We have generated three different E1-deleted replication-defective adenoviral vectors expressing either Moloney murine leukemia virus (Mo-MuLV) Gag-Pol core particle proteins, gibbon ape leukemia virus (GALV) envelope glycoproteins, or an MuLV-derived retroviral vector genome encoding mCD2 antigen, a murine cell surface marker easily detectable by flow cytometry. Each of the three vectors was first characterized individually by infection of cells providing the complementary retroviral function(s) and able to induce the production of retroviral vectors with an efficiency similar to or higher than that of FLY stable retroviral packaging cells [Cosset, F.-L., Takeuchi, Y., Battini, J.-L., Weiss, R.A., and Collins, M.K.L., (1995). J. Virol. 69, 7430-7436]. In small-scale pilot experiments, TE671 cells simultaneously coinfected with the three adenoviral vectors efficiently released helper-free retroviral vectors in their supernatant, with titers greater than 10(6) infectious particles per milliliter by end-point titrations. Our results also indicated that in contrast to retroviral vector-packageable RNAs, the adenovirus-mediated overexpression of both Gag-Pol and Env packaging functions had limited impact on retroviral titers. The primary mechanism suspected is the premature intracellular cleavage of the Pr65gag precursor that we found in gag-pol-expressing cells, which in turn may impair the normal incorporation of high loads of functional Env. Last, the characterization of the adenoviral/retroviral chimeric vectors allowed the screening of various primate cells for retroviral production and we found that three hepatocyte-derived cell lines were highly efficient in the assembly and release of infectious retroviral particles.

  4. Increased tumor localization and reduced immune response to adenoviral vector formulated with the liposome DDAB/DOPE.

    PubMed

    Steel, Jason C; Cavanagh, Heather M A; Burton, Mark A; Abu-Asab, Mones S; Tsokos, Maria; Morris, John C; Kalle, Wouter H J

    2007-04-01

    We aimed to increase the efficiency of adenoviral vectors by limiting adenoviral spread from the target site and reducing unwanted host immune responses to the vector. We complexed adenoviral vectors with DDAB-DOPE liposomes to form adenovirus-liposomal (AL) complexes. AL complexes were delivered by intratumoral injection in an immunocompetent subcutaneous rat tumor model and the immunogenicity of the AL complexes and the expression efficiency in the tumor and other organs was examined. Animals treated with the AL complexes had significantly lower levels of beta-galactosidase expression in systemic tissues compared to animals treated with the naked adenovirus (NA) (P<0.05). The tumor to non-tumor ratio of beta-galactosidase marker expression was significantly higher for the AL complex treated animals. NA induced significantly higher titers of adenoviral-specific antibodies compared to the AL complexes (P<0.05). The AL complexes provided protection (immunoshielding) to the adenovirus from neutralizing antibody. Forty-seven percent more beta-galactosidase expression was detected following intratumoral injection with AL complexes compared to the NA in animals pre-immunized with adenovirus. Complexing of adenovirus with liposomes provides a simple method to enhance tumor localization of the vector, decrease the immunogenicity of adenovirus, and provide protection of the virus from pre-existing neutralizing antibodies.

  5. Adenovirus Type 5 Viral Particles Pseudotyped with Mutagenized Fiber Proteins Show Diminished Infectivity of Coxsackie B-Adenovirus Receptor-Bearing Cells

    PubMed Central

    Jakubczak, John L.; Rollence, Michele L.; Stewart, David A.; Jafari, Jonathon D.; Von Seggern, Dan J.; Nemerow, Glen R.; Stevenson, Susan C.; Hallenbeck, Paul L.

    2001-01-01

    A major limitation of adenovirus type 5 (Ad5)-based gene therapy, the inability to target therapeutic genes to selected cell types, is attributable to the natural tropism of the virus for the widely expressed coxsackievirus-adenovirus receptor (CAR) protein. Modifications of the Ad5 fiber knob domain have been shown to alter the tropism of the virus. We have developed a novel system to rapidly evaluate the function of modified fiber proteins in their most relevant context, the adenoviral capsid. This transient transfection/infection system combines transfection of cells with plasmids that express high levels of the modified fiber protein and infection with Ad5.βgal.ΔF, an E1-, E3-, and fiber-deleted adenoviral vector encoding β-galactosidase. We have used this system to test the adenoviral transduction efficiency mediated by a panel of fiber protein mutants that were proposed to influence CAR interaction. A series of amino acid modifications were incorporated via mutagenesis into the fiber expression plasmid, and the resulting fiber proteins were subsequently incorporated onto adenoviral particles. Mutations located in the fiber knob AB and CD loops demonstrated the greatest reduction in fiber-mediated gene transfer in HeLa cells. We also observed effects on transduction efficiency with mutations in the FG loop, indicating that the binding site may extend to the adjacent monomer in the fiber trimer and in the HI loop. These studies support the concept that modification of the fiber knob domain to diminish or ablate CAR interaction should result in a detargeted adenoviral vector that can be combined simultaneously with novel ligands for the development of a systemically administered, targeted adenoviral vector. PMID:11222722

  6. Recombinant HT{sub m4} gene, protein and assays

    DOEpatents

    Lim, B.; Adra, C.N.; Lelias, J.M.

    1996-09-03

    The invention relates to a recombinant DNA molecule which encodes a HT{sub m4} protein, a transformed host cell which has been stably transfected with a DNA molecule which encodes a HT{sub m4} protein and a recombinant HT{sub m4} protein. The invention also relates to a method for detecting the presence of a hereditary atopy. 2 figs.

  7. Recombinant HT.sub.m4 gene, protein and assays

    DOEpatents

    Lim, Bing; Adra, Chaker N.; Lelias, Jean-Michel

    1996-01-01

    The invention relates to a recombinant DNA molecule which encodes a HT.sub.m4 protein, a transformed host cell which has been stably transfected with a DNA molecule which encodes a HT.sub.m4 protein and a recombinant HT.sub.m4 protein. The invention also relates to a method for detecting the presence of a hereditary atopy.

  8. Adenovirally transduced bone marrow stromal cells differentiate into pigment epithelial cells and induce rescue effects in RCS rats.

    PubMed

    Arnhold, Stefan; Heiduschka, Peter; Klein, Helmut; Absenger, Yvonne; Basnaoglu, Serkan; Kreppel, Florian; Henke-Fahle, Sylvia; Kochanek, Stefan; Bartz-Schmidt, Karl-Ulrich; Addicks, Klaus; Schraermeyer, Ulrich

    2006-09-01

    To determine the potential of adenovirally transduced bone marrow stromal cells (BMSCs) to differentiate into retinal pigment epithelial-like cells and to evaluabe possible rescue effects after transplantation into the retinas of Royal College of Surgeons (RCS) rats. Through a high-capacity adenoviral vector expressing either green fluorescent protein (GFP) or pigment epithelial-derived factor (PEDF), rat MSCs were transduced in vitro before subretinal transplantation into Wistar rats or, alternatively, RCS rats. Two months after cell injection, the rats were killed and the eyes enucleated. The eyes were then investigated light microscopically or processed for electron microscopic investigations. Cell differentiation and integration were analyzed immunocytochemically using antibodies against cytokeratin and the tight junction protein ZO-1. Electroretinography was performed 16 days after injection of cells, to check whether a functional rescue could be detected. In vitro experiments in cocultured human MSCs and human RPE cells showed that MSCs adopted RPE-like characteristics. In grafting experiments, some rat MSCs integrate into the host RPE cell layer of Wistar and RCS rats, indicated by their hexagonal morphology. Subretinally transplanted cells express the epithelial marker cytokeratin and establish tight junctions with the host RPE cells. Furthermore, rescue effects can be demonstrated after grafting of vector-transduced and nontransduced MSCs in semithin sections of dystrophic retinas. Ultrastructurally, MSCs can be detected on top of host RPE and in close contact with photoreceptor outer segments phagocytosing rod outer segments. Taken together, these results raise the possibility that MSCs have the potency to replace diseased RPE cells and deliver therapeutic proteins into the subretinal space to protect photoreceptor cells from degeneration.

  9. Protein Sub-Nuclear Localization Prediction Using SVM and Pfam Domain Information

    PubMed Central

    Kumar, Ravindra; Jain, Sohni; Kumari, Bandana; Kumar, Manish

    2014-01-01

    The nucleus is the largest and the highly organized organelle of eukaryotic cells. Within nucleus exist a number of pseudo-compartments, which are not separated by any membrane, yet each of them contains only a specific set of proteins. Understanding protein sub-nuclear localization can hence be an important step towards understanding biological functions of the nucleus. Here we have described a method, SubNucPred developed by us for predicting the sub-nuclear localization of proteins. This method predicts protein localization for 10 different sub-nuclear locations sequentially by combining presence or absence of unique Pfam domain and amino acid composition based SVM model. The prediction accuracy during leave-one-out cross-validation for centromeric proteins was 85.05%, for chromosomal proteins 76.85%, for nuclear speckle proteins 81.27%, for nucleolar proteins 81.79%, for nuclear envelope proteins 79.37%, for nuclear matrix proteins 77.78%, for nucleoplasm proteins 76.98%, for nuclear pore complex proteins 88.89%, for PML body proteins 75.40% and for telomeric proteins it was 83.33%. Comparison with other reported methods showed that SubNucPred performs better than existing methods. A web-server for predicting protein sub-nuclear localization named SubNucPred has been established at http://14.139.227.92/mkumar/subnucpred/. Standalone version of SubNucPred can also be downloaded from the web-server. PMID:24897370

  10. Randomized, Controlled, Phase 2 Trial of Povidone-Iodine/Dexamethasone Ophthalmic Suspension for Treatment of Adenoviral Conjunctivitis.

    PubMed

    Pepose, Jay S; Ahuja, Arjun; Liu, Wenlei; Narvekar, Abhijit; Haque, Reza

    2018-05-19

    To evaluate the efficacy/safety of an ophthalmic suspension of povidone-iodine (PVP-I) 0.6% and dexamethasone 0.1% in patients with acute adenoviral conjunctivitis. Multicenter, randomized, vehicle-controlled, double-masked trial. Adults with a positive Rapid Pathogen Screening Adeno-Detector Plus TM test were randomized 1:1:1 to PVP-I 0.6%/dexamethasone 0.1%, PVP-I 0.6%, or vehicle, bilaterally 4 times daily for 5 days (days 1-5). Patients were evaluated on days 3, 6, and 12 (+1-day window). Efficacy measures included clinical resolution and adenoviral eradication. Overall, 144 patients were included in the efficacy analysis (PVP-I/dexamethasone, n = 48; PVP-I, n = 50; vehicle, n = 46). The proportion of patients with clinical resolution (primary study eye with last observation carried forward [LOCF]) at the day 6 visit was higher with PVP-I/dexamethasone (31.3%) than vehicle (10.9%; P = .0158) and PVP-I (18.0%; P = nonsignificant). The proportion with adenoviral eradication (primary study eye with LOCF) was higher with PVP-I/dexamethasone than vehicle at the day 3 (35.4% vs 8.7%; P = .0019) and day 6 (79.2% vs 56.5%; P = .0186) visits and versus PVP-I (day 3 visit, 32.0%; day 6 visit, 62.0%; each P = nonsignificant). Treatment-emergent adverse events (AEs) occurred in 69.0% (vehicle), 62.7% (PVP-I), and 53.4% (PVP-I/dexamethasone) of patients in the safety dataset. Discontinuation owing to AEs occurred in 37 patients (vehicle, n = 16; PVP-I, n = 12; PVP-I/dexamethasone, n = 9). PVP-I/dexamethasone appeared safe and well tolerated, and significantly improved clinical resolution and adenoviral eradication in patients with acute adenoviral conjunctivitis. Copyright © 2018. Published by Elsevier Inc.

  11. The dimeric [V{sub 2}O{sub 2}F{sub 8}]{sup 4−} anion: Structural characterization of a magnetic basic-building-unit

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

    Lu, Hongcheng, E-mail: hchlu@northwestern.edu; State Key Laboratory of Solidification Processing, School of Material Science and Engineering, Northwestern Polytechnical University, 127 Youyixilu Road, Xi'an 710072; Gautier, Romain, E-mail: r-gautier@northwestern.edu

    2013-04-15

    New materials built from the [V{sub 2}O{sub 2}F{sub 8}]{sup 4−} anionic basic-building-unit (BBU) exhibit interesting magnetic properties owing to the proximity of the two d{sup 1} V(IV) cations and the orbital interactions of fluoride and oxide ligands. In our search to target such materials, the vanadium oxide–fluoride compound [dpaH{sub 2}]{sub 2}[V{sub 2}O{sub 2}F{sub 8}] in which a dimeric anion [V{sub 2}O{sub 2}F{sub 8}]{sup 4−} is isolated in a hydrogen bond network was hydrothermally synthesized (dpa=2,2′-dipyridylamine). This hydrogen bond network is able to stabilize the highly ionic species [V{sub 2}O{sub 2}F{sub 8}]{sup 4−} as demonstrated with bond valence calculations. The coordinationmore » of the O{sup 2−}/F{sup −} ordered ligands was investigated and antiferromagnetic coupling of the isolated BBU was measured. - The new hybrid compound [dpaH{sub 2}]{sub 2}[V{sub 2}O{sub 2}F{sub 8}] built from the interesting [V{sub 2}O{sub 2}F{sub 8}]{sup 4−} magnetic basic-building-unit (BBU) was synthesized by the hydrothermal method. The coordination of the O{sup 2−}/F{sup −} ordered ligands was investigated by BVS calculations and antiferromagnetic coupling was measured. Highlights: ► A new vanadium oxyfluoride was synthesized by hydrothermal method. ► The Dimeric [V{sub 2}O{sub 2}F{sub 8}]{sup 4−} basic building unit is isolated in the hydrogen bond networks. ► The coordination of [V{sub 2}O{sub 2}F{sub 8}]{sup 4−} units to the extended structure is investigated. ► Isolated [V{sub 2}O{sub 2}F{sub 8}]{sup 4−} units exhibit antiferromagnetic coupling.« less

  12. A clinical algorithm identifies high risk pediatric oncology and bone marrow transplant patients likely to benefit from treatment of adenoviral infection.

    PubMed

    Williams, Kirsten Marie; Agwu, Allison L; Dabb, Alix A; Higman, Meghan A; Loeb, David M; Valsamakis, Alexandra; Chen, Allen R

    2009-11-01

    Adenoviral infections cause morbidity and mortality in blood and marrow transplantation and pediatric oncology patients. Cidofovir is active against adenovirus, but must be used judiciously because of its nephrotoxicity and unclear indications. Therefore, before introducing cidofovir use during an adenoviral outbreak, we developed a clinical algorithm to distinguish low risk patients from those who merited cidofovir therapy because of significant adenoviral disease and high risk for death. This study was conducted to determine whether the algorithm accurately predicted severe adenovirus disease and whether selective cidofovir treatment was beneficial. A retrospective analysis of a pediatric oncology/blood and marrow transplantation cohort prealgorithm and postalgorithm implementation was performed. Twenty patients with adenovirus infection were identified (14 high risk and 6 low risk). All low-risk patients cleared their infections without treatment. Before algorithm implementation, all untreated high-risk patients died, 4 out of 5 (80%), from adenoviral infection. In contrast, cidofovir reduced adenovirus-related mortality in the high-risk group postalgorithm implementation (9 patients treated, 1 patient died; RR 0.14, P<0.05) and all treated high-risk patients cleared their virus. The clinical algorithm accurately identified patients at high risk for severe fatal adenoviral disease who would benefit from selective use of cidofovir.

  13. High Efficiency CRISPR/Cas9-mediated Gene Editing in Primary Human T-cells Using Mutant Adenoviral E4orf6/E1b55k "Helper" Proteins.

    PubMed

    Gwiazda, Kamila S; Grier, Alexandra E; Sahni, Jaya; Burleigh, Stephen M; Martin, Unja; Yang, Julia G; Popp, Nicholas A; Krutein, Michelle C; Khan, Iram F; Jacoby, Kyle; Jensen, Michael C; Rawlings, David J; Scharenberg, Andrew M

    2016-09-29

    Many future therapeutic applications of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 and related RNA-guided nucleases are likely to require their use to promote gene targeting, thus necessitating development of methods that provide for delivery of three components-Cas9, guide RNAs and recombination templates-to primary cells rendered proficient for homology-directed repair. Here, we demonstrate an electroporation/transduction codelivery method that utilizes mRNA to express both Cas9 and mutant adenoviral E4orf6 and E1b55k helper proteins in association with adeno-associated virus (AAV) vectors expressing guide RNAs and recombination templates. By transiently enhancing target cell permissiveness to AAV transduction and gene editing efficiency, this novel approach promotes efficient gene disruption and/or gene targeting at multiple loci in primary human T-cells, illustrating its broad potential for application in translational gene editing.

  14. SChloro: directing Viridiplantae proteins to six chloroplastic sub-compartments.

    PubMed

    Savojardo, Castrense; Martelli, Pier Luigi; Fariselli, Piero; Casadio, Rita

    2017-02-01

    Chloroplasts are organelles found in plants and involved in several important cell processes. Similarly to other compartments in the cell, chloroplasts have an internal structure comprising several sub-compartments, where different proteins are targeted to perform their functions. Given the relation between protein function and localization, the availability of effective computational tools to predict protein sub-organelle localizations is crucial for large-scale functional studies. In this paper we present SChloro, a novel machine-learning approach to predict protein sub-chloroplastic localization, based on targeting signal detection and membrane protein information. The proposed approach performs multi-label predictions discriminating six chloroplastic sub-compartments that include inner membrane, outer membrane, stroma, thylakoid lumen, plastoglobule and thylakoid membrane. In comparative benchmarks, the proposed method outperforms current state-of-the-art methods in both single- and multi-compartment predictions, with an overall multi-label accuracy of 74%. The results demonstrate the relevance of the approach that is eligible as a good candidate for integration into more general large-scale annotation pipelines of protein subcellular localization. The method is available as web server at http://schloro.biocomp.unibo.it gigi@biocomp.unibo.it.

  15. HIV-1 adenoviral vector vaccines expressing multi-trimeric BAFF and 4-1BBL enhance T cell mediated anti-viral immunity.

    PubMed

    Kanagavelu, Saravana; Termini, James M; Gupta, Sachin; Raffa, Francesca N; Fuller, Katherine A; Rivas, Yaelis; Philip, Sakhi; Kornbluth, Richard S; Stone, Geoffrey W

    2014-01-01

    Adenoviral vectored vaccines have shown considerable promise but could be improved by molecular adjuvants. Ligands in the TNF superfamily (TNFSF) are potential adjuvants for adenoviral vector (Ad5) vaccines based on their central role in adaptive immunity. Many TNFSF ligands require aggregation beyond the trimeric state (multi-trimerization) for optimal biological function. Here we describe Ad5 vaccines for HIV-1 Gag antigen (Ad5-Gag) adjuvanted with the TNFSF ligands 4-1BBL, BAFF, GITRL and CD27L constructed as soluble multi-trimeric proteins via fusion to Surfactant Protein D (SP-D) as a multimerization scaffold. Mice were vaccinated with Ad5-Gag combined with Ad5 expressing one of the SP-D-TNFSF constructs or single-chain IL-12p70 as adjuvant. To evaluate vaccine-induced protection, mice were challenged with vaccinia virus expressing Gag (vaccinia-Gag) which is known to target the female genital tract, a major route of sexually acquired HIV-1 infection. In this system, SP-D-4-1BBL or SP-D-BAFF led to significantly reduced vaccinia-Gag replication when compared to Ad5-Gag alone. In contrast, IL-12p70, SP-D-CD27L and SP-D-GITRL were not protective. Histological examination following vaccinia-Gag challenge showed a dramatic lymphocytic infiltration into the uterus and ovaries of SP-D-4-1BBL and SP-D-BAFF-treated animals. By day 5 post challenge, proinflammatory cytokines in the tissue were reduced, consistent with the enhanced control over viral replication. Splenocytes had no specific immune markers that correlated with protection induced by SP-D-4-1BBL and SP-D-BAFF versus other groups. IL-12p70, despite lack of anti-viral efficacy, increased the total numbers of splenic dextramer positive CD8+ T cells, effector memory T cells, and effector Gag-specific CD8+ T cells, suggesting that these markers are poor predictors of anti-viral immunity in this model. In conclusion, soluble multi-trimeric 4-1BBL and BAFF adjuvants led to strong protection from vaccinia

  16. CRISPR/Cas9 delivery with one single adenoviral vector devoid of all viral genes.

    PubMed

    Ehrke-Schulz, Eric; Schiwon, Maren; Leitner, Theo; Dávid, Stephan; Bergmann, Thorsten; Liu, Jing; Ehrhardt, Anja

    2017-12-07

    The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system revolutionized the field of gene editing but viral delivery of the CRISPR/Cas9 system has not been fully explored. Here we adapted clinically relevant high-capacity adenoviral vectors (HCAdV) devoid of all viral genes for the delivery of the CRISPR/Cas9 machinery using a single viral vector. We present a platform enabling fast transfer of the Cas9 gene and gRNA expression units into the HCAdV genome including the option to choose between constitutive or inducible Cas9 expression and gRNA multiplexing. Efficacy and versatility of this pipeline was exemplified by producing different CRISPR/Cas9-HCAdV targeting the human papillomavirus (HPV) 18 oncogene E6, the dystrophin gene causing Duchenne muscular dystrophy (DMD) and the HIV co-receptor C-C chemokine receptor type 5 (CCR5). All CRISPR/Cas9-HCAdV proved to be efficient to deliver the respective CRISPR/Cas9 expression units and to introduce the desired DNA double strand breaks at their intended target sites in immortalized and primary cells.

  17. Determinants of Outcomes of Adenoviral Keratoconjunctivitis.

    PubMed

    Lee, Cecilia S; Lee, Aaron Y; Akileswaran, Lakshmi; Stroman, David; Najafi-Tagol, Kathryn; Kleiboeker, Steve; Chodosh, James; Magaret, Amalia; Wald, Anna; Van Gelder, Russell N

    2018-03-27

    To determine host and pathogen factors predictive of outcomes in a large clinical cohort with keratoconjunctivitis. Retrospective analyses of the clinical and molecular data from a randomized, controlled, masked trial for auricloscene for keratoconjunctivitis (NVC-422 phase IIB, NovaBay; clinicaltrials.gov identifier, NCT01877694). Five hundred participants from United States, India, Brazil, and Sri Lanka with clinical diagnosis of keratoconjunctivitis and positive rapid test results for adenovirus. Clinical signs and symptoms and bilateral conjunctival swabs were obtained on days 1, 3, 6, 11, and 18. Polymerase chain reaction (PCR) analysis was performed to detect and quantify adenovirus in all samples. Regression models were used to evaluate the association of various variables with keratoconjunctivitis outcomes. Time to resolution of each symptom or sign was assessed by adenoviral species with Cox regression. The difference in composite scores of clinical signs between days 1 and 18, mean visual acuity change between days 1 and 18, and time to resolution of each symptom or sign. Of 500 participants, 390 (78%) showed evidence of adenovirus by PCR. Among adenovirus-positive participants, adenovirus D species was most common (63% of total cases), but a total of 4 species and 21 different types of adenovirus were detected. Adenovirus D was associated with more severe signs and symptoms, a higher rate of subepithelial infiltrate development, and a slower decline in viral load compared with all other adenovirus species. The clinical courses of all patients with non-adenovirus D species infection and adenovirus-negative keratoconjunctivitis were similar. Mean change in visual acuity between days 1 and 18 was a gain of 1.9 letters; worse visual outcome was associated with older age. A substantial proportion of keratoconjunctivitis is not associated with a detectable adenovirus. The clinical course of those with adenovirus D keratoconjunctivitis is significantly more

  18. 7 CFR 275.7 - Selection of sub-units for review.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... situations, the sub-unit would be classified based upon its primary function exclusively. However, when any... classification and if selected for review, all functions performed shall be examined. For example, if a sub-unit... functions related to more than one of the areas. For example, coupon issuers must maintain a level of coupon...

  19. 7 CFR 275.7 - Selection of sub-units for review.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... situations, the sub-unit would be classified based upon its primary function exclusively. However, when any... classification and if selected for review, all functions performed shall be examined. For example, if a sub-unit... functions related to more than one of the areas. For example, coupon issuers must maintain a level of coupon...

  20. 7 CFR 275.7 - Selection of sub-units for review.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... situations, the sub-unit would be classified based upon its primary function exclusively. However, when any... classification and if selected for review, all functions performed shall be examined. For example, if a sub-unit... functions related to more than one of the areas. For example, coupon issuers must maintain a level of coupon...

  1. 7 CFR 275.7 - Selection of sub-units for review.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... situations, the sub-unit would be classified based upon its primary function exclusively. However, when any... classification and if selected for review, all functions performed shall be examined. For example, if a sub-unit... functions related to more than one of the areas. For example, coupon issuers must maintain a level of coupon...

  2. Aptamer modification improves the adenoviral transduction of malignant glioma cells.

    PubMed

    Chen, Hao; Zheng, Xiaojing; Di, BingYan; Wang, Dongyang; Zhang, Yaling; Xia, Haibin; Mao, Qinwen

    2013-12-01

    Adenovirus has shown increasing promise in the gene-viral therapy for glioblastoma, a treatment strategy that relies on the delivery of viruses or transgenes into tumor cells. However, targeting of adenovirus to human glioblastoma remains a challenge due to the low expression level of coxsackie and adenovirus receptor (CAR) in glioma cells. Aptamers are small and highly structured single-stranded oligonucleotides that bind at high affinity to a target molecule, and are good candidates for targeted imaging and therapy. In this study, to construct an aptamer-modified Ad5, we first genetically modified the HVR5 of Ad hexon by biotin acceptor peptide (BAP), which would be metabolically biotinylated during production in HEK293 cells, and then attached the biotin labeled aptamer to the modified Ad through avidin–biotin binding. The aptamers used in this study includes AS1411 and GBI-10. The former is a DNA aptamer that can bind to nucleolin, a nuclear matrix protein found on the surface of cancer cells. The latter is a DNA aptamer that can recognize the extracellular matrix protein tenascin-C on the surface of human glioblastoma cells. To examine if aptamer-modification of the hexon protein could improve the adenoviral transduction efficiency, a glioblastoma cell line, U251, was transduced with aptamer-modified Ads. The transduction efficiency of AS1411- or GBI-10-modified Ad was approximately 4.1-fold or 5.2-fold higher than that of the control. The data indicated that aptamer modified adenovirus would be a useful tool for cancer gene therapy. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

  3. Novel mechanism of JNK pathway activation by adenoviral E1A

    PubMed Central

    Morrison, Helen; Pospelova, Tatiana V.; Pospelov, Valery A.; Herrlich, Peter

    2014-01-01

    The adenoviral oncoprotein E1A influences cellular regulation by interacting with a number of cellular proteins. In collaboration with complementary oncogenes, E1A fully transforms primary cells. As part of this action, E1A inhibits transcription of c-Jun:Fos target genes while promoting that of c-Jun:ATF2-dependent genes including jun. Both c-Jun and ATF2 are hyperphosphorylated in response to E1A. In the current study, E1A was fused with the ligand binding domain of the estrogen receptor (E1A-ER) to monitor the immediate effect of E1A activation. With this approach we now show that E1A activates c-Jun N-terminal kinase (JNK), the upstream kinases MKK4 and MKK7, as well as the small GTPase Rac1. Activation of the JNK pathway requires the N-terminal domain of E1A, and, importantly, is independent of transcription. In addition, it requires the presence of ERM proteins. Downregulation of signaling components upstream of JNK inhibits E1A-dependent JNK/c-Jun activation. Taking these findings together, we show that E1A activates the JNK/c-Jun signaling pathway upstream of Rac1 in a transcription-independent manner, demonstrating a novel mechanism of E1A action. PMID:24742962

  4. Li{sub 2}Sr{sub 4}B{sub 12}O{sub 23}: A new alkali and alkaline-earth metal mixed borate with [B{sub 10}O{sub 18}]{sup 6-} network and isolated [B{sub 2}O{sub 5}]{sup 4-} unit

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

    Zhang Min; Graduate University of Chinese Academy of Sciences, Beijing 100049; Pan Shilie, E-mail: slpan@ms.xjb.ac.cn

    2012-06-15

    A novel ternary lithium strontium borate Li{sub 2}Sr{sub 4}B{sub 12}O{sub 23} crystal with size up to 20 mm Multiplication-Sign 10 mm Multiplication-Sign 4 mm has been grown via the top-seeded solution growth method below 730 Degree-Sign C. Single-crystal XRD analyses showed that Li{sub 2}Sr{sub 4}B{sub 12}O{sub 23} crystallizes in the monoclinic space group P2{sub 1}/c with a=6.4664(4) A, b=8.4878(4) A, c=15.3337(8) A, {beta}=102.02(3) Degree-Sign , Z=2. The crystal structure is composed of [B{sub 10}O{sub 18}]{sup 6-} network and isolated [B{sub 2}O{sub 5}]{sup 4-} unit. The IR spectrum further confirmed the presence of both BO{sub 3} and BO{sub 4} groups. TG-DSCmore » and Transmission spectrum were reported. Band structures and density of states were calculated. - Graphical abstract: A new phase, Li{sub 2}Sr{sub 4}B{sub 12}O{sub 23}, has been discovered in the ternary M{sub 2}O-M Prime O-B{sub 2}O{sub 3} (M=alkali-metal, M Prime =alkalineearth metal) system. The crystal structure consists of [B{sub 10}O{sub 18}]{sup 6-} network and isolated [B{sub 2}O{sub 5}]{sup 4-} unit. Highlights: Black-Right-Pointing-Pointer Li{sub 2}Sr{sub 4}B{sub 12}O{sub 23} is a a novel borate discovered in the M{sub 2}O-M Prime O-B{sub 2}O{sub 3} (M=alkali-metal, M Prime =alkaline-earth metal) system. Black-Right-Pointing-Pointer Li{sub 2}Sr{sub 4}B{sub 12}O{sub 23} crystal structure has a three-dimensional crystal structure with [B{sub 10}O{sub 18}]{sup 6-} network and isolated [B{sub 2}O{sub 5}]{sup 4-} unit. Black-Right-Pointing-Pointer Sr{sub 1} and Sr{sub 2} are located in two different channels constructed by {sup 3}{sub {infinity}}[B{sub 10}O{sub 18}] network.« less

  5. In vivo marking of spontaneous or vaccine-induced fibrosarcomas in the domestic house cat, using an adenoviral vector containing a bifunctional fusion protein, GAL-TEK.

    PubMed

    Marini, F C; Cannon, J P; Belmont, J W; Shillitoe, E J; Lapeyre, J N

    1995-09-01

    We evaluated the ability of a replication-deficient, recombinant adenoviral vector to transfer the bifunctional gene GAL-TEK, which expresses a marking/therapeutic gene product, to naturally occurring cat fibrosarcomas in situ. GAL-TEK contains an in-frame fusion of the bacterial LacZ gene for histochemical marking of tumors with beta-galactosidase (beta-Gal) and the HSV tk gene for enzyme-prodrug activation of the prodrug ganciclovir (GCV) to induce selective tumor cell killing. GAL-TEK bifunctional marking and cell killing activities were tested in vitro after adenoviral vector infection of HT1080 human fibrosarcoma cells. The tk activity of GAL-TEK is shown to be almost as potent as HSV tk to catalyze conversion of GCV to GCV nucleotides and promote selective cell killing. Using 8 cats with recurring 2.5-cm2 fibrosarcomas that either arose spontaneously or were induced by vaccine, we determined experimentally the administration routes and times required for optimum GAL-TEK gene transfer by beta-Gal histological staining and reverse transcriptase polymerase chain reaction to the multiple compartments of the growing fibrosarcomas consonant with minimizing collateral infection of neighboring tissues and other unwanted side effects.

  6. Improvement in adenoviral gene transfer efficiency after preincubation at +37 degrees C in vitro and in vivo.

    PubMed

    Kossila, Maija; Jauhiainen, Suvi; Laukkanen, Mikko O; Lehtolainen, Pauliina; Jääskeläinen, Maiju; Turunen, Päivi; Loimas, Sami; Wahlfors, Jarmo; Ylä-Herttuala, Seppo

    2002-01-01

    Adenovirus is a widely used vector in gene transfer experiments because it produces high transduction efficiency in vitro and in vivo by means of the coxsackie-adenovirus receptor (CAR) and major histocompatibility complex (MHC) class I alpha-2 domain. Adenoviral gene transfer efficiency has been reported to correlate with cellular CAR expression. We report here a simple method to increase adenoviral gene transfer efficiency in cells that do not express high levels of CAR: preincubation of adenovirus for 30-40 minutes at +37 degrees C significantly increased the transduction efficiency in vitro in CHO and BALB/3T3 cells, in which CAR is expressed at very low levels. Increased transduction efficiency of preincubated adenovirus was also detected in vivo in rat brain tissue. In addition, we found that adenoviruses were rapidly inactivated in human serum in a complement-independent manner, whereas fetal bovine serum (FBS) had hardly any effects on the viral infectivity. We conclude that preincubation of adenoviral vectors at +37 degrees C may substantially increase gene transfer efficiency in applications in which target cells do not express high levels of CAR.

  7. Development of an adenoviral vector with robust expression driven by p53

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

    Bajgelman, Marcio C.; Biotechnology Program, Biomedical Sciences Institute, University of Sao Paulo; Millennium Institute-Gene Therapy Network, Ministry of Science and Technology

    2008-02-05

    Here we introduce a new adenoviral vector where transgene expression is driven by p53. We first developed a synthetic promoter, referred to as PGTx{beta}, containing a p53-responsive element, a minimal promoter and the first intron of the rabbit {beta}-globin gene. Initial assays using plasmid-based vectors indicated that expression was tightly controlled by p53 and was 5-fold stronger than the constitutive CMV immediate early promoter/enhancer. The adenoviral vector, AdPG, was also shown to offer p53-responsive expression in prostate carcinoma cells LNCaP (wt p53), DU-145 (temperature sensitive mutant of p53) and PC3 (p53-null, but engineered to express temperature-sensitive p53 mutants). AdPG servedmore » as a sensor of p53 activity in LNCaP cells treated with chemotherapeutic agents. Since p53 can be induced by radiotherapy and chemotherapy, this new vector could be further developed for use in combination with conventional therapies to bring about cooperation between the genetic and pharmacologic treatment modalities.« less

  8. PCR diagnostics and monitoring of adenoviral infections in hematopoietic stem cell transplantation recipients

    PubMed Central

    Ussowicz, Marek; Rybka, Blanka; Wendycz-Domalewska, Danuta; Ryczan, Renata; Gorczyńska, Ewa; Kałwak, Krzysztof; Woźniak, Mieczysław

    2010-01-01

    After stem cell transplantation, human patients are prone to life-threatening opportunistic infections with a plethora of microorganisms. We report a retrospective study on 116 patients (98 children, 18 adults) who were transplanted in a pediatric bone marrow transplantation unit. Blood, urine and stool samples were collected and monitored for adenovirus (AdV) DNA using polymerase chain reaction (PCR) and real-time PCR (RT-PCR) on a regular basis. AdV DNA was detected in 52 (44.8%) patients, with mortality reaching 19% in this subgroup. Variables associated with adenovirus infection were transplantations from matched unrelated donors and older age of the recipient. An increased seasonal occurrence of adenoviral infections was observed in autumn and winter. Analysis of immune reconstitution showed a higher incidence of AdV infections during periods of low T-lymphocyte count. This study also showed a strong interaction between co-infections of AdV and BK polyomavirus in patients undergoing hematopoietic stem cell transplantations. PMID:20848295

  9. Ex Vivo Adenoviral Vector Gene Delivery Results in Decreased Vector-associated Inflammation Pre- and Post–lung Transplantation in the Pig

    PubMed Central

    Yeung, Jonathan C; Wagnetz, Dirk; Cypel, Marcelo; Rubacha, Matthew; Koike, Terumoto; Chun, Yi-Min; Hu, Jim; Waddell, Thomas K; Hwang, David M; Liu, Mingyao; Keshavjee, Shaf

    2012-01-01

    Acellular normothermic ex vivo lung perfusion (EVLP) is a novel method of donor lung preservation for transplantation. As cellular metabolism is preserved during perfusion, it represents a potential platform for effective gene transduction in donor lungs. We hypothesized that vector-associated inflammation would be reduced during ex vivo delivery due to isolation from the host immune system response. We compared ex vivo with in vivo intratracheal delivery of an E1-, E3-deleted adenoviral vector encoding either green fluorescent protein (GFP) or interleukin-10 (IL-10) to porcine lungs. Twelve hours after delivery, the lung was transplanted and the post-transplant function assessed. We identified significant transgene expression by 12 hours in both in vivo and ex vivo delivered groups. Lung function remained excellent in all ex vivo groups after viral vector delivery; however, as expected, lung function decreased in the in vivo delivered adenovirus vector encoding GFP (AdGFP) group with corresponding increases in IL-1β levels. Transplanted lung function was excellent in the ex vivo transduced lungs and inferior lung function was seen in the in vivo group after transplantation. In summary, ex vivo delivery of adenoviral gene therapy to the donor lung is superior to in vivo delivery in that it leads to less vector-associated inflammation and provides superior post-transplant lung function. PMID:22453765

  10. Guanine nucleotide regulatory protein co-purifies with the D/sub 2/-dopamine receptor

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

    Senogles, S.E.; Caron, M.G.

    1986-05-01

    The D/sub 2/-dopamine receptor from bovine anterior pituitary was purified approx.1000 fold by affinity chromatography on CMOS-Sepharose. Reconstitution of the affinity-purified receptor into phospholipid vesicles revealed the presence of high and low affinity agonist sites as detected by N-n-propylnorapomorphine (NPA) competition experiments with /sup 3/H-spiperone. High affinity agonist binding could be converted to the low affinity form by guanine nucleotides, indicating the presence of an endogenous guanine nucleotide binding protein (N protein) in the affinity-purified D/sub 2/ receptor preparations. Furthermore, this preparation contained an agonist-sensitive GTPase activity which was stimulated 2-3 fold over basal by 10 ..mu..M NPA. /sup 35/S-GTP..gamma..Smore » binding to these preparations revealed a stoichiometry of 0.4-0.7 mole N protein/mole receptor, suggesting the N protein may be specifically coupled with the purified D/sub 2/-dopamine receptor and not present as a contaminant. Pertussis toxin treatment of the affinity purified receptor preparations prevented high affinity agonist binding, as well as agonist stimulation of the GTPase activity, presumably by inactivating the associated N protein. Pertussis toxin lead to the ADP-ribosylation of a protein of 39-40K on SDS-PAGE. These findings indicate that an endogenous N protein, N/sub i/ or N/sub o/, co-purifies with the D/sub 2/-dopamine receptor which may reflect a precoupling of this receptor with an N protein within the membranes.« less

  11. Intra-testicular injection of adenoviral constructs results in Sertoli cell-specific gene expression and disruption of the seminiferous epithelium

    PubMed Central

    Hooley, R P; Paterson, M; Brown, P; Kerr, K; Saunders, P T K

    2009-01-01

    Spermatogenesis is a complex process that cannot be modelled in vitro. The somatic Sertoli cells (SCs) within the seminiferous tubules perform a key role in supporting maturation of germ cells (GCs). Progress has been made in determining what aspects of SC function are critical to maintenance of fertility by developing rodent models based on the Cre/LoxP system; however, this is time-consuming and is only applicable to mice. The aim of the present study was to establish methods for direct injection of adenoviral vectors containing shRNA constructs into the testis as a way of inducing target-selective knock-down in vivo. This paper describes a series of experiments using adenovirus expressing a green fluorescent protein (GFP) transgene. Injection via the efferent ductules resulted in SC-specific expression of GFP; expression levels paralleled the amount of infective viral particles injected. At the highest doses of virus seminiferous tubule architecture were grossly disturbed and immune cell invasion noted. At lower concentrations, the expression of GFP was variable/negligible, the seminiferous tubule lumen was maintained but stage-dependent GC loss and development of numerous basal vacuoles was observed. These resembled intercellular dilations of SC junctional complexes previously described in rats and may be a consequence of disturbances in SC function due to interaction of the viral particles with the coxsackie/adenovirus receptor that is a component of the junctional complexes within the blood testis barrier. In conclusion, intra-testicular injection of adenoviral vectors disturbs SC function in vivo and future work will therefore focus on the use of lentiviral delivery systems. PMID:18955374

  12. Reversal of experimental colitis disease activity in mice following administration of an adenoviral IL-10 vector.

    PubMed

    Sasaki, Makoto; Mathis, J Michael; Jennings, Merilyn H; Jordan, Paul; Wang, Yuping; Ando, Tomoaki; Joh, Takashi; Alexander, J Steven

    2005-10-31

    Genetic deficiency in the expression of interleukin-10 (IL-10) is associated with the onset and progression of experimental inflammatory bowel disease (IBD). The clinical significance of IL-10 expression is supported by studies showing that immune-augmentation of IL-10 prevents inflammation and mucosal damage in animal models of colitis and in human colitis. Interleukin-10 (IL-10), an endogenous anti-inflammatory and immunomodulating cytokine, has been shown to prevent some inflammation and injury in animal and clinical studies, but the efficacy of IL-10 treatment remains unsatisfactory. We found that intra-peritoneal administration of adenoviral IL-10 to mice significantly reversed colitis induced by administration of 3% DSS (dextran sulfate), a common model of colitis. Adenoviral IL-10 (Ad-IL10) transfected mice developed high levels of IL-10 (394 +/- 136 pg/ml) within the peritoneal cavity where the adenovirus was expressed. Importantly, when given on day 4 (after the induction of colitis w/DSS), Ad-IL10 significantly reduced disease activity and weight loss and completely prevented histopathologic injury to the colon at day 10. Mechanistically, compared to Ad-null and DSS treated mice, Ad-IL10 and DSS-treated mice were able to suppress the expression of MAdCAM-1, an endothelial adhesion molecule associated with IBD. Our results suggest that Ad-IL10 (adenoviral IL-10) gene therapy of the intestine or peritoneum may be useful in the clinical treatment of IBD, since we demonstrated that this vector can reverse the course of an existing gut inflammation and markers of inflammation.

  13. Reversal of experimental colitis disease activity in mice following administration of an adenoviral IL-10 vector

    PubMed Central

    Sasaki, Makoto; Mathis, J Michael; Jennings, Merilyn H; Jordan, Paul; Wang, Yuping; Ando, Tomoaki; Joh, Takashi; Alexander, J Steven

    2005-01-01

    Genetic deficiency in the expression of interleukin-10 (IL-10) is associated with the onset and progression of experimental inflammatory bowel disease (IBD). The clinical significance of IL-10 expression is supported by studies showing that immune-augmentation of IL-10 prevents inflammation and mucosal damage in animal models of colitis and in human colitis. Interleukin-10 (IL-10), an endogenous anti-inflammatory and immunomodulating cytokine, has been shown to prevent some inflammation and injury in animal and clinical studies, but the efficacy of IL-10 treatment remains unsatisfactory. We found that intra-peritoneal administration of adenoviral IL-10 to mice significantly reversed colitis induced by administration of 3% DSS (dextran sulfate), a common model of colitis. Adenoviral IL-10 (Ad-IL10) transfected mice developed high levels of IL-10 (394 +/- 136 pg/ml) within the peritoneal cavity where the adenovirus was expressed. Importantly, when given on day 4 (after the induction of colitis w/DSS), Ad-IL10 significantly reduced disease activity and weight loss and completely prevented histopathologic injury to the colon at day 10. Mechanistically, compared to Ad-null and DSS treated mice, Ad-IL10 and DSS-treated mice were able to suppress the expression of MAdCAM-1, an endothelial adhesion molecule associated with IBD. Our results suggest that Ad-IL10 (adenoviral IL-10) gene therapy of the intestine or peritoneum may be useful in the clinical treatment of IBD, since we demonstrated that this vector can reverse the course of an existing gut inflammation and markers of inflammation. PMID:16259632

  14. Adenoviral vector tethering to metal surfaces via hydrolysable cross-linkers for the modulation of vector release and transduction

    PubMed Central

    Fishbein, Ilia; Forbes, Scott P.; Chorny, Michael; Connolly, Jeanne M.; Adamo, Richard F.; Corrales, Ricardo; Alferiev, Ivan S.; Levy, Robert J.

    2013-01-01

    The use of arterial stents and other medical implants as a delivery platform for surface immobilized gene vectors allows for safe and efficient localized expression of therapeutic transgenes. In this study we investigate the use of hydrolysable cross-linkers with distinct kinetics of hydrolysis for delivery of gene vectors from polyallylamine bisphosphonate-modified metal surfaces. Three cross-linkers with the estimated t1/2 of ester bonds hydrolysis of 5, 12 and 50 days demonstrated a cumulative 20%, 39% and 45% vector release, respectively, after 30 days exposure to physiological buffer at 37°C. Transgene expression in endothelial and smooth muscles cells transduced with substrate immobilized adenovirus resulted in significantly different expression profiles for each individual cross-linker. Furthermore, immobilization of adenoviral vectors effectively extended their transduction effectiveness beyond the initial phase of release. Transgene expression driven by adenovirus-tethered stents in rat carotid arteries demonstrated that a faster rate of cross-linker hydrolysis resulted in higher expression levels at day 1, which declined by day 8 after stent implantation, while inversely, slower hydrolysis was associated with increased arterial expression at day 8 in comparison with day 1. In conclusion, adjustable release of transduction-competent adenoviral vectors from metallic surfaces can be achieved, both in vitro and in vivo, through surface immobilization of adenoviral vectors using hydrolysable cross-linkers with structure-specific release kinetics. PMID:23777912

  15. Upgrading HepG2 cells with adenoviral vectors that encode drug-metabolizing enzymes: application for drug hepatotoxicity testing.

    PubMed

    Gómez-Lechón, M José; Tolosa, Laia; Donato, M Teresa

    2017-02-01

    Drug attrition rates due to hepatotoxicity are an important safety issue considered in drug development. The HepG2 hepatoma cell line is currently being used for drug-induced hepatotoxicity evaluations, but its expression of drug-metabolizing enzymes is poor compared with hepatocytes. Different approaches have been proposed to upgrade HepG2 cells for more reliable drug-induced liver injury predictions. Areas covered: We describe the advantages and limitations of HepG2 cells transduced with adenoviral vectors that encode drug-metabolizing enzymes for safety risk assessments of bioactivable compounds. Adenoviral transduction facilitates efficient and controlled delivery of multiple drug-metabolizing activities to HepG2 cells at comparable levels to primary human hepatocytes by generating an 'artificial hepatocyte'. Furthermore, adenoviral transduction enables the design of tailored cells expressing particular metabolic capacities. Expert opinion: Upgraded HepG2 cells that recreate known inter-individual variations in hepatic CYP and conjugating activities due to both genetic (e.g., polymorphisms) or environmental (e.g., induction, inhibition) factors seems a suitable model to identify bioactivable drug and conduct hepatotoxicity risk assessments. This strategy should enable the generation of customized cells by reproducing human pheno- and genotypic CYP variability to represent a valuable human hepatic cell model to develop new safer drugs and to improve existing predictive toxicity assays.

  16. Alanine–glyoxylate aminotransferase-deficient mice, a model for primary hyperoxaluria that responds to adenoviral gene transfer

    PubMed Central

    Salido, Eduardo C.; Li, Xiao M.; Lu, Yang; Wang, Xia; Santana, Alfredo; Roy-Chowdhury, Namita; Torres, Armando; Shapiro, Larry J.; Roy-Chowdhury, Jayanta

    2006-01-01

    Mutations in the alanine–glyoxylate amino transferase gene (AGXT) are responsible for primary hyperoxaluria type I, a rare disease characterized by excessive hepatic oxalate production that leads to renal failure. We generated a null mutant mouse by targeted mutagenesis of the homologous gene, Agxt, in embryonic stem cells. Mutant mice developed normally, and they exhibited hyperoxaluria and crystalluria. Approximately half of the male mice in mixed genetic background developed calcium oxalate urinary stones. Severe nephrocalcinosis and renal failure developed after enhancement of oxalate production by ethylene glycol administration. Hepatic expression of human AGT1, the protein encoded by AGXT, by adenoviral vector-mediated gene transfer in Agxt−/− mice normalized urinary oxalate excretion and prevented oxalate crystalluria. Subcellular fractionation and immunofluorescence studies revealed that, as in the human liver, the expressed wild-type human AGT1 was predominantly localized in mouse hepatocellular peroxisomes, whereas the most common mutant form of AGT1 (G170R) was localized predominantly in the mitochondria. PMID:17110443

  17. Pulsed Electron Beam Water Radiolysis for Sub-Microsecond Hydroxyl Radical Protein Footprinting

    PubMed Central

    Watson, Caroline; Janik, Ireneusz; Zhuang, Tiandi; Charvátová, Olga; Woods, Robert J.; Sharp, Joshua S.

    2009-01-01

    Hydroxyl radical footprinting is a valuable technique for studying protein structure, but care must be taken to ensure that the protein does not unfold during the labeling process due to oxidative damage. Footprinting methods based on sub-microsecond laser photolysis of peroxide that complete the labeling process faster than the protein can unfold have been recently described; however, the mere presence of large amounts of hydrogen peroxide can also cause uncontrolled oxidation and minor conformational changes. We have developed a novel method for sub-microsecond hydroxyl radical protein footprinting using a pulsed electron beam from a 2 MeV Van de Graaff electron accelerator to generate a high concentration of hydroxyl radicals by radiolysis of water. The amount of oxidation can be controlled by buffer composition, pulsewidth, dose, and dissolved nitrous oxide gas in the sample. Our results with ubiquitin and β-lactoglobulin A demonstrate that one sub-microsecond electron beam pulse produces extensive protein surface modifications. Highly reactive residues that are buried within the protein structure are not oxidized, indicating that the protein retains its folded structure during the labeling process. Time-resolved spectroscopy indicates that the major part of protein oxidation is complete in a timescale shorter than that of large scale protein motions. PMID:19265387

  18. Smallest fullerene-like silicon cage stabilized by a V{sub 2} unit

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

    Xu, Hong-Guang, E-mail: xuhong@iccas.ac.cn, E-mail: zhengwj@iccas.ac.cn; Kong, Xiang-Yu; Deng, Xiao-Jiao

    We conducted a combined anion photoelectron spectroscopy and density functional theory study on V{sub 2}Si{sub 20} cluster. Our results show that the V{sub 2}Si{sub 20} cluster has an elongated dodecahedron cage structure with a V{sub 2} unit encapsulated inside the cage. It is the smallest fullerene-like silicon cage and can be used as building block to make cluster-assembled materials, such as pearl-chain style nanowires.

  19. Α2 GABAA receptor sub-units in the ventral hippocampus and α5 GABAA receptor sub-units in the dorsal hippocampus mediate anxiety and fear memory.

    PubMed

    McEown, K; Treit, D

    2013-11-12

    Temporary neuronal inactivation of the ventral hippocampus with the GABAA agonist muscimol suppresses unconditioned fear behavior (anxiety) but inactivation of the dorsal hippocampus does not. On the other hand, inactivating the dorsal hippocampus disrupts fear memory, while inactivating the ventral hippocampus does not. Here we investigate the roles of hippocampal GABAA receptor sub-units in mediating these anxiolytic and amnesic effects of GABAA receptor agonists. We microinfused TPA023 (α2 agonist) or TB-21007 (inverse α5 agonist) into the dorsal or ventral hippocampus prior to testing rats in two animal models of anxiety: the elevated plus-maze and shock-probe burying test. Twenty-four hours later rats were re-tested in the shock-probe chamber with a non-electrified probe to assess their memory of the initial shock-probe experience (i.e., fear memory). We found that TPA023 was anxiolytic in the plus-maze and shock-probe burying tests when microinfused into the ventral hippocampus. However, TPA023 did not affect anxiety-related behavior when infused into the dorsal hippocampus. Conversely, we found that the α5 sub-unit inverse agonist TB-21007 impaired rats' memory of the initial shock-probe experience when infused into the dorsal hippocampus, but not when infused into the ventral hippocampus. This double dissociation suggests that α2 GABAA receptor sub-units in the ventral hippocampus mediate unconditioned fear or anxiety, while α5 GABAA receptor sub-units in the dorsal hippocampus mediate conditioned fear memory. Copyright © 2013 IBRO. Published by Elsevier Ltd. All rights reserved.

  20. Determining the sub-cellular localization of proteins within Caenorhabditis elegans body wall muscle.

    PubMed

    Meissner, Barbara; Rogalski, Teresa; Viveiros, Ryan; Warner, Adam; Plastino, Lorena; Lorch, Adam; Granger, Laure; Segalat, Laurent; Moerman, Donald G

    2011-01-01

    Determining the sub-cellular localization of a protein within a cell is often an essential step towards understanding its function. In Caenorhabditis elegans, the relatively large size of the body wall muscle cells and the exquisite organization of their sarcomeres offer an opportunity to identify the precise position of proteins within cell substructures. Our goal in this study is to generate a comprehensive "localizome" for C. elegans body wall muscle by GFP-tagging proteins expressed in muscle and determining their location within the cell. For this project, we focused on proteins that we know are expressed in muscle and are orthologs or at least homologs of human proteins. To date we have analyzed the expression of about 227 GFP-tagged proteins that show localized expression in the body wall muscle of this nematode (e.g. dense bodies, M-lines, myofilaments, mitochondria, cell membrane, nucleus or nucleolus). For most proteins analyzed in this study no prior data on sub-cellular localization was available. In addition to discrete sub-cellular localization we observe overlapping patterns of localization including the presence of a protein in the dense body and the nucleus, or the dense body and the M-lines. In total we discern more than 14 sub-cellular localization patterns within nematode body wall muscle. The localization of this large set of proteins within a muscle cell will serve as an invaluable resource in our investigation of muscle sarcomere assembly and function.

  1. Observation of gold sub-nanocluster nucleation within a crystalline protein cage

    NASA Astrophysics Data System (ADS)

    Maity, Basudev; Abe, Satoshi; Ueno, Takafumi

    2017-03-01

    Protein scaffolds provide unique metal coordination environments that promote biomineralization processes. It is expected that protein scaffolds can be developed to prepare inorganic nanomaterials with important biomedical and material applications. Despite many promising applications, it remains challenging to elucidate the detailed mechanisms of formation of metal nanoparticles in protein environments. In the present work, we describe a crystalline protein cage constructed by crosslinking treatment of a single crystal of apo-ferritin for structural characterization of the formation of sub-nanocluster with reduction reaction. The crystal structure analysis shows the gradual movement of the Au ions towards the centre of the three-fold symmetric channels of the protein cage to form a sub-nanocluster with accompanying significant conformational changes of the amino-acid residues bound to Au ions during the process. These results contribute to our understanding of metal core formation as well as interactions of the metal core with the protein environment.

  2. Protein accumulation in aleurone cells, sub-aleurone cells and the center starch endosperm of cereals.

    PubMed

    Zheng, Yankun; Wang, Zhong

    2014-10-01

    There are mainly three endosperm storage tissues in the cereal endosperm: aleurone cells, sub-aleurone cells and the center starch endosperm. The protein accumulation is very different in the three endosperm storage tissues. The aleurone cells accumulate protein in aleurone granules. The sub-aleurone cells and the center starch endosperm accumulate protein in endoplasmic reticulum-derived protein bodies and vacuolar protein bodies. Proteins are deposited in different patterns within different endosperm storage tissues probably because of the special storage properties of these tissues. There are several special genes and other molecular factors to mediate the protein accumulation in these tissues. Different proteins have distinct functions in the protein body formation and the protein interactions determine protein body assembly. There are both cooperation and competition relationships between protein, starch and lipid in the cereal endosperm. This paper reviews the latest investigations on protein accumulation in aleurone cells, sub-aleurone cells and the center starch endosperm. Useful information will be supplied for future investigations on the cereal endosperm development.

  3. Surmounting limited gene delivery into primary immune cell populations: Efficient cell type-specific adenoviral transduction by CAR.

    PubMed

    Clausen, Björn E; Brand, Anna; Karram, Khalad

    2015-06-01

    Ectopic gene expression studies in primary immune cells have been notoriously difficult to perform due to the limitations in conventional transfection and viral transduction methods. Although replication-defective adenoviruses provide an attractive alternative for gene delivery, their use has been hampered by the limited susceptibility of murine leukocytes to adenoviral infection, due to insufficient expression of the human coxsackie/adenovirus receptor (CAR). In this issue of the European Journal of Immunology, Heger et al. [Eur. J. Immunol. 2015. 45: XXXX-XXXX] report the generation of transgenic mice that enable conditional Cre/loxP-mediated expression of human CAR. The authors demonstrate that this R26/CAG-CAR∆1(StopF) mouse strain facilitates the faithful monitoring of Cre activity in situ as well as the specific and efficient adenoviral transduction of primary immune cell populations in vitro. Further tweaking of the system towards more efficient gene transfer in vivo remains a future challenge. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. A World History Sub-Unit: Teaching about Turkey.

    ERIC Educational Resources Information Center

    Lynn, Karen

    This document is a sub-unit teaching plan for world history teachers who want to use multicultural concepts in the world history curriculum. The objective explored includes a student response to the Turkish question of "Who are we"? Teacher preparation involves defining social and cultural roots and outlining periods of Turkish history.…

  5. Highly efficient gene inactivation by adenoviral CRISPR/Cas9 in human primary cells

    PubMed Central

    Tielen, Frans; Elstak, Edo; Benschop, Julian; Grimbergen, Max; Stallen, Jan; Janssen, Richard; van Marle, Andre; Essrich, Christian

    2017-01-01

    Phenotypic assays using human primary cells are highly valuable tools for target discovery and validation in drug discovery. Expression knockdown (KD) of such targets in these assays allows the investigation of their role in models of disease processes. Therefore, efficient and fast modes of protein KD in phenotypic assays are required. The CRISPR/Cas9 system has been shown to be a versatile and efficient means of gene inactivation in immortalized cell lines. Here we describe the use of adenoviral (AdV) CRISPR/Cas9 vectors for efficient gene inactivation in two human primary cell types, normal human lung fibroblasts and human bronchial epithelial cells. The effects of gene inactivation were studied in the TGF-β-induced fibroblast to myofibroblast transition assay (FMT) and the epithelial to mesenchymal transition assay (EMT), which are SMAD3 dependent and reflect pathogenic mechanisms observed in fibrosis. Co-transduction (co-TD) of AdV Cas9 with SMAD3-targeting guide RNAs (gRNAs) resulted in fast and efficient genome editing judged by insertion/deletion (indel) formation, as well as significant reduction of SMAD3 protein expression and nuclear translocation. This led to phenotypic changes downstream of SMAD3 inhibition, including substantially decreased alpha smooth muscle actin and fibronectin 1 expression, which are markers for FMT and EMT, respectively. A direct comparison between co-TD of separate Cas9 and gRNA AdV, versus TD with a single “all-in-one” Cas9/gRNA AdV, revealed that both methods achieve similar levels of indel formation. These data demonstrate that AdV CRISPR/Cas9 is a useful and efficient tool for protein KD in human primary cell phenotypic assays. The use of AdV CRISPR/Cas9 may offer significant advantages over the current existing tools and should enhance target discovery and validation opportunities. PMID:28800587

  6. Statistical discovery of site inter-dependencies in sub-molecular hierarchical protein structuring

    PubMed Central

    2012-01-01

    Background Much progress has been made in understanding the 3D structure of proteins using methods such as NMR and X-ray crystallography. The resulting 3D structures are extremely informative, but do not always reveal which sites and residues within the structure are of special importance. Recently, there are indications that multiple-residue, sub-domain structural relationships within the larger 3D consensus structure of a protein can be inferred from the analysis of the multiple sequence alignment data of a protein family. These intra-dependent clusters of associated sites are used to indicate hierarchical inter-residue relationships within the 3D structure. To reveal the patterns of associations among individual amino acids or sub-domain components within the structure, we apply a k-modes attribute (aligned site) clustering algorithm to the ubiquitin and transthyretin families in order to discover associations among groups of sites within the multiple sequence alignment. We then observe what these associations imply within the 3D structure of these two protein families. Results The k-modes site clustering algorithm we developed maximizes the intra-group interdependencies based on a normalized mutual information measure. The clusters formed correspond to sub-structural components or binding and interface locations. Applying this data-directed method to the ubiquitin and transthyretin protein family multiple sequence alignments as a test bed, we located numerous interesting associations of interdependent sites. These clusters were then arranged into cluster tree diagrams which revealed four structural sub-domains within the single domain structure of ubiquitin and a single large sub-domain within transthyretin associated with the interface among transthyretin monomers. In addition, several clusters of mutually interdependent sites were discovered for each protein family, each of which appear to play an important role in the molecular structure and/or function

  7. Statistical discovery of site inter-dependencies in sub-molecular hierarchical protein structuring.

    PubMed

    Durston, Kirk K; Chiu, David Ky; Wong, Andrew Kc; Li, Gary Cl

    2012-07-13

    Much progress has been made in understanding the 3D structure of proteins using methods such as NMR and X-ray crystallography. The resulting 3D structures are extremely informative, but do not always reveal which sites and residues within the structure are of special importance. Recently, there are indications that multiple-residue, sub-domain structural relationships within the larger 3D consensus structure of a protein can be inferred from the analysis of the multiple sequence alignment data of a protein family. These intra-dependent clusters of associated sites are used to indicate hierarchical inter-residue relationships within the 3D structure. To reveal the patterns of associations among individual amino acids or sub-domain components within the structure, we apply a k-modes attribute (aligned site) clustering algorithm to the ubiquitin and transthyretin families in order to discover associations among groups of sites within the multiple sequence alignment. We then observe what these associations imply within the 3D structure of these two protein families. The k-modes site clustering algorithm we developed maximizes the intra-group interdependencies based on a normalized mutual information measure. The clusters formed correspond to sub-structural components or binding and interface locations. Applying this data-directed method to the ubiquitin and transthyretin protein family multiple sequence alignments as a test bed, we located numerous interesting associations of interdependent sites. These clusters were then arranged into cluster tree diagrams which revealed four structural sub-domains within the single domain structure of ubiquitin and a single large sub-domain within transthyretin associated with the interface among transthyretin monomers. In addition, several clusters of mutually interdependent sites were discovered for each protein family, each of which appear to play an important role in the molecular structure and/or function. Our results

  8. Clinical and Antiviral Efficacy of an Ophthalmic Formulation of Dexamethasone Povidone-Iodine in a Rabbit Model of Adenoviral Keratoconjunctivitis

    PubMed Central

    Clement, Christian; Capriotti, Joseph A.; Kumar, Manish; Hobden, Jeffery A.; Foster, Timothy P.; Bhattacharjee, Partha S.; Thompson, Hilary W.; Mahmud, Rashed; Liang, Bo

    2011-01-01

    Purpose. To determine the efficacy of a new formulation of topical dexamethasone 0.1%/povidone-iodine 0.4% (FST-100) in reducing clinical symptoms and infectious viral titers in a rabbit model of adenoviral keratoconjunctivitis. Methods. Rabbit corneas were inoculated bilaterally with 2 × 106 plaque-forming-units (PFU) of adenovirus type 5 (Ad5) after corneal scarification. Animals were randomized 1:1:1:1 (five rabbits per group) to FST-100, 0.5% cidofovir, tobramycin/dexamethasone (Tobradex; Alcon Laboratories, Fort Worth, TX) ophthalmic suspension, and balanced salt solution (BSS; Alcon Laboratories). Treatment began 12 hours after viral inoculation and continued for 7 consecutive days. The eyes were clinically scored daily for scleral inflammation (injection), ocular neovascularization, eyelid inflammation (redness), friability of vasculature, inflammatory discharge (pus), and epiphora (excessive tearing). Eye swabs were collected daily before treatment for the duration of the study. Virus was eluted from the swabs and PFU determined by titration on human A549 cells, according to standard procedures. Results. The FST-100 treatment resulted in significantly lower clinical scores (P < 0.05) than did the other treatments. The 0.5% cidofovir exhibited the most ocular toxicity compared with FST-100, tobramycin/dexamethasone, and balanced salt solution treatments. FST-100 and 0.5% cidofovir significantly (P < 0.05) reduced viral titers compared with tobramycin/dexamethasone or balanced salt solution. Conclusions. FST-100 was the most efficacious in minimizing the clinical symptoms of adenovirus infection in rabbit eyes. FST-100 and 0.5% cidofovir were both equally effective in reducing viral titers and decreasing the duration of viral shedding. By providing symptomatic relief in addition to reducing infectious virus titers, FST-100 should be a valuable addition to treatment of epidemic adenoviral keratoconjunctivitis. PMID:20702820

  9. LY294002 enhances expression of proteins encoded by recombinant replication-defective adenoviruses via mTOR- and non-mTOR-dependent mechanisms.

    PubMed

    Shepelev, Mikhail V; Korobko, Elena V; Vinogradova, Tatiana V; Kopantsev, Eugene P; Korobko, Igor V

    2013-03-04

    Adenovirus-based drugs are efficient when combined with other anticancer treatments. Here we show that treatment with LY294002 and LY303511 upregulates expression of recombinant proteins encoded by replication-defective adenoviruses, including expression of therapeutically valuable combination of herpes simplex virus thymidine kinase controlled by human telomerase reverse transcriptase promoter (Ad-hTERT-HSVtk). In line with this, treatment with LY294002 synergized with Ad-hTERT-HSVtk infection in the presence of gancyclovir prodrug on Calu-I lung cancer cell death. The effect of LY294002 and LY303511 on adenovirus-delivered transgene expression was demonstrated in 4 human lung cancer cell lines. LY294002-induced upregulation of adenovirally delivered transgene is mediated in part by direct inhibition of mTOR protein kinase in mTORC2 signaling complex thus suggesting that anticancer drugs targeting mTOR will also enhance expression of transgenes delivered with adenoviral vectors. As both LY294002 and LY303511 are candidate prototypic anticancer drugs, and many mTOR inhibitors for cancer treatment are under development, our results have important implication for development of future therapeutic strategies with adenoviral gene delivery.

  10. A Genetically Modified Adenoviral Vector with a Phage Display-Derived Peptide Incorporated into Fiber Fibritin Chimera Prolongs Survival in Experimental Glioma.

    PubMed

    Kim, Julius W; Kane, J Robert; Young, Jacob S; Chang, Alan L; Kanojia, Deepak; Morshed, Ramin A; Miska, Jason; Ahmed, Atique U; Balyasnikova, Irina V; Han, Yu; Zhang, Lingjiao; Curiel, David T; Lesniak, Maciej S

    2015-09-01

    The dismal clinical context of advanced-grade glioma demands the development of novel therapeutic strategies with direct patient impact. Adenovirus-mediated virotherapy represents a potentially effective approach for glioma therapy. In this research, we generated a novel glioma-specific adenovirus by instituting more advanced genetic modifications that can maximize the efficiency and safety of therapeutic adenoviral vectors. In this regard, a glioma-specific targeted fiber was developed through the incorporation of previously published glioma-specific, phage-panned peptide (VWT peptide) on a fiber fibritin-based chimeric fiber, designated as "GliomaFF." We showed that the entry of this virus was highly restricted to glioma cells, supporting the specificity imparted by the phage-panned peptide. In addition, the stability of the targeting moiety presented by fiber fibritin structure permitted greatly enhanced infectivity. Furthermore, the replication of this virus was restricted in glioma cells by controlling expression of the E1 gene under the activity of the tumor-specific survivin promoter. Using this approach, we were able to explore the combinatorial efficacy of various adenoviral modifications that could amplify the specificity, infectivity, and exclusive replication of this therapeutic adenovirus in glioma. Finally, virotherapy with this modified virus resulted in up to 70% extended survival in an in vivo murine glioma model. These data demonstrate that this novel adenoviral vector is a safe and efficient treatment for this difficult malignancy.

  11. Adenoviral transfer of the heme oxygenase-1 gene protects striatal astrocytes from heme-mediated oxidative injury.

    PubMed

    Teng, Zhi-Ping; Chen, Jing; Chau, Lee-Young; Galunic, Nicholas; Regan, Raymond F

    2004-11-01

    Heme oxygenase-1 (HO-1) is induced in the CNS after hemorrhage, and may have an effect on injury to surrounding tissue. Hemin, the preferred substrate of HO, is a neurotoxin that is present in intracranial hematomas. In a prior study, we observed that HO inhibitors increased the vulnerability of cultured cortical astrocytes to heme-mediated oxidative injury. To investigate the effect of HO more specifically, we used an adenoviral vector encoding the human HO-1 gene to specifically increase HO-1 expression. Incubation with 100 MOI of the HO-1 adenovirus (Adv-HHO-1) for 24 h increased both HO-1 protein and HO activity; a control adenovirus lacking the HO-1 gene had no effect. Using a DNA probe that was specific for human HO-1, 80.5 +/- 7.2% of astrocytes were observed to be infected by in situ hybridization. The cell death produced by 30-60 microM hemin was significantly reduced by pretreatment with 100 MOI Adv-HHO-1, as assessed by LDH release, propidium iodide exclusion, and MTT reduction assay. The threefold increase in cell protein oxidation produced by hemin was also attenuated in cultures pretreated with Adv-HHO-1. These results support the hypothesis that HO-1 protects astrocytes from heme-mediated oxidative injury. Specifically increasing astrocytic HO-1 by gene transfer may have a beneficial effect on hemorrhagic CNS injury.

  12. Chimpanzee adenoviral vectors as vaccines for outbreak pathogens

    PubMed Central

    2017-01-01

    ABSTRACT The 2014–15 Ebola outbreak in West Africa highlighted the potential for large disease outbreaks caused by emerging pathogens and has generated considerable focus on preparedness for future epidemics. Here we discuss drivers, strategies and practical considerations for developing vaccines against outbreak pathogens. Chimpanzee adenoviral (ChAd) vectors have been developed as vaccine candidates for multiple infectious diseases and prostate cancer. ChAd vectors are safe and induce antigen-specific cellular and humoral immunity in all age groups, as well as circumventing the problem of pre-existing immunity encountered with human Ad vectors. For these reasons, such viral vectors provide an attractive platform for stockpiling vaccines for emergency deployment in response to a threatened outbreak of an emerging pathogen. Work is already underway to develop vaccines against a number of other outbreak pathogens and we will also review progress on these approaches here, particularly for Lassa fever, Nipah and MERS. PMID:29083948

  13. Adenoviral Vector Immunity: Its Implications and circumvention strategies

    PubMed Central

    Ahi, Yadvinder S.; Bangari, Dinesh S.; Mittal, Suresh K.

    2014-01-01

    Adenoviral (Ad) vectors have emerged as a promising gene delivery platform for a variety of therapeutic and vaccine purposes during last two decades. However, the presence of preexisting Ad immunity and the rapid development of Ad vector immunity still pose significant challenges to the clinical use of these vectors. Innate inflammatory response following Ad vector administration may lead to systemic toxicity, drastically limit vector transduction efficiency and significantly abbreviate the duration of transgene expression. Currently, a number of approaches are being extensively pursued to overcome these drawbacks by strategies that target either the host or the Ad vector. In addition, significant progress has been made in the development of novel Ad vectors based on less prevalent human Ad serotypes and nonhuman Ad. This review provides an update on our current understanding of immune responses to Ad vectors and delineates various approaches for eluding Ad vector immunity. Approaches targeting the host and those targeting the vector are discussed in light of their promises and limitations. PMID:21453277

  14. The sodium pump α1 sub-unit: a disease progression–related target for metastatic melanoma treatment

    PubMed Central

    Mathieu, Véronique; Pirker, Christine; Martin de Lassalle, Elisabeth; Vernier, Mathieu; Mijatovic, Tatjana; DeNeve, Nancy; Gaussin, Jean-François; Dehoux, Mischael; Lefranc, Florence; Berger, Walter; Kiss, Robert

    2009-01-01

    Melanomas remain associated with dismal prognosis because they are naturally resistant to apoptosis and they markedly metastasize. Up-regulated expression of sodium pump α sub-units has previously been demonstrated when comparing metastatic to non-metastatic melanomas. Our previous data revealed that impairing sodium pump α1 activity by means of selective ligands, that are cardiotonic steroids, markedly impairs cell migration and kills apoptosis-resistant cancer cells. The objective of this study was to determine the expression levels of sodium pump α sub-units in melanoma clinical samples and cell lines and also to characterize the role of α1 sub-units in melanoma cell biology. Quantitative RT-PCR, Western blotting and immunohistochemistry were used to determine the expression levels of sodium pump α sub-units. In vitro cytotoxicity of various cardenolides and of an anti-α1 siRNA was evaluated by means of MTT assay, quantitative videomicroscopy and through apoptosis assays. The in vivo activity of a novel cardenolide UNBS1450 was evaluated in a melanoma brain metastasis model. Our data show that all investigated human melanoma cell lines expressed high levels of the α1 sub-unit, and 33% of human melanomas displayed significant α1 sub-unit expression in correlation with the Breslow index. Furthermore, cardenolides (notably UNBS1450; currently in Phase I clinical trials) displayed marked anti-tumour effects against melanomas in vitro. This activity was closely paralleled by decreases in cMyc expression and by increases in apoptotic features. UNBS1450 also displayed marked anti-tumour activity in the aggressive human metastatic brain melanoma model in vivo. The α1 sodium pump sub-unit could represent a potential novel target for combating melanoma. PMID:19243476

  15. Adenoviral Mediated Gene Transfer of IGF-1 Enhances Wound Healing and Induces Angiogenesis

    PubMed Central

    Balaji, S.; LeSaint, M.; Bhattacharya, S. S.; Moles, C.; Dhamija, Y.; Kidd, M.; Le, L.D.; King, A.; Shaaban, A.; Crombleholme, T. M.; Bollyky, P.; Keswani, S. G.

    2014-01-01

    Background Chronic wounds are characterized by a wound healing and neovascularization deficit. Strategies to increase neovascularization can significantly improve chronic wound healing. Insulin like growth factor (IGF-1) is reported to be a keratinocyte mitogen and is believed to induce angiogenesis via a vascular endothelial growth factor (VEGF) dependent pathway. Using a novel ex vivo human dermal wound model and a diabetic impaired wound healing murine model, we hypothesized that adenoviral over expression of IGF-1 (Ad-IGF-1) will enhance wound healing and induce angiogenesis through a VEGF dependent pathway. Methods Ex vivo: 6 mm full thickness punch biopsies were obtained from normal human skin, and 3 mm full thickness wounds were created at the center. Skin explants were maintained at air liquid interface. Db/db murine model: 8 mm full thickness dorsal wounds in diabetic (db/db) mice were created. Treatment groups in both human ex vivo and in vivo db/db wound models include 1×108 PFU of Ad-IGF-1 or Ad-LacZ, and PBS (n=4–5/group). Cytotoxicity (LDH) was quantified at days 3, 5 and 7 for the human ex vivo wound model. Epithelial gap closure (H&E; Trichrome), VEGF expression (ELISA) and capillary density (CD 31+ CAPS/HPF) were analyzed at day 7. Results In the human ex vivo organ culture, the adenoviral vectors did not demonstrate any significant difference in cytotoxicity compared to PBS. Ad-IGF-1 over expression significantly increases basal keratinocyte migration, with no significant effect on epithelial gap closure. There was a significant increase in capillary density in the Ad-IGF-1 wounds. However, there was no effect on VEGF levels in Ad-IGF-1 samples compared to controls. In db/db wounds, Ad-IGF-1 over expression significantly improves epithelial gap closure and granulation tissue with a dense cellular infiltrate compared to controls. Ad-IGF-1 also increases capillary density, again with no significant difference in VEGF levels in the wounds compared

  16. Bacteriophage SP6 encodes a second tailspike protein that recognizes Salmonella enterica serogroups C{sub 2} and C{sub 3}

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

    Gebhart, Dana; Williams, Steven R.; Scholl, Dean,

    SP6 is a salmonella phage closely related to coliphage K1-5. K1-5 is notable in that it encodes two polysaccharide-degrading tailspike proteins, an endosialidase that allows it to infect E. coli K1, and a lyase that enables it to infect K5 strains. SP6 is similar to K1-5 except that it encodes a P22-like endorhamnosidase tailspike, gp46, allowing it to infect group B Salmonella. We show here that SP6 can also infect Salmonella serogroups C{sub 2} and C{sub 3} and that a mutation in a putative second tailspike, gp47, eliminates this specificity. Gene 47 was fused to the coding region of themore » N-terminal portion of the Pseudomonas aeruginosa R2 pyocin tail fiber and expressed in trans such that the fusion protein becomes incorporated into pyocin particles. These pyocins, termed AvR2-SP47, killed serogroups C{sub 2} and C{sub 3}Salmonella. We conclude that SP6 encodes two tail proteins providing it a broad host range among Salmonella enterica. - Highlights: • SP6 is a “dual specificity” bacteriophage that encodes two different receptor binding proteins giving it a broad host range. • These receptor binding proteins can be used to re-target the spectrum of R-type bacteriocins to Salmonella enterica. • Both SP6 and the engineered R-type bacteriocins can kill the Salmonella serovars most associated with human disease making them attractive for development as antimicrobial agents.« less

  17. Adenoviral Mediated Expression of BMP2 by Bone Marrow Stromal Cells Cultured in 3D Copolymer Scaffolds Enhances Bone Formation.

    PubMed

    Sharma, Sunita; Sapkota, Dipak; Xue, Ying; Sun, Yang; Finne-Wistrand, Anna; Bruland, Ove; Mustafa, Kamal

    2016-01-01

    Selection of appropriate osteoinductive growth factors, suitable delivery method and proper supportive scaffold are critical for a successful outcome in bone tissue engineering using bone marrow stromal cells (BMSC). This study examined the molecular and functional effect of a combination of adenoviral mediated expression of bone morphogenetic protein-2 (BMP2) in BMSC and recently developed and characterized, biodegradable Poly(L-lactide-co-є-caprolactone){poly(LLA-co-CL)}scaffolds in osteogenic molecular changes and ectopic bone formation by using in vitro and in vivo approaches. Pathway-focused custom PCR array, validation using TaqMan based quantitative RT-PCR (qRT-PCR) and ALP staining showed significant up-regulation of several osteogenic and angiogenic molecules, including ALPL and RUNX2 in ad-BMP2 BMSC group grown in poly(LLA-co-CL) scaffolds both at 3 and 14 days. Micro CT and histological analyses of the subcutaneously implanted scaffolds in NOD/SCID mice revealed significantly increased radiopaque areas, percentage bone volume and formation of vital bone in ad-BMP2 scaffolds as compared to the control groups both at 2 and 8 weeks. The increased bone formation in the ad-BMP2 group in vivo was paralleled at the molecular level with concomitant over-expression of a number of osteogenic and angiogenic genes including ALPL, RUNX2, SPP1, ANGPT1. The increased bone formation in ad-BMP2 explants was not found to be associated with enhanced endochondral activity as evidenced by qRT-PCR (SOX9 and FGF2) and Safranin O staining. Taken together, combination of adenoviral mediated BMP-2 expression in BMSC grown in the newly developed poly(LLA-co-CL) scaffolds induced expression of osteogenic markers and enhanced bone formation in vivo.

  18. Diagnosing causes of increased NO{sub x} at Potomac River Unit 4

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

    D`Agostini, M.; Levy, E.; Blankenship, D.

    1996-12-31

    Potomac River Unit 4, a 108 MW unit, has a corner-fired boiler with conventional burners. Through the use of combustion optimization techniques, the NO{sub x} emissions had been reduced from baseline levels of 0.6 lb/MBtu to values below 0.45 lb/MBtu over the load range, and had been routinely dispatched at this reduced emissions level for extended periods of operation. After a scheduled outage in Fall 1995 for boiler maintenance, windbox modifications and a superheater replacement, the unit was returned to service, and it was found that the full-load NO{sub x} level had increased by approximately 0.1 lb/MBtu. A week ofmore » tests was performed and pre- and post-outage data analyzed to determine the factors contributing to the increase in NO{sub x} emissions. The principal cause was found to be a reduction in the amount of boiler air in-leakage arising from boiler casing repairs, and leading to higher air-to-fuel ratios in the furnace. Other, secondary, causes were also determined. Results from the analysis indicate the windbox modifications were actually effective in reducing NO{sub x}, although the benefits were obscured due to the intervening circumstances. This paper describes the diagnostic process which was followed and discusses the findings from the investigation.« less

  19. Crystallization and preliminary crystallographic analysis of recombinant immunoglobulin G-binding protein from Streptococcus suis

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

    Khan, Abdul Hamid; Chu, Fuliang; Feng, Youjun

    2008-08-01

    Crystallization of recombinant IgG-binding protein expressed in Escherichia coli using the hanging-drop vapour-diffusion method is described. The crystals belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 38.98, b = 43.94, c = 78.17 Å. Streptococcus suis, an important zoonotic pathogen, expresses immunoglobulin G-binding protein, which is thought to be helpful to the organism in eluding the host defence system. Recombinant IgG-binding protein expressed in Escherichia coli has been crystallized using the hanging-drop vapour-diffusion method. The crystals belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 38.98, b = 43.94, c =more » 78.17 Å and one molecule in the asymmetric unit. Diffraction data were collected to 2.60 Å resolution.« less

  20. Transducing Airway Basal Cells with a Helper-Dependent Adenoviral Vector for Lung Gene Therapy.

    PubMed

    Cao, Huibi; Ouyang, Hong; Grasemann, Hartmut; Bartlett, Claire; Du, Kai; Duan, Rongqi; Shi, Fushan; Estrada, Marvin; Seigel, Kyle E; Coates, Allan L; Yeger, Herman; Bear, Christine E; Gonska, Tanja; Moraes, Theo J; Hu, Jim

    2018-06-01

    A major challenge in developing gene-based therapies for airway diseases such as cystic fibrosis (CF) is sustaining therapeutic levels of transgene expression over time. This is largely due to airway epithelial cell turnover and the host immunogenicity to gene delivery vectors. Modern gene editing tools and delivery vehicles hold great potential for overcoming this challenge. There is currently not much known about how to deliver genes into airway stem cells, of which basal cells are the major type in human airways. In this study, helper-dependent adenoviral (HD-Ad) vectors were delivered to mouse and pig airways via intranasal delivery, and direct bronchoscopic instillation, respectively. Vector transduction was assessed by immunostaining of lung tissue sections, which revealed that airway basal cells of mice and pigs can be targeted in vivo. In addition, efficient transduction of primary human airway basal cells was verified with an HD-Ad vector expressing green fluorescent protein. Furthermore, we successfully delivered the human CFTR gene to airway basal cells from CF patients, and demonstrated restoration of CFTR channel activity following cell differentiation in air-liquid interface culture. Our results provide a strong rationale for utilizing HD-Ad vectors to target airway basal cells for permanent gene correction of genetic airway diseases.

  1. 7 CFR 275.7 - Selection of sub-units for review.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Agriculture Regulations of the Department of Agriculture (Continued) FOOD AND NUTRITION SERVICE, DEPARTMENT OF AGRICULTURE FOOD STAMP AND FOOD DISTRIBUTION PROGRAM PERFORMANCE REPORTING SYSTEM Management Evaluation (ME... Food Stamp Program, exclusive of Post Offices which may issue coupons. Sub-units shall be classified...

  2. Expression of BCL-2 in liver grafts after adenoviral transfer improves survival following prolonged ischemia and reperfusion in rat liver transplantation.

    PubMed

    Kienle, K; Rentsch, M; Müller, T; Engelhard, N; Vogel, M; Jauch, K W; Beham, A

    2005-01-01

    Apoptosis represents a crucial mechanism of ischemia-reperfusion injury after liver transplantation. Bcl-2 may inhibit apoptosis. This study investigates the effect on ischemia/reperfusion injury and survival after rat liver transplantation of adenoviral bcl-2 transfer into donor livers. A nonreplicative adenovirus, expressing bcl-2 under control of a tetracyclin-inducible promoter (adv TetOn bcl-2) was used to treat male Lewis rats in combination with a second adenovirus transferring the TetOn repressor protein under control of a cytomegalovirus promoter (advCMVRep). Virus induction was achieved by addition of doxycyclin to the drinking water. Controls were pretreated with a control adenovirus (advCMV GFP) or with doxycycline. Liver transplantations were performed after 16-hour graft storage. Bcl-2 expression was evaluated by Western blot and immunohistology. Survival was monitored for 7 days, and tissue specimens were collected at 24 hours and 7 days post reperfusion. After pretreatment with advTetOn bcl-2/adv CMVRep, intrahepatic bcl-2 expression was evident at 24 hours and 7 days but was absent among controls. Bcl-2 expression was detected in hepatocytes and, to a high degree, in sinusoidal lining cells. TUNEL-positive sinusoidal lining cells were strikingly reduced after bcl-2 transfer (0.1 +/- 0.3 cells/hpf, mean +/- SD) compared to control virus (4.8 +/- 2.3) or doxycyclin-treated grafts (1.3 +/- 0.2); P < .05. After bcl-2 treatment, survival after transplantation was 100%, whereas it was 50% in both control groups (P = .035). The study shows the feasibility of transient, doxycyclin-controlled adenoviral gene transfer in a transplantation model. Bcl-2 expression increased survival after ischemia/reperfusion in rat liver transplantation, potentially through protection of sinusoidal lining cells.

  3. Nano titanium dioxide photocatalytic protein tyrosine nitration: A potential hazard of TiO{sub 2} on skin

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

    Lu, Naihao; Zhu Zhening; Zhao Xuqi

    Protein tyrosine nitration is a prevalent post-translational modification which occurs as a result of oxidative and nitrative stress, it may be directly involved in the onset and/or progression of diseases. Considering the existence of nano titanium dioxide (TiO{sub 2}) in environment and sunscreen products along with the high content of nitrite in sweat, the UV-exposed skin may be a significant target for the photosensitized damage. In this paper, tyrosine nitration of bovine serum albumin (BSA) was initiated in the UV-irradiated reaction mixture containing 0.2-3.0 mg/ml of three commercially nano TiO{sub 2} products and 0.25-1.0 mM NO{sub 2}{sup -}. It wasmore » found that anatase TiO{sub 2} and Degussa P25 TiO{sub 2} showed prominent photocatalytic activity on promoting the formation of protein tyrosine nitration, and the optimum condition for the reaction was around physiological pH. Meanwhile, the photocatalytic effect of rutile on protein tyrosine nitration was subtle. The potential physiological significance of nano TiO{sub 2}-photocatalytic protein nitration was also demonstrated in mouse skin homogenate. Although the relationship between photocatalytic protein tyrosine nitration and chronic cutaneous diseases needs further study, the toxicity of nano TiO{sub 2} to the skin disease should be paid more attention in the production and utilization process.« less

  4. Chromosomal integration of adenoviral vector DNA in vivo.

    PubMed

    Stephen, Sam Laurel; Montini, Eugenio; Sivanandam, Vijayshankar Ganesh; Al-Dhalimy, Muhseen; Kestler, Hans A; Finegold, Milton; Grompe, Markus; Kochanek, Stefan

    2010-10-01

    So far there has been no report of any clinical or preclinical evidence for chromosomal vector integration following adenovirus (Ad) vector-mediated gene transfer in vivo. We used liver gene transfer with high-capacity Ad vectors in the FAH(Deltaexon5) mouse model to analyze homologous and heterologous recombination events between vector and chromosomal DNA. Intravenous injection of Ad vectors either expressing a fumarylacetoacetate hydrolase (FAH) cDNA or carrying part of the FAH genomic locus resulted in liver nodules of FAH-expressing hepatocytes, demonstrating chromosomal vector integration. Analysis of junctions between vector and chromosomal DNA following heterologous recombination indicated integration of the vector genome through its termini. Heterologous recombination occurred with a median frequency of 6.72 x 10(-5) per transduced hepatocyte, while homologous recombination occurred more rarely with a median frequency of 3.88 x 10(-7). This study has established quantitative and qualitative data on recombination of adenoviral vector DNA with genomic DNA in vivo, contributing to a risk-benefit assessment of the biosafety of Ad vector-mediated gene transfer.

  5. Evaluation of cell-mediated immune responses against porcine circovirus type 2 (PCV2) Cap and Rep proteins after vaccination with a commercial PCV2 sub-unit vaccine.

    PubMed

    Fort, Maria; Sibila, Marina; Nofrarías, Miquel; Pérez-Martín, Eva; Olvera, Alex; Mateu, Enric; Segalés, Joaquim

    2012-11-15

    This study investigated the development of cellular immunity to Porcine circovirus type 2 (PCV2) Cap and Rep proteins in pigs vaccinated with a commercial PCV2 genotype a (PCV2a) based sub-unit vaccine, before and after a heterologous challenge with a PCV2b isolate. At three weeks of age, 20 pigs were inoculated intramuscularly with either the vaccine product (V group, n=9) or phosphate buffered saline solution (PBS) (NV group, n=11). Three weeks after vaccination, pigs were challenged intranasally with PCV2b (V-C and NV-C groups) or PBS (V-NC and NV-NC groups). None of the pigs developed clinical signs during the whole experiment, but all NV-C and 3/5 V-C pigs developed viraemia. Vaccination induced the development IFN-γ-secreting cells in response to the Cap protein of PCV2, which appeared three weeks post-vaccination and increased after challenge. By that time, no significant differences were detected on PCV2 antibody titres between vaccinated and non-vaccinated pigs, although there were significant differences on day 7 post-challenge. PCV2-inoculation induced a cellular response against the Rep protein. Such response was significantly reduced or even absent in PCV2-inoculated pigs that were previously vaccinated (V-C group), presumably as a result of a lower PCV2 replication in vaccinated animals compared to non-vaccinated ones. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. Improved exercise capacity and reduced systemic inflammation after adenoviral-mediated SERCA-2a gene transfer.

    PubMed

    Gupta, Dipin; Palma, Jon; Molina, Ezequiel; Gaughan, John P; Long, Walter; Houser, Steven; Macha, Mahender

    2008-04-01

    We hypothesized that sarcoplasmic reticulum Ca2+ ATPase pump (SERCA-2a) gene delivery would have beneficial effects upon exercise capacity and markers of inflammation in the setting of heart failure. A pressure-overload model of experimental heart failure was used in rats. Following a decrease in fractional shortening of >or=25%, animals underwent intracoronary adenoviral-mediated gene transfection using SERCA-2a. Heart failure animals were randomized to receive the SERCA-2a gene, the beta galactosidase (control) gene, or followed without any further intervention. Exercise and hemodynamic testing were performed, and myocardial and systemic markers of inflammation were assayed after 7 and 21 d. Animals receiving Ad.SERCA-2a showed an increase in exercise tolerance (499.0 +/- 14.9 versus 312.8 +/- 10.5 s, P < 0.0001) relative to Ad.Gal group. Groups treated with Ad.SERCA-2a had significantly decreased serum levels of the inflammatory markers interleukin-1, interleukin-6, and tumor necrosis factor-alpha compared with Ad.Gal-treated animals. Serum levels of atrial natriuretic peptide were decreased in animals receiving Ad.SERCA-2a compared with animals receiving Ad.Gal at day 7 (0.35 +/- 0.03 versus 0.52 +/- 0.11 pg/mL, P = 0.001). Myocardial levels of the proapoptotic protein bax were reduced in Ad.SERCA-2a -treated animals compared with those receiving Ad.Gal at day 7 (protein level/actin: 0.24 +/- 0.05 versus 0.33 +/- 0.04, P = 0.04) and day 21 (protein level/actin: 0.61 +/- 0.04 versus 0.69 +/- 0.01, P = 0.001). Genetic modulation of heart failure using the SERCA-2a gene was associated with improvement in cardiac function and exercise capacity as well as improvements in heart-failure associated inflammatory markers.

  7. STANDARDIZATION AND VALIDATION OF ADENOVIRAL TRANSDUCTION OF AN ANDROGEN RECEPTOR POSITIVE CELL LINE WITH AN MMTV-LUC REPORTER FOR ENDOCRINE SCREENING

    EPA Science Inventory

    Standardization and Validation of Adenoviral Transduction of an Androgen Receptor Positive Cell Line with an MMTV-Luc Reporter for Endocrine Screening P. Hartig, K . Bobseine,
    M. Cardon, C. Lambright and L. E. Gray, Jr. USEPA, Reproductive Toxicology Division, NHEERL, RTP, NC...

  8. PhosphoregDB: The tissue and sub-cellular distribution of mammalian protein kinases and phosphatases

    PubMed Central

    Forrest, Alistair RR; Taylor, Darrin F; Fink, J Lynn; Gongora, M Milena; Flegg, Cameron; Teasdale, Rohan D; Suzuki, Harukazu; Kanamori, Mutsumi; Kai, Chikatoshi; Hayashizaki, Yoshihide; Grimmond, Sean M

    2006-01-01

    Background Protein kinases and protein phosphatases are the fundamental components of phosphorylation dependent protein regulatory systems. We have created a database for the protein kinase-like and phosphatase-like loci of mouse that integrates protein sequence, interaction, classification and pathway information with the results of a systematic screen of their sub-cellular localization and tissue specific expression data mined from the GNF tissue atlas of mouse. Results The database lets users query where a specific kinase or phosphatase is expressed at both the tissue and sub-cellular levels. Similarly the interface allows the user to query by tissue, pathway or sub-cellular localization, to reveal which components are co-expressed or co-localized. A review of their expression reveals 30% of these components are detected in all tissues tested while 70% show some level of tissue restriction. Hierarchical clustering of the expression data reveals that expression of these genes can be used to separate the samples into tissues of related lineage, including 3 larger clusters of nervous tissue, developing embryo and cells of the immune system. By overlaying the expression, sub-cellular localization and classification data we examine correlations between class, specificity and tissue restriction and show that tyrosine kinases are more generally expressed in fewer tissues than serine/threonine kinases. Conclusion Together these data demonstrate that cell type specific systems exist to regulate protein phosphorylation and that for accurate modelling and for determination of enzyme substrate relationships the co-location of components needs to be considered. PMID:16504016

  9. Super-resolution imaging and tracking of protein-protein interactions in sub-diffraction cellular space

    NASA Astrophysics Data System (ADS)

    Liu, Zhen; Xing, Dong; Su, Qian Peter; Zhu, Yun; Zhang, Jiamei; Kong, Xinyu; Xue, Boxin; Wang, Sheng; Sun, Hao; Tao, Yile; Sun, Yujie

    2014-07-01

    Imaging the location and dynamics of individual interacting protein pairs is essential but often difficult because of the fluorescent background from other paired and non-paired molecules, particularly in the sub-diffraction cellular space. Here we develop a new method combining bimolecular fluorescence complementation and photoactivated localization microscopy for super-resolution imaging and single-molecule tracking of specific protein-protein interactions. The method is used to study the interaction of two abundant proteins, MreB and EF-Tu, in Escherichia coli cells. The super-resolution imaging shows interesting distribution and domain sizes of interacting MreB-EF-Tu pairs as a subpopulation of total EF-Tu. The single-molecule tracking of MreB, EF-Tu and MreB-EF-Tu pairs reveals intriguing localization-dependent heterogonous dynamics and provides valuable insights to understanding the roles of MreB-EF-Tu interactions.

  10. Sidestream Smoke Exposure Increases the Susceptibility of Airway Epithelia to Adenoviral Infection

    PubMed Central

    Sharma, Priyanka; Kolawole, Abimbola O.; Core, Susan B.; Kajon, Adriana E.; Excoffon, Katherine J. D. A.

    2012-01-01

    Background Although significant epidemiological evidence indicates that cigarette smoke exposure increases the incidence and severity of viral infection, the molecular mechanisms behind the increased susceptibility of the respiratory tract to viral pathogens are unclear. Adenoviruses are non-enveloped DNA viruses and important causative agents of acute respiratory disease. The Coxsackievirus and adenovirus receptor (CAR) is the primary receptor for many adenoviruses. We hypothesized that cigarette smoke exposure increases epithelial susceptibility to adenovirus infection by increasing the abundance of apical CAR. Methodology and Findings Cultured human airway epithelial cells (CaLu-3) were used as a model to investigate the effect of sidestream cigarette smoke (SSS), mainstream cigarette smoke (MSS), or control air exposure on the susceptibility of polarized respiratory epithelia to adenoviral infection. Using a Cultex air-liquid interface exposure system, we have discovered novel differences in epithelial susceptibility between SSS and MSS exposures. SSS exposure upregulates an eight-exon isoform of CAR and increases adenoviral entry from the apical surface whilst MSS exposure is similar to control air exposure. Additionally, the level of cellular glycogen synthase kinase 3β (GSK3β) is downregulated by SSS exposure and treatment with a specific GSK3β inhibitor recapitulates the effects of SSS exposure on CAR expression and viral infection. Conclusions This is the first time that SSS exposure has been shown to directly enhance the susceptibility of a polarized epithelium to infection by a common respiratory viral pathogen. This work provides a novel understanding of the impact of SSS on the burden of respiratory viral infections and may lead to new strategies to alter viral infections. Moreover, since GSK3β inhibitors are under intense clinical investigation as therapeutics for a diverse range of diseases, studies such as these might provide insight to extend

  11. ERK2-Mediated Phosphorylation of Transcriptional Coactivator Binding Protein PIMT/NCoA6IP at Ser298 Augments Hepatic Gluconeogenesis

    PubMed Central

    Parsa, Kishore V. L.; Kain, Vasundhara; Behera, Soma; Suraj, Sashidhara Kaimal; Babu, Phanithi Prakash; Kar, Anand; Panda, Sunanda; Zhu, Yi-jun; Jia, Yuzhi; Thimmapaya, Bayar; Reddy, Janardan K.; Misra, Parimal

    2013-01-01

    PRIP-Interacting protein with methyl transferase domain (PIMT) serves as a molecular bridge between CREB-binding protein (CBP)/ E1A binding protein p300 (Ep300) -anchored histone acetyl transferase and the Mediator complex sub-unit1 (Med1) and modulates nuclear receptor transcription. Here, we report that ERK2 phosphorylates PIMT at Ser298 and enhances its ability to activate PEPCK promoter. We observed that PIMT is recruited to PEPCK promoter and adenoviral-mediated over-expression of PIMT in rat primary hepatocytes up-regulated expression of gluconeogenic genes including PEPCK. Reporter experiments with phosphomimetic PIMT mutant (PIMTS298D) suggested that conformational change may play an important role in PIMT-dependent PEPCK promoter activity. Overexpression of PIMT and Med1 together augmented hepatic glucose output in an additive manner. Importantly, expression of gluconeogenic genes and hepatic glucose output were suppressed in isolated liver specific PIMT knockout mouse hepatocytes. Furthermore, consistent with reporter experiments, PIMTS298D but not PIMTS298A augmented hepatic glucose output via up-regulating the expression of gluconeogenic genes. Pharmacological blockade of MAPK/ERK pathway using U0126, abolished PIMT/Med1-dependent gluconeogenic program leading to reduced hepatic glucose output. Further, systemic administration of T4 hormone to rats activated ERK1/2 resulting in enhanced PIMT ser298 phosphorylation. Phosphorylation of PIMT led to its increased binding to the PEPCK promoter, increased PEPCK expression and induction of gluconeogenesis in liver. Thus, ERK2-mediated phosphorylation of PIMT at Ser298 is essential in hepatic gluconeogenesis, demonstrating an important role of PIMT in the pathogenesis of hyperglycemia. PMID:24358311

  12. Reverse Monte Carlo simulation of Se{sub 80}Te{sub 20} and Se{sub 80}Te{sub 15}Sb{sub 5} glasses

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

    Abdel-Baset, A. M.; Rashad, M.; Moharram, A. H.

    2013-12-16

    Two-dimensional Monte Carlo of the total pair distribution functions g(r) is determined for Se{sub 80}Te{sub 20} and Se{sub 80}Te{sub 15}Sb{sub 5} alloys, and then it used to assemble the three-dimensional atomic configurations using the reverse Monte Carlo simulation. The partial pair distribution functions g{sub ij}(r) indicate that the basic structure unit in the Se{sub 80}Te{sub 15}Sb{sub 5} glass is di-antimony tri-selenide units connected together through Se-Se and Se-Te chain. The structure of Se{sub 80}Te{sub 20} alloys is a chain of Se-Te and Se-Se in addition to some rings of Se atoms.

  13. Nanoscale organization of {beta}{sub 2}-adrenergic receptor-Venus fusion protein domains on the surface of mammalian cells

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

    Vobornik, Dusan; Rouleau, Yanouchka; Haley, Jennifer

    2009-04-24

    Adrenergic receptors are a key component of nanoscale multiprotein complexes that are responsible for controlling the beat rate in a mammalian heart. We demonstrate the ability of near-field scanning optical microscopy (NSOM) to visualize {beta}{sub 2}-adrenergic receptors ({beta}{sub 2}AR) fused to the GFP analogue Venus at the nanoscale on HEK293 cells. The expression of the {beta}{sub 2}AR-Venus fusion protein was tightly controlled using a tetracycline-induced promoter. Both the size and density of the observed nanoscale domains are dependent on the level of induction and thus the level of protein expression. At concentrations between 100 and 700 ng/ml of inducer doxycycline,more » the size of domains containing the {beta}{sub 2}AR-Venus fusion protein appears to remain roughly constant, but the number of domains per cell increase. At 700 ng/ml doxycycline the functional receptors are organized into domains with an average diameter of 150 nm with a density similar to that observed for the native protein on primary murine cells. By contrast, larger micron-sized domains of {beta}{sub 2}AR are observed in the membrane of the HEK293 cells that stably overexpress {beta}{sub 2}AR-GFP and {beta}{sub 2}AR-eYFP. We conclude that precise chemical control of gene expression is highly advantageous for the use {beta}{sub 2}AR-Venus fusion proteins as models for {beta}{sub 2}AR function. These observations are critical for designing future cell models and assays based on {beta}{sub 2}AR, since the receptor biology is consistent with a relatively low density of nanoscale receptor domains.« less

  14. The S sub 10 /V/ unit of surface brightness. [for zodiacal light measurement

    NASA Technical Reports Server (NTRS)

    Sparrow, J. G.; Weinberg, J. L.

    1976-01-01

    Some discrepancies in the use of the unit of zodiacal light measurements - S sub 10 (V), which is the equivalent number of tenth magnitude stars of solar spectral type per square degree - are discussed. It is suggested that: (1) the S sub 10 (V) unit be understood to represent 10th magnitude solar (G2V) stars per square degree at mean solar distance, (2) the V refers to the visual color in the UBV system defined by Johnson and Morgan (1953), (3) the apparent solar visual magnitude be taken as -26.73 and the B-V index as .63, (4) the solar spectral irradiance values of Labs and Neckel (1970) be used, and (5) when using Vega as a standard to obtain brightnesses in S sub 10 (V), +.04 be used as its magnitude at all wavelengths and the irradiance values of Hayes and Latham (1975) be used.

  15. Synthesis and an X-ray diffraction study of Rb{sub 2}[(UO{sub 2}){sub 2}(C{sub 2}O{sub 4}){sub 3}

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

    Serezhkina, L. B., E-mail: Lserezh@ssu.samara.ru; Peresypkina, E. V.; Neklyudova, N. A.

    2010-09-15

    The synthesis and X-ray diffraction study of compound Rb{sub 2}[(UO{sub 2}){sub 2}(C{sub 2}O{sub 4}){sub 3}], which crystallizes in the monoclinic crystal system, are performed. The unit cell parameters are as follows: a = 7.9996(6) A, b = 8.8259(8) A, c = 11.3220(7) A, {beta} = 105.394(2){sup o}, and V = 770.7(1) A{sup 3}; space group P2{sub 1}/n, Z = 2, and R{sub 1} = 0.0271. [(UO{sub 2}){sub 2}(C{sub 2}O{sub 4}){sub 3}]{sup 2-} layers belonging to the AK{sub 0.5}{sup 02}T{sup 11} crystal chemical group of uranyl complexes (A = UO{sub 2}{sup 2+}, K{sup 02} = C{sub 2}O{sub 4}{sup 2-}, and T{supmore » 11} = C{sub 2}O{sub 4}{sup 2-}) are uranium-containing structural units of the crystals. The layers are connected with outer-sphere rubidium cations by electrostatic interactions.« less

  16. The Nuclear Protein Database (NPD): sub-nuclear localisation and functional annotation of the nuclear proteome

    PubMed Central

    Dellaire, G.; Farrall, R.; Bickmore, W.A.

    2003-01-01

    The Nuclear Protein Database (NPD) is a curated database that contains information on more than 1300 vertebrate proteins that are thought, or are known, to localise to the cell nucleus. Each entry is annotated with information on predicted protein size and isoelectric point, as well as any repeats, motifs or domains within the protein sequence. In addition, information on the sub-nuclear localisation of each protein is provided and the biological and molecular functions are described using Gene Ontology (GO) terms. The database is searchable by keyword, protein name, sub-nuclear compartment and protein domain/motif. Links to other databases are provided (e.g. Entrez, SWISS-PROT, OMIM, PubMed, PubMed Central). Thus, NPD provides a gateway through which the nuclear proteome may be explored. The database can be accessed at http://npd.hgu.mrc.ac.uk and is updated monthly. PMID:12520015

  17. Synthesis, structure and characterization of two new copper(I)-thioarsenates (III) constructed by the [AsS{sub 3}]{sup 3-} and CuS{sub x} units

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

    Yao, Hua-Gang; Ji, Min; Ji, Shou-Hua

    2013-02-15

    Two new copper(I)-thioarsenates(III), CsCu{sub 2}AsS{sub 3}(1) and KCu{sub 4}AsS{sub 4}(2), have been synthesized solvothermally in the presence of copper powder. Compound 1 is built up of trigonal AsS{sub 3} pyramid, tetrahedral CuAsS{sub 3} and CuS{sub 3} moieties forming 4-membered, 5-membered and 6-membered rings. The fusion of these rings produces layered anions [Cu{sub 2}AsS{sub 3}]{sup -}, and cesium cations are located between metal-sulfide layers. The structure of 2 consists of infinite [Cu{sub 2}S{sub 2}]{sup 2-} chain and layered [Cu{sub 6}As{sub 2}S{sub 6}] linked to form a three-dimensional anionic framework, [Cu{sub 4}AsS{sub 4}]{sup -}, and containing channels in which the potassium cationsmore » reside. - Graphical abstract: Qne layered CsCu{sub 2}AsS{sub 3} and one framework KCu{sub 4}AsS{sub 4} possessing noncondensed [AsS{sub 3}]{sup 3-} unit have been synthesized solvothermally. The optical band gaps of the two compounds are 2.3 and 1.8 eV, respectively. Highlights: Black-Right-Pointing-Pointer We obtained two new copper(I)-thioarsenate(III), CsCu{sub 2}AsS{sub 3} and KCu{sub 4}AsS{sub 4}. Black-Right-Pointing-Pointer Both compounds possess noncondensed [AsS{sub 3}]{sup 3-} unit and represent new structure types. Black-Right-Pointing-Pointer The optical band gaps of the two compounds are 2.3 eV and 1.8 eV, respectively.« less

  18. Adenoviral overexpression of Lhx2 attenuates cell viability but does not preserve the stem cell like phenotype of hepatic stellate cells

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

    Genz, Berit; Thomas, Maria; Pützer, Brigitte M.

    2014-11-01

    Hepatic stellate cells (HSC) are well known initiators of hepatic fibrosis. After liver cell damage, HSC transdifferentiate into proliferative myofibroblasts, representing the major source of extracellular matrix in the fibrotic organ. Recent studies also demonstrate a role of HSC as progenitor or stem cell like cells in liver regeneration. Lhx2 is described as stem cell maintaining factor in different organs and as an inhibitory transcription factor in HSC activation. Here we examined whether a continuous expression of Lhx2 in HSC could attenuate their activation and whether Lhx2 could serve as a potential target for antifibrotic gene therapy. Therefore, we evaluatedmore » an adenoviral mediated overexpression of Lhx2 in primary HSC and investigated mRNA expression patterns by qRT-PCR as well as the activation status by different in vitro assays. HSC revealed a marked increase in activation markers like smooth muscle actin alpha (αSMA) and collagen 1α independent from adenoviral transduction. Lhx2 overexpression resulted in attenuated cell viability as shown by a slightly hampered migratory and contractile phenotype of HSC. Expression of stem cell factors or signaling components was also unaffected by Lhx2. Summarizing these results, we found no antifibrotic or stem cell maintaining effect of Lhx2 overexpression in primary HSC. - Highlights: • We performed adenoviral overexpression of Lhx2 in primary hepatic stellate cells. • Hepatic stellate cells expressed stem cell markers during cultivation. • Cell migration and contractility was slightly hampered upon Lhx2 overexpression. • Lhx2 overexpression did not affect stem cell character of hepatic stellate cells.« less

  19. Adenoviral gene transfer of Akt enhances myocardial contractility and intracellular calcium handling

    PubMed Central

    Cittadini, A; Monti, MG; Iaccarino, G; Di Rella, F; Tsichlis, PN; Di Gianni, A; Strömer, H; Sorriento, D; Peschle, C; Trimarco, B; Saccà, L; Condorelli, G

    2010-01-01

    The serine-threonine kinase Akt/PKB mediates stimuli from different classes of cardiomyocyte receptors, including the growth hormone/insulin like growth factor and the β-adrenergic receptors. Whereas the growth-promoting and antiapoptotic properties of Akt activation are well established, little is known about the effects of Akt on myocardial contractility, intracellular calcium (Ca2+) handling, oxygen consumption, and β-adrenergic pathway. To this aim, Sprague–Dawley rats were subjected to a wild-type Akt in vivo adenoviral gene transfer using a catheter-based technique combined with aortopulmonary crossclamping. Left ventricular (LV) contractility and intracellular Ca2+ handling were evaluated in an isolated isovolumic buffer-perfused, aequorin-loaded whole heart preparations 10 days after the surgery. The Ca2+–force relationship was obtained under steady-state conditions in tetanized muscles. No significant hypertrophy was detected in adenovirus with wild-type Akt (Ad.Akt) versus controls rats (LV-to-body weight ratio 2.6±0.2 versus 2.7±0.1 mg/g, controls versus Ad.Akt, P, NS). LV contractility, measured as developed pressure, increased by 41% in Ad.Akt. This was accounted for by both more systolic Ca2+ available to the contractile machinery (+19% versus controls) and by enhanced myofilament Ca2+ responsiveness, documented by an increased maximal Ca2+-activated pressure (+19% versus controls) and a shift to the left of the Ca2+–force relationship. Such increased contractility was paralleled by a slight increase of myocardial oxygen consumption (14%), while titrated dose of dobutamine providing similar inotropic effect augmented oxygen consumption by 39% (P<0.01). Phospholamban, calsequestrin, and ryanodine receptor LV mRNA and protein content were not different among the study groups, while sarcoplasmic reticulum Ca2+ ATPase protein levels were significantly increased in Ad.Akt rats. β-Adrenergic receptor density, affinity, kinase-1 levels, and

  20. Cell lines that support replication of a novel herpes simplex virus 1 U{sub L}31 deletion mutant can properly target U{sub L}34 protein to the nuclear rim in the absence of U{sub L}31

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

    Liang Li; Tanaka, Michiko; Kawaguchi, Yasushi

    2004-11-10

    Previous results indicated that the herpes simplex virus 1 (HSV-1) U{sub L}31 gene is necessary and sufficient for localization of the U{sub L}34 protein exclusively to the nuclear membrane of infected Hep2 cells. In the current studies, a bacterial artificial chromosome containing the entire HSV-1 strain F genome was used to construct a recombinant viral genome in which a gene encoding kanamycin resistance was inserted in place of 262 codons of the 306 codon U{sub L}31 open reading frame. The deletion virus produced virus titers approximately 10- to 50-fold lower in rabbit skin cells, more than 2000-fold lower in Veromore » cells, and more than 1500-fold lower in CV1 cells, compared to a virus bearing a restored U{sub L}31 gene. The replication of the U{sub L}31 deletion virus was restored on U{sub L}31-complementing cell lines derived either from rabbit skin cells or CV1 cells. Confocal microscopy indicated that the majority of U{sub L}34 protein localized aberrantly in the cytoplasm and nucleoplasm of Vero cells and CV1 cells, whereas U{sub L}34 protein localized at the nuclear membrane in rabbit skin cells, and U{sub L}31 complementing CV1 cells infected with the U{sub L}31 deletion virus. We conclude that rabbit skin cells encode a function that allows proper localization of U{sub L}34 protein to the nuclear membrane. We speculate that this function partially complements that of U{sub L}31 and may explain why U{sub L}31 is less critical for replication in rabbit skin cells as opposed to Vero and CV1 cells.« less

  1. Adenoviral vectors elicit humoral immunity against variable loop 2 of clade C HIV-1 gp120 via "Antigen Capsid-Incorporation" strategy.

    PubMed

    Gu, Linlin; Krendelchtchikova, Valentina; Krendelchtchikov, Alexandre; Farrow, Anitra L; Derdeyn, Cynthia A; Matthews, Qiana L

    2016-01-01

    Adenoviral (Ad) vectors in combination with the "Antigen Capsid-Incorporation" strategy have been applied in developing HIV-1 vaccines, due to the vectors׳ abilities in incorporating and inducing immunity of capsid-incorporated antigens. Variable loop 2 (V2)-specific antibodies were suggested in the RV144 trial to correlate with reduced HIV-1 acquisition, which highlights the importance of developing novel HIV-1 vaccines by targeting the V2 loop. Therefore, the V2 loop of HIV-1 has been incorporated into the Ad capsid protein. We generated adenovirus serotype 5 (Ad5) vectors displaying variable loop 2 (V2) of HIV-1 gp120, with the "Antigen Capsid-Incorporation" strategy. To assess the incorporation capabilities on hexon hypervariable region1 (HVR1) and protein IX (pIX), 20aa or full length (43aa) of V2 and V1V2 (67aa) were incorporated, respectively. Immunizations with the recombinant vectors significantly generated antibodies against both linear and discontinuous V2 epitopes. The immunizations generated durable humoral immunity against V2. This study will lead to more stringent development of various serotypes of adenovirus-vectored V2 vaccine candidates, based on breakthroughs regarding the immunogenicity of V2. Copyright © 2015. Published by Elsevier Inc.

  2. The PP4R1 sub-unit of protein phosphatase PP4 is essential for inhibition of NF-κB by merkel polyomavirus small tumour antigen.

    PubMed

    Abdul-Sada, Hussein; Müller, Marietta; Mehta, Rajni; Toth, Rachel; Arthur, J Simon C; Whitehouse, Adrian; Macdonald, Andrew

    2017-04-11

    Merkel cell carcinoma (MCC) is a highly aggressive skin cancer with a high metastatic potential. The majority of MCC cases are caused by the Merkel cell polyomavirus (MCPyV), through expression of the virus-encoded tumour antigens. Whilst mechanisms attributing tumour antigen expression to transformation are being uncovered, little is known of the mechanisms by which MCPyV persists in the host. We previously identified the MCPyV small T antigen (tAg) as a novel inhibitor of nuclear factor kappa B (NF-kB) signalling and a modulator of the host anti-viral response. Here we demonstrate that regulation of NF-kB activation involves a previously undocumented interaction between tAg and regulatory sub-unit 1 of protein phosphatase 4 (PP4R1). Formation of a complex with PP4R1 and PP4c is required to bridge MCPyV tAg to the NEMO adaptor protein, allowing deactivation of the NF-kB pathway. Mutations in MCPyV tAg that fail to interact with components of this complex, or siRNA depletion of PP4R1, prevents tAg-mediated inhibition of NF-kB and pro-inflammatory cytokine production. Comparison of tAg binding partners from other human polyomavirus demonstrates that interactions with NEMO and PP4R1 are unique to MCPyV. Collectively, these data identify PP4R1 as a novel target for virus subversion of the host anti-viral response.

  3. Structural investigation of MO⋅P{sub 2}O{sub 5}⋅Li{sub 2}O (MO = Fe{sub 2}O{sub 3} or V{sub 2}O{sub 5}) glass systems by FTIR spectroscopy

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

    Andronache, Constantin I., E-mail: androtin03@yahoo.com; Racolta, Dania, E-mail: androtin03@yahoo.com

    2014-11-24

    Glasses from the systems xMO⋅(100−x)[P{sub 2}O{sub 5}⋅Li{sub 2}O] (MO = Fe{sub 2}O{sub 3} or V{sub 2}O{sub 5}) with 0 ≤ x ≤ mol % were prepared in the same conditions and characterized by IR spectroscopy. It was established the mode in which both Fe{sub 2}O{sub 3} and V{sub 2}O{sub 5} influences the local structure of these glasses. The iron ions generally modify in a different way the local structure of these glasses then vanadium ions. The results shown that phosphate units are the main structural units of glass system and the iron and vanadium ions are located in the network.

  4. Bone formation in vivo induced by Cbfa1-carrying adenoviral vectors released from a biodegradable porous β-tricalcium phosphate (β-TCP) material.

    PubMed

    Uemura, Toshimasa; Kojima, Hiroko

    2011-06-01

    Overexpression of Cbfa1 (a transcription factor indispensable for osteoblastic differentiation) is expected to induce the formation of bone directly and indirectly in vivo by accelerating osteoblastic differentiation. Adenoviral vectors carrying the cDNA of Cbfa1/til-1(Adv-Cbf1) were allowed to be adsorbed onto porous blocks of β-tricalcium phosphate (β-TCP), a biodegradable ceramic, which were then implanted subcutaneously and orthotopically into bone defects. The adenoviral vectors were released sustainingly by biodegradation, providing long-term expression of the genes. Results of the subcutaneous implantation of Adv-Cbfa1-adsorbed β-TCP/osteoprogenitor cells suggest that a larger amount of bone formed in the pores of the implant than in the control material. Regarding orthotopic implantation into bone defects, the released Adv-Cbfa1 accelerated regeneration in the cortical bone, whereas it induced bone resorption in the marrow cavity. A safer gene transfer using a smaller amount of the vector was achieved using biodegradable porous β-TCP as a carrier.

  5. Bone formation in vivo induced by Cbfa1-carrying adenoviral vectors released from a biodegradable porous β-tricalcium phosphate (β-TCP) material

    NASA Astrophysics Data System (ADS)

    Uemura, Toshimasa; Kojima, Hiroko

    2011-06-01

    Overexpression of Cbfa1 (a transcription factor indispensable for osteoblastic differentiation) is expected to induce the formation of bone directly and indirectly in vivo by accelerating osteoblastic differentiation. Adenoviral vectors carrying the cDNA of Cbfa1/til-1(Adv-Cbf1) were allowed to be adsorbed onto porous blocks of β-tricalcium phosphate (β-TCP), a biodegradable ceramic, which were then implanted subcutaneously and orthotopically into bone defects. The adenoviral vectors were released sustainingly by biodegradation, providing long-term expression of the genes. Results of the subcutaneous implantation of Adv-Cbfa1-adsorbed β-TCP/osteoprogenitor cells suggest that a larger amount of bone formed in the pores of the implant than in the control material. Regarding orthotopic implantation into bone defects, the released Adv-Cbfa1 accelerated regeneration in the cortical bone, whereas it induced bone resorption in the marrow cavity. A safer gene transfer using a smaller amount of the vector was achieved using biodegradable porous β-TCP as a carrier.

  6. Current Advances and Future Challenges in Adenoviral Vector Biology and Targeting

    PubMed Central

    Campos, Samuel K.; Barry, Michael A.

    2008-01-01

    Gene delivery vectors based on Adenoviral (Ad) vectors have enormous potential for the treatment of both hereditary and acquired disease. Detailed structural analysis of the Ad virion, combined with functional studies has broadened our knowledge of the structure/function relationships between Ad vectors and host cells/tissues and substantial achievement has been made towards a thorough understanding of the biology of Ad vectors. The widespread use of Ad vectors for clinical gene therapy is compromised by their inherent immunogenicity. The generation of safer and more effective Ad vectors, targeted to the site of disease, has therefore become a great ambition in the field of Ad vector development. This review provides a synopsis of the structure/function relationships between Ad vectors and host systems and summarizes the many innovative approaches towards achieving Ad vector targeting. PMID:17584037

  7. Peroxisome Proliferator-Activated Receptor Subtype- and Cell-Type-Specific Activation of Genomic Target Genes upon Adenoviral Transgene Delivery

    PubMed Central

    Nielsen, Ronni; Grøntved, Lars; Stunnenberg, Hendrik G.; Mandrup, Susanne

    2006-01-01

    Investigations of the molecular events involved in activation of genomic target genes by peroxisome proliferator-activated receptors (PPARs) have been hampered by the inability to establish a clean on/off state of the receptor in living cells. Here we show that the combination of adenoviral delivery and chromatin immunoprecipitation (ChIP) is ideal for dissecting these mechanisms. Adenoviral delivery of PPARs leads to a rapid and synchronous expression of the PPAR subtypes, establishment of transcriptional active complexes at genomic loci, and immediate activation of even silent target genes. We demonstrate that PPARγ2 possesses considerable ligand-dependent as well as independent transactivation potential and that agonists increase the occupancy of PPARγ2/retinoid X receptor at PPAR response elements. Intriguingly, by direct comparison of the PPARs (α, γ, and β/δ), we show that the subtypes have very different abilities to gain access to target sites and that in general the genomic occupancy correlates with the ability to activate the corresponding target gene. In addition, the specificity and potency of activation by PPAR subtypes are highly dependent on the cell type. Thus, PPAR subtype-specific activation of genomic target genes involves an intricate interplay between the properties of the subtype- and cell-type-specific settings at the individual target loci. PMID:16847324

  8. SubCellProt: predicting protein subcellular localization using machine learning approaches.

    PubMed

    Garg, Prabha; Sharma, Virag; Chaudhari, Pradeep; Roy, Nilanjan

    2009-01-01

    High-throughput genome sequencing projects continue to churn out enormous amounts of raw sequence data. However, most of this raw sequence data is unannotated and, hence, not very useful. Among the various approaches to decipher the function of a protein, one is to determine its localization. Experimental approaches for proteome annotation including determination of a protein's subcellular localizations are very costly and labor intensive. Besides the available experimental methods, in silico methods present alternative approaches to accomplish this task. Here, we present two machine learning approaches for prediction of the subcellular localization of a protein from the primary sequence information. Two machine learning algorithms, k Nearest Neighbor (k-NN) and Probabilistic Neural Network (PNN) were used to classify an unknown protein into one of the 11 subcellular localizations. The final prediction is made on the basis of a consensus of the predictions made by two algorithms and a probability is assigned to it. The results indicate that the primary sequence derived features like amino acid composition, sequence order and physicochemical properties can be used to assign subcellular localization with a fair degree of accuracy. Moreover, with the enhanced accuracy of our approach and the definition of a prediction domain, this method can be used for proteome annotation in a high throughput manner. SubCellProt is available at www.databases.niper.ac.in/SubCellProt.

  9. Yor022c protein is a phospholipase A{sub 1} that localizes to the mitochondrial matrix

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

    Urafuji, Kyosei; Arioka, Manabu

    In mammals, three types of intracellular phospholipase A{sub 1} (iPLA{sub 1}) enzymes have been characterized and are thought to be involved in various cellular processes such as phospholipid metabolism, organelle biogenesis, and membrane trafficking. In this study we analyzed the unique iPLA{sub 1}-like protein, Yor022c, in the budding yeast Saccharomyces cerevisiae. By the mass spectrometry analysis, we demonstrate that Yor022c is actually a phospholipase displaying sn-1-specific activity toward phosphatidylcholine, phosphatidylethanolamine, and phosphatidic acid, generating 2-acyl lysophospholipids. GFP-fused Yor022c co-stained with the mitochondrial dye MitoTracker, indicating that, unlike its mammalian counterparts, it is a mitochondrial protein. Further biochemical fractionation experiment combinedmore » with protease sensitivity assay showed that Yor022c localizes to the mitochondrial matrix. Thus Yor022c is the first PLA{sub 1} putatively involved in the maintenance of sn-1 acyl chains of phospholipids in the mitochondrial inner membrane. - Highlights: • Yeast Yor022c protein displays phospholipase A{sub 1} activity to various phospholipids. • Yor022c-GFP fusion protein localizes to mitochondria. • Biochemical fractionation showed that Yor022c localizes to the mitochondrial matrix.« less

  10. Solvothermal indium fluoride chemistry: Syntheses and crystal structures of K{sub 5}In{sub 3}F{sub 14}, beta-(NH{sub 4}){sub 3}InF{sub 6} and [NH{sub 4}]{sub 3}[C{sub 6}H{sub 21}N{sub 4}]{sub 2}[In{sub 4}F{sub 21}

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

    Jayasundera, Anil C.A.; Goff, Richard J.; Li Yang

    2010-02-15

    The solvothermal syntheses and crystal structures of three indium fluorides are presented. K{sub 5}In{sub 3}F{sub 14} (1) and beta-(NH{sub 4}){sub 3}InF{sub 6} (2) are variants on known inorganic structure types chiolite and cryolite, respectively, with the latter exhibiting a complex and apparently novel structural distortion. [NH{sub 4}]{sub 3}[C{sub 6}H{sub 21}N{sub 4}]{sub 2}[In{sub 4}F{sub 21}] (3) represents a new hybrid composition displaying a unique trimeric metal fluoride building unit. - Graphical abstract: Solvothermal synthesis has been used to prepare three indium fluorides, including a novel hybrid material containing a unique [In{sub 3}F{sub 15}] trimer templated by tren.

  11. Lysyl Oxidase-Like 1 Protein Deficiency Protects Mice from Adenoviral Transforming Growth Factor-β1-induced Pulmonary Fibrosis.

    PubMed

    Bellaye, Pierre-Simon; Shimbori, Chiko; Upagupta, Chandak; Sato, Seidai; Shi, Wei; Gauldie, Jack; Ask, Kjetil; Kolb, Martin

    2018-04-01

    Idiopathic pulmonary fibrosis (IPF) is a progressive disease characterized by excessive deposition of extracellular matrix (ECM) in the lung parenchyma. The abnormal ECM deposition slowly overtakes normal lung tissue, disturbing gas exchange and leading to respiratory failure and death. ECM cross-linking and subsequent stiffening is thought to be a major contributor of disease progression and also promotes the activation of transforming growth factor (TGF)-β1, one of the main profibrotic growth factors. Lysyl oxidase-like (LOXL) 1 belongs to the cross-linking enzyme family and has been shown to be up-regulated in active fibrotic regions of bleomycin-treated mice and patients with IPF. We demonstrate in this study that LOXL1-deficient mice are protected from experimental lung fibrosis induced by overexpression of TGF-β1 using adenoviral (Ad) gene transfer (AdTGF-β1). The lack of LOXL1 prevented accumulation of insoluble cross-linked collagen in the lungs, and therefore limited lung stiffness after AdTGF-β1. In addition, we applied mechanical stretch to lung slices from LOXL1 +/+ and LOXL1 -/- mice treated with AdTGF-β1. Lung stiffness (Young's modulus) of LOXL1 -/- lung slices was significantly lower compared with LOXL1 +/+ lung slices. Moreover, the release of activated TGF-β1 after mechanical stretch was significantly lower in LOXL1 -/- mice compared with LOXL1 +/+ mice after AdTGF-β1. These data support the concept that cross-linking enzyme inhibition represents an interesting therapeutic target for drug development in IPF.

  12. Loss of Endothelial Barrier in Marfan Mice (mgR/mgR) Results in Severe Inflammation after Adenoviral Gene Therapy

    PubMed Central

    Weymann, Alexander; Arif, Rawa; Weber, Antje; Zaradzki, Marcin; Richter, Karsten; Ensminger, Stephan; Robinson, Peter Nicholas; Wagner, Andreas H.; Karck, Matthias; Kallenbach, Klaus

    2016-01-01

    Objectives Marfan syndrome is an autosomal dominant inherited disorder of connective tissue. The vascular complications of Marfan syndrome have the biggest impact on life expectancy. The aorta of Marfan patients reveals degradation of elastin layers caused by increased proteolytic activity of matrix metalloproteinases (MMPs). In this study we performed adenoviral gene transfer of human tissue inhibitor of matrix metalloproteinases-1 (hTIMP-1) in aortic grafts of fibrillin-1 deficient Marfan mice (mgR/mgR) in order to reduce elastolysis. Methods We performed heterotopic infrarenal transplantation of the thoracic aorta in female mice (n = 7 per group). Before implantation, mgR/mgR and wild-type aortas (WT, C57BL/6) were transduced ex vivo with an adenoviral vector coding for human TIMP-1 (Ad.hTIMP-1) or β-galactosidase (Ad.β-Gal). As control mgR/mgR and wild-type aortas received no gene therapy. Thirty days after surgery, overexpression of the transgene was assessed by immunohistochemistry (IHC) and collagen in situ zymography. Histologic staining was performed to investigate inflammation, the neointimal index (NI), and elastin breaks. Endothelial barrier function of native not virus-exposed aortas was evaluated by perfusion of fluorescent albumin and examinations of virus-exposed tissue were performed by transmission electron microscopy (TEM). Results IHC and ISZ revealed sufficient expression of the transgene. Severe cellular inflammation and intima hyperplasia were seen only in adenovirus treated mgR/mgR aortas (Ad.β-Gal, Ad.hTIMP-1 NI: 0.23; 0.43), but not in native and Ad.hTIMP-1 treated WT (NI: 0.01; 0.00). Compared to native mgR/mgR and Ad.hTIMP-1 treated WT aorta, the NI is highly significant greater in Ad.hTIMP-1 transduced mgR/mgR aorta (p = 0.001; p = 0.001). As expected, untreated Marfan grafts showed significant more elastolysis compared to WT (p = 0.001). However, elastolysis in Marfan aortas was not reduced by adenoviral overexpression of hTIMP-1

  13. AMELIORATION OF ETHANOL-INDUCED DYSMORPHOGENESIS BY ADENOVIRAL-MEDIATED CU,ZN-SOD AND MN-SOD EXPRESSION IN NEURULATION STAGED MOUSE EMBRYOS IN VITRO

    EPA Science Inventory

    AMELIORATION OF ETHANOL-INDUCED DYSMORPHOGENESIS BY ADENOVIRAL-MEDIATED Cu,Zn-SOD AND Mn-SOD EXPRESSION IN NEURULATION STAGED MOUSE EMBRYOS IN VITRO. JB Smith1, PC Hartig3, MR Blanton3, KK Sulik1,2, and ES Hunter3. 1Department of Cell and Developmental Biology and 2Bowles Cente...

  14. Role of a guanine nucleotide-binding protein in. cap alpha. /sub 1/-adrenergic receptor-mediated Ca/sup 2 +/ mobilization in DDT/sub 1/ MF-2 cells

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

    Cornett, L.E.; Norris, J.S.

    1987-11-01

    In this study the mechanisms involved in ..cap alpha../sub 1/-adrenergic receptor-mediated Ca/sup 2 +/ mobilization at the level of the plasma membrane were investigated. Stimulation of /sup 45/Ca/sup 2 +/ efflux from saponin-permeabilized DDT/sub 1/ MF-2 cells was observed with the addition of either the ..cap alpha../sub 1/-adrenergic agonist phenylephrine and guanosine-5'-triphosphate or the nonhydrolyzable guanine nucleotide guanylyl-imidodiphosphate. In the presence of (/sup 32/P) NAD, pertussis toxin was found to catalyze ADP-ribosylation of a M/sub r/ = 40,500 (n = 8) peptide in membranes prepared from DDT/sub 1/, MF-2 cells, possibly the ..cap alpha..-subunit of N/sub i/. However, stimulation ofmore » unidirectional /sup 45/Ca/sup 2 +/ efflux by phenylephrine was not affected by previous treatment of cells with 100 ng/ml pertussis toxin. These data suggest that the putative guanine nucleotide-binding protein which couples the ..cap alpha../sub 1/-adrenergic receptor to Ca/sup 2 +/ mobilization in DDT/sub 1/ MF-2 cells is not a pertussis toxin substrate and may possibly be an additional member of guanine nucleotide binding protein family.« less

  15. A three-dimensional coordination polymer based on 1,2,3-triazole-4,5-dicarboxylic acid (H{sub 3}tda): ([Cd{sub 12}(tda){sub 8}(H{sub 2}O){sub 11}] · (H{sub 2}O)6.25){sub n}

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

    Zhou, Xin-Hui, E-mail: iamxhzhou@njupt.edu.cn; Chen, Qiang

    The title coordination polymer ([Cd{sub 12}(tda){sub 8}(H{sub 2}O){sub 11}] · (H{sub 2}O){sub 6.25}){sub n} (H{sub 3}tda = 1,2,3-triazole-4,5-dicarboxylic acid), has been hydrothermally synthesized and structurally characterized by single-crystal X-ray diffraction. Complex crystallizes in orthorhombic sp. gr. Pmn2{sub 1} with Z = 4. The Cd{sub 2} unit doublebridged by one carboxylate oxygen atom and two neighboring nitrogen atoms from the tda{sup 3–} ligands are linked by the tda{sup 3–}ligands to lead to the 2D (4,4) network in the ac plane. The almost coplanar Cd{sub 2}(μ{sub 5}-tda){sub 2} unit comprised of two Cd ions double-bridged by two tda{sup 3–} ligands through themore » neighboring nitrogen atoms is connected with the other four Cd{sub 2}(μ{sub 5}-tda){sub 2} units form the undulating 2D network in the ac plane. The (4,4) networks and undulating 2D networks are alternatively connected along the b axis by the tda{sup 3–} ligands coordinating to the Cd ions to form the 3D framework.« less

  16. Roles for herpes simplex virus type 1 U{sub L}34 and U{sub S}3 proteins in disrupting the nuclear lamina during herpes simplex virus type 1 egress

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

    Bjerke, Susan L.; Roller, Richard J.

    2006-04-10

    Cells infected with wild type HSV-1 showed significant lamin A/C and lamin B rearrangement, while U{sub L}34-null virus-infected cells exhibited few changes in lamin localization, indicating that U{sub L}34 is necessary for lamin disruption. During HSV infection, U{sub S}3 limited the development of disruptions in the lamina, since cells infected with a U{sub S}3-null virus developed large perforations in the lamin layer. U{sub S}3 regulation of lamin disruption does not correlate with the induction of apoptosis. Expression of either U{sub L}34 or U{sub S}3 proteins alone disrupted lamin A/C and lamin B localization. Expression of U{sub L}34 and U{sub S}3more » together had little effect on lamin A/C localization, suggesting a regulatory interaction between the two proteins. The data presented in this paper argue for crucial roles for both U{sub L}34 and U{sub S}3 in regulating the state of the nuclear lamina during viral infection.« less

  17. The trafficking pathway of a wheat storage protein in transgenic rice endosperm.

    PubMed

    Oszvald, Maria; Tamas, Laszlo; Shewry, Peter R; Tosi, Paola

    2014-04-01

    The trafficking of proteins in the endoplasmic reticulum (ER) of plant cells is a topic of considerable interest since this organelle serves as an entry point for proteins destined for other organelles, as well as for the ER itself. In the current work, transgenic rice was used to study the pattern and pathway of deposition of the wheat high molecular weight (HMW) glutenin sub-unit (GS) 1Dx5 within the rice endosperm using specific antibodies to determine whether it is deposited in the same or different protein bodies from the rice storage proteins, and whether it is located in the same or separate phases within these. The protein distribution and the expression pattern of HMW sub-unit 1Dx5 in transgenic rice endosperm at different stages of development were determined using light and electron microscopy after labelling with antibodies. The use of HMW-GS-specific antibodies showed that sub-unit 1Dx5 was expressed mainly in the sub-aleurone cells of the endosperm and that it was deposited in both types of protein body present in the rice endosperm: derived from the ER and containing prolamins, and derived from the vacuole and containing glutelins. In addition, new types of protein bodies were also formed within the endosperm cells. The results suggest that the HMW 1Dx5 protein could be trafficked by either the ER or vacuolar pathway, possibly depending on the stage of development, and that its accumulation in the rice endosperm could compromise the structural integrity of protein bodies and their segregation into two distinct populations in the mature endosperm.

  18. Probing GATA factor function in mouse Leydig cells via testicular injection of adenoviral vectors.

    PubMed

    Penny, Gervette M; Cochran, Rebecca B; Pihlajoki, Marjut; Kyrönlahti, Antti; Schrade, Anja; Häkkinen, Merja; Toppari, Jorma; Heikinheimo, Markku; Wilson, David B

    2017-10-01

    Testicular Leydig cells produce androgens essential for proper male reproductive development and fertility. Here, we describe a new Leydig cell ablation model based on Cre/Lox recombination of mouse Gata4 and Gata6 , two genes implicated in the transcriptional regulation of steroidogenesis. The testicular interstitium of adult Gata4 flox/flox ; Gata6 flox/flox mice was injected with adenoviral vectors encoding Cre + GFP (Ad-Cre-IRES-GFP) or GFP alone (Ad-GFP). The vectors efficiently and selectively transduced Leydig cells, as evidenced by GFP reporter expression. Three days after Ad-Cre-IRES-GFP injection, expression of androgen biosynthetic genes ( Hsd3b1 , Cyp17a1 and Hsd17b3 ) was reduced, whereas expression of another Leydig cell marker, Insl3 , was unchanged. Six days after Ad-Cre-IRES-GFP treatment, the testicular interstitium was devoid of Leydig cells, and there was a concomitant loss of all Leydig cell markers. Chromatin condensation, nuclear fragmentation, mitochondrial swelling, and other ultrastructural changes were evident in the degenerating Leydig cells. Liquid chromatography-tandem mass spectrometry demonstrated reduced levels of androstenedione and testosterone in testes from mice injected with Ad-Cre-IRES-GFP. Late effects of treatment included testicular atrophy, infertility and the accumulation of lymphoid cells in the testicular interstitium. We conclude that adenoviral-mediated gene delivery is an expeditious way to probe Leydig cell function in vivo Our findings reinforce the notion that GATA factors are key regulators of steroidogenesis and testicular somatic cell survival.Free Finnish abstract: A Finnish translation of this abstract is freely available at http://www.reproduction-online.org/content/154/4/455/suppl/DC2. © 2017 Society for Reproduction and Fertility.

  19. Mapping proteins in the presence of paralogs using units of coevolution

    PubMed Central

    2013-01-01

    Background We study the problem of mapping proteins between two protein families in the presence of paralogs. This problem occurs as a difficult subproblem in coevolution-based computational approaches for protein-protein interaction prediction. Results Similar to prior approaches, our method is based on the idea that coevolution implies equal rates of sequence evolution among the interacting proteins, and we provide a first attempt to quantify this notion in a formal statistical manner. We call the units that are central to this quantification scheme the units of coevolution. A unit consists of two mapped protein pairs and its score quantifies the coevolution of the pairs. This quantification allows us to provide a maximum likelihood formulation of the paralog mapping problem and to cast it into a binary quadratic programming formulation. Conclusion CUPID, our software tool based on a Lagrangian relaxation of this formulation, makes it, for the first time, possible to compute state-of-the-art quality pairings in a few minutes of runtime. In summary, we suggest a novel alternative to the earlier available approaches, which is statistically sound and computationally feasible. PMID:24564758

  20. Suppression of adenoviral gene expression in the liver: role of innate vs adaptive immunity and their cell lysis mechanisms.

    PubMed

    Minagawa, Masahiro; Kawamura, Hiroki; Liu, Zhangxu; Govindarajan, Sugantha; Dennert, Gunther

    2005-06-01

    Injection of adenoviral constructs causes liver infection prompting immunity, which suppress viral gene expression. Innate and adaptive immunity mediate these processes raising the question which pathways are the most prominent. Adenovirus expressing the beta-galactosidase (beta-gal) gene was injected into normal and immunodeficient mice. Elimination of beta-gal-expressing hepatocytes and increases in liver enzymes were assayed. Major histocompatibility complex (MHC) class I densities, perforin channel insertion and apoptosis by Fas and tumor necrosis factor (TNF)-alpha were assayed. At high virus doses, suppression of viral gene expression was as efficient in immunodeficient as in normal mice, while at low doses effects of cytotoxic T lymphocytes (CTL) were demonstrable. Despite CTL priming and elimination of infected hepatocytes no liver injury is detected. Hepatocyte MHC I densities were able to trigger CTL granule exocytosis and perforin lysis in vitro but not in vivo. This is we show is because of decreased sensitivity of hepatocytes from infected mice to perforin and increased sensitivity to Fas and TNF-alpha lysis. Effector cells of the innate immune system are exceedingly effective in suppressing adenoviral gene expression. Perforin-independent pathways, those mediated by TNF-alpha and Fas are very efficient in hepatocytes from virus-infected livers.

  1. Adenoviral hemorrhagic disease in California mule deer, 1990-2014.

    PubMed

    Woods, Leslie W; Schumaker, Brant A; Pesavento, Patricia A; Crossley, Beate M; Swift, Pamela K

    2018-03-01

    We reviewed case records from the California Animal Health and Food Safety (CAHFS) laboratory and the California Department of Fish and Wildlife (CDFW) spanning 25 years (1990-2014) for all deer accessions submitted to CAHFS for pathology and/or histopathology, with and without a diagnosis of adenoviral hemorrhagic disease (AHD), in order to determine the prevalence of AHD in California. We also examined spatial and temporal distribution, age, and mule deer subspecies in deer that died from AHD. Of 483 deer submitted to CAHFS for diagnostic testing in 1990-2014, 17.2% were diagnosed with confirmed AHD, and 26.5% were confirmed plus suspected cases of AHD. Columbian black-tailed deer ( Odocoileus hemionus columbianus), particularly fawns and juveniles, were most frequently affected. Deer adenovirus ( Odocoileus adenovirus 1; OdAdV-1) was detected by immunohistochemistry in archived CDFW formalin-fixed, paraffin-embedded tissues from deer that died in mortality events in 1981, 1983, and 1986-1987. OdAdV-1 is a common cause of hemorrhagic disease mortality events in California deer, and mortality as a result of AHD is documented as early as 1981.

  2. M/sub r/ 25,000 heparin-binding protein from guinea pig brain is a high molecular weight form of basic fibroblast growth factor

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

    Moscatelli, D.; Joseph-Silverstein, J.; Manejias, R.

    1987-08-01

    A M/sub r/ 25,000 form of basic fibroblast growth factor (bFGF) has been isolated from guinea pig grain along with the typical M/sub r/ 18,000 form. Both forms were purified to homogeneity by a combination of heparin-affinity chromatography and ion-exchange chromatography on an FPLC Mono S column. The M/sub r/ 25,000 form, like the M/sub r/ 18,000 form was not eluted from the heparin-affinity column with 0.95 M NaCl, but was eluted with 2 M NaCl. The M/sub r/ 25,000 guinea pig protein stimulated plasminogen activator production by cultured bovine capillary endothelial cells in a dose-dependent manner at concentration ofmore » 0.1-10 ngml, the same range that was effective for guinea pig and human M/sub r/ 18,000 bFGFs. The binding of human /sup 125/I-labeled bFGF to baby hamster kidney cells is inhibited equally by the M/sub r/ 25,000 guinea pig protein and the M/sub r/ 18,000 guinea pig and human bFGFs. Polyclonal antibodies raised against human bFGF recognize both the M/sub r/ 25,000 and 18,000 guinea pig proteins in an immunoblot analysis. In a radioimmunoassay, both the M/sub r/ 25,000 and M/sub r/ 18,000 guinea pig proteins compete equally well with iodinated human bFGF for binding to the anti-human bFGF antibodies. When treated with low concentrations of trypsin, the M/sub r/ 25,000 guinea pig bFGF was converted to a M/sub r/ 18,000 protein. These results show that the two molecules are closely related and suggest that the M/sub r/ 25,000 protein shares substantial homology with the M/sub r/ 18,000 bFGF« less

  3. Adenovirus E1B 19-Kilodalton Protein Modulates Innate Immunity through Apoptotic Mimicry

    PubMed Central

    Grigera, Fernando; Ucker, David S.; Cook, James L.

    2014-01-01

    ABSTRACT Cells that undergo apoptosis in response to chemical or physical stimuli repress inflammatory reactions, but cells that undergo nonapoptotic death in response to such stimuli lack this activity. Whether cells dying from viral infection exhibit a cell death-type modulatory effect on inflammatory reactions is unknown. We compared the effects on macrophage inflammatory responses of cells dying an apoptotic or a nonapoptotic death as a result of adenoviral infection. The results were exactly opposite to the predictions from the conventional paradigm. Cells dying by apoptosis induced by infection with an adenovirus type 5 (Ad5) E1B 19-kilodalton (E1B 19K) gene deletion mutant did not repress macrophage NF-κB activation or cytokine responses to proinflammatory stimuli, whereas cells dying a nonapoptotic death from infection with E1B 19K-competent, wild-type Ad5 repressed these macrophage inflammatory responses as well as cells undergoing classical apoptosis in response to chemical injury. The immunorepressive, E1B 19K-related cell death activity depended upon direct contact of the virally infected corpses with responder macrophages. Replacement of the viral E1B 19K gene with the mammalian Bcl-2 gene in cis restored the nonapoptotic, immunorepressive cell death activity of virally infected cells. These results define a novel function of the antiapoptotic, adenoviral E1B 19K protein that may limit local host innate immune inflammation during accumulation of virally infected cells at sites of infection and suggest that E1B 19K-deleted, replicating adenoviral vectors might induce greater inflammatory responses to virally infected cells than E1B 19K-positive vectors, because of the net effect of their loss-of-function mutation. IMPORTANCE We observed that cells dying a nonapoptotic cell death induced by adenovirus infection repressed macrophage proinflammatory responses while cells dying by apoptosis induced by infection with an E1B 19K deletion mutant virus did not

  4. Fluorescent protein vectors for pancreatic islet cell identification in live-cell imaging.

    PubMed

    Shuai, Hongyan; Xu, Yunjian; Yu, Qian; Gylfe, Erik; Tengholm, Anders

    2016-10-01

    The islets of Langerhans contain different types of endocrine cells, which are crucial for glucose homeostasis. β- and α-cells that release insulin and glucagon, respectively, are most abundant, whereas somatostatin-producing δ-cells and particularly pancreatic polypeptide-releasing PP-cells are more scarce. Studies of islet cell function are hampered by difficulties to identify the different cell types, especially in live-cell imaging experiments when immunostaining is unsuitable. The aim of the present study was to create a set of vectors for fluorescent protein expression with cell-type-specific promoters and evaluate their applicability in functional islet imaging. We constructed six adenoviral vectors for expression of red and green fluorescent proteins controlled by the insulin, preproglucagon, somatostatin, or pancreatic polypeptide promoters. After transduction of mouse and human islets or dispersed islet cells, a majority of the fluorescent cells also immunostained for the appropriate hormone. Recordings of the sub-plasma membrane Ca(2+) and cAMP concentrations with a fluorescent indicator and a protein biosensor, respectively, showed that labeled cells respond to glucose and other modulators of secretion and revealed a striking variability in Ca(2+) signaling among α-cells. The measurements allowed comparison of the phase relationship of Ca(2+) oscillations between different types of cells within intact islets. We conclude that the fluorescent protein vectors allow easy identification of specific islet cell types and can be used in live-cell imaging together with organic dyes and genetically encoded biosensors. This approach will facilitate studies of normal islet physiology and help to clarify molecular defects and disturbed cell interactions in diabetic islets.

  5. Protein Sub-Nuclear Localization Based on Effective Fusion Representations and Dimension Reduction Algorithm LDA.

    PubMed

    Wang, Shunfang; Liu, Shuhui

    2015-12-19

    An effective representation of a protein sequence plays a crucial role in protein sub-nuclear localization. The existing representations, such as dipeptide composition (DipC), pseudo-amino acid composition (PseAAC) and position specific scoring matrix (PSSM), are insufficient to represent protein sequence due to their single perspectives. Thus, this paper proposes two fusion feature representations of DipPSSM and PseAAPSSM to integrate PSSM with DipC and PseAAC, respectively. When constructing each fusion representation, we introduce the balance factors to value the importance of its components. The optimal values of the balance factors are sought by genetic algorithm. Due to the high dimensionality of the proposed representations, linear discriminant analysis (LDA) is used to find its important low dimensional structure, which is essential for classification and location prediction. The numerical experiments on two public datasets with KNN classifier and cross-validation tests showed that in terms of the common indexes of sensitivity, specificity, accuracy and MCC, the proposed fusing representations outperform the traditional representations in protein sub-nuclear localization, and the representation treated by LDA outperforms the untreated one.

  6. Expression, purification and crystallization of a human protein SH3BGRL at atomic resolution

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

    Yin, Lei; Zhu, De-Yu; Yang, Na

    2005-04-01

    The protein SH3BGRL, containing both SH3-binding and Homer EVH1-binding motifs, has been crystallized using the hanging-drop vapour-diffusion method. The protein SH3BGRL, containing both SH3-binding and Homer EVH1-binding motifs, has been crystallized using the hanging-drop vapour-diffusion method. The crystals diffract to 0.88 Å resolution and belong to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 28.8886, b = 34.9676, c = 98.0016 Å. Preliminary analysis indicates that the asymmetric unit contains one molecule and has a solvent content of about 34%.

  7. INGN 201: Ad-p53, Ad5CMV-p53, adenoviral p53, p53 gene therapy--introgen, RPR/INGN 201.

    PubMed

    2007-01-01

    Introgen and its wholly owned European subsidiary Gendux AB are developing an adenoviral p53 gene therapy as a treatment for cancer in the US and Europe, respectively. Phase III trials in patients with head and neck cancer are ongoing, and a number of clinical trials in other cancer indications have been completed. INGN 201 is being reviewed by the EMEA for approval in Li-Fraumeni syndrome (LFS) under the provisions of exceptional circumstance; the therapy is available on a compassionate use basis to eligible LFS cancer patients under a protocol authorised by the US FDA. The p53 tumour suppressor gene is deleted or mutated in many tumour cells and is one of the most frequently mutated genes in human tumours. The p53 protein is one of the most intricate elements in the apoptotic signalling cascade, and a mutation in the gene encoding it is believed to result in a decreased ability of a cell to apoptose. Thus replacing this gene via adenovirally-mediated p53 gene therapy is hoped to result in increased apoptosis where it is administered.INGN 201 is available for licensing, although Introgen favours retaining partial or full rights to the therapy in the US. Introgen entered into a license agreement with The University of Texas System and MD Anderson Cancer Center in 1994. The technologies licenced include p53 and fus1 (INGN 401). The collaboration has yielded exclusive patent and licensing rights to numerous technologies. Introgen entered into a collaboration with Rhône-Poulenc Rorer Pharmaceuticals (now sanofi-aventis) to develop therapeutics based on p53 inhibition in October 1994. However, in June 2001 this relationship was restructured and Introgen assumed responsibility for the worldwide development of all p53 products including INGN 201, and acquired all marketing and commercialisation rights with respect to those products. Introgen initiated two phase III trials in head and neck cancer (in June 2000 and May 2001) at about 80 sites in the US, Canada and Europe

  8. Role of NH{sub 2}-terminal hydrophobic motif in the subcellular localization of ATP-binding cassette protein subfamily D: Common features in eukaryotic organisms

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

    Lee, Asaka; Asahina, Kota; Okamoto, Takumi

    Highlights: • ABCD proteins classifies based on with or without NH{sub 2}-terminal hydrophobic segment. • The ABCD proteins with the segment are targeted peroxisomes. • The ABCD proteins without the segment are targeted to the endoplasmic reticulum. • The role of the segment in organelle targeting is conserved in eukaryotic organisms. - Abstract: In mammals, four ATP-binding cassette (ABC) proteins belonging to subfamily D have been identified. ABCD1–3 possesses the NH{sub 2}-terminal hydrophobic region and are targeted to peroxisomes, while ABCD4 lacking the region is targeted to the endoplasmic reticulum (ER). Based on hydropathy plot analysis, we found that severalmore » eukaryotes have ABCD protein homologs lacking the NH{sub 2}-terminal hydrophobic segment (H0 motif). To investigate whether the role of the NH{sub 2}-terminal H0 motif in subcellular localization is conserved across species, we expressed ABCD proteins from several species (metazoan, plant and fungi) in fusion with GFP in CHO cells and examined their subcellular localization. ABCD proteins possessing the NH{sub 2}-terminal H0 motif were localized to peroxisomes, while ABCD proteins lacking this region lost this capacity. In addition, the deletion of the NH{sub 2}-terminal H0 motif of ABCD protein resulted in their localization to the ER. These results suggest that the role of the NH{sub 2}-terminal H0 motif in organelle targeting is widely conserved in living organisms.« less

  9. Structure determination of {alpha}-La{sub 6}W{sub 2}O{sub 15}

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

    Chambrier, M-H., E-mail: marie-helene.chambier.etu@univ-lemans.f; Ibberson, R.M.; Goutenoire, F., E-mail: francois.goutenoire@univ-lemans.f

    2010-06-15

    The structure of the high temperature alpha form of La{sub 6}W{sub 2}O{sub 15} has been determined ab-initio from high temperature laboratory X-ray and neutron time-of-flight data. This tungstate crystallizes in the non-centrosymmetric orthorhombic space group (No. 20) C222{sub 1}, with Z=2, a=12.6250(2) A, b=9.1875(1) A, c=5.9688(1) A. The structure comprises [O{sub 2}La{sub 3}] infinite ribbons and is better described by the structural formula [O{sub 2}La{sub 3}]{sub 2}[WO{sub 5.5}]{sub 2}. Using this description we can understand the strong structural similarity of the present compound with compounds of the general composition BiM{sub 2}AO{sub 6} (M=Cu, Mg, Zn, Mn, Cd, Ca, Pb; A=P,more » As, V) described as [O{sub 2}M{sub 2}Bi][AO{sub 4}]. The [WO{sub 5.5}] entity implies oxygen disorder in the material. - Graphical abstract: Projection of the polyhedra around the tungsten atom. Atoms labelled in grey are occupied at 50%. Short oxygen-oxygen distances are marked. The polyhedra represents WO{sub 5.5} that is related to the structural unit W{sub 2}O{sub 11} unit. Alpha-La{sub 6}W{sub 2}O{sub 15} could be described as [O{sub 2}La{sub 3}]{sub 2}[WO{sub 5.5}]{sub 2}.« less

  10. Protein Sub-Nuclear Localization Based on Effective Fusion Representations and Dimension Reduction Algorithm LDA

    PubMed Central

    Wang, Shunfang; Liu, Shuhui

    2015-01-01

    An effective representation of a protein sequence plays a crucial role in protein sub-nuclear localization. The existing representations, such as dipeptide composition (DipC), pseudo-amino acid composition (PseAAC) and position specific scoring matrix (PSSM), are insufficient to represent protein sequence due to their single perspectives. Thus, this paper proposes two fusion feature representations of DipPSSM and PseAAPSSM to integrate PSSM with DipC and PseAAC, respectively. When constructing each fusion representation, we introduce the balance factors to value the importance of its components. The optimal values of the balance factors are sought by genetic algorithm. Due to the high dimensionality of the proposed representations, linear discriminant analysis (LDA) is used to find its important low dimensional structure, which is essential for classification and location prediction. The numerical experiments on two public datasets with KNN classifier and cross-validation tests showed that in terms of the common indexes of sensitivity, specificity, accuracy and MCC, the proposed fusing representations outperform the traditional representations in protein sub-nuclear localization, and the representation treated by LDA outperforms the untreated one. PMID:26703574

  11. RepeatsDB-lite: a web server for unit annotation of tandem repeat proteins.

    PubMed

    Hirsh, Layla; Paladin, Lisanna; Piovesan, Damiano; Tosatto, Silvio C E

    2018-05-09

    RepeatsDB-lite (http://protein.bio.unipd.it/repeatsdb-lite) is a web server for the prediction of repetitive structural elements and units in tandem repeat (TR) proteins. TRs are a widespread but poorly annotated class of non-globular proteins carrying heterogeneous functions. RepeatsDB-lite extends the prediction to all TR types and strongly improves the performance both in terms of computational time and accuracy over previous methods, with precision above 95% for solenoid structures. The algorithm exploits an improved TR unit library derived from the RepeatsDB database to perform an iterative structural search and assignment. The web interface provides tools for analyzing the evolutionary relationships between units and manually refine the prediction by changing unit positions and protein classification. An all-against-all structure-based sequence similarity matrix is calculated and visualized in real-time for every user edit. Reviewed predictions can be submitted to RepeatsDB for review and inclusion.

  12. Anti-Inflammatory Effects of Modified Adenoviral Vectors for Gene Therapy: A View through Animal Models Tested.

    PubMed

    Castañeda-Lopez, M E; Garza-Veloz, I; Lopez-Hernandez, Y; Barbosa-Cisneros, O Y; Martinez-Fierro, M L

    2016-07-01

    The central dogma of gene therapy relies on the application of novel therapeutic genes to treat or prevent diseases. The main types of vectors used for gene transfer are adenovirus, retrovirus, lentivirus, liposome, and adeno-associated virus vectors. Gene therapy has emerged as a promising alternative for the treatment of inflammatory diseases. The main targets are cytokines, co-stimulatory molecules, and different types of cells from hematological and mesenchymal sources. In this review, we focus on molecules with anti-inflammatory effects used for in vivo gene therapy mediated by adenoviral gene transfer in the treatment of immune-mediated inflammatory diseases, with particular emphasis on autoinflammatory and autoimmune diseases.

  13. High temperature redox reactions with uranium: Synthesis and characterization of Cs(UO{sub 2})Cl(SeO{sub 3}), Rb{sub 2}(UO{sub 2}){sub 3}O{sub 2}(SeO{sub 3}){sub 2}, and RbNa{sub 5}U{sub 2}(SO{sub 4}){sub 7}

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

    Babo, Jean-Marie; Department of Civil and Environmental Engineering and Earth Sciences and Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556; Albrecht-Schmitt, Thomas E., E-mail: talbrechtschmitt@gmail.com

    2013-10-15

    Cs(UO{sub 2})Cl(SeO{sub 3}) (1), Rb{sub 2}(UO{sub 2}){sub 3}O{sub 2}(SeO{sub 3}){sub 3} (2), and RbNa{sub 5}U{sub 2}(SO{sub 4}){sub 7} (3) single crystals were synthesized using CsCl, RbCl, and a CuCl/NaCl eutectic mixture as fluxes, respectively. Their lattice parameters and space groups are as follows: P2{sub 1}/n (a=6.548(1) Å, b=11.052(2) Å, c=10.666(2) Å and β=93.897(3)°), P1{sup ¯} (a=7.051(2) Å, b=7.198(2) Å, c=8.314(2) Å, α=107.897(3)°, β=102.687(3)° and γ=100.564(3)°) and C2/c (a=17.862(4) Å, b=6.931(1) Å, c=20.133(4) Å and β=109.737(6)°. The small anionic building units found in these compounds are SeO{sub 3}{sup 2−} and SO{sub 4}{sup 2−} tetrahedra, oxide, and chloride. The crystal structure ofmore » the first compound is composed of [(UO{sub 2}){sub 2}Cl{sub 2}(SeO{sub 3}){sub 2}]{sup 2−} chains separated by Cs{sup +} cations. The structure of (2) is constructed from [(UO{sub 2}){sub 3}O{sub 11}]{sup 16−} chains further connected through selenite units into layers stacked perpendicularly to the [0 1 0] direction, with Rb{sup +} cations intercalating between them. The structure of compound (3) is made of uranyl sulfate layers formed by edge and vertex connections between dimeric [U{sub 2}O{sub 16}] and [SO{sub 4}] polyhedra. These layers contain unusual sulfate–metal connectivity as well as large voids. - Graphical abstract: A new family of uranyl selenites and sulfates has been prepared by high-temperature redox reactions. This compounds display new bonding motifs. Display Omitted - Highlights: • Low-dimensional Uranyl Oxoanion compounds. • Conversion of U(IV) to U(VI) at high temperatures. • Dimensional reduction by both halides and stereochemically active lone-pairs.« less

  14. Structure and magnetic properties of Sm{sub 3}(Fe{sub 0.85}Cr{sub 0.15}){sub 29}N{sub y} alloys

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

    Suzuki, Shunji; Suzuki, Shinya; Kawasaki, Masahito

    1995-11-01

    The structure and magnetic properties of Sm{sub 3}(Fe{sub 0.85}Cr{sub 0.15}){sub 29}N{sub y} alloy were investigated. It was found that the alloy had the R{sub 3}(Fe,M){sub 29}-type structure and absorbed 4--5 wt.% of nitrogen which exceeds 4 nitrogen atoms per unit formula. The coercivity of the powder was greatly influenced by the nitrogen content and annealing. The Sm{sub 3}(Fe{sub 0.85}Cr{sub 0.15}){sub 29}N{sub y} alloy powder with 4.1 wt.% of nitrogen showed a high coercivity of 552 kA/m without fine grinding.

  15. Malonyl-CoA decarboxylase (MCD) is differentially regulated in subcellular compartments by 5'AMP-activated protein kinase (AMPK). Studies using H9c2 cells overexpressing MCD and AMPK by adenoviral gene transfer technique.

    PubMed

    Sambandam, Nandakumar; Steinmetz, Michael; Chu, Angel; Altarejos, Judith Y; Dyck, Jason R B; Lopaschuk, Gary D

    2004-07-01

    Malonyl-CoA, a potent inhibitor of carnitine pamitoyl transferase-I (CPT-I), plays a pivotal role in fuel selection in cardiac muscle. Malonyl-CoA decarboxylase (MCD) catalyzes the degradation of malonyl-CoA, removes a potent allosteric inhibition on CPT-I and thereby increases fatty acid oxidation in the heart. Although MCD has several Ser/Thr phosphorylation sites, whether it is regulated by AMP-activated protein kinase (AMPK) has been controversial. We therefore overexpressed MCD (Ad.MCD) and constitutively active AMPK (Ad.CA-AMPK) in H9c2 cells, using an adenoviral gene delivery approach in order to examine if MCD is regulated by AMPK. Cells infected with Ad.CA-AMPK demonstrated a fourfold increase in AMPK activity as compared with control cells expressing green fluorescent protein (Ad.GFP). MCD activity increased 40- to 50-fold in Ad.MCD + Ad.GFP cells when compared with Ad.GFP control. Co-expressing AMPK with MCD further augmented MCD expression and activity in Ad.MCD + Ad.CA-AMPK cells compared with the Ad.MCD + Ad.GFP control. Subcellular fractionation further revealed that 54.7 kDa isoform of MCD expression was significantly higher in cytosolic fractions of Ad.MCD + Ad.CA-AMPK cells than of the Ad.MCD +Ad.GFP control. However, the MCD activities in cytosolic fractions were not different between the two groups. Interestingly, in the mitochondrial fractions, MCD activity significantly increased in Ad.MCD + Ad.CA-AMPK cells when compared with Ad.MCD + Ad.GFP cells. Using phosphoserine and phosphothreonine antibodies, no phosphorylation of MCD by AMPK was observed. The increase in MCD activity in mitochondria-rich fractions of Ad.MCD + Ad.CA-AMPK cells was accompanied by an increase in the level of the 50.7 kDa isoform of MCD protein in the mitochondria. This differential regulation of MCD expression and activity in the mitochondria by AMPK may potentially regulate malonyl-CoA levels at sites nearby CPT-I on the mitochondria.

  16. A Plasmodium Promiscuous T Cell Epitope Delivered within the Ad5 Hexon Protein Enhances the Protective Efficacy of a Protein Based Malaria Vaccine.

    PubMed

    Fonseca, Jairo Andres; Cabrera-Mora, Monica; Kashentseva, Elena A; Villegas, John Paul; Fernandez, Alejandra; Van Pelt, Amelia; Dmitriev, Igor P; Curiel, David T; Moreno, Alberto

    2016-01-01

    A malaria vaccine is a public health priority. In order to produce an effective vaccine, a multistage approach targeting both the blood and the liver stage infection is desirable. The vaccine candidates also need to induce balanced immune responses including antibodies, CD4+ and CD8+ T cells. Protein-based subunit vaccines like RTS,S are able to induce strong antibody response but poor cellular reactivity. Adenoviral vectors have been effective inducing protective CD8+ T cell responses in several models including malaria; nonetheless this vaccine platform exhibits a limited induction of humoral immune responses. Two approaches have been used to improve the humoral immunogenicity of recombinant adenovirus vectors, the use of heterologous prime-boost regimens with recombinant proteins or the genetic modification of the hypervariable regions (HVR) of the capsid protein hexon to express B cell epitopes of interest. In this study, we describe the development of capsid modified Ad5 vectors that express a promiscuous Plasmodium yoelii T helper epitope denominated PyT53 within the hexon HVR2 region. Several regimens were tested in mice to determine the relevance of the hexon modification in enhancing protective immune responses induced by the previously described protein-based multi-stage experimental vaccine PyCMP. A heterologous prime-boost immunization regime that combines a hexon modified vector with transgenic expression of PyCMP followed by protein immunizations resulted in the induction of robust antibody and cellular immune responses in comparison to a similar regimen that includes a vector with unmodified hexon. These differences in immunogenicity translated into a better protective efficacy against both the hepatic and red blood cell stages of P. yoelii. To our knowledge, this is the first time that a hexon modification is used to deliver a promiscuous T cell epitope. Our data support the use of such modification to enhance the immunogenicity and protective

  17. [Experimental study of human umbilical cord blood derived stromal cells transfected with recombinant adenoviral vector co-expressing VCAM-1 and GFP].

    PubMed

    Zhang, Xi; Si, Ying-Jian; Chen, Xing-Hua; Liu, Yao; Gao, Li; Gao, Lei; Peng, Xian-Gui; Wang, Qing-Yu

    2008-06-01

    This study was aimed to investigate the effect of vcam-1 gene-modified human umbilical cord blood derived stromal cells (CBDSCs) on hematopoietic regulation so as to establish the experimental foundation for further study. The target gene vcam-1 was cloned into the shuttle plasmid with the report gene GFP. The recombinant shuttle plasmid was transformed into BJ5183 bacteria to recombine with backbone vector pAdeasy-l, and the recombinant adenoviral vector ad-vcam-1-gfp was confirmed after transfection with CBDSCs. The results indicated that two fragments of about 9 kb and 2 kb were obtained after digestion of recombinant plasmid pAdTrack-vcam-1 with NotIand XhoI, and single fragment of 600 bp was obtained after amplification with PCR; two fragments of about 31 kb and 4 kb were obtained after digestion of recombinant plasmid pad-vcam-1-gfp with PacI, which suggested a successful homologous recombination. The expression of vcam-1 gene in ad-vcam-1-gfp transfected CBDSCs could be detected by immunocytochemistry, RT-PCR and fluorescent microscopy. It is concluded that the recombinant adenoviral vector ad-vcam-1-gfp has been constructed successfully, and the expression of vcam-1 is up-regulated in CBDSCs transfected by gene ad-vcam-1-gfp.

  18. Protein kinase C (PKC) phosphorylates human platelet inositol trisphosphate 5/sup +/-/-phosphomonoesterase (IP/sub 3/ 5'-p'tase) increasing phosphatase activity

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

    Connolly, T.M.; Majerus, P.W.

    1986-05-01

    Phosphoinositide breakdown in response to thrombin stimulation of human platelets generates messenger molecules that activate PKC (diglyceride) and mobilize Ca/sup + +/ (inositol tris-phosphates). The water soluble products of phospholipase C-mediated metabolism of phosphatidylinositol 4,5-diphosphate are inositol 1,4,5 P/sub 3/ (IP/sub 3/) and inositol 1:2-cyclic 4,5 P/sub 3/ (cIP/sub 3/). A specific phosphatase, IP/sub 3/ 5'-p'tase, cleaves the 5 phosphate from IP/sub 3/ or cIP/sub 3/ to form IP/sub 2/ or cIP/sub 2/ and P/sub i/, none of which mobilizes Ca/sup + +/. Thus, the IP/sub 3/ 5'-p'tase may regulate cellular responses to IP/sub 3/ or cIP/sub 3/. The authorsmore » find that IP/sub 3/ 5'-p'tase isolated from human platelets is phosphorylated by rat brain PKC, resulting in a 4-fold increase in IP/sub 3/ 5'-p'tase activity. The authors phosphorylated IP/sub 3/ 5'-p'tase using ..gamma.. /sup 32/P-ATP and found that the labeled enzyme comigrated on SDS-PAGE with the previously described 40K protein phosphorylated in response to thrombin stimulation of platelets. The similarity of the PKC-phosphorylated IP/sub 3/ 5'-p'tase observed in vitro and the thrombin-stimulated phosphorylated 40K protein known to be phosphorylated by PKC in vivo, suggests that these proteins may be the same. These results suggest that platelet Ca/sup + +/ mobilization maybe regulated by PKC phosphorylation of the IP/sub 3/ 5'-p'tase and can explain the observation that phorbol ester treatment of intact human platelets results in decreased production of IP/sub 3/ and decreased Ca/sup + +/ mobilization upon subsequent thrombin addition.« less

  19. Rietveld refinement, electronic structure and ionic conductivity of Sr{sub 4}La{sub 6}(SiO{sub 4}){sub 6}F{sub 2} and Sr{sub 4}La{sub 6}(SiO{sub 4}){sub 6}O ceramics

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

    Boughzala, Khaled, E-mail: khaledboughzala@gmail.com; Preparatory Institute for Engineering Studies, 5000 Monastir; Debbichi, Mourad

    In this paper, we report the effect of the tunnel anions on the ionic conductivity of Strontium-Lanthanum silicate apatites. The Sr{sub 4}La{sub 6}(SiO{sub 4}){sub 6}F{sub 2} and Sr{sub 4}La{sub 6}(SiO{sub 4}){sub 6}O ceramics were prepared by the solid state reaction method. X-ray diffraction, NMR spectroscopy and Raman measurements were performed to investigate the crystal structure and vibrational active modes. Moreover, the electronic structures of the crystals were evaluated by the first-principles quantum mechanical calculation based on the density functional theory. Finally, the ionic conductivity was studied according to the complex impedance method. - Graphical abstract: The relaxed primitive unit cellmore » for Sr{sub 4}La{sub 6}Fap. Display Omitted.« less

  20. Imaging analysis of nuclear antiviral factors through direct detection of incoming adenovirus genome complexes

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

    Komatsu, Tetsuro; Department of Infection Biology, Faculty of Medicine, University of Tsukuba, Tsukuba 305-8575; Will, Hans

    2016-04-22

    Recent studies involving several viral systems have highlighted the importance of cellular intrinsic defense mechanisms through nuclear antiviral proteins that restrict viral propagation. These factors include among others components of PML nuclear bodies, the nuclear DNA sensor IFI16, and a potential restriction factor PHF13/SPOC1. For several nuclear replicating DNA viruses, it was shown that these factors sense and target viral genomes immediately upon nuclear import. In contrast to the anticipated view, we recently found that incoming adenoviral genomes are not targeted by PML nuclear bodies. Here we further explored cellular responses against adenoviral infection by focusing on specific conditions asmore » well as additional nuclear antiviral factors. In line with our previous findings, we show that neither interferon treatment nor the use of specific isoforms of PML nuclear body components results in co-localization between incoming adenoviral genomes and the subnuclear domains. Furthermore, our imaging analyses indicated that neither IFI16 nor PHF13/SPOC1 are likely to target incoming adenoviral genomes. Thus our findings suggest that incoming adenoviral genomes may be able to escape from a large repertoire of nuclear antiviral mechanisms, providing a rationale for the efficient initiation of lytic replication cycle. - Highlights: • Host nuclear antiviral factors were analyzed upon adenovirus genome delivery. • Interferon treatments fail to permit PML nuclear bodies to target adenoviral genomes. • Neither Sp100A nor B targets adenoviral genomes despite potentially opposite roles. • The nuclear DNA sensor IFI16 does not target incoming adenoviral genomes. • PHF13/SPOC1 targets neither incoming adenoviral genomes nor genome-bound protein VII.« less

  1. Synthesis and crystal structure of the [Co{sub 2}(Nicotinamide){sub 4}(C{sub 4}H{sub 9}COO){sub 4}(H{sub 2}O)] complex

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

    Sadikov, G. G., E-mail: sadgg@igic.ras.ru; Antsyshkina, A. S.; Koksharova, T. V.

    2007-09-15

    The [Co{sub 2}L{sub 4}(C{sub 4}H{sub 9}COO){sub 4}(H{sub 2}O)] coordination compound of cobalt(II) valerate with nicotinamide (L) is synthesized and studied by IR spectroscopy. The crystal structure of the synthesized compound is determined. The crystals are triclinic, and the unit cell parameters are as follows: a = 10.2759(10) A, b = 16.3858(10) A, c = 16.4262(10) A, {alpha} = 100.538(10) deg., {beta} = 101.199(10) deg., {gamma} = 90.813 (10) deg., Z = 2, and space group P1-bar. The structural units of the crystal are dimeric molecular complexes in which pairs of cobalt atoms are linked by triple bridges formed by oxygenmore » atoms of two bidentately coordinated valerate anions and a water molecule. The octahedral coordination of each cobalt atom is complemented by the pyridine nitrogen atoms of two nicotinamide ligands and the oxygen atom of the monodentate valerate group. The hydrocarbon chains of the valerate anions are disordered over two or three positions each.« less

  2. Targeted Adenoviral Vector Demonstrates Enhanced Efficacy for In Vivo Gene Therapy of Uterine Leiomyoma.

    PubMed

    Abdelaziz, Mohamed; Sherif, Lotfy; ElKhiary, Mostafa; Nair, Sanjeeta; Shalaby, Shahinaz; Mohamed, Sara; Eziba, Noura; El-Lakany, Mohamed; Curiel, David; Ismail, Nahed; Diamond, Michael P; Al-Hendy, Ayman

    2016-04-01

    Gene therapy is a potentially effective non-surgical approach for the treatment of uterine leiomyoma. We demonstrated that targeted adenovirus vector, Ad-SSTR-RGD-TK/GCV, was highly effective in selectively inducing apoptosis and inhibiting proliferation of human leiomyoma cells in vitro while sparing normal myometrial cells. An in-vivo study, to compare efficacy and safety of modified adenovirus vector Ad-SSTR-RGD-TK/GCV versus untargeted vector for treatment of leiomyoma. Female nude mice were implanted with rat leiomyoma cells subcutaneously. Then mice were randomized into three groups. Group 1 received Ad-LacZ (marker gene), Group 2 received untargeted Ad-TK, and Group 3 received the targeted Ad-SSTR-RGD-TK. Tumors were measured weekly for 4 weeks. Then mice were sacrificed and tissue samples were collected. Evaluation of markers of apoptosis, proliferation, extracellular matrix, and angiogenesis was performed using Western Blot & Immunohistochemistry. Statistical analysis was done using ANOVA. Dissemination of adenovirus was assessed by PCR. In comparison with the untargeted vector, the targeted adenoviral vector significantly shrank leiomyoma size (P < 0.05), reduced expression of proliferation marker (PCNA) (P < 0.05), induced expression of apoptotic protein, c-PARP-1, (P < 0.05) and inhibited expression of extracellular matrix-related genes (TGF beta 3) and angiogenesis-related genes (VEGF & IGF-1) (P < 0.01). There were no detectable adenovirus in tested tissues other than leiomyoma lesions with both targeted and untargeted adenovirus. Targeted adenovirus, effectively reduces tumor size in leiomyoma without dissemination to other organs. Further evaluation of this localized targeted strategy for gene therapy is needed in appropriate preclinical humanoid animal models in preparation for a future pilot human trial. © The Author(s) 2016.

  3. NRfamPred: a proteome-scale two level method for prediction of nuclear receptor proteins and their sub-families.

    PubMed

    Kumar, Ravindra; Kumari, Bandana; Srivastava, Abhishikha; Kumar, Manish

    2014-10-29

    Nuclear receptor proteins (NRP) are transcription factor that regulate many vital cellular processes in animal cells. NRPs form a super-family of phylogenetically related proteins and divided into different sub-families on the basis of ligand characteristics and their functions. In the post-genomic era, when new proteins are being added to the database in a high-throughput mode, it becomes imperative to identify new NRPs using information from amino acid sequence alone. In this study we report a SVM based two level prediction systems, NRfamPred, using dipeptide composition of proteins as input. At the 1st level, NRfamPred screens whether the query protein is NRP or non-NRP; if the query protein belongs to NRP class, prediction moves to 2nd level and predicts the sub-family. Using leave-one-out cross-validation, we were able to achieve an overall accuracy of 97.88% at the 1st level and an overall accuracy of 98.11% at the 2nd level with dipeptide composition. Benchmarking on independent datasets showed that NRfamPred had comparable accuracy to other existing methods, developed on the same dataset. Our method predicted the existence of 76 NRPs in the human proteome, out of which 14 are novel NRPs. NRfamPred also predicted the sub-families of these 14 NRPs.

  4. Novel approach to abuse the hyperactive K-Ras pathway for adenoviral gene therapy of colorectal cancer

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

    Naumov, Inna; Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv; Kazanov, Dina

    2012-01-15

    Background: Functional activation of oncogenic K-Ras signaling pathway plays an important role in the early events of colorectal carcinogenesis (CRC). K-Ras proto-oncogene is involved in 35-40% of CRC cases. Mutations in the Ras gene trigger the transduction of proliferative and anti-apoptotic signals, even in the absence of extra cellular stimuli. The objective of the current study was to use a gene-targeting approach to kill human CRC cells selectively harboring mutated K-Ras. Results: A recombinant adenovirus that carries a lethal gene, PUMA, under the control of a Ras responsive promoter (Ad-Py4-SV40-PUMA) was used selectively to target CRC cells (HCT116, SW480, DLD1more » and RIE-Ras) that possess a hyperactive Ras pathway while using HT29 and RIE cells as a control that harbors wild type Ras and exhibit very low Ras activity. Control vector, without the Ras responsive promoter elements was used to assess the specificity of our 'gene therapy' approach. Both adenoviral vectors were assed in vitro and in xenograft model in vivo. Ad-Py4-SV40-PUMA showed high potency to induce {approx} 50% apoptosis in vitro, to abolish completely tumor formation by infecting cells with the Ad-Py4-SV40-PUMA prior xenografting them in nude mice and high ability to suppress by {approx} 35% tumor progression in vivo in already established tumors. Conclusions: Selective targeting of CRC cells with the activated Ras pathway may be a novel and effective therapy in CRC. The high potency of this adenoviral vector may help to overcome an undetectable micro metastasis that is the major hurdle in challenging with CRC.« less

  5. Developmental changes in the role of a pertussis toxin sensitive guanine nucleotide binding protein in the rat cardiac alpha sub 1 -adrenergic system

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

    Han, H.M.

    1989-01-01

    During development, the cardiac alpha{sub 1}-adrenergic chronotropic response changes from positive in the neonate to negative in the adult. This thesis examined the possibility of a developmental change in coupling of a PT-sensitive G-protein to the alpha{sub 1}-adrenergic receptor. Radioligand binding experiments performed with the iodinated alpha{sub 1}-selective radioligand ({sup 125}I)-I-2-({beta}-(4-hydroxphenyl)ethylaminomethyl)tetralone (({sup 125}I)-IBE 2254) demonstrated that the alpha{sub 1}-adrenergic receptor is coupled to a G-protein in both neonatal and adult rat hearts. However, in the neonate the alpha{sub 1}-adrenergic receptor is coupled to a PT-insensitive G-protein, whereas in the adult the alpha{sub 1}-adrenergic receptor is coupled to both a PT-insensitivemore » and a PT-sensitive G-protein. Consistent with the results from binding experiments, PT did not have any effect on the alpha{sub 1}-mediated positive chronotropic response in the neonate, whereas in the adult the alpha{sub 1}-mediated negative chronotropic response was completely converted to a positive one after PT-treatment. This thesis also examined the possibility of an alteration in coupling of the alpha{sub 1}-adrenergic receptor to its effector under certain circumstances such as high potassium (K{sup +}) depolarization in nerve-muscle (NM) co-cultures, a system which has been previously shown to be a convenient in vitro model to study the mature inhibitory alpha{sub 1}-response.« less

  6. United3D: a protein model quality assessment program that uses two consensus based methods.

    PubMed

    Terashi, Genki; Oosawa, Makoto; Nakamura, Yuuki; Kanou, Kazuhiko; Takeda-Shitaka, Mayuko

    2012-01-01

    In protein structure prediction, such as template-based modeling and free modeling (ab initio modeling), the step that assesses the quality of protein models is very important. We have developed a model quality assessment (QA) program United3D that uses an optimized clustering method and a simple Cα atom contact-based potential. United3D automatically estimates the quality scores (Qscore) of predicted protein models that are highly correlated with the actual quality (GDT_TS). The performance of United3D was tested in the ninth Critical Assessment of protein Structure Prediction (CASP9) experiment. In CASP9, United3D showed the lowest average loss of GDT_TS (5.3) among the QA methods participated in CASP9. This result indicates that the performance of United3D to identify the high quality models from the models predicted by CASP9 servers on 116 targets was best among the QA methods that were tested in CASP9. United3D also produced high average Pearson correlation coefficients (0.93) and acceptable Kendall rank correlation coefficients (0.68) between the Qscore and GDT_TS. This performance was competitive with the other top ranked QA methods that were tested in CASP9. These results indicate that United3D is a useful tool for selecting high quality models from many candidate model structures provided by various modeling methods. United3D will improve the accuracy of protein structure prediction.

  7. Synthesis and characterization of new fluoride-containing manganese vanadates A{sub 2}Mn{sub 2}V{sub 2}O{sub 7}F{sub 2} (A=Rb, Cs) and Mn{sub 2}VO{sub 4}F

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

    Sanjeewa, Liurukara D.; McGuire, Michael A.; Smith Pellizzeri, Tiffany M.

    2016-09-15

    Large single crystals of A{sub 2}Mn{sub 2}V{sub 2}O{sub 7}F{sub 2} (A=Rb, Cs) and Mn{sub 2}VO{sub 4}F were grown using a high-temperature (~600 °C) hydrothermal technique. Single crystal X-ray diffraction and powder X-ray diffraction were utilized to characterize the structures, which both possess MnO{sub 4}F{sub 2} building blocks. The A{sub 2}Mn{sub 2}V{sub 2}O{sub 7}F{sub 2} series crystallizes as a new structure type in space group Pbcn (No. 60), Z=4 (Rb{sub 2}Mn{sub 2}V{sub 2}O{sub 7}F{sub 2}: a=7.4389(17) Å, b=11.574(3) Å, c=10.914(2) Å; Cs{sub 2}Mn{sub 2}V{sub 2}O{sub 7}F{sub 2}: a=7.5615(15) Å, b=11.745(2) Å, c=11.127(2) Å). The structure is composed of zigzag chains ofmore » edge-sharing MnO{sub 4}F{sub 2} units running along the a-axis, and interconnected through V{sub 2}O{sub 7} pyrovanadate groups. Temperature dependent magnetic susceptibility measurements on this interesting one-dimensional structural feature based on Mn{sup 2+} indicated that Cs{sub 2}Mn{sub 2}V{sub 2}O{sub 7}F{sub 2} is antiferromagnetic with a Neél temperature, T{sub N}=~3 K and a Weiss constant, θ, of −11.7(1) K. Raman and infrared spectra were also analyzed to identify the fundamental V–O vibrational modes in Cs{sub 2}Mn{sub 2}V{sub 2}O{sub 7}F{sub 2}. Mn{sub 2}(VO{sub 4})F crystalizes in the monoclinic space group of C2/c (no. 15), Z=8 with unit cell parameters of a=13.559(2) Å, b=6.8036(7) Å, c=10.1408(13) Å and β=116.16(3)°. The structure is associated with those of triplite and wagnerite. Dynamic fluorine disorder gives rise to complex alternating chains of five-and six-coordinate Mn{sup 2+}. These interpenetrating chains are additionally connected through isolated VO{sub 4} tetrahedra to form the condensed structure. - Graphical abstract: New vanadate fluorides A{sub 2}Mn{sub 2}V{sub 2}O{sub 7}F{sub 2} (A=Rb, Cs) and Mn{sub 2}(VO{sub 4})F have been synthesized hydrothermally. Upon cooling, the one-dimensional Mn(II) substructure results in

  8. The new high-pressure borate Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O-Formation of edge-sharing BO{sub 4} tetrahedra in a hydrated borate

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

    Neumair, Stephanie C.; Kaindl, Reinhard; Huppertz, Hubert, E-mail: hubert.huppertz@uibk.ac.at

    2012-01-15

    The new borate hydrate Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O was synthesized under high-pressure/high-temperature conditions of 6 GPa and 880 Degree-Sign C in a Walker-type multianvil apparatus. The compound crystallizes in the orthorhombic space group Pbam (Z=2) with the lattice parameters a=819.0(2), b=2016.9(4), c=769.9(2) pm, V=1.2717(4) nm{sup 3}, R{sub 1}=0.0758, wR{sub 2}=0.0836 (all data). The new structure type of Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O is built up from corner-sharing BO{sub 4} tetrahedra forming corrugated layers, that are interconnected among each other by two edge-sharing BO{sub 4} tetrahedra (B{sub 2}O{sub 6} units) forming Z-shaped channels. Interestingly, the heremore » presented structure of Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O is closely related to the structures of M{sub 6}B{sub 22}O{sub 39}{center_dot}H{sub 2}O (M=Fe, Co), which exhibit BO{sub 4} tetrahedra in an intermediate state on the way to edge-sharing BO{sub 4} tetrahedra. - Graphical Abstract: The new high-pressure borate hydrate Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O is built up from corner-sharing BO{sub 4} tetrahedra forming corrugated layers, that are interconnected among each other by two edge-sharing BO{sub 4} tetrahedra (B{sub 2}O{sub 6} units). In this paper we report on synthesis, structural details, and properties of the new compound Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O. Highlights: Black-Right-Pointing-Pointer High-pressure/high-temperature synthesis of the new borate hydrate Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O. Black-Right-Pointing-Pointer In the structure of Co{sub 7}B{sub 24}O{sub 42}(OH){sub 2}{center_dot}2 H{sub 2}O, two B{sub 2}O{sub 6} units are connected to 'vierer' rings. Black-Right-Pointing-Pointer Pressure favours the formation of edge-sharing BO{sub 4} tetrahedra in the chemistry of borates.« less

  9. Sub-orbital commercial Human space flight and informed consent in the United States

    NASA Astrophysics Data System (ADS)

    Carminati, Maria-Vittoria « Giugi »; Griffith, Doug; Campbell, Mark R.

    2013-12-01

    Commercial space flight is expected to rapidly develop in the near future. This will begin with sub-orbital missions and then progress to orbital flights. In the United States, technical informed consent of space flight participants is required by the commercial space flight operator for regulatory purposes. Additionally, though not required by U.S. regulation, the aerospace medicine professional involved in the medical screening of both space flight participants and crewmembers will be asked to assist operators in obtaining medical informed consent for liability purposes. The various US federal and state regulations regarding informed consent for sub-orbital commercial space flight are evolving and are unfamiliar to most aerospace medical professionals and are reviewed and discussed.

  10. Adenoviral overexpression of Lhx2 attenuates cell viability but does not preserve the stem cell like phenotype of hepatic stellate cells.

    PubMed

    Genz, Berit; Thomas, Maria; Pützer, Brigitte M; Siatkowski, Marcin; Fuellen, Georg; Vollmar, Brigitte; Abshagen, Kerstin

    2014-11-01

    Hepatic stellate cells (HSC) are well known initiators of hepatic fibrosis. After liver cell damage, HSC transdifferentiate into proliferative myofibroblasts, representing the major source of extracellular matrix in the fibrotic organ. Recent studies also demonstrate a role of HSC as progenitor or stem cell like cells in liver regeneration. Lhx2 is described as stem cell maintaining factor in different organs and as an inhibitory transcription factor in HSC activation. Here we examined whether a continuous expression of Lhx2 in HSC could attenuate their activation and whether Lhx2 could serve as a potential target for antifibrotic gene therapy. Therefore, we evaluated an adenoviral mediated overexpression of Lhx2 in primary HSC and investigated mRNA expression patterns by qRT-PCR as well as the activation status by different in vitro assays. HSC revealed a marked increase in activation markers like smooth muscle actin alpha (αSMA) and collagen 1α independent from adenoviral transduction. Lhx2 overexpression resulted in attenuated cell viability as shown by a slightly hampered migratory and contractile phenotype of HSC. Expression of stem cell factors or signaling components was also unaffected by Lhx2. Summarizing these results, we found no antifibrotic or stem cell maintaining effect of Lhx2 overexpression in primary HSC. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. From {sub {infinity}}{sup 1}[(UO{sub 2}){sub 2}O(MoO{sub 4}){sub 4}]{sup 6-} to {sub {infinity}}{sup 1}[(UO{sub 2}){sub 2}(MoO{sub 4}){sub 3}(MoO{sub 5})]{sup 6-} infinite chains in A{sub 6}U{sub 2}Mo{sub 4}O{sub 21} (A=Na, K, Rb, Cs) compounds: Synthesis and crystal structure of Cs{sub 6}[(UO{sub 2}){sub 2}(MoO{sub 4}){sub 3}(MoO{sub 5})

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

    Yagoubi, S.; Groupe de Radiochimie, Institut de Physique Nucleaire d'Orsay, Universite Paris-Sud XI, 91406 Orsay Cedex; Obbade, S., E-mail: said.obbade@phelma.grenoble-inp.f

    2011-05-15

    A new caesium uranyl molybdate belonging to the M{sub 6}U{sub 2}Mo{sub 4}O{sub 21} family has been synthesized by solid-state reaction and its structure determined from single-crystal X-ray diffraction data. Contrary to the other alkali uranyl molybdates of this family (A=Na, K, Rb) where molybdenum atoms adopt only tetrahedral coordination and which can be formulated A{sub 6}[(UO{sub 2}){sub 2}O(MoO{sub 4}){sub 4}], the caesium compound Cs{sub 6}U{sub 2}Mo{sub 4}O{sub 21} should be written Cs{sub 6}[(UO{sub 2}){sub 2}(MoO{sub 4}){sub 3}(MoO{sub 5})] with molybdenum atoms in tetrahedral and square pyramidal environments. Cs{sub 6}[(UO{sub 2}){sub 2}(MoO{sub 4}){sub 3}(MoO{sub 5})] crystallizes in the triclinic symmetry withmore » space group P1-bar and a=10.4275(14) A, b=15.075(2) A, c=17.806(2) A, {alpha}=70.72(1){sup o}, {beta}=80.38(1){sup o} and {gamma}=86.39(1){sup o}, V=2604.7(6) A{sup 3}, Z=4, {rho}{sub mes}=5.02(2) g/cm{sup 3} and {rho}{sub cal}=5.08(3) g/cm{sup 3}. A full-matrix least-squares refinement on the basis of F{sup 2} yielded R{sub 1}=0.0464 and wR{sub 2}=0.0950 for 596 parameters with 6964 independent reflections with I{>=}2{sigma}(I) collected on a BRUKER AXS diffractometer with Mo(K{alpha}) radiation and a CCD detector. The crystal structure of Cs compound is characterized by {sub {infinity}}{sup 1}[(UO{sub 2}){sub 2}(MoO{sub 4}){sub 3}(MoO{sub 5})]{sup 6-} parallels chains built from U{sub 2}O{sub 13} dimeric units, MoO{sub 4} tetrahedra and MoO{sub 5} square pyramids, whereas, Na, K and Rb compounds are characterized by {sub {infinity}}{sup 1}[(UO{sub 2}){sub 2}O(MoO{sub 4}){sub 4}]{sup 6-} parallel chains formulated simply of U{sub 2}O{sub 13} units and MoO{sub 4} tetrahedra. Infrared spectroscopy measurements using powdered samples synthesized by solid-state reaction, confirm the structural results. The thermal stability and the electrical conductivity are also studied. The four compounds decompose at low temperature (between 540 and 610

  12. Crystallization and preliminary X-ray crystallographic studies of Drep-3, a DFF-related protein from Drosophila melanogaster

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

    Park, Hyun Ho; Tookes, Hansel Emory; Wu, Hao, E-mail: haowu@med.cornell.edu

    2006-06-01

    The D. melanogaster Drep-3 protein has been crystallized. Crystals were obtained at 293 K that diffracted to 2.8 Å resolution and belonged to space group P2{sub 1}2{sub 1}2{sub 1}. During apoptosis, DNA fragmentation is mainly mediated by the caspase-activated DFF40 nuclease. DFF40 exists as a heterodimeric complex with its inhibitor DFF45. Upon apoptosis induction, DFF45 is cleaved by caspases to allow DFF40 activation. Drep-3 is a recently identified regulator of the DFF40 system in Drosophila melanogaster. Here, Drep-3 was expressed with a C-terminal His tag in Escherichia coli and the protein was purified to homogeneity. Multi-angle light-scattering analysis showed thatmore » Drep-3 is a homotetramer in solution. Native and selenomethionine-substituted Drep-3 proteins were crystallized at 293 K and X-ray diffraction data were collected to 2.8 and 3.0 Å resolution, respectively. The crystals belong to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 56.9, b = 125.4, c = 168.7 Å. The asymmetric unit is estimated to contain one homotetramer.« less

  13. cap alpha. /sub i/-3 cDNA encodes the. cap alpha. subunit of G/sub k/, the stimulatory G protein of receptor-regulated K/sup +/ channels

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

    Codina, J.; Olate, J.; Abramowitz, J.

    1988-05-15

    cDNA cloning has identified the presence in the human genome of three genes encoding ..cap alpha.. subunits of pertussis toxin substrates, generically called G/sub i/. They are named ..cap alpha../sub i/-1, ..cap alpha../sub i/-2 and ..cap alpha../sub i/-3. However, none of these genes has been functionally identified with any of the ..cap alpha.. subunits of several possible G proteins, including pertussis toxin-sensitive G/sub p/'s, stimulatory to phospholipase C or A/sub 2/, G/sub i/, inhibitory to adenylyl cyclase, or G/sub k/, stimulatory to a type of K/sup +/ channels. The authors now report the nucleotide sequence and the complete predicted aminomore » acid sequence of human liver ..cap alpha../sub i/-3 and the partial amino acid sequence of proteolytic fragments of the ..cap alpha.. subunit of human erythrocyte G/sub k/. The amino acid sequence of the proteolytic fragment is uniquely encoded by the cDNA of ..cap alpha../sub i/-3, thus identifying it as ..cap alpha../sub k/. The probable identity of ..cap alpha../sub i/-1 with ..cap alpha../sub p/ and possible roles for ..cap alpha../sub i/-2, as well as additional roles for ..cap alpha../sub i/-1 and ..cap alpha../sub i/-3 (..cap alpha../sub k/) are discussed.« less

  14. Burn care delivery in a sub-saharan african unit: A cost analysis study.

    PubMed

    Gallaher, Jared R; Mjuweni, Stephen; Cairns, Bruce A; Charles, Anthony G

    2015-07-01

    There are significant resource challenges to burn surgical care delivery in low and middle-income countries at baseline and only a few burn cost analysis studies from sub-Saharan Africa have been performed. This is a retrospective database analysis of prospectively collected data from all patients recorded in the burn registry between June 2011 and August 2014 located at the Kamuzu Central Hospital Burn Unit in Lilongwe, Malawi. We utilized activity-based costing, a bottom-up cost analysis methodology with cost allocation that allows determination of unit cost or cost per service. 905 patients were admitted to the burn unit during the study period. The calculated total monthly burn expenditure for all cost centers was $11,622.66. Per day, the total unit cost was $387.42 with a mean daily per-patient cost of $24.26 (SD ± $6.44). Consequently, the mean cost per in-patient admission was $559.85 (SD ± $736.17). The mean daily cost per 1% total burn surface per patient at our center is $2.65 (SD ± $3.01). This burn care cost analysis study helps quantify the relative contribution of differing cost centers that comprise burn care delivery and hospital costs in a sub-Saharan African setting. Accurate and relevant cost information on hospital services at the patient level is therefore fundamental for policy makers, payers, and hospitals. Our study has demonstrated that comprehensive burn care is possible at a cost much lower than found in other burn centers in low or middle-income countries and can be sustained with moderate funding. Copyright © 2015 IJS Publishing Group Limited. Published by Elsevier Ltd. All rights reserved.

  15. Synthesis of the new quaternary sulfides K[sub 2]Y[sub 4]Sn[sub 2]S[sub 11] and BaLnAgS[sub 3] (Ln = Er, Y, Gd) and the structures of K[sub 2]Y[sub 4]Sn[sub 2]S[sub 11] and BaErAgS[sub 3

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

    Wu, Ping; Ibers, J.A.

    1994-05-01

    Several new quarternary sulfides, K[sub 2]Y[sub 4]Sn[sub 2]S[sub 11] and BaLnAgS[sub 3] (Ln = Er, Y, Gd), have been synthesized by the reaction of the constituent binary chalcogenides and elements at 1000[degrees]C. The crystal structures of K[sub 2]Y[sub 4]Sn[sub 2]S[sub 11] and BaErAgS[sub 3] have been determined by single-crystal X-ray diffraction techniques. Crystal data: K[sub 2]Y[sub 4]Sn[sub 2]S[sub 11]-space group D[sup 8][sub 4h] - P4/ncc, M = 1023.88, Z = 4, a = 8.587(1), c = 27.892(4) [angstrom] (T = 115 K), V = 2056.7(4) [angstrom][sup 3], R[sub W](F[sup 2]) = 0.093 for 1965 observations having F[sup 2][sub 0] >more » 2[sigma](F[sup 2][sub 0]); BaEr AgS[sub 3]-space group C[sup 3][sub 2H] - C2/m, M = 508.65, Z = 4, a = 17.340(4), b = 4.014(1), x = 8.509(2) [angstrom], [beta] = 103.23(3)[degrees], (T = 115 K), V = 576.5(2) [angstrom][sup 3], R[sub W](F[sup 2]) = 0.049 for 1404 observations and 48 variables, R(F) = 0.018 for 1299 observations having F[sup 2][sub 0] > 2[sigma](F[sup 2][sub 0]). In both structures, the rare-earth atoms have octahedral coordination and the octahedra form slabs through edge- and corner-sharing. These slabs are separated by K[sup +] Ba[sup 2+] cations, and are crosslinked into three-dimensional frameworks by Sn[sub 2]S[sub 6] units as edge-sharing SnS[sub 4] tetrahedral pairs in K[sub 2]Y[sub 4]Sn[sub 2]S[sub 11], and by Ag[sub 2]S[sub 9] units as corner-sharing trigonal-bipyramidal AgS[sub 5] pairs in BaEr AgS[sub 3]. From their powder diffraction patterns, BaYAgS[sub 3] and Ba GdAgS[sub 3] appear to be isostructural with BaErAgS[sub 3].« less

  16. Adenoviral Vectors Armed with Cell Fusion-Inducing Proteins as Anti-Cancer Agents

    PubMed Central

    Del Papa, Joshua; Parks, Robin J.

    2017-01-01

    Cancer is a devastating disease that affects millions of patients every year, and causes an enormous economic burden on the health care system and emotional burden on affected families. The first line of defense against solid tumors is usually extraction of the tumor, when possible, by surgical methods. In cases where solid tumors can not be safely removed, chemotherapy is often the first line of treatment. As metastatic cancers often become vigorously resistant to treatments, the development of novel, more potent and selective anti-cancer strategies is of great importance. Adenovirus (Ad) is the most commonly used virus in cancer clinical trials, however, regardless of the nature of the Ad-based therapeutic, complete responses to treatment remain rare. A number of pre-clinical studies have shown that, for all vector systems, viral spread throughout the tumor mass can be a major limiting factor for complete tumor elimination. By expressing exogenous cell-fusion proteins, many groups have shown improved spread of Ad-based vectors. This review summarizes the research done to examine the potency of Ad vectors expressing fusogenic proteins as anti-cancer therapeutics. PMID:28106842

  17. Targeted Adenoviral Vector Demonstrates Enhanced Efficacy for In Vivo Gene Therapy of Uterine Leiomyoma

    PubMed Central

    Abdelaziz, Mohamed; Sherif, Lotfy; ElKhiary, Mostafa; Nair, Sanjeeta; Shalaby, Shahinaz; Mohamed, Sara; Eziba, Noura; El-Lakany, Mohamed; Curiel, David; Ismail, Nahed; Diamond, Michael P.; Al-Hendy, Ayman

    2016-01-01

    Background: Gene therapy is a potentially effective non-surgical approach for the treatment of uterine leiomyoma. We demonstrated that targeted adenovirus vector, Ad-SSTR-RGD-TK/GCV, was highly effective in selectively inducing apoptosis and inhibiting proliferation of human leiomyoma cells in vitro while sparing normal myometrial cells. Study design: An in-vivo study, to compare efficacy and safety of modified adenovirus vector Ad-SSTR-RGD-TK/GCV versus untargeted vector for treatment of leiomyoma. Materials and methods: Female nude mice were implanted with rat leiomyoma cells subcutaneously. Then mice were randomized into three groups. Group 1 received Ad-LacZ (marker gene), Group 2 received untargeted Ad-TK, and Group 3 received the targeted Ad-SSTR-RGD-TK. Tumors were measured weekly for 4 weeks. Then mice were sacrificed and tissue samples were collected. Evaluation of markers of apoptosis, proliferation, extracellular matrix, and angiogenesis was performed using Western Blot & Immunohistochemistry. Statistical analysis was done using ANOVA. Dissemination of adenovirus was assessed by PCR. Results: In comparison with the untargeted vector, the targeted adenoviral vector significantly shrank leiomyoma size (P < 0.05), reduced expression of proliferation marker (PCNA) (P < 0.05), induced expression of apoptotic protein, c-PARP-1, (P < 0.05) and inhibited expression of extracellular matrix-related genes (TGF beta 3) and angiogenesis-related genes (VEGF & IGF-1) (P < 0.01). There were no detectable adenovirus in tested tissues other than leiomyoma lesions with both targeted and untargeted adenovirus. Conclusion: Targeted adenovirus, effectively reduces tumor size in leiomyoma without dissemination to other organs. Further evaluation of this localized targeted strategy for gene therapy is needed in appropriate preclinical humanoid animal models in preparation for a future pilot human trial. PMID:26884457

  18. Purification, identification and preliminary crystallographic studies of an allergenic protein from Lathyrus sativus

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

    Qureshi, Insaf A.; Sethi, Dhruv K.; Salunke, Dinakar M., E-mail: dinakar@nii.res.in

    2006-09-01

    A 24 kDa protein was purified from the seeds of L. sativus by ammonium sulfate fractionation and ion-exchange chromatography. Crystals were obtained by the hanging-drop vapour-diffusion method. A 24 kDa protein was purified from the seeds of Lathyrus sativus by ammonium sulfate fractionation and ion-exchange chromatography. The N-terminal amino-acid sequence showed significant homology with the 2S albumin class of seed storage proteins. The protein showed 85% sequence homology with the seed albumin of Pisum sativum within the 40 N-terminal residues. Crystals were obtained by the hanging-drop vapour-diffusion method. The crystals belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cellmore » parameters a = 43.5, b = 82.7, c = 153.4 Å.« less

  19. Lymphatic Vascular-Based Therapy for IBD

    DTIC Science & Technology

    2012-07-01

    adenoviral particles encoding VEGF -D (2X108) were injected intraperitoneally into mice to induce VEGF -D protein expression in the abdominal cavity near the...10). Expression of adenoviral proteins in mouse intestinal epithelial cells. To characterize induced VEGF expression by target tissues, VEGF -C...Mouse VEGF -D is a selective ligand for mouse VEGFR-3. Postnatally, VEGFR-3 expression is restricted to lymphatic endothelial cells [29; 30]. VEGF -D is

  20. Synthesis and structural characterization of the Zintl phases Na{sub 3}Ca{sub 3}TrPn{sub 4}, Na{sub 3}Sr{sub 3}TrPn{sub 4}, and Na{sub 3}Eu{sub 3}TrPn{sub 4} (Tr=Al, Ga, In; Pn=P, As, Sb)

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

    Wang, Yi; Suen, Nian-Tzu; College of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou 225002

    15 new quaternary Zintl phases have been synthesized by solid-state reactions from the respective elements, and their structures have been determined by single-crystal X-ray diffraction. Na{sub 3}E{sub 3}TrPn{sub 4} (E=Ca, Sr, Eu; Tr=Al, Ga, In; Pn=P, As, Sb) crystallize in the hexagonal crystal system with the non-centrosymmetric space group P6{sub 3}mc (No. 186). The structure represents a variant of the K{sub 6}HgS{sub 4} structure type (Pearson index hP22) and features [TrPn{sub 4}]{sup 9–} tetrahedral units, surrounded by Na{sup +} and Ca{sup 2+}, Sr{sup 2+}, Eu{sup 2+} cations. The nominal formula rationalization [Na{sup +}]{sub 3}[E{sup 2+}]{sub 3}[TrPn{sub 4}]{sup 9–} follows themore » octet rule, suggesting closed-shell configurations for all atoms and intrinsic semiconducting behavior. However, structure refinements for several members hint at disorder and mixing of cations that potentially counteract the optimal valence electron count. - Graphical abstract: The hexagonal, non-centrosymmetric structure of Na{sub 3}E{sub 3}TrPn{sub 4} (E=Ca, Sr, Eu; Tr=Al, Ga, In; Pn=P, As, Sb) features [TrPn{sub 4}]{sup 9–} tetrahedral units, surrounded by Na{sup +} and Ca{sup 2+}, Sr{sup 2+}, Eu{sup 2+} cations. - Highlights: • 15 quaternary phosphides, arsenides, and antimonides are synthesized and structurally characterized. • The structure is a variant of the hexagonal K{sub 6}HgS{sub 4}-type, with distinctive pattern for the cations. • Occupational and/or positional disorder of yet unknown origin exists for some members of the series.« less

  1. Electron diffraction and high-resolution electron microscopy studies on layered Li{sub 2−δ}(Mn{sub 1−x}Co{sub x}){sub 1+δ}O{sub 3}

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

    Fujii, Hiroki, E-mail: fujii.hiroki@nims.go.jp; Ozawa, Kiyoshi; Mochiku, Takashi

    2013-07-15

    The structure of Li{sub 2}MnO{sub 3}–LiCoO{sub 2} solid solutions or Li{sub 2−δ}(Mn{sub 1−x}Co{sub x}){sub 1+δ}O{sub 3} (LMCO) which are promising lithium-battery cathodes was studied by high-resolution electron microscopy (HREM). X-ray diffraction (XRD) analyses indicate that Li{sub 1.6}Mn{sub 0.2}Co{sub 1.2}O{sub 3} and Li{sub 1.7}Mn{sub 0.4}Co{sub 0.9}O{sub 3} take a rhombohedral (R3-bar m space group) structure, whereas Li{sub 1.85}Mn{sub 0.7}Co{sub 0.45}O{sub 3} and Li{sub 1.95}Mn{sub 0.9}Co{sub 0.15}O{sub 3} take a monoclinic (C2/m) superstructure. HREM studies on those samples reveal that all of the observed crystallites in sample Li{sub 1.95}Mn{sub 0.9}Co{sub 0.15}O{sub 3} contain sharp stripe structures due to the planar defects alongmore » the c{sub h}-axis (stacking direction of the hexagonal close-packed planes) of parent rhombohedral R3-bar m cell. Such stripe structures become faint with increasing x, Co content. The origin of the planar defects is mainly attributed not to the different structures but to the different local orientations of the monoclinic LMCO. Local disordering of Li and (Mn,Co) in (Li,Mn,Co) planes is developed with increasing x and the structures are transformed from C2/m to R3-bar m. The alternative (Li,Mn,Co)-plane stacking structure, trigonal P3{sub 1}12 LMCO, was occasionally observed. However, only P3{sub 1}12 LMCO platelets with a thickness of a single unit cell were detected. - Graphical abstract: An HREM image for Li{sub 1.95}(Mn{sub 0.9}Co{sub 0.15})O{sub 3} and schematic drawings of C2/m and P3{sub 1}12 Li{sub 2}MnO{sub 3} projected along the various zone axes. Each projected unit cell is indicated by rectangles and parallelograms . - Highlights: • Li{sub 2−δ}(Mn{sub 1−x}Co{sub x}){sub 1+δ}O{sub 3} was studied by high-resolution electron microscopy (HREM). • HREM studies revealed a large amount of planar defects in C2/m Li{sub 1.95}Mn{sub 0.9}Co{sub 0.15}O{sub 3}. • The origin of the defects is the

  2. Supra-domains: evolutionary units larger than single protein domains.

    PubMed

    Vogel, Christine; Berzuini, Carlo; Bashton, Matthew; Gough, Julian; Teichmann, Sarah A

    2004-02-20

    Domains are the evolutionary units that comprise proteins, and most proteins are built from more than one domain. Domains can be shuffled by recombination to create proteins with new arrangements of domains. Using structural domain assignments, we examined the combinations of domains in the proteins of 131 completely sequenced organisms. We found two-domain and three-domain combinations that recur in different protein contexts with different partner domains. The domains within these combinations have a particular functional and spatial relationship. These units are larger than individual domains and we term them "supra-domains". Amongst the supra-domains, we identified some 1400 (1203 two-domain and 166 three-domain) combinations that are statistically significantly over-represented relative to the occurrence and versatility of the individual component domains. Over one-third of all structurally assigned multi-domain proteins contain these over-represented supra-domains. This means that investigation of the structural and functional relationships of the domains forming these popular combinations would be particularly useful for an understanding of multi-domain protein function and evolution as well as for genome annotation. These and other supra-domains were analysed for their versatility, duplication, their distribution across the three kingdoms of life and their functional classes. By examining the three-dimensional structures of several examples of supra-domains in different biological processes, we identify two basic types of spatial relationships between the component domains: the combined function of the two domains is such that either the geometry of the two domains is crucial and there is a tight constraint on the interface, or the precise orientation of the domains is less important and they are spatially separate. Frequently, the role of the supra-domain becomes clear only once the three-dimensional structure is known. Since this is the case for only a

  3. Production of first generation adenoviral vectors for preclinical protocols: amplification, purification and functional titration.

    PubMed

    Armendáriz-Borunda, Juan; Bastidas-Ramírez, Blanca Estela; Sandoval-Rodríguez, Ana; González-Cuevas, Jaime; Gómez-Meda, Belinda; García-Bañuelos, Jesús

    2011-11-01

    Gene therapy represents a promising approach in the treatment of several diseases. Currently, the ideal vector has yet to be designed; though, adenoviral vectors (Ad-v) have provided the most utilized tool for gene transfer due principally to their simple production, among other specific characteristics. Ad-v viability represents a critical variable that may be affected by storage or shipping conditions and therefore it is advisable to be assessed previously to protocol performance. The present work is unique in this matter, as the complete detailed process to obtain Ad-v of preclinical grade is explained. Amplification in permissive HEK-293 cells, purification in CsCl gradients in a period of 10 h, spectrophotometric titration of viral particles (VP) and titration of infectious units (IU), yielding batches of AdβGal, AdGFP, AdHuPA and AdMMP8, of approximately 10¹³-10¹⁴ VP and 10¹²-10¹³ IU were carried out. In vivo functionality of therapeutic AdHuPA and AdMMP8 was evidenced in rats presenting CCl₄-induced fibrosis, as more than 60% of fibrosis was eliminated in livers after systemic delivery through iliac vein in comparison with irrelevant AdβGal. Time required to accomplish the whole Ad-v production steps, including IU titration was 20 to 30 days. We conclude that production of Ad-v following standard operating procedures assuring vector functionality and the possibility to effectively evaluate experimental gene therapy results, leaving aside the use of high-cost commercial kits or sophisticated instrumentation, can be performed in a conventional laboratory of cell culture. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  4. Application of CFB technology for large power generating units and CO{sub 2} capture

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

    Ryabov, G. A., E-mail: georgy.ryabov@gmail.com; Folomeev, O. M.; Sankin, D. A.

    2010-07-15

    Data on the development of the circulating fluidized bed (CFB) technology for combustion of fuels in large power generating units are examined. The problems with raising the steam parameters and unit power of boilers with a circulating fluidized bed are examined. With the boiler system at the 460 MW unit at Lagisza (Poland) as an example, the feasibility of raising the efficiency of units with CFB boilers through deep recovery of the heat of the effluent gases and reducing expenditure for in-house needs is demonstrated. Comparative estimates of the capital and operating costs of 225 and 330 MW units aremore » used to determine the conditions for optimum use of CFB boilers in the engineering renovation of thermal power plants in Russia. New areas for the application of CFB technology in CO{sub 2} capture are analyzed in connection with the problem of reducing greenhouse gas emissions.« less

  5. Design and syntheses of hybrid metal–organic materials based on K{sub 3}[M(C{sub 2}O{sub 4}){sub 3}]·3H{sub 2}O [M(III)=Fe, Al, Cr] metallotectons

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

    Sun, Yayong; Zong, Yingxia; Ma, Haoran

    2016-05-15

    By using K{sub 3}[M(C{sub 2}O{sub 4}){sub 3}]·3H{sub 2}O [M(III)=Fe, Al, Cr] (C{sub 2}O{sub 4}{sup 2−}=oxalate) metallotectons as the starting material, we have synthesized eight novel complexes with formulas [{Fe(C_2O_4)_2(H_2O)_2}{sub 2}]·(H–L{sub 1}){sub 2}·H{sub 2}O 1, [Fe(C{sub 2}O{sub 4})Cl{sub 2}]·(H{sub 2}–L{sub 2}){sub 0.5}·(L{sub 2}){sub 0.5}·H{sub 2}O 2, [{Fe(C_2O_4)_1_._5Cl_2}{sub 2}]·(H–L{sub 3}){sub 4}3, [Fe{sub 2}(C{sub 2}O{sub 4})Cl{sub 8}]·(H{sub 2}–L{sub 4}){sub 2}·2H{sub 2}O 4, K[Al(C{sub 2}O{sub 4}){sub 3}]·(H{sub 2}–L{sub 5})·2H{sub 2}O 5, K[Al(C{sub 2}O{sub 4}){sub 3}]·(H–L{sub 6}){sub 2}·2H{sub 2}O 6, K[Cr(C{sub 2}O{sub 4}){sub 3}]·2H{sub 2}O 7, Na[Fe(C{sub 2}O{sub 4}){sub 3}]·(H–L{sub 6}){sub 2}·2H{sub 2}O 8 (with L{sub 1}=4-dimethylaminopyridine, L{sub 2}=2,3,5,6-tetramethylpyrazine, L{sub 3}=2-aminobenzimidazole, L{sub 4}=1,4-bis-(1H-imidazol-1-yl)benzene, L{sub 5}=1,4-bis((2-methylimidazol-1-yl)methyl)benzene,more » L{sub 6}=2-methylbenzimidazole). Their structures have been determined by single-crystal X-ray diffraction analyses, elemental analyses, IR spectra and thermogravimetric analyses. Compound 3 is a 2D H-bonded supramolecular architecture. Others are 3D supramolecular structures. Compound 1 shows a [Fe(C{sub 2}O{sub 4}){sub 2}(H{sub 2}O){sub 2}]{sup −} unit and 3D antionic H-bonded framework. Compound 2 features a [Fe(C{sub 2}O{sub 4})Cl{sub 2}]{sup -} anion and 1D iron-oxalate-iron chain. Compound 3 features a [Fe{sub 2}(C{sub 2}O{sub 4}){sub 3}Cl{sub 4}]{sup 4−} unit. Compound 4 features distinct [Fe{sub 2}(C{sub 2}O{sub 4})Cl{sub 8}]{sup 4−} units, which are mutual linked by water molecules to generated a 2D H-bonded network. Compound 5 features infinite ladder-like chains constructed by [Al(C{sub 2}O{sub 4}){sub 3}]{sup 3−} units and K{sup +} cations. The 1D chains are further extended into 3D antionic H-bonded framework through O–H···O H-bonds. Compounds 6–8 show 2D [KAl(C{sub

  6. Hf{sub 3}Fe{sub 4}Sn{sub 4} and Hf{sub 9}Fe{sub 4−x}Sn{sub 10+x}: Two stannide intermetallics with low-dimensional iron sublattices

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

    Calta, Nicholas P.; Kanatzidis, Mercouri G., E-mail: m-kanatzidis@northwestern.edu; Materials Science Division, Argonne National Laboratory

    This article reports two new Hf-rich intermetallics synthesized using Sn flux: Hf{sub 3}Fe{sub 4}Sn{sub 4} and Hf{sub 9}Fe{sub 4−x}Sn{sub 10+x}. Hf{sub 3}Fe{sub 4}Sn{sub 4} adopts an ordered variant the Hf{sub 3}Cu{sub 8} structure type in orthorhombic space group Pnma with unit cell edges of a=8.1143(5) Å, b=8.8466(5) Å, and c=10.6069(6) Å. Hf{sub 9}Fe{sub 4−x}Sn{sub 10+x}, on the other hand, adopts a new structure type in Cmc2{sub 1} with unit cell edges of a=5.6458(3) Å, b=35.796(2) Å, and c=8.88725(9) Å for x=0. It exhibits a small amount of phase width in which Sn substitutes on one of the Fe sites. Bothmore » structures are fully three-dimensional and are characterized by pseudo one- and two-dimensional networks of Fe–Fe homoatomic bonding. Hf{sub 9}Fe{sub 4−x}Sn{sub 10+x} exhibits antiferromagnetic order at T{sub N}=46(2) K and its electrical transport behavior indicates that it is a normal metal with phonon-dictated resistivity. Hf{sub 3}Fe{sub 4}Sn{sub 4} is also an antiferromagnet with a rather high ordering temperature of T{sub N}=373(5) K. Single crystal resistivity measurements indicate that Hf{sub 3}Fe{sub 4}Sn{sub 4} behaves as a Fermi liquid at low temperatures, indicating strong electron correlation. - Graphical abstract: Slightly different growth conditions in Sn flux produce two new intermetallic compounds: Hf{sub 3}Fe{sub 4}Sn{sub 4} and Hf{sub 9}Fe{sub 4−x}Sn{sub 10+x}. - Highlights: • Single crystals of both Hf{sub 3}Fe{sub 4}Sn{sub 4} and Hf{sub 9}Fe{sub 4−x}Sn{sub 10+x} were grown using Sn flux. • The crystal structures were determined using single crystal X-ray diffraction. • The Fe moments in Hf{sub 3}Fe{sub 4}Sn{sub 4} display AFM order below T{sub N}=373 K. • The Fe moments in Hf{sub 9}Fe{sub 4−x}Sn{sub 10+x} display AFM order below T{sub N}=46 K.« less

  7. Crystal structure of Rb{sub 2}Mn{sub 3}(H{sub 2}O){sub 2}[P{sub 2}O{sub 7}]{sub 2}, a new representative of the family of hydrated diphosphates

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

    Kiriukhina, G. V., E-mail: g-biralo@yandex.ru; Yakubovich, O. V.; Dimitrova, O. V.

    2016-09-15

    The crystal structure of Rb{sub 2}Mn{sub 3}(H{sub 2}O){sub 2}[P{sub 2}O{sub 7}]{sub 2}, a new phase obtained in the form of single crystals under hydrothermal conditions in the MnCl{sub 2}–Rb{sub 3}PO{sub 4}–H{sub 2}O system, is determined by X-ray diffraction (Xcalibur-S-CCD diffractometer, R = 0.0270): a = 9.374(2), b = 8.367(2), c = 9.437(2) Å, ß = 99.12(2)°, space group P2{sub 1}/c, Z = 2, D{sub x} = 3.27 g/cm{sup 3}. A correlation between the unit-cell parameters and the size of cations forming the crystal structures of isostructural A{sub 2}M{sub 3}(H{sub 2}O){sub 2}[P{sub 2}O{sub 7}]{sub 2} diphosphates (A = K, NH{sub 4},more » Rb, or Na; {sub M} = Mn, Fe, Co, or Ni) is revealed. It is shown that, due to the topological similarity, the structures of diphosphates and orthophosphates of the farringtonite structural type can undergo mutual transformations.« less

  8. New multicell model for describing the atomic structure of La{sub 3}Ga{sub 5}SiO{sub 14} piezoelectric crystal: Unit cells of different compositions in the same single crystal

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

    Dudka, A. P., E-mail: dudka@ns.crys.ras.ru

    2017-03-15

    Accurate X-ray diffraction study of langasite (La{sub 3}Ga{sub 5}SiO{sub 14}) single crystal has been performed using the data obtained on a diffractometer equipped with a CCD area detector at 295 and 90.5 K. Within the known La{sub 3}Ga{sub 5}SiO{sub 14} model, Ga and Si cations jointly occupy the 2d site. A new model of a “multicell” consisting of two different unit cells is proposed. Gallium atoms occupy the 2d site in one of these cells, and silicon atoms occupy this site in the other cell; all other atoms correspondingly coordinate these cations. This structure implements various physical properties exhibited bymore » langasite family crystals. The conclusions are based on processing four data sets obtained with a high resolution (sin θ/λ ≤ 1.35 Å{sup –1}), the results reproduced in repeated experiments, and the high relative precision of the study (sp. gr. P321, Z = 1; at 295 K, a = 8.1652(6) Å, c = 5.0958(5) Å, R/wR = 0.68/0.68%, 3927 independent reflections; at 90.5 K, a = 8.1559(4) Å, c = 5.0913(6) Å, R/wR = 0.92/0.93%, 3928 reflections).« less

  9. Sprouty is a cytoplasmic target of adenoviral E1A oncoproteins to regulate the receptor tyrosine kinase signalling pathway

    PubMed Central

    2011-01-01

    Background Oncoproteins encoded by the early region of adenoviruses have been shown to be powerful tools to study gene regulatory mechanisms, which affect major cellular events such as proliferation, differentiation, apoptosis and oncogenic transformation. They are possesing a key role to favor viral replication via their interaction with multiple cellular proteins. In a yeast two-hybrid screen we have identified Sprouty1 (Spry1) as a target of adenoviral E1A Oncoproteins. Spry proteins are central and complex regulators of the receptor tyrosine kinase (RTK) signalling pathway. The deregulation of Spry family members is often associated with alterations of the RTK signalling and its downstream effectors, leading to the ERK pathway. Results Here, we confirm our yeast two-hybrid data, showing the interaction between Spry1 and E1A in GST pull-down and immunoprecipitation assays. We also demonstrated the interaction of E1A with two further Spry isoforms. Using deletion mutants we identified the N-terminus and the CR conserved region (CR) 3 of E1A- and the C-terminal half of Spry1, which contains the highly conserved Spry domain, as the essential sites for direct interaction between Spry and E1A. Immunofluorescent microscopy data revealed a co-localization of E1A13S with Spry1 in the cytoplasm. SRE and TRE reporter assays demonstrated that co-expression of Spry1 with E1A13S abolishes the inhibitory function of Spry1 in RTK signalling, which is consequently accompanied with a decrease of E1A13S-induced gene expression. Conclusions These results establish Spry1 as a cytoplasmic localized cellular target for E1A oncoproteins to regulate the RTK signalling pathway, and consequently cellular events downstream of RTK that are essential for viral replication and transformation. PMID:21518456

  10. Structural phase transition in deuterated benzil C{sub 14}D{sub 10}O{sub 2}: Neutron inelastic scattering

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

    Goossens, D. J.; Welberry, T. R.; Hagen, M. E.

    2006-04-01

    Neutron inelastic scattering has been used to examine the structural phase transition in deuterated benzil C{sub 14}D{sub 10}O{sub 2}. The transition in benzil, in which the unit cell goes from a trigonal P3{sub 1}21 unit cell above T{sub C} to a cell doubled P2{sub 1} unit cell below T{sub C}, leads to the emergence of a Bragg peak at the M-point of the high temperature Brillouin zone. It has previously been suggested that the softening of a transverse optic phonon at the {gamma}-point leads to the triggering of an instability at the M-point causing the transition to occur. This suggestionmore » has been investigated by measuring the phonon spectrum at the M-point for a range of temperatures above T{sub C} and the phonon dispersion relation along the {gamma}-M direction just above T{sub C}. It is found that the transverse acoustic phonon at the M-point is of lower energy than the {gamma}-point optic mode and has a softening with temperature as T approaches T{sub C} from above that is much faster than that of the {gamma}-point optic mode. This behavior is inconsistent with the view that the {gamma}-point mode is responsible for triggering the phase transition. Rather the structural phase transition in benzil appears to be driven by a conventional soft TA mode at the M-point.« less

  11. United States aid policy and induced abortion in sub-Saharan Africa.

    PubMed

    Bendavid, Eran; Avila, Patrick; Miller, Grant

    2011-12-01

    To determine whether the Mexico City Policy, a United States government policy that prohibits funding to nongovernmental organizations performing or promoting abortion, was associated with the induced abortion rate in sub-Saharan Africa. Women in 20 African countries who had induced abortions between 1994 and 2008 were identified in Demographic and Health Surveys. A country's exposure to the Mexico City Policy was considered high (or low) if its per capita assistance from the United States for family planning and reproductive health was above (or below) the median among study countries before the policy's reinstatement in 2001. Using logistic regression and a difference-in-difference design, the authors estimated the differential change in the odds of having an induced abortion among women in high exposure countries relative to low exposure countries when the policy was reinstated. The study included 261,116 women aged 15 to 44 years. A comparison of 1994-2000 with 2001-2008 revealed an adjusted odds ratio for induced abortion of 2.55 for high-exposure countries versus low-exposure countries under the policy (95% confidence interval, CI: 1.76-3.71). There was a relative decline in the use of modern contraceptives in the high-exposure countries over the same time period. The induced abortion rate in sub-Saharan Africa rose in high-exposure countries relative to low-exposure countries when the Mexico City Policy was reintroduced. Reduced financial support for family planning may have led women to substitute abortion for contraception. Regardless of one's views about abortion, the findings may have important implications for public policies governing abortion.

  12. Crystallization and preliminary crystallographic analysis of the cellulose biosynthesis-related protein CMCax from Acetobacter xylinum

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

    Kawano, Shin; Yasutake, Yoshiaki; Tajima, Kenji

    2005-02-01

    The cellulose biosynthesis-related protein CMCax from A. xylinum has been purified and crystallized. The crystals of CMCax belong to the primitive hexagonal space group P6{sub 1} or P6{sub 5}, with unit-cell parameters a = b = 89.1, c = 94.2 Å.

  13. Crystallization and initial X-ray analysis of polyhydroxyalkanoate granule-associated protein from Aeromonas hydrophila

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

    Zhao, Minglian; Li, Zhenguo; Zheng, Wei

    The phasin PhaP{sub Ah} from A. hydrophila strain 4AK4 was crystallized using the hanging-drop vapour-diffusion method. Polyhydroxyalkanoate (PHA) granule-associated proteins (phasins) were discovered in PHA-accumulating bacteria. They play a crucial role as a structural protein during initial PHA-granule formation and granule growth and also serve as interfaces for granule stabilization in vivo. The phasin PhaP{sub Ah} from Aeromonas hydrophila strain 4AK4 was crystallized using the hanging-drop vapour-diffusion method. Single crystals were cryocooled for X-ray diffraction analysis. The phasin crystals belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 80.8, b = 108.9, c = 134.4 Å.

  14. In situ reprogramming to transdifferentiate fibroblasts into cardiomyocytes using adenoviral vectors: Implications for clinical myocardial regeneration.

    PubMed

    Mathison, Megumi; Singh, Vivek P; Chiuchiolo, Maria J; Sanagasetti, Deepthi; Mao, Yun; Patel, Vivekkumar B; Yang, Jianchang; Kaminsky, Stephen M; Crystal, Ronald G; Rosengart, Todd K

    2017-02-01

    The reprogramming of cardiac fibroblasts into induced cardiomyocyte-like cells improves ventricular function in myocardial infarction models. Only integrating persistent expression vectors have thus far been used to induce reprogramming, potentially limiting its clinical applicability. We therefore tested the reprogramming potential of nonintegrating, acute expression adenoviral (Ad) vectors. Ad or lentivirus vectors encoding Gata4 (G), Mef2c (M), and Tbx5 (T) were validated in vitro. Sprague-Dawley rats then underwent coronary ligation and Ad-mediated administration of vascular endothelial growth factor to generate infarct prevascularization. Three weeks later, animals received Ad or lentivirus encoding G, M, or T (AdGMT or LentiGMT) or an equivalent dose of a null vector (n = 11, 10, and 10, respectively). Outcomes were analyzed by echocardiography, magnetic resonance imaging, and histology. Ad and lentivirus vectors provided equivalent G, M, and T expression in vitro. AdGMT and LentiGMT both likewise induced expression of the cardiomyocyte marker cardiac troponin T in approximately 6% of cardiac fibroblasts versus <1% cardiac troponin T expression in AdNull (adenoviral vector that does not encode a transgene)-treated cells. Infarcted myocardium that had been treated with AdGMT likewise demonstrated greater density of cells expressing the cardiomyocyte marker beta myosin heavy chain 7 compared with AdNull-treated animals. Echocardiography demonstrated that AdGMT and LentiGMT both increased ejection fraction compared with AdNull (AdGMT: 21% ± 3%, LentiGMT: 14% ± 5%, AdNull: -0.4% ± 2%; P < .05). Ad vectors are at least as effective as lentiviral vectors in inducing cardiac fibroblast transdifferentiation into induced cardiomyocyte-like cells and improving cardiac function in postinfarct rat hearts. Short-term expression Ad vectors may represent an important means to induce cardiac cellular reprogramming in humans. Copyright © 2016 The American

  15. Syntheses and crystal structures of two new hydrated borates, Zn{sub 8}[(BO{sub 3}){sub 3}O{sub 2}(OH){sub 3}] and Pb[B{sub 5}O{sub 8}(OH)].1.5H{sub 2}O

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

    Chen Xuean; Zhao Yinghua; Chang Xinan

    Two new hydrated borates, Zn{sub 8}[(BO{sub 3}){sub 3}O{sub 2}(OH){sub 3}] and Pb[B{sub 5}O{sub 8}(OH)].1.5H{sub 2}O, have been prepared by hydrothermal reactions at 170 {sup o}C. Single-crystal X-ray structural analyses showed that Zn{sub 8}[(BO{sub 3}){sub 3}O{sub 2}(OH){sub 3}] crystallizes in a non-centrosymmetric space group R32 with a=8.006(2) A, c=17.751(2) A, Z=3 and Pb[B{sub 5}O{sub 8}(OH)].1.5H{sub 2}O in a triclinic space group P1-bar with a=6.656(2) A, b=6.714(2) A, c=10.701(2) A, {alpha}=99.07(2){sup o}, {beta}=93.67(2){sup o}, {gamma}=118.87(1){sup o}, Z=2. Zn{sub 8}[(BO{sub 3}){sub 3}O{sub 2}(OH){sub 3}] represents a new structure type in which Zn-centered tetrahedra are connected via common vertices leading to helical ribbons {submore » {infinity}} {sup 1}[Zn{sub 8}O{sub 15}(OH){sub 3}]{sup 17-} that pack side by side and are further condensed through sharing oxygen atoms to form a three-dimensional {sub {infinity}} {sup 3}[Zn{sub 8}O{sub 11}(OH){sub 3}]{sup 9-} framework. The boron atoms are incorporated into the channels in the framework to complete the final structure. Pb[B{sub 5}O{sub 8}(OH)].1.5H{sub 2}O is a layered compound containing double ring [B{sub 5}O{sub 8}(OH)]{sup 2-} building units that share exocyclic oxygen atoms to form a two-dimensional layer. Symmetry-center-related layers are stacked along the c-axis and held together by interlayer Pb{sup 2+} ions and water molecules via electrostatic and hydrogen bonding interactions. The IR spectra further confirmed the existence of both triangular BO{sub 3} and OH groups in Zn{sub 8}[(BO{sub 3}){sub 3}O{sub 2}(OH){sub 3}], and BO{sub 3}, BO{sub 4}, OH groups as well as guest water molecules in Pb[B{sub 5}O{sub 8}(OH)].1.5H{sub 2}O. -- Zn{sub 8}[(BO{sub 3}){sub 3}O{sub 2}(OH){sub 3}] represents a new structure type in which Zn-centered tetrahedra are connected via common vertices to form a three-dimensional framework. The boron atoms are incorporated into the channels in the framework to

  16. Helper-dependent adenoviral vectors for liver-directed gene therapy of primary hyperoxaluria type 1

    PubMed Central

    Castello, Raffaele; Borzone, Roberta; D’Aria, Stefania; Annunziata, Patrizia; Piccolo, Pasquale; Brunetti-Pierri, Nicola

    2015-01-01

    Primary hyperoxaluria type 1 (PH1) is an inborn error of liver metabolism due to deficiency of the peroxisomal enzyme alanine:glyoxylate aminotransferase (AGT) which catalyzes conversion of glyoxylate into glycine. AGT deficiency results in overproduction of oxalate which ultimately leads to end-stage renal disease and death. Organ transplantation as either preemptive liver transplantation or combined liver/kidney transplantation is the only available therapy to prevent disease progression. Gene therapy is an attractive option to provide an alternative treatment for PH1. Towards this goal, we investigated helper-dependent adenoviral (HDAd) vectors for liver-directed gene therapy of PH1. Compared to saline controls, AGT-deficient mice injected with an HDAd encoding the AGT under the control of a liver-specific promoter showed a significant reduction of hyperoxaluria and less increase of urinary oxalate following challenge with Ethylene Glycol (EG), a precursor of glyoxylate. These studies may thus pave the way to clinical application of HDAd for PH1 gene therapy. PMID:26609667

  17. Purification, identification and preliminary crystallographic studies of a 2S albumin seed protein from Lens culinaris

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

    Gupta, Pankaj; Gaur, Vineet; Salunke, Dinakar M., E-mail: dinakar@nii.res.in

    2008-08-01

    A 2S albumin from L. culinaris was purified and crystallized and preliminary crystallographic studies were carried out. Lens culinaris (lentil) is a widely consumed high-protein-content leguminous crop. A 2S albumin protein (26.5 kDa) has been identified using NH{sub 2}-terminal sequencing from a 90% ammonium sulfate saturation fraction of total L. culinaris seed protein extract. The NH{sub 2}-terminal sequence shows very high homology to PA2, an allergy-related protein from Pisum sativum. The 2S albumin protein was purified using a combination of size-exclusion and ion-exchange chromatography. Crystals of the 2S seed albumin obtained using the hanging-drop vapour-diffusion method diffracted to 2.5 Åmore » resolution and were indexed in space group P4{sub 1} (or P4{sub 3}), with unit-cell parameters a = b = 78.6, c = 135.2 Å.« less

  18. Cellular recovery from exposure to sub-optimal concentrations of AB toxins that inhibit protein synthesis.

    PubMed

    Cherubin, Patrick; Quiñones, Beatriz; Teter, Ken

    2018-02-06

    Ricin, Shiga toxin, exotoxin A, and diphtheria toxin are AB-type protein toxins that act within the host cytosol and kill the host cell through pathways involving the inhibition of protein synthesis. It is thought that a single molecule of cytosolic toxin is sufficient to kill the host cell. Intoxication is therefore viewed as an irreversible process. Using flow cytometry and a fluorescent reporter system to monitor protein synthesis, we show a single molecule of cytosolic toxin is not sufficient for complete inhibition of protein synthesis or cell death. Furthermore, cells can recover from intoxication: cells with a partial loss of protein synthesis will, upon removal of the toxin, increase the level of protein production and survive the toxin challenge. Thus, in contrast to the prevailing model, ongoing toxin delivery to the cytosol appears to be required for the death of cells exposed to sub-optimal toxin concentrations.

  19. Purification, crystallization and preliminary X-ray diffraction analysis of water-soluble chlorophyll-binding protein from Chenopodium album

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

    Ohtsuki, Takayuki; Ohshima, Shigeru; Uchida, Akira, E-mail: auchida@biomol.sci.toho-u.ac.jp

    2007-09-01

    A water-soluble chlorophyll-binding protein with photoconvertibility from C. album was extracted, purified and crystallized in a darkroom. The crystal diffracted to around 2.0 Å resolution. A water-soluble chlorophyll-binding protein (WSCP) with photoconvertibility from Chenopodium album was extracted, purified and crystallized in a darkroom. Green crystals suitable for data collection appeared in about 10 d. A native data set was collected to 2.0 Å resolution at 100 K. The space group of the crystal was determined to be orthorhombic I222 or I2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 48.13, b = 60.59, c = 107.21 Å. Preliminary analysis ofmore » the X-ray data indicated that there is one molecule per asymmetric unit.« less

  20. Derivation of Design Loads and Random Vibration specifications for Spacecraft Instruments and Sub-Units

    NASA Astrophysics Data System (ADS)

    Fransen, S.; Yamawaki, T.; Akagi, H.; Eggens, M.; van Baren, C.

    2014-06-01

    After a first estimation based on statistics, the design loads for instruments are generally estimated by coupled spacecraft/instrument sine analysis once an FE-model of the spacecraft is available. When the design loads for the instrument have been derived, the next step in the process is to estimate the random vibration environment at the instrument base and to compute the RMS load at the centre of gravity of the instrument by means of vibro-acoustic analysis. Finally the design loads of the light-weight sub-units of the instrument can be estimated through random vibration analysis at instrument level, taking into account the notches required to protect the instrument interfaces in the hard- mounted random vibration test. This paper presents the aforementioned steps of instrument and sub-units loads derivation in the preliminary design phase of the spacecraft and identifies the problems that may be encountered in terms of design load consistency between low-frequency and high-frequency environments. The SpicA FAR-infrared Instrument (SAFARI) which is currently developed for the Space Infrared Telescope for Cosmology and Astrophysics (SPICA) will be used as a guiding example.

  1. United States aid policy and induced abortion in sub-Saharan Africa

    PubMed Central

    Avila, Patrick; Miller, Grant

    2011-01-01

    Abstract Objective To determine whether the Mexico City Policy, a United States government policy that prohibits funding to nongovernmental organizations performing or promoting abortion, was associated with the induced abortion rate in sub-Saharan Africa. Methods Women in 20 African countries who had induced abortions between 1994 and 2008 were identified in Demographic and Health Surveys. A country’s exposure to the Mexico City Policy was considered high (or low) if its per capita assistance from the United States for family planning and reproductive health was above (or below) the median among study countries before the policy’s reinstatement in 2001. Using logistic regression and a difference-in-difference design, the authors estimated the differential change in the odds of having an induced abortion among women in high exposure countries relative to low exposure countries when the policy was reinstated. Findings The study included 261 116 women aged 15 to 44 years. A comparison of 1994–2000 with 2001–2008 revealed an adjusted odds ratio for induced abortion of 2.55 for high-exposure countries versus low-exposure countries under the policy (95% confidence interval, CI: 1.76–3.71). There was a relative decline in the use of modern contraceptives in the high-exposure countries over the same time period. Conclusion The induced abortion rate in sub-Saharan Africa rose in high-exposure countries relative to low-exposure countries when the Mexico City Policy was reintroduced. Reduced financial support for family planning may have led women to substitute abortion for contraception. Regardless of one’s views about abortion, the findings may have important implications for public policies governing abortion. PMID:22271944

  2. Structure, microstructure and infrared studies of Ba{sub 0.06}(Na{sub 1/2}Bi{sub 1/2}){sub 0.94}TiO{sub 3}-NaNbO{sub 3} ceramics

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

    Roy, Sumit K., E-mail: sumit.sxc13@gmail.com; Singh, S. N., E-mail: snsphyru@gmail.com; Prasad, K., E-mail: k.prasad65@gmail.com

    2016-05-06

    Lead-free solid solutions (1-x)Ba{sub 0.06}(Na{sub 1/2}Bi{sub 1/2}){sub 0.94}TiO{sub 3}-xNaNbO{sub 3} (0 ≤ x ≤ 1.0) were prepared by conventional ceramic fabrication technique. X-ray diffraction and Rietveld refinement analyses of these ceramics were carried out using X’Pert HighScore Plus software to determine the crystal symmetry, space group and unit cell dimensions. Rietveld refinement revealed that NaNbO{sub 3} with orthorhombic structure was completely diffused into Ba{sub 0.06}(Na{sub 1/2}Bi{sub 1/2}){sub 0.94}TiO{sub 3} lattice having the rhombohedral-tetragonal symmetry. EDS and SEM studies were carried out in order to evaluate the quality and purity of the compounds. SEM images showed a change in grain shapemore » with the increase of NaNbO{sub 3} content. FTIR spectra confirmed the formation of solid solution.« less

  3. Ce{sub 2}PdIn{sub 8}, Ce{sub 3}PdIn{sub 11} and Ce{sub 5}Pd{sub 2}In{sub 19}—members of homological series based on AuCu{sub 3}- and PtHg{sub 2}-type structural units

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

    Tursina, A.; Nesterenko, S.; Seropegin, Y.

    Crystal structures of three members of a unique homological series with the general formula Ce{sub m}Pd{sub n}In{sub 3m+2n} based on the AuCu{sub 3} and PtHg{sub 2} structure types were studied by single-crystal X-ray diffraction. The compounds crystallize with space group P4/mmm (Z=1) and the lattice parameters: a=4.6900(9) Å, c=12.185(6) Å for Ce{sub 2}PdIn{sub 8}, a=4.6846(8) Å, c=16.846(8) Å for Ce{sub 3}PdIn{sub 11}, and a=4.70120(10) Å, c=29.1359(4) Å for Ce{sub 5}Pd{sub 2}In{sub 19}. The crystal structures of Ce{sub 3}PdIn{sub 11} and Ce{sub 5}Pd{sub 2}In{sub 19} represent new types. The three structures constitute of [CeIn{sub 3}] cuboctahedra layers and [PdIn{sub 2}] rectangularmore » polyhedra layers, alternating along the tetragonal c-axis in accordance with the m:n proportion. The magnetic and electrical transport properties of the novel compounds Ce{sub 3}PdIn{sub 11} and Ce{sub 5}Pd{sub 2}In{sub 19} were investigated down to 1.72 K. Both indides are Curie–Weiss paramagnets due to the presence of fairly well localized 4f electrons of trivalent cerium ions. The electrical resistivity of both materials is dominated over an extended temperature range by strong spin–flip Kondo interactions with the characteristic temperature scale of 20–30 K. - Graphical abstract: TOC Figure Crystal structures of Ce{sub 3}PdIn{sub 11}, Ce{sub 2}PdIn{sub 8}, and Ce{sub 5}Pd{sub 2}In{sub 19}. Highlights: ► Large section of Ce–Pd–In phase diagram was examined. ► Three distinct ternary phases were identified, two of them for the first time. ► Crystal structures of two novel compounds constitute new structure types. ► The determined crystal structures show close mutual relationship. ► Ce{sub 3}PdIn{sub 11} and Ce{sub 5}Pd{sub 2}In{sub 19} are paramagnetic Kondo lattices.« less

  4. Synthesis and magnetic structure of the layered manganese oxide selenide Sr{sub 2}MnO{sub 2}Ag{sub 1.5}Se{sub 2}

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

    Blandy, Jack N.; Diamond Light Source Ltd., Harwell Science and Innovation Campus, Didcot OX11 0DE; Boskovic, Jelena C.

    The synthesis of a high-purity sample of the layered oxide selenide Sr{sub 2}MnO{sub 2}Ag{sub 1.5}Se{sub 2} is reported. At ambient temperature it crystallises in the space group I4/mmm with two formula units in the unit cell and lattice parameters a=4.08771(1) Å, c=19.13087(8) Å. The compound displays mixed-valent manganese in a formal oxidation state close to +2.5 and powder neutron diffraction measurements reveal that below the Néel temperature of 63(1) K this results in an antiferromagnetic structure which may be described as A-type, modelled in the magnetic space group P{sub I}4/mnc (128.410 in the Belov, Neronova and Smirnova (BNS) scheme) inmore » which localised Mn moments of 3.99(2) μ{sub B} are arranged in ferromagnetic layers which are coupled antiferromagnetically. In contrast to the isostructural compound Sr{sub 2}MnO{sub 2}Cu{sub 1.5}S{sub 2}, Sr{sub 2}MnO{sub 2}Ag{sub 1.5}Se{sub 2} does not display long range ordering of coinage metal ions and vacancies, nor may significant amounts of the coinage metal readily be deintercalated using soft chemical methods. - Graphical abstract: Sr{sub 2}MnO{sub 2}Ag{sub 1.5}Se{sub 2} containing mixed valent Mn ions undergoes magnetic ordering with ferromagnetic coupling within MnO{sub 2} sheets and antiferromagnetic coupling between MnO{sub 2} sheets. - Highlights: • High purity sample of Sr{sub 2}MnO{sub 2}Ag{sub 1.5}Se{sub 2} obtained. • Magnetic structure determined. • Compared with related mixed-valent manganite oxide chalcogenides.« less

  5. Crystallization and preliminary crystallographic studies of Hyp-1, a St John’s wort protein implicated in the biosynthesis of hypericin

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

    Fernandes, Humberto; Konieczna, Malgorzata; Kolodziejczyk, Robert

    2008-05-01

    The Hyp-1 protein suggested to be a phenolic oxidative-coupling enzyme involved in the biosynthesis of hypericin, a medicinally important natural compound found in St John’s wort (H. perforatum), has been expressed, purified and prepared in single-crystal form. According to a debated hypothesis, the biosynthesis from emodin of the medicinally important natural compound hypericin is catalyzed in St John’s wort (Hypericum perforatum) by the phenolic oxidative-coupling protein Hyp-1. Recombinant St John’s wort Hyp-1 has been overexpressed in Escherichia coli and obtained in single-crystal form. The crystals belong to the orthorhombic system, space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters amore » = 37.5, b = 76.7, c = 119.8 Å, contain two protein molecules in the asymmetric unit and diffract X-rays to 1.73 Å resolution.« less

  6. Syntheses and structural characterization of vanado-tellurites and vanadyl-selenites: SrVTeO{sub 5}(OH), Cd{sub 2}V{sub 2}Te{sub 2}O{sub 11}, Ca{sub 3}VSe{sub 4}O{sub 13}·H{sub 2}O and Ba{sub 2}VSe{sub 3}O{sub 10}

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

    Konatham, Satish; Vidyasagar, Kanamaluru, E-mail: kvsagar@iitm.ac.in

    Four new quaternary vanado-tellurites and vanadyl-selenites, namely, SrVTeO{sub 5}(OH)(1), Cd{sub 2}V{sub 2}Te{sub 2}O{sub 11}(2), Ca{sub 3}VSe{sub 4}O{sub 13}·H{sub 2}O(3) and Ba{sub 2}VSe{sub 3}O{sub 10}(4) have been synthesized and structurally characterized by single crystal X-ray diffraction. The oxidation state of vanadium is +5 in tellurites 1 and 2 and +4 in selenites 3 and 4. The structures of SrVTeO{sub 5}(OH)(1) and Cd{sub 2}V{sub 2}Te{sub 2}O{sub 11}(2) compounds consist of (VTeO{sub 5}(OH)){sup 2-} and (V{sub 2}Te{sub 2}O{sub 11}){sup 4-}anionic chains respectively, which are built from tetrahedral VO{sub 4} and disphenoidal TeO{sub 4} moieties. Similarly the structures of Ca{sub 3}VSe{sub 4}O{sub 13}·H{sub 2}O(3)more » and Ba{sub 2}VSe{sub 3}O{sub 10}(4) respectively contain (VSe{sub 2}O{sub 7}){sup 2-} and (VSe{sub 3}O{sub 10}){sup 4-} anionic chains, which are made up of octahedral VO{sub 6} and pyramidal SeO{sub 3} units. Compounds 1 and 3 have been characterized by thermogravimetric and infrared spectroscopic methods. Compounds 1 and 2 are wide band gap semiconductors. - Graphical abstract: Ca{sub 3}VSe{sub 4}O{sub 13}·H{sub 2}O and Ba{sub 2}VSe{sub 3}O{sub 10} compounds contain (VSe{sub 2}O{sub 7}){sup 2-} and (VSe{sub 3}O{sub 10}){sup 4-} chains. - Highlights: • Four new vanado-tellurites and vanadyl-selenites are synthesized. • Their structural features are different. • The vanado-tellurites are wide band gap semiconductors.« less

  7. Synthesis, crystal structure, and vibrational spectroscopic and UV-visible studies of Cs{sub 2}MnP{sub 2}O{sub 7}

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

    Kaoua, Saida; Krimi, Saida; Pechev, Stanislav

    2013-02-15

    A new member of the A{sub 2}MP{sub 2}O{sub 7} diphosphate family, Cs{sub 2}MnP{sub 2}O{sub 7}, has been synthesized and structurally characterized. The crystal structure was determined by single crystal X-Ray diffraction. Cs{sub 2}MnP{sub 2}O{sub 7} crystallizes in the orthorhombic system, space group Pnma ( Music-Sharp-Sign 62), with the unit cell parameters a=16.3398(3), b=5.3872(1), c=9.8872(2) A, Z=4 and V=870.33(3) A{sup 3}. The structure parameters were refined to a final R{sub 1}/wR{sub 2}=0.0194/0.0441 for 1650 observed reflections. The 2D framework of Cs{sub 2}MnP{sub 2}O{sub 7} structure consists of P{sub 2}O{sub 7} and MnO{sub 5} units. The corner-shared MnO{sub 5} and P{sub 2}O{submore » 7} units are alternately arranged along the b axis to form [(MnO)P{sub 2}O{sub 7}]{sub {infinity}} chains. These chains are interconnected by an oxygen atom to form sheets parallel to the (b, c) plane. The cesium atoms are located between the sheets in 9- and 10-fold coordinated sites. The infrared and Raman vibrational spectra have been investigated. A factor group analysis leads to the determination of internal modes of (P{sub 2}O{sub 7}) groups. UV-visible spectrum consists of weak bands, between 340 and 700 nm, assigned to the forbidden d-d transitions of Mn{sup 2+} ion, and of a strong band around 250 nm, attributed to the O--Mn charge transfer. - Graphical abstract: Structure of Cs{sub 2}MnP{sub 2}O{sub 7}: The 2D structure of Cs{sub 2}MnP{sub 2}O{sub 7} is built from P{sub 2}O{sub 7} diphosphate groups and MnO{sub 5} square pyramids which share corners and form [(MnO)P{sub 2}O{sub 7}]{sub {infinity}} chains along b axis. These chains are interconnected by an oxygen atom to form wavy (MnP{sub 2}O{sub 7}){sup 2-} sheets parallel to the (b, c) plane. The cesium ions are located between these sheets in the inter-layers space, in zigzag positions. Highlights: Black-Right-Pointing-Pointer A new diphosphate, Cs{sub 2}MnP{sub 2}O{sub 7}, has been synthesized and

  8. Cloning and characterization of an adenoviral vector for highly efficient and doxycycline – suppressible expression of bioactive human single – chain interleukin 12 in colon cancer

    PubMed Central

    Wulff, Holger; Krieger, Thorsten; Krüger, Karen; Stahmer, Ingrid; Thaiss, Friedrich; Schäfer, Hansjörg; Block, Andreas

    2007-01-01

    Background Interleukin-12 (IL-12) is well characterized to induce cellular antitumoral immunity by activation of NK-cells and T-lymphocytes. However, systemic administration of recombinant human IL-12 resulted in severe toxicity without perceptible therapeutic benefit. Even though intratumoral expression of IL-12 leads to tumor regression and long-term survival in a variety of animal models, clinical trials have not yet shown a significant therapeutic benefit. One major obstacle in the treatment with IL-12 is to overcome the relatively low expression of the therapeutic gene without compromising the safety of such an approach. Our objective was to generate an adenoviral vector system enabling the regulated expression of very high levels of bioactive, human IL-12. Results High gene expression was obtained utilizing the VP16 herpes simplex transactivator. Strong regulation of gene expression was realized by fusion of the VP16 to a tetracycline repressor with binding of the fusion protein to a flanking tetracycline operator and further enhanced by auto-regulated expression of its fusion gene within a bicistronic promoter construct. Infection of human colon cancer cells (HT29) at a multiplicity of infection (m.o.i.) of 10 resulted in the production of up to 8000 ng/106 cells in 48 h, thus exceeding any published vector system so far. Doxycycline concentrations as low as 30 ng/ml resulted in up to 5000-fold suppression, enabling significant reduction of gene expression in a possible clinical setting. Bioactivity of the human single-chain IL-12 was similar to purified human heterodimeric IL-12. Frozen sections of human colon cancer showed high expression of the coxsackie adenovirus receptor with significant production of human single chain IL-12 in colon cancer biopsies after infection with 3*107 p.f.u. Ad.3r-scIL12. Doxycycline mediated suppression of gene expression was up to 9000-fold in the infected colon cancer tissue. Conclusion VP16 transactivator-mediated and

  9. The A{sup 2+}Mn{sub 5}(SO{sub 4}){sub 6} family of triangular lattice, ferrimagnetic sulfates

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

    West, D.V., E-mail: barelytone@gmail.co; McQueen, T.M.; Posen, I.D.

    2009-06-15

    A new family of anhydrous sulfates, A{sup 2+}Mn{sub 5}(SO{sub 4}){sub 6} (A=Pb, Ba, Sr) is reported. The crystal structures of PbMn{sub 5}(SO{sub 4}){sub 6} and SrMn{sub 5}(SO{sub 4}){sub 6} are solved by powder X-ray and neutron diffraction. BaMn{sub 5}(SO{sub 4}){sub 6} is isostructural. PbMn{sub 5}(SO{sub 4}){sub 6} crystallizes with P3-bar symmetry and unit cell parameters of a=14.551(1) A and c=7.535(1) A. The structure has rich features, including dimers of face-sharing MnO{sub 6} octahedra, and two complementary triangular layers of Mn atoms. All compounds undergo a magnetic ordering transition at 10 K, below which, the magnetic susceptibility of the compounds variesmore » systematically with the radius of the non-magnetic cation. Low temperature neutron diffraction shows that the complementary triangular layers result in a ferrimagnet with a net moment corresponding to one high spin Mn{sup 2+} per unit cell, correlating well with the magnetization data. The non-magnetic variant PbMg{sub 5}(SO{sub 4}){sub 6} is also reported. - Graphical abstract: A new family sulfates, A{sup 2+}Mn{sub 5}(SO{sub 4}){sub 6} (A=Pb, Ba, Sr) is reported. Structures are solved by powder neutron diffraction. PbMn{sub 5}(SO{sub 4}){sub 6} is trigonal with lattice parameters of a=14.551(1) A and c=7.535(1) A. The structure has dimers of face-sharing MnO{sub 6} octahedra, and two complementary triangular layers of Mn atoms that result in a ferrimagnet. All compounds magnetically order at 10 K. Low field susceptibility varies systematically with non-magnetic cation radius.« less

  10. Intercalation of the layered solid acid HCa/sub 2/Nb/sub 3/O/sub 10/ by organic amines

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

    Jacobson, A.J.; Johnson, J.W.; Lewandowski, J.T.

    1987-01-01

    Layered compounds of formula C/sub n/H/sub 2n+1/NH/sub 3/Ca/sub 2/Nb/sub 3/O/sub 10/ are formed by reaction of n-alkylamines with the solid acid HCa/sub 2/Nb/sub 3/O/sub 10/. Other organic bases such as pyridine can also be intercalated. The lattice constants of the new compounds have been determined by powder X-ray diffraction. The unit cells are tetragonal with a axes that are unchanged with variation of the intercalated amine. The c axes lengths (layer spacings) vary systematically with the hydrocarbon chain length of the alkylamine.

  11. Antibodies specific for HT{sub m4}

    DOEpatents

    Lim, B.; Adra, C.N.; Lelias, J.M.

    1998-01-06

    The invention relates to a recombinant DNA molecule which encodes a HT{sub m4} protein, a transformed host cell which has been stably transfected with a DNA molecule which encodes a HT{sub m4} protein and a recombinant HT{sub m4} protein. The invention also relates to a method for detecting the presence of a hereditary atopy. 2 figs.

  12. Antibodies specific for HT.sub.m4

    DOEpatents

    Lim, Bing; Adra, Chaker N.; Lelias, Jean-Michel

    1998-01-01

    The invention relates to a recombinant DNA molecule which encodes a HT.sub.m4 protein, a transformed host cell which has been stably transfected with a DNA molecule which encodes a HT.sub.m4 protein and a recombinant HT.sub.m4 protein. The invention also relates to a method for detecting the presence of a hereditary atopy.

  13. Single crystals of (FeIn{sub 2}S{sub 4}){sub x} · (CuIn{sub 5}S{sub 8}){sub 1–x} alloys: Crystal structure, nuclear gamma resonance spectra, and thermal expansion

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

    Bodnar, I. V., E-mail: chemzav@bsuir.by; Zhafar, M. A.; Kasyuk, Yu. V.

    FeIn{sub 2}S{sub 4} and CuIn{sub 5}S{sub 8} compounds and (FeIn{sub 2}S{sub 4}){sub x} · (CuIn{sub 5}S{sub 8}){sub 1–x} alloy single crystals are grown by planar crystallization. It is shown that both of the initial FeIn{sub 2}S{sub 4} and CuIn{sub 5}S{sub 8} compounds and alloys on their basis crystallize with the formation of the cubic spinel structure. It is established that the unit-cell parameter a linearly varies with the composition parameter x. By means of nuclear gamma resonance spectroscopy in the transmission mode of measurements, the local states of iron ions in the alloys are studied. For the single crystals grownmore » in the study, thermal expansion is explored using the dilatometry technique, the thermal-expansion coefficients are determined, and the Debye temperature and rms (root-mean-square) dynamic displacements are calculated.« less

  14. Adenoviral-Mediated Imaging of Gene Transfer Using a Somatostatin Receptor-Cytosine Deaminase Fusion Protein

    PubMed Central

    Lears, Kimberly A.; Parry, Jesse J.; Andrews, Rebecca; Nguyen, Kim; Wadas, Thaddeus J.; Rogers, Buck E.

    2015-01-01

    Suicide gene therapy is a process by which cells are administered a gene that encodes a protein capable of converting a nontoxic prodrug into an active toxin. Cytosine deaminase (CD) has been widely investigated as a means of suicide gene therapy due to the enzyme’s ability to convert the prodrug 5-fluorocytosine (5-FC) into the toxic compound 5-fluorouracil (5-FU). However, the extent of gene transfer is a limiting factor in predicting therapeutic outcome. The ability to monitor gene transfer, non-invasively, would strengthen the efficiency of therapy. In this regard, we have constructed and evaluated a replication-deficient adenovirus (Ad) containing the human somatostatin receptor subtype 2 (SSTR2) fused with a C-terminal yeast CD gene for the non-invasive monitoring of gene transfer and therapy. The resulting Ad (AdSSTR2-yCD) was evaluated in vitro in breast cancer cells to determine the function of the fusion protein. These studies demonstrated that the both the SSTR2 and yCD were functional in binding assays, conversion assays, and cytotoxicity assays. In vivo studies similarly demonstrated the functionality using conversion assays, biodistribution studies, and small animal positron-emission tomography (PET) imaging studies. In conclusion, the fusion protein has been validated as useful for the non-invasive imaging of yCD expression and will be evaluated in the future for monitoring yCD-based therapy. PMID:25837665

  15. Crystal structure of Ca{sub 12}Al{sub 14}O{sub 32}Cl{sub 2} and luminescence properties of Ca{sub 12}Al{sub 14}O{sub 32}Cl{sub 2}:Eu{sup 2+}

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

    Iwata, Tomoyuki; Haniuda, Masahide; Fukuda, Koichiro

    2008-01-15

    The crystal structure of Ca{sub 12}Al{sub 14}O{sub 32}Cl{sub 2} was determined from laboratory X-ray powder diffraction data (CuK{alpha}{sub 1}) using the Rietveld method, with the anisotropic displacement parameters being assigned for all atoms. The crystal structure is cubic (space group I4-bar 3d, Z=2) with lattice dimensions a=1.200950(5) nm and V=1.73211(1) nm{sup 3}. The reliability indices calculated from the Rietveld method were R{sub wp}=8.48% (S=1.21), R{sub p}=6.05%, R{sub B}=1.27% and R{sub F}=1.01%. The validity of the structural model was verified by the three-dimensional electron density distribution, the structural bias of which was reduced as much as possible using the maximum-entropy methods-basedmore » pattern fitting (MPF). The reliability indices calculated from the MPF were R{sub B}=0.75% and R{sub F}=0.56%. In the structural model there are one Ca site, two Al sites, two O sites and one Cl site. This compound is isomorphous with Ca{sub 12}Al{sub 10.6}Si{sub 3.4}O{sub 32}Cl{sub 5.4}. Europium-doped sample Ca{sub 12}Al{sub 14}O{sub 32}Cl{sub 2}:Eu{sup 2+} was prepared and the photoluminescence properties were presented. The excitation spectrum consisted of two wide bands, which were located at about 268 and 324 nm. The emission spectrum, when excited at 324 nm, resulted in indigo light with a peak at about 442 nm. - Graphical abstract: A portion of the crystal structure of Ca{sub 12}Al{sub 14}O{sub 32}Cl{sub 2} showing eight-membered AlO{sub 4} rings and Ca-Cl-Ca unit.« less

  16. Super-resolution imaging and tracking of protein–protein interactions in sub-diffraction cellular space

    PubMed Central

    Liu, Zhen; Xing, Dong; Su, Qian Peter; Zhu, Yun; Zhang, Jiamei; Kong, Xinyu; Xue, Boxin; Wang, Sheng; Sun, Hao; Tao, Yile; Sun, Yujie

    2014-01-01

    Imaging the location and dynamics of individual interacting protein pairs is essential but often difficult because of the fluorescent background from other paired and non-paired molecules, particularly in the sub-diffraction cellular space. Here we develop a new method combining bimolecular fluorescence complementation and photoactivated localization microscopy for super-resolution imaging and single-molecule tracking of specific protein–protein interactions. The method is used to study the interaction of two abundant proteins, MreB and EF-Tu, in Escherichia coli cells. The super-resolution imaging shows interesting distribution and domain sizes of interacting MreB–EF-Tu pairs as a subpopulation of total EF-Tu. The single-molecule tracking of MreB, EF-Tu and MreB–EF-Tu pairs reveals intriguing localization-dependent heterogonous dynamics and provides valuable insights to understanding the roles of MreB–EF-Tu interactions. PMID:25030837

  17. Sub-diffusion and trapped dynamics of neutral and charged probes in DNA-protein coacervates

    NASA Astrophysics Data System (ADS)

    Arfin, Najmul; Yadav, Avinash Chand; Bohidar, H. B.

    2013-11-01

    The physical mechanism leading to the formation of large intermolecular DNA-protein complexes has been studied. Our study aims to explain the occurrence of fast coacervation dynamics at the charge neutralization point, followed by the appearance of smaller complexes and slower coacervation dynamics as the complex experiences overcharging. Furthermore, the electrostatic potential and probe mobility was investigated to mimic the transport of DNA / DNA-protein complex in a DNA-protein complex coacervate medium [N. Arfin and H. B. Bohidar, J. Phys. Chem. B 116, 13192 (2012)] by assigning neutral, negative, or positive charge to the probe particle. The mobility of the neutral probe was maximal at low matrix concentrations and showed random walk behavior, while its mobility ceased at the jamming concentration of c = 0.6, showing sub-diffusion and trapped dynamics. The positively charged probe showed sub-diffusive random walk followed by trapped dynamics, while the negatively charged probe showed trapping with occasional hopping dynamics at much lower concentrations. Sub-diffusion of the probe was observed in all cases under consideration, where the electrostatic interaction was used exclusively as the dominant force involved in the dynamics. For neutral and positive probes, the mean square displacement ⟨R2⟩ exhibits a scaling with time as ⟨R2⟩ ˜ tα, distinguishing random walk and trapped dynamics at α = 0.64 ± 0.04 at c = 0.12 and c = 0.6, respectively. In addition, the same scaling factors with the exponent β = 0.64 ± 0.04 can be used to distinguish random walk and trapped dynamics for the neutral and positive probes using the relation between the number of distinct sites visited by the probe, S(t), which follows the scaling, S(t) ˜ tβ/ln (t). Our results established the occurrence of a hierarchy of diffusion dynamics experienced by a probe in a dense medium that is either charged or neutral.

  18. Crystallization and initial X-ray diffraction studies of scaffolding protein (gp7) of bacteriophage ϕ29

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

    Badasso, Mohammed O., E-mail: badas001@umn.edu; Anderson, Dwight L.; Department of Oral Science, University of Minnesota, Minneapolis, MN 55455

    2005-04-01

    ϕ29 bacteriophage scaffolding protein (gp7) has been overproduced in E. coli, purified, crystallized and characterized by X-ray diffraction. Two distinct crystal forms were obtained and a diffraction data set was collected to 1.8 Å resolution. The Bacillus subtilis bacteriophage ϕ29 scaffolding protein (gp7) has been crystallized by the hanging-drop vapour-diffusion method at 293 K. Two new distinct crystal forms that both differed from a previously crystallized and solved scaffolding protein were grown under the same conditions. Form I belongs to the primitive tetragonal space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 77.13, c = 37.12 Å.more » Form II crystals exhibit an orthorhombic crystal form, with space group C222 and unit-cell parameters a = 107.50, b = 107. 80, c = 37.34 Å. Complete data sets have been collected to 1.78 and 1.80 Å for forms I and II, respectively, at 100 K using Cu Kα X-rays from a rotating-anode generator. Calculation of a V{sub M} value of 2.46 Å{sup 3} Da{sup −1} for form I suggests the presence of one molecule in the asymmetric unit, corresponding to a solvent content of 50.90%, whereas form II has a V{sub M} of 4.80 Å{sup 3} Da{sup −1} with a solvent content of 48.76% and two molecules in the asymmetric unit. The structures of both crystal forms are being determined by the molecular-replacement method using the coordinates of the published crystal structure of gp7.« less

  19. Live-cell topology assessment of URG7, MRP6{sub 102} and SP-C using glycosylatable green fluorescent protein in mammalian cells

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

    Lee, Hunsang; Lara, Patricia; Ostuni, Angela

    2014-08-08

    Highlights: • Glycosylatable GFP (gGFP) is developed for the use in mammalian cells. • gGFP selectively loses its fluorescence upon N-linked glycosylation in the ER lumen. • Differential fluorescence/glycosylation pattern probes membrane protein topology. • Membrane topology of URG7, MRP6{sub 102}, and SP-C was determined by gGFP tagging in vivo. - Abstract: Experimental tools to determine membrane topology of a protein are rather limited in higher eukaryotic organisms. Here, we report the use of glycosylatable GFP (gGFP) as a sensitive and versatile membrane topology reporter in mammalian cells. gGFP selectively loses its fluorescence upon N-linked glycosylation in the ER lumen.more » Thus, positive fluorescence signal assigns location of gGFP to the cytosol whereas no fluorescence signal and a glycosylated status of gGFP map the location of gGFP to the ER lumen. By using mammalian gGFP, the membrane topology of disease-associated membrane proteins, URG7, MRP6{sub 102}, SP-C(Val) and SP-C(Leu) was confirmed. URG7 is partially targeted to the ER, and inserted in C{sub in} form. MRP6{sub 102} and SP-C(Leu/Val) are inserted into the membrane in C{sub out} form. A minor population of untargeted SP-C is removed by proteasome dependent quality control system.« less

  20. Synthesis and structure of R{sub 2}[UO{sub 2}(NO{sub 3}){sub 2}(NCS){sub 2}] (R = Rb or Cs)

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

    Serezhkin, V. N., E-mail: lserezh@samsu.ru; Peresypkina, E. V.; Grigor’eva, V. A.

    2015-01-15

    Crystals Rb{sub 2}[UO{sub 2}(NO{sub 3}){sub 2}(NCS){sub 2}] (I) and Cs{sub 2}[UO{sub 2}(NO{sub 3}){sub 2}(NCS){sub 2}] (II) have been synthesized and studied by IR spectroscopy and X-ray diffraction. Crystals I are monoclinic, with the following parameters: a = 12.2118(5) Å, b = 10.2545(3) Å, c = 11.8754(4) Å, β = 110.287(1)°, sp. gr. C2/c, Z = 4, and R = 0.0523. Crystals II are orthorhombic, with a = 13.7309(3) Å, b = 10.5749(2) Å, c = 10.1891(2) Å, sp. gr. Pnma, Z = 4, and R = 0.0411. The basic structural units of crystals I and II are one-core complexes [UO{submore » 2}(NO{sub 3}){sub 2}(NCS){sub 2}]{sup 2−}, which belong to the crystallochemical group cis-AB{sub 2}{sup 01}M{sub 2}{sup 1} (A = UO{sub 2}{sup 2+}, B{sup 01} = NO{sub 3}{sup −}, M{sup 1} = NCS{sup −}), which are combined into a framework via electrostatic interactions with ions of alkaline metals R (R = Rb or Cs). The structural features of crystals I and II, which condition the formation of [UO{sub 2}(NO{sub 3}){sub 2}(NCS){sub 2}]{sup 2−} complexes with a cis rather than a trans position of isothiocyanate ions in the coordination sphere of uranyl ions, are discussed.« less

  1. Magnetochemistry of the tetrahaloferrate (III) ions. 7. Crystal structure and magnetic ordering in (pyridinium){sub 3}Fe{sub 2}Br{sub 9}

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

    Lowe, C.B.; Shaviv, R.; Carlin, R.L.

    1994-07-06

    A monoclinic crystal structure was found by X-ray diffraction for bis [pyridinium tetrabromferrate(III)]-pyridinium bromide. The double salt contains two slightly distorted [FeBr{sub 4}]{sup -} tetrahedra, three pyridinium rings, and an uncoordinated halide in each asymmetric unit, as is characteristic of the A{sub 3}Fe{sub 2}X{sub 9} series of compounds. Unit cell parameters, monoclinic space group P2{sub 1}, are a = 7.656(3) {angstrom}, b = 14.237(5) {angstrom}, c = 13.725(5) {angstrom}, {beta} = 93.42(3){degrees}, and V = 1493(1) {angstrom}{sup 3}, using Mo K{alpha} radiation {lambda} = 0.710 69 {angstrom}, {rho}{sub calc} = 2.38 g cm{sup -3}, and Z = 2. The tetrahedramore » are aligned with their 3-fold axes parallel to the crystallographic c axis. Bond lengths (Fe-Br) range from 2.271(9) {angstrom} to 2.379(9) {angstrom} for the two different slightly distorted tetrahedral units. Magnetic susceptibility studies show that the material orders three-dimensionally at 7.4 {+-} 0.2 K. The data are compared to a HTS expansion of 1/{sub {chi}} for the S = 5/2 three-dimensional Heisenberg model antiferromagnet for a sc lattice with g = 1.98 and J/k{sub B} = -0.43 K. The specific heat measurements indicate two odd-shaped {lambda} features, at 7.3 and 8 K.« less

  2. Method for generating O.sub.2-rich gas from air using water

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

    Nakano, Anna; Nakano, Jinichiro; Bennett, James P.

    The present disclosure is directed to a method for enriching an inlet air stream utilizing a number of enrichment sub-units connected in series, where each enrichment sub-unit conducts both a dissolution and degasification cycle. Each enrichment sub-unit comprises a compressor, an aeration unit, a deaeration unit, and a pump for the recirculation of water between the aeration and deaeration units. The methodology provides a manner in which the relationship between the respective Henry's coefficients of the oxygen and nitrogen in water may be exploited to enrich the O.sub.2 volume percent and diminish the N.sub.2 volume percent over repeated dissolution andmore » degasification cycles. By utilizing a number of enrichment sub-units connected in series, the water contained in each enrichment sub-unit acts to progressively increase the O.sub.2 volume percent. Additional enrichment sub-units may be added and utilized until the O.sub.2 volume percent equals or exceeds a target O.sub.2 volume percent. In a particular embodiment, air having a general composition of about 78 vol. % N.sub.2 and 21 vol. % O.sub.2 is progressively enriched to provide a final mixture of about 92% vol. % O.sub.2 and 8% vol. % N.sub.2.« less

  3. Process for protein enrichment of cassava by solid substrate fermentation in rural conditions

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

    Daubresse, P.; Ntibashirwa, S.; Gheysen, A.

    1987-06-01

    An artisanal static process for protein enrichment of cassava by solid-state fermentation, developed in laboratory and tested on pilot units in Burundi (Central Africa), provides enriched cassava containing 10.7% of dry matter protein versus 1% before fermentation. Cassava chips, processed into granules of 2-4-mm diameter, are moistened (40% water content) and steamed. After cooling to 40 degrees C, cassava is mixed with a nutritive solution containing the inoculum (Rhizopus oryzae, strain MUCL 28627) and providing the following per 100 g dry matter: 3.4 g urea, 1.5 g KH/sub 2/PO/sub 4/, O.8 g MgSO/sub 4/.7H/sub 2/O, and 22.7 g citric acid.more » For the fermentation, cassava, with circa 60% moisture content, is spread in a thin layer (2-3 cm thick) on perforated trays and slid into an aerated humidified enclosure. The incubation lasts more or less 65 hours. The production of protein enriched cassava is 3.26 kg dry matter/square m tray. The effects of the variation of the nutritive solution composition and the inoculum conservation period on the protein production are equally discussed. (Refs. 37).« less

  4. Pancreatic Transduction by Helper-Dependent Adenoviral Vectors via Intraductal Delivery

    PubMed Central

    Morró, Meritxell; Teichenne, Joan; Jimenez, Veronica; Kratzer, Ramona; Marletta, Serena; Maggioni, Luca; Mallol, Cristina; Ruberte, Jesus; Kochanek, Stefan; Bosch, Fatima

    2014-01-01

    Abstract Pancreatic gene transfer could be useful to treat several diseases, such as diabetes mellitus, cystic fibrosis, chronic pancreatitis, or pancreatic cancer. Helper-dependent adenoviral vectors (HDAds) are promising tools for gene therapy because of their large cloning capacity, high levels of transgene expression, and long-term persistence in immunocompetent animals. Nevertheless, the ability of HDAds to transduce the pancreas in vivo has not been investigated yet. Here, we have generated HDAds carrying pancreas-specific expression cassettes, that is, driven either by the elastase or insulin promoter, using a novel and convenient plasmid family and homologous recombination in bacteria. These HDAds were delivered to the pancreas of immunocompetent mice via intrapancreatic duct injection. HDAds, encoding a CMV-GFP reporter cassette, were able to transduce acinar and islet cells, but transgene expression was lost 15 days postinjection in correlation with severe lymphocytic infiltration. When HDAds encoding GFP under the control of the specific elastase promoter were used, expression was detected in acinar cells, but similarly, the expression almost disappeared 30 days postinjection and lymphocytic infiltration was also observed. In contrast, long-term transgene expression (>8 months) was achieved with HDAds carrying the insulin promoter and the secretable alkaline phosphatase as the reporter gene. Notably, transduction of the liver, the preferred target for adenovirus, was minimal by this route of delivery. These data indicate that HDAds could be used for pancreatic gene therapy but that selection of the expression cassette is of critical importance to achieve long-term expression of the transgene in this tissue. PMID:25046147

  5. Combination recombinant simian or chimpanzee adenoviral vectors for vaccine development.

    PubMed

    Cheng, Cheng; Wang, Lingshu; Ko, Sung-Youl; Kong, Wing-Pui; Schmidt, Stephen D; Gall, Jason G D; Colloca, Stefano; Seder, Robert A; Mascola, John R; Nabel, Gary J

    2015-12-16

    Recombinant adenoviral vector (rAd)-based vaccines are currently being developed for several infectious diseases and cancer therapy, but pre-existing seroprevalence to such vectors may prevent their use in broad human populations. In this study, we investigated the potential of low seroprevalence non-human primate rAd vectors to stimulate cellular and humoral responses using HIV/SIV Env glycoprotein (gp) as the representative antigen. Mice were immunized with novel simian or chimpanzee rAd (rSAV or rChAd) vectors encoding HIV gp or SIV gp by single immunization or in heterologous prime/boost combinations (DNA/rAd; rAd/rAd; rAd/NYVAC or rAd/rLCM), and adaptive immunity was assessed. Among the rSAV and rChAd tested, rSAV16 or rChAd3 vector alone generated the most potent immune responses. The DNA/rSAV regimen also generated immune responses similar to the DNA/rAd5 regimen. rChAd63/rChAd3 and rChAd3 /NYVAC induced similar or even higher levels of CD4+ and CD8+ T-cell and IgG responses as compared to rAd28/rAd5, one of the most potent combinations of human rAds. The optimized vaccine regimen stimulated improved cellular immune responses and neutralizing antibodies against HIV compared to the DNA/rAd5 regimen. Based on these results, this type of novel rAd vector and its prime/boost combination regimens represent promising candidates for vaccine development. Published by Elsevier Ltd.

  6. Communication: A simple full range analytical potential for H{sub 2} b{sup 3}∑{sub u}{sup +}, H–He {sup 2}∑{sup +}, and He{sub 2} {sup 1}∑{sub g}{sup +}

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

    Warnecke, Sascha; Toennies, J. Peter, E-mail: jtoenni@gwdg.de; Tang, K. T.

    The Tang-Toennies potential for the weakly interacting systems H{sub 2} b{sup 3}Σ{sub u}{sup +}, H–He {sup 2}Σ{sup +}, and He{sub 2} {sup 1}Σ{sub g}{sup +} is extended down to the united atom limit of vanishing internuclear distance. A simple analytic expression connects the united atom limiting potential with the Tang-Toennies potential in the well region. The new potential model is compared with the most recent ab initio calculations for all three systems. The agreement is better than 20% (H{sub 2} and He{sub 2}) or comparable with the differences in the available ab initio calculations (H–He) over six orders of magnitudemore » corresponding to the entire range of internuclear distances.« less

  7. Serine 192 in the tiny RS repeat of the adenoviral L4-33K splicing enhancer protein is essential for function and reorganization of the protein to the periphery of viral replication centers

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

    Oestberg, Sara, E-mail: sara.ostberg@imbim.uu.se; Toermaenen Persson, Heidi, E-mail: heidi.tormanen.persson@imbim.uu.se; Akusjaervi, Goeran, E-mail: goran.akusjarvi@imbim.uu.se

    2012-11-25

    The adenovirus L4-33K protein is a key regulator involved in the temporal shift from early to late pattern of mRNA expression from the adenovirus major late transcription unit. L4-33K is a virus-encoded alternative splicing factor, which enhances processing of 3 Prime splice sites with a weak sequence context. Here we show that L4-33K expressed from a plasmid is localized at the nuclear margin of uninfected cells. During an infection L4-33K is relocalized to the periphery of E2A-72K containing viral replication centers. We also show that serine 192 in the tiny RS repeat of the conserved carboxy-terminus of L4-33K, which ismore » critical for the splicing enhancer function of L4-33K, is necessary for the nuclear localization and redistribution of the protein to viral replication sites. Collectively, our results show a good correlation between the activity of L4-33K as a splicing enhancer protein and its localization to the periphery of viral replication centers.« less

  8. Crystal structure, oxidation state and magnetism of Sr{sub x}La{sub 2−x}Cu{sub 0.5}Ru{sub 0.5}O{sub 4} (x=1, 1.5)

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

    Lü, Minfeng, E-mail: m.f.lv@ciac.jl.cn; Deng, Xiaolong; Waerenborgh, João C.

    2014-03-15

    Sr{sub x}La{sub 2−x}Cu{sub 0.5}Ru{sub 0.5}O{sub 4} (x=1, 1.5) oxides with K{sub 2}NiF{sub 4}-type structure were prepared by solid state reaction and characterized by powder X-ray diffraction, X-ray photoelectron spectroscopy, magnetic and electrical resistivity measurements. The SrLaCu{sub 0.5}Ru{sub 0.5}O{sub 4} phase was obtained for the first time with a negligible amount of impurities. The octahedral Cu/RuO{sub 6} units are more elongated in SrLaCu{sub 0.5}Ru{sub 0.5}O{sub 4} than in Sr{sub 1.5}La{sub 0.5}Cu{sub 0.5}Ru{sub 0.5}O{sub 4} indicating a greater extent of static Jahn–Teller distortion. XPS suggests that mixed ion pairs Ru{sup 5+}/Ru{sup 4+}↔Cu{sup +}/Cu{sup 2+} are present in SrLaCu{sub 0.5}Ru{sub 0.5}O{sub 4}, whilemore » Ru remains as Ru{sup 5+} and Cu as Cu{sup 2+} in Sr{sub 1.5}La{sub 0.5}Cu{sub 0.5}Ru{sub 0.5}O{sub 4}. Both samples show spin-glass behavior, which can be explained by competition between ferromagnetic and antiferromagnetic superexchange interactions. The negative Weiss temperature estimated for SrLaCu{sub 0.5}Ru{sub 0.5}O{sub 4}, −318 K, is significantly lower than −11.5 K deduced for Sr{sub 1.5}La{sub 0.5}Cu{sub 0.5}Ru{sub 0.5}O{sub 4} which may be related to the higher static Jahn–Teller distortion in the former oxide. -- Graphical abstract: SrLaCu{sub 0.5}Ru{sub 0.5}O{sub 4} with K{sub 2}NiF{sub 4}-type structure show a larger static Jahn–Teller distortion than Sr{sub 1.5}La{sub 0.5}Cu{sub 0.5}Ru{sub 0.5}O{sub 4}, which may be related to stronger antiferromagnetic superexchange interactions. Highlights: • SrLaCu{sub 0.5}Ru{sub 0.5}O{sub 4} (I) larger Jahn–Teller (J–T) distortion than Sr{sub 1.5}La{sub 0.5}Cu{sub 0.5}Ru{sub 0.5}O{sub 4} (II). • Octahedral Cu/RuO{sub 6} units are more elongated in I than in II. • Mixed ion pairs Ru{sup 5+}/Ru{sup 4+}↔Cu{sup +}/Cu{sup 2+} are present in I, while Ru remains as Ru{sup 5+} and Cu as Cu{sup 2+} in II. • Negative Weiss temperature of I significantly lower

  9. Li{sub 2}Ca{sub 2}Si{sub 2}O{sub 7}: Structural, spectroscopic and computational studies on a sorosilicate

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

    Kahlenberg, Volker, E-mail: volker.kahlenberg@uibk.ac.at; Brunello, Emanuele; Hejny, Clivia

    2015-05-15

    Synthesis experiments in the system Li{sub 2}O–CaO–SiO{sub 2} resulted in the formation of single-crystals of Li{sub 2}Ca{sub 2}Si{sub 2}O{sub 7}. Structural investigations were based on single-crystal diffraction. At ambient conditions the compound has the following basic crystallographic data: hexagonal symmetry, space group P6{sub 1}22, a=5.0961(2) Å, c=41.264(2) Å, V=928.07(6) Å{sup 3}, Z=6. Structure solution was performed using direct methods. The final least-squares refinement calculations converged at a residual of R(|F|)=0.0260. From a structural point the lithium calcium silicate belongs to the group of pyrosilicates containing [Si{sub 2}O{sub 7}]-groups. Additional lithium and calcium cations are incorporated between the silicate dimers andmore » are coordinated by four and six nearest oxygen neighbours, respectively. Each [LiO{sub 4}]-tetrahedron shares two common corners with directly neighboring tetrahedra forming zweier single-chains which are running parallel to 〈1 0 0〉 in z-levels defined by the presence of the 6{sub 1}{sup [0} {sup 0} {sup 1]}-screw axes. From the corner-sharing [LiO{sub 4}]- and [SiO{sub 4}]-moieties a three dimensional framework can be constructed. An interesting feature of this framework is the presence of an O{sup [3]}-type bridging oxygen linking three tetrahedra (one [LiO{sub 4}]- and two [SiO{sub 4}]-units). Structural similarities with other silicates are discussed in detail. The high-temperature behavior of the Si–O, Ca–O and Li–O bond distances in Li{sub 2}Ca{sub 2}Si{sub 2}O{sub 7} was investigated by in{sub -}situ single-crystal X-ray diffraction in the range between 65 and 700 °C. From the evolution of the lattice parameters, the thermal expansion tensor α{sub ij} has been determined. The structural characterization has been supplemented by micro-Raman spectroscopy. Interpretation of the spectroscopic data including the allocation of the bands to certain vibrational species has been aided by DFT

  10. Zea mays Taxilin Protein Negatively Regulates Opaque-2 Transcriptional Activity by Causing a Change in Its Sub-Cellular Distribution

    PubMed Central

    Zhang, Nan; Qiao, Zhenyi; Liang, Zheng; Mei, Bing; Xu, Zhengkai; Song, Rentao

    2012-01-01

    Zea mays (maize) Opaque-2 (ZmO2) protein is an important bZIP transcription factor that regulates the expression of major storage proteins (22-kD zeins) and other important genes during maize seed development. ZmO2 is subject to functional regulation through protein-protein interactions. To unveil the potential regulatory network associated with ZmO2, a protein-protein interaction study was carried out using the truncated version of ZmO2 (O2-2) as bait in a yeast two-hybrid screen with a maize seed cDNA library. A protein with homology to Taxilin was found to have stable interaction with ZmO2 in yeast and was designated as ZmTaxilin. Sequence analysis indicated that ZmTaxilin has a long coiled-coil domain containing three conserved zipper motifs. Each of the three zipper motifs is individually able to interact with ZmO2 in yeast. A GST pull-down assay demonstrated the interaction between GST-fused ZmTaxilin and ZmO2 extracted from developing maize seeds. Using onion epidermal cells as in vivo assay system, we found that ZmTaxilin could change the sub-cellular distribution of ZmO2. We also demonstrated that this change significantly repressed the transcriptional activity of ZmO2 on the 22-kD zein promoter. Our study suggests that a Taxilin-mediated change in sub-cellular distribution of ZmO2 may have important functional consequences for ZmO2 activity. PMID:22937104

  11. [Effects of adenoviral vector containing human angiotensin II type 1 receptor antisense cDNA on biological action of human pulmonary artery smooth muscle cells].

    PubMed

    Tu, Ming-li; Wang, Han-qin; Lei, Huai-ding; Luo, Guo-shi; Liu, Xian-jun; Liu, Wei-shun; Xiong, Chang; Liu, Yu-quan; Ren, Si-qun

    2005-04-01

    To investigate the effect of human angiotensin II (AngII) type 1 receptor (AT(1)R) antisense cDNA (ahAT(1)) on migration, proliferation, and apoptosis of cultured human pulmonary artery smooth muscle cells (PASMC). Two recombinant adenoviral vectors, AdCMVahAT(1) containing full length antisense cDNA targeting to human AT(1)R mRNA, and AdCMVLacZ containing LacZ, were constructed by orientation clone technology and homologous recombination. The PASMC was divided into 3 groups (DMEM, AdCMVLacZ, AdCMVahAT(1)) and different interventions were given to different groups. AT(1)R expression was detected by RT-PCR and immunohistochemistry method; migration of PASMC was measured by Boyden's Chamer method. Other PASMC was divided into 4 groups (DMEM, AngII, AdCMVLacZ + AngII and AdCMVahAT(1) + AngII), and only the last 2 groups were respectively transfected with AdCMVLacZ and AdCMVahAT(1) before administration of AngII. From 6 h to 96 h after stimulation by AngII (10(-7) mol/L), proliferation index (PI) and apoptosis of PASMC were determined by flow cytometry. At the 48 h the level of AT(1)R mRNA was significantly less in PASMC transfected AdCMVahAT(1) than that in group DMEM and in group AdCMVLacZ. The protein level showed a same difference (P < 0.01). At 24 h the migration distance of PASMC also was significantly less in group AdCMVahAT(1) than that in group DMEM and Group AdCMVLacZ (P < 0.01). Stimulated by AngII for 48 h, in group AngII the PI of PASMC markedly increased (P < 0.01 vs group DMEM). But in Group AdCMVahAT(1) + AngII PI of PASMC clearly decreased (P < 0.01 vs group AngII and group DMEM respectively). There was no statistic difference of PI between group AdCMVLacZ + AngII and group AngII. Moreover, apoptosis peak emerged only in group AdCMVahAT(1) + AngII. The rate of apoptosis in those PASMC used AdCMVahAT(1) and AngII was 24.70 +/- 4.04 (P < 0.01 vs the other 3 groups respectively). These results indicate that AngII stimulates proliferation via AT(1

  12. Crystal structure of the mineral (Na,Ca,K){sub 2}(Ca,Na){sub 4}(Mg,Fe){sub 5}(Mg,Fe,Ti){sub 5}[Si{sub 12}Al{sub 4}O{sub 44}](F,O){sub 4}: a triclinic representative of the amphibole family

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

    Rastsvetaeva, R. K., E-mail: rast@ns.crys.ras.ru; Aksenov, S. M.

    2012-05-15

    A mineral belonging to the amphibole family found at the Rothenberg paleovolcano (Eifel, Germany) was studied by single-crystal X-ray diffraction. The triclinic pseudomonoclinic unit-cell parameters are a = 5.3113(1) Angstrom-Sign , b = 18.0457(3) Angstrom-Sign ; c = 9.8684(2) Angstrom-Sign , {alpha} = 90.016(2) Degree-Sign , {beta} = 105.543(4) Degree-Sign , {gamma} = 89.985(2) Degree-Sign . The structure was solved by direct methods in sp. gr. P1 and refined to the R factor of 2.7% based on 6432 reflections with |F| > 3{sigma}(F) taking into account twinning. The mineral with the idealized formula (Na,Ca,K){sub 2}(Ca,Na){sub 4}(Mg,Fe){sub 5}(Mg,Fe,Ti){sub 5}[Si{sub 12}Al{sub 4}O{submore » 44}](F,O){sub 4} has some symmetry and structural features that distinguish it from other minerals of this family.« less

  13. Interface induced high temperature superconductivity in single unit-cell FeSe on SrTiO{sub 3}(110)

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

    Zhou, Guanyu; Zhang, Ding; Liu, Chong

    2016-05-16

    We report high temperature superconductivity in one unit-cell (1-UC) FeSe films grown on SrTiO{sub 3} (STO)(110) substrate by molecular beam epitaxy. By in-situ scanning tunneling microscopy measurement, we observe a superconducting gap as large as 17 meV on the 1-UC FeSe films. Transport measurements on 1-UC FeSe/STO(110) capped with FeTe layers reveal superconductivity with an onset transition temperature (T{sub C}) of 31.6 K and an upper critical magnetic field of 30.2 T. We also find that T{sub C} can be further increased by external electric field although the effect is weaker than that on STO(001) substrate.

  14. {[Ga(en){sub 3}]{sub 2}(Ge{sub 2}Te{sub 15})}n : a polymeric semiconducting polytelluride with boat-shaped Te{sub 8}{sup 4-} rings and cross-shaped Te{sub 5}{sup 6-} units.

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

    Zhang, Q.; Malliakas, C. D.; Kanatzidis, M. G.

    2009-11-11

    The reaction of the Zintl compound K{sub 4}Ge{sub 9} with Te and Ga{sub 2}Te{sub 3} in ethylenediamine (en) at 190 C gave the germanium polytelluride {l_brace}[Ga(en){sub 3}]{sub 2}[(GeTe){sub 2}(Te{sub 5}){sup 6-}(Te{sub 8}){sup 4-}]{r_brace}n (1). The single-crystal structure analysis revealed that 1 has two different polytelluride fragments: cross-shaped 36-e- TeTe{sub 4}{sup 6-} anions and boat-shaped 52-e- Te{sup 8}{sup 4-} rings. The new material is a p-type semiconductor at room temperature and switches to n-type at 380 K.

  15. Human Adenovirus Core Protein V Is Targeted by the Host SUMOylation Machinery To Limit Essential Viral Functions.

    PubMed

    Freudenberger, Nora; Meyer, Tina; Groitl, Peter; Dobner, Thomas; Schreiner, Sabrina

    2018-02-15

    Human adenoviruses (HAdV) are nonenveloped viruses containing a linear, double-stranded DNA genome surrounded by an icosahedral capsid. To allow proper viral replication, the genome is imported through the nuclear pore complex associated with viral core proteins. Until now, the role of these incoming virion proteins during the early phase of infection was poorly understood. The core protein V is speculated to bridge the core and the surrounding capsid. It binds the genome in a sequence-independent manner and localizes in the nucleus of infected cells, accumulating at nucleoli. Here, we show that protein V contains conserved SUMO conjugation motifs (SCMs). Mutation of these consensus motifs resulted in reduced SUMOylation of the protein; thus, protein V represents a novel target of the host SUMOylation machinery. To understand the role of protein V SUMO posttranslational modification during productive HAdV infection, we generated a replication-competent HAdV with SCM mutations within the protein V coding sequence. Phenotypic analyses revealed that these SCM mutations are beneficial for adenoviral replication. Blocking protein V SUMOylation at specific sites shifts the onset of viral DNA replication to earlier time points during infection and promotes viral gene expression. Simultaneously, the altered kinetics within the viral life cycle are accompanied by more efficient proteasomal degradation of host determinants and increased virus progeny production than that observed during wild-type infection. Taken together, our studies show that protein V SUMOylation reduces virus growth; hence, protein V SUMOylation represents an important novel aspect of the host antiviral strategy to limit virus replication and thereby points to potential intervention strategies. IMPORTANCE Many decades of research have revealed that HAdV structural proteins promote viral entry and mainly physical stability of the viral genome in the capsid. Our work over the last years showed that this

  16. Identifying essential proteins based on sub-network partition and prioritization by integrating subcellular localization information.

    PubMed

    Li, Min; Li, Wenkai; Wu, Fang-Xiang; Pan, Yi; Wang, Jianxin

    2018-06-14

    Essential proteins are important participants in various life activities and play a vital role in the survival and reproduction of living organisms. Identification of essential proteins from protein-protein interaction (PPI) networks has great significance to facilitate the study of human complex diseases, the design of drugs and the development of bioinformatics and computational science. Studies have shown that highly connected proteins in a PPI network tend to be essential. A series of computational methods have been proposed to identify essential proteins by analyzing topological structures of PPI networks. However, the high noise in the PPI data can degrade the accuracy of essential protein prediction. Moreover, proteins must be located in the appropriate subcellular localization to perform their functions, and only when the proteins are located in the same subcellular localization, it is possible that they can interact with each other. In this paper, we propose a new network-based essential protein discovery method based on sub-network partition and prioritization by integrating subcellular localization information, named SPP. The proposed method SPP was tested on two different yeast PPI networks obtained from DIP database and BioGRID database. The experimental results show that SPP can effectively reduce the effect of false positives in PPI networks and predict essential proteins more accurately compared with other existing computational methods DC, BC, CC, SC, EC, IC, NC. Copyright © 2018 Elsevier Ltd. All rights reserved.

  17. [The virological and epidemiological aspects of human adenoviral conjunctivitis in Tunisia].

    PubMed

    Fedaoui, N; Ben Ayed, N; Ben Yahia, A; Matri, L; Nacef, L; Triki, H

    2017-01-01

    Human adenoviruses (HAdV) are the main cause of viral conjunctivitis. In Tunisia and North Africa more generally, there is no regular nationwide surveillance program that monitors viruses causing conjunctivitis and keratoconjunctivitis. In this study, we report the results of HAdV screening in conjunctival samples collected for over 14 years in Tunisia. A total of 282 conjunctival samples received between 2000 and 2013 were investigated. Detection and identification of genotype were performed by PCR-sequencing at the hexon gene; 64.5% of samples (n=182) revealed positive by PCR detection without correlation noted between infection, age, sex, social class or clinical manifestations of viral conjunctivitis. HAdV-D8 was the largely predominant genotype in Tunisia, representing 81.3% of all isolates, and was detected continuously from 2000 to 2013. Minor co-circulating genotypes were also identified - HAdV-E4, HAdV-B3, B55 and HAdV-B7 - accounting for 10.7%, 4.9%, 1.9% and 0.9% of isolates, respectively. In conclusion, this work reports epidemiological data on adenoviral conjunctivitis from a region where such information is very scarce and contributes to a better knowledge of the worldwide distribution of causative genotypes. It also presents an approach for the identification of circulating HAdV in the country and demonstrates the importance of molecular tools for both detection and identification of genotypes, which allow rapid virological investigation, especially during epidemics. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  18. Adenoviral short hairpin RNA therapy targeting phosphodiesterase 5a relieves cardiac remodeling and dysfunction following myocardial infarction.

    PubMed

    Li, Longhu; Haider, Husnain Kh; Wang, Linlin; Lu, Gang; Ashraf, Muhammad

    2012-05-15

    We previously showed that treatment with tadalafil, a long-acting phosphodiesterase-5a (PDE5a) inhibitor, effectively prevented adverse left ventricular (LV) remodeling of the infarcted heart. We hypothesized that short-hairpin RNA (shRNA) therapy targeting PDE5a would simulate the effects of pharmacological intervention for treatment of postinfarction LV remodeling and dysfunction. Experimental model of myocardial infarction was developed in female mice by permanent ligation of left coronary artery. Immediately after that, an adenoviral vector encoding for shRNA sequence targeting PDE5a (Ad-shPDE5a) was injected intramyocardially, which specifically inhibited PDE5a in the heart. Four weeks later, Ad-shPDE5a treated mice showed significant mitigation of the left ventricle (LV) dilatation and dysfunction as indicated by smaller LV cavity and more preserved ejection fraction and fractional shortening. Infarction size and fibrosis were significantly reduced in Ad-shPDE5a-treated mice. Additionally, more salvaged cardiomyocytes, significantly reduced collagen contents, and higher blood vessel density were observed in Ad-shPDE5a-treated mice. The cytoprotective effects of Ad-shPDE5a were demonstrated in vitro in Ad-shPDE5a transfected cardiomyocytes cultured under oxygen glucose deprivation. Among downstream mediators of PDE5a signaling, cyclic GMP (cGMP) and cGMP-dependent protein kinase G (PKG) were activated with concomitant reduction in caspase-3 activity. However, no significant change in PKA and cAMP activities were observed in Ad-shPDE5a-treated hearts. Inhibition with shRNA improved cardiac remodeling and dysfunction by reducing infarction size and cardiac fibrosis and increased cGMP and PKG activity. These findings suggest that PDE5 inhibition with Ad-shPDE5a is a novel approach for treatment of myocardial infarction.

  19. Adenoviral short hairpin RNA therapy targeting phosphodiesterase 5a relieves cardiac remodeling and dysfunction following myocardial infarction

    PubMed Central

    Li, Longhu; Haider, Husnain Kh.; Wang, Linlin; Lu, Gang

    2012-01-01

    We previously showed that treatment with tadalafil, a long-acting phosphodiesterase-5a (PDE5a) inhibitor, effectively prevented adverse left ventricular (LV) remodeling of the infarcted heart. We hypothesized that short-hairpin RNA (shRNA) therapy targeting PDE5a would simulate the effects of pharmacological intervention for treatment of postinfarction LV remodeling and dysfunction. Experimental model of myocardial infarction was developed in female mice by permanent ligation of left coronary artery. Immediately after that, an adenoviral vector encoding for shRNA sequence targeting PDE5a (Ad-shPDE5a) was injected intramyocardially, which specifically inhibited PDE5a in the heart. Four weeks later, Ad-shPDE5a treated mice showed significant mitigation of the left ventricle (LV) dilatation and dysfunction as indicated by smaller LV cavity and more preserved ejection fraction and fractional shortening. Infarction size and fibrosis were significantly reduced in Ad-shPDE5a-treated mice. Additionally, more salvaged cardiomyocytes, significantly reduced collagen contents, and higher blood vessel density were observed in Ad-shPDE5a-treated mice. The cytoprotective effects of Ad-shPDE5a were demonstrated in vitro in Ad-shPDE5a transfected cardiomyocytes cultured under oxygen glucose deprivation. Among downstream mediators of PDE5a signaling, cyclic GMP (cGMP) and cGMP-dependent protein kinase G (PKG) were activated with concomitant reduction in caspase-3 activity. However, no significant change in PKA and cAMP activities were observed in Ad-shPDE5a-treated hearts. Inhibition with shRNA improved cardiac remodeling and dysfunction by reducing infarction size and cardiac fibrosis and increased cGMP and PKG activity. These findings suggest that PDE5 inhibition with Ad-shPDE5a is a novel approach for treatment of myocardial infarction. PMID:22447941

  20. Phase polymorphism of novel [Ru(NH{sub 3}){sub 6}](ClO{sub 4}){sub 3}—Comparison with [Ru(NH{sub 3}){sub 6}](BF{sub 4}){sub 3}. Part II

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

    Dołęga, Diana, E-mail: dolega@chemia.uj.edu.pl; Mikuli, Edward, E-mail: mikuli@chemia.uj.edu.pl; Górska, Natalia, E-mail: natalia.gorska@uj.edu.pl

    2013-08-15

    [Ru(NH{sub 3}){sub 6}](ClO{sub 4}){sub 3} undergoes two phase transitions at: T{sub C1}=290.3 K and T{sub C2}=74.8 K , thus exhibits three crystalline phases in the temperature range of 5–310 K. For the detected phase transitions, thermal effects were determined. Fourier transform far- and middle-infrared spectra (FT-FIR and FT-MIR), recorded at 8–350 K, suggest that reorientational motions of the NH{sub 3} ligands are very fast (τ{sub R}≈10{sup −12} s above T{sub C1}) and are significantly slowed down below T{sub C2}. X-ray single crystal diffraction (XRSCD) measurements revealed that in the high temperature phase (above T{sub C1}) the compound belongs to themore » cubic Fm3{sup ¯}m (No. 225) space group, whereas in the intermediate phase the unit cell parameter doubles and the space group is Ia3{sup ¯}(No. 206). {sup 1}H NMR studies revealed that the following reorientational motions are liberated during heating: three-fold reorientation of NH{sub 3} ligands, three-fold reorientation of the entire [Ru(NH{sub 3}){sub 6}]{sup 3+} cation, and isotropic reorientation of this cation. In the high temperature phase I the cations perform isotropic reorientations with the estimated activation energy equal to ca. 30.1 kJ mol{sup −1}. Comparison with adequate results obtained earlier for [Ru(NH{sub 3}){sub 6}](BF{sub 4}){sub 3} and for other similar compounds was made and general regularities were drawn. - Graphical abstract: Molar heat capacities obtained for [Ru(NH{sub 3}){sub 6}](ClO{sub 4}){sub 3} at temperatures between 5 and 310 K. Molecular structure of [Ru(NH{sub 3}){sub 6}]{sup 3+} and two types of ClO{sub 4}{sup -} varying in dynamic disorder at 293 K. Highlights: • Two solid phase transitions have been found in [Ru(NH{sub 3}){sub 6}](ClO{sub 4}){sub 3}. • The transitions are triggered by the hydrogen bond interactions between NH{sub 3} ligands and ClO{sub 4}{sup −} anions. • The complex is a highly dynamically disordered crystal across a

  1. Stimulation of a Ca sup 2+ -dependent protein kinase by G sub M1 ganglioside in nerve growth factor-treated PC12 cells

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

    Hilbush, B.S.; Levine, J.M.

    1991-07-01

    The authors have investigated the ability of exogenous gangliosides to modulate nerve growth factor (NGF) signal transduction in PC12 cells. The effects of exogenous ganglioside G{sub M1} on multiple protein kinase activities were assayed by analyzing site-specific serine phosphorylation of tyrosine hydroxylase (TyrOHase) by two-dimensional phosphopeptide mapping. In the presence of NGF, exogenous G{sub M1} increased {sup 32}P incorporation into TyrOHase phosphopeptide T2, a Ca{sup 2+}/calmodulin-dependent protein kinase substrate whose phosphorylation is not normally affected by NGF treatment. In the absence of NGF, G{sub M1} treatment had no significant effects on TyrOHase phosphorylation. The removal of extracellular Ca{sup 2+} ormore » blockade of dihydropyridine-sensitive Ca{sup 2+} channels prevented the G{sub M1}-induced increases in {sup 32}P incorporation into phosphopeptide T2. Exogenous G{sub M1} also potentiated K{sup +} depolarization-induced increases in the phosphorylation of TyrOHase. These results suggest that the stimulatory effects of exogenous G{sub M1} ganglioside on NGF actions may be due to its ability to potentiate a Ca{sup 2+}-dependent signaling pathway.« less

  2. Optimization and physicochemical characterization of a cationic lipid-phosphatidylcholine mixed emulsion formulated as a highly efficient vehicle that facilitates adenoviral gene transfer.

    PubMed

    Kim, Soo-Yeon; Lee, Sang-Jin; Kim, Jin-Ki; Choi, Han-Gon; Lim, Soo-Jeong

    2017-01-01

    Cationic lipid-based nanoparticles enhance viral gene transfer by forming electrostatic complexes with adenoviral vectors. We recently demonstrated the superior complexation capabilities of 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) emulsion compared with a liposomal counterpart but the cytotoxicity of DOTAP emulsions remained a challenge. The present study is aimed at formulating an emulsion capable of acting as a highly effective viral gene transfer vehicle with reduced cytotoxicity and to physicochemically characterize the structures of virus-emulsion complexes in comparison with virus-liposome complexes when the only difference between emulsions and liposomes was the presence or absence of inner oil core. The emulsion formulation was performed by 1) reducing the content of DOTAP while increasing the content of zwitterionic lipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and 2) optimizing the oil content. The complexation capability of formulated DOTAP:DMPC mixed emulsions was similar to those of emulsions containing DOTAP alone while displaying significantly lower cytotoxicity. The complexation capabilities of the DOTAP:DMPC mixed emulsion were serum-compatible and were monitored in a variety of cell types, whereas its liposomal counterpart was totally ineffective. Characterization by scanning electron microscopy, transmission electron microscopy, atomic force microscopy, and dynamic light scattering studies indicated that the optimized emulsions spontaneously surrounded the virus particles to generate emulsions that encapsulated the viral particles, whereas viral particles merely attached to the surfaces of the counterpart liposomes to form multiviral aggregates. Overall, these studies demonstrated that optimized DOTAP:DMPC mixed emulsions are potentially useful for adenoviral gene delivery due to less cytotoxicity and the unique ability to encapsulate the viral particle, highlighting the importance of nanoparticle formulation.

  3. Optimization and physicochemical characterization of a cationic lipid-phosphatidylcholine mixed emulsion formulated as a highly efficient vehicle that facilitates adenoviral gene transfer

    PubMed Central

    Kim, Soo-Yeon; Lee, Sang-Jin; Kim, Jin-Ki; Choi, Han-Gon; Lim, Soo-Jeong

    2017-01-01

    Cationic lipid-based nanoparticles enhance viral gene transfer by forming electrostatic complexes with adenoviral vectors. We recently demonstrated the superior complexation capabilities of 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) emulsion compared with a liposomal counterpart but the cytotoxicity of DOTAP emulsions remained a challenge. The present study is aimed at formulating an emulsion capable of acting as a highly effective viral gene transfer vehicle with reduced cytotoxicity and to physicochemically characterize the structures of virus-emulsion complexes in comparison with virus–liposome complexes when the only difference between emulsions and liposomes was the presence or absence of inner oil core. The emulsion formulation was performed by 1) reducing the content of DOTAP while increasing the content of zwitterionic lipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and 2) optimizing the oil content. The complexation capability of formulated DOTAP:DMPC mixed emulsions was similar to those of emulsions containing DOTAP alone while displaying significantly lower cytotoxicity. The complexation capabilities of the DOTAP:DMPC mixed emulsion were serum-compatible and were monitored in a variety of cell types, whereas its liposomal counterpart was totally ineffective. Characterization by scanning electron microscopy, transmission electron microscopy, atomic force microscopy, and dynamic light scattering studies indicated that the optimized emulsions spontaneously surrounded the virus particles to generate emulsions that encapsulated the viral particles, whereas viral particles merely attached to the surfaces of the counterpart liposomes to form multiviral aggregates. Overall, these studies demonstrated that optimized DOTAP:DMPC mixed emulsions are potentially useful for adenoviral gene delivery due to less cytotoxicity and the unique ability to encapsulate the viral particle, highlighting the importance of nanoparticle formulation. PMID:29070949

  4. Structure and superconductivity in (Bi{sub 0.35}Cu{sub 0.65})Sr{sub 2}YCu{sub 2}O{sub 7} and related materials

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

    Jennings, R.A.; Williams, S.P.; Greaves, C.

    1994-12-31

    The recently reported (Bi/Cu)Sr{sub 2}YCu{sub 2}O{sub 7} phase has been studied by time of flight powder neutron diffraction. The proposed 1212 structure has been confirmed and refinements have shown the oxygen in the (Bi/Cu)O layer is displaced by 0.78{angstrom} from the ideal (1/2,1/2,0) site (P4/mmm space group) along (100). Bond Valence Sum calculations have suggested oxidation states of Bi{sup 5+} and Cu{sup 2+} for the cations in the (Bi/Cu)O layers. The material is non-superconducting and all attempts to induce superconductivity have been unsuccessful. Work on the related material (Ce/Cu)Sr{sub 2}YCu{sub 2}O{sub y} has shown the ideal Ce content to bemore » 0.5 Ce per formula unit. The introduction of Ba (10%) onto the Sr site dramatically increases phase stability and also induces superconductivity (62K).« less

  5. Rietveld refinement and ionic conductivity of Ca{sub 8.4}Bi{sub 1.6}(PO{sub 4}){sub 6}O{sub 1.8}

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

    Tmar Trabelsi, I., E-mail: ilhem_tmar@yahoo.fr; Madani, A.; Mercier, A.M.

    2013-01-15

    The structure of Ca{sub 8.4}Bi{sub 1.6}(PO{sub 4}){sub 6}O{sub 1.8}, isostructural with Fluoroapatite, was determined by X-ray powder diffraction methods. The results of Rietveld refinement revealed that the formula of this compound is [Ca{sub 4}]{sup 4f}[Ca{sub 4.4}Bi{sub 1.6}]{sup 6h}(PO{sub 4}){sub 6}[O{sub 1.8}]{sup 2a}, space group P63/m (a=9.468 (3) A, c=6.957 (3) A). A total substitution of Bi{sup 3+} ions in the (6h) sites was related particularly to the high polarizability of the Bi{sup 3+} ion compared to Ca{sup 2+}. The observed frequencies in the Raman and infrared spectra were explained and discussed on the basis of unit-cell group analyses and inmore » comparison with Fluoroapatite and other oxyapatites. The ionic conductivity over a wide range of temperature was investigated according to the complex impedance method. The highest overall conductivity values were found at {sigma}{sub 700 Degree-Sign C} =5.03 Multiplication-Sign 10{sup -7} S cm{sup -1} and E{sub a}=0.50 eV. - Graphical abstract: The final Rietveld refinement plot of the Ca{sub 8.4}Bi{sub 1.6} (PO{sub 4}){sub 6}O{sub 1.8}. Highlights: Black-Right-Pointing-Pointer The Rietveld refinement revealed that the formula of this compound is Ca{sub 8.4}Bi{sub 1.6}(PO{sub 4}){sub 6}O{sub 1.8}. Black-Right-Pointing-Pointer Vibrational spectroscopy supports the high symmetry P63/m space group for this apatite. Black-Right-Pointing-Pointer This apatite contained channels where oxygen ions were located in 2a sites. Black-Right-Pointing-Pointer The possibility of anionic conduction along these channels was considered.« less

  6. Solvothermal syntheses, crystal structures, and properties of lanthanide(III) thioarsenates [Ln(dien){sub 2}(μ-1κ,2κ{sup 2}-AsS{sub 4})]{sub n} (Ln==Sm, Eu, Gd) and [Ln(dien){sub 2}(1κ{sup 2}-AsS{sub 4})] (Ln==Tb, Dy, Ho)

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

    Wang, Fang; Tang, Chunying; Chen, Ruihong

    2013-10-15

    Solvothermal reactions of Ln{sub 2}O{sub 3}, As and S in diethylenetriamine (dien) at 170 °C for 6 days afforded two structural types of lanthanide thioarsenates with the general formulae [Ln(dien){sub 2}(μ-1κ,2κ{sup 2}-AsS{sub 4})]{sub n} [Ln=Sm(1), Eu(2), Gd(3)] and [Ln(dien){sub 2}(1κ{sup 2}-AsS{sub 4})] [Ln=Tb(4), Dy(5), Ho(6)]. The Ln{sub 2}O{sub 3} oxides were converted to [Ln(dien){sub 2}]{sup 3+} complex units in the solvothermal reactions. The As atom binds four S atoms, forming a tetrahedral AsS{sub 4} unit. In 1−3, the AsS{sub 4} units interconnect the [Ln(dien){sub 2}]{sup 3+} cations via Ln−S bonds as tridentate μ-1κ,2κ{sup 2}-AsS{sub 4} bridging ligands, resulting in themore » neutral coordination polymers [Ln(dien){sub 2}(μ-1κ,2κ{sup 2}-AsS{sub 4})]{sub n} (Ln1). In 4−6, the AsS{sub 4} units coordinate with the Ln{sup 3+} ion of [Ln(dien){sub 2}]{sup 3+} as 1κ{sup 2}-AsS{sub 4} chelating ligands to form neutral coordination compounds [Ln(dien){sub 2}(1κ{sup 2}-AsS{sub 4})] (Ln2). The Ln{sup 3+} ions are in nine- and eight-coordinated environments in Ln1 and Ln2, respectively. The formation of Ln1 and Ln2 is related with ionic size of the Ln{sup 3+} ions. Optical absorption spectra showed that 1−6 have potential use as semiconductors with the band gaps in the range 2.18−3.21 eV. - Graphical abstract: Two types of Ln-thioarsenates [Ln(dien){sub 2}(μ-1κ,2κ{sup 2}-AsS{sub 4})]{sub n} and [Ln(dien){sub 2}(1κ{sup 2}-AsS{sub 4})] were prepared by solvothermal methods and the soft Lewis basic AsS{sub 4}{sup 3–} ligand to Ln(III) centers with polyamine co-ligand was obtained. Display Omitted - Highlights: • Lanthanide thioarsenates were prepared by solvothermal methods. • The soft Lewis basic AsS{sub 4} ligand coordinate Ln{sup 3+} ions with coexistence polyamine ligands. • Two structural types of Ln-thioarsenates with structural turnover at Tb were obtained along Ln series. • The Ln-thioarsenates are potential

  7. From Ba{sub 3}Ta{sub 5}O{sub 14}N to LaBa{sub 2}Ta{sub 5}O{sub 13}N{sub 2}: Decreasing the optical band gap of a photocatalyst

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

    Anke, B.; Bredow, T.; Pilarski, M.

    Yellow LaBa{sub 2}Ta{sub 5}O{sub 13}N{sub 2} was successfully synthesized as phase-pure material crystallizing isostructurally to previously reported Ba{sub 3}Ta{sub 5}O{sub 14}N and mixed-valence Ba{sub 3}Ta{sup V}{sub 4}Ta{sup IV}O{sub 15}. The electronic structure of LaBa{sub 2}Ta{sub 5}O{sub 13}N{sub 2} was studied theoretically with the range-separated hybrid method HSE06. The most stable structure was obtained when lanthanum was placed on 2a and nitrogen on 4h sites confirming Pauling's second rule. By incorporating nitrogen, the measured band gap decreases from ∼3.8 eV for the oxide via 2.74 eV for Ba{sub 3}Ta{sub 5}O{sub 14}N to 2.63 eV for the new oxide nitride, giving risemore » to an absorption band well in the visible-light region. Calculated fundamental band gaps confirm the experimental trend. The atom-projected density of states has large contributions from N2p orbitals close to the valence band edge. These are responsible for the observed band gap reduction. Photocatalytic hydrogen formation was investigated and compared with that of Ba{sub 3}Ta{sub 5}O{sub 14}N revealing significantly higher activity for LaBa{sub 2}Ta{sub 5}O{sub 13}N{sub 2} under UV-light. - Graphical abstract: X-ray powder diffraction pattern of LaBa{sub 2}Ta{sub 5}O{sub 13}N{sub 2} with the results of the Rietveld refinements. Inset: Unit cell of LaBa{sub 2}Ta{sub 5}O{sub 13}N{sub 2} and polyhedral representation of the crystal structure. - Highlights: • Synthesis of a new oxide nitride LaBa{sub 2}Ta{sub 5}O{sub 13}N{sub 2}. • Refinement of the crystal structure. • Quantum chemical calculations provided band gap close to the measured value. • New phase shows a higher photocatalytic H{sub 2} evolution rate compared to prior tested Ba{sub 3}Ta{sub 5}O{sub 14}N.« less

  8. Experimental and ab initio studies on sub-lattice ordering and magnetism in Co{sub 2}Fe(Ge{sub 1−x}Si{sub x}) alloys

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

    Deka, Bhargab; Kundu, Ashis; Ghosh, Subhradip

    2015-10-07

    Crystallographic and magnetic properties of bulk Co{sub 2}Fe(Ge{sub 1−x}Si{sub x}) alloys with 0 ≤ x ≤ 1, synthesized by arc melting method, have been studied. Co{sub 2}FeSi alloy has been found to crystallize with L2{sub 1} structure, but the super-lattice peaks are absent in the X-ray diffraction patterns of alloys containing high Ge concentration. Unit cell volume of this series of alloys decreased from 185.2 to 178.5 Å{sup 3} as Si content was increased from 0 to 1.00. All alloy compositions exhibit ferromagnetic behavior with a high Curie temperature (T{sub C}). T{sub C} showed a systematic variation with x. A comparison between the valuesmore » of saturation magnetization (M{sub s}) and effective moment per magnetic atom p{sub c} estimated from the temperature dependent susceptibility data above T{sub C}, shows that the alloys have half-metallic character. The alloy with x = 0 follows Slater-Pauling (S-P) rule with M{sub s} of 5.99μ{sub B}. However, M{sub s} for the alloy with x = 1.00 was found to be 5.42μ{sub B}, which is lower than the value of 6.0μ{sub B} predicted by S-P rule. Since atomic disorder is known to affect the M{sub s} and electronic structure of these alloys, ab initio calculations were carried out to explain the deviation in observed M{sub s} from S-P rule prediction and the half-metallic character of the alloys. Ab initio calculations reveal that alloys with L2{sub 1} structure have M{sub s} value as predicted by S-P rule. However, introduction of 12.5% DO{sub 3} disorder, which occurs due to swapping of Co and Fe atoms in the unit cell, decreases M{sub s} of alloys with x > 0 from the S-P prediction to values obtained experimentally. The results analyzed from the view point of electronic structure of the alloys in different ordered states bring out the influence of disorder on the observed magnetic properties of these technologically important alloys.« less

  9. Detection and phylogenetic analysis of a new adenoviral polymerase gene in reptiles in Korea.

    PubMed

    Bak, Eun-Jung; Jho, Yeonsook; Woo, Gye-Hyeong

    2018-06-01

    Over a period of 7 years (2004-2011), samples from 34 diseased reptiles provided by local governments, zoos, and pet shops were tested for viral infection. Animals were diagnosed based on clinical signs, including loss of appetite, diarrhea, rhinorrhea, and unexpected sudden death. Most of the exotic animals had gastrointestinal problems, such as mucosal redness and ulcers, while the native animals had no clinical symptoms. Viral sequences were found in seven animals. Retroviral genes were amplified from samples from five Burmese pythons (Python molurus bivittatus), an adenovirus was detected in a panther chameleon (Furcifer pardalis), and an adenovirus and a paramyxovirus were detected in a tropical girdled lizard (Cordylus tropidosternum). Phylogenetic analysis of retroviruses and paramyxoviruses showed the highest sequence identity to both a Python molurus endogenous retrovirus and a Python curtus endogenous retrovirus and to a lizard isolate, respectively. Partial sequencing of an adenoviral DNA polymerase gene from the lizard isolate suggested that the corresponding virus was a novel isolate different from the reference strain (accession no. AY576677.1). The virus was not isolated but was detected, using molecular genetic techniques, in a lizard raised in a pet shop. This animal was also coinfected with a paramyxovirus.

  10. Glasses of the As/sub 2/S/sub 3/-T1/sub 2/S system

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

    Gutenev, M.S.

    1986-08-01

    A dielcometric study of (AsS /SUB 1.5/ ) /SUB 1-x/ (TiS /SUB 0.5/ ) /SUB x/ (0 is less than or equal to x is less than or equal to 0.61) glasses was carried out. Glassforming alloys were prepared in thin-walled quartz ampules by rapid cooling from 700 C in air. The methods of determination of permittivity, refractive index, and density, the values of which are shown here, have been previously discussed. The molar infrared polarizability is calculated from the experimental data previously gathered, and the concentration dependence is shown. In this paper, the presence of chemical atomic order inmore » T1AsS/sub 2/ glass described by TISAsS /SUB 2/2/ structural units was experimentally proved. An assumption was made of strong mutual influence of T1AsS/sub 2/ and AsS /SUB 1.5/ complexes caused by coordination of thallium with bridging sulfur atoms.« less

  11. [Role of G-protein alpha sub-units in the morphogenic processes of filamentous Ascomycota fungi].

    PubMed

    García-Rico, Ramón O; Fierro, Francisco

    The phylum Ascomycota comprises about 75% of all the fungal species described, and includes species of medical, phytosanitary, agricultural, and biotechnological importance. The ability to spread, explore, and colonise new substrates is a feature of critical importance for this group of organisms. In this regard, basic processes such as conidial germination, the extension of hyphae and sporulation, make up the backbone of development in most filamentous fungi. These processes require specialised morphogenic machinery, coordinated and regulated by mechanisms that are still being elucidated. In recent years, substantial progress has been made in understanding the role of the signalling pathway mediated by heterotrimericG proteins in basic biological processes of many filamentous fungi. This review focuses on the role of the alpha subunits of heterotrimericG proteins in the morphogenic processes of filamentous Ascomycota. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  12. Ex vivo adenoviral gene transfer of constitutively activated STAT3 reduces post-transplant liver injury and promotes regeneration in a 20% rat partial liver transplant model.

    PubMed

    Huda, Kamrul A S M; Guo, Lei; Haga, Sanae; Murata, Hiroshi; Ogino, Tetsuya; Fukai, Moto; Yagi, Takahito; Iwagaki, Hiromi; Tanaka, Noriaki; Ozaki, Michitaka

    2006-05-01

    Signal transducer and activator of transcription-3 (STAT3) is one of the most important transcription factors for liver regeneration. This study was designed to examine the effects of constitutively activated STAT3 (STAT3-C) on post-transplant liver injury and regeneration in a rat 20% partial liver transplant (PLTx) model by ex vivo adenoviral gene transfer. Adenovirus encoding the STAT3-C gene was introduced intraportally into liver grafts and clamped for 30 min during cold preservation. After orthotopic PLTx, liver graft/body weights and serum biochemistry were monitored, and both a histological study and DNA binding assay were performed. STAT3-C protein expression and its binding to DNA in the liver graft were confirmed by Western blotting and electrophoretic mobility shift assay (EMSA), respectively. This treatment modality promoted post-Tx liver regeneration effectively and rapidly. The serum levels of alanine aminotransferase/aspartate aminotransferase (AST/ALT) and bilirubin decreased in rats with STAT3-C. However, albumin (a marker of liver function) did not. Ex vivo gene transfer of STAT3-C to liver grafts reduced post-Tx injury and promoted liver regeneration. Thus, the activation of STAT3 in the liver graft may be a potentially effective clinical strategy for improving the outcome of small-for-size liver transplantation.

  13. Lattice thermal expansion of the solid solutions (La{sub 1−x}Sm{sub x}){sub 2}Ce{sub 2}O{sub 7}

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

    Wu, Hongdan; Lei, Xinrong; Zhang, Jinhua, E-mail: jhzhang1212@126.com

    2014-09-15

    Highlights: • Sm-doped La{sub 2}Ce{sub 2}O{sub 7} was prepared by the coprecipitation–calcination method. • In situ HT-XRD measurements revealed that is much stable than 8YSZ. • Its thermal expansion is better than 8YSZ. - Abstract: A series of solid solutions with the general formula (La{sub 1−x}Sm{sub x}){sub 2}Ce{sub 2}O{sub 7} (0.0 ≤ x ≤ 1.0) were prepared by the coprecipitation–calcination method. The products obtained were characterized by powder X-ray diffraction for phase purity. It was observed that La{sup 3+} and Sm{sup 3+} can form complete solid solution in (La,Sm){sub 2}Ce{sub 2}O{sub 7} with defect-fluorite-type phase. The unit cell parameters ofmore » these solutions were calculated by a least squares method and the lattice parameters decreased linearly as x increased. The lattice thermal expansion behavior of (La{sub 1−x}Sm{sub x}){sub 2}Ce{sub 2}O{sub 7} (0.0 ≤ x ≤ 1.0) was investigated by high-temperature X-ray diffraction in the temperature range 298–1623 K. The lattice parameters a{sub T} of all the solutions at different temperature can be expressed as a{sub T} = a + bT + cT{sup 2}. As x < 1, the thermal expansion has a sudden decrease at ca. 473 K. The coefficients of lattice thermal expansion of Sm{sub 2}Ce{sub 2}O{sub 7} were 10.2–13.6 × 10{sup −6} K{sup −1} from 298 to 1623 K, and without the thermal contraction at low temperature. The materials show positive or negative thermal expansion due to the asymmetric anharmonic vibration.« less

  14. Planar dicyclic B{sub 6}S{sub 6}, B{sub 6}S{sub 6}{sup −}, and B{sub 6}S{sub 6}{sup 2−} clusters: Boron sulfide analogues of naphthalene

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

    Li, Da-Zhi; Bai, Hui; Ou, Ting

    2015-01-07

    Inorganic analogues of hydrocarbons or polycyclic aromatic hydrocarbons (PAHs) are of current interest in chemistry. Based upon global structural searches and B3LYP and CCSD(T) calculations, we present herein the perfectly planar dicyclic boron sulfide clusters: D{sub 2h} B{sub 6}S{sub 6} (1, {sup 1}A{sub g}), D{sub 2h} B{sub 6}S{sub 6}{sup −} (2, {sup 2}B{sub 3u}), and D{sub 2h} B{sub 6}S{sub 6}{sup 2−} (3, {sup 1}A{sub g}). These are the global minima of the systems, being at least 0.73, 0.81, and 0.53 eV lower in energy, respectively, than their alternative isomers at the CCSD(T) level. The D{sub 2h} structures feature twin B{submore » 3}S{sub 2} five-membered rings, which are fused together via a B{sub 2} unit and terminated by two BS groups. Bonding analyses show that the closed-shell B{sub 6}S{sub 6}{sup 2−} (3) cluster possesses 10 delocalized π electrons, closely analogous to the bonding pattern of the aromatic naphthalene C{sub 10}H{sub 8}. The B{sub 6}S{sub 6}{sup −} (2) and B{sub 6}S{sub 6} (1) species are readily obtained upon removal of one or two π electrons from B{sub 6}S{sub 6}{sup 2−} (3). The results build a new analogous relationship between boron sulfide clusters and their PAH counterparts. The B{sub 6}S{sub 6}{sup −} (2) monoanion and B{sub 6}S{sub 6}{sup 2−} (3) dianion can be effectively stabilized in neutral LiB{sub 6}S{sub 6} and Li{sub 2}B{sub 6}S{sub 6} salts, respectively.« less

  15. Local structure of Rb{sub 2}Li{sub 4}(SeO{sub 4}){sub 3}{center_dot}2H{sub 2}O by the modeling of X-ray diffuse scattering - from average-structure to microdomain model

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

    Komornicka, Dorota; Wolcyrz, Marek, E-mail: m.wolcyrz@int.pan.wroc.pl; Pietraszko, Adam

    2012-08-15

    Local structure of dirubidium tetralithium tris(selenate(VI)) dihydrate - Rb{sub 2}Li{sub 4}(SeO{sub 4}){sub 3}{center_dot} 2H{sub 2}O has been determined basing on the modeling of X-ray diffuse scattering. The origin of observed structured diffuse streaks is SeO{sub 4} tetrahedra switching between two alternative positions in two quasi-planar layers existing in each unit cell and formation of domains with specific SeO{sub 4} tetrahedra configuration locally fulfilling condition for C-centering in the 2a Multiplication-Sign 2b Multiplication-Sign c superstructure cell. The local structure solution is characterized by a uniform distribution of rather large domains (ca. thousand of unit cells) in two layers, but also monodomainsmore » can be taken into account. Inside a single domain SeO{sub 4} tetrahedra are ordered along ab-diagonal forming two-string ribbons. Inside the ribbons SeO{sub 4} and LiO{sub 4} tetrahedra share the oxygen corners, whereas ribbons are bound to each other by a net of hydrogen bonds and fastened by corner sharing SeO{sub 4} tetrahedra of the neighboring layers. - Graphical abstract: Experimental sections of the reciprocal space showing diffraction effects observed for RLSO. Bragg spots are visible on sections with integer indices (1 kl section - on the left), streaks - on sections with fractional ones (1.5 kl section - on the right). At the center: resulting local structure of the A package modeled as a microdomain: two-string ribbons of ordered oxygen-corners-sharing SeO{sub 4} and LiO{sub 4} terahedra extended along ab-diagonal are seen; ribbons are bound by hydrogen bonds (shown in pink); the multiplied 2a Multiplication-Sign 2b unit cell is shown. Highlights: Black-Right-Pointing-Pointer X-ray diffuse scattering in RLSO was registered and modeled. Black-Right-Pointing-Pointer The origin of diffuse streaks is SeO{sub 4} tetrahedra switching in two structure layers. Black-Right-Pointing-Pointer The local structure is characterized by a

  16. Negative thermal expansion correlated with polyhedral movements and distortions in orthorhombic Y{sub 2}Mo{sub 3}O{sub 12}

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

    Wang, Lei; Wang, Fei; Yuan, Peng-Fei

    Graphical abstract: Our work confirms the negative thermal expansion (NTE) behavior of the orthorhombic Y{sub 2}Mo{sub 3}O{sub 12} in this range 0–1000 K. The orthorhombic Y{sub 2}Mo{sub 3}O{sub 12} has an open framework structure where MoO{sub 4} tetrahedra and YO{sub 6} octahedra are connected by oxygen atoms. The previous mechanisms for the NTE behavior of orthorhombic Y{sub 2}Mo{sub 3}O{sub 12} are that the translational mode (see (b)) of the O bridge atoms in Y-O-Mo linkages will cause the linkages to be bent, reducing the space between polyhedra and making the volumetric shrinkage. Furthermore, the internal polyhedral distortions have been reportedmore » experimentally. It is necessary to reveal the relationship between NTE and polyhedral movements, distortions. From the vibrational properties, we get that the different vibrational eigenvectors of oxygen atoms relative to Y or Mo atoms can lead internal polyhedra to distort unevenly (see (c)). Herein, an extended 3D model of the connected unit YO{sub 6}-MoO{sub 4} based on the Y-O-Mo linkage is proposed (see (a)). It presents a simultaneous dynamic process, i.e. the YO{sub 6} octahedra and MoO{sub 4} tetrahedra distort unevenly, along with both polyhedra being closer which makes the volumetric contraction. This model is helpful to improve the mechanisms of NTE and may be applied in the whole A{sub 2}M{sub 3}O{sub 12} family. - Highlights: • The NTE properties of Y{sub 2}Mo{sub 3}O{sub 12} are confirmed using a first-principles calculation. • The optical branch with the lowest frequency is most responsible for the NTE. • The relationship between NTE and polyhedral movements, distortions is elucidated. • An extended 3D model of the connected unit YO{sub 6}-MoO{sub 4} is proposed. - Abstract: The internal polyhedral distortions have been reported experimentally in orthorhombic Y{sub 2}Mo{sub 3}O{sub 12} as a negative thermal expansion (NTE) material. To reveal the relationship between NTE and

  17. Fall risk as a function of time after admission to sub-acute geriatric hospital units.

    PubMed

    Rapp, Kilian; Ravindren, Johannes; Becker, Clemens; Lindemann, Ulrich; Jaensch, Andrea; Klenk, Jochen

    2016-10-07

    There is evidence about time-dependent fracture rates in different settings and situations. Lacking are data about underlying time-dependent fall risk patterns. The objective of the study was to analyse fall rates as a function of time after admission to sub-acute hospital units and to evaluate the time-dependent impact of clinical factors at baseline on fall risk. This retrospective cohort study used data of 5,255 patients admitted to sub-acute units in a geriatric rehabilitation clinic in Germany between 2010 and 2014. Falls, personal characteristics and functional status at admission were extracted from the hospital information system. The rehabilitation stay was divided in 3-day time-intervals. The fall rate was calculated for each time-interval in all patients combined and in subgroups of patients. To analyse the influence of covariates on fall risk over time multivariate negative binomial regression models were applied for each of 5 time-intervals. The overall fall rate was 10.2 falls/1,000 person-days with highest fall risks during the first week and decreasing risks within the following weeks. A particularly pronounced risk pattern with high fall risks during the first days and decreasing risks thereafter was observed in men, disoriented people, and people with a low functional status or impaired cognition. In disoriented patients, for example, the fall rate decreased from 24.6 falls/1,000 person-days in day 2-4 to about 13 falls/1,000 person-days 2 weeks later. The incidence rate ratio of baseline characteristics changed also over time. Fall risk differs considerably over time during sub-acute hospitalisation. The strongest association between time and fall risk was observed in functionally limited patients with high risks during the first days after admission and declining risks thereafter. This should be considered in the planning and application of fall prevention measures.

  18. The Bi{sub 2}O{sub 3}–Fe{sub 2}O{sub 3}–Sb{sub 2}O{sub 5} system phase diagram refinement, Bi{sub 3}FeSb{sub 2}O{sub 11} structure peculiarities and magnetic properties

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

    Egorysheva, A.V., E-mail: anna_egorysheva@rambler.ru; Ellert, O.G.; Gajtko, O.M.

    2015-05-15

    The refinement of the Bi{sub 2}O{sub 3}–Fe{sub 2}O{sub 3}–Sb{sub 2}O{sub 5} system phase diagram has been performed and the existence of the two ternary compounds has been confirmed. The first one with a pyrochlore-type structure (sp. gr. Fd 3-barm) exists in the wide solid solution region, (Bi{sub 2−x}Fe{sub x})Fe{sub 1+y}Sb{sub 1−y}O{sub 7±δ}, where x=0.1–0.4 and y=−0.13–0.11. The second one, Bi{sub 3}FeSb{sub 2}O{sub 11}, corresponds to the cubic KSbO{sub 3}-type structure (sp. gr. Pn 3-bar) with unit cell parameter a=9.51521(2) Å. The Rietveld structure refinement showed that this compound is characterized by disordered structure. The Bi{sub 3}FeSb{sub 2}O{sub 11} factor groupmore » analysis has been carried out and a Raman spectrum has been investigated. According to magnetization measurements performed at the temperature range 2–300 K it may be concluded that the Bi{sub 3}FeSb{sub 2}O{sub 11} magnetic properties can be substantially described as a superposition of strong short-range antiferromagnetic exchange interactions realizing inside the [(FeSb{sub 2})O{sub 9}] 3D-framework via different pathways. - Graphical abstract: The refinement of the Bi{sub 2}O{sub 3}–Fe{sub 2}O{sub 3}–Sb{sub 2}O{sub 5} system phase diagram has been performed and the existence of the solid solution with a pyrochlore-type structure (sp. gr. Fd 3-barm) and Bi{sub 3}FeSb{sub 2}O{sub 11}, correspond of the cubic KSbO{sub 3}-type structure (sp. gr. Pn 3-bar has been confirmed. The structure refinement, Raman spectroscopy as well as magnetic measurements data of Bi{sub 3}FeSb{sub 2}O{sub 11} are presented. - Highlights: • The Bi{sub 2}O{sub 3}–Fe{sub 2}O{sub 3}–Sb{sub 2}O{sub 5} system phase diagram refinement has been performed. • The Bi{sub 3}FeSb{sub 2}O{sub 11} existence along with pyrochlore structure compound is shown. • It was determined that the Bi{sub 3}FeSb{sub 2}O{sub 11} is of disordered cubic KSbO{sub 3}-type structure. • Factor group

  19. Assessing the osteoblast transcriptome in a model of enhanced bone formation due to constitutive G{sub s}–G protein signaling in osteoblasts

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

    Wattanachanya, Lalita, E-mail: lalita_md@yahoo.com; Division of Endocrinology and Metabolism, Department of Medicine, Faculty of Medicine, Chulalongkorn University and King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok; Wang, Liping, E-mail: lipingwang05@yahoo.com

    G protein-coupled receptor (GPCR) signaling in osteoblasts (OBs) is an important regulator of bone formation. We previously described a mouse model expressing Rs1, an engineered constitutively active G{sub s}-coupled GPCR, under the control of the 2.3 kb Col I promoter. These mice showed a dramatic age-dependent increase in trabecular bone of femurs. Here, we further evaluated the effects of enhanced G{sub s} signaling in OBs on intramembranous bone formation by examining calvariae of 1- and 9-week-old Col1(2.3)/Rs1 mice and characterized the in vivo gene expression specifically occurring in osteoblasts with activated G{sub s} G protein-coupled receptor signaling, at the cellularmore » level rather than in a whole bone. Rs1 calvariae displayed a dramatic increase in bone volume with partial loss of cortical structure. By immunohistochemistry, Osterix was detected in cells throughout the inter-trabecular space while Osteocalcin was expressed predominantly in cells along bone surfaces, suggesting the role of paracrine mediators secreted from OBs driven by 2.3 kb Col I promoter could influence early OB commitment, differentiation, and/or proliferation. Gene expression analysis of calvarial OBs revealed that genes affected by Rs1 signaling include those encoding proteins important for cell differentiation, cytokines and growth factors, angiogenesis, coagulation, and energy metabolism. The set of G{sub s}-GPCRs and other GPCRs that may contribute to the observed skeletal phenotype and candidate paracrine mediators of the effect of G{sub s} signaling in OBs were also determined. Our results identify novel detailed in vivo cellular changes of the anabolic response of the skeleton to G{sub s} signaling in mature OBs. - Highlights: • OB expression of an engineered G{sub s}-coupled receptor dramatically increases bone mass. • We investigated the changes in gene expression in vivo in enhanced OB G{sub s} signaling. • Genes in cell cycle and transcription were

  20. Synthesis, crystal structure, and physical properties of the Gd{sub 3}BiO{sub 3} and Gd{sub 8}Bi{sub 3}O{sub 8} phases

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

    Forbes, Scott; Yuan, Fang; Kosuda, Kosuke

    The second and third known rare-earth bismuthide oxides, Gd{sub 3}BiO{sub 3} and Gd{sub 8}Bi{sub 3}O{sub 8}, have been discovered via high temperature reactions at 1300 °C. Like its Gd–Sb–O counterparts, the Gd{sub 3}BiO{sub 3} and Gd{sub 8}Bi{sub 3}O{sub 8} phases crystallize in the monoclinic C2/m space group, with the latter containing disordered Bi atoms along the b direction of the unit cell. Unlike the RE{sub 8}Sb{sub 3}O{sub 8} series, the formation of the Gd{sub 3}BiO{sub 3} phase does not necessarily precede the formation of Gd{sub 8}Bi{sub 3}O{sub 8}, which is likely due to the difficulty of accommodating bismuth in themore » RE–O framework due to its larger size. Physical property measurements performed on a pure Gd{sub 8}Bi{sub 3}O{sub 8} sample reveal semiconducting behavior. Although electronic structure calculations predict metallic behavior due to an unbalanced electron count, the semiconducting behavior originates from the Anderson localization of the Bi p states near the Fermi level as a result of atomic disorder. - Graphical abstract: Reaction of GdBi and Gd{sub 2}O{sub 3} at high temperatures yields Gd–Bi–O phases. - Highlights: • Gd{sub 3}BiO{sub 3} and Gd{sub 8}Bi{sub 3}O{sub 8}, the second and third rare-earth bismuthide oxides, have been discovered. • Gd{sub 3}BiO{sub 3} and Gd{sub 8}Bi{sub 3}O{sub 8} are isostructural with RE{sub 3}SbO{sub 3} and RE{sub 8}Sb{sub 3}O{sub 8}. • Gd{sub 8}Bi{sub 3}O{sub 8} displays semiconducting behavior despite an unbalanced electron count. • Anderson localization of Bi p states results in semiconducting behavior in Gd{sub 8}Bi{sub 3}O{sub 8}.« less

  1. In brown adipocytes, adrenergically induced β{sub 1}-/β{sub 3}-(G{sub s})-, α{sub 2}-(G{sub i})- and α{sub 1}-(G{sub q})-signalling to Erk1/2 activation is not mediated via EGF receptor transactivation

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

    Wang, Yanling; Fälting, Johanna M.; Mattsson, Charlotte L.

    2013-10-15

    Brown adipose tissue is unusual in that the neurotransmitter norepinephrine influences cell destiny in ways generally associated with effects of classical growth factors: regulation of cell proliferation, of apoptosis, and progression of differentiation. The norepinephrine effects are mediated through G-protein-coupled receptors; further mediation of such stimulation to e.g. Erk1/2 activation is in cell biology in general accepted to occur through transactivation of the EGF receptor (by external or internal pathways). We have examined here the significance of such transactivation in brown adipocytes. Stimulation of mature brown adipocytes with cirazoline (α{sub 1}-adrenoceptor coupled via G{sub q}), clonidine (α{sub 2} via G{submore » i}) or CL316243 (β{sub 3} via G{sub s}) or via β{sub 1}-receptors significantly activated Erk1/2. Pretreatment with the EGF receptor kinase inhibitor AG1478 had, remarkably, no significant effect on Erk1/2 activation induced by any of these adrenergic agonists (although it fully abolished EGF-induced Erk1/2 activation), demonstrating absence of EGF receptor-mediated transactivation. Results with brown preadipocytes (cells in more proliferative states) were not qualitatively different. Joint stimulation of all adrenoceptors with norepinephrine did not result in synergism on Erk1/2 activation. AG1478 action on EGF-stimulated Erk1/2 phosphorylation showed a sharp concentration–response relationship (IC{sub 50} 0.3 µM); a minor apparent effect of AG1478 on norepinephrine-stimulated Erk1/2 phosphorylation showed nonspecific kinetics, implying caution in interpretation of partial effects of AG1478 as reported in other systems. Transactivation of the EGF receptor is clearly not a universal prerequisite for coupling of G-protein coupled receptors to Erk1/2 signalling cascades. - Highlights: • In brown adipocytes, norepinephrine regulates proliferation, apoptosis, differentiation. • EGF receptor transactivation is supposed to

  2. Cu{sub 2}Mn{sub 1-x}Co{sub x}SnS{sub 4}: Novel keesterite type solid solutions

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

    Lopez-Vergara, F., E-mail: fer_martina@u.uchile.cl; Galdamez, A., E-mail: agaldamez@uchile.cl; Manriquez, V.

    2013-02-15

    A new family of Cu{sub 2}Mn{sub 1-x}Co{sub x}SnS{sub 4} chalcogenides has been synthesized by conventional solid-state reactions at 850 Degree-Sign C. The reactions products were characterized by powder X-ray diffraction (XRD), energy-dispersive X-ray analysis (SEM-EDS), Raman spectroscopy and magnetic susceptibility. The crystal structures of two members of the solid solution series Cu{sub 2}Mn{sub 0.4}Co{sub 0.6}SnS{sub 4} and Cu{sub 2}Mn{sub 0.2}Co{sub 0.8}SnS{sub 4} have been determined by single-crystal X-ray diffraction. Both phases crystallize in the tetragonal keesterite-type structure (space group I4{sup Macron }). The distortions of the tetrahedral volume of Cu{sub 2}Mn{sub 0.4}Co{sub 0.6}SnS{sub 4} and Cu{sub 2}Mn{sub 0.2}Co{sub 0.8}SnS{sub 4}more » were calculated and compared with the corresponding differences in the Cu{sub 2}MnSnS{sub 4} (stannite-type) end-member. The compounds show nearly the same Raman spectral features. Temperature-dependent magnetization measurements (ZFC/FC) and high-temperature susceptibility indicate that these solid solutions are antiferromagnetic. - Graphical abstract: View along [100] of the Cu{sub 2}Mn{sub 1-x}Co{sub x}SnS{sub 4} structure showing tetrahedral units and magnetic measurement ZFC-FC at 500 Oe. The insert shows the 1/{chi}-versus-temperature plot fitted by a Curie-Weiss law. Highlights: Black-Right-Pointing-Pointer Cu{sub 2}Mn{sub 1-x}Co{sub x}SnS{sub 4} solid solutions belong to the family of compounds adamantine. Black-Right-Pointing-Pointer Resolved single crystals of the solid solutions have space group I4{sup Macron }. Black-Right-Pointing-Pointer The distortion of the tetrahedral volume of Cu{sub 2}Mn{sub 1-x}Co{sub x}SnS{sub 4} were calculated. Black-Right-Pointing-Pointer These solid solutions are antiferromagnetic.« less

  3. Role of the Polymerase ϵ sub-unit DPB2 in DNA replication, cell cycle regulation and DNA damage response in Arabidopsis.

    PubMed

    Pedroza-Garcia, José Antonio; Domenichini, Séverine; Mazubert, Christelle; Bourge, Mickael; White, Charles; Hudik, Elodie; Bounon, Rémi; Tariq, Zakia; Delannoy, Etienne; Del Olmo, Ivan; Piñeiro, Manuel; Jarillo, Jose Antonio; Bergounioux, Catherine; Benhamed, Moussa; Raynaud, Cécile

    2016-09-06

    Faithful DNA replication maintains genome stability in dividing cells and from one generation to the next. This is particularly important in plants because the whole plant body and reproductive cells originate from meristematic cells that retain their proliferative capacity throughout the life cycle of the organism. DNA replication involves large sets of proteins whose activity is strictly regulated, and is tightly linked to the DNA damage response to detect and respond to replication errors or defects. Central to this interconnection is the replicative polymerase DNA Polymerase ϵ (Pol ϵ) which participates in DNA replication per se, as well as replication stress response in animals and in yeast. Surprisingly, its function has to date been little explored in plants, and notably its relationship with DNA Damage Response (DDR) has not been investigated. Here, we have studied the role of the largest regulatory sub-unit of Arabidopsis DNA Pol ϵ: DPB2, using an over-expression strategy. We demonstrate that excess accumulation of the protein impairs DNA replication and causes endogenous DNA stress. Furthermore, we show that Pol ϵ dysfunction has contrasting outcomes in vegetative and reproductive cells and leads to the activation of distinct DDR pathways in the two cell types. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  4. Treatment of adenoviral keratoconjunctivitis with a combination of povidone-iodine 1.0% and dexamethasone 0.1% drops: a clinical prospective controlled randomized study.

    PubMed

    Kovalyuk, Natalya; Kaiserman, Igor; Mimouni, Michael; Cohen, Ornit; Levartovsky, Shmuel; Sherbany, Hilda; Mandelboim, Michal

    2017-12-01

    To determine the efficacy of combination povidone-iodine (PVP-I) 1.0% eyedrops and dexamethasone 0.1% eyedrops in the treatment of adenoviral keratoconjunctivitis. In a prospective, randomized, controlled, double-blinded clinical trial patients with recent adenoviral keratoconjunctivitis (diagnosed clinically and confirmed by PCR), we randomly divided into three treatment groups: study group - received PVP-I 1.0% and dexamethasone 0.1%, control 1 group - received dexamethasone 0.1% and control 2 group - received lubricating eyedrops (hypromellose 0.3%). The treatment was administered four times a day in each group. All patients were examined and filled a questionnaire before treatment and on the 3rd, 5th and 7th days of treatment. We included in the study 78 eyes (26 in each group). Adenovirus type 8 was the most common pathogen (83% of cases). The fastest improvement in patients red eyes, discharge, superficial punctate keratitis and pseudomembranes was observed in the study group (p < 0.001). Those patients reached a near complete recovery in 5-7 days, which was also confirmed by reduction in Adenovirus titres by PCR. The slowest improvement was in the control 2 group. Subepithelial infiltrates (SEI) were observed in 44% of the control 1 group, 20% of the control 2 group and in 0% of the study group. The rate of reduction in Adenovirus titres was the slowest in the control 1 group. The combination of PVP-I 1.0% and dexamethasone 0.1% four times a day can reduce symptoms and expedite recovery in epidemic keratoconjunctivitis patients. © 2017 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

  5. CD11c identifies a subset of murine liver natural killer cells that responds to adenoviral hepatitis

    PubMed Central

    Burt, Bryan M.; Plitas, George; Stableford, Jennifer A.; Nguyen, Hoang M.; Bamboat, Zubin M.; Pillarisetty, Venu G.; DeMatteo, Ronald P.

    2008-01-01

    The liver contains a unique repertoire of immune cells and a particular abundance of NK cells. We have found that CD11c defines a distinct subset of NK cells (NK1.1+CD3−) in the murine liver whose function was currently unknown. In naïve animals, CD11c+ liver NK cells displayed an activated phenotype and possessed enhanced effector functions when compared with CD11c− liver NK cells. During the innate response to adenovirus infection, CD11c+ NK cells were the more common IFN-γ-producing NK cells in the liver, demonstrated enhanced lytic capability, and gained a modest degree of APC function. The mechanism of IFN-γ production in vivo depended on TLR9 ligation as well as IL-12 and -18. Taken together, our findings demonstrate that CD11c+ NK cells are a unique subset of NK cells in the murine liver that contribute to the defense against adenoviral hepatitis. PMID:18664530

  6. Formation of protein sub-visible particles during vacuum degassing of etanercept solutions.

    PubMed

    Wang, Haibin; Zheng, Hong-Jian; Wang, Zhao; Bai, Hua; Carpenter, John F; Chen, Shuqing; Fang, Wei-Jie

    2014-05-01

    The main purpose of this manuscript is to describe a phenomenon in which vacuum degassing a reconstituted freeze-dried fusion protein etanercept formulation caused a significant amount of protein sub-visible particles (SbVP). Physical stability of etanercept was monitored by micro-flow imaging (MFI), dynamic light scattering (DLS), size-exclusion high pressure liquid chromatography (SE-HPLC) and far- and near-ultraviolet circular dichroism (far- and near-UV CD). One potential explanation of this phenomenon is that bubble collapses when the vacuum is applied, leads to substantial heat formation, and ultimately free radical formation. Subsequently, the effect of a free-radical scavenger (ascorbic acid, AA) on SbVP formation was also evaluated. Degassing of etanercept solution by applying vacuum caused substantial increase of SbVP, as detected by MFI and DLS. However, traditional techniques such as SE-HPLC could not detect any change. The addition of free-radical scavenger had minimal effect on SbVP formation, therefore the formation of free radicals was probably not the main cause for this effect. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Crystallization and preliminary crystallographic analysis of the transpeptidase domain of penicillin-binding protein 2B from Streptococcus pneumoniae

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

    Yamada, Mototsugu, E-mail: mototsugu-yamada@meiji.co.jp; Watanabe, Takashi; Baba, Nobuyoshi

    The selenomethionyl-substituted transpeptidase domain of penicillin-binding protein (PBP) 2B from S. pneumoniae was isolated from a limited proteolysis digest of the soluble form of recombinant PBP 2B and then crystallized. MAD data were collected to 2.4 Å resolution. Penicillin-binding protein (PBP) 2B from Streptococcus pneumoniae catalyzes the cross-linking of peptidoglycan precursors that occurs during bacterial cell-wall biosynthesis. A selenomethionyl (SeMet) substituted PBP 2B transpeptidase domain was isolated from a limited proteolysis digest of a soluble form of recombinant PBP 2B and then crystallized. The crystals belonged to space group P4{sub 3}2{sub 1}2, with unit-cell parameters a = b = 86.39,more » c = 143.27 Å. Diffraction data were collected to 2.4 Å resolution using the BL32B2 beamline at SPring-8. The asymmetric unit contains one protein molecule and 63.7% solvent.« less

  8. Crystallization and preliminary X-ray diffraction studies of choline-binding protein F from Streptococcus pneumoniae

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

    Molina, Rafael; González, Ana; Moscoso, Miriam

    2007-09-01

    The modular choline-binding protein F (CbpF) from S. pneumoniae has been crystallized by the hanging-drop vapour-diffusion method. A SAD data set from a gadolinium-complex derivative has been collected to 2.1 Å resolution. Choline-binding protein F (CbpF) is a modular protein that is bound to the pneumococcal cell wall through noncovalent interactions with choline moieties of the bacterial teichoic and lipoteichoic acids. Despite being one of the more abundant proteins on the surface, along with the murein hydrolases LytA, LytB, LytC and Pce, its function is still unknown. CbpF has been crystallized using the hanging-drop vapour-diffusion method at 291 K. Diffraction-qualitymore » orthorhombic crystals belong to space group P2{sub 1}2{sub 1}2, with unit-cell parameters a = 49.13, b = 114.94, c = 75.69 Å. A SAD data set from a Gd-HPDO3A-derivatized CbpF crystal was collected to 2.1 Å resolution at the gadolinium L{sub III} absorption edge using synchrotron radiation.« less

  9. Chrystal structure properties of Al-doped Li{sub 4}Ti{sub 5}O{sub 12} synthesized by solid state reaction method

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

    Sandi, Dianisa Khoirum, E-mail: dianisa875@gmail.com; Suryana, Risa, E-mail: rsuryana@staff.uns.ac.id; Priyono, Slamet, E-mail: slam013@lipi.go.id

    2016-02-08

    This research aim is to analyze the effect of Aluminum (Al) doping in the structural properties of Al-doped Li{sub 4}Ti{sub 5}O{sub 12} as anode in lithium ion battery. Al-doped Li{sub 4}Ti{sub 5}O{sub 12} powders were synthesized by solid state reaction method. LiOH.H{sub 2}O, TiO{sub 2}, and Al{sub 2}O{sub 3} were raw materials. These materials were milled for 15 h, calcined at temperature of 750{sup o}C and sintered at temperature of 800{sup o}C. Mole percentage of doping Al (x) was varied at x=0; x=0.025; and x =0.05. Al-doped Li{sub 4}Ti{sub 5}O{sub 12} powders were synthesized by solid state reaction method. X-ray diffractionmore » was employed to determine the structure of Li{sub 4}Ti{sub 5}O{sub 12}. The PDXL software was performed on the x-ray diffraction data to estimate the phase percentage, the lattice parameter, the unit cell volume, and the crystal density. Al-doped Li{sub 4}Ti{sub 5}O{sub 12} has cubic crystal structure. Al-doping at x=0 and x=0.025 does not change the phase as Li{sub 4}Ti{sub 5}O{sub 12} while at x=0.050 the phase changes to the LiTiAlO{sub 4}. The diffraction patterns show that the angle shifted to the right as the increase of x which indicated that Al substitute Ti site. Percentage of Li{sub 4}Ti{sub 5}O{sub 12} phase at x=0 and x=0.025 was 97.8% and 96.8%, respectively. However, the lattice parameters, the unit cell volume, and the crystal density does not change significantly at x=0; x=0.025; and x=0.050. Based on the percentage of Li{sub 4}Ti{sub 5}O{sub 12} phase, the Al-doped Li at x=0 and x=0.025 is promising as a lithium battery anode.« less

  10. Adenoviral Expression of a Bispecific VHH-Based Neutralizing Agent That Targets Protective Antigen Provides Prophylactic Protection from Anthrax in Mice.

    PubMed

    Moayeri, Mahtab; Tremblay, Jacqueline M; Debatis, Michelle; Dmitriev, Igor P; Kashentseva, Elena A; Yeh, Anthony J; Cheung, Gordon Y C; Curiel, David T; Leppla, Stephen; Shoemaker, Charles B

    2016-01-06

    Bacillus anthracis, the causative agent of anthrax, secretes three polypeptides, which form the bipartite lethal and edema toxins (LT and ET, respectively). The common component in these toxins, protective antigen (PA), is responsible for binding to cellular receptors and translocating the lethal factor (LF) and edema factor (EF) enzymatic moieties to the cytosol. Antibodies against PA protect against anthrax. We previously isolated toxin-neutralizing variable domains of camelid heavy-chain-only antibodies (VHHs) and demonstrated their in vivo efficacy. In this work, gene therapy with an adenoviral (Ad) vector (Ad/VNA2-PA) (VNA, VHH-based neutralizing agents) promoting the expression of a bispecific VHH-based neutralizing agent (VNA2-PA), consisting of two linked VHHs targeting different PA-neutralizing epitopes, was tested in two inbred mouse strains, BALB/cJ and C57BL/6J, and found to protect mice against anthrax toxin challenge and anthrax spore infection. Two weeks after a single treatment with Ad/VNA2-PA, serum VNA2-PA levels remained above 1 μg/ml, with some as high as 10 mg/ml. The levels were 10- to 100-fold higher and persisted longer in C57BL/6J than in BALB/cJ mice. Mice were challenged with a lethal dose of LT or spores at various times after Ad/VNA2-PA administration. The majority of BALB/cJ mice having serum VNA2-PA levels of >0.1 μg/ml survived LT challenge, and 9 of 10 C57BL/6J mice with serum levels of >1 μg/ml survived spore challenge. Our findings demonstrate the potential for genetic delivery of VNAs as an effective method for providing prophylactic protection from anthrax. We also extend prior findings of mouse strain-based differences in transgene expression and persistence by adenoviral vectors. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  11. Protein preparation and preliminary X-ray crystallographic analysis of a putative glucosamine 6-phosphate deaminase from Streptococcus mutants

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

    Hu, Guan-Jing; Li, Lan-Fen; Li, Dan

    2007-09-01

    A glucosamine 6-phosphate deaminase homologue from S. mutans was expressed, purified and crystallized. Diffraction data have been collected to 2.4 Å resolution. The SMU.636 protein from Streptococcus mutans is a putative glucosamine 6-phosphate deaminase with 233 residues. The smu.636 gene was PCR-amplified from S. mutans genomic DNA and cloned into the expression vector pET-28a(+). The resultant His-tagged fusion protein was expressed in Escherichia coli and purified to homogeneity in two steps. Crystals of the fusion protein were obtained by the hanging-drop vapour-diffusion method. The crystals diffracted to 2.4 Å resolution and belong to space group P2{sub 1}2{sub 1}2{sub 1}, withmore » unit-cell parameters a = 53.83, b = 82.13, c = 134.70 Å.« less

  12. UTa{sub 2}O(S{sub 2}){sub 3}Cl{sub 6}: A ribbon structure containing a heterobimetallic 5d-5f M{sub 3} cluster

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

    Wells, Daniel M.; Chan, George H.; Ellis, Donald E.

    2010-02-15

    A new solid-state compound containing a heterobimetallic cluster of U and Ta, UTa{sub 2}O(S{sub 2}){sub 3}Cl{sub 6}, has been synthesized and its structure has been characterized by single-crystal X-ray diffraction methods. UTa{sub 2}O(S{sub 2}){sub 3}Cl{sub 6} was synthesized from UCl{sub 4} and Ta{sub 1.2}S{sub 2} at 883 K. The O is believed to have originated in the Ta{sub 1.2}S{sub 2} reactant. The compound crystallizes in the space group P1-bar of the triclinic system. The structure comprises a UTa{sub 2} unit bridged by mu{sub 2}-S{sub 2} and mu{sub 3}-O groups. Each Ta atom bonds to two mu{sub 2}-S{sub 2}, the mu{submore » 3}-O, and two terminal Cl atoms. Each U atom bonds to two mu{sub 2}-S{sub 2}, the mu{sub 3}-O, and four Cl atoms. The Cl atoms bridge in pairs to neighboring U atoms to form a ribbon structure. The bond distances are normal and are consistent with formal oxidation states of +IV/+V/-II/-I/-I for U/Ta/O/S/Cl, respectively. The optical absorbance spectrum displays characteristic transition peaks near the absorption edge. Density functional theory was used to assign these peaks to transitions between S{sup 1-} valence-band states and empty U 5f-6d hybrid bands. Density-of-states analysis shows overlap between Ta 5d and U bands, consistent with metal-metal interactions. - The UTa2O(S2)3Cl6 cluster with completed coordination sphere around uranium« less

  13. Permissive role of the acidification caused by wheat aleurone layers upon. alpha. -amylase induction by GA sub 3

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

    Rodriguez-Campos, E.; Bernal-Lugo, I.; Hamabata, A.

    1989-04-01

    Wheat aleurone has the capacity of acidifying the incubation medium in 1 to 2 pH units. The {alpha}-amylase induction by GA{sub 3} in isolated wheat aleurone layers is strongly dependent on acidic pH of the medium (pH < 5). To examine possible mechanisms {sup 35}-Met incorporation into proteins and {alpha}-amylase, in the presence of GA{sub 3} and Ca{sup 2+} at pH, 4, 5 and 6 was studied. Although {sup 35}-Met uptake decreased markedly ({approx} 90%) at pH 4 in thepresence of GA{sub 3}, incorporation into total protein did not change significantly from other conditions. Auto-radiography of SDS-PAGE showed that mostmore » of the amino acid was in the {alpha}-amylase band, meaning that the effect of acidic pH is specific for GA{sub 3} actions on aleurone tissue. On the other hand, an increase of protonated GA{sub 3} diffusion could be ruled out. Also, there was not {alpha}-amylase inactivation at pH 6. These findings point out to the important physiological role of the acidification caused by the aleurone.« less

  14. Purification, partial characterization, crystallization and preliminary X-ray diffraction of a novel cardiotoxin-like basic protein from Naja naja atra (South Anhui) venom

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

    Rong, Hui; Li, Yan; Lou, Xiao-hua

    2007-02-01

    A novel cardiotoxin-like basic protein from Naja naja atra was crystallized and diffraction data were collected to 2.35 Å resolution. A novel cardiotoxin-like basic protein was isolated from the venom of the Chinese cobra (Naja naja atra) from the south of Anhui in China. The protein inhibits the expression of vascular endothelial growth factor and basic fibroblast growth factor in human lung cancer cell line H1299 and induces the haemolysis of rabbit erythrocytes under low-lecithin conditions. After a two-step chromatographic purification, the resultant 7 kDa protein was crystallized by the hanging-drop vapour-diffusion method at room temperature. A complete data setmore » was collected to 2.35 Å resolution using an in-house X-ray diffraction system. The crystal belongs to space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 43.2, c = 147.9 Å. There are two molecules in the crystallographic asymmetric unit.« less

  15. Accelerating large-scale protein structure alignments with graphics processing units

    PubMed Central

    2012-01-01

    Background Large-scale protein structure alignment, an indispensable tool to structural bioinformatics, poses a tremendous challenge on computational resources. To ensure structure alignment accuracy and efficiency, efforts have been made to parallelize traditional alignment algorithms in grid environments. However, these solutions are costly and of limited accessibility. Others trade alignment quality for speedup by using high-level characteristics of structure fragments for structure comparisons. Findings We present ppsAlign, a parallel protein structure Alignment framework designed and optimized to exploit the parallelism of Graphics Processing Units (GPUs). As a general-purpose GPU platform, ppsAlign could take many concurrent methods, such as TM-align and Fr-TM-align, into the parallelized algorithm design. We evaluated ppsAlign on an NVIDIA Tesla C2050 GPU card, and compared it with existing software solutions running on an AMD dual-core CPU. We observed a 36-fold speedup over TM-align, a 65-fold speedup over Fr-TM-align, and a 40-fold speedup over MAMMOTH. Conclusions ppsAlign is a high-performance protein structure alignment tool designed to tackle the computational complexity issues from protein structural data. The solution presented in this paper allows large-scale structure comparisons to be performed using massive parallel computing power of GPU. PMID:22357132

  16. Structure refinement of Ba{sub 0.5}Sr{sub 0.5}Co{sub 0.8}Fe{sub 0.2}O{sub 3-d} as cathode materials for intermediate temperature solid oxide fuel cells (IT-SOFC)

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

    Zakaria, Nurhamidah, E-mail: nurhamidahzakaria@yahoo.com; Idris, Mohd Sobri, E-mail: sobri@unimap.edu.my; Osman, Rozana A. M., E-mail: rozana@unimap.edu.my

    2016-07-19

    Ba{sub 0.5}Sr{sub 0.5}Co{sub 0.8}Fe{sub 0.2}O{sub 3-δ} was successfully prepared using modified solid-state synthesis routes. The lowest temperature to obtained single phase of Ba{sub 0.5}Sr{sub 0.5}Co{sub 0.8}Fe{sub 0.2}O{sub 3-δ} is about 900°C for 15 hours. Longer period of time are required compared to only 5 hours at 950°C as established in literatures. The X-ray Diffraction (XRD) data confirmed that Ba{sub 0.5}Sr{sub 0.5}Co{sub 0.8}Fe{sub 0.2}O{sub 3-δ} is formed a cubic perovskite with the space group of Pm-3m. The lattice parameters of Ba{sub 0.5}Sr{sub 0.5}Co{sub 0.8}Fe{sub 0.2}O{sub 3-δ} are a = 3.990 (1) Å and unit cell volume is V = 63.5 (1)more » Å{sup 3}. The Rietveld refinement of XRD data revealed that the crystal structure of Ba{sub 0.5}Sr{sub 0.5}Co{sub 0.8}Fe{sub 0.2}O{sub 3-δ} slightly changes as a function of temperature.« less

  17. Inhibition of Choroidal Neovascularization by Intravenous Injection of Adenoviral Vectors Expressing Secretable Endostatin

    PubMed Central

    Mori, Keisuke; Ando, Akira; Gehlbach, Peter; Nesbitt, David; Takahashi, Kyoichi; Goldsteen, Donna; Penn, Michael; Chen, Cheauyan T.; Mori, Keiko; Melia, Michele; Phipps, Sandrina; Moffat, Diana; Brazzell, Kim; Liau, Gene; Dixon, Katharine H.; Campochiaro, Peter A.

    2001-01-01

    Endostatin is a cleavage product of collagen XVIII that inhibits tumor angiogenesis and growth. Interferon α2a blocks tumor angiogenesis and causes regression of hemangiomas, but has no effect on choroidal neovascularization (CNV). Therefore, inhibitors of tumor angiogenesis do not necessarily inhibit ocular neovascularization. In this study, we used an intravenous injection of adenoviral vectors containing a sig-mEndo transgene consisting of murine immunoglobulin κ-chain leader sequence coupled to sequence coding for murine endostatin to investigate the effect of high serum levels of endostatin on CNV in mice. Mice injected with a construct in which sig-mEndo expression was driven by the Rous sarcoma virus promoter had moderately high serum levels of endostatin and significantly smaller CNV lesions at sites of laser-induced rupture of Bruch’s membrane than mice injected with null vector. Mice injected with a construct in which sig-mEndo was driven by the simian cytomegalovirus promoter had ∼10-fold higher endostatin serum levels and had nearly complete prevention of CNV. There was a strong inverse correlation between endostatin serum level and area of CNV. This study provides proof of principle that gene therapy to increase levels of endostatin can prevent the development of CNV and may provide a new treatment for the leading cause of severe loss of vision in patients with age-related macular degeneration. PMID:11438478

  18. Probing of protein localization and shuttling in mitochondrial microcompartments by FLIM with sub-diffraction resolution.

    PubMed

    Söhnel, Anna-Carina; Kohl, Wladislaw; Gregor, Ingo; Enderlein, Jörg; Rieger, Bettina; Busch, Karin B

    2016-08-01

    The cell is metabolically highly compartmentalized. Especially, mitochondria host many vital reactions in their different microcompartments. However, due to their small size, these microcompartments are not accessible by conventional microscopy. Here, we demonstrate that time-correlated single-photon counting (TCSPC) fluorescence lifetime-imaging microscopy (FLIM) classifies not only mitochondria, but different microcompartments inside mitochondria. Sensor proteins in the matrix had a different lifetime than probes at membrane proteins. Localization in the outer and inner mitochondrial membrane could be distinguished by significant differences in the lifetime. The method was sensitive enough to monitor shifts in protein location within mitochondrial microcompartments. Macromolecular crowding induced by changes in the protein content significantly affected the lifetime, while oxidizing conditions or physiological pH changes had only marginal effects. We suggest that FLIM is a versatile and completive method to monitor spatiotemporal events in mitochondria. The sensitivity in the time domain allows for gaining substantial information about sub-mitochondrial localization overcoming diffraction limitation. This article is part of a Special Issue entitled 'EBEC 2016: 19th European Bioenergetics Conference, Riva del Garda, Italy, July 2-6, 2016', edited by Prof. Paolo Bernardi. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Multiple bonds between transition metals and main-group elements. 73. Synthetic routes to rhenium(V) alkyl and rhenium(VII) alkylidyne complexes. X-ray crystal structures of (. eta. sup 5 -C sub 5 Me sub 5 )Re( double bond O)(CH sub 3 )(CH sub 2 C(CH sub 3 ) sub 3 ) and (. eta. sup 5 -C sub 5 Me sub 5 )(Br) sub 3 Re triple bond CC(CH sub 3 ) sub 3

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

    Herrmann, W.A.; Felixberger, J.K.; Anwander, R.

    1990-05-01

    Dialkyloxo({eta}{sup 5}pentamethylcyclopentadienyl)rhenium(V) complexes ({eta}{sup 5}-C{sub 5}Me{sub 5})Re({double bond}O)(CH{sub 3})R{prime}(R{prime} = C{sub 2}H{sub 5}, CH{sub 2}Si(CH{sub 3}){sub 3}, CH{sub 2}C(CH{sub 3}){sub 3}), 1c-e, have become accessible through alkylation of ({eta}{sup 5}-C{sub 5}Me{sub 5})Re({double bond}O)(Cl)(CH{sub 3}) (7) with R{prime}MgCl. 1c-e are the first rhenium complexes containing different alkyl ligands. The neopentyl derivative 1e (R{prime} = CH{sub 2}C(CH{sub 3}){sub 3}) crystallizes in the orthorhombic space group Pbca with a = 960.7 (2), b = 2.844.5 (4), c = 1,260.7 (2) pm, and Z = 8. The X-ray crystal structure was refined to R{sub W} = 3.9%. The chiral molecule shows a distorted tetrahedralmore » geometry around the rhenium center. The tribromide 3b has been structurally characterized. Brown crystals of 3b belong to space group P2{sub 1}/c with unit cell dimensions a = 1,311.5 (2), b = 723.0 (1), c = 1,901.6 (2) pm, {beta} = 92.68 (1){degree}, and Z = 4. The structure exhibits a four-legged piano stool geometry with no trans influence of the neopentylidyne ligand to the bromine atom.« less

  20. Inflammatory mediator mRNA expression by adenovirus E1A-transfected bronchial epithelial cells.

    PubMed

    Higashimoto, Yuji; Elliott, W Mark; Behzad, Ali R; Sedgwick, Edward G; Takei, Tatsuo; Hogg, James C; Hayashi, Shizu

    2002-07-15

    Lung tissue from patients with emphysema and airway obstruction carries excess adenoviral E1A DNA that is expressed as protein in airway surface epithelium and is associated with an increased inflammatory response. To examine mechanisms by which latent adenoviral infection might amplify the inflammatory process, we transfected primary human bronchial epithelial (HBE) cells from three separate patients undergoing lung resection so that they stably expressed adenovirus E1A. Lipopolysaccharide stimulation of the E1A-transfected HBE cells increased intercellular adhesion molecule-1 and interleukin-8 mRNA and protein expression compared with control cells from the same patient. It also induced greater intercellular adhesion molecule-1 promoter activity and greater nuclear factor-kappa B binding activity of nuclear extracts in E1A transfectants than controls. E1A-positive transfectants constitutively expressed transforming growth factor-beta 1 mRNA and protein, whereas this expression was either very low or not detected in control cells. We conclude that adenoviral E1A transfection transforms primary HBE cells and upregulates their production of mediators that are clinically relevant to the pathogenesis of chronic obstructive pulmonary disease.

  1. Expression, Purification, Crystallization of Two Major Envelope Proteins from White Spot Syndrome Virus

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

    Tang,X.; Hew, C.

    2007-01-01

    White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapor-diffusion method. Crystals of SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 Mmore » sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 {angstrom} resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 {angstrom}. SeMet-labelled VP28 crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 {angstrom}, and diffracts to 2.0 {angstrom} resolution.« less

  2. Crystal structure of a new polar borate Na{sub 2}Ce{sub 2}[BO{sub 2}(OH)][BO{sub 3}]{sub 2} · H{sub 2}O with isolated boron triangles

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

    Topnikova, A. P.; Belokoneva, E. L., E-mail: elbel@geol.msu.ru; Dimitrova, O. V.

    2016-11-15

    Crystals of a new polar borate Na{sub 2}Ce{sub 2}[BO{sub 2}(OH)][BO{sub 3}]{sub 2} · H{sub 2}O were prepared by hydrothermal synthesis. The crystals are orthorhombic, a = 7.2295(7) Å, b = 11.2523(8) Å, c = 5.1285(6) Å, Z = 2, sp. gr. C2mm (Amm2), R = 0.0253. The formula of the compound was derived from the structure determination. The Ce and Na atoms are coordinated by nine and six O atoms, respectively. The Ce position is split, and a small amount of Ce is incorporated into the Na1 site with the isomorphous substitution for Na. The anionic moieties exist as isolatedmore » BO{sub 3} and BO{sub 2}(OH) triangles. The planes of the BO{sub 2}(OH) triangles with mm2 symmetry are parallel to the ab plane. The planes of the BO{sub 3} triangles with m symmetry are perpendicular to the ab plane and are rotated in a diagonal way. The splitting of the Ce positions and the polar arrangement of the BO{sub 2}(OH) triangles, water molecules, and Na atoms are observed along the polar a axis. The new structure is most similar to the new borate NaCa{sub 4}[BO{sub 3}]{sub 3} (sp. gr. Ama2), in which triangles of one type are arranged in a polar fashion along the c axis. Weak nonlinear-optical properties of both polar borates are attributed to the quenching of the second-harmonic generation due to the mutually opposite orientation of two-thirds of B triangles in the unit cell.« less

  3. Synthesis, structure, and characterization of two new bismuth(III) selenite/tellurite nitrates: [(Bi{sub 3}O{sub 2})(SeO{sub 3}){sub 2}](NO{sub 3}) and [Bi(TeO{sub 3})](NO{sub 3})

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

    Meng, Chang-Yu; Wei, Ming-Fang; Geng, Lei, E-mail: lgeng.cn@gmail.com

    Two new bismuth(III) selenite/tellurite nitrates, [(Bi{sub 3}O{sub 2})(SeO{sub 3}){sub 2}](NO{sub 3}) and [Bi(TeO{sub 3})](NO{sub 3}), have been synthesized by conventional facile hydrothermal method at middle temperature 200 °C and characterized by single-crystal X-ray diffraction, powder diffraction, UV–vis–NIR optical absorption spectrum, infrared spectrum and thermal analylsis. Both [(Bi{sub 3}O{sub 2})(SeO{sub 3}){sub 2}](NO{sub 3}) and [Bi(TeO3)](NO3) crystallize in the monoclinic centronsymmetric space group P2{sub 1}/c with a=9.9403(4) Å, b=9.6857(4) Å, c=10.6864(5) Å, β=93.1150(10)° for [(Bi{sub 3}O{sub 2})(SeO{sub 3}){sub 2}](NO{sub 3}) and a=8.1489(3) Å, b=9.0663(4) Å, c=7.4729(3) Å, β=114.899(2)° for Bi(TeO3)(NO3), respectively. The two compounds, whose structures are composed of three different asymmetricmore » building units, exhibit two different types of structures. The structure of [(Bi{sub 3}O{sub 2})(SeO{sub 3}){sub 2}](NO{sub 3}) features a three-dimensional (3D) bismuth(III) selenite cationic tunnel structure [(Bi{sub 3}O{sub 2})(SeO{sub 3}){sub 2}] {sup 3}{sub ∞} with NO{sub 3}{sup −} anion group filling in the 1D tunnel along b axis. The structure of [Bi(TeO{sub 3})](NO{sub 3}) features 2D bismuth(III) tellurite [Bi(TeO{sub 3}){sub 2}]{sup 2}{sub ∞} layers separated by NO{sub 3}{sup −} anion groups. The results of optical diffuse-reflectance spectrum measurements and electronic structure calculations based on density functional theory methods show that the two compounds are wide band-gap semiconductors. - Graphical abstract: Two novel bismuth{sup III} selenite/tellurite nitrates [(Bi{sub 3}O{sub 2})(SeO{sub 3}){sub 2}](NO{sub 3}) with 3D tunnel structure and [Bi(TeO{sub 3})](NO{sub 3}) with 2D layer structure have been firstly synthesized and characterized. Display Omitted - Highlights: • Two novel bismuth{sup III} nitrates [(Bi{sub 3}O{sub 2})(SeO{sub 3}){sub 2}](NO{sub 3}) and [Bi(TeO{sub 3})](NO{sub 3}) were

  4. Protein Molecular Structures, Protein SubFractions, and Protein Availability Affected by Heat Processing: A Review

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

    Yu,P.

    2007-01-01

    The utilization and availability of protein depended on the types of protein and their specific susceptibility to enzymatic hydrolysis (inhibitory activities) in the gastrointestine and was highly associated with protein molecular structures. Studying internal protein structure and protein subfraction profiles leaded to an understanding of the components that make up a whole protein. An understanding of the molecular structure of the whole protein was often vital to understanding its digestive behavior and nutritive value in animals. In this review, recently obtained information on protein molecular structural effects of heat processing was reviewed, in relation to protein characteristics affecting digestive behaviormore » and nutrient utilization and availability. The emphasis of this review was on (1) using the newly advanced synchrotron technology (S-FTIR) as a novel approach to reveal protein molecular chemistry affected by heat processing within intact plant tissues; (2) revealing the effects of heat processing on the profile changes of protein subfractions associated with digestive behaviors and kinetics manipulated by heat processing; (3) prediction of the changes of protein availability and supply after heat processing, using the advanced DVE/OEB and NRC-2001 models, and (4) obtaining information on optimal processing conditions of protein as intestinal protein source to achieve target values for potential high net absorbable protein in the small intestine. The information described in this article may give better insight in the mechanisms involved and the intrinsic protein molecular structural changes occurring upon processing.« less

  5. Guinea pig adenovirus infection does not inhibit cochlear transfection with human adenoviral vectors in a model of hearing loss.

    PubMed

    Hankenson, F Claire; Wathen, Asheley B; Eaton, Kathryn A; Miyazawa, Toru; Swiderski, Donald L; Raphael, Yehoash

    2010-04-01

    Routine surveillance of guinea pigs maintained within a barrier facility detected guinea pig adenovirus (GPAdV) in sentinel animals. These guinea pigs served as models of induced hearing loss followed by regeneration of cochlear sensory (hair) cells through transdifferentiation of nonsensory cells by using human adenoviral (hAV) gene therapy. To determine whether natural GPAdV infection affected the ability of hAV vectors to transfect inner ear cells, adult male pigmented guinea pigs (n = 7) were enrolled in this study because of their prolonged exposure to GPAdV-seropositive conspecifics. Animals were deafened chemically (n = 2), received an hAV vector carrying the gene for green fluorescent protein (hAV-GFP) surgically without prior deafening (n = 2), or were deafened chemically with subsequent surgical inoculation of hAV-GFP (n = 3). Cochleae were evaluated by using fluorescence microscopy, and GFP expression in supporting cells indicated that the hAV-GFP vector was able to transfect inner ears in GPAdV-seropositive guinea pigs that had been chemically deafened. Animals had histologic evidence of interstitial pneumonia, attributable to prior infection with GPAdV. These findings confirmed that the described guinea pigs were less robust animal models with diminished utility for the overall studies. Serology tests confirmed that 5 of 7 animals (71%) were positive for antibodies against GPAdV at necropsy, approximately 7 mo after initial detection of sentinel infection. Control animals (n = 5) were confirmed to be seronegative for GPAdV with clinically normal pulmonary tissue. This study is the first to demonstrate that natural GPAdV infection does not negatively affect transfection with hAV vectors into guinea pig inner ear cells, despite the presence of other health complications attributed to the viral infection.

  6. Effect of Mn and Ti substitution on the reflection loss characteristic of Ba{sub 0.6}Sr{sub 0.4}Fe{sub 11-z}MnTi{sub z}O{sub 19} (z = 0, 1, 2 and 3)

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

    Gunanto, Y. E., E-mail: yohanes.gunanto@uph.edu; Cahyadi, L., E-mail: lina.cahyadi@uph.edu; Adi, W. Ari, E-mail: dwisnuaa@batan.go.id

    2016-04-19

    The synthesis and characterization of composition Ba{sub 0.6}Sr{sub 0.4}Fe{sub 11-z}MnTi{sub z}O{sub 19} (z = 0; 1; 2 and 3) compound by solid state reaction using mechanical milling have been performed. The raw materials were BaCO{sub 3}, SrCO{sub 3}, Fe{sub 2}O{sub 3}, MnCO{sub 3}, and TiO{sub 2}. The mixed powder was compacted and sintered at 1000°C for 5 hours. X-ray diffraction studies indicate expansion of hexagonal unit cell and compression of atomic density with substitution of Mn{sup 2+} and Ti{sup 4+} ions. Effect of substitution upon magnetic properties revealed that total magnetization, remanence, and coercivity changed with substitution due to preferentialmore » site occupancy of substituted Mn{sup 2+} and Ti{sup 4+} ions. Since the coercivity and total magnetization may be controlled by substitution while maintaining resistive properties, this material is useful for microwave absorber.« less

  7. Dielectric relaxation in 0-3 PVDF-Ba(Fe{sub 1/2}Nb{sub 1/2})O{sub 3} composites

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

    Chandra, K. P., E-mail: kpchandra23@gmail.com; Singh, Rajan; Kulkarni, A. R., E-mail: ajit2957@gmail.com

    2016-05-06

    (1-x)PVDF-xBa(Fe{sub 1/2}Nb{sub 1/2})O{sub 3} ceramic-polymer composites with x = 0.025, 0.05, 0.10, 0.15 were prepared using melt-mixing technique. The crystal symmetry, space group and unit cell dimensions were determined from the XRD data of Ba(Fe{sub 1/2}Nb{sub 1/2})O{sub 3} using FullProf software, whereas crystallite size and lattice strain were estimated using Williamson-Hall approach. The distribution of Ba(Fe{sub 1/2}Nb{sub 1/2})O{sub 3} particles in the PVDF matrix were examined on the cryo-fractured surfaces using a scanning electron microscope. Cole-Cole and pseudo Cole-Cole analysis suggested the dielectric relaxation in this system to be of non-Debye type. Filler concentration dependent real and imaginary parts ofmore » dielectric constant as well as ac conductivity data followed definite trends of exponential growth types of variation.« less

  8. Synthesis, structure and electrochemical properties of LiNaCo{sub 0.5}Fe{sub 0.5}PO{sub 4}F fluoride-phosphate

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

    Fedotov, Stanislav S.; Skoltech Center for Electrochemical Energy Storage, Skolkovo Institute of Science and Technology, 143026 Moscow; Kuzovchikov, Sergey M.

    2016-10-15

    LiNaCo{sub 0.5}Fe{sub 0.5}PO{sub 4}F fluoride-phosphate was synthesized via conventional solid-state and novel freeze-drying routes. The crystal structure was refined based on neutron powder diffraction (NPD) data and validated by electron diffraction (ED) and high-resolution transmission electron microscopy (HRTEM). The alkali ions are ordered in LiNaCo{sub 0.5}Fe{sub 0.5}PO{sub 4}F and the transition metals jointly occupy the same crystallographic sites. The oxidation state and oxygen coordination environment of the Fe atoms were verified by {sup 57}Fe Mössbauer spectroscopy. Electrochemical tests of the LiNaCo{sub 0.5}Fe{sub 0.5}PO{sub 4}F cathode material demonstrated a reversible activity of the Fe{sup 3+}/Fe{sup 2+} redox couple at the electrodemore » potential near 3.4 V and minor activity of the Co{sup 3+}/Co{sup 2+} redox couple over 5 V vs Li/Li{sup +}. The material exhibited the discharge capacity of more than 82% (theo.) regarding Fe{sup 3+}/Fe{sup 2+} in the 2.4÷4.6 V vs Li/Li{sup +} potential range. - Graphical abstract: The ball-polyhedral representation of the LiNaCo{sub 0.5}Fe{sub 0.5}PO{sub 4}F crystal structure. The MO{sub 4}F{sub 2} units are depicted as blue octahedra, PO{sub 4} units as orange tetrahedra, sodium atoms are designated as yellow (Na1), lithium – red and brown (Li2, Li3 resp.), fluorine – green, oxygen – violet spheres. - Highlights: • Freeze-drying method was successfully applied to the synthesis of LiNaCo{sub 0.5}Fe{sub 0.5}PO{sub 4}F. • The crystal structure of LiNaCo{sub 0.5}Fe{sub 0.5}PO{sub 4}F was refined based on NPD and validated by ED and HRTEM. • LiNaCo{sub 0.5}Fe{sub 0.5}PO{sub 4}F demonstrated a reversible Li de/intercalation in the 2.5÷4.6 V vs Li/Li{sup +} range.« less

  9. High-pressure behaviour of Cs{sub 2}V{sub 3}O{sub 8} fresnoite

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

    Grzechnik, Andrzej, E-mail: grzechnik@xtal.rwth-aachen.de; Yeon, Jeongho; Zur Loye, Hans-Conrad

    2016-06-15

    Crystal structure of Cs{sub 2}V{sub 3}O{sub 8} fresnoite (P4bm, Z=2) has been studied using single-crystal X-ray diffraction in a diamond anvil cell to 8.6 GPa at room temperature. Cs{sub 2}V{sub 3}O{sub 8} undergoes a reversible first-order phase transition at about 4 GPa associated with anomalies in the pressure dependencies of the lattice parameters and unit-cell volume but without any symmetry change. Both structures consist of layers of corner-sharing V{sup 5+}O{sub 4} tetrahedra and V{sup 4+}O{sub 5} tetragonal pyramids separated by the Cs{sup +} cations located between the layers. At low pressures, the compression has little effect on the polarity ofmore » the structure. Above 4 GPa, the pseudosymmetry with respect to the corresponding centrosymmetric space group P4/mbm abruptly increases. The effects of external pressure and of the A{sup +} cation substitution in the vanadate fresnoites A{sub 2}V{sub 3}O{sub 8} (A{sup +}: K{sup +}, Rb{sup +}, NH{sub 4}{sup +}, Cs{sup +}) are discussed. - Graphical abstract: Non-centrosymmetric Cs{sub 2}V{sub 3}O{sub 8} undergoes a reversible first-order phase transition at about 4 GPa associated with an abrupt change of the pseudosymmetry with respect to the centrosymmetric space group P4/mbm. Display Omitted - Highlights: • High-pressure behaviour of vanadate fresnoites depends on alkali metal cations. • The size of the alkali metal cation determines whether the high-pressure phase is centrosymmetric. • No incommensurate structures are observed upon compression.« less

  10. Quadratic general rotary unitized design for doping concentrations and up-conversion luminescence properties of Er{sup 3+}/Yb{sup 3+} co-doped NaLa(MoO{sub 4}){sub 2} phosphors

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

    Sun, Jiashi, E-mail: sunjs@dlmu.edu.cn; Shi, Linlin; Li, Shuwei

    Highlights: • NaLa(MoO4)2: Er3+/Yb3+ phosphor is synthesized by solid state method. • QGRUD is first applied to the codoping concentration option. • Optimized phosphor presents more stable UC emissions than the commercial phosphor. - Abstract: It is still a great challenge that designing proper codoping concentrations of rare earth ions for achieving intensest expected emission from the studied phosphor. In this work, the quadratic general rotary unitized design (QGRUD) was introduced into the codoping concentration option of NaLa(MoO{sub 4}){sub 2}: Er{sup 3+}/Yb{sup 3+} phosphor for upconversion (UC) applications, and the optimum doping concentrations of Er{sup 3+} and Yb{sup 3+} formore » achieving maximum UC luminescence intensity, which is close to commercial NaYF{sub 4}:Er{sup 3+}/Yb{sup 3+} phosphor, were obtained. The two-photon process was assigned to the green UC emissions in the optimized NaLa(MoO{sub 4}){sub 2}: Er{sup 3+}/Yb{sup 3+} phosphor. It was also demonstrated that the optimized phosphor presented more stable upconversion emissions than the commercial NaYF{sub 4}:Er{sup 3+}/Yb{sup 3+} phosphor.« less

  11. Effects of configurational disorder on the elastic properties of icosahedral boron-rich alloys based on B{sub 6}O, B{sub 13}C{sub 2}, and B{sub 4}C, and their mixing thermodynamics

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

    Ektarawong, A., E-mail: anekt@ifm.liu.se; Hultman, L.; Birch, J.

    The elastic properties of alloys between boron suboxide (B{sub 6}O) and boron carbide (B{sub 13}C{sub 2}), denoted by (B{sub 6}O){sub 1−x}(B{sub 13}C{sub 2}){sub x}, as well as boron carbide with variable carbon content, ranging from B{sub 13}C{sub 2} to B{sub 4}C are calculated from first-principles. Furthermore, the mixing thermodynamics of (B{sub 6}O){sub 1−x}(B{sub 13}C{sub 2}){sub x} is studied. A superatom-special quasirandom structure approach is used for modeling different atomic configurations, in which effects of configurational disorder between the carbide and suboxide structural units, as well as between boron and carbon atoms within the units, are taken into account. Elastic propertiesmore » calculations demonstrate that configurational disorder in B{sub 13}C{sub 2}, where a part of the C atoms in the CBC chains substitute for B atoms in the B{sub 12} icosahedra, drastically increase the Young’s and shear modulus, as compared to an atomically ordered state, B{sub 12}(CBC). These calculated elastic moduli of the disordered state are in excellent agreement with experiments. Configurational disorder between boron and carbon can also explain the experimentally observed almost constant elastic moduli of boron carbide as the carbon content is changed from B{sub 4}C to B{sub 13}C{sub 2}. The elastic moduli of the (B{sub 6}O){sub 1−x}(B{sub 13}C{sub 2}){sub x} system are also practically unchanged with composition if boron-carbon disorder is taken into account. By investigating the mixing thermodynamics of the alloys, in which the Gibbs free energy is determined within the mean-field approximation for the configurational entropy, we outline the pseudo-binary phase diagram of (B{sub 6}O){sub 1−x}(B{sub 13}C{sub 2}){sub x}. The phase diagram reveals the existence of a miscibility gap at all temperatures up to the melting point. Also, the coexistence of B{sub 6}O-rich as well as ordered or disordered B{sub 13}C{sub 2}-rich domains in the material

  12. Wheat grain quality under enhanced tropospheric CO{sub 2} and O{sub 3} concentrations

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

    Rudorff, B.F.T.; Mulchi, C.L.; Fenny, P.

    It is expected that the progressive increase of tropospheric trace gases such as CO{sub 2} and O{sub 3} will have a significant impact on agricultural production. The single and combined effects of CO{sub 2} enrichment and tropospheric O{sub 3} on grain quality characteristics in soft red winter wheat (Triticum aestivum L.) were examined in field studies using 3 m in diam. open-top chambers. Wheat cultivars {open_quotes}Massey{close_quotes} (1991) and {open_quotes}Saluda{close_quotes} (1992) were exposed to two CO{sub 2} concentrations (350 vs. 500 {mu}mol CO{sub 2} mol{sup {minus}1}; 12 h d{sup {minus}1}) in combination with two O{sub 3} regimes (charcoal-filtered air vs. ambientmore » air + 40 {plus_minus} 20 nmol O{sub 3} mol{sup {minus}1}, 7 h d{sup {minus}1}; Monday to Friday) from late March until maturity in June. Grain quality characteristics investigated included: test weight, milling and baking quality, flour yield, protein content, softness equivalent, alkaline water retention capacity, and cookie diameter. In general, exposure of plants to either elevated CO{sub 2} or weekly chronic O{sub 3} episodes caused only small changes in grain quality. Milling and baking quality score were not significantly changed in response to treatments in both years. Flour yield was increased by elevated CO{sub 2} but this increase was counteracted when elevated CO{sub 2} was combined with chronic O{sub 3} exposure. Flour protein contents were increased by enhanced O{sub 3} under elevated CO{sub 2}. Although the single effect of either CO{sub 2} enrichment or chronic O{sub 3} exposure had some impact o grain quality characteristics, it was noted that the combined effect of these gases was minor. It is likely that the concomitant increase of CO{sub 2} and O{sub 3} in the troposphere will have no significant impact on wheat grain quality. 25 refs., 1 fig., 2 tabs.« less

  13. Purification and crystallization of the ABC-type transport substrate-binding protein OppA from Thermoanaerobacter tengcongensis

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

    Gao, Jinlan; Li, Xiaolu; Tsinghua-Peking Joint Center for Life Sciences, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, People's Republic of China

    2012-06-22

    Highlights: Black-Right-Pointing-Pointer We truncated the signal peptide of OppA{sub TTE0054} to make it express in Escherichia coli as a soluble protein. Black-Right-Pointing-Pointer Crystals of OppA{sub TTE0054} were grown by sitting-drop vapor diffusion method. Black-Right-Pointing-Pointer The crystal of OppA{sub TTE0054} diffracted to 2.25 A. -- Abstract: Di- and oligopeptide- binding protein OppAs play important roles in solute and nutrient uptake, sporulation, biofilm formation, cell wall muropeptides recycling, peptide-dependent quorum-sensing responses, adherence to host cells, and a variety of other biological processes. Soluble OppA from Thermoanaerobacter tengcongensis was expressed in Escherichia coli. The protein was found to be >95% pure with SDS-PAGEmore » after a series of purification steps and the purity was further verified by mass spectrometry. The protein was crystallized using the sitting-drop vapour-diffusion method with PEG 400 as the precipitant. Crystal diffraction extended to 2.25 A. The crystal belonged to space group C222{sub 1}, with unit-cell parameters of a = 69.395, b = 199.572, c = 131.673 A, and {alpha} = {beta} = {gamma} = 90 Degree-Sign .« less

  14. Developmental patterning of sub-epidermal cells in the outer integument of Arabidopsis seeds

    PubMed Central

    Fiume, Elisa; Coen, Olivier; Xu, Wenjia; Lepiniec, Loïc

    2017-01-01

    The seed, the reproductive unit of angiosperms, is generally protected by the seed coat. The seed coat is made of one or two integuments, each comprising two epidermal cells layers and, in some cases, extra sub-epidermal cell layers. The thickness of the seed-coat affects several aspects of seed biology such as dormancy, germination and mortality. In Arabidopsis, the inner integument displays one or two sub-epidermal cell layers that originate from periclinal cell divisions of the innermost epidermal cell layer. By contrast, the outer integument was considered to be two-cell layered. Here, we show that sub-epidermal chalazal cells grow in between the epidermal outer integument cell layers to create an incomplete three-cell layered outer integument. We found that the MADS box transcription factor TRANSPARENT TESTA 16 represses growth of the chalaza and formation of sub-epidermal outer integument cells. Finally, we demonstrate that sub-epidermal cells of the outer and inner integument respond differently to the repressive mechanism mediated by FERTILIZATION INDEPENDENT SEED Polycomb group proteins and to fertilization signals. Our data suggest that integument cell origin rather than sub-epidermal cell position underlies different responses to fertilization. PMID:29141031

  15. Structural study of (N{sub 2}H{sub 5},H){sub 2.9}U{sub 1.1}Ce{sub 0.9}(C{sub 2}O{sub 4}){sub 5}·10H{sub 2}O from a conventional X-ray diffraction diagram obtained on a powder synthesized by a fast vortex process

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

    Brackx, E., E-mail: Emmanuelle.brackx@cea.fr; Laval, J.P.; Dugne, O.

    2015-01-15

    In the context of research on U/minor actinides for nuclear fuel reprocessing in the transmutation process, developments are first studied with surrogates containing uranium and lanthanides to facilitate testing. The tests consist of precipitating and calcining a hydrazinium uranium/cerium oxalate. The structure of this oxalate had not been previously determined, but was necessary to validate the physicochemical mechanisms involved. The present study, firstly demonstrates the structural similarity of the U/Ce oxalate phase (N{sub 2}H{sub 5},H){sub 2.9}U{sub 1.1}Ce{sub 0.9}(C{sub 2}O{sub 4}){sub 5}·10H{sub 2}O, synthesized using a vortex precipitator for continuous synthesis of actinide oxalates, with previously known oxalates, crystallizing in P6{submore » 3}/mmc symmetry, obtained by more classical methods. This fast precipitation process induces massive nucleation of fine powders. Their structural and microstructural determination confirms that the raw and dried phases belong to the same structural family as (NH{sub 4}){sub 2}U{sub 2}(C{sub 2}O{sub 4}){sub 5}·0.7H{sub 2}O whose structure was described by Chapelet-Arab in P6{sub 3}/mmc symmetry, using single crystal data. However, they present an extended disorder inside the tunnels of the structure, even after drying at 100 °C, between water and hydrazinium ions. This disorder is directly related to the fast vortex method. This structure determination can be used as a basis for further semi-quantitative analysis on the U/minor actinides products formed under various experimental conditions. - Highlights: • Uranium cerium oxalate precipitate characterization by X-ray powder diffraction. • Morphology characterization by SEM analysis. • Structure determination by unit cell Rietveld refinement.« less

  16. Synthesis and crystal structure of a novel pentaborate, Na{sub 3}ZnB{sub 5}O{sub 10}

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

    Chen Xuean; Li Ming; Chang Xinan

    A novel ternary borate, trisodium zinc pentaborate, Na{sub 3}ZnB{sub 5}O{sub 10}, has been prepared by solid-state reaction at temperature below 750deg. C. The single-crystal X-ray structural analysis showed that Na{sub 3}ZnB{sub 5}O{sub 10} crystallizes in the monoclinic space group P2{sub 1}/n with a=6.6725(7)A, b=18.1730(10)A, c=7.8656(9)A, {beta}=114.604(6){sup o}, Z=4. It represents a new structure type in which double ring [B{sub 5}O{sub 10}]{sup 5-} building units are bridged by ZnO{sub 4} tetrahedra through common O atoms to form a two-dimensional {sub {approx}}{sup 2}[ZnB{sub 5}O{sub 10}]{sup 3-}-layer that affords one-dimensional channels running parallel to the [101] direction. Symmetry-center related {sub {approx}}{sup 2}[ZnB{sub 5}O{submore » 10}]{sup 3-} layers are stacked along the b-axis, with the interlayer void spaces and intralayer open channels filled by Na{sup +} cations to balance charge. The IR spectrum further confirms the presence of both BO{sub 3} and BO{sub 4} groups and UV-vis diffuse reflectance spectrum shows a band gap of about 3.2eV.« less

  17. [The measurement of nursing workload in a sub-intensive unit with the Nine Equivalents of Nursing Manpower Scale].

    PubMed

    D'Orazio, Alessia; Dragonetti, Antonella; Finiguerra, Ivana; Simone, Paola

    2015-01-01

    The measurement of nursing workload in a sub-intensive unit with the Nine Equivalents of Nursing Manpower Scale. The need to maximize the nursing manpower to patients complexity requires a careful assessment of the nursing workload. To measure the nursing workload in a sub-intensive care unit and to assess the impact of patients isolated for multidrug resistant microorganisms (MDR) and with delirium, on the nursing workload. From december 1 2014 to march 31 2015 the nursing workload of patients admitted to a semi intensive untit of a Turin Hospital was measured with Nine Equivalents of Nursing Manpower (NEMS) original and modified, adding 1 point score for patients isolated and with delirium (Richmond Agitation Sedation Scale). Admission and discharge times, and the activities performed in and out of the unit were registered. Two-hundred-thirty patients were daily assessed and no differences were observed in mean NEMS scores with the original and modified scale: december 17.3 vs 18.5; January 19.4 vs 20.2; February 19.9 vs 20.6; March 19.5 vs 20.1). mean scores did not change across shifts although on average 8 days a month the scores exceeded 21, identifiyng an excess workload and a need of a 2:1 patient/nurse ratio. The maximum workload was concentrated between 12.00 and 18.00 pm. The NEMS scale allows to measure the nursing workload. Apparently patients isolated and with delirium did not significantly impact on the nursing workload.

  18. New high-pressure polymorph of In{sub 2}S{sub 3} with defect Th{sub 3}P{sub 4}-type structure

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

    Lai, Xiaojing; Zhu, Feng; Wu, Ye

    The high pressure behavior of β-In{sub 2}S{sub 3} (I4{sub 1}/amd and Z=16) has been studied by in situ synchrotron radiation X-ray diffraction combined with diamond anvil cell up to 71.7 GPa. Three pressure-induced phase transitions are evidenced at ∼6.6 GPa, ∼11.1 GPa at room temperature and 35.6 GPa after the high-temperature annealing using a portable laser heating system. The new polymorph of In{sub 2}S{sub 3} at 35.6 GPa is assigned to the denser cubic defect Th{sub 3}P{sub 4} structure (I4¯3d and Z=5.333), whose unit-cell parameters are a=7.557(1) Å and V=431.6(2) Å{sup 3}. The Th{sub 3}P{sub 4}-type phase can be stablemore » at least up to 71.7 GPa and cannot be preserved at ambient pressure. The pressure–volume relationship is well described by the second-order Birch–Murnaghan Equation of State, which yields B{sub 0}=63(3) GPa and B{sub 0}′=4 (fixed) for the β-In{sub 2}S{sub 3} phase and B{sub 0}=87(3) GPa and B{sub 0}′=4 (fixed) for the defect Th{sub 3}P{sub 4}-type phase respectively. - Graphical abstract: The structure and Rietveld refinement of new polymorph the defect Th{sub 3}P{sub 4}-type In{sub 2}S{sub 3}. This structure was observed at 35.6 GPa after laser heating by X-ray diffraction. Display Omitted - Highlights: Three pressure-induced phase transitions of β-In{sub 2}S{sub 3} were observed. β-In{sub 2}S{sub 3} was stable up to 6.6 GPa. The defect Th{sub 3}P{sub 4}-type In{sub 2}S{sub 3} was identified at 35.6 GPa after laser heating and was stable up to 71.7 GPa. Elastic properties of β-In{sub 2}S{sub 3} and Th{sub 3}P{sub 4}-type In{sub 2}S{sub 3} are well presented by Birch–Murnaghan EoS.« less

  19. Synthesis and morphology of Ba{sub 1-x}RE{sub 2x/3}Nb{sub 2}O{sub 6} nanocrystals with tungsten bronze structure in RE{sub 2}O{sub 3}-BaO-Nb{sub 2}O{sub 5}-B{sub 2}O{sub 3} glasses (RE: Sm, Eu, Gd, Dy, Er)

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

    Ida, H.; Shinozaki, K.; Honma, T.

    2012-12-15

    Ba{sub 1-x}RE{sub 2x/3}Nb{sub 2}O{sub 6} nanocrystals with a tetragonal tungsten bronze (TTB) structure are synthesized using a conventional glass crystallization technique in 2.3RE{sub 2}O{sub 3}-27.4BaO-34.3Nb{sub 2}O{sub 5}-36B{sub 2}O{sub 3} (mol%) (RE=Sm, Eu, Gd, Dy, and Er) glasses. One sharp crystallization peak is observed at {approx}670 Degree-Sign C in both powdered and bulk glasses, and the formation of Ba{sub 1-x}RE{sub 2x/3}Nb{sub 2}O{sub 6} nanocrystals with unit cell parameters of a{approx}1.24 nm and c{approx}0.39 nm was confirmed. It is found from high resolution transmission electron microscope observations that the morphology of Ba{sub 1-x}RE{sub 2x/3}Nb{sub 2}O{sub 6} nanocrystals is ellipsoidal. Their average particlemore » size is in the range of 15-60 nm and decreases with decreasing ionic radius of RE{sup 3+} being present in the precursor glasses. The optical transparent crystallized glass (bulk) shows the total photoluminescence (PL) quantum yield of 53% in the visible region of Eu{sup 3+} ions, suggesting a high potential of Ba{sub 1-x}RE{sub 2x/3}Nb{sub 2}O{sub 6} nanocrystals as PL materials. - Graphical abstract: This figure shows a TEM photograph for the heat-treated (667 Degree-Sign C, 3 h) sample of 2.3Dy{sub 2}O{sub 3}-27.4BaO-34.3Nb{sub 2}O{sub 5}-36B{sub 2}O{sub 3}. An ellipsoidal-shaped Ba{sub 1-x}Dy{sub 2x/3}Nb{sub 2}O{sub 6} nanocrystal with diameters of 17 and 28 nm is observed. The ellipsoidal morphology is a common feature in Ba{sub 1-x}RE{sub 2x/3}Nb{sub 2}O{sub 6} nanocrystals synthesized by the crystallization of 2.3RE{sub 2}O{sub 3}-27.4BaO-34.3Nb{sub 2}O{sub 5}-36B{sub 2}O{sub 3} glasses. Highlights: Black-Right-Pointing-Pointer Ba{sub 1-x}RE{sub 2x/3}Nb{sub 2}O{sub 6} nanocrystals with a tetragonal tungsten bronze structure are synthesized. Black-Right-Pointing-Pointer A glass crystallization technique was applied. Black-Right-Pointing-Pointer The morphology of Ba{sub 1-x}RE{sub 2x/3}Nb{sub 2}O{sub 6} nanocrystals

  20. Crystallization of Spätzle, a cystine-knot protein involved in embryonic development and innate immunity in Drosophila melanogaster

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

    Hoffmann, Anita; Neumann, Piotr; Schierhorn, Angelika

    2008-08-01

    Crystallization of the cystine-knot protein Spätzle occurred following serendipitous limited degradation of the pro-Spätzle propeptide during the crystallization experiment. The Spätzle protein is involved in both the definition of the dorsal–ventral axis during embryonic development and in the adult innate immune response. The disulfide-linked dimeric cystine-knot protein has been expressed as a proprotein in inclusion bodies in Escherichia coli and refolded in vitro by rapid dilution. Initial orthorhombic crystals that diffracted to 7 Å resolution were obtained after three months by the sitting-drop vapour-diffusion method. Optimization of the crystallization conditions resulted in orthorhombic crystals (space group P2{sub 1}2{sub 1}2{sub 1},more » with unit-cell parameters a = 53.0, b = 59.2, c = 62.5 Å) that diffracted to 2.8 Å resolution in-house. The small volume of the asymmetric unit indicated that it was not possible for the crystals to contain the complete pro-Spätzle dimer. Mass spectrometry, N-terminal sequencing and Western-blot analysis revealed that the crystals contained the C-terminal disulfide-linked cystine-knot dimer. Comparison of various crystallization experiments indicated that degradation of the N-terminal prodomain was dependent on the buffer conditions.« less

  1. Expression, crystallization and preliminary X-ray crystallographic analyses of two N-terminal acetyltransferase-related proteins from Thermoplasma acidophilum

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

    Han, Sang Hee; Ha, Jun Yong; Kim, Kyoung Hoon

    2006-11-01

    An N-terminal acetyltransferase ARD1 subunit-related protein (Ta0058) and an N-terminal acetyltransferase-related protein (Ta1140) from T. acidophilum were crystallized. X-ray diffraction data were collected to 2.17 and 2.40 Å, respectively. N-terminal acetylation is one of the most common protein modifications in eukaryotes, occurring in approximately 80–90% of cytosolic mammalian proteins and about 50% of yeast proteins. ARD1 (arrest-defective protein 1), together with NAT1 (N-acetyltransferase protein 1) and possibly NAT5, is responsible for the NatA activity in Saccharomyces cerevisiae. In mammals, ARD1 is involved in cell proliferation, neuronal development and cancer. Interestingly, it has been reported that mouse ARD1 (mARD1{sup 225}) mediatesmore » ∊-acetylation of hypoxia-inducible factor 1α (HIF-1α) and thereby enhances HIF-1α ubiquitination and degradation. Here, the preliminary X-ray crystallographic analyses of two N-terminal acetyltransferase-related proteins encoded by the Ta0058 and Ta1140 genes of Thermoplasma acidophilum are reported. The Ta0058 protein is related to an N-terminal acetyltransferase complex ARD1 subunit, while Ta1140 is a putative N-terminal acetyltransferase-related protein. Ta0058 shows 26% amino-acid sequence identity to both mARD1{sup 225} and human ARD1{sup 235}.The sequence identity between Ta0058 and Ta1140 is 28%. Ta0058 and Ta1140 were overexpressed in Escherichia coli fused with an N-terminal purification tag. Ta0058 was crystallized at 297 K using a reservoir solution consisting of 0.1 M sodium acetate pH 4.6, 8%(w/v) polyethylene glycol 4000 and 35%(v/v) glycerol. X-ray diffraction data were collected to 2.17 Å. The Ta0058 crystals belong to space group P4{sub 1} (or P4{sub 3}), with unit-cell parameters a = b = 49.334, c = 70.384 Å, α = β = γ = 90°. The asymmetric unit contains a monomer, giving a calculated crystal volume per protein weight (V{sub M}) of 2.13 Å{sup 3} Da{sup −1} and a solvent

  2. Synthesis, crystal structures and ionic conductivities of Bi{sub 14}P{sub 4}O{sub 31} and Bi{sub 50}V{sub 4}O{sub 85}. Two members of the series Bi{sub 18-4m}M{sub 4m}O{sub 27+4m} (M=P, V) related to the fluorite-type structure

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

    Mauvy, F.; Launay, J.C.; Darriet, J.

    2005-06-15

    The two hitherto unknown compounds Bi{sub 14}P{sub 4}O{sub 31} and Bi{sub 50}V{sub 4}O{sub 85} were prepared by the direct solid-state reaction of Bi{sub 2}O{sub 3} and (NH{sub 4})H{sub 2}PO{sub 4} or V{sub 2}O{sub 5}, respectively. Bi{sub 14}P{sub 4}O{sub 31} crystallizes in a C-centred monoclinic symmetry (C2/c space group) with the unit-cell parameters: a=19.2745(2)A, b=11.3698(1)A, c=52.4082(2)A and {beta}=93.63(1){sup o} (Z=16). The symmetry of Bi{sub 50}V{sub 4}O{sub 85} is also monoclinic (I2/m space group) with lattice parameters of a=11.8123(3)A, b=11.7425(2)A, c=16.5396(2)A and {beta}=90.14(1){sup o} (Z=2). Both structures correspond to a fluorite-type superstructure where the Bi and P or V atoms are orderedmore » in the framework. An idealized structural model is proposed where the structures result of the stacking of mixed atomic layers of composition [Bi{sub 14}M{sub 4}O{sub 31}] and [Bi{sub 18}O{sub 27}] respectively. This new family can be formulated Bi{sub 18-4m}M{sub 4m}O{sub 27+4m} with M=P, V and where the parameter m (0=sub 14}M{sub 4}O{sub 31}] layers to the total number of layers in the sequence. Bi{sub 14}P{sub 4}O{sub 31} corresponds to m=1 when Bi{sub 50}V{sub 8}O{sub 85} corresponds to m=1/3. In this last case, the structural sequence is simply one [Bi{sub 14}V{sub 4}O{sub 31}] layer to two [Bi{sub 18}O{sub 27}] layers. As predicted by the proposed structural building principle, Bi{sub 14}P{sub 4}O{sub 31} is not a good ionic conductor. The conductivity at 650 deg. C is 4 orders of magnitude lower from those found in Bi{sub 46}M{sub 8}O{sub 89} (M=P, V) (m=2/3) and Bi{sub 50}V{sub 4}O{sub 85} (m=1/3)« less

  3. Protein conformation and disease : pathological consequences of analogous mutations in homologous proteins.

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

    Stevens, F. J.; Pokkuluri, P. R.; Schiffer, M.

    2000-12-19

    The antibody light chain variable domain (V{sub L}){sup 1} and myelin protein zero (MPZ) are representatives of the functionally diverse immunoglobulin superfamily. The V{sub L} is a subunit of the antigen-binding component of antibodies, while MPZ is the major membrane-linked constituent of the myelin sheaths that coat peripheral nerves. Despite limited amino acid sequence homology, the conformations of the core structures of the two proteins are largely superimposable. Amino acid variations in V{sub L} account for various conformational disease outcomes, including amyloidosis. However, the specific amino acid changes in V{sub L} that are responsible for disease have been obscured bymore » multiple concurrent primary structure alterations. Recently, certain demyelination disorders have been linked to point mutations and single amino acid polymorphisms in MPZ. We demonstrate here that some pathogenic variations in MPZ correspond to changes suspected of determining amyloidosis in V{sub L}. This unanticipated observation suggests that studies of the biophysical origin of conformational disease in one member of a superfamily of homologous proteins may have implications throughout the superfamily. In some cases, findings may account for overt disease; in other cases, due to the natural repertoire of inherited polymorphisms, variations in a representative protein may predict subclinical impairment of homologous proteins.« less

  4. Discovering the hidden sub-network component in a ranked list of genes or proteins derived from genomic experiments

    PubMed Central

    García-Alonso, Luz; Alonso, Roberto; Vidal, Enrique; Amadoz, Alicia; de María, Alejandro; Minguez, Pablo; Medina, Ignacio; Dopazo, Joaquín

    2012-01-01

    Genomic experiments (e.g. differential gene expression, single-nucleotide polymorphism association) typically produce ranked list of genes. We present a simple but powerful approach which uses protein–protein interaction data to detect sub-networks within such ranked lists of genes or proteins. We performed an exhaustive study of network parameters that allowed us concluding that the average number of components and the average number of nodes per component are the parameters that best discriminate between real and random networks. A novel aspect that increases the efficiency of this strategy in finding sub-networks is that, in addition to direct connections, also connections mediated by intermediate nodes are considered to build up the sub-networks. The possibility of using of such intermediate nodes makes this approach more robust to noise. It also overcomes some limitations intrinsic to experimental designs based on differential expression, in which some nodes are invariant across conditions. The proposed approach can also be used for candidate disease-gene prioritization. Here, we demonstrate the usefulness of the approach by means of several case examples that include a differential expression analysis in Fanconi Anemia, a genome-wide association study of bipolar disorder and a genome-scale study of essentiality in cancer genes. An efficient and easy-to-use web interface (available at http://www.babelomics.org) based on HTML5 technologies is also provided to run the algorithm and represent the network. PMID:22844098

  5. Purification, identification and preliminary crystallographic studies of Pru du amandin, an allergenic protein from Prunus dulcis

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

    Gaur, Vineet; Sethi, Dhruv K.; Salunke, Dinakar M., E-mail: dinakar@nii.res.in

    The purification, identification, crystallization and preliminary crystallographic studies of an allergy-related protein, Pru du amandin, from P. dulcis nuts are reported. Food allergies appear to be one of the foremost causes of hypersensitivity reactions. Nut allergies account for most food allergies and are often permanent. The 360 kDa hexameric protein Pru du amandin, a known allergen, was purified from almonds (Prunus dulcis) by ammonium sulfate fractionation and ion-exchange chromatography. The protein was identified by a BLAST homology search against the nonredundant sequence database. Pru du amandin belongs to the 11S legumin family of seed storage proteins characterized by the presencemore » of a cupin motif. Crystals were obtained by the hanging-drop vapour-diffusion method. The crystals belong to space group P4{sub 1} (or P4{sub 3}), with unit-cell parameters a = b = 150.7, c = 164.9 Å.« less

  6. Effects of sub-lethal neurite outgrowth inhibitory concentrations of chlorpyrifos oxon on cytoskeletal proteins and acetylcholinesterase in differentiating N2a cells

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

    Flaskos, J., E-mail: flaskos@vet.auth.gr; Nikolaidis, E.; Harris, W.

    2011-11-15

    Previous work in our laboratory has shown that sub-lethal concentrations (1-10 {mu}M) of chlorpyrifos (CPF), diazinon (DZ) and diazinon oxon (DZO) inhibit the outgrowth of axon-like neurites in differentiating mouse N2a neuroblastoma cells concomitant with altered levels and/or phosphorylation state of axonal cytoskeleton and growth-associated proteins. The aim of the present work was to determine whether chlorpyrifos oxon (CPO) was capable of inhibiting N2a cell differentiation in a similar manner. Using experimental conditions similar to our previous work, sub-lethal concentrations (1-10 {mu}M) of CPO were found to inhibit N2a cell differentiation. However, unlike previous studies with DZ and DZO, theremore » was a high level of sustained inhibition of acetylcholinesterase (AChE) in CPO treated cells. Impairment of neurite outgrowth was also associated with reduced levels of growth associated protein-43 and neurofilament heavy chain (NFH), and the distribution of NFH in cells stained by indirect immunofluorescence was disrupted. However, in contrast to previous findings for DZO, the absolute level of phosphorylated NFH was unaffected by CPO exposure. Taken together, the findings suggest that sub-lethal concentrations of CPO inhibit axon outgrowth in differentiating N2a cells and that this effect involves reduced levels of two proteins that play key roles in axon outgrowth and maintenance. Although the inhibition of neurite outgrowth is unlikely to involve AChE inhibition directly, further work will help to determine whether the persistent inhibition of AChE by CPO can account for the different effects induced by CPO and DZO on the levels of total and phosphorylated NFH. -- Highlights: Black-Right-Pointing-Pointer Sub-lethal levels of chlorpyrifos oxon inhibit neurite outgrowth in N2a cells Black-Right-Pointing-Pointer Acetylcholinesterase exhibits sustained inhibition throughout exposure Black-Right-Pointing-Pointer The levels of neurofilament heavy chain and

  7. Preparation and structure of BiCrTeO{sub 6}: A new compound in Bi–Cr–Te–O system. Thermal expansion studies of Cr{sub 2}TeO{sub 6}, Bi{sub 2}TeO{sub 6} and BiCrTeO{sub 6}

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

    Vats, Bal Govind; Phatak, Rohan; Krishnan, K.

    Graphical abstract: A new compound BiCrTeO{sub 6} in the Bi–Cr–Te–O system was prepared by solid state route and characterized by X-ray diffraction method. The crystal structure of BiCrTeO{sub 6} shows that there is one distinct site for bismuth (Bi) atom (pink color), one chromium rich (Cr/Te = 68/32) (blue/green color), one tellurium rich (Te/Cr = 68/32) sites (green/blue color), and one distinct site for oxygen (O) atom (red color) in the unit cell. All cations in this structure show an octahedral coordination with oxygen atoms at the corners. The thermogram (TG) of the compound in air shows that it ismore » stable up to 1103 K and decomposes thereafter. The thermal expansion behaviour of BiCrTeO{sub 6} was studied using high temperature X-ray diffraction method from room temperature to 923 K under vacuum of 10{sup −8} atmosphere and showed positive thermal expansion with the average volume thermal expansion coefficients of 16.0 × 10{sup −6}/K. - Highlights: • A new compound BiCrTeO{sub 6} in Bi–Cr–Te–O system was prepared and characterized. • The crystal structure of BiCrTeO{sub 6} was determined by Rietveld refinement method. • The structure of BiCrTeO{sub 6} shows an octahedral coordination for all the metal ions. • The thermal expansion behavior of BiCrTeO{sub 6} from room temperature to 923 K showed a positive thermal expansion. • The average volume thermal expansion coefficient for BiCrTeO{sub 6} is 16.0 × 10{sup −6}/K. - Abstract: A new compound BiCrTeO{sub 6} in Bi–Cr–Te–O system was prepared by solid state reaction of Bi{sub 2}O{sub 3}, Cr{sub 2}O{sub 3} and H{sub 6}TeO{sub 6} in oxygen and characterized by X-ray diffraction (XRD) method. It could be indexed on a trigonal lattice, with the space group P-31c, unit cell parameters a = 5.16268(7) Å and c = 9.91861(17) Å. The crystal structure of BiCrTeO{sub 6} was determined by Rietveld refinement method using the powder XRD data. Structure shows that there is one

  8. Syntheses, crystal structures, and properties of new layered tungsten(VI)-containing materials based on the hexagonal-WO{sub 3} structure: M{sub 2}(WO{sub 3}){sub 3}SeO{sub 3} (M = NH{sub 4}, Rb, Cs)

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

    Harrison, W.T.A.; Dussack, L.L.; Jacobson, A.J.

    The hydrothermal syntheses and crystal structures of (NH{sub 4}){sub 2}(WO{sub 3}){sub 3}SeO{sub 3} and Cs{sub 2}(WO{sub 3}){sub 3}SeO{sub 3}, two new noncentrosymmetric, layered tungsten(VI)-containing phases are reported. Infrared, Raman, and thermogravimetric data are also presented. (NH{sub 4}){sub 2}(WO{sub 3}){sub 3}SeO{sub 3} and Cs{sub 2}(WO{sub 3}){sub 3}SeO{sub 3} are isostructural phases built up from hexagonal-tungsten-oxide-like, anionic layers of vertex-sharing WO{sub 6} octahedra, capped on one side by Se atoms (as selenite groups). Interlayer NH{sub 4}{sup +} or Cs{sup +} cations provide charge balance. The full H-bonding scheme in (NH{sub 4}){sub 2}(WO{sub 3}){sub 3}SeO{sub 3} has been elucidated from Rietveld refinement againstmore » neutron powder diffraction data. The WO{sub 6} octahedra display a 3 short + 3 long W-O bond-distance distribution within the WO{sub 6} unit in both these phases. (NH{sub 4}){sub 2}(WO{sub 3}){sub 3}SeO{sub 3} and Cs{sub 2}(WO{sub 3}){sub 3}SeO{sub 3} are isostructural with their molybdenum(VI)-containing analogues (NH{sub 4}){sub 2}(MoO{sub 3}){sub 3}SeO{sub 3} and Cs{sub 2} (MoO{sub 3}){sub 3}SeO{sub 3}. Crystal data: (NH{sub 4}){sub 2}(WO{sub 3}){sub 3}SeO{sub 3}, M{sub r} = 858.58, hexagonal, space group P6{sub 3} (No. 173), a = 7.2291(2) {angstrom}, c = 12.1486(3) {angstrom}, V = 549.82(3) {angstrom}{sup 3}, Z = 2, R{sub p} = 1.81%, and R{sub wp} = 2.29% (2938 neutron powder data). Cs{sub 2}(WO{sub 3}){sub 3}SeO{sub 3}, M{sub r} = 1088.31, hexagonal, space group P6{sub 3} (no. 173), a = 7.2615(2) {angstrom}, c = 12.5426(3) {angstrom}{sup 3}, Z = 2, R{sub p} = 4.84%, and R{sub wp} = 5.98% (2588 neutron powder data).« less

  9. Quantitative analysis of residual protein contamination of podiatry instruments reprocessed through local and central decontamination units

    PubMed Central

    2011-01-01

    Background The cleaning stage of the instrument decontamination process has come under increased scrutiny due to the increasing complexity of surgical instruments and the adverse affects of residual protein contamination on surgical instruments. Instruments used in the podiatry field have a complex surface topography and are exposed to a wide range of biological contamination. Currently, podiatry instruments are reprocessed locally within surgeries while national strategies are favouring a move toward reprocessing in central facilities. The aim of this study was to determine the efficacy of local and central reprocessing on podiatry instruments by measuring residual protein contamination of instruments reprocessed by both methods. Methods The residual protein of 189 instruments reprocessed centrally and 189 instruments reprocessed locally was determined using a fluorescent assay based on the reaction of proteins with o-phthaldialdehyde/sodium 2-mercaptoethanesulfonate. Results Residual protein was detected on 72% (n = 136) of instruments reprocessed centrally and 90% (n = 170) of instruments reprocessed locally. Significantly less protein (p < 0.001) was recovered from instruments reprocessed centrally (median 20.62 μg, range 0 - 5705 μg) than local reprocessing (median 111.9 μg, range 0 - 6344 μg). Conclusions Overall, the results show the superiority of central reprocessing for complex podiatry instruments when protein contamination is considered, though no significant difference was found in residual protein between local decontamination unit and central decontamination unit processes for Blacks files. Further research is needed to undertake qualitative identification of protein contamination to identify any cross contamination risks and a standard for acceptable residual protein contamination applicable to different instruments and specialities should be considered as a matter of urgency. PMID:21219613

  10. Quantitative analysis of residual protein contamination of podiatry instruments reprocessed through local and central decontamination units.

    PubMed

    Smith, Gordon Wg; Goldie, Frank; Long, Steven; Lappin, David F; Ramage, Gordon; Smith, Andrew J

    2011-01-10

    The cleaning stage of the instrument decontamination process has come under increased scrutiny due to the increasing complexity of surgical instruments and the adverse affects of residual protein contamination on surgical instruments. Instruments used in the podiatry field have a complex surface topography and are exposed to a wide range of biological contamination. Currently, podiatry instruments are reprocessed locally within surgeries while national strategies are favouring a move toward reprocessing in central facilities. The aim of this study was to determine the efficacy of local and central reprocessing on podiatry instruments by measuring residual protein contamination of instruments reprocessed by both methods. The residual protein of 189 instruments reprocessed centrally and 189 instruments reprocessed locally was determined using a fluorescent assay based on the reaction of proteins with o-phthaldialdehyde/sodium 2-mercaptoethanesulfonate. Residual protein was detected on 72% (n = 136) of instruments reprocessed centrally and 90% (n = 170) of instruments reprocessed locally. Significantly less protein (p < 0.001) was recovered from instruments reprocessed centrally (median 20.62 μg, range 0 - 5705 μg) than local reprocessing (median 111.9 μg, range 0 - 6344 μg). Overall, the results show the superiority of central reprocessing for complex podiatry instruments when protein contamination is considered, though no significant difference was found in residual protein between local decontamination unit and central decontamination unit processes for Blacks files. Further research is needed to undertake qualitative identification of protein contamination to identify any cross contamination risks and a standard for acceptable residual protein contamination applicable to different instruments and specialities should be considered as a matter of urgency.

  11. Inhibition of adenovirus multiplication by short interfering RNAs directly or indirectly targeting the viral DNA replication machinery.

    PubMed

    Kneidinger, Doris; Ibrišimović, Mirza; Lion, Thomas; Klein, Reinhard

    2012-06-01

    Human adenoviruses are a common threat to immunocompromised patients, e.g., HIV-positive individuals or solid-organ and, in particular, allogeneic stem cell transplant recipients. Antiviral drugs have a limited effect on adenoviruses, and existing treatment modalities often fail to prevent fatal outcome. Silencing of viral genes by short interfering RNAs (siRNAs) holds a great promise in the treatment of viral infections. The aim of the present study was to identify adenoviral candidate targets for RNA interference-mediated inhibition of adenoviral replication. We investigated the impact of silencing of a set of early, middle, and late viral genes on the replication of adenovirus 5 in vitro. Adenovirus replication was inhibited by siRNAs directed against the adenoviral E1A, DNA polymerase, preterminal protein (pTP), IVa2, hexon, and protease genes. Silencing of early and middle genes was more effective in inhibiting adenovirus multiplication than was silencing of late genes. A siRNA directed against the viral DNA polymerase mRNA decreased viral genome copy numbers and infectious virus progeny by several orders of magnitude. Since silencing of any of the early genes directly or indirectly affected viral DNA synthesis, our data suggest that reducing viral genome copy numbers is a more promising strategy for the treatment of adenoviral infections than is reducing the numbers of proteins necessary for capsid generation. Thus, adenoviral DNA replication was identified as a key target for RNAi-mediated inhibition of adenovirus multiplication. In addition, the E1A transcripts emerged as a second important target, because its knockdown markedly improved the viability of cells at late stages of infection. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. Protective Effect of High Molecular Weight Protein Sub-fraction of Calotropis procera Latex in Monoarthritic Rats.

    PubMed

    Chaudhary, Priyanka; Ramos, Marcio V; Vasconcelos, Mirele da Silveira; Kumar, Vijay L

    2016-05-01

    Proteins present in the latex of Calotropis procera have been shown to produce anti-inflammatory effect and to afford protection in various disease models. To determine the efficacy of high molecular weight protein sub-fraction (LPPI) of latex of C. procera in ameliorating joint inflammation and hyperalgesia in a preclinical model of arthritis. Monoarthritis was induced in rats by intra-articular injection of Freund's complete adjuvant (FCA) and the effect of two doses of LPPI (5 and 25 mg/kg) and diclofenac (5 mg/kg) was evaluated on joint swelling, stair climbing ability, motility, and dorsal flexion pain on day 3. The rats were sacrificed on day 3 to measure tissue levels of reduced glutathione (GSH) and thiobarbituric acid reactive substances (TBARS). Evaluation of joint histology was also made. Intra-articular injection of FCA produced joint swelling and difficulty in stair climbing ability, motility, and pain on flexion of the joint as revealed by scores obtained for these functional parameters. LPPI produced a dose-dependent decrease in joint swelling and improved joint functions. Arthritic rats also revealed altered oxidative homeostasis where joint tissue GSH levels were decreased and TBARS levels were increased as compared to normal rats. The levels of these oxidative stress markers were near normal in arthritic rats treated with LPPI. Moreover, treatment with LPPI also maintained the structural integrity of the joint. The protective effect of LPPI was comparable to the standard anti-inflammatory drug, diclofenac. The findings of the present study show that LPPI fraction comprising high molecular weight proteins could be used for the alleviation of arthritic symptoms. High molecular weight protein sub-fraction of latex of Calotropis procera (LPPI) reduced joint swelling and hyperalgesia in arthritic ratsLPPI produced a significant improvement in stair climbing ability and motility in arthritic ratsLPPI normalized the levels of oxidative stress markers in

  13. Na{sub 6}B{sub 13}O{sub 22.5}, a new noncentrosymmetric sodium borate

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

    Penin, N.; Touboul, M.; Nowogrocki, G.

    2005-03-15

    Na{sub 6}B{sub 13}O{sub 22.5} (B/Na=2.17) single crystals were obtained by heating, melting and appropriately cooling borax, Na{sub 2}[B{sub 4}O{sub 5}(OH){sub 4}].8H{sub 2}O. Its formula has been determined by the resolution of the structure from single-crystal X-ray diffraction data. The compound crystallizes in the noncentrosymmetric orthorhombic Iba2 space group, with the following unit cell parameters: a=33.359(11)A, b=9.554(3)A, c=10.644(4)A; V=3392.4(19)A{sup 3}; Z=8. The crystal structure was solved from 3226 reflections until R{sub 1}=0.0385. It exhibits a three-dimensional framework built up from BO{sub 3} triangles ({delta}) and BO{sub 4} tetrahedra (T). Two kinds of borate groups can be considered forming two different doublemore » B{sub 3}O{sub 3} rings: two B{sub 4}O{sub 9} (linkage by two boron atoms) and one B{sub 5}O{sub 11} (linkage by one boron atom); the shorthand notation of the new fundamental building block (FBB) existing in this compound is: 13: {infinity}{sup 3} [(5: 3{delta}+2T)+2(4: 2{delta}+2T)]. The discovery of this new borate questions the real number of Na{sub 2}B{sub 4}O{sub 7} varieties. The existence of Na{sub 6}B{sub 13}O{sub 22.5} (B/Na=2.17) and of another recently discovered borate, Na{sub 3}B{sub 7}O{sub 12} (B/Na=2.33; FBB 7: {infinity}{sup 3} [(3: 2{delta}+T)+(3: {delta}+2T)+(1: {delta})], with a composition close to the long-known borate {alpha}-Na{sub 2}B{sub 4}O{sub 7} (B/Na=2; FBB 8: {infinity}{sup 3} [(5: 3{delta}+2T)+(3: 2{delta}+T)], may explain the very complex equilibria reported in the Na{sub 2}O-B{sub 2}O{sub 3} phase diagram, especially in this range of composition.« less

  14. Persistent Socioeconomic and Political Dilemmas to the Implementation of the 1989 United Nations' Convention on the Rights of the Child in Sub-Saharan Africa

    ERIC Educational Resources Information Center

    Mulinge, Munyae M.

    2010-01-01

    Objective: The aim of this article is to revisit the subject of the implementation of the 1989 United Nations Convention on the Rights of the Child in Africa. Specifically, the article is an update to a previous article titled "Implementing the 1989 United Nations' Convention on the Rights of the Child in sub-Saharan Africa: The Overlooked…

  15. A Method for Visualization of Incoming Adenovirus Chromatin Complexes in Fixed and Living Cells

    PubMed Central

    Komatsu, Tetsuro; Dacheux, Denis; Kreppel, Florian; Nagata, Kyosuke; Wodrich, Harald

    2015-01-01

    Inside the adenovirus virion, the genome forms a chromatin-like structure with viral basic core proteins. Core protein VII is the major DNA binding protein and was shown to remain associated with viral genomes upon virus entry even after nuclear delivery. It has been suggested that protein VII plays a regulatory role in viral gene expression and is a functional component of viral chromatin complexes in host cells. As such, protein VII could be used as a maker to track adenoviral chromatin complexes in vivo. In this study, we characterize a new monoclonal antibody against protein VII that stains incoming viral chromatin complexes following nuclear import. Furthermore, we describe the development of a novel imaging system that uses Template Activating Factor-I (TAF-I/SET), a cellular chromatin protein tightly bound to protein VII upon infection. This setup allows us not only to rapidly visualize protein VII foci in fixed cells but also to monitor their movement in living cells. These powerful tools can provide novel insights into the spatio-temporal regulation of incoming adenoviral chromatin complexes. PMID:26332038

  16. RKIP phosphorylation–dependent ERK1 activation stimulates adipogenic lipid accumulation in 3T3-L1 preadipocytes overexpressing LC3

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

    Hahm, Jong Ryeal; Institute of Health Sciences, Gyeongsang National University School of Medicine, JinJu, 527-27; Ahmed, Mahmoud

    3T3-L1 preadipocytes undergo adipogenesis in response to treatment with dexamethaxone, 1-methyl-3-isobutylxanthine, and insulin (DMI) through activation of several adipogenic transcription factors. Many autophagy-related proteins are also highly activated in the earlier stages of adipogenesis, and the LC3 conjugation system is required for formation of lipid droplets. Here, we investigated the effect of overexpression of green fluorescent protein (GFP)-LC3 fusion protein on adipogenesis. Overexpression of GFP-LC3 in 3T3-L1 preadipocytes using poly-L-lysine-assisted adenoviral GFP-LC3 transduction was sufficient to produce intracellular lipid droplets. Indeed, GFP-LC3 overexpression stimulated expression of some adipogenic transcription factors (e.g., C/EBPα or β, PPARγ, SREBP2). In particular, SREBP2 wasmore » highly activated in preadipocytes transfected with adenoviral GFP-LC3. Also, phosphorylation of Raf kinase inhibitory protein (RKIP) at serine 153, consequently stimulating extracellular-signal regulated kinase (ERK)1 activity, was significantly increased during adipogenesis induced by either poly-L-lysine-assisted adenoviral GFP-LC3 transduction or culture in the presence of dexamethasone, 1-methyl-3-isobutylxanthine, and insulin. Furthermore, RKIP knockdown promoted ERK1 and PPARγ activation, and significantly increased the intracellular accumulation of triacylglycerides in DMI-induced adipogenesis. In conclusion, GFP-LC3 overexpression in 3T3-L1 preadipocytes stimulates adipocyte differentiation via direct modulation of RKIP-dependent ERK1 activity. - Highlights: • Overexpression of GFP-LC3 in 3T3-L1 cells produces intracellular lipid droplets. • SREBP2 is highly activated in preadipocytes transfected with adenoviral GFP-LC3. • RKIP phosphorylation at serine 153 is significantly increased during adipogenesis. • RKIP knockdown promotes ERK1 and PPARγ activation during adipogenesis. • RKIP-dependent ERK1 activation increases

  17. Imaging analysis of nuclear antiviral factors through direct detection of incoming adenovirus genome complexes.

    PubMed

    Komatsu, Tetsuro; Will, Hans; Nagata, Kyosuke; Wodrich, Harald

    2016-04-22

    Recent studies involving several viral systems have highlighted the importance of cellular intrinsic defense mechanisms through nuclear antiviral proteins that restrict viral propagation. These factors include among others components of PML nuclear bodies, the nuclear DNA sensor IFI16, and a potential restriction factor PHF13/SPOC1. For several nuclear replicating DNA viruses, it was shown that these factors sense and target viral genomes immediately upon nuclear import. In contrast to the anticipated view, we recently found that incoming adenoviral genomes are not targeted by PML nuclear bodies. Here we further explored cellular responses against adenoviral infection by focusing on specific conditions as well as additional nuclear antiviral factors. In line with our previous findings, we show that neither interferon treatment nor the use of specific isoforms of PML nuclear body components results in co-localization between incoming adenoviral genomes and the subnuclear domains. Furthermore, our imaging analyses indicated that neither IFI16 nor PHF13/SPOC1 are likely to target incoming adenoviral genomes. Thus our findings suggest that incoming adenoviral genomes may be able to escape from a large repertoire of nuclear antiviral mechanisms, providing a rationale for the efficient initiation of lytic replication cycle. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. GABA/sub B/ receptor activation inhibits Ca/sup 2 +/-activated potassium channels in synaptosomes: involvement of G-proteins

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

    Ticku, M.K.; Delgado, A.

    1989-01-01

    /sup 86/Rb-efflux assay from preloaded synaptosomes of rat cerebral cortex was developed to study the effect of GABA/sub B/ receptor agonist baclofen on Ca/sup 2 +/-activated K/sup +/-channels. Depolarization of /sup 86/Rb-loaded synaptosomes in physiological buffer increased Ca/sup 2 +/-activated /sup 86/Rb-efflux by 400%. The /sup 86/Rb-efflux was blocked by quinine sulfate, tetraethylammonium, and La/sup 3 +/ indicating the involvement of Ca/sup 2 +/-activated K/sup +/-channels. (-)Baclofen inhibited Ca/sup 2 +/-activated /sup 86/Rb-efflux in a stereospecific manner. The inhibitory effect of (-)baclofen was mediated by GABA/sub B/ receptor activation, since it was blocked by GABA/sub B/ antagonist phaclofen, but notmore » by bicuculline. Further, pertussis toxin also blocked the ability of baclofen or depolarizing action to affect Ca/sup 2 +/-activated K/sup +/-channels. These results suggest that baclofen inhibits Ca/sup 2 +/-activated K/sup +/-channels in synaptosomes and these channels are regulated by G-proteins. This assay may provide an ideal in vitro model to study GABA/sub B/ receptor pharmacology.« less

  19. Simultaneous presence of (Si{sub 3}O{sub 10}){sup 8−} and (Si{sub 2}O{sub 7}){sup 6−} groups in new synthetic mixed sorosilicates: BaY{sub 4}(Si{sub 2}O{sub 7})(Si{sub 3}O{sub 10}) and isotypic compounds, studied by single-crystal X-ray diffraction, Raman spectroscopy and DFT calculations

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

    Wierzbicka-Wieczorek, Maria, E-mail: maria.wierzbicka-wieczorek@uni-jena.de; Többens, Daniel M.; Kolitsch, Uwe

    2013-11-15

    Three new, isotypic silicate compounds, BaY{sub 4}(Si{sub 2}O{sub 7})(Si{sub 3}O{sub 10}), SrYb{sub 4}(Si{sub 2}O{sub 7})(Si{sub 3}O{sub 10}) and SrSc{sub 4}(Si{sub 2}O{sub 7})(Si{sub 3}O{sub 10}), were synthesized using high-temperature flux growth techniques, and their crystal structures were solved from single-crystal X-ray intensity data: monoclinic, P2{sub 1}/m, with a=5.532(1)/5.469(1)/5.278(1), b=19.734(4)/19.447(4)/19.221(4), c=6.868(1)/6.785(1)/6.562(1) Å, β=106.53(3)/106.20(3)/106.50(3)°, V=718.8(2)/693.0(2)/638.3(2) Å{sup 3}, R(F)=0.0225/0.0204/0.0270, respectively. The topology of the novel structure type contains isolated horseshoe-shaped Si{sub 3}O{sub 10} groups (Si–Si–Si=93.15–95.98°), Si{sub 2}O{sub 7} groups (Si–O{sub bridge}–Si=180°, symmetry-restricted) and edge-sharing M(1)O{sub 6} and M(2)O{sub 6} octahedra. Single-crystal Raman spectra of the title compounds were measured and compared with Raman spectroscopicmore » data of chemically and topologically related disilicates and trisilicates, including BaY{sub 2}(Si{sub 3}O{sub 10}) and SrY{sub 2}(Si{sub 3}O{sub 10}). The band assignments are supported by additional theoretical calculation of Raman vibrations by DFT methods. - Graphical abstract: View of BaY{sub 4}(Si{sub 2}O{sub 7})(Si{sub 3}O{sub 10}) along [100], showing zigzag chains and the tri- and disilicate groups. The unit cell is outlined. Display Omitted - Highlights: • We report a novel interesting crystal structure type for mixed sorosilicates containing Y, Yb, and Sc. • Synthesis of such mixed sorosilicates is possible by a high-temperature flux-growth technique. • Calculation of Raman vibrations by advanced DFT methods allows a considerably improved interpretation of measured Raman spectra.« less

  20. Antiedematogenic and antioxidant properties of high molecular weight protein sub-fraction of Calotropis procera latex in rat.

    PubMed

    Chaudhary, Priyanka; de Araújo Viana, Carolina; Ramos, Marcio V; Kumar, Vijay L

    2015-03-01

    The aim was to evaluate the effect of high molecular weight protein fraction of Calotropis procera latex on edema formation and oxidative stress in carrageenan-induced paw inflammation. A sub-plantar injection of carrageenan was given to induce edema in the hind paw of the rat. The inhibitory effect of high molecular weight protein fraction of C. procera latex was evaluated following intravenous administration (5 and 25 mg/kg body weight) and was compared with that of diclofenac given orally (5 mg/kg). The levels of reduced glutathione (GSH), thiobarbituric acid reactive substances (TBARS) and myeloperoxidase (MPO) were measured in the inflamed paw tissue at the end of the study. The high molecular weight protein fraction obtained from the latex of C. procera produced a dose-dependent inhibition of edema formation that was accompanied by normalization of levels of oxidative stress markers (GSH and TBARS) and MPO, a marker for neutrophils in the paw tissue. The high molecular weight protein fraction of C. procera latex ameliorates acute inflammation in the paw through its antioxidant effect.

  1. Protein Turnover and Metabolism in the Elderly Intensive Care Unit Patient.

    PubMed

    Phillips, Stuart M; Dickerson, Roland N; Moore, Frederick A; Paddon-Jones, Douglas; Weijs, Peter J M

    2017-04-01

    Many intensive care unit (ICU) patients do not achieve target protein intakes particularly in the early days following admittance. This period of iatrogenic protein undernutrition contributes to a rapid loss of lean, in particular muscle, mass in the ICU. The loss of muscle in older (aged >60 years) patients in the ICU may be particularly rapid due to a perfect storm of increased catabolic factors, including systemic inflammation, disuse, protein malnutrition, and reduced anabolic stimuli. This loss of muscle mass has marked consequences. It is likely that the older patient is already experiencing muscle loss due to sarcopenia; however, the period of stay in the ICU represents a greatly accelerated period of muscle loss. Thus, on discharge, the older ICU patient is now on a steeper downward trajectory of muscle loss, more likely to have ICU-acquired muscle weakness, and at risk of becoming sarcopenic and/or frail. One practice that has been shown to have benefit during ICU stays is early ambulation and physical therapy (PT), and it is likely that both are potent stimuli to induce a sensitivity of protein anabolism. Thus, recommendations for the older ICU patient would be provision of at least 1.2-1.5 g protein/kg usual body weight/d, regular and early utilization of ambulation (if possible) and/or PT, and follow-up rehabilitation for the older discharged ICU patient that includes rehabilitation, physical activity, and higher habitual dietary protein to change the trajectory of ICU-mediated muscle mass loss and weakness.

  2. C-Myc Protein-Protein and Protein-DNA Interactions: Targets for Therapeutic Intervention.

    DTIC Science & Technology

    1997-09-01

    including those of the Myc family. In fact, members of different bHLH protein subgroups, including the Myc proteins, are characterized by conserved BR...important functional consequences, and they provide insights into how different bHLH proteins can act on different targets. The zinc finger protein...roles for a number of BR residues which do not contact bases, yet are conserved within different bHLH protein sub- families (Benezra et al. 1990), and

  3. Electronic structure of Pr{sub 1{minus}x}Y{sub x}Ba{sub 2}Cu{sub 3}O{sub y} (x=0, 0.5, and 1.0)

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

    Kakinuma, K.; Fueki, K.

    1997-08-01

    In order to elucidate the reason why PrBa{sub 2}Cu{sub 3}O{sub y}is not a superconductor, we examined the Pr valence and measured the oxygen nonstoichiometry and the conductivity at temperatures up to 1200 K for three kinds of oxides, PrBa{sub 2}Cu{sub 3}O{sub y}, (Pr{sub 0.5}Y{sub 0.5})Ba{sub 2}Cu{sub 3}O{sub y}, and YBa{sub 2}Cu{sub 3}O{sub y}. The valence of Pr was found to be +3. Any difference was not found in oxygen nonstoichiometry and conductivity among three kinds of oxides. We analyzed the data of oxygen nonstoichiometry on the basis of defect thermodynamics and calculated the numbers of Cu{sup +}, Cu{sup 2+}, andmore » Cu{sup 3+} ions in the unit cell as a function of y. The number of Cu{sup 3+} ions (the amount of holes) was found to be proportional to ({Delta}y){sup 1.6}({Delta}y=y{minus}6.0), whereas the conductivity was found to be proportional to ({Delta}y){sup 3.2} in these oxides. We interpreted the remarkable increase of {sigma} with {Delta}y as an evidence of the increase of both mobility and hole concentration with {Delta}y. At high temperatures, we detected the conductivity minimum {sigma}{sub min} which was found in the log{sub 10}{sigma}{minus}log{sub 10}P{sub O{sub 2}} plot at constant temperatures. From the slope of the Arrhenius plot for {sigma}{sub min}, the band gap was determined to be 1.21, 1.32, and 1.37 eV for PrBa{sub 2}Cu{sub 3}O{sub y}, (Pr{sub 0.5}Y{sub 0.5})Ba{sub 2}Cu{sub 3}O{sub y} and YBa{sub 2}Cu{sub 3}O{sub y}, respectively. We determined the conductivity of the same oxygen content as a function of temperature from 4.2 to 1200 K. The energy gap {Delta}E between the acceptor level and the top of the valence band was calculated from the slope of the Arrhenius plot for conductivity. {Delta}E for superconducting YBa{sub 2}Cu{sub 3}O{sub y} and (Pr{sub 0.5}Y{sub 0.5})Ba{sub 2}Cu{sub 3}O{sub y} were zero at 300 K but that for nonsuperconducting PrBa{sub 2}Cu{sub 3}O{sub y} was 20 meV at 100 K even for y=6.93. (Abstract

  4. HT.sub.m4 methods of treatment and assays, agonists and antagonists

    DOEpatents

    Lim, Bing; Adra, Chaker N.; Lelias, Jean-Michel

    1999-01-01

    The invention relates to a recombinant DNA molecule which encodes a HT.sub.m4 protein, a transformed host cell which has been stably transfected with a DNA molecule which encodes a HT.sub.m4 protein and a recombinant HT.sub.m4 protein. The invention also relates to a method for detecting the presence of a hereditary atopy.

  5. Surfactant protein D attenuates sub-epithelial fibrosis in allergic airways disease through TGF-β.

    PubMed

    Ogawa, Hirohisa; Ledford, Julie G; Mukherjee, Sambuddho; Aono, Yoshinori; Nishioka, Yasuhiko; Lee, James J; Izumi, Keisuke; Hollingsworth, John W

    2014-11-29

    Surfactant protein D (SP-D) can regulate both innate and adaptive immunity. Recently, SP-D has been shown to contribute to the pathogenesis of airway allergic inflammation and bleomycin-induced pulmonary fibrosis. However, in allergic airways disease, the role of SP-D in airway remodeling remains unknown. The objective of this study was to determine the contribution of functional SP-D in regulating sub-epithelial fibrosis in a mouse chronic house dust mite model of allergic airways disease. C57BL/6 wild-type (WT) and SP-D-/- mice (C57BL/6 background) were chronically challenged with house dust mite antigen (Dermatophagoides pteronyssinus, Dp). Studies with SP-D rescue and neutralization of TGF-β were conducted. Lung histopathology and the concentrations of collagen, growth factors, and cytokines present in the airspace and lung tissue were determined. Cultured eosinophils were stimulated by Dp in presence or absence of SP-D. Dp-challenged SP-D-/- mice demonstrate increased sub-epithelial fibrosis, collagen production, eosinophil infiltration, TGF-β1, and IL-13 production, when compared to Dp-challenged WT mice. By immunohistology, we detected an increase in TGF-β1 and IL-13 positive eosinophils in SP-D-/- mice. Purified eosinophils stimulated with Dp produced TGF-β1 and IL-13, which was prevented by co-incubation with SP-D. Additionally, treatment of Dp challenged SP-D-/- mice with exogenous SP-D was able to rescue the phenotypes observed in SP-D-/- mice and neutralization of TGF-β1 reduced sub-epithelial fibrosis in Dp-challenged SP-D-/- mice. These data support a protective role for SP-D in the pathogenesis of sub-epithelial fibrosis in a mouse model of allergic inflammation through regulation of eosinophil-derived TGF-β.

  6. A first-principles study on the B{sub 5}O{sub 5}{sup +/0} and B{sub 5}O{sub 5}{sup −} clusters: The boron oxide analogs of C{sub 6}H{sub 5}{sup +/0} and CH{sub 3}Cl

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

    Tian, Wen-Juan; You, Xue-Rui; Ou, Ting

    2015-08-14

    The concept of boronyl (BO) and the BO/H isolobal analogy build an interesting structural link between boron oxide clusters and hydrocarbons. Based upon global-minimum searches and first-principles electronic structural calculations, we present here the perfectly planar C{sub 2v} B{sub 5}O{sub 5}{sup +} (1, {sup 1}A{sub 1}), C{sub 2v} B{sub 5}O{sub 5} (2, {sup 2}A{sub 1}), and tetrahedral C{sub s} B{sub 5}O{sub 5}{sup −} (3, {sup 1}A′) clusters, which are the global minima of the systems. Structural and molecular orbital analyses indicate that C{sub 2v} B{sub 5}O{sub 5}{sup +} (1) [B{sub 3}O{sub 3}(BO){sub 2}{sup +}] and C{sub 2v} B{sub 5}O{sub 5}more » (2) [B{sub 3}O{sub 3}(BO){sub 2}] feature an aromatic six-membered boroxol (B{sub 3}O{sub 3}) ring as the core with two equivalent boronyl terminals, similar to the recently reported boronyl boroxine D{sub 3h} B{sub 6}O{sub 6} [B{sub 3}O{sub 3}(BO){sub 3}]; whereas C{sub s} B{sub 5}O{sub 5}{sup −} (3) [B(BO){sub 3}(OBO){sup −}] is characterized with a tetrahedral B{sup −} center, terminated with three BO groups and one OBO unit, similar to the previously predicted boronyl methane T{sub d} B{sub 5}O{sub 4}{sup −} [B(BO){sub 4}{sup −}]. Alternatively, the 1–3 clusters can be viewed as the boron oxide analogs of phenyl cation C{sub 6}H{sub 5}{sup +}, phenyl radical C{sub 6}H{sub 5}, and chloromethane CH{sub 3}Cl, respectively. Chemical bonding analyses also reveal a dual three-center four-electron (3c-4e) π hyperbond in C{sub s} B{sub 5}O{sub 5}{sup −} (3). The infrared absorption spectra of B{sub 5}O{sub 5}{sup +} (1), B{sub 5}O{sub 5} (2), and B{sub 5}O{sub 5}{sup −} (3) and anion photoelectron spectrum of B{sub 5}O{sub 5}{sup −} (3) are predicted to facilitate their forthcoming experimental characterizations. The present work completes the B{sub n}O{sub n}{sup +/0/−} series for n = 1–6 and enriches the analogous relationship between boron oxides and hydrocarbons.« less

  7. The crystal structures of Ni{sub 3+x}Sn{sub 4}Zn and Ni{sub 6+x}Sn{sub 8}Zn and their structural relations to Ni{sub 3+x}Sn{sub 4}, NiSn and Ni{sub 5−δ}ZnSn{sub 4}

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

    Schmetterer, Clemens, E-mail: clemens.schmetterer@univie.ac.at; Effenberger, Herta Silvia; Rajamohan, Divakar

    2016-06-15

    The crystal structures of two new compounds were determined from single-crystal X-ray diffraction measurements: Ni{sub 3+x}Sn{sub 4}Zn, (x~1.35, a=7.110(2) Å, b=4.123(1) Å, c=10.346(3) Å, β=90.23(2)°, space group I2/m, Z=2. R1=0.025, wR2=0.059 for 748 unique reflections, 35 variable parameters) and Ni{sub 6+x}Sn{sub 8}Zn, x~1.35 (a=12.379(3) Å, b=4.095(1) Å, c=12.155(3) Å, β=116.25(3)°, space group C2/m, Z=2. R1=0.026, wR2=0.052 for 1346 unique reflections, 60 variable parameters). In addition, a structural refinement was performed for Ni{sub 3+x}Sn{sub 4}, x~0.13 (a=12.264(3) Å, b=4.066(1) Å, c=5.223(2) Å, β=104.85(3)°, space group C2/m, Z=2. R1=0.019, wR2=0.046 for 617 unique reflections, 29 variable parameters). The three compounds show pronouncedmore » similarities among each other as well as to the crystal structures of surrounding binary Ni–Sn and ternary Ni–Sn–Zn compounds. In particular, the two new compounds form a homologous series with Ni{sub 3+x}Sn{sub 4}, x~0.13. They contain “Ni{sub 4}Sn{sub 4}” and “Ni{sub 2}Sn{sub 4}” building blocks which by different interconnection build up the distinct structures. Topological relations with NiSn and Ni{sub 5−δ}Sn{sub 4}Zn, δ~0.25 are evident. - Graphical abstract: Projection of the structure of Ni{sub 6+x}ZnSn{sub 8}, x~1.35 and constituent building blocks. Display Omitted - Highlights: • The crystal structures of Ni{sub 6+x}Sn{sub 8}Zn and Ni{sub 3+x}Sn{sub 4}Zn were determined using single crystal XRD. • Topological relations to Ni–Sn and Ni–Sn–Zn compounds were established and discussed. • Common structural units were identified and their interconnection patterns described.« less

  8. Resistance of Adenoviral DNA Replication to Aphidicolin Is Dependent on the 72-Kilodalton DNA-Binding Protein

    PubMed Central

    Foster, David A.; Hantzopoulos, Petros; Zubay, Geoffrey

    1982-01-01

    Aphidicolin is a highly specific inhibitor of DNA polymerase α and has been most useful for assessing the role of this enzyme in various replication processes (J. A. Huberman, Cell 23:647-648, 1981). Both nuclear DNA replication and simian virus 40 DNA replication are highly sensitive to this drug (Krokan et al., Biochemistry 18:4431-4443, 1979), whereas mitochondrial DNA synthesis is completely insensitive (Zimmerman et al., J. Biol. Chem. 255:11847-11852, 1980). Adenovirus DNA replication is sensitive to aphidicolin, but only at much higher concentrations. These patterns of sensitivity are seen both in vivo and in vitro (Krokan et al., Biochemistry 18:4431-4443, 1979). A temperature-sensitive mutant of adenovirus type 5 known as H5ts125 is able to complete but not initiate new rounds of replication at nonpermissive temperatures (P. C. van der Vliet and J. S. Sussenbach, Virology 67:415-426, 1975). When cells infected with H5ts125 were shifted from permissive (33°C) to nonpermissive (41°C) conditions, the residual DNA synthesis (elongation) showed a striking increase in sensitivity to aphidicolin. The temperature-sensitive mutation of H5ts125 is in the gene for the 72-kilodalton single-stranded DNA-binding protein. This demonstrated that the increased resistance to aphidicolin shown by adenovirus DNA replication was dependent on that protein. It also supports an elongation role for both DNA polymerase α and the 72-kilodalton single-stranded DNA-binding protein in adenovirus DNA replication. Further support for an elongation role of DNA polymerase α came from experiments with permissive temperature conditions and inhibiting levels of aphidicolin in which it was shown that newly initiated strands failed to elongate to completion. Images PMID:6809958

  9. Metal–organic frameworks assembled from lanthanide and 2,5-pyridinedicaboxylate with cubane-like [Ln{sub 4}(OH){sub 4}] building units

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

    Abdelbaky, Mohammed S.M.; Amghouz, Zakariae, E-mail: amghouz.uo@uniovi.es; Servicios Científico-Técnicos, University of Oviedo—CINN, Oviedo 33006

    Lanthanide–organic frameworks based on 2,5-pyridinedicaboxylate (25p) ligand, formulated as [Yb{sub 4}(OH){sub 4}(25p){sub 4}(H{sub 2}O){sub 3}]·H{sub 2}O (25pYb), [Y{sub 4}(OH){sub 4}(25p){sub 4}(H{sub 2}O){sub 3}]·H{sub 2}O (25pY-1) and [Y{sub 6}(OH){sub 8}(25p){sub 5}(H{sub 2}O){sub 2}] (25pY-2), have been obtained as single phases under hydrothermal conditions. 25pYb and 25pY-1 are isostructural, and crystallize in the triclinic space group, P-1, with a=8.6075(5) Å, b=14.8478(7) Å, c=15.9164(9) Å, α=86.277(4)°, β=80.196(5)°, γ=81.785(4)°, and a=8.7166(6) Å, b=14.966(1) Å, c=15.966(1) Å, α=86.260(6)°, β=80.036(6)°, γ=81.599(6)°, respectively. 25pY-2 crystallizes in the monoclinic space group, P2{sub 1}/c, with a=24.9117(17) Å, b=13.7340(8) Å, c=14.3385(10) Å, β=100.551(7)°. 25pYb and 25pY-2 have been structurally characterizedmore » by single-crystal X-ray diffraction. The 25pYb structure is based on tetranuclear cubane-like [Yb{sub 4}(OH){sub 4}]{sup 8+} clusters, which are interconnected to eight neighbouring clusters through teen surrounding 25p ligands leading to neutral 3D framework, while the structure of 25pY-2 is based on two independent cuban-like [Y{sub 4}(OH){sub 4}]{sup 8+} clusters, which are joined together through Y1 cation leading to the formation of hexanuclear [Y{sub 6}(OH){sub 8}]{sup 10+} clusters, which in turn are joined via Y2 cation resulting in infinite inorganic chain extending along c-axis, and each chain is interconnected to six adjacent chains through 25p ligands leading finally to 3D framework. The luminescence properties of Eu{sup 3+} and Tb{sup 3+} doped 25pY-1 and 25pY-2 compounds have also been investigated. All materials has been characterized by powder X-ray diffraction, thermal analyses (TG–SDTA–MS), FTIR spectroscopy, C–H–N elemental analysis, scanning electron microscopy (SEM-EDX), and powder X-ray thermodiffraction. - Graphical abstract: Nowadays, lanthanide–organic frameworks (LOFs

  10. Preliminary crystallographic analysis of the major capsid protein P2 of the lipid-containing bacteriophage PM2

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

    Abrescia, Nicola G. A.; Kivelä, Hanna M.; Grimes, Jonathan M.

    2005-08-01

    The viral capsid protein P2 of bacteriophage PM2 has been crystallized. Preliminary X-ray analysis demonstrates the position and orientation of the two trimers in the asymmetric unit. PM2 (Corticoviridae) is a dsDNA bacteriophage which contains a lipid membrane beneath its icosahedral capsid. In this respect it resembles bacteriophage PRD1 (Tectiviridae), although it is not known whether the similarity extends to the detailed molecular architecture of the virus, for instance the fold of the major coat protein P2. Structural analysis of PM2 has been initiated and virus-derived P2 has been crystallized by sitting-nanodrop vapour diffusion. Crystals of P2 have been obtainedmore » in space group P2{sub 1}2{sub 1}2, with two trimers in the asymmetric unit and unit-cell parameters a = 171.1, b = 78.7, c = 130.1 Å. The crystals diffract to 4 Å resolution at the ESRF BM14 beamline (Grenoble, France) and the orientation of the non-crystallographic threefold axes, the spatial relationship between the two trimers and the packing of the trimers within the unit cell have been determined. The trimers form tightly packed layers consistent with the crystal morphology, possibly recapitulating aspects of the arrangement of subunits in the virus.« less

  11. The crystal structure of the interrupted framework silicate K{sub 9.6}Ca{sub 1.2}Si{sub 12}O{sub 30} determined from laboratory X-ray diffraction data

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

    Kahlenberg, V.; Kaindl, R.; Christian-Doppler-Laboratory for Advanced Hard Coatings at the Institute of Mineralogy and Petrography, University of Innsbruck, Innrain 52, A-6020 Innsbruck

    2006-07-15

    The crystal structure of a potassium calcium silicate with composition K{sub 9.6}Ca{sub 1.2}Si{sub 12}O{sub 30} (or K{sub 8}CaSi{sub 10}O{sub 25}) has been solved by direct methods aided by distance least squares optimization from laboratory X-ray powder diffraction data. The trigonal compound adopts the non-centrosymmetric space group R3c with the following basic crystallographic data: a=11.13623(5)A, c=21.9890(2)A, V=2361.63(2)A{sup 3}, Z=3, D{sub calc}=2.617gcm{sup -3}. The crystal structure can be classified as an interrupted framework with exclusively Q{sup 3}-units. It can be thought of as being built from layers parallel to (001) containing isolated six-membered tetrahedral rings in UDUDUD conformation. Corner sharing of tetrahedramore » belonging to adjacent sheets results in a tetrahedral framework. The framework density of the structure is 15.2 T-atoms/1000A{sup 3}. The coordination sequences are identical for both silicon atoms in the asymmetric unit: 3-6-11-20-32-46-60-80-102-122. The vertex symbols for the two tetrahedral centers are 10{sub 2}.10{sub 2}.6{sub 1}. Topologically, the structure can be described as an Archimedean three-dimensional 3-connected net. It can be derived from the diamond or cristobalite net by removing 20% of the knots. Charge compensation in the structure is achieved by the incorporation of mono- and divalent M-cations (M: K, Ca). These extra-framework ions are coordinated by six to nine oxygen ligands. Ca/K distributions for the five symmetrically independent M-sites were obtained from a combination of bond distance considerations, site occupancy refinements and the bulk chemical composition. The structural characterization is completed by a detailed Raman spectroscopic study. Furthermore, possible implications of the structural chemistry of interrupted framework silicates for the field of silicate glass research are addressed.« less

  12. Adenovirus-mediated suppression of HMGI(Y) protein synthesis as potential therapy of human malignant neoplasias

    PubMed Central

    Scala, Stefania; Portella, Giuseppe; Fedele, Monica; Chiappetta, Gennaro; Fusco, Alfredo

    2000-01-01

    High mobility group I (HMGI) proteins are overexpressed in several human malignant tumors. We previously demonstrated that inhibition of HMGI synthesis prevents thyroid cell transformation. Here, we report that an adenovirus carrying the HMGI(Y) gene in an antisense orientation (Ad-Yas) induced programmed cell death of two human thyroid anaplastic carcinoma cell lines (ARO and FB-1), but not normal thyroid cells. The Ad-Yas virus led to death of lung, colon, and breast carcinoma cells. A control adenovirus carrying the lacZ gene did not inhibit the growth of either normal or neoplastic cells. Ad-Yas treatment of tumors induced in athymic mice by ARO cells caused a drastic reduction in tumor size. Therefore, suppression of HMGI(Y) protein synthesis by an HMGI(Y) antisense adenoviral vector may be a useful treatment strategy in a variety of human malignant neoplasias, in which HMGI(Y) gene overexpression is a general event. PMID:10759549

  13. Synthesis, characterization and optical properties of NH{sub 4}Dy(PO{sub 3}){sub 4}

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

    Chemingui, S.; Ferhi, M., E-mail: ferhi.mounir@gmail.com; Horchani-Naifer, K.

    2014-09-15

    Polycrystalline powders of NH{sub 4}Dy(PO{sub 3}){sub 4} polyphosphate have been grown by the flux method. This compound was found to be isotopic with NH{sub 4}Ce(PO{sub 3}){sub 4} and RbHo(PO{sub 3}){sub 4}. It crystallizes in the monoclinic space group P2{sub 1/n} with unit cell parameters a=10.474(6) Å, b=9.011(4) Å, c=10.947(7) Å and β=106.64(3)°. The title compound has been transformed to triphosphate Dy(PO{sub 3}){sub 3} after calcination at 800 °C. Powder X-ray diffraction, infrared and Raman spectroscopies and the differential thermal analysis have been used to identify these materials. The spectroscopic properties have been investigated through absorption, excitation, emission spectra and decaymore » curves of Dy{sup 3+} ion in both compounds at room temperature. The emission spectra show the characteristic emission bands of Dy{sup 3+} in the two compounds, before and after calcination. The integrated emission intensity ratios of the yellow to blue (I{sub Y}/I{sub B}) transitions and the chromaticity properties have been determined from emission spectra. The decay curves are found to be double-exponential. The non-exponential behavior of the decay rates was related to the resonant energy transfer as well as cross-relaxation between the donor and acceptor Dy{sup 3+} ions. The determined properties have been discussed as function of crystal structure of both compounds. They reveal that NH{sub 4}Dy(PO{sub 3}){sub 4} is promising for white light generation but Dy(PO{sub 3}){sub 3} is potential candidates in field emission display (FED) and plasma display panel (PDP) devices. - Graphical abstract: The CIE color coordinate diagrams showing the chromatic coordinates of Dy{sup 3+} luminescence in NH{sub 4}Dy(PO{sub 3}){sub 4} and Dy(PO{sub 3}){sub 3}. - Highlights: • The polycrystalline powders of NH{sub 4}Dy(PO{sub 3}){sub 4} and Dy(PO{sub 3}){sub 3} are synthesized. • The obtained powders are characterized. • The spectroscopic properties of Dy

  14. Quadruplexes of human telomere dG{sub 3}(TTAG{sub 3}){sub 3} sequences containing guanine abasic sites

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

    Skolakova, Petra; Bednarova, Klara; Vorlickova, Michaela

    Research highlights: {yields} Loss of a guanine base does not hinder the formation of G-quadruplex of human telomere sequence. {yields} Each depurination strongly destabilizes the quadruplex of dG{sub 3}(TTAG{sub 3}){sub 3} in NaCl and KCl. {yields} Conformational change of the abasic analogs of dG{sub 3}(TTAG{sub 3}){sub 3} is inhibited in KCl. {yields} The effects abasic sites may affect telomere-end structures in vivo. -- Abstract: This study was performed to evaluate how the loss of a guanine base affects the structure and stability of the three-tetrad G-quadruplex of 5'-dG{sub 3}(TTAG{sub 3}){sub 3}, the basic quadruplex-forming unit of the human telomere DNA.more » None of the 12 possible abasic sites hindered the formation of quadruplexes, but all reduced the thermodynamic stability of the parent quadruplex in both NaCl and KCl. The base loss did not change the Na{sup +}-stabilized intramolecular antiparallel architecture, based on CD spectra, but held up the conformational change induced in dG{sub 3}(TTAG{sub 3}){sub 3} in physiological concentration of KCl. The reduced stability and the inhibited conformational transitions observed here in vitro for the first time may predict that unrepaired abasic sites in G-quadruplexes could lead to changes in the chromosome's terminal protection in vivo.« less

  15. Overexpression, purification and crystallization of a choline-binding protein CbpI from Streptococcus pneumoniae

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

    Paterson, Neil G., E-mail: neison@chem.gla.ac.uk; Riboldi-Tunicliffe, Alan; Mitchell, Timothy J.

    2006-07-01

    The choline-binding protein CbpI from S. pneumoniae has been purified and crystallized and diffraction data have been collected to 3.5 Å resolution. The choline-binding protein CbpI from Streptococcus pneumoniae is a 23.4 kDa protein with no known function. The protein has been successfully purified initially using Ni–NTA chromatography and to homogeneity using Q-Sepharose ion-exchange resin as an affinity column. CbpI was crystallized using PEG 3350 as a precipitant and X-ray crystallographic analysis showed that the crystals belonged to the tetragonal space group P4, with unit-cell parameters a = b = 83.31, c = 80.29 Å, α = β = γmore » = 90°. The crystal contains two molecules in the asymmetric unit with a solvent content of 55.7% (V{sub M} = 2.77 Å{sup 3} Da{sup −1}) and shows a diffraction limit of 3.5 Å.« less

  16. pH shift assembly of adenoviral serotype 5 capsid protein nanosystems for enhanced delivery of nanoparticles, proteins and nucleic acids.

    PubMed

    Rao, Vidhya R; Upadhyay, Arun K; Kompella, Uday B

    2013-11-28

    Empty adenovirus serotype 5 (Ad5) capsids devoid of viral genome were developed as a novel delivery system for nanoparticles, proteins, and nucleic acids. Ad5 capsids of 110 nm diameter undergo an increase in particle size to 1637 nm in 1mM acetic acid at pH4.0 and then shrink to 60 nm, following pH reversal to 7.4. These pH shifts induced reversible changes in capsid zeta potential and secondary structure and irreversible changes in tertiary structure of capsid proteins. Using pH shift dependent changes in capsid size and structure, 20 nm fluorescent nanoparticles, FITC-BSA, and Alexa Fluor® 488 conjugated siRNA were encapsulated with high efficiency in Ad5 capsids, as confirmed by electron microscopy and/or flow cytometry. HEK cell uptake with capsid delivery system was 7.8-, 7.4-, and 2.9-fold greater for nanoparticles, FITC-BSA, and Alexa-siRNA, respectively, when compared to plain solutes. Physical mixtures of capsids and fluorescent solutes exhibited less capsid associated fluorescence intensity and cell uptake. Further, unlike physical mixture, pH shift assembled Ad5 capsids protected siRNA from RNase degradation. Ad5 capsids before and after pH shift exhibited endolysosomal escape. Thus, empty Ad5 capsids can encapsulate a variety of solutes based on pH shift assembly, resulting in enhanced cellular delivery. © 2013. Published by Elsevier B.V. All rights reserved.

  17. Demonstration of natural gas reburn for NO{sub x} emissions reduction at Ohio Edison Company`s cyclone-fired Niles Plant Unit Number 1

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

    Borio, R.W.; Lewis, R.D.; Koucky, R.W.

    1996-04-01

    Electric utility power plants account for about one-third of the NO{sub x} and two-thirds of the SO{sub 2} emissions in the US cyclone-fired boilers, while representing about 9% of the US coal-fired generating capacity, emit about 14% of the NO{sub x} produced by coal-fired utility boilers. Given this background, the Environmental Protection Agency, the Gas Research Institute, the Electric Power Research Institute, the Pittsburgh Energy Technology Center, and the Ohio Coal Development Office sponsored a program led by ABB Combustion Engineering, Inc. (ABB-CE) to demonstrate reburning on a cyclone-fired boiler. Ohio Edison provided Unit No. 1 at their Niles Stationmore » for the reburn demonstration along with financial assistance. The Niles Unit No. 1 reburn system was started up in September 1990. This reburn program was the first full-scale reburn system demonstration in the US. This report describes work performed during the program. The work included a review of reburn technology, aerodynamic flow model testing of reburn system design concepts, design and construction of the reburn system, parametric performance testing, long-term load dispatch testing, and boiler tube wall thickness monitoring. The report also contains a description of the Niles No. 1 host unit, a discussion of conclusions and recommendations derived from the program, tabulation of data from parametric and long-term tests, and appendices which contain additional tabulated test results.« less

  18. Absolute configurations of organometallic compounds. III. Structure and absolute configuration of the square-pyramidal complex ((+)/sub 579/-(C/sub 5/H/sub 5/)Mo(CO)/sub 2/(NN*))PF/sub 6/(NN* = Schiff base derived from pyridine-2-carbaldehyde and (S)-(-)-. cap alpha. -phenylethylamine)

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

    Bernal, I.; LaPlaca, S.J.; Korp, J.

    The structure of (+)/sub 579/-(eta/sup 5/-C/sub 5/H/sub 5/Mo(CO)/sub 2/(NN*))PF/sub 6/ with NN* = the Schiff base derived from pyridine-2-carbaldehyde and (S)-(--)-..cap alpha..-phenylethylamine was determined using standard single-crystal x-ray diffraction methods. The absolute configuration was determined by refinement of the data using the anomalous scattering contributions of Mo and P to a final R(F) = 0.056 for 2634 independent reflections having I greater than 3 sigma (I). The substance crystallizes in the space group P2/sub 1/2/sub 1/2/sub 1/ with unit cell dimensions of a = 12.249 (4), b = 9.236 (3), and c = 20.692 (9) A and Z = 4more » molecules/unit cell. The square-pyramidal coordination of the Mo atom is defined by two carbonyl carbons and two Schiff base nitrogens occupying the four basal plane sites and the five carbons of the eta/sup 5/-C/sub 5/H/sub 5/ ligand in the axial position. The Mo--ligand distances and the bond lengths and angles within the ligands are normal and compare closely with those of recent structure determinations of comparable precision. The Mo atom is 0.95 A above the plane formed by the four basal plane ligands. The conformation of the (S)-..cap alpha..-phenylethyl group with respect to the ligand plane, defined by the pyridine ring, the imine system, and the Mo atom, is discussed. The configuration at the metal atom in the (+)/sub 579/ isomer is specified as (S). The PF/sub 6//sup -/ anion executes large amplitude torsional motion in the lattice, as is commonly the case for this anion when not hydrogen bonded.« less

  19. Study on the site preference of Ca in superconducting oxides Bi{sub 2}Sr{sub 2−x}Ca{sub x}CuO{sub 6+δ} (0.1 ≤ x ≤ 1.0)

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

    Sun, B.Z.; Zhou, S.L.; Wang, H.

    2014-01-15

    A series of compound with the nominal composition of Bi{sub 2}Sr{sub 2−x}Ca{sub x}CuO{sub 6+δ} (x = 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0) were synthesized by the sol–gel method. Constituent phases and crystal structure of samples were analyzed by X-ray diffraction. It can be found that the Ca-doped Bi-2201 system was composed of Bi-2201 phase containing Ca and a small quantity of Bi{sub 16}(Sr,Ca){sub 14}O{sub 38}. For Bi-2201 unit cell containing Ca, chemical component and site preference of Ca atoms were characterized systematically by transmission electron microscopy. With the introduction of Ca atoms, Sr-sites have been occupiedmore » partially by Ca{sup 2+} in Bi-2201 unit cell, which leads to a decrease in the lattice parameters c and b of the Bi-2201 phase when the Ca-content x is below 0.6. Two types of new orthorhombic lattices are formed in the substitution. One is a lattice with space group Pma2 as the two nearest neighbor Sr-sites in the same Sr–O layer are occupied by Ca{sup 2+}. Its lattice parameters can be characterized as a = 5.402 Å, b = 5.313 Å and c = 24.272 Å, respectively. When two nearest Sr ions of the second neighboring Sr–O layers are replaced by Ca{sup 2+} ions, the lattice with the space group Pmn2{sub 1} can be formed. Its lattice parameters are close to that of the previous. The modulation vector is lying in the a*–c* plane in the two new orthorhombic lattices (Pma2 and Pmn2{sub 1}). Bi/Ca-2201 lattice (with Ca) and Bi-2201 lattice (without Ca) coexist in the same Bi{sub 2}Sr{sub 2−x}Ca{sub x}CuO{sub 6}+{sub δ} grain, which can be described as an intergrowth structure.« less

  20. Lacritin and Other New Proteins of the Lacrimal Functional Unit

    PubMed Central

    McKown, Robert L.; Wang, Ningning; Raab, Ronald W.; Karnati, Roy; Zhang, Yinghui; Williams, Patricia B.; Laurie, Gordon W.

    2009-01-01

    The lacrimal functional unit (LFU) is defined by the 2007 International Dry Eye WorkShop as ‘an integrated system comprising the lacrimal glands, ocular surface (cornea, conjunctiva and meibomian glands) and lids, and the sensory and motor nerves that connect them’. The LFU maintains a healthy ocular surface primarily through a properly functioning tear film that provides protection, lubrication, and an environment for corneal epithelial cell renewal. LFU cells express thousands of proteins. Over two hundred new LFU proteins have been discovered in the last decade. Lacritin is a new LFU-specific growth factor in human tears that flows through ducts to target corneal epithelial cells on the ocular surface. When applied topically in rabbits, lacritin appears to increase the volume of basal tear secretion. Lacritin is one of only a handful of tear proteins preliminarily reported to be downregulated in blepharitis and in two dry eye syndromes. Computational analysis predicts an ordered C-terminal domain that binds the corneal epithelial cell surface proteoglycan syndecan-1 (SDC1) and is required for lacritin’s low nanomolar mitogenic activity. The lacritin binding site on the N-terminus of SDC1 is exposed by heparanase. Heparanase is constitutively expressed by the corneal epithelium and appears to be a normal constituent of tears. Binding triggers rapid signaling to downstream NFAT and mTOR. A wealth of other new proteins, originally designated as hypothetical when first identified by genomic sequencing, are expressed by the human LFU including: ALS2CL, ARHGEF19, KIAA1109, PLXNA1, POLG, WIPI1 and ZMIZ2. Their demonstrated or implied roles in human genetic disease or basic cellular functions are fuel for new investigation. Addressing topical areas in ocular surface physiology with new LFU proteins may reveal interesting new biological mechanisms and help get to the heart of ocular surface dysfunction. PMID:18840430

  1. Lacritin and other new proteins of the lacrimal functional unit.

    PubMed

    McKown, Robert L; Wang, Ningning; Raab, Ronald W; Karnati, Roy; Zhang, Yinghui; Williams, Patricia B; Laurie, Gordon W

    2009-05-01

    The lacrimal functional unit (LFU) is defined by the 2007 International Dry Eye WorkShop as 'an integrated system comprising the lacrimal glands, ocular surface (cornea, conjunctiva and meibomian glands) and lids, and the sensory and motor nerves that connect them'. The LFU maintains a healthy ocular surface primarily through a properly functioning tear film that provides protection, lubrication, and an environment for corneal epithelial cell renewal. LFU cells express thousands of proteins. Over 200 new LFU proteins have been discovered in the last decade. Lacritin is a new LFU-specific growth factor in human tears that flows through ducts to target corneal epithelial cells on the ocular surface. When applied topically in rabbits, lacritin appears to increase the volume of basal tear secretion. Lacritin is one of only a handful of tear proteins preliminarily reported to be downregulated in blepharitis and in two dry eye syndromes. Computational analysis predicts an ordered C-terminal domain that binds the corneal epithelial cell surface proteoglycan syndecan-1 (SDC1) and is required for lacritin's low nanomolar mitogenic activity. The lacritin-binding site on the N-terminus of SDC1 is exposed by heparanase. Heparanase is constitutively expressed by the corneal epithelium and appears to be a normal constituent of tears. Binding triggers rapid signaling to downstream NFAT and mTOR. A wealth of other new proteins, originally designated as hypothetical when first identified by genomic sequencing, are expressed by the human LFU including: ALS2CL, ARHGEF19, KIAA1109, PLXNA1, POLG, WIPI1 and ZMIZ2. Their demonstrated or implied roles in human genetic disease or basic cellular functions are fuel for new investigation. Addressing topical areas in ocular surface physiology with new LFU proteins may reveal interesting new biological mechanisms and help get to the heart of ocular surface dysfunction.

  2. {beta}-K{sub 4}La{sub 6}I{sub 14}Os: A new structure type for rare-earth-metal cluster compounds that contains discrete tetrahedral K{sub 4}I{sup 3+} units

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

    Uma, S.; Corbett, J.D.

    1999-08-23

    Suitable reactions of KI, La, LaI{sub 3}, and Os in niobium tubes at 800--850 C result in black, air- and moisture-sensitive crystals of the quaternary title phase. Isostructural K{sub 4}Pr{sub 6}I{sub 14}Z also exist for Z = Fe, Ru. The title phase was characterized by single-crystal X-ray diffraction (tetragonal, P4/ncc (No. 130), Z = 4; a = 13.117(3), c = 25.17(1) {angstrom} at 23 C). The important structural feature is the constitution (K{sub 4}I){sup 3+}(La{sub 6}I{sub 13}Os{sup 3{minus}}) with a new type of 3D anion network of [(La{sub 6}Os)I{sub 8}{sup i}I{sub 4/2}{sup i{minus}a}I{sub 4/2}{sup a{minus}i}I{sub 2/2}{sup a{minus}a}] clusters that aremore » connected into puckered layers through I{sup i{minus}a} and I{sup a{minus}i} atom pairs that bridge diagonally in the a-b plane. These cluster layers are further interlinked along {rvec c} at trans-vertexes through simple bridging I{sup a{minus}a}. The 14th iodine atom occurs in the unique K{sub 4}I{sup 3+} ions which lie in columns that interpenetrate the La{sub 6}OsI{sub 13} network along c. The present 16-e{sup {minus}} clusters, in contrast with the optimal 18-e{sup {minus}} octahedral cluster configuration, exhibit an uncommon tetragonal elongation and evidently become closed shell, with only a small temperature-independent (van Vleck-like) paramagnetism, {approximately}4 x 10{sup {minus}4} emu mol{sup {minus}1}.« less

  3. Magnetic, ferroelectric, and spin phonon coupling studies of Sr{sub 3}Co{sub 2}Fe{sub 24}O{sub 41} multiferroic Z-type hexaferrite

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

    Raju, N.; Shravan Kumar Reddy, S.; Ramesh, J.

    2016-08-07

    The magnetic, Raman, ferroelectric, and in-field {sup 57}Fe Mössbauer studies of polycrystalline multiferroic Sr{sub 3}Co{sub 2}Fe{sub 24}O{sub 41} are reported in this paper. From the magnetization studies, it is observed that the sample is soft magnetic in nature with low temperature magnetic spin transitions like longitudinal to transverse conical structure around 130 K and change in magnetic crystalline anisotropy from conical to planar structure at 250 K. Ferroelectric studies of the sample exhibit the spontaneous polarization at low temperature. Strong spin phonon and spin lattice coupling is observed through low temperature Raman spectroscopy. From the in-field {sup 57}Fe Mössbauer spectroscopy, spin upmore » and spin down site occupations of Fe ions are calculated in the unit cell.« less

  4. Synthesis, X-ray structure, and physical and photophysical properties of the heterobimetallic complex Fe({eta}{sup 5}-C{sub 5}H{sub 4}PPh{sub 2}){sub 2}Pt(bph)

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

    Zheng, G.Y.; Rillema, D.P.; Reibenspies, J.H.

    1999-02-22

    A chromophore-electroactive compound, Fe({eta}{sup 5}-C{sub 5}H{sub 4}PPh{sub 2}){sub 2}Pt(bph), where bph is the biphenyl dianion and Fe({eta}{sup 5}C{sub 5}H{sub 4}PPh{sub 2}){sub 2} is 1,1{prime}-bis(diphenylphosphino)ferrocene-P,P{prime} has been synthesized. The single-crystal X-ray structural characteristics of this heterobimetallic complex and its disolvated methylene chloride derivative are respectively as follows: empirical formula C{sub 46}H{sub 36}FeP{sub 2}Pt. An electrochemical study shows that the anodic potential for the oxidation of the ferrocenyl moiety of this compound increases by +0.13 V, compared to that for Fe({eta}{sup 5}-C{sub 5}H{sub 4}PPh{sub 2}){sub 2}. This change in oxidation potential agrees well with the change in energy of 0.11 eV formore » the d{pi}(Fe) {yields} {pi}{sup *}(Cp) MLCT transition upon coordination with Pt. The resultant excited state from the d{pi}(Pt) {yields} {pi}{sup *}(bph) MLCT transition is readily quenched by the ferrocenyl moiety unit as expected, and charge-separated redox-active centers are formed.« less

  5. Guanine nucleotide-binding regulatory proteins in retinal pigment epithelial cells

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

    Jiang, Meisheng; Tran, V.T.; Fong, H.K.W.

    1991-05-01

    The expression of GTP-binding regulatory proteins (G proteins) in retinal pigment epithelial (RPE) cells was analyzed by RNA blot hybridization and cDNA amplification. Both adult and fetal human RPE cells contain mRNA for multiple G protein {alpha} subunits (G{alpha}) including G{sub s}{alpha}, G{sub i-1}{alpha}, G{sub i-2}{alpha}, G{sub i-3}{alpha}, and G{sub z}{alpha} (or G{sub x}{alpha}), where G{sub s} and G{sub i} are proteins that stimulate or inhibit adenylyl cyclase, respectively, and G{sub z} is a protein that may mediate pertussis toxin-insensitive events. Other G{alpha}-related mRNA transcripts were detected in fetal RPE cells by low-stringency hybridization to G{sub i-2}{alpha} and G{sub s}{alpha}more » protein-coding cDNA probes. The diversity of G proteins in RPE cells was further studied by cDNA amplification with reverse transcriptase and the polymerase chain reaction. This approach revealed that, besides the above mentioned members of the G{alpha} gene family, at least two other G{alpha} subunits are expressed in RPE cells. Human retinal cDNA clones that encode one of the additional G{alpha} subunits were isolated and characterized. The results indicate that this G{alpha} subunit belongs to a separate subfamily of G proteins that may be insensitive to inhibition by pertussis toxin.« less

  6. Muscle protein turnover in rats treated with corticosterone (CC) or/and nandrolone decanoate (ND) and fed an adequate or a low-protein diet

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

    Santidrian, S.; Cuevillas, F.; Goena, M.

    1986-03-01

    In order to investigate the possible antagonistic effect between glucocorticoids and androgens on muscle protein turnover, the authors have measured the fractional rates of gastrocnemius muscle protein synthesis (k/sub s/) and degradation (k/sub d/) by the constant-intravenous-infusion method using L-//sup 14/C/-tyrosine in rats receiving via s.c. per 100 g b.wt. 10 mg of CC, or 2 mg of ND or CC+ND at the indicated doses, and fed either an 18% or 5% protein diets over a period of 5 days. As an additional index of protein synthesis, RNA activity (g of synthesized protein/day/g RNA) was determined as well. Results showedmore » that as compared to vehicle-injected animals fed the adequate diet, CC-treated rats exhibited a reduction of muscle k/sub d/, while ND-treated rats had an outstanding increase of muscle k/sub s/. However, rats receiving CC+ND showed k/sub s/ and k/sub d/ values similar to those displayed by control animals. Nevertheless, when the steroids were injected to rats fed the low-protein diet, CC has a catabolic effect on muscle protein but by reducing k/sub s/, while the anabolic action of ND is still displayed but by a significant reduction of muscle k/sub d/. CC+ND given to these protein-deficient rats caused an increase in muscle k/sub s/ and a reduction in k/sub d/. These results might indicate that, at least in part, ND antagonizes the catabolic action of high doses of CC on muscle protein metabolism.« less

  7. Crystal growth of incommensurate members of 2H-hexagonal perovskite related oxides: Ba{sub 4}M{sub z}Pt{sub 3−z}O{sub 9} (M=Co, Ni, Cu, Zn, Mg, Pt)

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

    Ferreira, Timothy; Morrison, Gregory; Yeon, Jeongho

    2016-04-15

    Millimeter sized crystals of six oxides of approximate composition Ba{sub 4}M{sub z}Pt{sub 3-z}O{sub 9} (M=Co, Ni, Cu, Zn, Mg, Pt) were grown from molten K{sub 2}CO{sub 3} fluxes and found to crystallize in a 2H hexagonal perovskite-related structure type. The compositions of these incommensurate structures, which belong to the A{sub 3n+3m}A′{sub n}B{sub 3m+n}O{sub 9m+6n} family of 2H hexagonal perovskite related oxides, were characterized by X-ray diffraction, energy dispersive spectroscopy, and magnetic susceptibility measurements. The specific synthetic considerations, crystal growth conditions, and magnetic susceptibility measurements are discussed. - Graphical abstract: SEM image and average commensurate unit cell of Ba{sub 4}Pt{sub 3}O{submore » 9.} - Highlights: • Single crystals of the series Ba{sub 4}A′{sub z}Pt{sub 3-z}O{sub 9} were grown via a molten carbonate flux. • Ba{sub 4}Pt{sub 3}O{sub 9} and all substitutional variants are incommensurate, composite structures. • All compounds have an approximate stoichiometry of Ba{sub 4}A′Pt{sub 2}O{sub 9.}.« less

  8. Enhancement of Gag-specific but reduction of Env- and Pol-specific CD8+ T cell responses by simian immunodeficiency virus nonstructural proteins in mice.

    PubMed

    Zhang, Yinfeng; Sun, Caijun; Feng, Liqiang; Xiao, Lijun; Chen, Ling

    2012-04-01

    Accessory and regulatory proteins (nonstructural proteins) have received increasing attention as components in novel HIV/SIV vaccine design. However, the complicated interactions between nonstructural proteins and structural proteins remain poorly understood, especially their effects on immunogenicity. In this study, the immunogenicity of structural proteins in the presence and absence of nonstructural proteins was compared. First, a series of recombinant plasmids and adenoviral vectors carrying various SIVmac239 nonstructural and structural genes was constructed. Then mice were primed with DNA plasmids and boosted with corresponding Ad5 vectors of different combinations, and the resulting immune responses were measured. Our results demonstrated that when the individual Gag, Pol, or Env gene products were coimmunized with the whole repertoire of nonstructural proteins, the Gag-specific CD8(+) T response was greatly enhanced, while the Env- and Pol-specific CD8(+) T responses were significantly reduced. The same pattern was not observed in CD4(+) T cell responses. Antibody responses against both the Gag and Env proteins were elicited more effectively when these structural antigens were immunized together with nonstructural antigens. These findings may provide helpful insights into the development of novel HIV/SIV vaccines.

  9. Pressure-induced amorphization of La{sub 1/3}TaO{sub 3}

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

    Noked, O., E-mail: noked@bgu.ac.il; Physics Department, Ben-Gurion University, Beer Sheva 84105; Melchior, A.

    2013-06-15

    La{sub 1/3}TaO{sub 3}, an A-site cation deficient perovskite, has been studied under pressure by synchrotron X-ray powder diffraction and Raman spectroscopy. It undergoes irreversible pressure induced amorphization at P=18.5 GPa. An almost linear unit cell volume decrease vs. pressure is observed from ambient pressure up to the phase transition. The Raman spectroscopy also shows amorphization at the same pressure, with positive shifts of all modes as a function of pressure. The pressure dependence of the E{sub g} and A{sub 1g} Raman modes arising from the octahedral oxygen network is discussed. - Graphical abstract: La{sub 1/3}Tao{sub 3} exhibits linear pressure–volume relationmore » until irreversible pressure induced amorphization at 18.5 Gpa. - Highlights: • La{sub 1/3}TaO{sub 3} has been studied under pressure by synchrotron XRD and Raman spectroscopy. • La{sub 1/3}TaO{sub 3} undergoes irreversible pressure induced amorphization around 18.5 GPa. • The transition is manifested in both XRD and Raman measurements. • A linear P–V relation is observed from ambient pressure up to the phase transition.« less

  10. A new anion-deficient fluorite-related superstructure of Bi{sub 28}V{sub 8}O{sub 62}

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

    Đorđević, T., E-mail: tamara.djordjevic@univie.ac.at; Karanović, Lj., E-mail: ljika2002@yahoo.com

    2014-12-15

    New hydrothermally synthesized Bi{sub 28}V{sub 8}O{sub 62} was structurally characterized using single-crystal X-ray diffraction data. Bi{sub 28}V{sub 8}O{sub 62} crystallizes in the novel type of defect fluorite structure related to the face-centered cubic δ-Bi{sub 2}O{sub 3}. It is monoclinic, s. g. P2{sub 1}/c, and the relation to the fluorite subcell is given as a∼(3/2)a{sub F}+(3/2)c{sub F}; b∼ −b{sub F}; c∼2a{sub F} −4c{sub F} (F in subscript indicate the unit cell parameter of fluorite). Its structure is characterized by slabs of edge sharing OBi{sub 4} tetrahedra surrounded by the OBi{sub 3} triangles. As a part of these OBi{sub 3} triangles, twomore » positionally disordered Bi{sup 3+} cations were observed in the marginal part of the slabs. The slabs are extending along b axis and are linked by inter-slab portion of the structure composed of VO{sub 4} tetrahedra and BiO{sub 6−x} coordination polyhedra, where x is a number of vacant oxygen sites. Raman spectra verified the coordination environment of vanadium atoms in the structure. - Graphical abstract: The [4{sup ¯}01] projection of two slabs and inter-slab part of the structure in one layer parallel to the (3{sup ¯}08)=(002{sup ¯}){sub F} plane (F in subscript indicate a fluorite type structure). The large green circles are Bi atoms. Small blue circles represent partly and fully occupied O sites, respectively. Pink (hatched black) are V1O{sub 4} and blue (hatched white) are V2O{sub 4} coordination tetrahedra. - Highlights: • Single crystals of Bi{sub 28}V{sub 8}O{sub 62} were grown using hydrothermal technique. • The crystal structure of Bi{sub 28}V{sub 8}O{sub 62} was solved using single-crystal XRD method. • Bi{sub 28}V{sub 8}O{sub 62} has an anion-deficient fluorite-related superstructure. • Raman spectrum confirmed the coordination environment of vanadium atoms. • Relation to the structurally related compound was discussed.« less

  11. Hyperoside attenuates hydrogen peroxide-induced L02 cell damage via MAPK-dependent Keap{sub 1}-Nrf{sub 2}-ARE signaling pathway

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

    Xing, Hai-Yan; Liu, Yao; Chen, Jian-Hong

    Highlights: {yields} Hyperoside attenuated H{sub 2}O{sub 2}-induced L02 cell damage. {yields} Hyperoside up-regulated HO-1 expression at both mRNA and protein levels. {yields} Hyperoside activated both Nrf{sub 2} nuclear translocation and gene expression. {yields} Hyperoside may inhibit Keap{sub 1} mRNA translation or protein degradation. {yields} Phosphorylation of ERK and p38 is involved in hyperoside-mediated Nrf{sub 2} activation. -- Abstract: The flavonoid hyperoside has been reported to elicit cytoprotection against oxidative stress partly by increasing the activity of antioxidant enzymes, such as glutathione peroxidase, superoxide dismutase and catalase. However, the cellular and molecular mechanisms underlying this effect remain unclear. Here, hepatic L02more » cells exposed to H{sub 2}O{sub 2} (100 {mu}M) were used to demonstrate that hyperoside protected cells by significantly inhibiting overproduction of intracellular ROS, depletion of the mitochondrial membrane potential and leakage of lactate dehydrogenase. Hyperoside further enhanced the cellular antioxidant defense system through increasing the activity of heme oxygenase-1 (HO-1), and by up-regulating HO-1 expression. Meanwhile, real time PCR, western blot and immunofluorescence studies revealed that hyperoside stimulated nuclear translocation of the Nrf{sub 2} transcription factor in a dose-dependent manner, and this effect was significantly suppressed by pharmacological inhibition of the mitogen-activated protein kinases (MAPK) p38 and ERK. Collectively, our data provide the first description of the mechanism underlying hyperoside's ability to attenuate H{sub 2}O{sub 2}-induced cell damage, namely this compound interacts with the MAPK-dependent Keap{sub 1}-Nrf{sub 2}-ARE signaling pathway to up-regulate HO-1 expression and enhance intracellular antioxidant activity.« less

  12. High-pressure/high-temperature synthesis and characterization of the first palladium or platinum containing lithium transition-metal sulfides Li{sub 2}M{sub 3}S{sub 4} (M=Pd, Pt)

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

    Heymann, Gunter, E-mail: Gunter.Heymann@uibk.ac.at; Niehaus, Oliver; Krüger, Hannes

    The new lithium transition-metal sulfides Li{sub 2}M{sub 3}S{sub 4} (M=Pd, Pt) were obtained via multianvil high-pressure/high-temperature syntheses at 8 GPa and 1150 °C starting from a stoichiometric mixture of lithium nitride, sulfur, and palladium or platinum. Single crystal structure analyses indicated the space group P2{sub 1}/c (no. 14) with the following lattice parameters and refinement results: a=492.9(1), b=1005.9(2), c=614.9(2) pm, β=110.9 (1)°, R1=0.0165, wR2=0.0308 (all data) for Li{sub 2}Pd{sub 3}S{sub 4} and a=498.2(1), b=1005.5(2), c=613.0(2) pm, β=110.8(1)°, R1=0.0215, wR2=0.0450 (all data) for Li{sub 2}Pt{sub 3}S{sub 4}. The crystal structures are built up from two distinct Pd/Pt sites, one of whichmore » is a special position (0,0,0), two sulfur sites, and one lithium site. The atoms Pd2/Pt2 form isolated square planar PdS{sub 4}/PtS{sub 4} units, whereas the Pd1/Pt1 atoms form pairs of square planar PdS{sub 4}/PtS{sub 4} units, which are connected via a common edge. These two structural motives built up a three-dimensional network structure by linking through common corners. The lithium atoms are positioned inside of the so formed channels. Li{sub 2}M{sub 3}S{sub 4} (M=Pd, Pt) are isostructural to the minerals jaguéite, Cu{sub 2}Pd{sub 3}Se{sub 4} and chrisstanleyite, Ag{sub 2}Pd{sub 3}Se{sub 4}, which are up to now the only representatives of this structure type. Both compounds were studied with respect to their magnetic properties and can be classified as Pauli paramagnetic or diamagnetic. Regarding the possibility of lithium mobility inside the channels, of the structure, solid state {sup 7}Li NMR and high-temperature single crystal investigations revealed localization of the lithium atoms on their crystallographic sites. - Graphical abstract: The ternary lithium transition-metal sulfides Li{sub 2}M{sub 3}S{sub 4} (M=Pd, Pt) were prepared via multianvil high-pressure/high-temperature syntheses. They are built up from square planar PtS{sub

  13. The cluster compound In{sub 4}Ti{sub 1.5}Mo{sub 0.5}Mo{sub 14}O{sub 26} containing Mo{sub 14} clusters and the new mono- and bi-capped trioctahedral Mo{sub 15} and Mo{sub 16} clusters: Synthesis, crystal structure, and electrical and magnetic properties

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

    Gall, Philippe; Guizouarn, Thierry; Gougeon, Patrick, E-mail: Patrick.Gougeon@univ-rennes1.fr

    2015-07-15

    Single crystals of the new quaternary compound In{sub 4}Ti{sub 1.5}Mo{sub 0.5}Mo{sub 14}O{sub 26} were obtained by solid state reaction. The crystal structure was determined by single-crystal X-ray diffraction. In{sub 4}Ti{sub 1.5}Mo{sub 0.5}Mo{sub 14}O{sub 26} crystallizes in the orthorhombic space group Pbca with unit-cell parameters a=9.4432(14) Å, b=11.4828(12) Å, c=20.299(4) Å and Z=4. Full-matrix least-squares refinement on F{sup 2} using 3807 independent reflections for 219 refinable parameters resulted in R{sub 1}=0.0259 and wR{sub 2}=0.0591. The crystal structure contains in addition to Mo{sub 14} clusters the first examples of mono- and bi-capped trioctahedral Mo{sub 14} i.e. Mo{sub 15} and Mo{sub 16} clusters.more » The oxygen framework derives from a stacking along the a direction of close-packed layers with sequence (…ABAC…). The Mo–Mo distances range between 2.6938(5) and 2.8420(6) Å and the Mo–O distances between 1.879(5) and 2.250(3) Å, as usually observed in molybdenum oxide clusters. The indium atoms form In{sub 4}{sup 6+} bent chains with In–In distances of 2.6682(5) and 2.6622(8) Å and the Ti atoms are in highly distorted octahedral sites of oxygen atoms with Ti–O distances ranging between 1.865(4) and 2.161(4) Å. Magnetic susceptibility measurements confirm the presence of Ti{sup 4+} cations and the absence of localized moments on the Mo network. Electrical resistivity measurements on a single crystal of In{sub 4}Ti{sub 1.5}Mo{sub 0.5}Mo{sub 14}O{sub 26} show a semimetallic behavior. - Graphical abstract: We present here the synthesis, the crystal structure, and the electrical and magnetic properties of the new compound In{sub 4}Ti{sub 1.5}Mo{sub 0.5}Mo{sub 14}O{sub 26} in which Mo{sub 14} clusters coexist statistically with mono- and bi-capped trioctahedral Mo{sub 14} that is Mo{sub 15} and Mo{sub 16} clusters. - Highlights: • Single crystals of In{sub 4}Ti{sub 1.5}Mo{sub 0.5}Mo{sub 14}O{sub 26} were obtained by solid

  14. High-pressure synthesis and crystal structures of the strontium oxogallates Sr{sub 2}Ga{sub 2}O{sub 5} and Sr{sub 5}Ga{sub 6}O{sub 14}

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

    Kahlenberg, Volker, E-mail: volker.kahlenberg@uibk.ac.at; Goettgens, Valerie; Mair, Philipp

    2015-08-15

    High-pressure synthesis experiments in a piston–cylinder apparatus at 1.5 GPa/3.0 GPa and 1000 °C resulted in the formation of single-crystals of Sr{sub 2}Ga{sub 2}O{sub 5} and Sr{sub 5}Ga{sub 6}O{sub 14}, respectively. The structures of both compounds have been solved from single-crystal diffraction data sets using direct methods. The first compound is orthorhombic with space group type Pbca (a=10.0021(4) Å, b=9.601(4) Å, c=10.6700(4) Å, V=1024.6(4) Å{sup 3}, M{sub r}=394.68 u, Z=8, D{sub x}=5.12 g/cm{sup 3}) and belongs to the group of single layer gallates. Individual sheets are parallel to (0 0 1) and can be built from the condensation of unbranchedmore » vierer single chains running along [0 1 0]. The layers are characterized by the presence of four- and strongly elliptical eight-membered rings of corner connected tetrahedra in UUDD and UUUUDDDD conformation. Strontium atoms are sandwiched between the tetrahedral layers for charge compensation and are coordinated by six and seven oxygen ligands, respectively. Sr{sub 2}Ga{sub 2}O{sub 5} is isotypic with several other double sulfides and selenides. To the best of our knowledge, it is the first example of an oxide with this structure type. From a structural point of view, Sr{sub 5}Ga{sub 6}O{sub 14} is a phyllogallate as well. The crystal structure adopts the monoclinic space group P2{sub 1}/c (a=8.1426(3) Å, b=8.1803(3) Å, c=10.8755(4) Å, β=91.970(4)° V=723.98(5) Å{sup 3}, M{sub r}=1080.42 u, Z=2, D{sub x}=4.96 g/cm{sup 3}). Individual sheets extend along (0 0 1). Basic building units are unbranched dreier single chains parallel to [1 0 0]. The layers contain tertiary (Q{sup 3}) und quaternary (Q{sup 4}) connected [GaO{sub 4}]-tetrahedra in the ratio 2:1 resulting in a Ga:O ratio of 3:7 and the formation of exclusively five-membered rings. Linkage between adjacent tetrahedral sheets is provided by three symmetrically independent strontium ions which are surrounded by six to eight oxygen atoms. The

  15. Crystallization and preliminary crystallographic analysis of calcium-binding protein-2 from Entamoeba histolytica and its complexes with strontium and the IQ1 motif of myosin V

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

    Gourinath, S., E-mail: sgourinath@mail.jnu.ac.in; Padhan, Narendra; Alam, Neelima

    2005-04-01

    Calcium-binding protein-2 (EhCaBP2) crystals were grown using MPD as a precipitant. EhCaBP2 also crystallized in complex with strontium (replacing calcium) at similar conditions. Preliminary data for EhCaBP2 crystals in complex with an IQ motif are also reported. Calcium plays a pivotal role in the pathogenesis of amoebiasis, a major disease caused by Entamoeba histolytica. Two domains with four canonical EF-hand-containing calcium-binding proteins (CaBPs) have been identified from E. histolytica. Even though they have very high sequence similarity, these bind to different target proteins in a Ca{sup 2+}-dependent manner, leading to different functional pathways. Calcium-binding protein-2 (EhCaBP2) crystals were grown usingmore » MPD as a precipitant. The crystals belong to space group P2{sub 1}, with unit-cell parameters a = 111.74, b = 68.83, c = 113.25 Å, β = 116.7°. EhCaBP2 also crystallized in complex with strontium (replacing calcium) at similar conditions. The crystals belong to space group P2{sub 1}, with unit-cell parameters a = 69.18, b = 112.03, c = 93.42 Å, β = 92.8°. Preliminary data for EhCaBP2 crystals in complex with an IQ motif are also reported. This complex was crystallized with MPD and ethanol as precipitating agents. These crystals belong to space group P2{sub 1}, with unit-cell parameters a = 60.5, b = 69.86, c = 86.5 Å, β = 97.9°.« less

  16. Protein Conformational Populations and Functionally Relevant Sub-states

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

    Agarwal, Pratul K; Burger, Virginia; Savol, Andrej

    2013-01-01

    Functioning proteins do not remain fixed in a unique structure, but instead they sample a range of conformations facilitated by motions within the protein. Even in the native state, a protein exists as a collection of interconverting conformations driven by thermodynamic fluctuations. Motions on the fast time scale allow a protein to sample conformations in the nearby area of its conformational landscape, while motions on slower time scales give it access to conformations in distal areas of the landscape. Emerging evidence indicates that protein landscapes contain conformational substates with dynamic and structural features that support the designated function of themore » protein. Nuclear magnetic resonance (NMR) experiments provide information about conformational ensembles of proteins. X-ray crystallography allows researchers to identify the most populated states along the landscape, and computational simulations give atom-level information about the conformational substates of different proteins. This ability to characterize and obtain quantitative information about the conformational substates and the populations of proteins within them is allowing researchers to better understand the relationship between protein structure and dynamics and the mechanisms of protein function. In this Account, we discuss recent developments and challenges in the characterization of functionally relevant conformational populations and substates of proteins. In some enzymes, the sampling of functionally relevant conformational substates is connected to promoting the overall mechanism of catalysis. For example, the conformational landscape of the enzyme dihydrofolate reductase has multiple substates, which facilitate the binding and the release of the cofactor and substrate and catalyze the hydride transfer. For the enzyme cyclophilin A, computational simulations reveal that the long time scale conformational fluctuations enable the enzyme to access conformational substates that

  17. [La(UO{sub 2})V{sub 2}O{sub 7}][(UO{sub 2})(VO{sub 4})] the first lanthanum uranyl-vanadate with structure built from two types of sheets based upon the uranophane anion-topology

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

    Mer, A.; Obbade, S.; Rivenet, M.

    2012-01-15

    The new lanthanum uranyl vanadate divanadate, [La(UO{sub 2})V{sub 2}O{sub 7}][(UO{sub 2})(VO{sub 4})] was obtained by reaction at 800 Degree-Sign C between lanthanum chloride, uranium oxide (U{sub 3}O{sub 8}) and vanadium oxide (V{sub 2}O{sub 5}) and the structure was determined from single-crystal X-ray diffraction data. This compound crystallizes in the orthorhombic system with space group P2{sub 1}2{sub 1}2{sub 1} and unit-cell parameters a=6.9470(2) A, b=7.0934(2) A, c=25.7464(6) A, V=1268.73(5) A{sup 3}, Z=4. A full matrix least-squares refinement yielded R{sub 1}=0.0219 for 5493 independent reflections. The crystal structure is characterized by the stacking of uranophane-type sheets {sup 2}{sub {infinity}}[(UO{sub 2})(VO{sub 4})]{sup -}more » and double layers {sup 2}{sub {infinity}}[La(UO{sub 2})(V{sub 2}O{sub 7})]{sup +} connected through La-O bonds involving the uranyl oxygen of the uranyl-vanadate sheets. The double layers result from the connection of two {sup 2}{sub {infinity}}[La(UO{sub 2})(VO{sub 4}){sub 2}]{sup -} sheets derived from the uranophane anion-topology by replacing half of the uranyl ions by lanthanum atoms and connected through the formation of divanadate entities. - Graphical abstract: A view of the three-dimensional structure of [La(UO{sub 2})V{sub 2}O{sub 7}][(UO{sub 2})(VO{sub 4})]. Highlights: Black-Right-Pointing-Pointer New lanthanum uranyl vanadate divanadate has been synthesized. Black-Right-Pointing-Pointer Structure was determined from single-crystal X-ray diffraction data. Black-Right-Pointing-Pointer Structure is characterized by uranophane-type sheets and double layers {sup 2}{sub {infinity}}[La(UO{sub 2})(V{sub 2}O{sub 7})]{sup +}.« less

  18. Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of macrophage growth locus A (MglA) protein from Francisella tularensis

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

    Subburaman, P.; Austin, B.P.; Shaw, G.X.

    2010-11-03

    Francisella tularensis, a potential bioweapon, causes a rare infectious disease called tularemia in humans and animals. The macrophage growth locus A (MglA) protein from F. tularensis associates with RNA polymerase to positively regulate the expression of multiple virulence factors that are required for its survival and replication within macrophages. The MglA protein was overproduced in Escherichia coli, purified and crystallized. The crystals diffracted to 7.5 {angstrom} resolution at the Advanced Photon Source, Argonne National Laboratory and belonged to the hexagonal space group P6{sub 1} or P6{sub 5}, with unit-cell parameters a = b = 125, c = 54 {angstrom}.

  19. China's Pathways to Achieving 40% ~ 45% Reduction in CO{sub 2} Emissions per Unit of GDP in 2020: Sectoral Outlook and Assessment of Savings Potential

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

    Zheng, Nina; Fridley, David; Zhou, Nan

    2011-09-30

    Achieving China’s goal of reducing its carbon intensity (CO{sub 2} per unit of GDP) by 40% to 45% percent below 2005 levels by 2020 will require the strengthening and expansion of energy efficiency policies across the buildings, industries and transport sectors. This study uses a bottom-up, end-use model and two scenarios -- an enhanced energy efficiency (E3) scenario and an alternative maximum technically feasible energy efficiency improvement (Max Tech) scenario – to evaluate what policies and technical improvements are needed to achieve the 2020 carbon intensity reduction target. The findings from this study show that a determined approach by Chinamore » can lead to the achievement of its 2020 goal. In particular, with full success in deepening its energy efficiency policies and programs but following the same general approach used during the 11th Five Year Plan, it is possible to achieve 49% reduction in CO{sub 2} emissions per unit of GDP (CO{sub 2} emissions intensity) in 2020 from 2005 levels (E3 case). Under the more optimistic but feasible assumptions of development and penetration of advanced energy efficiency technology (Max Tech case), China could achieve a 56% reduction in CO{sub 2} emissions intensity in 2020 relative to 2005 with cumulative reduction of energy use by 2700 Mtce and of CO{sub 2} emissions of 8107 Mt CO{sub 2} between 2010 and 2020. Energy savings and CO{sub 2} mitigation potential varies by sector but most of the energy savings potential is found in energy-intensive industry. At the same time, electricity savings and the associated emissions reduction are magnified by increasing renewable generation and improving coal generation efficiency, underscoring the dual importance of end-use efficiency improvements and power sector decarbonization.« less

  20. Protein crystal growth results from the United States Microgravity Laboratory-1 mission

    NASA Technical Reports Server (NTRS)

    Delucas, Lawrence J.; Moore, K. M.; Vanderwoerd, M.; Bray, T. L.; Smith, C.; Carson, M.; Narayana, S. V. L.; Rosenblum, W. M.; Carter, D.; Clark, A. D, Jr.

    1994-01-01

    Protein crystal growth experiments have been performed by this laboratory on 18 Space Shuttle missions since April, 1985. In addition, a number of microgravity experiments also have been performed and reported by other investigators. These Space Shuttle missions have been used to grow crystals of a variety of proteins using vapor diffusion, liquid diffusion, and temperature-induced crystallization techniques. The United States Microgravity Laboratory - 1 mission (USML-1, June 25 - July 9, 1992) was a Spacelab mission dedicated to experiments involved in materials processing. New protein crystal growth hardware was developed to allow in orbit examination of initial crystal growth results, the knowledge from which was used on subsequent days to prepare new crystal growth experiments. In addition, new seeding hardware and techniques were tested as well as techniques that would prepare crystals for analysis by x-ray diffraction, a capability projected for the planned Space Station. Hardware that was specifically developed for the USML-1 mission will be discussed along with the experimental results from this mission.

  1. Synthesis and characterization of two crystallographic forms of Ag{sub 0.79}VS{sub 2}

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

    Ali, Mazhar N., E-mail: maz@berkeley.edu; Ji, Huiwen; Hirai, Daigorou

    2013-06-15

    A previously unreported compound, Ag{sub 0.79}VS{sub 2}, has been synthesized; its structure and elementary properties are reported. Ag{sub 0.79}VS{sub 2} crystallizes in two forms, designated as the α and β, related to the 1s-InTaS{sub 2} structure. Single crystal x-ray diffraction shows the α form to have a single layer hexagonal structure with a unit cell of 3.213(3) Å×7.809(6) Å, consisting of layers of edge-shared VS{sub 6} triangular prisms separated by layers of Ag. The β form is similar but has an a{sub o}√((3)) supercell in the basal plane, yielding a unit cell of 5.573(5) Å×7.822(6) Å. Both forms have disorderedmore » and displaced silver in the basal plane, but the β form has partial ordering of its silver sublattice and in-plane vanadium trimers. Resistivity measurements show metallic temperature dependence with an unusual hysteresis between 210 K and 130 K. Magnetic susceptibility measurements show Pauli Paramagnetic behavior. The Seebeck coefficient at 300 K is 42 µV/K. - Graphical abstract: Red=Vanadium, Gray=Silver, Yellow=Sulfur. Top left is α-Ag{sub 0.79}VS{sub 2} in the 1s-InTaS{sub 2} structure type. Top right: 2a×2b projection down the c-axis with displacement ellipsoids (50% probability) of atoms drawn to illustrate the split silver model. Bottom left is β-Ag{sub 0.79}VS{sub 2} having the a{sub o}√((3)) supercell. Bottom right: projection along the c-axis, displacement ellipsoids of atoms drawn. - Highlights: • Two crystallographic forms of Ag{sub 0.79}VS{sub 2}, designated α and β, are reported. • They are found to crystallize in the 1s-InTaS{sub 2} structure type. • β-Ag{sub 0.79}VS{sub 2} has partial ordering of the Ag sublattice and vanadium trimers. • Ag{sub 0.79}VS{sub 2} shows Pauli Paramagnetic behavior, a Seebeck coefficient of 42 µV/K at 300 K. • Ag{sub 0.79}VS{sub 2} shows metallic resistivity with an unusual hysteresis between 210 K and 130 K.« less

  2. Electric field induced cubic to monoclinic phase transition in multiferroic 0.65Bi(Ni{sub 1/2}Ti{sub 1/2})O{sub 3}-0.35PbTiO{sub 3} solid solution

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

    Pandey, Rishikesh; Singh, Akhilesh Kumar, E-mail: akhilesh-bhu@yahoo.com

    2014-10-20

    The results of x-ray diffraction studies on 0.65Bi(Ni{sub 1/2}Ti{sub 1/2})O{sub 3}-0.35PbTiO{sub 3} solid solution poled at various electric fields are presented. After poling, significant value of planar electromechanical coupling coefficient (k{sub P}) is observed for this composition having cubic structure in unpoled state. The cubic structure of 0.65Bi(Ni{sub 1/2}Ti{sub 1/2})O{sub 3}-0.35PbTiO{sub 3} transforms to monoclinic structure with space group Pm for the poling field ≥5 kV/cm. Large c-axis microscopic lattice strain (1.6%) is achieved at 30 kV/cm poling field. The variation of the c-axis strain and unit cell volume with poling field shows a drastic jump similar to that observed for strainmore » versus electric field curve in (1 − x)Pb(Mg{sub 1/3}Nb{sub 2/3}) O{sub 3}-xPbTiO{sub 3} and (1 − x)Pb(Zn{sub 1/3}Nb{sub 2/3})O{sub 3}-xPbTiO{sub 3}.« less

  3. A single amino acid substitution within the transmembrane domain of the human immunodeficiency virus type 1 Vpu protein renders simian-human immunodeficiency virus (SHIV{sub KU-1bMC33}) susceptible to rimantadine

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

    Hout, David R.; Gomez, Lisa M.; Pacyniak, Erik

    2006-05-10

    Previous studies from our laboratory have shown that the transmembrane domain (TM) of the Vpu protein of human immunodeficiency virus type 1 (HIV-1) contributes to the pathogenesis of SHIV{sub KU-1bMC33} in macaques and that the TM domain of Vpu could be replaced with the M2 protein viroporin from influenza A virus. Recently, we showed that the replacement of the TM domain of Vpu with that of the M2 protein of influenza A virus resulted in a virus (SHIV{sub M2}) that was sensitive to rimantadine [Hout, D.R., Gomez, M.L., Pacyniak, E., Gomez, L.M., Inbody, S.H., Mulcahy, E.R., Culley, N., Pinson, D.M.,more » Powers, M.F., Wong, S.W., Stephens, E.B., 2006. Substitution of the transmembrane domain of Vpu in simian human immunodeficiency virus (SHIV{sub KU-1bMC33}) with that of M2 of influenza A results in a virus that is sensitive to inhibitors of the M2 ion channel and is pathogenic for pig-tailed macaques. Virology 344, 541-558]. Based on previous studies of the M2 protein which have shown that the His-X-X-X-Trp motif within the M2 is essential to the function of the M2 proton channel, we have constructed a novel SHIV in which the alanine at position 19 of the TM domain was replaced with a histidine residue resulting in the motif His-Ile-Leu-Val-Trp. The SHIV{sub VpuA19H} replicated with similar kinetics as the parental SHIV{sub KU-1bMC33} and pulse-chase analysis revealed that the processing of viral proteins was similar to SHIV{sub KU-1bMC33}. This SHIV{sub VpuA19H} virus was found to be more sensitive to the M2 ion channel blocker rimantadine than SHIV{sub M2}. Electron microscopic examination of SHIV{sub VpuA19H}-infected cells treated with rimantadine revealed an accumulation of viral particles at the cell surface and within intracellular vesicles, which was similar to that previously observed to SHIV{sub M2}-infected cells treated with rimantadine. These data indicate that the Vpu protein of HIV-1 can be converted into a rimantadine-sensitive ion channel

  4. Synthesis and structure of novel lithium-ion conductor Li{sub 7}Ge{sub 3}PS{sub 12}

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

    Inoue, Yuki; Suzuki, Kota; Department of Chemical Science and Engineering, School of Materials and Chemical Technology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori, Yokohama 226-8502

    The novel lithium-ion conductor Li{sub 7}Ge{sub 3}PS{sub 12} was synthesized by slow cooling from the ternary Li{sub 2}S–GeS{sub 2}–P{sub 2}S{sub 5} system, and was shown to exhibit a cubic argyrodite-type structure. The phase composition was determined by varying the ratio of starting materials; the observed monophasic properties were close to those for the Li{sub 7}Ge{sub 3}PS{sub 12} composition. The lattice parameter (a =9.80192(3) Å) of Li{sub 7}Ge{sub 3}PS{sub 12} was slightly smaller than that of Li{sub 7}PS{sub 6} (a =9.993 Å), indicating that substitution of a Li cation by the smaller Ge cation contracted the cubic lattice. In addition, themore » novel structure consisted of a framework composed of four isolated (Ge/P)S{sub 4} tetrahedra. Li{sup +} ions occupied tetrahedral sites within the framework, forming a three-dimensional conduction pathway. Finally, Li{sub 7}Ge{sub 3}PS{sub 12} exhibited a high ionic conductivity of 1.1×10{sup −4} S cm{sup −1} at 25 °C and an activation energy of 25 kJ mol{sup −1}. - Graphical abstract: A novel Li{sub 7}Ge{sub 3}PS{sub 12} solid lithium ion conductor, with cubic argyrodite strucuture, shows high ion conductivity of 1.1×10{sup –4} S cm{sup –1} with an activation energy of 25 kJ mol{sup –1}. The argyrodite structure consists of (Ge/P)S{sub 4} tetrahedra units along with partial occupation of lithium and germanium at 48 h site. - Highlights: • A novel lithium-ion conductor Li{sub 7}Ge{sub 3}PS{sub 12} was detected. • This was achieved through slow cooling of the ternary Li{sub 2}S–GeS{sub 2}–P{sub 2}S{sub 5} system. • This novel conductor revealed a cubic argyrodite-type structure. • Li{sub 7}Ge{sub 3}PS{sub 12} exhibited a high ionic conductivity of 1.1×10{sup −4} S cm{sup −1} at 25 °C. • These properties will aid in the design of superior lithium-ion conductors.« less

  5. Preferential orientation relationships in Ca{sub 2}MnO{sub 4} Ruddlesden-Popper thin films

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

    Lacotte, M.; David, A.; Prellier, W., E-mail: wilfrid.prellier@ensicaen.fr

    2015-07-28

    A high-throughput investigation of local epitaxy (called combinatorial substrate epitaxy) was carried out on Ca{sub 2}MnO{sub 4} Ruddlesden-Popper thin films of six thicknesses (from 20 to 400 nm), all deposited on isostructural polycrystalline Sr{sub 2}TiO{sub 4} substrates. Electron backscatter diffraction revealed grain-over-grain local epitaxial growth for all films, resulting in a single orientation relationship (OR) for each substrate-film grain pair. Two preferred epitaxial ORs accounted for more than 90% of all ORs on 300 different microcrystals, based on analyzing 50 grain pairs for each thickness. The unit cell over unit cell OR ([100][001]{sub film} ∥ [100][001]{sub substrate}, or OR1) accounted formore » approximately 30% of each film. The OR that accounted for 60% of each film ([100][001]{sub film} ∥ [100][010]{sub substrate}, or OR2) corresponds to a rotation from OR1 by 90° about the a-axis. OR2 is strongly favored for substrate orientations in the center of the stereographic triangle, and OR1 is observed for orientations very close to (001) or to those near the edge connecting (100) and (110). While OR1 should be lower in energy, the majority observation of OR2 implies kinetic hindrances decrease the frequency of OR1. Persistent grain over grain growth and the absence of variations of the OR frequencies with thickness implies that the growth competition is finished within the first few nm, and local epitaxy persists thereafter during growth.« less

  6. Nuclear translocation of the 1,25D{sub 3}-MARRS (membrane associated rapid response to steroids) receptor protein and NF{kappa}B in differentiating NB4 leukemia cells

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

    Wu, Wenqing; Beilhartz, Greg; Roy, Yvette

    2010-04-15

    1,25 Dihydroxyvitamin D{sub 3} (1,25D{sub 3}) primes NB4 promyelocytic leukemia cells to differentiate along the monocyte/macrophage lineage through a non-genomic mechanism. Here we show that NB4 cells express high levels of the recently identified membrane receptor for 1,25D{sub 3}, which is a distinct gene product from the classical nuclear vitamin D receptor. This 57 kDa protein, named 1,25D{sub 3}-MARRS (Membrane Activated Rapid Response to Steroids)/ERp57/PIA3 appears to associate in a complex with the transcription factor, nuclear factor kappa B (NF{kappa}B). In unstimulated cells, 1,25D{sub 3}-MARRS can be co-immunoprecipitated with antibodies directed at NF{kappa}B, and NF{kappa}B is co-precipitated when antibodies againstmore » 1,25D{sub 3}-MARRS or ERp57 are used. Confocal microscopy and subcellular fractionation studies demonstrate that both 1,25D{sub 3}-MARRS and NF{kappa}B begin translocating to the nucleus within minutes of co-stimulation with 1,25D{sub 3} and phorbol ester. The predominant nuclear localization of both proteins precedes the expression of the monocyte/macrophage phenotype and suggests that this event may be critical to the differentiation pathway. This suggests a role for 1,25D{sub 3}-MARRS in the nucleus as a regulator of gene expression. Here it may also regulate the activity of NF{kappa}B and other factors with which it may be interacting.« less

  7. Phase transition, crystal water and low thermal expansion behavior of Al{sub 2−2x}(ZrMg){sub x}W{sub 3}O{sub 12}·n(H{sub 2}O)

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

    Li, Fang; Liu, Xiansheng; Song, Wenbo

    2014-10-15

    Al{sub 2−2x}(ZrMg){sub x}W{sub 3}O{sub 12} for 0≤x≤1.0 are synthesized to reduce the phase transition temperature of Al{sub 2}W{sub 3}O{sub 12}. It is found that the incorporation of (ZrMg){sup 6+} into the lattice of Al{sub 2}W{sub 3}O{sub 12} not only reduces its orthorhombic-to-monoclinic phase transition temperature but also elevates its softening temperature, broadening its applicable temperature range considerably. Al{sub 2−2x}(ZrMg){sub x}W{sub 3}O{sub 12} with x<0.5 exhibit low coefficients of thermal expansion (CTEs) and non-hygroscopicity, while those for x≥0.7 are obviously hygroscopic and the CETs decrease with increasing the content of (ZrMg){sup 6+} so that Al{sub 0.2}(ZrMg){sub 0.9}W{sub 3}O{sub 12} and ZrMgW{submore » 3}O{sub 12} exhibit negative thermal expansion. Temperature-dependent Raman spectroscopic study shows the hardening of W–O bonds above 373 K which is attributed to the release of crystal water. The effect of crystal water on the thermal expansion property is discussed based on the hydrogen bond between H in crystal water and electronegative O in Al(ZrMg)–O–W linkages. - Graphical abstract: (a and b) Temperature dependent Raman spectra of Al{sub 2−x}(ZrMg){sub x}W{sub 3}O{sub 12} (x=0.1, 0.2), (c and d) Building block of a unit cell of Al{sub 2−x}(ZrMg){sub x}W{sub 3}O{sub 12}·n(H{sub 2}O) and schematic showing the effect of crystal water on Al(Zr, Mg)–O–W linkages. - Highlights: • (ZrMg){sup 6+} reduces orthorhombic-to-monoclinic phase transition of Al{sub 2}W{sub 3}O{sub 12}. • The incorporation of (ZrMg){sup 6+} elevates the softening temperature of Al{sub 2}W{sub 3}O{sub 12}. • Al{sub 2−2x}(ZrMg){sub x}W{sub 3}O{sub 12} (x<0.5) exhibit low CTEs and non-hygroscopicity. • Al{sub 0.2}(ZrMg){sub 0.9}W{sub 3}O{sub 12}·0.8H{sub 2}O and ZrMgW{sub 3}O{sub 12}·2H{sub 2}O present NTE. • Hydrogen bond between H in H{sub 2}O and O in Al(ZrMg)–O–W affects thermal expansion.« less

  8. Atomistic study on the site preference and lattice vibration of Gd{sub 3−x}Y{sub x}Co{sub 29}T{sub 4}B{sub 10} (T=Al and Ge)

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

    Cheng, Hai-Xia; Wang, Xiao-Xu; Department of Materials Engineering, National Ping Tung University of Technology and Science, Ping-Tung 91201, Taiwan

    The effects of the Y substitution for Gd on the structural stability and the site preference of intermetallics Gd{sub 3−x}Y{sub x}Co{sub 29}T{sub 4}B{sub 10} (T=Al and Ge) are studied by using a series of interatomic pair potentials. The calculated results show Y can stabilize Gd{sub 3−x}Y{sub x}Co{sub 29}T{sub 4}B{sub 10} with the tetragonal structure, and Y substitute for Gd with a strong preference for the 2b sites. The calculated lattice parameters are in good agreement with the experimental data. Furthermore, the total and partial phonon densities of states are evaluated for the Gd{sub 3−x}Y{sub x}Co{sub 29}T{sub 4}B{sub 10} compounds withmore » the tetragonal structure. A qualitative analysis is carried out with the relevant potentials for the vibrational modes, which makes it possible to predict some properties related to lattice vibration. - Graphical abstract: The lattice cell of Gd{sub 3}Co{sub 29}T{sub 4}B{sub 10} consists of 92 atoms, or two Gd{sub 3}Co{sub 29}T{sub 4}B{sub 10} formula units, with fourteen distinct kinds of site. Rare-earth atoms occupy 2b and 4d sites, Co atoms occupy the Co1(2c), Co2(8i1), Co3(8i2), Co4(8i3), Co5(8j1), Co6(8j2) and Co7(16k), T atoms occupy the T(8i) sites, and B atoms occupy the B1(2c1), B2(2c2), B3(8i) and B4(8j) sites. - Highlights: • The application of the pair potentials obtained from lattice-inversion method. • The lattice vibrations for Gd{sub 3−x}Y{sub x}Co{sub 29}T{sub 4}B{sub 10} (T=Ge and Al) are first evaluated. • The Y atoms should prefer the 2b site of Gd{sub 3−x}Y{sub x}Co{sub 29}T{sub 4}B{sub 10} compounds. • The total and partial phonon densities of states are evaluated for the Gd{sub 3−x}Y{sub x}Co{sub 29}T{sub 4}B{sub 10} compounds with the tetragonal structure. • A qualitative analysis is carried out with the relevant potentials for the vibrational modes.« less

  9. Manipulation of polar order in the “empty” tetragonal tungsten bronzes: Ba{sub 4-x}Sr{sub x}Dy{sub 0.67}□{sub 1.33}Nb{sub 10}O{sub 30}, x = 0, 0.25, 0.5, 1, 2, 3

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

    Gardner, Jonathan; Morrison, Finlay D., E-mail: finlay.morrison@st-andrews.ac.uk

    2016-08-15

    A series of “empty” tetragonal tungsten bronze (TTB) ferroelectrics, Ba{sub 4-x}Sr{sub x}Dy{sub 0.67}□{sub 1.33}Nb{sub 10}O{sub 30} (x = 0, 0.25, 0.5, 1, 2, 3; □ = vacancy), is reported. With increasing x the unit cell contracts in both the ab plane and c-axis; x ≤ 1 compounds are normal ferroelectrics (FE) with decreasing T{sub C} as x increases, while x ≥ 2 are relaxor ferroelectrics (RFE) with associated frequency dependent permittivity peaks and with similar T{sub m} and T{sub f} (Vogel-Fulcher freezing temperatures) values. This observation is rationalised by differing cation occupancies: for x ≤ 1, Sr{sup 2+} principally occupies the A2-site (co-occupied by Ba{sup 2+} with the A1-site occupiedmore » by Dy{sup 3+} and vacancies); for x ≥ 2 significant Sr A1-site occupation leads to the observed RFE characteristics. This FE to RFE crossover is consistent with a previously proposed TTB crystal chemical framework where both a decrease in average A-site size and concurrent increase in A1-site tolerance factor (t{sub A1}) favour destabilization of long range polar order and relaxor behaviour. The effect of increasing t{sub A1} as a result of Sr occupancy at the A1 site is dominant in the compounds reported here.« less

  10. (Pt{sub 1–x}Cu{sub x}){sub 3}Cu{sub 2}B and Pt{sub 9}Cu{sub 3}B{sub 5}, the first examples of copper platinum borides. Observation of superconductivity in a novel boron filled β-Mn-type compound

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

    Salamakha, Leonid P.; Sologub, Oksana, E-mail: oksana.sologub@univie.ac.at; Stöger, Berthold

    New ternary copper platinum borides have been synthesized by arc melting of pure elements followed by annealing at 600 °C. The structures have been studied by X-ray single crystal and powder diffraction. (Pt{sub 1−x}Cu{sub x}){sub 3}Cu{sub 2}B (x=0.33) forms a B-filled β-Mn-type structure (space group P4{sub 1}32; a=0.6671(1) nm). Cu atoms are distributed preferentially on the 8c atom sites, whereas the 12d site is randomly occupied by Pt and Cu atoms (0.670(4) Pt±0.330(4) Cu). Boron is located in octahedral voids of the parent β-Mn-type structure. Pt{sub 9}Cu{sub 3}B{sub 5} (space group P-62m; a=0.9048(3) nm, c=0.2908(1) nm) adopts the Pt{sub 9}Zn{submore » 3}B{sub 5–δ}-type structure. It has a columnar architecture along the short translation vector exhibiting three kinds of [Pt{sub 6}] trigonal prism columns (boron filled, boron semi-filled and empty) and Pt channels with a pentagonal cross section filled with Cu atoms. The striking structural feature is a [Pt{sub 6}] cluster in form of an empty trigonal prism at the origin of the unit cell, which is surrounded by coupled [BPt{sub 6}] and [Pt{sub 6}] trigonal prisms, rotated perpendicularly to the central one. There is no B–B contact as well as Cu–B contact in the structure. The relationships of Pt{sub 9}Cu{sub 3}B{sub 5} structure with the structure of Ti{sub 1+x}Os{sub 2−x}RuB{sub 2} as well as with the structure families of metal sulfides and aluminides have been elucidated. (Pt{sub 1–x}Cu{sub x}){sub 3}Cu{sub 2}B (x=0.3) (B-filled β-Mn-type structure) is a bulk superconductor with a transition temperature of about 2.06 K and an upper critical field μ{sub 0}H{sub C2}(0){sup WHH} of 1.2 T, whereas no superconducting transition has been observed up to 0.3 K in Pt{sub 9}Cu{sub 3}B{sub 5} (Pt{sub 9}Zn{sub 3}B{sub 5–δ}-type structure) from electrical resistivity measurements. - Highlights: • First two copper platinum borides, (Pt{sub 0.67}Cu{sub 0.33}){sub 3}Cu{sub 2}B and Pt{sub 9}Cu{sub

  11. M{sub 2}X intermetallics: Nonmetal insertion in a h.c.-like metallic distribution

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

    Pivan, J.Y.; Guerin, R.

    A simple structural model is proposed on the basis of an ideal hexagonal close-packing (AB){sup {infinity}} of the metal atoms M. The metalloid atoms X, located in an ordered manner in the metallic planes with the stacking sequence ..(A){sup {infinity}}.. or ..(B){sup {infinity}}.., generate two types of elementary units called units U{sub I} and U{sub II}. These units are hexagonal prisms with the fundamental vectors a{sub 0}, b{sub 0}, c{sub 0} and the elementary volume V{sub 0}. When the exclusive occurrence of U{sub I} (or U{sub II}) only yields 6-prismatic and triangular sites of metalloid atoms X, additional tetrahedral andmore » pyramidal sites of X atoms are present when units U{sub I} and U{sub II} exist simultaneously. The structures of compounds with the general formula M{sub 2}X (M = 3d, 4d, 5d and (or) 4f, 5f elements, X = B, P, As, Sb, Si, Ge, {hor_ellipsis}) are described in terms of intergrowth mechanisms. Binary, ternary, or even quaternary structures of compounds, with M/X ratios equal or close to two, appear as superstructures of the elementary units. Insofar as no ordering is considered along the stacking direction, the vectorial combinations of the fundamental vectors in the form A = h.a{sub 0}+s.b{sub 0}, B = k.a{sub 0}+t.b{sub 0}, C = c{sub 0} result in supercells with the volume V = (h.t {minus} k.s).V{sub 0}. The attainable symmetry (hexagonal, tetragonal, orthorhombic, monoclinic, {hor_ellipsis}) strongly depends on the particular values of the integers h, s, k, t. The criteria of occurrence of various series of compounds are presented together with their crystallographic features and structural relationships are emphasized. Moreover, the model demonstrates that numerous compounds with the predicted unit cell parameters should be obtained in each crystal system.« less

  12. Expression, purification and crystallization of two major envelope proteins from white spot syndrome virus

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

    Tang, Xuhua; Hew, Choy Leong, E-mail: dbshewcl@nus.edu.sg

    2007-07-01

    The crystallization of the N-terminal transmembrane region-truncated VP26 and VP28 of white spot syndrome virus is described. White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapour-diffusion method. Crystals ofmore » SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 M sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 Å resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 Å. SeMet-labelled VP28 crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 Å, and diffracts to 2.0 Å resolution.« less

  13. Doxycycline-Regulated 3T3-L1 Preadipocyte Cell Line with Inducible, Stable Expression of Adenoviral E4orf1 Gene: A Cell Model to Study Insulin-Independent Glucose Disposal

    PubMed Central

    Krishnapuram, Rashmi; Dhurandhar, Emily J.; Dubuisson, Olga; Hegde, Vijay; Dhurandhar, Nikhil V.

    2013-01-01

    Impaired glycemic control and excessive adiposity are major risk factors for Type 2 Diabetes mellitus. In rodent models, Ad36, a human adenovirus, improves glycemic control, independent of dietary fat intake or adiposity. It is impractical to use Ad36 for therapeutic action. Instead, we identified that E4orf1 protein of Ad36, mediates its anti-hyperglycemic action independent of insulin signaling. To further evaluate the therapeutic potential of E4orf1 to improve glycemic control, we established a stable 3T3-L1 cell system in which E4orf1 expression can be regulated. The development and characterization of this cell line is described here. Full-length adenoviral-36 E4orf1 cDNA obtained by PCR was cloned into a tetracycline responsive element containing vector (pTRE-Tight-E4orf1). Upon screening dozens of pTRE-Tight-E4orf1 clones, we identified the one with the highest expression of E4orf1 in response to doxycycline treatment. Furthermore, using this inducible system we characterized the ability of E4orf1 to improve glucose disposal in a time dependent manner. This stable cell line offers a valuable resource to carefully study the novel signaling pathways E4orf1 uses to enhance cellular glucose disposal independent of insulin. PMID:23544159

  14. Doxycycline-regulated 3T3-L1 preadipocyte cell line with inducible, stable expression of adenoviral E4orf1 gene: a cell model to study insulin-independent glucose disposal.

    PubMed

    Krishnapuram, Rashmi; Dhurandhar, Emily J; Dubuisson, Olga; Hegde, Vijay; Dhurandhar, Nikhil V

    2013-01-01

    Impaired glycemic control and excessive adiposity are major risk factors for Type 2 Diabetes mellitus. In rodent models, Ad36, a human adenovirus, improves glycemic control, independent of dietary fat intake or adiposity. It is impractical to use Ad36 for therapeutic action. Instead, we identified that E4orf1 protein of Ad36, mediates its anti-hyperglycemic action independent of insulin signaling. To further evaluate the therapeutic potential of E4orf1 to improve glycemic control, we established a stable 3T3-L1 cell system in which E4orf1 expression can be regulated. The development and characterization of this cell line is described here. Full-length adenoviral-36 E4orf1 cDNA obtained by PCR was cloned into a tetracycline responsive element containing vector (pTRE-Tight-E4orf1). Upon screening dozens of pTRE-Tight-E4orf1 clones, we identified the one with the highest expression of E4orf1 in response to doxycycline treatment. Furthermore, using this inducible system we characterized the ability of E4orf1 to improve glucose disposal in a time dependent manner. This stable cell line offers a valuable resource to carefully study the novel signaling pathways E4orf1 uses to enhance cellular glucose disposal independent of insulin.

  15. Protein-protein interface detection using the energy centrality relationship (ECR) characteristic of proteins.

    PubMed

    Sudarshan, Sanjana; Kodathala, Sasi B; Mahadik, Amruta C; Mehta, Isha; Beck, Brian W

    2014-01-01

    Specific protein interactions are responsible for most biological functions. Distinguishing Functionally Linked Interfaces of Proteins (FLIPs), from Functionally uncorrelated Contacts (FunCs), is therefore important to characterizing these interactions. To achieve this goal, we have created a database of protein structures called FLIPdb, containing proteins belonging to various functional sub-categories. Here, we use geometric features coupled with Kortemme and Baker's computational alanine scanning method to calculate the energetic sensitivity of each amino acid at the interface to substitution, identify hotspots, and identify other factors that may contribute towards an interface being FLIP or FunC. Using Principal Component Analysis and K-means clustering on a training set of 160 interfaces, we could distinguish FLIPs from FunCs with an accuracy of 76%. When these methods were applied to two test sets of 18 and 170 interfaces, we achieved similar accuracies of 78% and 80%. We have identified that FLIP interfaces have a stronger central organizing tendency than FunCs, due, we suggest, to greater specificity. We also observe that certain functional sub-categories, such as enzymes, antibody-heavy-light, antibody-antigen, and enzyme-inhibitors form distinct sub-clusters. The antibody-antigen and enzyme-inhibitors interfaces have patterns of physical characteristics similar to those of FunCs, which is in agreement with the fact that the selection pressures of these interfaces is differently evolutionarily driven. As such, our ECR model also successfully describes the impact of evolution and natural selection on protein-protein interfaces. Finally, we indicate how our ECR method may be of use in reducing the false positive rate of docking calculations.

  16. Protein-Protein Interface Detection Using the Energy Centrality Relationship (ECR) Characteristic of Proteins

    PubMed Central

    Sudarshan, Sanjana; Kodathala, Sasi B.; Mahadik, Amruta C.; Mehta, Isha; Beck, Brian W.

    2014-01-01

    Specific protein interactions are responsible for most biological functions. Distinguishing Functionally Linked Interfaces of Proteins (FLIPs), from Functionally uncorrelated Contacts (FunCs), is therefore important to characterizing these interactions. To achieve this goal, we have created a database of protein structures called FLIPdb, containing proteins belonging to various functional sub-categories. Here, we use geometric features coupled with Kortemme and Baker's computational alanine scanning method to calculate the energetic sensitivity of each amino acid at the interface to substitution, identify hotspots, and identify other factors that may contribute towards an interface being FLIP or FunC. Using Principal Component Analysis and K-means clustering on a training set of 160 interfaces, we could distinguish FLIPs from FunCs with an accuracy of 76%. When these methods were applied to two test sets of 18 and 170 interfaces, we achieved similar accuracies of 78% and 80%. We have identified that FLIP interfaces have a stronger central organizing tendency than FunCs, due, we suggest, to greater specificity. We also observe that certain functional sub-categories, such as enzymes, antibody-heavy-light, antibody-antigen, and enzyme-inhibitors form distinct sub-clusters. The antibody-antigen and enzyme-inhibitors interfaces have patterns of physical characteristics similar to those of FunCs, which is in agreement with the fact that the selection pressures of these interfaces is differently evolutionarily driven. As such, our ECR model also successfully describes the impact of evolution and natural selection on protein-protein interfaces. Finally, we indicate how our ECR method may be of use in reducing the false positive rate of docking calculations. PMID:24830938

  17. Recombinant low-seroprevalent adenoviral vectors Ad26 and Ad35 expressing the respiratory syncytial virus (RSV) fusion protein induce protective immunity against RSV infection in cotton rats.

    PubMed

    Widjojoatmodjo, Myra N; Bogaert, Lies; Meek, Bob; Zahn, Roland; Vellinga, Jort; Custers, Jerome; Serroyen, Jan; Radošević, Katarina; Schuitemaker, Hanneke

    2015-10-05

    RSV is an important cause of lower respiratory tract infections in children, the elderly and in those with underlying medical conditions. Although the high disease burden indicates an urgent need for a vaccine against RSV, no licensed RSV vaccine is currently available. We developed an RSV vaccine candidate based on the low-seroprevalent human adenovirus serotypes 26 and 35 (Ad26 and Ad35) encoding the RSV fusion (F) gene. Single immunization of mice with either one of these vectors induced high titers of RSV neutralizing antibodies and high levels of F specific interferon-gamma-producing T cells. A Th1-type immune response was indicated by a high IgG2a/IgG1 ratio of RSV-specific antibodies, strong induction of RSV-specific interferon-gamma and tumor necrosis factor-alpha cytokine producing CD8 Tcells, and low RSV-specific CD4 T-cell induction. Both humoral and cellular responses were increased upon a boost with RSV-F expressing heterologous adenovirus vector (Ad35 boost after Ad26 prime or vice versa). Both single immunization and prime-boost immunization of cotton rats induced high and long-lasting RSV neutralizing antibody titers and protective immunity against lung and nasal RSV A2 virus load up to at least 30 weeks after immunization. Cotton rats were also completely protected against challenge with a RSV B strain (B15/97) after heterologous prime-boost immunization. Lungs from vaccinated animals showed minimal damage or inflammatory infiltrates post-challenge, in contrast to animals vaccinated with formalin-inactivated virus. Our results suggest that recombinant human adenoviral Ad26 and Ad35 vectors encoding the RSV F gene have the potential to provide broad and durable protection against RSV in humans, and appear safe to be investigated in infants. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  18. Advances in Geological CO{sub 2} Sequestration and Co-Sequestration with O{sub 2}

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

    Verba, Circe A; O'Connor, William K.; Ideker, J.H.

    2012-10-28

    The injection of CO{sub 2} for Enhanced Oil Recovery (EOR) and sequestration in brine-bearing formations for long term storage has been in practice or under investigation in many locations globally. This study focused on the assessment of cement wellbore seal integrity in CO{sub 2}- and CO{sub 2}-O{sub 2}-saturated brine and supercritical CO{sub 2} environments. Brine chemistries (NaCl, MgCl{sub 2}, CaCl{sub 2}) at various saline concentrations were investigated at a pressure of 28.9 MPa (4200 psi) at both 50{degree}C and 85{degree}C. These parameters were selected to simulate downhole conditions at several potential CO{sub 2} injection sites in the United States. Classmore » H portland cement is not thermodynamically stable under these conditions and the formation of carbonic acid degrades the cement. Dissociation occurs and leaches cations, forming a CaCO{sub 3} buffered zone, amorphous silica, and other secondary minerals. Increased temperature affected the structure of C-S-H and the hydration of the cement leading to higher degradation rates.« less

  19. Genes on a Wire: The Nucleoid-Associated Protein HU Insulates Transcription Units in Escherichia coli

    PubMed Central

    Berger, Michael; Gerganova, Veneta; Berger, Petya; Rapiteanu, Radu; Lisicovas, Viktoras; Dobrindt, Ulrich

    2016-01-01

    The extent to which chromosomal gene position in prokaryotes affects local gene expression remains an open question. Several studies have shown that chromosomal re-positioning of bacterial transcription units does not alter their expression pattern, except for a general decrease in gene expression levels from chromosomal origin to terminus proximal positions, which is believed to result from gene dosage effects. Surprisingly, the question as to whether this chromosomal context independence is a cis encoded property of a bacterial transcription unit, or if position independence is a property conferred by factors acting in trans, has not been addressed so far. For this purpose, we established a genetic test system assessing the chromosomal positioning effects by means of identical promoter-fluorescent reporter gene fusions inserted equidistantly from OriC into both chromosomal replichores of Escherichia coli K-12. Our investigations of the reporter activities in mutant cells lacking the conserved nucleoid associated protein HU uncovered various drastic chromosomal positional effects on gene transcription. In addition we present evidence that these positional effects are caused by transcriptional activity nearby the insertion site of our reporter modules. We therefore suggest that the nucleoid-associated protein HU is functionally insulating transcription units, most likely by constraining transcription induced DNA supercoiling. PMID:27545593

  20. Quantifying the activity of adenoviral E1A CR2 deletion mutants using renilla luciferase bioluminescence and 3'-deoxy-3'-[18F]fluorothymidine positron emission tomography imaging.

    PubMed

    Leyton, Julius; Lockley, Michelle; Aerts, Joeri L; Baird, Sarah K; Aboagye, Eric O; Lemoine, Nicholas R; McNeish, Iain A

    2006-09-15

    The adenoviral E1A CR2 mutant dl922-947 has potent activity in ovarian cancer. We have used Renilla luciferase bioluminescence imaging to monitor viral E1A expression and replication and [18F]fluorothymidine positron emission tomography ([18F]FLT-PET) to quantify the activity of dl922-947 in vivo. We created dlCR2 Ren, with the same E1A CR2 deletion as dl922-947 and the luciferase gene from Renilla reniformis downstream of E1. Light emitted from s.c. and i.p. IGROV1 ovarian carcinoma xenografts was measured following treatment with dlCR2 Ren. Mice bearing s.c. IGROV1 xenografts were injected with 2.96 to 3.7 MBq of [18F]FLT 48 and 168 hours following i.t. injection of dl922-947 or control virus Ad LM-X. The presence of Renilla luciferase in dlCR2 Ren did not reduce in vitro nor in vivo potency compared with dl922-947. Light emission correlated closely with E1A expression in vitro and peaked 48 hours after dlCR2 Ren injection in both s.c. and i.p. IGROV1 xenografts. It diminished by 168 hours in s.c. tumors but persisted for at least 2 weeks in i.p. models. Normalized tumor [18F]FLT uptake at 60 minutes (NUV60), fractional retention, and area under radioactivity curve all decreased marginally 48 hours after dl922-947 treatment and significantly at 168 hours compared with controls. There was a close linear correlation between NUV60 and both tumor proliferation (Ki67 labeling index) and thymidine kinase 1 expression. Renilla luciferase bioluminescence and [18F]FLT-PET imaging are capable of quantifying the activity and effectiveness of E1A CR2-deleted adenoviral mutants in ovarian cancer.

  1. A Statistical Analysis of Laser Ablated Ba(Sub 0.50)Sr(Sub 0.50)TiO(Sub 3)/LaAlO(Sub 3) Films for Microwave Applications

    NASA Technical Reports Server (NTRS)

    Romanofsky, R. R.; Varaljay, N. C.; Alterovitz, S. A.; Miranda, F. A.; Mueller, C. M.; VanKeuls, F. W.; Kim, J.; Harshavardhan, K. S.

    2002-01-01

    The NASA Glenn Research Center is constructing a 616 element scanning phased array antenna using thin film Ba(sub x)Sr(sub 1-x)TiO(sub 3) based phase shifters. A critical milestone is the production of 616 identical phase shifters at 19 GHz with [asymptotically equal to]4 dB insertion loss and at least 337.5 deg phase shift with 3 percent bandwidth. It is well known that there is a direct relationship between dielectric tuning and loss due to the Kramers-Kronig relationship and that film crystallinity and strain, affected by the substrate template, play an important role. Ba(sub 0.50)Sr(sub 0.50)TiO (sub 3) films, nominally 400 nm thick, were deposited on 48 0.25 mm thick, 5 cm diameter LaAlO(sub 3) wafers. Although previous results suggested that Mn-doped films on MgO were intrinsically superior in terms of phase shift per unit loss, for this application phase shift per unit length was more important. The composition was selected as a compromise between tuning and loss for room temperature operation (e.g. crystallinity progressively degrades for Ba concentrations in excess of 30 percent). As a prelude to fabricating the array, it was necessary to process, screen, and inventory a large number of samples. Variable angle ellipsometry was used to characterize refractive index and film thickness across each wafer. Microstructural properties of the thin films were characterized using high resolution X-ray diffractometry. Finally, prototype phase shifters and resonators were patterned on each wafer and RE probed to measure tuning as a function of dc bias voltage as well as peak (0 field) permittivity and unloaded Q. The relationship among film quality and uniformity and performance is analyzed. This work presents the first statistically relevant study of film quality and microwave performance and represents a milestone towards commercialization of thin ferroelectric films for microwave applications.

  2. A Potent, Imaging Adenoviral Vector Driven by the Cancer-selective Mucin-1 Promoter That Targets Breast Cancer Metastasis

    PubMed Central

    Huyn, Steven T.; Burton, Jeremy B.; Sato, Makoto; Carey, Michael; Gambhir, Sanjiv S.; Wu, Lily

    2009-01-01

    Purpose With breast cancer, early detection and proper staging are critical, and will often influence both the treatment regimen and the therapeutic outcome for those affected with this disease. Improvements in these areas will play a profound role in reducing mortality from breast cancer. Experimental Design In this work we developed a breast cancer – targeted serotype 5 adenoviral vector, utilizing the tumor-specific mucin-1 promoter in combination with the two-step transcriptional amplification system, a system used to augment the activity of weak tissue – specific promoters. Results We showed the strong specificity of this tumor-selective adenovirus to express the luciferase optical imaging gene, leading to diagnostic signals that enabled detection of sentinel lymph node metastasis of breast cancer. Furthermore, we were able to target hepatic metastases following systemic administration of this mucin-1 selective virus. Conclusions Collectively, we showed that the amplified mucin-1 promoter – driven vector is able to deliver to and selectively express a desirable transgene in metastatic lesions of breast tumors. This work has strong clinical relevance to current diagnostic staging approaches, and could add to targeted therapeutic strategies to advance the fight against breast cancer. PMID:19366829

  3. From SO{sub 2} to greenhouse gases: trends and events shaping future emissions trading programs in the United States

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

    Joseph Kruger

    2005-06-15

    Cap-and-trade programs have become widely accepted for the control of conventional air pollution in the United States. However, there is still no political consensus to use these programs to address greenhouse gases. Meanwhile, in the wake of the success of the US SO{sub 2} and NOx trading programs, private companies, state governments, and the European Union are developing new trading programs or other initiatives that may set precedents for a future national US greenhouse gas trading scheme. This paper summarizes the literature on the 'lessons learned' from the SO{sub 2} trading program for greenhouse gas trading, including lessons about themore » potential differences in design that may be necessary because of the different sources, science, mitigation options, and economics inherent in greenhouse gases. The paper discusses how the programs and initiatives mentioned above have been shaped by lessons from past trading programs and whether they are making changes to the SO{sub 2} model to address greenhouse gases. It concludes with an assessment of the implications of these initiatives for a future US national greenhouse gas trading program. 91 refs., 2 tabs.« less

  4. Investigating the effect of V{sub 2}O{sub 5} addition on sodium barium borosilicate glasses

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

    Halder, Rumu, E-mail: rumuhalder24feb@gmail.com; Sengupta, Pranesh; Dey, G. K.

    2016-05-23

    V{sub 2}O{sub 5} doped sodium barium borosilicate glasses were characterized by photoluminescence spectroscopy and electron probe microanalyzer (EPMA). The glass remains homogeneous for lower concentration of V{sub 2}O{sub 5} but a phase separation is observed when V{sub 2}O{sub 5} doping is increased beyond 5 mol%. Detailed microanalysis reveals that the phase separated glass consists of a phase containing V, Ba and Si and a separate Si rich phase within the glass matrix. The luminescence study demonstrated that at low concentration the vanadium mainly interacts with the structural units of B/Si while at higher concentrations, V-O-V/ V-O{sup −} Na{sup +}/Ba{sup 2+} linkagesmore » are formed.« less

  5. Magnetic properties of the layered III-VI diluted magnetic semiconductor Ga{sub 1−x}Fe{sub x}Te

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

    Pekarek, T. M.; Edwards, P. S.; Olejniczak, T. L.

    2016-05-15

    Magnetic properties of single crystalline Ga{sub 1−x}Fe{sub x}Te (x = 0.05) have been measured. GaTe and related layered III-VI semiconductors exhibit a rich collection of important properties for THz generation and detection. The magnetization versus field for an x = 0.05 sample deviates from the linear response seen previously in Ga{sub 1−x}Mn{sub x}Se and Ga{sub 1−x}Mn{sub x}S and reaches a maximum of 0.68 emu/g at 2 K in 7 T. The magnetization of Ga{sub 1−x}Fe{sub x}Te saturates rapidly even at room temperature where the magnetization reaches 50% of saturation in a field of only 0.2 T. In 0.1 T atmore » temperatures between 50 and 400 K, the magnetization drops to a roughly constant 0.22 emu/g. In 0 T, the magnetization drops to zero with no hysteresis present. The data is consistent with Van-Vleck paramagnetism combined with a pronounced crystalline anisotropy, which is similar to that observed for Ga{sub 1−x}Fe{sub x}Se. Neither the broad thermal hysteresis observed from 100-300 K in In{sub 1−x}Mn{sub x}Se nor the spin-glass behavior observed around 10.9 K in Ga{sub 1−x}Mn{sub x}S are observed in Ga{sub 1−x}Fe{sub x}Te. Single crystal x-ray diffraction data yield a rhombohedral space group bearing hexagonal axes, namely R3c. The unit cell dimensions were a = 5.01 Å, b = 5.01 Å, and c = 17.02 Å, with α = 90°, β = 90°, and γ = 120° giving a unit cell volume of 369 Å{sup 3}.« less

  6. Preliminary X-ray crystallographic studies of mouse UPR responsive protein P58(IPK) TPR fragment

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

    Tao, Jiahui; Wu, Yunkun; Ron, David

    2008-02-01

    To investigate the mechanism by which P58(IPK) functions to promote protein folding within the ER, a P58(IPK) TPR fragment without the C-terminal J-domain has been crystallized. Endoplasmic reticulum (ER) stress induces the unfolded protein response (UPR), which can promote protein folding and misfolded protein degradation and attenuate protein translation and protein translocation into the ER. P58(IPK) has been proposed to function as a molecular chaperone to maintain protein-folding homeostasis in the ER under normal and stressed conditions. P58(IPK) contains nine TPR motifs and a C-terminal J-domain within its primary sequence. To investigate the mechanism by which P58(IPK) functions to promotemore » protein folding within the ER, a P58(IPK) TPR fragment without the C-terminal J-domain was crystallized. The crystals diffract to 2.5 Å resolution using a synchrotron X-ray source. The crystals belong to space group P2{sub 1}, with unit-cell parameters a = 83.53, b = 92.75, c = 84.32 Å, α = 90.00, β = 119.36, γ = 90.00°. There are two P58(IPK) molecules in the asymmetric unit, which corresponds to a solvent content of approximately 60%. Structure determination by MAD methods is under way.« less

  7. New μ-SnTe{sub 4} and μ-Sn{sub 2}Te{sub 6} ligands to transition metal: Solvothermal syntheses and characterizations of zinc tellurostannates containing polyamine ligands

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

    Lu, Jialin; Wang, Fang; Shen, Yali

    2014-08-15

    Zinc tellurostannates [(Zn(teta)){sub 2}(μ-1κ:2κ-SnTe{sub 4})] (1), [(Zn(teta)){sub 3}(μ{sub 3}-1κ:2κ:3κ-SnTe{sub 4})]I{sub 2} (2), [(Zn(tren)){sub 2}(μ-1κ:2κ-Sn{sub 2}Te{sub 6})] (3), and [Zn(dien){sub 2}]{sub 4}(Sn{sub 2}Te{sub 6}){sub 1.75}(Sn{sub 2}Te{sub 8}){sub 0.25}·dien (4) were prepared by the reactions of Zn, Sn, and Te with iodine ion assistant in teta and dien. The tetrahedral [SnTe{sub 4}]{sup 4−} anion coordinates to two [Zn(teta)]{sup 2+} units as a bidentate μ-1κ:2κ-SnTe{sub 4} ligand to form the neutral complex 1. It coordinates to three [Zn(teta)]{sup 2+} units with a tridentate μ{sub 3}-1κ:2κ:3κ coordination modes, generating a complex cation [(Zn(teta)){sub 3}(μ{sub 3}-1κ:2κ:3κ- SnTe{sub 4})]{sup 2+} in 2. In 3, the [Sn{submore » 2}Te{sub 6}]{sup 4−} anion joins two [Zn(tren)]{sup 2+}cations with the trans terminal Te atoms, forming neutral complex 3. The μ-1κ:2κ-SnTe{sub 4}, μ{sub 3}-1κ:2κ:3κ-SnTe{sub 4}, and μ-1κ:2κ-Sn{sub 2}Te{sub 6} ligands to TM centers in 1–3 have not been observed before. Compound 4 contains a normal [Sn{sub 2}Te{sub 6}]{sup 4−} and an abnormal [(Sn{sub 2}Te{sub 6}){sub 0.75}(Sn{sub 2}Te{sub 8}){sub 0.25}]{sup 4−} anions. Compounds 1–4 exhibit narrow band gaps in the range of 1.47–1.98 eV, and a distinct red-shift of the band gaps is observed from 4 to 1−3. - Graphical abstract: Zinc tellurostannates were prepared with iodine ion assistant in polyamines, and first μ-1κ:2κ-SnTe{sub 4}, μ{sub 3}-1κ:2κ:3κ-SnTe{sub 4}, and μ-1κ:2κ-Sn{sub 2}Te{sub 6} ligands TM centers were obtained. - Highlights: • Zinc-tellurostannates were first prepared with iodine ion assistant. • Novel μ-1κ:2κ-SnTe{sub 4}, μ{sub 3}-1κ:2κ:3κ-SnTe{sub 4}, and μ-1κ:2κ-Sn{sub 2}Te{sub 6} ligands were obtained. • The Zinc tellurostannates exhibit optical bandgaps between 1.47 and 1.98 eV.« less

  8. Acclimation of respiratory O{sub 2} uptake in green tissues of field-grown native species after long-term exposure to elevated atmospheric CO{sub 2}

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

    Azcon-Bieto, J.; Gonzalez-Meler, M.A.; Doherty, W.

    1994-11-01

    C{sub 3} and C{sub 4} plants were grown in open-top chambers in the field at two CO{sub 2} concentrations, normal ambient (ambient) and normal ambient + 340 {mu}L L{sup {minus}1} (elevated). Dark oxygen uptake was measured in leaves and stems using a liquid-phase Clark-type oxygen electrode. High CO{sub 2} treatment decreased dark oxygen uptake in stems of Scirpus olneyi (C{sub 3}) and leaves of Lindera benzoin (C{sub 3}) expressed on either a dry weight or area basis. Respiration of Spartina patens (C{sub 4}) leaves was unaffected by CO{sub 2} treatment. Leaf dry weight per unit area was unchanged by CO{submore » 2}, but respiration per unit of carbon or per unit of nitrogen was decreased in the C{sub 3} species grown at high CO{sub 2}. The component of respiration in stems of S. olneyi and leaves of L. benzoin primarily affected by long-term exposure to the elevated CO{sub 2} treatment was the activity of the cytochrome pathway. Elevated CO{sub 2} had no effect on activity and capacity of the alternative pathway in S. olneyi. The cytochrome c oxidase activity, assayed in a cell-free extract, was strongly decreased by growth at high CO{sub 2} in stems of S. olneyi but it was unaffected in S. patens leaves. The activity of cytochrome c oxidase and complex III extracted from mature leaves of L. benzoin was also decreased after one growing season of plant exposure to elevated CO{sub 2} concentration. These results show that in some C{sub 3} species respiration will be reduced when plants are grown in elevated atmospheric CO{sub 2}. The possible physiological causes and implications of these effects are discussed. 34 refs., 1 fig., 6 tabs.« less

  9. CO{sub 2}-philic oligomers as novel solvents for CO{sub 2} absorption

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

    Miller, Matthew B; Luebke, David R; Enick, Robert M

    2010-01-01

    Desirable properties for an oligomeric CO{sub 2}-capture solvent in an integrated gasification combined cycle (IGCC) plant include high selectivity for CO{sub 2} over H{sub 2} and water, low viscosity, low vapor pressure, low cost, and minimal environmental, health, and safety impacts. The neat solvent viscosity and solubility of CO{sub 2}, measured via bubble-point loci and presented on a pressure−composition diagram (weight basis), and water miscibility in CO{sub 2}-philic solvents have been determined and compared to results obtained with Selexol, a commercial oligomeric CO{sub 2} solvent. The solvents tested include polyethyleneglycol dimethylether (PEGDME), polypropyleneglycol dimethylether (PPGDME), polypropyleneglycol diacetate (PPGDAc), polybutyleneglycol diacetatemore » (PBGDAc), polytetramethyleneetherglycol diacetate (PTMEGDAc), glyceryl triacetate (GTA), polydimethyl siloxane (PDMS), and perfluorpolyether (PFPE) that has a perfluorinated propyleneglycol monomer unit. Overall, PDMS and PPGDME are the best oligomeric solvents tested and exhibit properties that make them very promising alternatives for the selective absorption of CO{sub 2} from a mixed gas stream, especially if the absorption of water is undesirable.« less

  10. The West. Grade Five (Unit V). Resource Unit. Project Social Studies.

    ERIC Educational Resources Information Center

    Minnesota Univ., Minneapolis. Project Social Studies Curriculum Center.

    This resource unit for 5th graders includes three case studies and a sub-unit on the West as a region. Three sequent occupance case studies which are suggestive, rather than prescriptive, comprise the first part of the unit. Teachers may decide to select only one for an in-depth study or may decide to design a case study modeled after this…

  11. Functional reconstitution of prostaglandin E receptor from bovine adrenal medulla with guanine nucleotide binding proteins

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

    Negishi, M.; Ito, S.; Yokohama, H.

    1988-05-15

    Prostaglandin E/sub 2/ (PEG/sub 2/) was found to bind specifically to a 100,000 x g pellet prepared from bovine adrenal medulla. The PGE receptor was associated with a GTP-binding protein (G-protein) and could be covalently cross-linked with this G-protein by dithiobis(succinimidyl propionate) in the 100,000 x g pellet. In order to characterize the G-protein associated with the PGE receptor and reconstitute these proteins in phospholipid vesicles, the authors purified the G-protein to apparent homogeneity from the 100,000 x g pellet. The G-protein served as a substrate of pertussis toxin but differed in its ..cap alpha.. subunit from two known pertussismore » toxin substrate G-proteins (G/sub i/ and G/sub 0/) purified from bovine brain. The molecular weight of the ..cap alpha.. subunit was 40,000, which is between those of G/sub i/ and G/sub 0/. The purified protein was also distinguished immunologically from G/sub i/ and G/sub 0/ and was referred to as G/sub am/. Reconstitution of the PGE receptor with pure C/sub am/, G/sub i/, or G/sub 0/ in phospholipid vesicles resulted in a remarkable restoration of (/sup 3/H)PGE/sub 2/ binding activity in a GTP-dependent manner. The efficiency of these three G-proteins in this capacity was roughly equal. When pertussis toxin- or N-ethylmaleimide-treated G-proteins, instead of the native ones, were reconstituted into vesicles, the restoration of binding activity was no longer observed. These results indicate that the PGE receptor can couple functionally with G/sub am/, G/sub i/, or G/sub 0/ in phospholipid vesicles and suggest that G/sub am/ may be involved in signal transduction of the PGE receptor in bovine adrenal medulla.« less

  12. Structure determination and optical properties of CsSm(PO{sub 3}){sub 4}

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

    Ben Hassen, N.; Ferhi, M., E-mail: ferhi.mounir@gmail.com; Horchani-Naifer, K.

    2015-03-15

    Graphical abstract: Projection of the CsSm(PO{sub 3}){sub 4} structure viewing along the a axis. - Highlights: • Single crystal of a new polyphosphate CsSm(PO{sub 3}){sub 4} has been synthesized. • The obtained compound has been characterized by several techniques. • The crystal structure of CsSm(PO{sub 3}){sub 4} has been resolved. • Spectroscopic properties of Sm{sup 3+} in CsSm(PO{sub 3}){sub 4} have been performed. - Abstract: A new alkali metal-rare earth polyphosphate CsSm(PO{sub 3}){sub 4} has been synthesized by flux method. The obtained compound has been characterized by means of single crystal X-ray diffraction, Fourier transform infrared (FTIR) and Raman scatteringmore » spectroscopies. It crystallizes in the monoclinic space group P2{sub 1}/n with the following unit-cell parameters: a = 10.382(2), b = 8.978(6), c = 11.205(4) Å, β = 106.398(3)° and Z = 4. The structure of CsSm(PO{sub 3}){sub 4} is an infinite three-dimensional framework made up of double spiral (PO{sub 3}){sub n} chains linked with neighboring SmO{sub 8} and CsO{sub 11} polyhedra. Spectroscopic properties of Sm{sup 3+} in this new compound including excitation, emission, and kinetic measurement have been performed. The emission spectrum shows four transitions characteristics of Sm{sup 3+} in the orange–red region by excitation wavelength at 400 nm. The decay time curve of {sup 4}G{sub 5/2} → {sup 6}H{sub 7/2} transition has been also registered and fitted to a single exponential function.« less

  13. Effect of trypsin inhibitor activity in soya bean on growth performance, protein digestibility and incidence of sub-clinical necrotic enteritis in broiler chicken flocks.

    PubMed

    Palliyeguru, M W C D; Rose, S P; Mackenzie, A M

    2011-06-01

    1. The effect of three different levels of dietary trypsin inhibitor activity (achieved by varying the amount of non-toasted full fat soya bean in replacement for toasted full fat soya bean) on the incidence of spontaneously-occurring sub-clinical necrotic enteritis (NE) in broiler chickens was compared. A fourth dietary treatment compared the effect of a diet that used potato protein concentrate as the major protein source. The determined trypsin inhibitor activity increased with the increasing content of non-toasted soya bean: 1·90, 6·21, 8·46 and 3·72 mg/g for the three soya bean diets (0, 100 and 200 g of non-toasted soya bean/kg) and the potato protein diet respectively. 2. Although increasing amounts of the non-toasted full-fat soya bean increased the feed intakes of the birds, there was a marked reduction in protein digestibility, weight gain and feed conversion efficiency. 3. There was a linear increase in sub-clinical NE lesions in the duodenum, jejunum, mid small intestine and ileum with increasing non-toasted soya bean. Caecal Clostridium perfringens counts increased with the increasing dietary content of non-toasted soya bean. Serum α-toxin antibodies were higher in the birds fed the 200 g non-toasted soya bean/kg diet compared with the other diets. 4. The results demonstrated that variation in the amount of non-toasted dietary soya bean not only affects growth performance of broilers but also affects the incidence of sub-clinical necrotic enteritis in the flock. Ensuring the lowest possible trypsin-inhibitor activity in soya bean samples is a valuable tool to improve the health and welfare of birds and in reducing the financial losses from this disease.

  14. Calculation of k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} for several small detectors and for two linear accelerators using Monte Carlo simulations

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

    Francescon, P.; Cora, S.; Satariano, N.

    2011-12-15

    Purpose: The scope of this study was to determine a complete set of correction factors for several detectors in static small photon fields for two linear accelerators (linacs) and for several detectors. Methods: Measurements for Monte Carlo (MC) commissioning were performed for two linacs, Siemens Primus and Elekta Synergy. After having determined the source parameters that best fit the measurements of field specific output factors, profiles, and tissue-phantom ratio, the generalized version of the classical beam quality correction factor for static small fields, k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}},more » were determined for several types of detectors by using the egs{sub c}hamber Monte Carlo user code which can accurately reproduce the geometry and the material composition of the detector. The influence of many parameters (energy and radial FWHM of the electron beam source, field dimensions, type of accelerator) on the value of k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} was evaluated. Moreover, a MC analysis of the parameters that influence the change of k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} as a function of field dimension was performed. A detailed analysis of uncertainties related to the measurements of the field specific output factor and to the Monte Carlo calculation of k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} was done. Results: The simulations demonstrated that the correction factor k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} can be considered independent from the quality beam factor Q in the range 0.68 {+-} 0.01 for all the detectors analyzed. The k{sub Q{sub c{sub l{sub i{sub

  15. Low temperature structural variations of Na{sub 0.5}Bi{sub 0.5}TiO{sub 3}-7%BaTiO{sub 3} single crystal: Evidences from optical ellipsometry and Raman scattering

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

    Huang, T.; Guo, S.; Xu, L. P.

    2015-06-14

    Optical properties and structural variations of Na{sub 0.5}Bi{sub 0.5}TiO{sub 3}-7%BaTiO{sub 3} (NBT-7%BT) single crystal have been studied by temperature-dependent optical ellipsometry and Raman spectroscopy from 4.2 to 300 K. The second derivative of the complex dielectric functions reveals two interband transitions (E{sub cp1} and E{sub cp2}) located at about 3.49 and 4.25 eV, respectively. Depending on the temperature evolution of electronic transitions, structural variations appear near 60, 150, and 240 K, respectively. These anomalies are also well illustrated from the low-frequency phonon modes involving vibrations of Bi. The low-temperature structural variations of NBT-7%BT crystal can be associated with instability of the crystalline latticemore » driven by off-centered Bi ions, followed by the variations of polarizability of the unit cells.« less

  16. Structure and dynamics of a detergent-solubilized membrane protein: measurement of amide hydrogen exchange rates in M13 coat protein by /sub 1/H NMR spectroscopy

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

    O'Neil, J.D.J.; Sykes, B.D.

    The coat protein of bacteriophage M13 is inserted into the inner membrane of Escherichia coli where it exists as an integral membrane protein during the reproductive cycle of the phage. The protein sequence consists of a highly hydrophobic 19-residue central segment flanked by an acidic 20-residue N-terminus and a basic 11-residue C-terminus. The authors have measured backbone amide hydrogen exchange of the protein solubilized in perdeuteriated sodium dodecyl sulfate using /sup 1/H nuclear magnetic resonance (NMR) spectroscopy. Direct proton exchange-out measurements in D/sub 2/O at 24 /sup 0/C were used to follow the exchange of the slowest amides in themore » protein. Multiple exponential fitting of the exchange data showed that these amides exchanged in two kinetic sets with exchange rates that differed by more than 100-fold. Steady-state saturation-transfer techniques were also used to measure exchange. These methods showed that 15-20 amides in the protein are very stable at 55/sup 0/C and that bout 30 amides have exchange rates retarded by at least 10/sup 5/-fold at 24/sup 0/C. Saturation-transfer studies also showed that the pH dependence of exchange in the hydrophilic termini was unusual. Relaxation and solid-state NMR experiments have previously shown that the majority of the protein backbone is rigid on the picosecond to microsecond time scale, except for the extreme ends of the molecule which are mobile. The hydrogen exchange results, which are sensitive to a much longer time scale, suggest a stable core with a progressive increase in amplitude or frequency of motions as the ends of the protein are approached.« less

  17. Nanoscale morphology of Ni{sub 50}Ti{sub 45}Cu{sub 5} nanoglass

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

    Śniadecki, Z., E-mail: sniadecki@ifmpan.poznan.pl; Institute of Nanotechnology, Karlsruhe Institute of Technology, Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen; Wang, D.

    2016-03-15

    Nanoglasses are noncrystalline solids with a granular nano-/microstructure. In contrast to their nanocrystalline analogs, typically constituted of grains and grain boundaries, nanoglasses consist of glassy regions with a structure corresponding to melt-quenched glasses and amorphous interfaces characterized by a reduced density. Their unique properties can be controlled by modifying size and chemical composition of the granular and interfacial regions. Ni{sub 50}Ti{sub 45}Cu{sub 5} amorphous films were obtained by magnetron sputtering and analyzed to determine their nanoscale morphology and the formation mechanisms. The nanoglasses were noted to have a hierarchical nano-columnar structure with the smallest Ni-rich (Ni:Ti ratio of ca. 5:3)more » amorphous columns with diameters of about 8 nm and Ti-rich glassy interfacial regions with a substantially lower density. The results were obtained utilizing X-ray diffraction and different microscopic methods, e.g., atomic force microscopy and transmission electron microscopy. A detailed analysis indicates the complexity of the formation mechanisms of topologically and chemically distinguishable structural units with curvature driven surface diffusion, surface mobility, self-shadowing and internal stresses as the most important parameters. Common and simple synthesis method and the possibility for easy modification of the morphology and, consequently, the physical properties offer an opportunity for intensive studies of this new class of materials, opening the way towards possible applications. - Highlights: • Ni{sub 50}Ti{sub 45}Cu{sub 5} thin film nanoglasses were synthesized by magnetron sputtering. • Ti amorphous interfacial phase with reduced density is observed. • Stabilization of interfaces by specific local thermodynamic conditions.« less

  18. Scabies in residential care homes: Modelling, inference and interventions for well-connected population sub-units

    PubMed Central

    Middleton, Jo; Güttel, Stefan; Cassell, Jackie; Ross, Joshua

    2018-01-01

    In the context of an ageing population, understanding the transmission of infectious diseases such as scabies through well-connected sub-units of the population, such as residential care homes, is particularly important for the design of efficient interventions to mitigate against the effects of those diseases. Here, we present a modelling methodology based on the efficient solution of a large-scale system of linear differential equations that allows statistical calibration of individual-based random models to real data on scabies in residential care homes. In particular, we review and benchmark different numerical methods for the integration of the differential equation system, and then select the most appropriate of these methods to perform inference using Markov chain Monte Carlo. We test the goodness-of-fit of this model using posterior predictive intervals and propagate forward the resulting parameter uncertainty in a Bayesian framework to consider the economic cost of delayed interventions against scabies, quantifying the benefits of prompt action in the event of detection. We also revisit the previous methodology used to assess the safety of treatments in small population sub-units—in this context ivermectin—and demonstrate that even a very slight relaxation of the implicit assumption of homogeneous death rates significantly increases the plausibility of the hypothesis that ivermectin does not cause excess mortality based upon the data of Barkwell and Shields. PMID:29579037

  19. Phase transformation of Ca{sub 4}[Al{sub 6}O{sub 12}]SO{sub 4} and its disordered crystal structure at 1073 K

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

    Kurokawa, Daisuke; R and D Center, Taiheiyo Cement Corporation, Chiba 285-8655; Takeda, Seiya

    The phase transformation of Ca{sub 4}[Al{sub 6}O{sub 12}]SO{sub 4} and the crystal structure of its high-temperature phase were investigated by differential thermal analysis, temperature-dependent Raman spectroscopy and high-temperature X-ray powder diffraction (CuKα{sub 1}). We determined the starting temperature of the orthorhombic-to-cubic transformation during heating (=711 K) and that of the reverse transformation during cooling (=742 K). The thermal hysteresis was negative (=−31 K), suggesting the thermoelasticity of the transformation. The space group of the high temperature phase is I4{sup ¯}3m with the unit-cell dimensions of a=0.92426(2) nm and V=0.78955(2) nm{sup 3} (Z=2) at 1073 K. The initial structural model wasmore » derived by the direct methods and further refined by the Rietveld method. The final structural model showed the orientational disordering of SO{sub 4} tetrahedra. The maximum-entropy method-based pattern fitting method was used to confirm the validity of the split-atom model, in which conventional structure bias caused by assuming intensity partitioning was minimized. At around the transformation temperature during heating, the vibrational spectra, corresponding to the Raman-active SO{sub 4} internal stretching mode, showed the continuous and gradual change in the slope of full width at half maximum versus temperature curve. This strongly suggests that the orthorhombic-to-cubic phase transformation would be principally accompanied by the statistical disordering in orientation of the SO{sub 4} tetrahedra, without distinct dynamical reorientation. - Graphical abstract: (Left) Three-dimensional electron-density distributions of the SO{sub 4} tetrahedron with the split-atom model, and (right) a bird's eye view of electron densities on the plane parallel to (111). - Highlights: • Crystal structure of Ca{sub 4}[Al{sub 6}O{sub 12}]SO{sub 4} at 1073 K is determined by powder XRD. • The atom arrangements are represented by the split

  20. Integrating Structure to Protein-Protein Interaction Networks That Drive Metastasis to Brain and Lung in Breast Cancer

    PubMed Central

    Engin, H. Billur; Guney, Emre; Keskin, Ozlem; Oliva, Baldo; Gursoy, Attila

    2013-01-01

    Blocking specific protein interactions can lead to human diseases. Accordingly, protein interactions and the structural knowledge on interacting surfaces of proteins (interfaces) have an important role in predicting the genotype-phenotype relationship. We have built the phenotype specific sub-networks of protein-protein interactions (PPIs) involving the relevant genes responsible for lung and brain metastasis from primary tumor in breast cancer. First, we selected the PPIs most relevant to metastasis causing genes (seed genes), by using the “guilt-by-association” principle. Then, we modeled structures of the interactions whose complex forms are not available in Protein Databank (PDB). Finally, we mapped mutations to interface structures (real and modeled), in order to spot the interactions that might be manipulated by these mutations. Functional analyses performed on these sub-networks revealed the potential relationship between immune system-infectious diseases and lung metastasis progression, but this connection was not observed significantly in the brain metastasis. Besides, structural analyses showed that some PPI interfaces in both metastasis sub-networks are originating from microbial proteins, which in turn were mostly related with cell adhesion. Cell adhesion is a key mechanism in metastasis, therefore these PPIs may be involved in similar molecular pathways that are shared by infectious disease and metastasis. Finally, by mapping the mutations and amino acid variations on the interface regions of the proteins in the metastasis sub-networks we found evidence for some mutations to be involved in the mechanisms differentiating the type of the metastasis. PMID:24278371

  1. Growth of Ca{sub 2}MnO{sub 4} Ruddlesden-Popper structured thin films using combinatorial substrate epitaxy

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

    Lacotte, M.; David, A.; Pravarthana, D.

    2014-12-28

    The local epitaxial growth of pulsed laser deposited Ca{sub 2}MnO{sub 4} films on polycrystalline spark plasma sintered Sr{sub 2}TiO{sub 4} substrates was investigated to determine phase formation and preferred epitaxial orientation relationships (ORs) for isostructural Ruddlesden-Popper (RP) heteroepitaxy, further developing the high-throughput synthetic approach called Combinatorial Substrate Epitaxy (CSE). Both grazing incidence X-ray diffraction and electron backscatter diffraction patterns of the film and substrate were indexable as single-phase RP-structured compounds. The optimal growth temperature (between 650 °C and 800 °C) was found to be 750 °C using the maximum value of the average image quality of the backscattered diffraction patterns. Films grew inmore » a grain-over-grain pattern such that each Ca{sub 2}MnO{sub 4} grain had a single OR with the Sr{sub 2}TiO{sub 4} grain on which it grew. Three primary ORs described 47 out of 49 grain pairs that covered nearly all of RP orientation space. The first OR, found for 20 of the 49, was the expected RP unit-cell over RP unit-cell OR, expressed as [100][001]{sub film}||[100][001]{sub sub}. The other two ORs were essentially rotated from the first by 90°, with one (observed for 17 of 49 pairs) being rotated about the [100] and the other (observed for 10 of 49 pairs) being rotated about the [110] (and not exactly by 90°). These results indicate that only a small number of ORs are needed to describe isostructural RP heteroepitaxy and further demonstrate the potential of CSE in the design and growth of a wide range of complex functional oxides.« less

  2. Troponin T3 regulates nuclear localization of the calcium channel Ca{sub v}β{sub 1a} subunit in skeletal muscle

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

    Zhang, Tan; Taylor, Jackson; Jiang, Yang

    The voltage-gated calcium channel (Ca{sub v}) β{sub 1a} subunit (Ca{sub v}β{sub 1a}) plays an important role in excitation–contraction coupling (ECC), a process in the myoplasm that leads to muscle-force generation. Recently, we discovered that the Ca{sub v}β{sub 1a} subunit travels to the nucleus of skeletal muscle cells where it helps to regulate gene transcription. To determine how it travels to the nucleus, we performed a yeast two-hybrid screening of the mouse fast skeletal muscle cDNA library and identified an interaction with troponin T3 (TnT3), which we subsequently confirmed by co-immunoprecipitation and co-localization assays in mouse skeletal muscle in vivo andmore » in cultured C2C12 muscle cells. Interacting domains were mapped to the leucine zipper domain in TnT3 COOH-terminus (160–244 aa) and Ca{sub v}β{sub 1a} NH{sub 2}-terminus (1–99 aa), respectively. The double fluorescence assay in C2C12 cells co-expressing TnT3/DsRed and Ca{sub v}β{sub 1a}/YFP shows that TnT3 facilitates Ca{sub v}β{sub 1a} nuclear recruitment, suggesting that the two proteins play a heretofore unknown role during early muscle differentiation in addition to their classical role in ECC regulation. - Highlights: • Previously, we demonstrated that Ca{sub v}β{sub 1a} is a gene transcription regulator. • Here, we show that TnT3 interacts with Ca{sub v}β{sub 1a}. • We mapped TnT3 and Ca{sub v}β{sub 1a} interaction domain. • TnT3 facilitates Ca{sub v}β{sub 1a} nuclear enrichment. • The two proteins play a heretofore unknown role during early muscle differentiation.« less

  3. The instrument control unit of SPICA SAFARI: a macro-unit to host all the digital control functionalities of the spectrometer

    NASA Astrophysics Data System (ADS)

    Di Giorgio, Anna Maria; Biondi, David; Saggin, Bortolino; Shatalina, Irina; Viterbini, Maurizio; Giusi, Giovanni; Liu, Scige J.; Cerulli-Irelli, Paquale; Van Loon, Dennis; Cara, Christophe

    2012-09-01

    We present the preliminary design of the Instrument Control Unit (ICU) of the SpicA FAR infrared Instrument (SAFARI), an imaging Fourier Transform Spectrometer (FTS) designed to give continuous wavelength coverage in both photometric and spectroscopic modes from around 34 to 210 µm. Due to the stringent requirements in terms of mass and volume, the overall SAFARI warm electronics will be composed by only two main units: Detector Control Unit and ICU. ICU is therefore a macro-unit incorporating the four digital sub-units dedicated to the control of the overall instrument functionalities: the Cooler Control Unit, the Mechanism Control Unit, the Digital processing Unit and the Power Supply Unit. Both the mechanical solution adopted to host the four sub-units and the internal electrical architecture are presented as well as the adopted redundancy approach.

  4. Integrated enzyme reactor and high resolving chromatography in "sub-chip" dimensions for sensitive protein mass spectrometry.

    PubMed

    Hustoft, Hanne Kolsrud; Brandtzaeg, Ole Kristian; Rogeberg, Magnus; Misaghian, Dorna; Torsetnes, Silje Bøen; Greibrokk, Tyge; Reubsaet, Léon; Wilson, Steven Ray; Lundanes, Elsa

    2013-12-16

    Reliable, sensitive and automatable analytical methodology is of great value in e.g. cancer diagnostics. In this context, an on-line system for enzymatic cleavage of proteins, subsequent peptide separation by liquid chromatography (LC) with mass spectrometric detection has been developed using "sub-chip" columns (10-20 μm inner diameter, ID). The system could detect attomole amounts of isolated cancer biomarker progastrin-releasing peptide (ProGRP), in a more automatable fashion compared to previous methods. The workflow combines protein digestion using an 20 μm ID immobilized trypsin reactor with a polymeric layer of 2-hydroxyethyl methacrylate-vinyl azlactone (HEMA-VDM), desalting on a polystyrene-divinylbenzene (PS-DVB) monolithic trap column, and subsequent separation of resulting peptides on a 10 μm ID (PS-DVB) porous layer open tubular (PLOT) column. The high resolution of the PLOT columns was maintained in the on-line system, resulting in narrow chromatographic peaks of 3-5 seconds. The trypsin reactors provided repeatable performance and were compatible with long-term storage.

  5. X-ray diffraction analysis of LiCu{sub 2}O{sub 2} crystals with additives of silver atoms

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

    Sirotinkin, V. P., E-mail: irotinkin.vladimir@mail.ru; Bush, A. A.; Kamentsev, K. E.

    2015-09-15

    Silver-containing LiCu{sub 2}O{sub 2} crystals up to 4 × 8 × 8 mm in size were grown by the crystallization of 80(1-x)CuO · 20{sub x}AgNO{sub 3} · 20Li{sub 2}CO{sub 3} (0 ≤ x ≤ 0.5) mixture melt. According to the X-ray spectral and Rietveld X-ray diffraction data, the maximum amount of silver incorporated in the LiCu{sub 2}O{sub 2} structure is about 4 at % relative to the copper content. It was established that silver atoms occupy statistically crystallographic positions of lithium atoms. The incorporation of silver atoms is accompanied by a noticeable increase in parameter c of the LiCu{sub 2}O{submore » 2} rhombic unit cell, a slight increase in parameter a, and a slight decrease in parameter b.« less

  6. Short, unit-memory, Byte-oriented, binary convolutional codes having maximal free distance

    NASA Technical Reports Server (NTRS)

    Lee, L. N.

    1975-01-01

    It is shown that (n sub 0, k sub 0) convolutional codes with unit memory always achieve the largest free distance among all codes of the same rate k sub 0/n sub 0 and same number 2MK sub 0 of encoder states, where M is the encoder memory. A unit-memory code with maximal free distance is given at each place where this free distance exceeds that of the best code with k sub 0 and n sub 0 relatively prime, for all Mk sub 0 less than or equal to 6 and for R = 1/2, 1/3, 1/4, 2/3. It is shown that the unit-memory codes are byte-oriented in such a way as to be attractive for use in concatenated coding systems.

  7. Structure and physical properties of EuTa{sub 2}O{sub 6} tungsten bronze polymorph

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

    Kolodiazhnyi, T., E-mail: kolodiazhnyi.taras@nims.go.jp; Sakurai, H.; Vasylkiv, O.

    A tetragonal tungsten bronze (TTB) polymorph of EuTa{sub 2}O{sub 6} was prepared and analyzed. EuTa{sub 2}O{sub 6} crystallizes in the centrosymmetric Pnam space group (with unit cell: a = 12.3693, b = 12.4254, and c = 7.7228 Å) isomorphous with orthorhombic β-SrTa{sub 2}O{sub 6}. In contrast to early reports, we see no evidence of deviation from paramagnetic Curie-Weiss behavior among the Eu{sup 2+} 4f{sup 7}spins in EuTa{sub 2}O{sub 6} down to 2 K. Dielectric constant shows a broad peak at ca. 50 K with dielectric dispersion resembling diffuse phase transition. The relaxation time, however, follows a simple (non-freezing) thermally activated process with an activation energy of 92 meV and anmore » attempt frequency of f{sub 0} = 5.79 × 10{sup 12 }Hz. A thermal conductivity of EuTa{sub 2}O{sub 6} shows a low-temperature (T ≈ 30 K) “plateau” region reminiscent of a glass-like behaviour in Nb-based TTB compounds. This behaviour can be attributed to the loosely bound Eu{sup 2+} ions occupying large tricapped trigonal prismatic sites in the EuTa{sub 2}O{sub 6} structure.« less

  8. The SubCons webserver: A user friendly web interface for state-of-the-art subcellular localization prediction.

    PubMed

    Salvatore, M; Shu, N; Elofsson, A

    2018-01-01

    SubCons is a recently developed method that predicts the subcellular localization of a protein. It combines predictions from four predictors using a Random Forest classifier. Here, we present the user-friendly web-interface implementation of SubCons. Starting from a protein sequence, the server rapidly predicts the subcellular localizations of an individual protein. In addition, the server accepts the submission of sets of proteins either by uploading the files or programmatically by using command line WSDL API scripts. This makes SubCons ideal for proteome wide analyses allowing the user to scan a whole proteome in few days. From the web page, it is also possible to download precalculated predictions for several eukaryotic organisms. To evaluate the performance of SubCons we present a benchmark of LocTree3 and SubCons using two recent mass-spectrometry based datasets of mouse and drosophila proteins. The server is available at http://subcons.bioinfo.se/. © 2017 The Protein Society.

  9. Standard Free Droplet Digital Polymerase Chain Reaction as a New Tool for the Quality Control of High-Capacity Adenoviral Vectors in Small-Scale Preparations

    PubMed Central

    Boehme, Philip; Stellberger, Thorsten; Solanki, Manish; Zhang, Wenli; Schulz, Eric; Bergmann, Thorsten; Liu, Jing; Doerner, Johannes; Baiker, Armin E.

    2015-01-01

    Abstract High-capacity adenoviral vectors (HCAdVs) are promising tools for gene therapy as well as for genetic engineering. However, one limitation of the HCAdV vector system is the complex, time-consuming, and labor-intensive production process and the following quality control procedure. Since HCAdVs are deleted for all viral coding sequences, a helper virus (HV) is needed in the production process to provide the sequences for all viral proteins in trans. For the purification procedure of HCAdV, cesium chloride density gradient centrifugation is usually performed followed by buffer exchange using dialysis or comparable methods. However, performing these steps is technically difficult, potentially error-prone, and not scalable. Here, we establish a new protocol for small-scale production of HCAdV based on commercially available adenovirus purification systems and a standard method for the quality control of final HCAdV preparations. For titration of final vector preparations, we established a droplet digital polymerase chain reaction (ddPCR) that uses a standard free-end-point PCR in small droplets of defined volume. By using different probes, this method is capable of detecting and quantifying HCAdV and HV in one reaction independent of reference material, rendering this method attractive for accurately comparing viral titers between different laboratories. In summary, we demonstrate that it is possible to produce HCAdV in a small scale of sufficient quality and quantity to perform experiments in cell culture, and we established a reliable protocol for vector titration based on ddPCR. Our method significantly reduces time and required equipment to perform HCAdV production. In the future the ddPCR technology could be advantageous for titration of other viral vectors commonly used in gene therapy. PMID:25640117

  10. Adenoviral vector-mediated GM-CSF gene transfer improves anti-mycobacterial immunity in mice - role of regulatory T cells.

    PubMed

    Singpiel, Alena; Kramer, Julia; Maus, Regina; Stolper, Jennifer; Bittersohl, Lara Friederike; Gauldie, Jack; Kolb, Martin; Welte, Tobias; Sparwasser, Tim; Maus, Ulrich A

    2018-03-01

    Granulocyte macrophage-colony stimulating factor (GM-CSF) is a hematopoietic growth factor involved in differentiation, survival and activation of myeloid and non-myeloid cells with important implications for lung antibacterial immunity. Here we examined the effect of pulmonary adenoviral vector-mediated delivery of GM-CSF (AdGM-CSF) on anti-mycobacterial immunity in M. bovis BCG infected mice. Exposure of M. bovis BCG infected mice to AdGM-CSF either applied on 6h, or 6h and 7days post-infection substantially increased alveolar recruitment of iNOS and IL-12 expressing macrophages, and significantly increased accumulation of IFNγ pos T cells and particularly regulatory T cells (Tregs). This was accompanied by significantly reduced mycobacterial loads in the lungs of mice. Importantly, diphtheria toxin-induced depletion of Tregs did not influence mycobacterial loads, but accentuated immunopathology in AdGM-CSF-exposed mice infected with M. bovis BCG. Together, the data demonstrate that AdGM-CSF therapy improves lung protective immunity against M. bovis BCG infection in mice independent of co-recruited Tregs, which however critically contribute to limit lung immunopathology in BCG-infected mice. These data may be relevant to the development of immunomodulatory strategies to limit immunopathology-based lung injury in tuberculosis in humans. Copyright © 2017 Elsevier GmbH. All rights reserved.

  11. The Amphipathic Helix of Adenovirus Capsid Protein VI Contributes to Penton Release and Postentry Sorting

    PubMed Central

    Martinez, Ruben; Schellenberger, Pascale; Vasishtan, Daven; Aknin, Cindy; Austin, Sisley; Dacheux, Denis; Rayne, Fabienne; Siebert, Alistair; Ruzsics, Zsolt; Gruenewald, Kay

    2014-01-01

    ABSTRACT Nuclear delivery of the adenoviral genome requires that the capsid cross the limiting membrane of the endocytic compartment and traverse the cytosol to reach the nucleus. This endosomal escape is initiated upon internalization and involves a highly coordinated process of partial disassembly of the entering capsid to release the membrane lytic internal capsid protein VI. Using wild-type and protein VI-mutated human adenovirus serotype 5 (HAdV-C5), we show that capsid stability and membrane rupture are major determinants of entry-related sorting of incoming adenovirus virions. Furthermore, by using electron cryomicroscopy, as well as penton- and protein VI-specific antibodies, we show that the amphipathic helix of protein VI contributes to capsid stability by preventing premature disassembly and deployment of pentons and protein VI. Thus, the helix has a dual function in maintaining the metastable state of the capsid by preventing premature disassembly and mediating efficient membrane lysis to evade lysosomal targeting. Based on these findings and structural data from cryo-electron microscopy, we suggest a refined disassembly mechanism upon entry. IMPORTANCE In this study, we show the intricate connection of adenovirus particle stability and the entry-dependent release of the membrane-lytic capsid protein VI required for endosomal escape. We show that the amphipathic helix of the adenovirus internal protein VI is required to stabilize pentons in the particle while coinciding with penton release upon entry and that release of protein VI mediates membrane lysis, thereby preventing lysosomal sorting. We suggest that this dual functionality of protein VI ensures an optimal disassembly process by balancing the metastable state of the mature adenovirus particle. PMID:25473051

  12. SOXAL combined SO{sub x}/NO{sub x} flue gas control demonstration. Quarterly report, January--March 1993

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

    NONE

    AQUATECH Systems a business unit of Allied-Signal Inc., proposes to demonstrate the technical viability and cost effectiveness of the SOXAL process a combined SO{sub x}/NO{sub x} control process on a 3 MW equivalent flue gas slip stream from Niagara Mohawk Power Corporation, Dunkirk Steam Station Boiler No. 4, a coal fired boiler. The SOXAL process combines 90+% sulfur dioxide removal from the flue gas using a sodium based scrubbing solution and regeneration of the spent scrubbing liquor using AQUATECH Systems` proprietary bipolar membrane technology. This regeneration step recovers a stream of sulfur dioxide suitable for subsequent processing to salable sulfurmore » or sulfuric acid. Additionally 90+% control of NO{sub x} gases can be achieved in combination with conventional urea/methanol injection of NO{sub 2} gas into the duct. The SOXAL process is applicable to both utility and industrial scale boilers using either high or low sulfur coal. The SOXAL demonstration Program began September 10, 1991 and is approximately 22 months in duration. During the 6 months of scheduled operations period, expected to begin January 1992, data will be collected from the SOXAL system to define: SO{sub 2} and NO{sub x} control efficiencies; Current efficiency for the regeneration unit; Sulfate oxidation in the absorber; Make-up reagent rates; Product quality including concentrations and compositions; System integration and control philosophy; and Membrane stability and performance with respect to foulants.« less

  13. Synthesis, crystal structure and electronic structure of the binary phase Rh{sub 2}Cd{sub 5}

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

    Koley, Biplab; Chatterjee, S.; Jana, Partha P., E-mail: ppj@chem.iitkgp.ernet.in

    2017-02-15

    A new phase in the Rh-Cd binary system - Rh{sub 2}Cd{sub 5} has been identified and characterized by single crystal X-ray diffraction and Energy dispersive X-ray analysis. The stoichiometric compound Rh{sub 2}Cd{sub 5} crystallizes with a unit cell containing 14 atoms, in the orthorhombic space group Pbam (55). The crystal structure of Rh{sub 2}Cd{sub 5} can be described as a defect form of the In{sub 3}Pd{sub 5} structure with ordered vacancies, formed of two 2D atomic layers with the stacking sequence: ABAB. The A type layers consist of (3.6.3.6)-Kagomé nets of Cd atoms while the B type layers consist ofmore » (3{sup 5}) (3{sup 7})- nets of both Cd and Rh atoms. The stability of this line phase is investigated by first principle electronic structure calculations on the model of ordered Rh{sub 2}Cd{sub 5}. - Graphical abstract: (3.6.3.6)-Kagomé nets of cadmium atoms (top) and (3{sup 5}) (3{sup 7})- nets of both cadmium and rhodium atoms (bottom) in the structure of Rh{sub 2}Cd{sub 5}.« less

  14. Efficacy and site-specificity of adenoviral vector integration mediated by the phage φC31 integrase.

    PubMed

    Robert, Marc-André; Zeng, Yue; Raymond, Benoît; Desfossé, Laurie; Mairey, Emilie; Tremblay, Jacques P; Massie, Bernard; Gilbert, Rénald

    2012-12-01

    Adenoviral vectors deleted of all their viral genes (helper-dependent [HD]) are efficient gene-transfer vehicles. Because transgene expression is rapidly lost in actively dividing cells, we investigated the feasibility of using phage φC31 integrase (φC31-Int) to integrate an HD carrying an attB site and the puromycin resistance gene into human cells (HeLa) and murine myoblasts (C2C12) by co-infection with a second HD-expressing φC31-Int. Because the HD genome is linear, we also investigated whether its circularization, through expression of Cre using a third HD, affects integration. Efficacy and specificity were determined by scoring the number of puromycin-resistant colonies and by sequencing integration sites. Unexpectedly, circularization of HD was unnecessary and it even reduced the integration efficacy. The maximum integration efficacy achieved was 0.5% in HeLa cells and 0.1% in C2C12 myoblasts. Up to 76% of the integration events occurred at pseudo attP sites and previously characterized hotspots were found. A small (two- to three-fold) increase in the number of γ-H2AX positive foci, accompanied by no noticeable change in γ-H2AX expression, indicated the low genotoxicity of φC31-Int. In conclusion, integration of HD mediated by φC31-Int is an attractive alternative to engineer cells, because it permits site-specific integration of large DNA fragments with low genotoxicity.

  15. On the output factor measurements of the CyberKnife iris collimator small fields: Experimental determination of the k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} correction factors for microchamber and diode detectors

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

    Pantelis, E.; Moutsatsos, A.; Zourari, K.

    Purpose: To measure the output factors (OFs) of the small fields formed by the variable aperture collimator system (iris) of a CyberKnife (CK) robotic radiosurgery system, and determine the k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} correction factors for a microchamber and four diode detectors. Methods: OF measurements were performed using a PTW PinPoint 31014 microchamber, four diode detectors (PTW-60017, -60012, -60008, and the SunNuclear EDGE detector), TLD-100 microcubes, alanine dosimeters, EBT films, and polymer gels for the 5 mm, 7.5 mm, 10 mm, 12.5 mm, and 15 mm irismore » collimators at 650 mm, 800 mm, and 1000 mm source to detector distance (SDD). The alanine OF measurements were corrected for volume averaging effects using the 3D dose distributions registered in polymer gel dosimeters. k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} correction factors for the PinPoint microchamber and the diode dosimeters were calculated through comparison against corresponding polymer gel, EBT, alanine, and TLD results. Results: Experimental OF results are presented for the array of dosimetric systems used. The PinPoint microchamber was found to underestimate small field OFs, and a k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} correction factor ranging from 1.127 {+-} 0.022 (for the 5 mm iris collimator) to 1.004 {+-} 0.010 (for the 15 mm iris collimator) was determined at the reference SDD of 800 mm. The PinPoint k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} correction factor was also found to increase with decreasing SDD; k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} values equal to 1.220 {+-} 0.028 and 1

  16. Microfocus diffraction from different regions of a protein crystal: structural variations and unit-cell polymorphism

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

    Thompson, Michael C.; Cascio, Duilio; Yeates, Todd O.

    Real macromolecular crystals can be non-ideal in a myriad of ways. This often creates challenges for structure determination, while also offering opportunities for greater insight into the crystalline state and the dynamic behavior of macromolecules. To evaluate whether different parts of a single crystal of a dynamic protein, EutL, might be informative about crystal and protein polymorphism, a microfocus X-ray synchrotron beam was used to collect a series of 18 separate data sets from non-overlapping regions of the same crystal specimen. A principal component analysis (PCA) approach was employed to compare the structure factors and unit cells across the datamore » sets, and it was found that the 18 data sets separated into two distinct groups, with largeRvalues (in the 40% range) and significant unit-cell variations between the members of the two groups. This categorization mapped the different data-set types to distinct regions of the crystal specimen. Atomic models of EutL were then refined against two different data sets obtained by separately merging data from the two distinct groups. A comparison of the two resulting models revealed minor but discernable differences in certain segments of the protein structure, and regions of higher deviation were found to correlate with regions where larger dynamic motions were predicted to occur by normal-mode molecular-dynamics simulations. The findings emphasize that large spatially dependent variations may be present across individual macromolecular crystals. This information can be uncovered by simultaneous analysis of multiple partial data sets and can be exploited to reveal new insights about protein dynamics, while also improving the accuracy of the structure-factor data ultimately obtained in X-ray diffraction experiments.« less

  17. Structural basis for the enhanced stability of protein model compounds and peptide backbone unit in ammonium ionic liquids.

    PubMed

    Vasantha, T; Attri, Pankaj; Venkatesu, Pannuru; Devi, R S Rama

    2012-10-04

    Protein folding/unfolding is a fascinating study in the presence of cosolvents, which protect/disrupt the native structure of protein, respectively. The structure and stability of proteins and their functional groups may be modulated by the addition of cosolvents. Ionic liquids (ILs) are finding a vast array of applications as novel cosolvents for a wide variety of biochemical processes that include protein folding. Here, the systematic and quantitative apparent transfer free energies (ΔG'(tr)) of protein model compounds from water to ILs through solubility measurements as a function of IL concentration at 25 °C have been exploited to quantify and interpret biomolecular interactions between model compounds of glycine peptides (GPs) with ammonium based ILs. The investigated aqueous systems consist of zwitterionic glycine peptides: glycine (Gly), diglycine (Gly(2)), triglycine (Gly(3)), tetraglycine (Gly(4)), and cyclic glycylglycine (c(GG)) in the presence of six ILs such as diethylammonium acetate (DEAA), diethylammonium hydrogen sulfate (DEAS), triethylammonium acetate (TEAA), triethylammonium hydrogen sulfate (TEAS), triethylammonium dihydrogen phosphate (TEAP), and trimethylammonium acetate (TMAA). We have observed positive values of ΔG'(tr) for GPs from water to ILs, indicating that interactions between ILs and GPs are unfavorable, which leads to stabilization of the structure of model protein compounds. Moreover, our experimental data ΔG'(tr) is used to obtain transfer free energies (Δg'(tr)) of the peptide backbone unit (or glycyl unit) (-CH(2)C═ONH-), which is the most numerous group in globular proteins, from water to IL solutions. To obtain the mechanism events of the ILs' role in enhancing the stability of the model compounds, we have further obtained m-values for GPs from solubility limits. These results explicitly elucidate that all alkyl ammonium ILs act as stabilizers for model compounds through the exclusion of ILs from model compounds of

  18. Protein kinase B/Akt activates c-Jun NH(2)-terminal kinase by increasing NO production in response to shear stress

    NASA Technical Reports Server (NTRS)

    Go, Y. M.; Boo, Y. C.; Park, H.; Maland, M. C.; Patel, R.; Pritchard, K. A. Jr; Fujio, Y.; Walsh, K.; Darley-Usmar, V.; Jo, H.

    2001-01-01

    Laminar shear stress activates c-Jun NH(2)-terminal kinase (JNK) by the mechanisms involving both nitric oxide (NO) and phosphatidylinositide 3-kinase (PI3K). Because protein kinase B (Akt), a downstream effector of PI3K, has been shown to phosphorylate and activate endothelial NO synthase, we hypothesized that Akt regulates shear-dependent activation of JNK by stimulating NO production. Here, we examined the role of Akt in shear-dependent NO production and JNK activation by expressing a dominant negative Akt mutant (Akt(AA)) and a constitutively active mutant (Akt(Myr)) in bovine aortic endothelial cells (BAEC). As expected, pretreatment of BAEC with the PI3K inhibitor (wortmannin) prevented shear-dependent stimulation of Akt and NO production. Transient expression of Akt(AA) in BAEC by using a recombinant adenoviral construct inhibited the shear-dependent stimulation of NO production and JNK activation. However, transient expression of Akt(Myr) by using a recombinant adenoviral construct did not induce JNK activation. This is consistent with our previous finding that NO is required, but not sufficient on its own, to activate JNK in response to shear stress. These results and our previous findings strongly suggest that shear stress triggers activation of PI3K, Akt, and endothelial NO synthase, leading to production of NO, which (along with O(2-), which is also produced by shear) activates Ras-JNK pathway. The regulation of Akt, NO, and JNK by shear stress is likely to play a critical role in its antiatherogenic effects.

  19. The selective phosphorylation of a guanine nucleotide-binding regulatory protein

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

    Carlson, K.E.

    1989-01-01

    Receptor-activated signal transduction pathways regulate the responsiveness of cells to external stimuli. These transduction pathways themselves are subject to regulation, most commonly by phosphorylation. Guanine nucleotide-binding regulatory proteins (G Proteins), as requisite signal transducing elements for many plasma membrane receptors, are considered likely targets for regulation by phosphorylation. Protein kinase C (PKC) has been shown to phosphorylate the {alpha} subunit of G{sub i} and other G proteins in solution. However, the occurrence of the phosphorylation of G{sub 1} within intact cells in response to activation of PKC has not been rigorously demonstrated. In this thesis, the extent to which themore » {alpha} subunits of G{sub i} undergo phosphorylation within human platelets in response to activation of PKC was examined by means of radiolabeling and immunoprecipitation. Incubation of platelets with phorbol-12-myristate-13-acetate (PMA), a potent activator of PKC, promoted the phosphorylation of several proteins within saponin-permeabilized and intact platelets incubated with ({gamma}{sup 32}P)ATP and ({sup 32}P)H{sub 3}PO{sub 4}, respectively. None of the phosphoproteins, however, were precipitated by either of two antisera containing antibodies differing in specificities for epitopes within G{sub i{alpha}}-despite precipitation of a substantial fraction of the subunit itself. In contrast, other antisera, containing antibodies specific for the recently describe G{sub z{alpha}}, or antibodies for both G{sub z{alpha}} and G{sub i{alpha}}, precipitated a 40-kDa phosphoprotein.« less

  20. Sub-Saharan Africa and the United States. United States Department of State Discussion Paper.

    ERIC Educational Resources Information Center

    Cook, Philip R., Jr.

    Intended to provide background for study and discussion, this publication gives updated information on development in sub-Saharan Africa and provides a basis for understanding U.S. policy toward this vital region. The strengths, problems, politics, natural resources, and language and ethnolinguistic groups of this area of the world and the…

  1. Solutal Convection Around Growing Protein Crystal and Diffusional Purification in Space

    NASA Technical Reports Server (NTRS)

    Lee, Chun P.; Chernov, Alexander A.

    2004-01-01

    At least some protein crystals were found to preferentially trap microheterogeneous impurities. The latter are, for example, dimmer molecules of the crystallizing proteines (e.g. ferritin, lysozyme), or the regular molecules on which surfaces small molecules or ions are adsorbed (e.g. acetilated lysozyme) and modi@ molecular charge. Impurities may induce lattice defects and deteriorate structural resolution. Distribution of impurities between mother solution and gorwing crystal is defined by two interrelated distribution coefficients: kappa = rho(sup c, sub 2) and K = (rho(sup c, sub 2)/rho(sup c, sub 1)/rho(sub 2)/rho(sub 1). Here, rho(sub 2), rho(sub 1) and rho(sup c, sub 2) are densities of impurity (2) and regular protein (1) in solution at the growing interface and within the crystal ("c"). For the microheterogeneous impurities studied, K approx. = 2 - 4, so that kappa approx. - 10(exp 2) - 10(exp 3), since K = kappa (rho(sub 1)/rho(sup c, sub 1) and protein solubility ratio rho(sub 1)/rho(sub=p c, sub 2) much less than 1. Therefore, a crystal growing in absence of convection purifies mother solution around itself, grows cleaner and, probably, more perfect. If convection is present, the solution flow permanently brings new impurities to the crystal. This work theoretically addressed two subjects: 1) onset of convection, 2) distribution of impurities.

  2. Implementation of a recovery-oriented model in a sub-acute Intermediate Stay Mental Health Unit (ISMHU).

    PubMed

    Frost, Barry G; Turrell, Megan; Sly, Ketrina A; Lewin, Terry J; Conrad, Agatha M; Johnston, Suzanne; Tirupati, Srinivasan; Petrovic, Kerry; Rajkumar, Sadanand

    2017-01-03

    An ongoing service evaluation project was initiated following the establishment of a new, purpose-built, 20-bed sub-acute Intermediate Stay Mental Health Unit (ISMHU). This paper: provides an overview of the targeted 6-week program, operating within an Integrated Recovery-oriented Model (IRM); characterises the clients admitted during the first 16 months; and documents their recovery needs and any changes. A brief description of the unit's establishment and programs is initially provided. Client needs and priorities were identified collaboratively using the Mental Health Recovery Star (MHRS) and addressed through a range of in-situ, individual and group interventions. Extracted client and service data were analysed using descriptive statistics, paired t-tests examining change from admission to discharge, and selected correlations. The initial 154 clients (165 admissions, average stay = 47.86 days) were predominately male (72.1%), transferred from acute care (75.3%), with schizophrenia or related disorders (74.0%). Readmission rates within 6-months were 16.2% for acute and 3.2% for sub-acute care. Three MHRS subscales were derived, together with stage-of-change categories. Marked improvements in MHRS Symptom management and functioning were identified (z-change = -1.15), followed by Social-connection (z-change = -0.82) and Self-belief (z-change = -0.76). This was accompanied by a mean reduction of 2.59 in the number of pre-action MHRS items from admission to discharge (z-change = 0.98). Clinician-rated Health of the Nation Outcome Scales (HoNOS) improvements were smaller (z-change = 0.41), indicative of illness chronicity. Staff valued the elements of client choice, the holistic and team approach, program quality, review processes and opportunities for client change. Addressing high-levels of need in the 6-week timeframe was raised as a concern. This paper demonstrates that a recovery-oriented model can be successfully implemented at the

  3. An Adenovirus DNA Replication Factor, but Not Incoming Genome Complexes, Targets PML Nuclear Bodies.

    PubMed

    Komatsu, Tetsuro; Nagata, Kyosuke; Wodrich, Harald

    2016-02-01

    Promyelocytic leukemia protein nuclear bodies (PML-NBs) are subnuclear domains implicated in cellular antiviral responses. Despite the antiviral activity, several nuclear replicating DNA viruses use the domains as deposition sites for the incoming viral genomes and/or as sites for viral DNA replication, suggesting that PML-NBs are functionally relevant during early viral infection to establish productive replication. Although PML-NBs and their components have also been implicated in the adenoviral life cycle, it remains unclear whether incoming adenoviral genome complexes target PML-NBs. Here we show using immunofluorescence and live-cell imaging analyses that incoming adenovirus genome complexes neither localize at nor recruit components of PML-NBs during early phases of infection. We further show that the viral DNA binding protein (DBP), an early expressed viral gene and essential DNA replication factor, independently targets PML-NBs. We show that DBP oligomerization is required to selectively recruit the PML-NB components Sp100 and USP7. Depletion experiments suggest that the absence of one PML-NB component might not affect the recruitment of other components toward DBP oligomers. Thus, our findings suggest a model in which an adenoviral DNA replication factor, but not incoming viral genome complexes, targets and modulates PML-NBs to support a conducive state for viral DNA replication and argue against a generalized concept that PML-NBs target incoming viral genomes. The immediate fate upon nuclear delivery of genomes of incoming DNA viruses is largely unclear. Early reports suggested that incoming genomes of herpesviruses are targeted and repressed by PML-NBs immediately upon nuclear import. Genome localization and/or viral DNA replication has also been observed at PML-NBs for other DNA viruses. Thus, it was suggested that PML-NBs may immediately sense and target nuclear viral genomes and hence serve as sites for deposition of incoming viral genomes and

  4. Sol-gel synthesis of K{sub 3}InF{sub 6} and structural characterization of K{sub 2}InC{sub 10}O{sub 10}H{sub 6}F{sub 9}, K{sub 3}InC{sub 12}O{sub 14}H{sub 4}F{sub 18} and K{sub 3}InC{sub 12}O{sub 12}F{sub 18}

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

    Labeguerie, Jessica; Gredin, Patrick; Marrot, Jerome

    2005-10-15

    K{sub 3}InF{sub 6} is synthesized by a sol-gel route starting from indium and potassium acetates dissolved in isopropanol in the stoichiometry 1:3, with trifluoroacetic acid as fluorinating agent. The crystal structures of the organic precursors were solved by X-ray diffraction methods on single crystals. Three organic compounds were isolated and identified: K{sub 2}InC{sub 10}O{sub 10}H{sub 6}F{sub 9}, K{sub 3}InC{sub 12}O{sub 14}H{sub 4}F{sub 18} and K{sub 3}InC{sub 12}O{sub 12}F{sub 18}. The first one, deficient in potassium in comparison with the initial stoichiometry, is unstable. In its crystal structure, acetate as well as trifluoroacetate anions are coordinated to the indium atom. Themore » two other precursors are obtained, respectively, by quick and slow evaporation of the solution. They correspond to the final organic compounds, which give K{sub 3}InF{sub 6} by decomposition at high temperature. The crystal structure of K{sub 3}InC{sub 12}O{sub 14}H{sub 4}F{sub 18} is characterized by complex anions [In(CF{sub 3}COO){sub 4}(OH{sub x}){sub 2}]{sup (5-2x)-} and isolated [CF{sub 3}COOH{sub 2-x}]{sup (x-1)-} molecules with x=2 or 1, surrounded by K{sup +} cations. The crystal structure of K{sub 3}InC{sub 12}O{sub 12}F{sub 18} is only constituted by complex anions [In(CF{sub 3}COO){sub 6}]{sup 3-} and K{sup +} cations. For all these compounds, potassium cations ensure only the electroneutrality of the structure. IR spectra of K{sub 2}InC{sub 10}O{sub 10}H{sub 6}F{sub 9} and K{sub 3}InC{sub 12}O{sub 12}F{sub 18} were also performed at room temperature on pulverized crystals.« less

  5. Complexation of Sn{sub 2}Se{sub 6} with lanthanide(III) centers influenced by ethylene polyamines: Solvothermal syntheses, crystal structures, and optical properties of lanthanide selenidostannates

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

    Tang, Chunying; Wang, Fang; Chen, Ruihong

    Lanthanide selenidostannates (H{sub 3}O){sub n}[Ce(tepa)(μ-1κ{sup 2}:2κ{sup 2}-Sn{sub 2}Se{sub 6})]{sub n} (1), [(Yb(tepa)(μ-OH)){sub 2}(μ-1κ:2κ-Sn{sub 2}Se{sub 6})]{sub n}·nH{sub 2}O (2), [Htrien]{sub 2}[(Ln(trien)(tren)){sub 2}(μ-1κ:2κ-Sn{sub 2}Se{sub 6})][Sn{sub 2}Se{sub 6}] (Ln=Ce(3), Nd(4)) and [(Yb(dien){sub 2}){sub 2}(μ-OH){sub 2}]Sn{sub 2}Se{sub 6} (5) were solvothermally prepared in different ethylene polyamines. The Sn{sub 2}Se{sub 6} unit connects [Ce(tepa)]{sup 3+} and [(Yb(tepa)(μ-OH)){sub 2}]{sup 4+} fragments with tetradentate μ-1κ{sup 2}Se{sup 1},Se{sup 2}:2κ{sup 2}Se{sup 5},Se{sup 6} and bidentate μ-1κSe{sup 1}:2κSe{sup 5} bridging coordination modes in tepa, to form polymers 1 and 2, respectively. It joins two [Ln(trien)(tren)]{sup 3+} fragments as a μ-1κSe{sup 1}:2κSe{sup 5} ligand to form binuclear complexes 3 and 4more » in trien. Unlike the Sn{sub 2}Se{sub 6} units in 1–4 that bind with Ln(III) centers as Se-donor ligands, the Sn{sub 2}Se{sub 6} unit in 5 exists as a discrete ion. The syntheses of 1–5 show that the ethylene polyamines play an important role in the complexation of Sn{sub 2}Se{sub 6} ligand with Ln(III) centers. Compounds 1–5 exhibit optical band gaps in the range of 2.09–2.42 eV, which are influenced by the complexation of Sn{sub 2}Se{sub 6} with Ln(III) centers. - Graphical abstract: New lanthanide complexes concerning the Sn{sub 2}Se{sub 6} ligand were solvothermally prepared, and the effect of ethylene polyamines on the complexation of Sn{sub 2}Se{sub 6} with Ln(III) centers are observed. Highlights: • Lanthanide complexes concerning the selenidostannates have been solvothermally prepared in different ethylene polyamines. • A tetradentate μ-1κ{sup 2}Se{sup 1},Se{sup 2}:2κ{sup 2}Se{sup 5},Se{sup 6} and a bidentate μ-1κSe{sup 1}:2κSe{sup 5} bridging coordination modes for the Sn{sub 2}Se{sub 6} ligand is obtained. • The complexation of the Sn{sub 2}Se{sub 6} ligand with Ln

  6. Thermoelectric and transport properties of sintered n-type K{sub 8}Ba{sub 16}Ga{sub 40}Sn{sub 96} with type-II clathrate structure

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

    Koda, Shota; Kishimoto, Kengo, E-mail: kkishi@yamaguchi-u.ac.jp; Asada, Hironori

    This clathrate had a maximum dimensionless figure-of-merit, ZT, of 0.93 at 637 K, which was slightly higher than that of 0.83 for the sintered type-VIII clathrate Ba{sub 8}Ga{sub 16}Sn{sub 30}. We investigated the high-temperature thermoelectric properties, transport properties, electronic structures, and thermal stabilities of the clathrates. The type-II clathrate was found to be superior to the type-VIII clathrate as a thermoelectric material; it had a high thermal stability and melting point, 859 K, high mobility, 141 cm{sup 2}V{sup −1}s{sup −1} at 300 K, because of its low inertial mass, and low high-temperature lattice thermal conductivity, approximately 4 mW cm{sup −1}K{sup −1}, resulting frommore » a larger unit cell and weaker bipolar thermal conduction. We discuss these properties in terms of the electronic structure and the differences between the two types of clathrate.« less

  7. Porous framework of T{sub 2}[Fe(CN){sub 6}].xH{sub 2}O with T=Co, Ni, Cu, Zn, and H{sub 2} storage

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

    Avila, M.; Reguera, L.; Rodriguez-Hernandez, J.

    2008-11-15

    The materials under study were prepared from aqueous solutions of ferrocyanic acid and salts of the involved transition metals and their crystal structure solved and refined from X-ray powder diffraction data. Complementary information from thermogravimetric, infrared and Moessbauer data was also used for the structural study. Three different crystal structures were found: hexagonal (P-3) for Zn with the zinc atom coordinated to three N ends of CN groups plus a water molecule, cubic (Pm-3m) for Ni and Cu, and monoclinic (P2{sub 1}/m) for Co. For Ni and Cu the obtained solids have an open channel framework related to 50% ofmore » vacancies for the building unit, [Fe(CN){sub 6}]. In the as-synthesized material the framework free volume is occupied by coordinated and hydrogen-bonded water molecules. These of hexacyanoferrates (II) have received certain attention as prototype of materials for the hydrogen storage. In the anhydrous phase of Ni and Cu, 50% of the metal (T) coordination sites, located at the cavities surface, will be available to interact with the hydrogen molecule. However, when the crystal waters are removed the porous frameworks collapse as it is suggested by H{sub 2} and CO{sub 2} adsorption data. For Co, a structure of stacked layers was found where the cobalt atoms have both tetrahedral and octahedral coordination. The layers remain together through a network of hydrogen-bonding interactions between coordinated and weakly bonded water molecules. No H{sub 2} adsorption was observed in the anhydrous phase of Co. For Zn, the porous framework remains stable on the water removal but with a system of narrow channels and a small available volume, also inaccessible to H{sub 2}. - Graphical abstract: Structure of stacked layers for CO{sub 2}[Fe(CN){sub 6}].xH{sub 2}O.« less

  8. Closely related dermatophyte species produce different patterns of secreted proteins.

    PubMed

    Giddey, Karin; Favre, Bertrand; Quadroni, Manfredo; Monod, Michel

    2007-02-01

    Dermatophytes are the most common infectious agents responsible for superficial mycosis in humans and animals. Various species in this group of fungi show overlapping characteristics. We investigated the possibility that closely related dermatophyte species with different behaviours secrete distinct proteins when grown in the same culture medium. Protein patterns from culture filtrates of several strains of the same species were very similar. In contrast, secreted protein profiles from various species were different, and so a specific signature could be associated with each of the six analysed species. In particular, protein patterns were useful to distinguish Trichophyton tonsurans from Trichophyton equinum, which cannot be differentiated by ribosomal DNA sequencing. The secreted proteases Sub2, Sub6 and Sub7 of the subtilisin family, as well as Mep3 and Mep4 of the fungalisin family were identified. SUB6, SUB7, MEP3 and MEP4 genes were cloned and sequenced. Although the protein sequence of each protease was highly conserved across species, their level of secretion by the various species was not equivalent. These results suggest that a switch of habitat could be related to a differential expression of genes encoding homologous secreted proteins.

  9. The synthesis, single-crystal structure, optical absorption, and resistivity of Th{sub 2}GeSe{sub 5}

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

    Koscielski, Lukasz A.; Malliakas, Christos D.; Sarjeant, Amy A.

    2013-09-15

    The compound Th{sub 2}GeSe{sub 5} has been synthesized by the reaction of the elements at 1273 K. From a single-crystal study Th{sub 2}GeSe{sub 5} crystallizes in the Ba{sub 5}Si{sub 3} structure type with four formula units in the space group D{sup 8}{sub 4h}−P4/ncc of the tetragonal system in a cell with dimensions a=7.4968(4) Å and c=13.6302(9) Å at 100(2) K. From optical absorption measurements Th{sub 2}GeSe{sub 5} is found to have an optical band gap of 1.92 eV (indirect) or 1.98 eV (direct), consistent with its red color. Th{sub 2}GeSe{sub 5} is a wide gap semiconductor, as indicated by itsmore » electrical resistivity at 298 K of 4.37(2)×10{sup 9} Ω cm measured on a single crystal. - Graphical abstract: The structure of Th{sub 2}GeSe{sub 5}. Display Omitted - Highlights: • The new compound Th{sub 2}GeSe{sub 5} was synthesized from the elements and recrystallized from Sb{sub 2}Se{sub 3}. • Th{sub 2}GeSe{sub 5} crystallizes in the Ba{sub 5}Si{sub 3} structure type. • The band gap of Th{sub 2}GeSe{sub 5} is1.92 eV and its resistivity shows it to be a wide gap semiconductor.« less

  10. Synthesis, crystal structures and optical properties of two congruent-melting isotypic diphosphates: LiM{sub 3}P{sub 2}O{sub 7} (M=Na, K)

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

    Shi Yunjing; Wang Ying; Graduate University of the Chinese Academy of Sciences, Beijing 100039

    2013-01-15

    Two new isotypic diphosphates, LiNa{sub 3}P{sub 2}O{sub 7} (1) and LiK{sub 3}P{sub 2}O{sub 7} (2), have been synthesized by conventional solid-state reaction. The single-crystal X-ray structural analyses have shown that they crystallize in the orthorhombic space group C222{sub 1} (No. 20) with the unit cells: a=5.4966(2) A, b=9.1365(4) A, c=12.2764(5) A for compound 1 and a=6.0373(14) A, b=9.339(2) A, c=13.292(3) A for compound 2. The LiM{sub 3}P{sub 2}O{sub 7} (M=Na, K) consist of two-dimensional [LiP{sub 2}O{sub 7}]{sup 3-} layers, which are composed by LiO{sub 4} tetrahedral and diphosphate groups, the Na or K atoms are filled in the interlayers andmore » balance the charge. Second harmonic generation (SHG) on powder samples have been measured using Kurtz and Perry techniques. Thermal analyses, IR spectroscopy, UV-vis-NIR diffuse reflectance spectra, and band structure calculations are performed on the reported compounds. - Graphical Abstract: LiM{sub 3}P{sub 2}O{sub 7} (M=Na, K) consists of a two-dimensional infinite [LiP{sub 2}O{sub 7}]{sup 3-} layer, which is composed by LiO{sub 4} tetrahedra and diphosphate groups. Highlights: Black-Right-Pointing-Pointer LiNa{sub 3}P{sub 2}O{sub 7} and LiK{sub 3}P{sub 2}O{sub 7} are new compounds in the Li{sub 2}O-M{sub 2}O (M=Na, K)-P{sub 2}O{sub 5} systems. Black-Right-Pointing-Pointer Crystal structures of LiNa{sub 3}P{sub 2}O{sub 7} and LiK{sub 3}P{sub 2}O{sub 7} consist of two-dimensional [LiP{sub 2}O{sub 7}]{sup 3-} layers. Black-Right-Pointing-Pointer LiNa{sub 3}P{sub 2}O{sub 7} and LiK{sub 3}P{sub 2}O{sub 7} are congruent melting compounds.« less

  11. On the detection of functionally coherent groups of protein domains with an extension to protein annotation

    PubMed Central

    McLaughlin, William A; Chen, Ken; Hou, Tingjun; Wang, Wei

    2007-01-01

    Background Protein domains coordinate to perform multifaceted cellular functions, and domain combinations serve as the functional building blocks of the cell. The available methods to identify functional domain combinations are limited in their scope, e.g. to the identification of combinations falling within individual proteins or within specific regions in a translated genome. Further effort is needed to identify groups of domains that span across two or more proteins and are linked by a cooperative function. Such functional domain combinations can be useful for protein annotation. Results Using a new computational method, we have identified 114 groups of domains, referred to as domain assembly units (DASSEM units), in the proteome of budding yeast Saccharomyces cerevisiae. The units participate in many important cellular processes such as transcription regulation, translation initiation, and mRNA splicing. Within the units the domains were found to function in a cooperative manner; and each domain contributed to a different aspect of the unit's overall function. The member domains of DASSEM units were found to be significantly enriched among proteins contained in transcription modules, defined as genes sharing similar expression profiles and presumably similar functions. The observation further confirmed the functional coherence of DASSEM units. The functional linkages of units were found in both functionally characterized and uncharacterized proteins, which enabled the assessment of protein function based on domain composition. Conclusion A new computational method was developed to identify groups of domains that are linked by a common function in the proteome of Saccharomyces cerevisiae. These groups can either lie within individual proteins or span across different proteins. We propose that the functional linkages among the domains within the DASSEM units can be used as a non-homology based tool to annotate uncharacterized proteins. PMID:17937820

  12. Crystal structure and luminescence properties of silver in AgM(PO{sub 3}){sub 3} (M = Mg, Zn, Ba) polyphosphates

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

    Belharouak, I.; Parent, C.; Tanguy, B.

    1999-06-01

    The relationships between the crystal structures and the luminescent properties of the AgM(PO{sub 3}){sub 3} (M = Mg, Zn, Ba) polyphosphates are reported in comparison with those of AgPO{sub 3}. The structure of the magnesium and zinc phosphates is characterized by long polyphosphates chains connected to infinite chains of [AgO{sub 6}] and [MO{sub 6}] polyhedra sharing faces. The basic structural phosphate unit in AgBa(PO{sub 3}){sub 3} is a P{sub 3}O{sub 9} ring. Silver atoms are located in distorted octahedral sites. Two types of luminescent centers have been observed. The UV emission observed in all these materials is typical of isolatedmore » Ag{sup +} ions. The visible emission observed only in the zinc phosphate is probably the result of a silver-zinc association. 16 refs., 8 figs., 3 tabs.« less

  13. Protein sectors: evolutionary units of three-dimensional structure

    PubMed Central

    Halabi, Najeeb; Rivoire, Olivier; Leibler, Stanislas; Ranganathan, Rama

    2011-01-01

    Proteins display a hierarchy of structural features at primary, secondary, tertiary, and higher-order levels, an organization that guides our current understanding of their biological properties and evolutionary origins. Here, we reveal a structural organization distinct from this traditional hierarchy by statistical analysis of correlated evolution between amino acids. Applied to the S1A serine proteases, the analysis indicates a decomposition of the protein into three quasi-independent groups of correlated amino acids that we term “protein sectors”. Each sector is physically connected in the tertiary structure, has a distinct functional role, and constitutes an independent mode of sequence divergence in the protein family. Functionally relevant sectors are evident in other protein families as well, suggesting that they may be general features of proteins. We propose that sectors represent a structural organization of proteins that reflects their evolutionary histories. PMID:19703402

  14. SOXAL combined SO{sub x}/NO{sub x} flue gas control demonstration. Quarterly report, July--September 1993

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

    NONE

    AQUATECH Systems, a business unit of Allied-Signal Inc., proposes to demonstrate the technical viability and cost effectiveness of the SOXAL process a combined SO{sub x}/NO{sub x} control process on a 3 MW equivalent flue gas slip stream from Niagara Mohawk Power Corporation Dunkirk Steam Station Boiler No. 4, a coal fired boiler. The SOXAL process combines 90+% sulfur dioxide removal from the flue gas using a sodium based scrubbing solution and regeneration of the spent scrubbing liquor using AQUATECH Systems` proprietary bipolar membrane technology. This regeneration step recovers a stream of sulfur dioxide suitable for subsequent processing to salable sulfurmore » or sulfuric acid. Additionally 90+% control of NO{sub x} gases can be achieved in combination with conventional urea/methanol injection of NO{sub 2} gas into the duct. The SOXAL process is applicable to both utility and industrial scale boilers using either high or lower sulfur coal. The SOXAL demonstration Program began September 10, 1991 and is approximately 26 months in duration. During the 6 months of scheduled operations, between January and July of 1993, data was collected from the SOXAL system to define: SO{sub 2} and NO{sub x} control efficiencies; Current efficiency for the regeneration unit; Sulfate oxidation in the absorber; Make-up reagent rates; Product quality including concentrations and compositions; System integration and control philosophy; and Membrane stability and performance with respect to foulants. The program is expected to be concluded in November 1993.« less

  15. Heterometallic niobium complexes containing the {open_quotes}CpNb{close_quotes} unit. X-ray crystal structures of CpNbCl{sub 2}({mu}-SEt){sub 2}NiCp and CpNb(PhCCPh)({mu}-SPr{sup 1}){sub 2}Mo(CO){sub 4}

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

    Woodward, S.; Curtis, M.D.; Rheingold, A.L.

    1992-06-01

    Cp{sub 2}Nb{sub 2}Cl{sub 2}({mu}-Cl{sub 2})(CO){sub 4} reacts with Cp{sub 2}Ni{sub 2}({mu}-SEt){sub 2} to yield paramagnetic CpNbCl{sub 2}({mu}-SEt){sub 2}NiCp: (g) = 2.004, A = 129 G; space group = P2{sub 1}/c,a = 7.466 (1) {Angstrom}, b = 28.427 (5) {Angstrom}, c = 8.620 (2) {Angstrom}, {beta} = 104.81 (1){degree}. CpNb(PhCCPh)({mu}-SPr{sup i}){sub 2}Mo(CO){sub 4} is synthesized by the addition of LiSPr{sup i} to CpNbCl{sub 2}PhCCPh resulting in formation of CpNb(SPr{sup i}){sub 2}(PhCCPh) followed by the reaction of the thiolate complex with Mo(MeCN){sub 3}(CO){sub 3} and CO (1 equiv). The crystal structure for CpNb(PhCCPh)({mu}-SPr{sup i}){sub 2}Mo(CO){sub 4} is reported and EHMO calculations ofmore » the bonding in CpNbCl{sub 2}({mu}-SEt{sub 2})NiCp were discussed. 27 refs., 5 figs., 5 tabs.« less

  16. Crystallization and preliminary X-ray diffraction studies of NP24-I, an isoform of a thaumatin-like protein from ripe tomato fruits

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

    Ghosh, Raka; Chakrabarti, Chandana, E-mail: chandana.chakrabarti@saha.ac.in

    2005-08-01

    A thaumatin-like antifungal protein, NP24-I, has been isolated from ripe tomato fruits. It was crystallized by the vapour-diffusion method and data were collected to 2.45 Å. The structure was solved by molecular replacement. NP24 is a 24 kDa (207-amino-acid) antifungal thaumatin-like protein (TLP) found in tomato fruits. An isoform of the protein, NP24-I, is reported to play a possible role in ripening of the fruit in addition to its antifungal properties. The protein has been isolated and purified and crystallized by the hanging-drop vapour-diffusion method. The crystals belong to the tetragonal space group P4{sub 3}, with unit-cell parameters a =more » b = 61.01, c = 62.90 Å and one molecule per asymmetric unit. X-ray diffraction data were processed to a resolution of 2.45 Å and the structure was solved by molecular replacement.« less

  17. Sub-nanosecond Yb:KLu(WO<sub>4sub>)>2sub> microchip laser.

    PubMed

    Loiko, P; Serres, J M; Mateos, X; Yumashev, K; Yasukevich, A; Petrov, V; Griebner, U; Aguiló, M; Díaz, F

    2016-06-01

    A diode-pumped Yb:KLu(WO<sub>4sub>)>2sub> microchip laser passively Q-switched by a Cr4+:YAG saturable absorber generated a maximum average output power of 590 mW at 1031 nm with a slope efficiency of 55%. The pulse characteristics were 690 ps/47.6 μJ at a pulse repetition frequency of 12.4 kHz. The output beam had an excellent circular profile with M2<1.05. Yb:KLu(WO<sub>4sub>)>2sub> is very promising for ultrathin sub-ns microchip lasers.

  18. Hydrothermal synthesis and structural analysis of new mixed oxyanion borates: Ba{sub 11}B{sub 26}O{sub 44}(PO{sub 4}){sub 2}(OH){sub 6}, Li{sub 9}BaB{sub 15}O{sub 27}(CO{sub 3}) and Ba{sub 3}Si{sub 2}B{sub 6}O{sub 16}

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

    Heyward, Carla, E-mail: cheywar@clemson.edu; McMillen, Colin D., E-mail: cmcmill@clemson.edu; Kolis, Joseph, E-mail: kjoseph@clemson.edu

    2013-07-15

    Several new borate compounds, Ba{sub 11}B{sub 26}O{sub 44}(PO{sub 4}){sub 2}(OH){sub 6} (1), Li{sub 9}BaB{sub 15}O{sub 27}(CO{sub 3}) (2), and Ba{sub 3}Si{sub 2}B{sub 6}O{sub 16} (3) were synthesized containing other hetero-oxyanion building blocks in addition to the borate frameworks. They were all prepared under hydrothermal conditions and characterized by single crystal and powder X-ray diffraction, and IR spectroscopy. Crystal data: For 1; space group P2{sub 1}/c, a=6.8909 (14) Å, b=13.629 (3) Å, c=25.851 (5) Å, β=90.04 (3)°; For 2; space group P-31c, a=8.8599 (13) Å, c=15.148 (3) Å; For 3; space group P-1, a=5.0414 (10) Å, b=7.5602 (15) Å, c=8.5374 (17)more » Å, α=77.15 (3)°, β=77.84 (3)°, γ=87.41 (3)° for 3. Compounds 1 and 2 contain isolated oxyanions [PO{sub 4}]{sup 3−} and [CO{sub 3}]{sup 2−} respectively, sitting in channels created by the borate framework, while structure 3 has the [SiO{sub 4}]{sup 4−} groups directly bonded to the borate groups creating a B–O–Si framework. - Highlights: • Hydrothermal syntheses of three new mixed oxyanion borates are presented. • Ba{sub 11}B{sub 26}O{sub 44}(PO{sub 4}){sub 2}(OH){sub 6} contains isolated [PO{sub 4}]{sup 3−} in voids of the borate framework. • Li{sub 9}BaB{sub 15}O{sub 27}(CO{sub 3}) contains isolated [CO{sub 3}]{sup 2−} in channels created by the framework. • Ba{sub 3}Si{sub 2}B{sub 6}O{sub 16} has direct bonding of [SiO{sub 4}]{sup 4−} and borates creating a B–O–Si framework.« less

  19. X-ray and neutron diffraction study of the defect crystal structure of the as-grown nonstoichiometric phase Y{sub 0.715}Ca{sub 0.285}F{sub 2.715}

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

    Bolotina, N. B., E-mail: bolotina@ns.crys.ras.ru; Kalyukanov, A. I.; Chernaya, T. S.

    This work begins a series of papers aimed at studying the defect structure of nonstoichiometric phases R{sub 1-y}Ca{sub y}F{sub 3-y} with a tysonite-type (LaF{sub 3}) structure. In the single-crystal structure of Y{sub 0.715}Ca{sub 0.285}F{sub 2.715} with a tysonite-type small unit cell (sp. gr. P6{sub 3}/mmc, a = 3.9095(2) A, c = 6.9829(2) A; Z = 2; R{sub w} = 2.16%), the displacements of Y{sup 3+} cations and F{sup 2-} anions from 6{sub 3} symmetry axes were observed for the first time. The X-ray diffraction pattern shows weak satellites insufficient for structural calculations. The LaF{sub 3} structure type is stabilized upmore » and down on the temperature scale due to anion vacancies and the symmetrizing effect of Ca{sup 2+} cations lying on 6{sub 3} symmetry axes. At 120 Degree-Sign C the fluoride-ion conductivity in the nonstoichiometric phase Y{sub 0.715}Ca{sub 0.285}F{sub 2.715} is five orders of magnitude higher than that in the stoichiometric phase {beta}-YF{sub 3}. The transition to a superionic state is caused by a deviation from stoichiometry and is not associated with reconstructive phase transformation.« less

  20. Genome-wide protein-protein interactions and protein function exploration in cyanobacteria

    PubMed Central

    Lv, Qi; Ma, Weimin; Liu, Hui; Li, Jiang; Wang, Huan; Lu, Fang; Zhao, Chen; Shi, Tieliu

    2015-01-01

    Genome-wide network analysis is well implemented to study proteins of unknown function. Here, we effectively explored protein functions and the biological mechanism based on inferred high confident protein-protein interaction (PPI) network in cyanobacteria. We integrated data from seven different sources and predicted 1,997 PPIs, which were evaluated by experiments in molecular mechanism, text mining of literatures in proved direct/indirect evidences, and “interologs” in conservation. Combined the predicted PPIs with known PPIs, we obtained 4,715 no-redundant PPIs (involving 3,231 proteins covering over 90% of genome) to generate the PPI network. Based on the PPI network, terms in Gene ontology (GO) were assigned to function-unknown proteins. Functional modules were identified by dissecting the PPI network into sub-networks and analyzing pathway enrichment, with which we investigated novel function of underlying proteins in protein complexes and pathways. Examples of photosynthesis and DNA repair indicate that the network approach is a powerful tool in protein function analysis. Overall, this systems biology approach provides a new insight into posterior functional analysis of PPIs in cyanobacteria. PMID:26490033

  1. Structural and electronic properties of Sr{sub x}Ba{sub 1-x}SnO{sub 3} from first principles calculations

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

    Moreira, E.; Henriques, J.M.; Azevedo, D.L.

    2012-03-15

    Neutron diffraction data for Sr{sub x}Ba{sub 1-x}SnO{sub 3} (x=0.0, 0.2, 0.4, 0.6, 0.8 and 1.0) solid solutions were used as inputs to obtain optimized geometries and electronic properties using the density functional theory (DFT) formalism considering both the local density and generalized gradient approximations, LDA and GGA, respectively. The crystal structures and SnO{sub 6} octahedra tilting angles found after total energy minimization agree well with experiment, specially for the GGA data. Elastic constants were also obtained and compared with theoretical and experimental results for cubic BaSnO{sub 3}. While the alloys with cubic unit cell have an indirect band gap, tetragonalmore » and orthorhombic alloys exhibit direct band gaps (exception made to x=1.0). The Kohn-Sham minimum electronic band gap oscillates from 1.52 eV (cubic x=0.0, LDA) to 2.61 eV (orthorhombic x=1.0, LDA), and from 0.74 eV (cubic BaSnO{sub 3}, GGA) to 1.97 eV (orthorhombic SrSnO{sub 3}, GGA). Parabolic interpolation of bands has allowed us to estimate the effective masses for charge carriers, which are shown to be anisotropic and larger for holes. - Graphical Abstract: Highlights: Black-Right-Pointing-Pointer DFT calculations were performed on Sr{sub x}Ba{sub 1-x}SnO{sub 3} solid solutions. Black-Right-Pointing-Pointer Calculated crystal structures agree well with experiment. Black-Right-Pointing-Pointer Alloys have direct or indirect gaps depending on the Sr molar fraction. Black-Right-Pointing-Pointer The Kohn-Sham gap variation from x=0.0 to x=1.0 is close to the experimental value. Black-Right-Pointing-Pointer Carrier effective masses are very anisotropic, specially for holes.« less

  2. Erbium-doped borosilicate glasses containing various amounts of P{sub 2}O{sub 5} and Al{sub 2}O{sub 3}: Influence of the silica content on the structure and thermal, physical, optical and luminescence properties

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

    Bourhis, Kevin; Massera, Jonathan; BioMediTech, Tampere

    2015-10-15

    Highlights: • Er{sup 3+} doped borosilicate glasses were processed with different compositions and characterizations. • An increase in the SiO{sub 2} content leads to a silicate-rich environment around the Er{sup 3+} site. • An increase in the SiO{sub 2} content decreases the Er{sup 3+} absorption cross-section at 980 nm. • Glasses with 60 mol% of SiO{sub 2} exhibit a stronger emission intensity at 1530 nm than glasses with x = 50. • Highest 1.5 μm emission intensity was achieved for the Al and P containing glass with 60 mol% of SiO{sub 2}. - Abstract: The influence of the silica contentmore » on several properties of Er-doped borosilicate glasses in the presence of various amounts of P{sub 2}O{sub 5} and Al{sub 2}O{sub 3} has been investigated. The introduction of P{sub 2}O{sub 5} and/or Al{sub 2}O{sub 3} are responsible for structural modifications in the glass network through a charge-compensation mechanism related to the formation of negatively-charged PO{sub 4} and AlO{sub 4} groups or through the formation of AlPO{sub 4}-like structural units. In this paper, we show that an increase in the SiO{sub 2} content leads to a silicate-rich environment around the Er{sup 3+} site, resulting in an increased dependence of the Er{sup 3+} ions optical and luminescence properties on the P{sub 2}O{sub 5} and/or Al{sub 2}O{sub 3} concentration. The highest emission intensity at 1.5 μm was achieved for the glass with an equal proportion of P and Al in the glass system with 60 mol% of SiO{sub 2}.« less

  3. RFe{sub 2}Mg{sub x}Al{sub 8−x} (R=La–Nd and Sm; x≈0.8): Flux synthesis, structure, magnetic and electrical properties

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

    Ma, Xiaowei; Chai, Ping; Chen, Banghao

    2015-09-15

    Single crystals of Mg-substituted CeFe{sub 2}Al{sub 8} type intermetallics RFe{sub 2}Mg{sub x}Al{sub 8–x} (R=La–Nd and Sm; x≤1) were grown by reacting iron and rare earth metals in 1:1 Mg/Al mixed flux. The structure features mono-capped and bi-capped trigonal prismatic FeAl{sub 6} units. Electronic structure calculations indicate that magnesium substitution reduces the valence electron count, shifting the Fermi level away from a pseudo-gap. This changes the electronic nature of the cerium analog; the previously reported ternary CeFe{sub 2}Al{sub 8} shows strong hybridization between the cerium states and the conduction electrons, resulting in no magnetic moment on Ce atoms. On the othermore » hand, magnetic susceptibility measurements on CeFe{sub 2}Mg{sub x}Al{sub 8–x} indicates a localized moment on cerium. The newly synthesized Pr, Nd and Sm analogs exhibit antiferromagnetic ordering at 2.8 K, 7.8 K and 12 K respectively. Solid state {sup 27}Al NMR of LaFe{sub 2}Mg{sub x}Al{sub 8–x} exhibits a broad Knight shift at ~1200 ppm, consistent with the metallic behavior shown by electrical resistivity data. - Graphical abstract: Mg substitution into CeFe{sub 2}Al{sub 8} modifies cerium valence due to changing valence electron count. - Highlights: • RFe{sub 2}Mg{sub x}Al{sub 8−x} (R=La–Nd, Sm) grow as large crystals from reactions in Mg/Al flux. • Products are magnesium-substituted variants of CeFe{sub 2}Al{sub 8}, with CaCo{sub 2}Al{sub 8} structure. • Ce magnetic moment in CeFe{sub 2}Mg{sub x}Al{sub 8−x} varies from that in CeFe{sub 2}Al{sub 8} due to VEC change. • Antiferromagnetic ordering observed for Pr, Nd, Sm analogs of RFe{sub 2}Mg{sub x}Al{sub 8−x}.« less

  4. Transmission electron microscopy study of the MgS–Tm{sub 2}S{sub 3} system

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

    Varadé-López, R., E-mail: rebeca.varade@ucm.es; Ávila-Brande, D., E-mail: davilabr@ucm.es; Urones-Garrote, E., E-mail: esteban.urones@pdi.ucm.es

    2015-09-15

    This work presents the structural–microstructural characterization of the NaCl-derivative MgS–Tm{sub 2}S{sub 3} system, which can be formulated by the expression Mg{sub (1−x)}Tm{sub (2/3)x}□{sub (1/3)x}S (□→cation vacancy). Transmission electron microscopy observations show the transition between NaCl-type and spinel-type structures when 0 ≤x≤ 0.75. The increase of Tm content in the solid solution provokes the increase of the spinel-type phase proportion, which intergrows with the NaCl-type crystals. When x≥0.75, some phases derived from NaCl-type structure through the chemical twinning at the unit cell level crystallographic operation are observed, such as CT-MgTm{sub 2}S{sub 4} and CT-MgTm{sub 4}S{sub 7}. The existence and nature ofmore » the extended defects observed along the c direction of these structures are characterized by means of Scanning-Transmission electron microscopy high-angle dark field imaging, which allows observing the presence of quasi ordered crystals with new possible complex stoichiometries at atomic resolution. - Graphical abstract: HAADF-STEM image of a disordered CT-MgYb{sub 2}S{sub 4} crystal. The disordered twin-slab sequences are marked by arrows. - Highlights: • Structural evolution of the Mg{sub (1−x)}Tm{sub (2/3)x}□{sub (1/3)x}S system was characterized by means of TEM. • The increase in Tm content provokes the transition from NaCl to spinel-type structure up to x=0.75. • Chemical twinned phases CT-MgTm{sub 2}S{sub 4} and CT-MgTm{sub 4}S{sub 7} are observed at high Tm contents. • Extended defects in CT-crystals are characterized with atomic resolution STEM-HAADF images.« less

  5. Pb{sub 5}Fe{sub 3}TiO{sub 11}Cl: A rare example of Ti(IV) in a square pyramidal oxygen coordination

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

    Batuk, Maria, E-mail: Maria.Batuk@uantwerpen.be; Batuk, Dmitry; Abakumov, Artem M.

    A new oxychloride Pb{sub 5}Fe{sub 3}TiO{sub 11}Cl has been synthesized using the solid state method. Its crystal and magnetic structure was investigated in the 1.5–550 K temperature range using electron diffraction, high angle annular dark field scanning transmission electron microscopy, atomic resolution energy dispersive X-ray spectroscopy, neutron and X-ray powder diffraction. At room temperature Pb{sub 5}Fe{sub 3}TiO{sub 11}Cl crystallizes in the P4/mmm space group with the unit cell parameters a=3.91803(3) Å and c=19.3345(2) Å. Pb{sub 5}Fe{sub 3}TiO{sub 11}Cl is a new n=4 member of the oxychloride perovskite-based homologous series A{sub n+1}B{sub n}O{sub 3n−1}Cl. The structure is built of truncated Pb{submore » 3}Fe{sub 3}TiO{sub 11} quadruple perovskite blocks separated by CsCl-type Pb{sub 2}Cl slabs. The perovskite blocks consist of two layers of (Fe,Ti)O{sub 6} octahedra sandwiched between two layers of (Fe,Ti)O{sub 5} square pyramids. The Ti{sup 4+} cations are preferentially located in the octahedral layers, however, the presence of a noticeable amount of Ti{sup 4+} in a five-fold coordination environment has been undoubtedly proven using neutron powder diffraction and atomic resolution compositional mapping. Pb{sub 5}Fe{sub 3}TiO{sub 11}Cl is antiferromagnetically ordered below 450(10) K. The ordered Fe magnetic moments at 1.5 K are 4.06(4) μ{sub B} and 3.86(5) μ{sub B} on the octahedral and square-pyramidal sites, respectively. - Highlights: • Pb{sub 5}Fe{sub 3}TiO{sub 11}Cl has been synthesized using the solid state method. • The structure has been refined using neutron powder diffraction data at 1.5–550 K. • It is a new n=4 member of the perovskite-related homologous series A{sub n+1}B{sub n}O{sub 3n−1}Cl. • Ti{sup 4+} cations have both octahedral and square-pyramidal coordination environment. • Pb{sub 5}Fe{sub 3}TiO{sub 11}Cl is antiferromagnetically ordered below T{sub N}≈450 K.« less

  6. A solid solution series of atacamite type Ni{sub 2x}Mg{sub 2−2x}Cl(OH){sub 3}

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

    Bette, Sebastian; Dinnebier, Robert E.; Röder, Christian

    2015-08-15

    For the first time a complete solid solution series Ni{sub 2x}Mg{sub 2−2x}Cl(OH){sub 3} of an atacamite type alkaline main group metal chloride, Mg{sub 2}Cl(OH){sub 3}, and a transition group metal chloride, Ni{sub 2}Cl(OH){sub 3}, was prepared and characterized by chemical and thermal analysis as well as by Raman and IR spectroscopy, and high resolution laboratory X-ray powder diffraction. All members of the solid solution series crystallize in space group Pnam (62). The main building units of these crystal structures are distorted, edge-linked Ni/MgO{sub 4}Cl{sub 2} and Ni/MgO{sub 5}Cl octahedra. The distribution of Ni{sup 2+}- and Mg{sup 2+}-ions among these twomore » metal-sites within the solid solution series is discussed in detail. The crystallization of the solid solution phases occurs via an intermediate solid solution series, (Ni/Mg)Cl{sub 2x}(OH){sub 2−2x}, with variable Cl: OH ratio up to the 1:3 ratio according to the formula Ni{sub 2x}Mg{sub 2−2x} Cl(OH){sub 3}. For one isolated intermediate solid solution member, Ni{sub 0.70}Mg{sub 0.30}Cl{sub 0.58}(OH){sub 1.42}, the formation and crystal structure is presented as well. - Graphical abstract: For the first time a complete solid solution series, Ni{sub 2x}Mg{sub 2−2x} Cl(OH){sub 3}, was synthesized and characterized. Structure solution by revealed that Ni{sup 2+} prefers to occupy the Jahn–Teller-like distorted hole, out of two available cation sites. Substitution of Ni{sup 2+} by Mg{sup 2+} in atacamite type Ni{sub 2}Cl(OH){sub 3} results in systematic band shifts in Raman and IR spectra as well as in systematic changes in thermal properties. The α-polymorphs M{sub 2}Cl(OH){sub 3} with M=Mg{sup 2+}, Ni{sup 2+} and other divalent transition metal ions, as described in literature, were identified as separate compounds. - Highlights: • First synthesis of solid solution series between main and transition metal chloride. • Ni{sup 2+} prefers to occupy Jahn–Teller-like distorted

  7. Identification, expression, and pharmacology of a Cys{sub 23}-Ser{sub 23} substitution in the human 5-HT{sub 2C} receptor gene (HTR2C)

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

    Lappalainen, J.; Ozaki, N.; Goldman, D.

    1995-05-20

    The function of brain serotonin-2C (5-HT{sub 2C}) receptors, including behavioral and neurochemical responses to 5-HT{sub 2C} agonist challenge, has been suggested to be abnormal in individuals with neuropsychiatric disorders. Thus, it is important to identify polymorphisms and functional variants within this gene. Using SSCP analysis, the authors identified a Cys{sub 23}-Ser{sub 23} substitution (designated 5-HT{sub 2Ccys} and 5-HT{sub 2Cser}) in the first hydrophobic region of the human 5-HT{sub 2C} receptor. Allele frequencies in unrelated Caucasians were 0.13 and 0.87 for 5-HT{sub 2Cser} and 5-HT{sub 2Ccys}, respectively. DNAs from informative CEPH families were typed for this polymorphism and analyzed with respectmore » to 20 linked markers on the X chromosome. Linkage analysis placed the 5-HT{sub 2C} receptor gene (HTR2C) on Xq24. To evaluate whether this amino acid substitution causes a variant function of this receptor, recombinant human 5-HT{sub 2Ccys} and 5-HT{sub 2Cser} receptors were expressed in Xenopus oocytes and tested for responses to 5-HT using electrophysiological techniques. Concentration-response curves for 5-HT were not significantly different in oocytes expressing either form of the receptor, suggesting that the 5-HT{sub 2Ccys} and 5-HT{sub 2Cser} receptor proteins may not differ in their responses to serotonin under baseline physiological conditions. 43 refs., 3 figs., 1 tab.« less

  8. Concerted evolution of the tandemly repeated genes encoding primate U2 small nuclear RNA (the RNU2 locus) does not prevent rapid diversification of the (CT){sub n} {center_dot} (GA){sub n} microsatellite embedded within the U2 repeat unit

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

    Liao, D.; Weiner, A.M.

    1995-12-10

    The RNU2 locus encoding human U2 small nuclear RNA (snRNA) is organized as a nearly perfect tandem array containing 5 to 22 copies of a 5.8-kb repeat unit. Just downstream of the U2 snRNA gene in each 5.8-kb repeat unit lies a large (CT){sub n}{center_dot}(GA){sub n} dinucleotide repeat (n {approx} 70). This form of genomic organization, in which one repeat is embedded within another, provides an unusual opportunity to study the balance of forces maintaining the homogeneity of both kinds of repeats. Using a combination of field inversion gel electrophoresis and polymerase chain reaction, we have been able to studymore » the CT microsatellites within individual U2 tandem arrays. We find that the CT microsatellites within an RNU2 allele exhibit significant length polymorphism, despite the remarkable homogeneity of the surrounding U2 repeat units. Length polymorphism is due primarily to loss or gain of CT dinucleotide repeats, but other types of deletions, insertions, and substitutions are also frequent. Polymorphism is greatly reduced in regions where pure (CT){sub n} tracts are interrupted by occasional G residues, suggesting that irregularities stabilize both the length and the sequence of the dinucleotide repeat. We further show that the RNU2 loci of other catarrhine primates (gorilla, chimpanzee, ogangutan, and baboon) contain orthologous CT microsatellites; these also exhibit length polymorphism, but are highly divergent from each other. Thus, although the CT microsatellite is evolving far more rapidly than the rest of the U2 repeat unit, it has persisted through multiple speciation events spanning >35 Myr. The persistence of the CT microsatellite, despite polymorphism and rapid evolution, suggests that it might play a functional role in concerted evolution of the RNU2 loci, perhaps as an initiation site for recombination and/or gene conversion. 70 refs., 5 figs.« less

  9. Synthesis of cationic iridium(I) complexes of water-soluble phosphine ligands, [Ir(CO)(TPPMS){sub 3}]CF{sub 3}SO{sub 3}, [Ir(CO)(H{sub 2}O)(TPPTS){sub 2}]CF{sub 3}SO{sub 3}, and [Ir(CO){sub 2}(TPPMS){sub 3}]ClO{sub 4} (TPPMS = PPh{sub 2}(m-C{sub 6}H{sub 4}SO{sub 3}K), TPPTS = P(m-C{sub 6}H{sub 4}SO{sub 3}Na){sub 3})

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

    Paterniti, D.P.; Francisco, L.W.; Atwood, J.D.

    Several new water-soluble iridium(I) complexes were synthesized and their reactivities with small molecules (H{sub 2} or CO) in polar solvents (DMSO or H{sub 2}O) examined. Reaction of H{sub 2} with [Ir(CO)(TPPMS){sub 3}]CF{sub 3}SO{sub 3} (TPPMS = P(C{sub 6}H{sub 5}){sub 2}(m-C{sub 6}H{sub 4}SO{sub 3}K)) in DMSO or H{sub 2}O produces [cis,mer-Ir(CO)(H){sub 2}(TPPMS){sub 3}]CF{sub 3}SO{sub 3}, while the reaction of CO with [Ir(CO)(TPPMS){sub 3}]-CF{sub 3}SO{sub 3} in water yields [Ir(CO){sub 2}(TPPMS){sub 3}]CF{sub 3}SO{sub 3}. Carbonylation of [Ir(CO){sub 2}(TPPMS){sub 3}]ClO{sub 4} in DMSO produces [Ir(CO){sub 3}(TPPMS){sub 2}]ClO{sub 4} and TPPMS; no reaction is observed in H{sub 2}O. Hydrogenation of [Ir(CO){sub 2}(TPPMS){sub 3}]ClO{sub 4}more » in DMSO or H{sub 2}O yields [cis,mer-Ir(CO)(H){sub 2}(TPPMS){sub 3}]ClO{sub 4}, while reaction of H{sub 2} with an aqueous solution of [Ir(CO)(H{sub 2}O)(TPPTS){sub 2}]CF{sub 3}SO{sub 3} produces [Ir(CO)(H{sub 2}O)(H){sub 2}(TPPTS){sub 2}]CF{sub 3}SO{sub 3}. Reaction of trans-Ir(CO)ClL{sub 2} (L = TPPMS or TPPTS) with excess L in H{sub 2}O produces [Ir(CO)L{sub 3}]Cl, while no reaction occurs in DMSO, [Ir(CO){sub 3}(TPPMS){sub 2}]Cl reacts irreversibly with TPPMS in H{sub 2}O to produce [Ir(CO){sub 2}-(TPPMS){sub 3}]Cl.« less

  10. Phase equilibria in the quasiternary system Ag{sub 2}S–Ga{sub 2}S{sub 3}–In{sub 2}S{sub 3} and optical properties of (Ga{sub 55}In{sub 45}){sub 2}S{sub 300}, (Ga{sub 54.59}In{sub 44.66}Er{sub 0.75}){sub 2}S{sub 300} single crystals

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

    Ivashchenko, I.A., E-mail: Ivashchenko.Inna@eenu.edu.ua; Danyliuk, I.V.; Olekseyuk, I.D.

    The quasiternary system Ag{sub 2}S–Ga{sub 2}S{sub 3}–In{sub 2}S{sub 3} was investigated by differential thermal, X-ray diffraction analyses. The phase diagram of the Ga{sub 2}S{sub 3}–In{sub 2}S{sub 3} system and nine polythermal sections, isothermal section at 820 K and the liquidus surface projection were constructed. The existence of the large solid solutions ranges of binary and ternary compounds was established. The range of the existence of the quaternary phase AgGa{sub x}In{sub 5−x}S{sub 8} (2.25≤x≤2.85) at 820 K was determined. The single crystals (Ga{sub 55}In{sub 45}){sub 2}S{sub 300} and (Ga{sub 54.59}In{sub 44.66}Er{sub 0.75}){sub 2}S{sub 300} were grown by a directional crystallization methodmore » from solution-melt. Optical absorption spectra in the 500–1600 nm range were recorded. The luminescence of the (Ga{sub 54.59}In{sub 44.66}Er{sub 0.75}){sub 2}S{sub 300} single crystal shows a maximum at 1530 nm for the excitation wavelengths of 532 and 980 nm at 80 and 300 K. - Graphical abstract: Isothermal section of the quasiternary system Ag{sub 2}S–Ga{sub 2}S{sub 3}–In{sub 2}S{sub 3} at 820 K and normalized photoluminescence spectra of the single crystal (Ga{sub 54.59}In{sub 44.66}Er{sub 0.75}){sub 2}S{sub 300} at 300 K. - Highlights: • Isothermal section at 820 K, liquidus surface projection were built for Ag{sub 2}S–Ga{sub 2}S{sub 3}–In{sub 2}S{sub 3}. • Optical properties of single crystals were studied.« less

  11. Comparative Analysis of the Magnitude, Quality, Phenotype and Protective Capacity of SIV Gag-Specific CD8+ T Cells Following Human-, Simian- and Chimpanzee-Derived Recombinant Adenoviral Vector Immunisation

    PubMed Central

    Quinn, Kylie M.; Costa, Andreia Da; Yamamoto, Ayako; Berry, Dana; Lindsay, Ross W.B.; Darrah, Patricia A.; Wang, Lingshu; Cheng, Cheng; Kong, Wing-Pui; Gall, Jason G.D.; Nicosia, Alfredo; Folgori, Antonella; Colloca, Stefano; Cortese, Riccardo; Gostick, Emma; Price, David A.; Gomez, Carmen E.; Esteban, Mariano; Wyatt, Linda S.; Moss, Bernard; Morgan, Cecilia; Roederer, Mario; Bailer, Robert T.; Nabel, Gary J.; Koup, Richard A.; Seder, Robert A.

    2013-01-01

    Recombinant adenoviral vectors (rAds) are the most potent recombinant vaccines for eliciting CD8+ T cell-mediated immunity in humans; however, prior exposure from natural adenoviral infection can decrease such responses. Here we show low seroreactivity in humans against simian- (sAd11, sAd16), or chimpanzee-derived (chAd3, chAd63) compared to human-derived (rAd5, rAd28, rAd35) vectors across multiple geographic regions. We then compared the magnitude, quality, phenotype and protective capacity of CD8+ T cell responses in mice vaccinated with rAds encoding SIV Gag. Using a dose range (1 × 107 to 109 PU), we defined a hierarchy among rAd vectors based on the magnitude and protective capacity of CD8+ T cell responses, from most to least as: rAd5 and chAd3, rAd28 and sAd11, chAd63, sAd16, and rAd35. Selection of rAd vector or dose could modulate the proportion and/or frequency of IFNγ+TNFα+IL-2+ and KLRG1+CD127- CD8+ T cells, but strikingly ~30–80% of memory CD8+ T cells co-expressed CD127 and KLRG1. To further optimise CD8+ T cell responses, we assessed rAds as part of prime-boost regimens. Mice primed with rAds and boosted with NYVAC generated Gag-specific responses that approached ~60% of total CD8+ T cells at peak. Alternatively, priming with DNA or rAd28 and boosting with rAd5 or chAd3 induced robust and equivalent CD8+ T cell responses compared to prime or boost alone. Collectively, these data provide the immunologic basis for using specific rAd vectors alone or as part of prime-boost regimens to induce CD8+ T cells for rapid effector function or robust long-term memory, respectively. PMID:23390298

  12. Adenoviral Delivery of Tumor Necrosis Factor-α and Interleukin-2 Enables Successful Adoptive Cell Therapy of Immunosuppressive Melanoma.

    PubMed

    Siurala, Mikko; Havunen, Riikka; Saha, Dipongkor; Lumen, Dave; Airaksinen, Anu J; Tähtinen, Siri; Cervera-Carrascon, Víctor; Bramante, Simona; Parviainen, Suvi; Vähä-Koskela, Markus; Kanerva, Anna; Hemminki, Akseli

    2016-08-01

    Adoptive T-cell transfer is a promising treatment approach for metastatic cancer, but efficacy in solid tumors has only been achieved with toxic pre- and postconditioning regimens. Thus, adoptive T-cell therapies would benefit from complementary modalities that enable their full potential without excessive toxicity. We aimed to improve the efficacy and safety of adoptive T-cell transfer by using adenoviral vectors for direct delivery of immunomodulatory murine cytokines into B16.OVA melanoma tumors with concomitant T-cell receptor transgenic OT-I T-cell transfer. Armed adenoviruses expressed high local and low systemic levels of cytokine when injected into B16.OVA tumors, suggesting safety of virus-mediated cytokine delivery. Antitumor efficacy was significantly enhanced with adenoviruses coding for murine interleukin-2 (mIL-2) and tumor necrosis factor-α (mTNFα) when compared with T-cell transfer alone or viruses alone. Further improvement in efficacy was achieved with a triple combination of mIL-2, mTNFα, and OT-I T-cells. Mechanistic studies suggest that mIL-2 has an important role in activating T-cells at the tumor, while mTNFα induces chemokine expression. Furthermore, adenovirus treatments enhanced tumor-infiltration of OT-I T-cells as demonstrated by SPECT/CT imaging of (111)In-labeled cells. Our results suggest the utility of cytokine-coding adenoviruses for improving the efficacy of adoptive T-cell therapies.

  13. Adenoviral Delivery of Tumor Necrosis Factor-α and Interleukin-2 Enables Successful Adoptive Cell Therapy of Immunosuppressive Melanoma

    PubMed Central

    Siurala, Mikko; Havunen, Riikka; Saha, Dipongkor; Lumen, Dave; Airaksinen, Anu J.; Tähtinen, Siri; Cervera-Carrascon, Víctor; Bramante, Simona; Parviainen, Suvi; Vähä-Koskela, Markus; Kanerva, Anna; Hemminki, Akseli

    2016-01-01

    Adoptive T-cell transfer is a promising treatment approach for metastatic cancer, but efficacy in solid tumors has only been achieved with toxic pre- and postconditioning regimens. Thus, adoptive T-cell therapies would benefit from complementary modalities that enable their full potential without excessive toxicity. We aimed to improve the efficacy and safety of adoptive T-cell transfer by using adenoviral vectors for direct delivery of immunomodulatory murine cytokines into B16.OVA melanoma tumors with concomitant T-cell receptor transgenic OT-I T-cell transfer. Armed adenoviruses expressed high local and low systemic levels of cytokine when injected into B16.OVA tumors, suggesting safety of virus-mediated cytokine delivery. Antitumor efficacy was significantly enhanced with adenoviruses coding for murine interleukin-2 (mIL-2) and tumor necrosis factor-α (mTNFα) when compared with T-cell transfer alone or viruses alone. Further improvement in efficacy was achieved with a triple combination of mIL-2, mTNFα, and OT-I T-cells. Mechanistic studies suggest that mIL-2 has an important role in activating T-cells at the tumor, while mTNFα induces chemokine expression. Furthermore, adenovirus treatments enhanced tumor-infiltration of OT-I T-cells as demonstrated by SPECT/CT imaging of 111In-labeled cells. Our results suggest the utility of cytokine-coding adenoviruses for improving the efficacy of adoptive T-cell therapies. PMID:27357626

  14. Ordering of calcium and vacancies in calcium catapleiite CaZr[Si{sub 3}O{sub 9}] • 2H{sub 2}O

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

    Aksenov, S. M., E-mail: aks.crys@gmail.com; Portnov, A. M.; Chukanov, N. V.

    A sample of holotypic calcium catapleiite from the Burpala alkaline massif (Northern Baikal, Russia) is studied by single crystal X-ray analysis at 120 K and IR spectroscopy. The empirical formula of calcium catapleiite is Ca{sub 0.97}Na{sub 0.02}Zr{sub 1.01}Si{sub 3}O{sub 9} • 2H{sub 2}O (Z = 4). The X-ray diffraction study confirms the orthorhombic unit cell with the following parameters: a = 7.406(1), b = 12.687(1), and c = 10.112(1) Å; V = 950.1(2) Å{sup 3}; space group Pbnn. The crystal structure is refined in the anisotropic approximation of atomic displacement parameters using 1177 reflections with I > 2σ(I) to themore » final R = 2.91%. The structure of calcium catapleiite under study is based on the microporous heteropolyhedral framework formed by ZrO{sub 6} octahedra and threemembered silicon–oxygen rings [Si{sub 3}O{sub 9}]. It is on the whole analogous to the structures of the samples studied earlier, but differs from them by a high degree of ordering of calcium and vacancies at extraframework positions. The distribution of calcium over Ca1 and Ca2 positions in the calcium catapleiite structure leads to the formation of zigzag chains of the …Ca1–Zr–Ca1–Zr… and …Ca2–h–Ca2–□… types. Low occupancy of the Ca2 position and its alternation with the vacancy are prerequisites for potential Ca{sup 2+} cationic conduction.« less

  15. Bulk and surface half-metallicity: The case of D0{sub 3}-type Mn{sub 3}Ge

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

    Liu, Hao; Gao, G. Y., E-mail: guoying-gao@mail.hust.edu.cn; Hu, Lei

    2014-01-21

    Motivated by the experimental realization of D0{sub 22}-type Mn{sub 3}Ge (001) films [Kurt et al. Appl. Phys. Lett. 101, 132410 (2012)] and the structural stability of D0{sub 3}-type Heusler alloy Mn{sub 3}Ge [Zhang et al. J. Phys.: Condens. Matter 25, 206006 (2013)], we use the first-principles calculations based on the full potential linearized augmented plane-wave method to investigate the electronic and magnetic properties of D0{sub 3}-type Heusler alloy Mn{sub 3}Ge and its (001) surface. We show that bulk D0{sub 3}-Mn{sub 3}Ge is a half-metallic ferromagnet with the minority-spin energy gap of 0.52 eV and the magnetic moment of 1.00 μ{sub B} permore » formula unit. The bulk half-metallicity is preserved at the pure Mn-terminated (001) surface due to the large exchange split, but the MnGe-terminated (001) surface destroys the bulk half-metallicity. We also reveal that the surface stabilities are comparable between the D0{sub 3}-Mn{sub 3}Ge (001) and the experimental D0{sub 22}-Mn{sub 3}Ge (001), which indicates the feasibility to grow the Mn{sub 3}Ge (001) films with D0{sub 3} phase other than D0{sub 22} one. The surface half-metallicity and stability make D0{sub 3}-Mn{sub 3}Ge a promising candidate for spintronic applications.« less

  16. ir-HSP: Improved Recognition of Heat Shock Proteins, Their Families and Sub-types Based On g-Spaced Di-peptide Features and Support Vector Machine

    PubMed Central

    Meher, Prabina K.; Sahu, Tanmaya K.; Gahoi, Shachi; Rao, Atmakuri R.

    2018-01-01

    Heat shock proteins (HSPs) play a pivotal role in cell growth and variability. Since conventional approaches are expensive and voluminous protein sequence information is available in the post-genomic era, development of an automated and accurate computational tool is highly desirable for prediction of HSPs, their families and sub-types. Thus, we propose a computational approach for reliable prediction of all these components in a single framework and with higher accuracy as well. The proposed approach achieved an overall accuracy of ~84% in predicting HSPs, ~97% in predicting six different families of HSPs, and ~94% in predicting four types of DnaJ proteins, with bench mark datasets. The developed approach also achieved higher accuracy as compared to most of the existing approaches. For easy prediction of HSPs by experimental scientists, a user friendly web server ir-HSP is made freely accessible at http://cabgrid.res.in:8080/ir-hsp. The ir-HSP was further evaluated for proteome-wide identification of HSPs by using proteome datasets of eight different species, and ~50% of the predicted HSPs in each species were found to be annotated with InterPro HSP families/domains. Thus, the developed computational method is expected to supplement the currently available approaches for prediction of HSPs, to the extent of their families and sub-types. PMID:29379521

  17. Crystal structures of the ionic conductors Bi{sub 46}M{sub 8}O{sub 89} (M=P, V) related to the fluorite-type structure

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

    Darriet, J.; Launay, J.C.; Zuniga, F.J.

    2005-06-15

    The crystal structures of the two oxides Bi{sub 46}M{sub 8}O{sub 89} (M=P, V) have been solved from single crystals X-ray data at room temperature. Bi{sub 46}P{sub 8}O{sub 89} crystallizes in the monoclinic symmetry (space group C2/m) with the cell parameters a=19.6073(4)A, b=11.4181(4)A, c=21.1119(4)A and {beta}=112.14(3){sup o}. The symmetry of Bi{sub 46}V{sub 8}O{sub 89} is also monoclinic but the space group is P2{sub 1}/c with the unit-cell parameters: a=20.0100(4)A, b=11.6445(4)A, c=20.4136(4)A and {beta}=107.27(3){sup o}. Both structures derive from an oxygen deficient fluorite-type structure where the Bi and M cations (M=P, V) are ordered in the framework. The structures are characterised bymore » isolated MO{sub 4} tetrahedra (M=P, V) which contradicts the previous results. The difference between the two structures is only due to a different order of the M atoms (M=P, V) in the fluorite-type superstructure. It will be shown that some oxygen sites are partially occupied in both structures which can explain the ion conduction properties of these phases. A structural building principle will be proposed that can explain the large domain of solid solution related to the fluorite-type observed in both systems.« less

  18. Hepatic p38α regulates gluconeogenesis by suppressing AMPK.

    PubMed

    Jing, Yanyan; Liu, Wei; Cao, Hongchao; Zhang, Duo; Yao, Xuan; Zhang, Shengjie; Xia, Hongfeng; Li, Dan; Wang, Yu-cheng; Yan, Jun; Hui, Lijian; Ying, Hao

    2015-06-01

    It is proposed that p38 is involved in gluconeogenesis, however, the genetic evidence is lacking and precise mechanisms remain poorly understood. We sought to delineate the role of hepatic p38α in gluconeogenesis during fasting by applying a loss-of-function genetic approach. We examined fasting glucose levels, performed pyruvate tolerance test, imaged G6Pase promoter activity, as well as determined the expression of gluconeogenic genes in mice with a targeted deletion of p38α in liver. Results were confirmed both in vivo and in vitro by using an adenoviral dominant-negative form of p38α (p38α-AF) and the constitutively active mitogen-activated protein kinase 6, respectively. Adenoviral dominant-negative form of AMP-activated protein kinase α (DN-AMPKα) was employed to test our proposed model. Mice lacking hepatic p38α exhibited reduced fasting glucose level and impaired gluconeogenesis. Interestingly, hepatic deficiency of p38α did not result in an alteration in CREB phosphorylation, but led to an increase in AMPKα phosphorylation. Adenoviral DN-AMPKα could abolish the effect of p38α-AF on gluconeogenesis. Knockdown of up-steam transforming growth factor β-activated kinase 1 decreased the AMPKα phosphorylation induced by p38α-AF, suggesting a negative feedback loop. Consistently, inverse correlations between p38 and AMPKα phosphorylation were observed during fasting and in diabetic mouse models. Importantly, adenoviral p38α-AF treatment ameliorated hyperglycemia in diabetic mice. Our study provides evidence that hepatic p38α functions as a negative regulator of AMPK signaling in maintaining gluconeogenesis, dysregulation of this regulatory network contributes to unrestrained gluconeogenesis in diabetes, and hepatic p38α could be a drug target for hyperglycemia. Copyright © 2015 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

  19. Matching organic libraries with protein-substructures

    NASA Astrophysics Data System (ADS)

    Preissner, R.; Goede, A.; Rother, K.; Osterkamp, F.; Koert, U.; Froemmel, C.

    2001-09-01

    We present a general approach which allows automatic identification of sub-structures in proteins that resemble given three-dimensional templates. This paper documents its success with non-peptide templates such as β-turn mimetics. We considered well-tested turn-mimetics such as the bicyclic turned dipeptide (BTD), spiro lactam (Spiro) and the 2,5-disubstituded tetrahydrofuran (THF), a new furan-derivative which was recently developed and characterized. The detected geometric similarity between the templates and the protein patches corresponds to r.m.s.-values of 0.3 Å for more than 80% of the constituting atoms, which is typical for active site comparisons of homologous proteins. This fast automatic procedure might be of biomedical value for finding special mimicking leads for particular protein sub-structures as well as for template-assembled synthetic protein (TASP) design.

  20. The Effect of Brief Exposure to Sub-Therapeutic Concentrations of Chlorhexidine Digluconate on the Susceptibility of Staphylococci to Platelet Microbicidal Protein.

    PubMed

    Ivanov, Iuri B; Gritsenko, Viktor A; Kuzmin, Michael D

    2015-06-01

    Antiseptic agents are widely used in hospitals and are essential when prevention and control of nosocomial infections is required. It is necessary to consider several aspects that affect the biocide activity because they have direct impact on the nosocomial infection rate. Organisms belonging to the Staphylococcus genus are involved in such infections and chlorhexidine digluconate (CHXD) is one of the most used antiseptic agents for human and animal health. In the context of such infections, anti-bacterial peptides have been isolated from platelets and have been termed platelet microbicidal proteins (PMP). Platelet microbicidal proteins have been shown to enhance the bacterial inhibitory activities of sub-therapeutic concentrations of antibiotics. The main objective of this study was to investigate the effect of brief exposure to different sub-therapeutic concentrations of CHXD on the susceptibility of staphylococci to PMP. The influence of brief exposure to three different sub-therapeutic concentrations of CHXD (0.005%, 0.0025%, and 0.00125%) on the subsequent staphylocidal effect of PMP was evaluated. Among all clinical staphylococcal strains studied, all isolates were considered to be resistant to the bactericidal action of PMP. Exposure of staphylococci to CHXD prior to PMP resulted in significantly increased staphylococcal killing compared with the killing achieved with PMP alone. This enhanced effect was most marked for concentrations of CHXD of 0.005%. The combined data indicate that PMP exerts cooperative bactericidal effect with CHXD. The anti-staphylococcal PMP and CHXD synergistic activity in vitro demonstrated in the present study make these molecules potentially useful for preventing endovascular catheter-associated infections. Future research based on animal and human models is needed to elucidate the in vivo efficacies and toxicities and utility in clinical practice.

  1. Variation of k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} for the small-field dosimetric parameters percentage depth dose, tissue-maximum ratio, and off-axis ratio

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

    Francescon, Paolo, E-mail: paolo.francescon@ulssvicenza.it; Satariano, Ninfa; Beddar, Sam

    Purpose: Evaluate the ability of different dosimeters to correctly measure the dosimetric parameters percentage depth dose (PDD), tissue-maximum ratio (TMR), and off-axis ratio (OAR) in water for small fields. Methods: Monte Carlo (MC) simulations were used to estimate the variation of k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}} for several types of microdetectors as a function of depth and distance from the central axis for PDD, TMR, and OAR measurements. The variation of k{sub Q{sub c{sub l{sub i{sub n,Q{sub m{sub s{sub r}{sup f{sub c}{sub l}{sub i}{sub n},f{sub m}{sub s}{sub r}}}}}}}}}more » enables one to evaluate the ability of a detector to reproduce the PDD, TMR, and OAR in water and consequently determine whether it is necessary to apply correction factors. The correctness of the simulations was verified by assessing the ratios between the PDDs and OARs of 5- and 25-mm circular collimators used with a linear accelerator measured with two different types of dosimeters (the PTW 60012 diode and PTW PinPoint 31014 microchamber) and the PDDs and the OARs measured with the Exradin W1 plastic scintillator detector (PSD) and comparing those ratios with the corresponding ratios predicted by the MC simulations. Results: MC simulations reproduced results with acceptable accuracy compared to the experimental results; therefore, MC simulations can be used to successfully predict the behavior of different dosimeters in small fields. The Exradin W1 PSD was the only dosimeter that reproduced the PDDs, TMRs, and OARs in water with high accuracy. With the exception of the EDGE diode, the stereotactic diodes reproduced the PDDs and the TMRs in water with a systematic error of less than 2% at depths of up to 25 cm; however, they produced OAR values that were significantly different from those in water, especially in the tail region (lower than 20% in some cases). The microchambers could be used for PDD

  2. The effect of pre-existing malnutrition on pediatric burn mortality in a sub-Saharan African burn unit.

    PubMed

    Grudziak, Joanna; Snock, Carolyn; Mjuweni, Stephen; Gallaher, Jared; Cairns, Bruce; Charles, Anthony

    2017-11-01

    Nutritional status predicts burn outcomes in the developed world, but its effect on burn mortality in the developing world has not been widely studied. In sub Saharan Africa, burn is primarily a disease of children, and the majority of children in sub-Saharan Africa are malnourished. We therefore sought to determine the prevalence and effect of malnutrition on burn mortality at our institution. This is a retrospective review of children aged 0-5, with anthropomorphic measurements available, who were admitted to our burn unit from July 2011 to May 2016. Age-adjusted Z scores were calculated for height, weight, weight for height, and mid-upper arm circumference (MUAC). Following bivariate analysis, we used logistic regression to construct a fully adjusted model of predictors of mortality. Of the 1357 admitted patients, 839 (61.2%) were aged 0-5. Of those, 512 (62.9%) had one or more anthropomorphic measurements available, and were included in the analysis. 54% were male, and the median age was 28 months. The median TBSA was 15%, with a majority of burns caused by scalds (77%). Mortality was 16%. Average Z-score for any of the indicators of malnutrition was -1.45±1.66. TBSA (OR: 1.08, 95% CI: 1.06, 1.11), decreasing Z-score (OR: 1.19, 95% CI: 1.00, 1.41), and flame burn (OR: 2.51, 95% CI: 1.40, 4.49) were associated with an increase in mortality. Preexisting malnutrition in burn patients in sub-Saharan Africa increases odds of mortality after controlling for significant covariates. Survival of burn patients in this region will not reach that of the developed world until a strategy of aggressive nutritional support is implemented in this population. Copyright © 2017 Elsevier Ltd and ISBI. All rights reserved.

  3. The Inner Membrane Complex Sub-compartment Proteins Critical for Replication of the Apicomplexan Parasite Toxoplasma gondii Adopt a Pleckstrin Homology Fold*

    PubMed Central

    Tonkin, Michelle L.; Beck, Josh R.; Bradley, Peter J.; Boulanger, Martin J.

    2014-01-01

    Toxoplasma gondii, an apicomplexan parasite prevalent in developed nations, infects up to one-third of the human population. The success of this parasite depends on several unique structures including an inner membrane complex (IMC) that lines the interior of the plasma membrane and contains proteins important for gliding motility and replication. Of these proteins, the IMC sub-compartment proteins (ISPs) have recently been shown to play a role in asexual T. gondii daughter cell formation, yet the mechanism is unknown. Complicating mechanistic characterization of the ISPs is a lack of sequence identity with proteins of known structure or function. In support of elucidating the function of ISPs, we first determined the crystal structures of representative members TgISP1 and TgISP3 to a resolution of 2.10 and 2.32 Å, respectively. Structural analysis revealed that both ISPs adopt a pleckstrin homology fold often associated with phospholipid binding or protein-protein interactions. Substitution of basic for hydrophobic residues in the region that overlays with phospholipid binding in related pleckstrin homology domains, however, suggests that ISPs do not retain phospholipid binding activity. Consistent with this observation, biochemical assays revealed no phospholipid binding activity. Interestingly, mapping of conserved surface residues combined with crystal packing analysis indicates that TgISPs have functionally repurposed the phospholipid-binding site likely to coordinate protein partners. Recruitment of larger protein complexes may also be aided through avidity-enhanced interactions resulting from multimerization of the ISPs. Overall, we propose a model where TgISPs recruit protein partners to the IMC to ensure correct progression of daughter cell formation. PMID:24675080

  4. Floral ontogeny and gene protein localization rules out euanthial interpretation of reproductive units in Lepironia (Cyperaceae, Mapanioideae, Chrysitricheae)

    PubMed Central

    Prychid, C. J.; Bruhl, J. J.

    2013-01-01

    Background and Aims In the sedge subfamily Mapanioideae there are considerable discrepancies between the standard trimerous monocot floral architecture expected and the complex floral and inflorescence morphologies seen. Decades of debate about whether the basic reproductive units are single flowers or pseudanthia have not resolved the question. This paper evaluates current knowledge about Mapaniid reproductive structures and presents an ontogenetic study of the Mapaniid genus Lepironia with the first floral protein expression maps for the family, localizing the products of the APETALA1/FRUITFULL-like (AP1/FUL) MADS-box genes with the aim of shedding light on this conundrum. Methods A range of reproductive developmental stages, from spikelet primordia through to infructescence material, were processed for anatomical and immunohistochemical analyses. Key Results The basic reproductive unit is subtended by a bract and possesses two prophyll-like structures, the first organs to be initiated on the primordium, which grow rapidly, enclosing two whorls of initiating leaf-like structures with intervening stamens and a central gynoecium, formed from an annular primordium. The subtending bract and prophyll-like structures possess very different morphologies from that of the internal leaf-like structures and do not show AP1/FUL-like protein localization, which is otherwise strongly localized in the internal leaf-like structures, stamens and gynoecia. Conclusions Results support the synanthial hypothesis as the evolutionary origin of the reproductive unit. Thus, the basic reproductive unit in Lepironia is an extremely condensed pseudanthium, of staminate flowers surrounding a central terminal pistillate female flower. Early in development the reproductive unit becomes enclosed by a split-prophyll, with the whole structure subtended by a bract. PMID:23723258

  5. Angiotensin II type 1 receptor-associated protein regulates carotid intimal hyperplasia through controlling apoptosis of vascular smooth muscle cells.

    PubMed

    Yue, Yongqiang; Ma, Ke; Li, Zhen; Wang, Zhonggao

    2018-01-08

    Intimal hyperplasia is the main cause of restenosis after carotid artery injury, and the underlying mechanism involves the proliferation and migration of vascular smooth muscle cells (VSMCs). Angiotensin II Type 1 Receptor-Associated Protein (ATRAP) has been reported to withstand intimal hyperplasia by inhibiting VSMCs proliferation and migration; however, whether the beneficial effect of ATRAP associates with VSMCs apoptosis remains unclarified. We demonstrated that the adenoviral-mediated overexpression of ATRAP induced VSMC apoptosis, alleviating the balloon injury-induced neointima formation in rats. Under the condition of Angiotensin-II stimulation, ATRAP overexpression induced the apoptosis of rat VSMCs by depressing the PI3K-Akt signaling; whereas up-regulation of Akt by PTEN inhibitor abolished the apoptotic death. Thus, ATRAP regulates carotid intimal hyperplasia through controlling the PI3K-Akt signal-mediated VSMCs apoptosis. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Magnetic properties of (SrFe{sub 12}O{sub 19}){sub x}(CaCu{sub 3}Ti{sub 4}O{sub 12}){sub 1–x} composites

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

    Eremina, R. M., E-mail: REremina@yandex.ru; Sharipov, K. R.; Yatsyk, I. V.

    2016-07-15

    New composite materials (SrFe{sub 12}O{sub 19}){sub x}(CaCu{sub 3}Ti{sub 4}O{sub 12}){sub 1–x} (x = 0, 0.05, 1) have been synthesized. Their magnetic properties are studied in the temperature range 5–300 K using the magnetic resonance and magnetometry methods. It is found that strontium hexaferrite microinclusions in the (SrFe{sub 12}O{sub 19}){sub 0.05}(CaCu{sub 3}Ti{sub 4}O{sub 12}){sub 0.95} composite “magnetize” CaCu{sub 3}Ti{sub 4}O{sub 12} at temperatures from 300 to 200 K, forming a ferrimagnetic particle near the SrFe{sub 12}O{sub 19} “core.” The magnetic resonance line below 200 K splits into two lines corresponding to SrFe{sub 12}O{sub 19} and CaCu{sub 3}Ti{sub 4}O{sub 12}. The coremore » effect decoration is manifested in the increase in the Curie–Weiss temperature from 25 K in CaCu{sub 3}Ti{sub 4}O{sub 12} without the doping ceramics to 80 K in the composite with 5% of SrFe{sub 12}O{sub 19}.« less

  7. “Ni{sub 5}TiO{sub 7}” is Ni{sub 5}TiO{sub 4}(BO{sub 3}){sub 2}

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

    Nalbandyan, V.B.

    2017-05-15

    It is shown that the compound known as Ni{sub 5}TiO{sub 7} and considered as a promising catalyst and oxidation product of alloys does not exist and its XRD pattern actually corresponds to Ni{sub 5}TiO{sub 4}(BO{sub 3}){sub 2} - Graphical abstract: XRD pattern of “Ni{sub 5}TiO{sub 7}” (top) is identical to that for Ni{sub 5}TiO{sub 4}(BO{sub 3}){sub 2} (bottom) based on single-crystal structural data. - Highlights: • Popular catalyst known as Ni{sub 5}TiO{sub 7} is actually Ni{sub 5}TiO{sub 4}(BO{sub 3}){sub 2}. • B{sub 2}O{sub 3} came from the flux used for crystal growth. • Some authors reporting this phase did notmore » use any boron compounds.« less

  8. Pilot plant studies of the CO{sub 2} capture performance of aqueous MEA and mixed MEA/MDEA solvents at the University of Regina CO{sub 2} capture technology development plant and the Boundary Dam CO{sub 2} capture demonstration

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

    Idem, R.; Wilson, M.; Tontiwachwuthikul, P.

    2006-04-12

    Evaluations of the benefits of using a mixed MEA/MDEA solvent for CO{sub 2} capture in terms of the heat requirement for solvent regeneration, lean and rich loadings, CO{sub 2} production, and solvent stability were performed by comparing the performance of aqueous 5 kmol/m{sup 3} MEA with that of an aqueous 4:1 molar ratio MEA/MDEA blend of 5 kmol/ml total amine concentration as a function of the operating time. The tests were performed using two pilot CO{sub 2} capture plants of the International Test Centre for CO{sub 2} Capture (ITC), which provided two different sources and compositions of flue gas. Themore » University of Regina CO{sub 2} plant (UR unit) processes flue gas from the combustion of natural gas while the Boundary Dam CO{sub 2} plant (BD unit) processes flue gas from a coal-fired electric power station. The results show that a huge heat-duty reduction can be achieved by using a mixed MEA/MDEA solution instead of a single MEA solution in an industrial environment of a CO{sub 2} capture plant. However, this benefit is dependent on whether the chemical stability of the solvent can be maintained.« less

  9. New insights into the application of the valence rules in Zintl phases—Crystal and electronic structures of Ba{sub 7}Ga{sub 4}P{sub 9}, Ba{sub 7}Ga{sub 4}As{sub 9}, Ba{sub 7}Al{sub 4}Sb{sub 9}, Ba{sub 6}CaAl{sub 4}Sb{sub 9}, and Ba{sub 6}CaGa{sub 4}Sb{sub 9}

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

    He, Hua; Stoyko, Stanislav; Bobev, Svilen, E-mail: bobev@udel.edu

    Crystals of three new ternary pnictides—Ba{sub 7}Al{sub 4}Sb{sub 9}, Ba{sub 7}Ga{sub 4}P{sub 9}, and Ba{sub 7}Ga{sub 4}As{sub 9} have been prepared by reactions of the respective elements in molten Al or Pb fluxes. Single-crystal X-ray diffraction studies reveal that the three phases are isotypic, crystallizing in the orthorhombic Ba{sub 7}Ga{sub 4}Sb{sub 9}-type structure (space group Pmmn, Pearson symbol oP40, Z=2), for which only the prototype is known. The structure is based on TrPn{sub 4} tetrahedra (Tr=Al, Ga; Pn=P, As, Sb), connected in an intricate scheme into 1D-ribbons. Long interchain Pn–Pn bonds (d{sub P–P}>3.0 Å; d{sub As–As}>3.1 Å; d{sub Sb–Sb}>3.3 Å)more » account for the realization of 2D-layers, separated by Ba{sup 2+} cations. Applying the classic valance rules to rationalize the bonding apparently fails, and Ba{sub 7}Ga{sub 4}Sb{sub 9} has long been known as a metallic Zintl phase. Earlier theoretical calculations, both empirical and ab-initio, suggest that the possible metallic properties originate from filled anti-bonding Pn–Pn states, and the special roles of the “cations” in this crystal structure. To experimentally probe this hypothesis, we sought to synthesize the ordered quaternary phases Ba{sub 6}CaTr{sub 4}Sb{sub 9} (Tr=Al, Ga). Single-crystal X-ray diffraction work confirms Ba{sub 6.145(3)}Ca{sub 0.855}Al{sub 4}Sb{sub 9} and Ba{sub 6.235(3)}Ca{sub 0.765}Ga{sub 4}Sb{sub 9}, with Ca atoms preferably substituting Ba on one of the three available sites. The nuances of the five crystal structures are discussed, and the chemical bonding in Ba{sub 7}Ga{sub 4}As{sub 9} is interrogated by tight-binding linear muffin-tin orbital calculations. - Graphical abstract: The new Zintl phases—Ba{sub 7}Al{sub 4}Sb{sub 9}, Ba{sub 7}Ga{sub 4}P{sub 9}, and Ba{sub 7}Ga{sub 4}As{sub 9}, and their quaternary variants Ba{sub 6}CaTr{sub 4}Sb{sub 9} (Tr=Al, Ga)—crystallize in the Ba{sub 7}Ga{sub 4}Sb{sub 9} structure type. The structures

  10. Phase equilibria in the quasi-ternary system Ag{sub 2}Se–Ga{sub 2}Se{sub 3}–In{sub 2}Se{sub 3} and physical properties of (Ga{sub 0.6}In{sub 0.4}){sub 2}Se{sub 3}, (Ga{sub 0.594}In{sub 0.396}Er{sub 0.01}){sub 2}Se{sub 3} single crystals

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

    Ivashchenko, I.A., E-mail: inna.ivashchenko@mail.ru; Danyliuk, I.V.; Olekseyuk, I.D.

    2014-02-15

    The quasi-ternary system Ag{sub 2}Se–Ga{sub 2}Se{sub 3}–In{sub 2}Se{sub 3} was investigated by differential thermal, X-ray phase, X-ray structure, microstructure analysis and microhardness measurements. Five quasi-binary phase diagrams, six polythermal sections, isothermal section at 820 K and the liquidus surface projection were constructed. The character and temperature of the invariant processes were determined. The specific resistance of the single crystals (Ga{sub 0.6}In{sub 0.4}){sub 2}Se{sub 3}, (Ga{sub 0.594}In{sub 0.396}Er{sub 0.01}){sub 2}Se{sub 3} was measured, 7.5×10{sup 5} and 3.15×10{sup 5} Ω m, respectively, optical absorption spectra in the 600–1050 nm range were recorded at room temperature, and the band gap energy was estimatedmore » which is 1.95±0. 01 eV for both samples. - Graphical abstract: The article reports for the first time the investigated liquidus surface projection of the Ag{sub 2}Se–Ga{sub 2}Se{sub 3}–In{sub 2}Se{sub 3} system and isothermal section at 820 K of the system. Five phase diagrams, six polythermal sections, isothermal section at 820 K and the liquidus surface projection were built at the first time. The existence of the large region of the solid solutions based on AgIn{sub 5}Se{sub 8}, Ga{sub 2}Se{sub 3} and AgGa{sub 1−x}In{sub x}Se{sub 2} was investigated. The existence of two ternary phases was established in the Ga{sub 2}Se{sub 3}–In{sub 2}Se{sub 3} system. Two single crystals (Ga{sub 0.6}In{sub 0.4}){sub 2}Se{sub 3}, (Ga{sub 0.594}In{sub 0.396}Er{sub 0.01}){sub 2}Se{sub 3} were grown and some of optical properties of them were studied at first time. Display Omitted - Highlights: • Liquidus surface projection was built for Ag{sub 2}Se–Ga{sub 2}Se{sub 3}–In{sub 2}Se{sub 3} system. • Solid solution ranges of AgIn{sub 5}Se{sub 8}, Ga{sub 2}Se{sub 3} and AgGa{sub 1−x}In{sub x}Se{sub 2} were investigated. • Two single crystals (Ga{sub 0.6}In{sub 0.4}){sub 2}Se{sub 3}, (Ga{sub 0.594}In{sub 0

  11. Magnetic properties of quadruple perovskite solid solutions Ca{sub 1–x}Y{sub x}Cu{sub 3}Fe{sub 4}O{sub 12} and Y{sub 1–y}Ce{sub x}Cu{sub 3}Fe{sub 4}O{sub 12}

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

    Murakami, Makoto; Mori, Shigeo; Yamada, Ikuya, E-mail: i-yamada@21c.osakafu-u.ac.jp

    Magnetic properties of the quadruple perovskite solid solutions Ca{sub 1–x}Y{sub x}Cu{sub 3}Fe{sub 4}O{sub 12} and Y{sub 1–y}Ce{sub y}Cu{sub 3}Fe{sub 4}O{sub 12} are investigated. Ca{sub 1–x}Y{sub x}Cu{sub 3}Fe{sub 4}O{sub 12} shows continuous increase in the ferromagnetic transition temperature as x increases. Y{sub 1–y}Ce{sub y}Cu{sub 3}Fe{sub 4}O{sub 12} exhibits a ferromagnetic-antiferromagnetic transition in the vicinity of y = 0.5. These observations demonstrate the electron doping effect on magnetic properties of charge-disproportionated ACu{sub 3}Fe{sub 4}O{sub 12} phases.

  12. Cell segmentation in time-lapse fluorescence microscopy with temporally varying sub-cellular fusion protein patterns.

    PubMed

    Bunyak, Filiz; Palaniappan, Kannappan; Chagin, Vadim; Cardoso, M

    2009-01-01

    Fluorescently tagged proteins such as GFP-PCNA produce rich dynamically varying textural patterns of foci distributed in the nucleus. This enables the behavioral study of sub-cellular structures during different phases of the cell cycle. The varying punctuate patterns of fluorescence, drastic changes in SNR, shape and position during mitosis and abundance of touching cells, however, require more sophisticated algorithms for reliable automatic cell segmentation and lineage analysis. Since the cell nuclei are non-uniform in appearance, a distribution-based modeling of foreground classes is essential. The recently proposed graph partitioning active contours (GPAC) algorithm supports region descriptors and flexible distance metrics. We extend GPAC for fluorescence-based cell segmentation using regional density functions and dramatically improve its efficiency for segmentation from O(N(4)) to O(N(2)), for an image with N(2) pixels, making it practical and scalable for high throughput microscopy imaging studies.

  13. Fluorescence Studies of Protein Crystal Nucleation

    NASA Technical Reports Server (NTRS)

    Pusey, Marc; Sumida, John

    2000-01-01

    We have postulated that, in the case of tetragonal chicken egg white lysozyme, crystal growth occurs by the addition of pre-critical nuclei sized n-mers that form in the bulk solution, and that the n-mer growth units were multiples of the tetrameric 4(sub 3) helical structure. These have the strongest intermolecular bonds in the crystal and are therefore likely to be the first species formed. High resolution AFM studies provide strong supporting evidence for this model, but the data also suggest that the actual species in solution may not be identical in structure to that found in the crystal. We are using fluorescence resonance energy transfer (FRET) to study the initial solution phase self-assembly process, using covalent fluorescent derivatives which crystallize in the characteristic P4(sub 3)2(sub 1)2(sub 1) space group. FRET studies are being carried out between the cascade blue (CB-lys, donor, Ex(sub max) 366 nm, Em 420 nm) and lucifer yellow (LY-lys, acceptor, Ex(sub max) 430 nm, Em 528 nm) asp101 derivatives. The estimated R(sub 0) for this probe pair, the distance where 50% of the donor energy is transferred to the acceptor, is approx. 1.2 nm, compared to 2.2 nm between the side chain carboxyls of adjacent asp101's in the crystalline 4(sub 3) helix. The short donor lifetime of 2.80 ns (chi(sup 2) = 0.644), coupled with the large average distances between the molecules (greater than or equal to 50 nm) in solution, ensure that any energy transfer observed is not due to random diffusive interactions. Lifetime data show that CB-lys has a single lifetime when it is the only species in solution. Similarly, LY-lys also exhibits a single lifetime of 4.63 ns (chi(sup 2) = 0.42) when alone in solution. Addition of LY-lys to CB-lys results in the appearance of a third lifetime component of 0.348ns for the CB-lys. The fractional intensities of the different species present can be used to estimate the distribution of monomer and n-mers in solution. The self

  14. Neutron diffraction studies of the Na-ion battery electrode materials NaCoCr{sub 2}(PO{sub 4}){sub 3}, NaNiCr{sub 2}(PO{sub 4}){sub 3}, and Na{sub 2}Ni{sub 2}Cr(PO{sub 4}){sub 3}

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

    Yahia, H. Ben; Essehli, R., E-mail: ressehli@qf.org.qa; Avdeev, M.

    The new compounds NaCoCr{sub 2}(PO{sub 4}){sub 3}, NaNiCr{sub 2}(PO{sub 4}){sub 3}, and Na{sub 2}Ni{sub 2}Cr(PO{sub 4}){sub 3} were synthesized by sol-gel method and their crystal structures were determined by using neutron powder diffraction data. These compounds were characterized by galvanometric cycling and cyclic voltammetry. NaCoCr{sub 2}(PO{sub 4}){sub 3}, NaNiCr{sub 2}(PO{sub 4}){sub 3}, and Na{sub 2}Ni{sub 2}Cr(PO{sub 4}){sub 3} crystallize with a stuffed α-CrPO{sub 4}-type structure. The structure consists of a 3D-framework made of octahedra and tetrahedra that are sharing corners and/or edges generating channels along [100] and [010], in which the sodium atoms are located. Of significance, in the structuresmore » of NaNiCr{sub 2}(PO{sub 4}){sub 3}, and Na{sub 2}Ni{sub 2}Cr(PO{sub 4}){sub 3} a statistical disorder Ni{sup 2+}/Cr{sup 3+} was observed on both the 8g and 4a atomic positions, whereas in NaCoCr{sub 2}(PO{sub 4}){sub 3} the statistical disorder Co{sup 2+}/Cr{sup 3+} was only observed on the 8g atomic position. When tested as negative electrode materials, NaCoCr{sub 2}(PO{sub 4}){sub 3}, NaNiCr{sub 2}(PO{sub 4}){sub 3}, and Na{sub 2}Ni{sub 2}Cr(PO{sub 4}){sub 3} delivered specific capacities of 352, 385, and 368 mA h g{sup −1}, respectively, which attests to the electrochemical activity of sodium in these compounds. - Highlights: • NaCoCr{sub 2}(PO{sub 4}){sub 3}, NaNiCr{sub 2}(PO{sub 4}){sub 3}, and Na{sub 2}Ni{sub 2}Cr(PO{sub 4}){sub 3} were synthesized by sol-gel method. • The crystal structures were determined by using neutron powder diffraction data. • The three compounds crystallize with a stuffed α-CrPO{sub 4}-type structure. • The three compounds were tested as anodes in sodium-ion batteries. • Relatively high specific capacities were obtained for these compounds.« less

  15. The new Zintl phases Eu{sub 21}Cd{sub 4}Sb{sub 18} and Eu{sub 21}Mn{sub 4}Sb{sub 18}

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

    Wang, Yi; Darone, Gregory M.; Bobev, Svilen, E-mail: bobev@udel.edu

    Crystals of two new Zintl compounds, Eu{sub 21}Mn{sub 4}Sb{sub 18} and Eu{sub 21}Cd{sub 4}Sb{sub 18} have been synthesized using the molten metal flux method, and their structures have been established by single-crystal X-ray diffraction. Both compounds are isotypic and crystallize in the monoclinic space group C2/m (No. 12, Z=4). The structures are based on edge- and corner-shared MnSb{sub 4} or CdSb{sub 4} tetrahedra, which make octameric [Mn{sub 8}Sb{sub 22}] or [Cd{sub 8}Sb{sub 22}] polyanions. Homoatomic Sb–Sb bonds are present in both structures. The Eu atoms take the role of Eu{sup 2+}cations with seven unpaired 4f electrons, as suggested by themore » temperature-dependent magnetization measurements. The magnetic susceptibilities of Eu{sub 21}Mn{sub 4}Sb{sub 18} and Eu{sub 21}Cd{sub 4}Sb{sub 18} indicate that both phases order anti-ferromagnetically with Néel temperatures of ca. 7 K and ca. 10 K, respectively. The unpaired 3d electrons of the Mn atoms in Eu{sub 21}Mn{sub 4}Sb{sub 18} do contribute to the magnetic response, however, the bulk magnetization measurements do not provide evidence for long-range ordering of the Mn spins down to 5 K. Electrical resistivity measurements suggest that both compounds are narrow band gap semiconductors. - Graphical abstract: Eu{sub 21}Mn{sub 4}Sb{sub 18} and Eu{sub 21}Cd{sub 4}Sb{sub 18} have complex monoclinic structures, based on MnSb{sub 4} and CdSb{sub 4} tetrahedra, both edge- and corner-shared. A perspective of the crystal structure is shown, as viewed along the b axis. Display Omitted - Highlights: • Eu{sub 21}Mn{sub 4}Sb{sub 18} and Eu{sub 21}Cd{sub 4}Sb{sub 18} are novel compounds in the respective ternary phase diagrams. • For both structures, the Zintl-Klemm rules are followed, and both are small gap semiconductors. • Eu{sub 21}Mn{sub 4}Sb{sub 18} and Eu{sub 21}Cd{sub 4}Sb{sub 18} are air-stable Zintl phases and could be new thermoelectric materials.« less

  16. Phase diagram of the Y–Y{sub 2}Se{sub 3} system, enthalpies of phase transformations

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

    Andreev, O.V.; Kharitontsev, V.B.; Polkovnikov, A.A., E-mail: a.a.polkovnikov@yandex.ru

    A phase diagram for the Y–Y{sub 2}Se{sub 3} system has been constructed in which the YSe and Y{sub 2}Se{sub 3} phases melt congruently. The daltonide type YSe phase (ST Y{sub 0,75}Se, a=1.1393 nm, melting point=2380 K, H=2200 MPa) forms a double-sided solid solution from 49–50–53 at% Se. In the 50–53 at% Se range, the unit cell parameter increases to 1.1500 nm, the microhardness increases to 4100 MPa and electrical resistivity increases from 0.018 to 0.114 Ω m. These changes are caused by the dominating influx of newly formed structural cationic vacancies arising from the selenium anions that are surplus formore » the 1:1 Y:Se stoichiometry. The full-valence Y{sub 2}Se{sub 3} composition exists as a low-temperature modification of ε-Y{sub 2}Se{sub 3} (ST Sc{sub 2}S{sub 3}, a=1.145 nm, b=0.818 nm, c=2.438 nm, melting point=1780 K, ∆fusion enthalpy=4±0.4 J/g) and transforms into a modification of ξ-Y{sub 2}Se{sub 3} that does not undergo fixing by thermo-hardening. The eutectic melting point between the YSe and Y{sub 2}Se{sub 3} phases is 1625±5 K, with a eutectic composition that is assumed to be 57.5 at% Se and have an enthalpy of fusion of 43±4.3 J/g. The eutectic for the Y and YSe phases appears at a temperature of 1600 K and 5 at% Se. - Highlights: • Phase equilibria in the Y–Y{sub 2}Se{sub 3} system from 1000 K to melt were studies. • High-temperature polymorphic transition for Y{sub 2}Se{sub 3} were observed. • Singular points in solid solutions areas for YSe and Y{sub 2}Se{sub 3} were found.« less

  17. Universal partitioning of the hierarchical fold network of 50-residue segments in proteins

    PubMed Central

    Ito, Jun-ichi; Sonobe, Yuki; Ikeda, Kazuyoshi; Tomii, Kentaro; Higo, Junichi

    2009-01-01

    Background Several studies have demonstrated that protein fold space is structured hierarchically and that power-law statistics are satisfied in relation between the numbers of protein families and protein folds (or superfamilies). We examined the internal structure and statistics in the fold space of 50 amino-acid residue segments taken from various protein folds. We used inter-residue contact patterns to measure the tertiary structural similarity among segments. Using this similarity measure, the segments were classified into a number (Kc) of clusters. We examined various Kc values for the clustering. The special resolution to differentiate the segment tertiary structures increases with increasing Kc. Furthermore, we constructed networks by linking structurally similar clusters. Results The network was partitioned persistently into four regions for Kc ≥ 1000. This main partitioning is consistent with results of earlier studies, where similar partitioning was reported in classifying protein domain structures. Furthermore, the network was partitioned naturally into several dozens of sub-networks (i.e., communities). Therefore, intra-sub-network clusters were mutually connected with numerous links, although inter-sub-network ones were rarely done with few links. For Kc ≥ 1000, the major sub-networks were about 40; the contents of the major sub-networks were conserved. This sub-partitioning is a novel finding, suggesting that the network is structured hierarchically: Segments construct a cluster, clusters form a sub-network, and sub-networks constitute a region. Additionally, the network was characterized by non-power-law statistics, which is also a novel finding. Conclusion Main findings are: (1) The universe of 50 residue segments found here was characterized by non-power-law statistics. Therefore, the universe differs from those ever reported for the protein domains. (2) The 50-residue segments were partitioned persistently and universally into some dozens (ca

  18. Structural and optical properties of the naked and passivated Al{sub 5}Au{sub 5} bimetallic nanoclusters

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

    Grande-Aztatzi, Rafael; Formoso, Elena; Matxain, Jon M.

    The structural and optical properties of both the naked and passivated bimetallic Al{sub 5}Au{sub 5} nanoclusters have been analyzed based on data obtained from ab initio density functional theory and quantum molecular dynamics simulations. It has been found that the Al{sub 5}Au{sub 5} nanocluster possesses a hollow shaped minimum energy structure with segregated Al and Au layered domains, the former representing the electrophilic domain and the latter the nucleophilic domain. In particular, it has been shown that alkali metal cations attach in the nucleophilic domain and hop from one Au site to the next one in the picoseconds time scale,more » while anions are bound tightly to the Al atoms of the electrophilic domain. Simulating annealing studies are very suggestive of the proneness of the nanocluster towards coalescence into large cluster units, when the cluster is left unprotected by appropriate ligands. Further passivation studies with NaF salt suggest, nonetheless, the possibility of the isolation of the Al{sub 5}Au{sub 5} cluster in molten salts or ionic liquids.« less

  19. Estimation of the protein content of US imports of milk protein concentrates.

    PubMed

    Bailey, K W

    2003-12-01

    Recent declines in milk prices in the United States have sparked renewed concern that imports of milk protein concentrates (MPC) are increasingly entering the United States with very low tariff rates and is having an adverse impact on the US dairy industry. Milk protein concentrates are used in the United States in many different products, including the starter culture of cheese, or in nonstandard cheeses such as baker's cheese, ricotta, Feta and Hispanic cheese, processed cheese foods, and nutritional products. One of the difficult aspects of trying to assess the impact of MPC imports on the US dairy industry is to quantify the protein content of these imports. The protein content of MPC imports typically ranges from 40 to 88%. The purpose of this study is to develop a methodology that can be used to estimate the protein content of MPC on a country by country basis. Such an estimate would not only provide information regarding the quantity of protein entering the United States, but would also provide a profile of low- and high-value MPC importers. This is critical for market analysis, since it is the lower valued MPC imports that more directly displaces US-produced skim milk powder.

  20. Impaired Expression of Uncoupling Protein 2 (UCP2) Causes Defective Post-ischemic Angiogenesis in Mice Deficient in AMP-activated Protein Kinase α Subunits

    PubMed Central

    Xu, Ming-Jiang; Song, Ping; Shirwany, Najeeb; Liang, Bin; Xing, Junjie; Viollet, Benoit; Wang, Xian; Zhu, Yi; Zou, Ming-Hui

    2011-01-01

    Objective The aim of the present study was to determine whether mitochondrial uncoupling protein (UCP)-2 is required for AMPK-dependent angiogenesis in ischemia in vivo. Methods and Results Angiogenesis was assayed by monitoring endothelial tube formation (a surrogate for angiogenesis) in human umbilical vein endothelial cells (HUVECs), isolated mouse aortic endothelial cells (MAECs), and pulmomary microvascular endothelial cells (PMECs), or in ischemic thigh adductor muscles from wild-type (WT) mice or mice deficient in either AMPKα1 or AMPKα2. AMPK inhibition with pharmacological inhibitor (compound C) or genetic means (transfection of AMPKα-specific siRNA) significantly lowered the tube formation in HUVECs. Consistently, compared with WT mice, tube formation in MAECs isolated from either AMPKα1−/− or AMPKα2−/− mice, which exhibited oxidative stress and reduced expression of UCP2, were significantly impaired. In addition, adenoviral overexpression of UCP2, but not adenoviruses encoding green florescence protein (GFP), normalized tube formation in MAECs from either AMPKα1−/− or AMPKα2−/− mice. Similarly, supplementation with sodium nitroprusside (SNP), a nitric oxide (NO) donor, restored tube formation. Furthermore, ischemia significantly increased angiogenesis, serine 1177 phosphorylation of endothelial NO synthase (eNOS), and UCP2 in ischemic thigh adductor muscles from WT mice, but not from either AMPKα1−/− or AMPKα2−/− mice. Conclusion We conclude that AMPK-dependent UCP2 expression in endothelial cells promotes angiogenesis in vivo. PMID:21597006

  1. Studies on solid solutions based on layered honeycomb-ordered phases P2-Na{sub 2}M{sub 2}TeO{sub 6} (M=Co, Ni, Zn)

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

    Berthelot, Romain; Schmidt, Whitney; Sleight, A.W.

    2012-12-15

    Three complete solid solutions between the layered phases P2-Na{sub 2}M{sub 2}TeO{sub 6} (M=Co, Ni, Zn) have been prepared by conventional solid state method and investigated through X-ray diffraction, magnetism and optical measurements. All compositions are characterized by a M{sup 2+}/X{sup 6+} honeycomb ordering within the slabs and crystallize in a hexagonal unit cell. However, a structural transition based on a different stacking is observed as nickel (space group P6{sub 3}/mcm) is substituted by zinc or cobalt (space group P6{sub 3}22). All compositions exhibit a paramagnetic Curie-Weiss behavior at high temperatures; and the magnetic moment values confirm the presence of Ni{supmore » 2+} and/or Co{sup 2+} cations. The low-temperature antiferromagnetic order of Na{sub 2}Ni{sub 2}TeO{sub 6} and Na{sub 2}Co{sub 2}TeO{sub 6} is suppressed by zinc substitution. The color of the obtained compositions varies from pink, to light green and white when M=Co, Ni, Zn, respectively. - Graphical abstract: The comparison between the structure of Na{sub 2}Ni{sub 2}TeO{sub 6} (left) and Na{sub 2}M{sub 2}TeO{sub 6} (M=Co, Zn) (right) evidences the stacking difference with distinct atom sequences along the hexagonal c-axis. Highlights: Black-Right-Pointing-Pointer Solid solutions between lamellar phases Na{sub 2}M{sub 2}TeO{sub 6} (M=Co, Ni, Zn) are investigated. Black-Right-Pointing-Pointer A M{sup 2+}/X{sup 6+} honeycomb ordering characterized all the compositions. Black-Right-Pointing-Pointer A structural transition is shown when Ni is replaced by Co or Zn. Black-Right-Pointing-Pointer The low-temperature AFM ordering of Na{sub 2}Ni{sub 2}TeO{sub 6} and Na{sub 2}Co{sub 2}TeO{sub 6} is suppressed by zinc substitution. Black-Right-Pointing-Pointer Color changes from pink to light green and white when M=Co, Ni, Zn, respectively.« less

  2. Hydrothermal synthesis and structural characterization of an organic–inorganic hybrid sandwich-type tungstoantimonate [Cu(en){sub 2}(H{sub 2}O)]{sub 4}[Cu(en){sub 2}(H{sub 2}O){sub 2}][Cu{sub 2}Na{sub 4}(α-SbW{sub 9}O{sub 33}){sub 2}]·6H{sub 2}O

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

    Liu, Yingjie; College of Medicine, Henan University, Kaifeng, Henan 475004; Cao, Jing

    An organic–inorganic hybrid sandwich-type tungstoantimonate [Cu(en){sub 2}(H{sub 2}O)]{sub 4}[Cu(en){sub 2}(H{sub 2}O){sub 2}][Cu{sub 2}Na{sub 4}(α-SbW{sub 9}O{sub 33}){sub 2}]·6H{sub 2}O (1) has been synthesized by reaction of Sb{sub 2}O{sub 3}, Na{sub 2}WO{sub 4}·2H{sub 2}O, CuCl{sub 2}·2H{sub 2}O with en (en=ethanediamine) under hydrothermal conditions and structurally characterized by elemental analysis, inductively coupled plasma atomic emission spectrometry, IR spectrum and single-crystal X-ray diffraction. 1 displays a centric dimeric structure formed by two equivalent trivacant Keggin [α-SbW{sub 9}O{sub 33}]{sup 9−} subunits sandwiching a hexagonal (Cu{sub 2}Na{sub 4}) cluster. Moreover, those related hexagonal hexa-metal cluster sandwiched tungstoantimonates have been also summarized and compared. The variable-temperature magneticmore » measurements of 1 exhibit the weak ferromagnetic exchange interactions within the hexagonal (Cu{sub 2}Na{sub 4}) cluster mediated by the oxygen bridges. - Graphical abstract: An organic–inorganic hybrid (Cu{sub 2}Na{sub 4}) sandwiched tungstoantimonate [Cu(en){sub 2}(H{sub 2}O)]{sub 4}[Cu (en){sub 2}(H{sub 2}O){sub 2}][Cu{sub 2}Na{sub 4}(α-SbW{sub 9}O{sub 33}){sub 2}]·6H{sub 2}O was synthesized and magnetic properties was investigated. Display Omitted - Highlights: • Organic–inorganic hybrid sandwich-type tungstoantimonate. • (Cu{sub 2}Na{sub 4} sandwiched) tungstoantimonate [Cu{sub 2}Na{sub 4}(α-SbW{sub 9}O{sub 33}){sub 2}]{sup 10−}. • Ferromagnetic tungstoantimonate.« less

  3. A comparison of the interaction of nitric oxide with the heteropolytungstic acids H{sub 3}PW{sub 12}O{sub 40}, H{sub 0.5}Cs{sub 2.5}PW{sub 12}O{sub 40}, HMgPW{sub 12}O{sub 40}, H{sub 8}SiW{sub 11}O{sub 38}, H{sub 4}SiW{sub 12}O{sub 40}, and H{sub 10}CoW{sub 12}O{sub 42}

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

    Herring, A.M.; McCormick, R.L.; Boonrueng, S.R.

    2000-05-18

    The interaction between NO and the heteropolytungstic acids (HPAs) H{sub 3}PW{sub 12}O{sub 40} (HPW), H{sub 0.5}Cs{sub 2.5}PW{sub 12}O{sub 40} (HCsPW), HMgPW{sub 12}O{sub 40} (HMgPW), H{sub 8}SiW{sub 11}O{sub 38} (HSiW{sub 11}), H{sub 4}SiW{sub 12}O{sub 40} (HSiW), and H{sub 10}CoW{sub 12}{sub 42}(HCoW) in the presence of O{sub 2}(g) was investigated. The tools employed were in situ diffuse reflectance infrared spectroscopy, X-ray powder diffraction, and solid-state {sup 1}H NMR. It was determined that protons may either be present in the HPAs secondary structure as anhydrous protons or be bound to one or two water molecules as H{sub 3}O{sup +} or H{sub 5}O{sub 2}{supmore » +}, respectively. A previous investigation found that HPW sorbed NO into its bulk structure as NOH{sup +}, whereas the anhydrous potassium salt of HPW exhibited weak chemisorption of NO on its surface. In the present study, it was found that NO chemisorbed weakly on the surface of the anhydrous HCsPW. For HMgPW and HSiW, IR and NMR evidence suggests that water is present as H{sub 3}O{sup +}, and the formation of surface-bound NOH{sup +} was observed. Inclusion of NO into the secondary structure as NOH{sup +} was observed for HPW, HSiW{sub 11}, and HCoW. Literature data for HPW, as well as IR and NMR results reported here, indicate that these HPAs contain water as H{sub 5}O{sub 2}{sup +}. The presence of H{sub 5}O{sub 2}{sup +} is, therefore, a prerequisite for NO incorporation as NOH{sup +} in the HPA secondary structure. These HPAs exhibited two modes of NO bonding in the secondary structure: a strongly hydrogen-bound hydrated form, NOH{sup +}{center_dot}H{sub 2}O, and a more weakly bound anhydrous form, NOH{sup +}. Evidence suggests that it is the hydrated form that decomposes, yielding N{sub 2} upon rapid heating. Both NO-containing species interact with terminal and corner-sharing oxygen atoms of the Keggin ion. Anions that are held apart by terminal oxygen-hydrogen bonds have a larger

  4. Magnetotransport study of topological superconductor Cu<sub>0.10sub>Bi>2sub>Se>3sub> single crystal.

    PubMed

    Li, Mingtao; Fang, Yifei; Zhang, Jincang; Yi, Hemian; Zhou, Xingjiang; Lin, Chengtian

    2018-02-02

    We report a magnetotransport study of vortex-pinning in Cu<sub>0.10</sub>Bi<sub>2</sub>Se<sub>3</sub> single crystal. The sample is demonstrated to be in clean limit and absent of Pauli spin-limiting effect. Interestingly, the resistivity versus magnetic field shows an anomalously pronounced increase when approaching the superconducting-normal state boundary for both B∥ab and B∥c configurations. We have investigated the flux-flowing behavior under various magnetic field and temperatures, enabling us to establish its anisotropic vortex phase diagram. Our results suggest the Cu<sub>0.10</sub>Bi<sub>2</sub>Se<sub>3</sub> can be served as one unique material for exploring exotic surface vortex states in topological superconductors. © 2018 IOP Publishing Ltd.

  5. Phase equilibria in the NaF-CdO-NaPO{sub 3} system at 873 K and crystal structure and physico-chemical characterizations of the new Na{sub 2}CdPO{sub 4}F fluorophosphate

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

    Aboussatar, Mohamed; Laboratoire de Physico-Chimie de l’État Solide, Faculté des Sciences de Sfax, Université de Sfax, BP 1171, 3000 Sfax; Mbarek, Aïcha

    Isothermal sections of the diagram representing phase relationships in the NaF-CdO-NaPO{sub 3} system have been investigated by solid state reactions and powder X-ray diffraction. This phase diagram investigation confirms the polymorphism of the NaCdPO{sub 4} side component and the structure of the ß high temperature polymorph (orthorhombic, space group Pnma and unit cell parameters a=9.3118(2), b=7.0459(1), c=5.1849(1) Å has been refined. A new fluorophosphate, Na{sub 2}CdPO{sub 4}F, has been discovered and its crystal structure determined and refined from powder X-ray diffraction data. It exhibits a new 3D structure with orthorhombic symmetry, space group Pnma and unit cell parameters a=5.3731(1), b=6.8530(1),more » c=12.2691(2) Å. The structure is closely related to those of the high temperature polymorph of the nacaphite Na{sub 2}CaPO{sub 4}F and the fluorosilicate Ca{sub 2}NaSiO{sub 4}F but differs essentially in the cationic repartition since the structure is fully ordered with one Na site (8d) and one Cd site (4c). Relationships with other Na{sub 2}M{sup II}PO{sub 4}F (M{sup II}=Mg, Ca, Mn, Fe, Co, Ni) have been examined and the crystal-chemical and topographical analysis of these fluorophosphates is briefly reviewed. IR, Raman, optical and {sup 19}F, {sup 23}Na, {sup 31}P MAS NMR characterizations of Na{sub 2}CdPO{sub 4}F have been investigated. - Graphical abstract: The structure of the compound Na{sub 2}CdPO{sub 4}F, discovered during the study of the phase relationships in the NaF-CdO-NaPO{sub 3} system, has been determined and compared with other Na{sub 2}M{sup II}PO{sub 4}F fluorophosphates. - Highlights: • XRD analysis of the isothermal section of the NaF-CdO-NaPO{sub 3} system at 923 K. • Rietveld refinement of the high temperature polymorph β-NaCdPO{sub 4}. • Crystal structure of the new Na{sub 2}CdPO{sub 4}F fluorophosphate determined from powder XRD. • Crystal structure - composition relationships of Na{sub 2}M{sup II}PO{sub 4}F

  6. Cooperative heteroassembly of the adenoviral L4-22K and IVa2 proteins onto the viral packaging sequence DNA.

    PubMed

    Yang, Teng-Chieh; Maluf, Nasib Karl

    2012-02-21

    Human adenovirus (Ad) is an icosahedral, double-stranded DNA virus. Viral DNA packaging refers to the process whereby the viral genome becomes encapsulated by the viral particle. In Ad, activation of the DNA packaging reaction requires at least three viral components: the IVa2 and L4-22K proteins and a section of DNA within the viral genome, called the packaging sequence. Previous studies have shown that the IVa2 and L4-22K proteins specifically bind to conserved elements within the packaging sequence and that these interactions are absolutely required for the observation of DNA packaging. However, the equilibrium mechanism for assembly of IVa2 and L4-22K onto the packaging sequence has not been determined. Here we characterize the assembly of the IVa2 and L4-22K proteins onto truncated packaging sequence DNA by analytical sedimentation velocity and equilibrium methods. At limiting concentrations of L4-22K, we observe a species with two IVa2 monomers and one L4-22K monomer bound to the DNA. In this species, the L4-22K monomer is promoting positive cooperative interactions between the two bound IVa2 monomers. As L4-22K levels are increased, we observe a species with one IVa2 monomer and three L4-22K monomers bound to the DNA. To explain this result, we propose a model in which L4-22K self-assembly on the DNA competes with IVa2 for positive heterocooperative interactions, destabilizing binding of the second IVa2 monomer. Thus, we propose that L4-22K levels control the extent of cooperativity observed between adjacently bound IVa2 monomers. We have also determined the hydrodynamic properties of all observed stoichiometric species; we observe that species with three L4-22K monomers bound have more extended conformations than species with a single L4-22K bound. We suggest this might reflect a molecular switch that controls insertion of the viral DNA into the capsid.

  7. Protein kinase A-mediated cell proliferation in brown preadipocytes is independent of Erk1/2, PI{sub 3}K and mTOR

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

    Wang, Yanling; Sato, Masaaki; Guo, Yuan

    2014-10-15

    The physiological agonist norepinephrine promotes cell proliferation of brown preadipocytes during the process of tissue recruitment. In a primary culture system, cAMP mediates these adrenergic effects. In the present study, we demonstrated that, in contrast to other systems where the mitogenic effect of cAMP requires the synergistic action of (serum) growth factors, especially insulin/IGF, the cAMP effect in brown preadipocytes was independent of serum and insulin. Protein kinase A, rather than Epac, mediated the cAMP mitogenic effect. The Erk 1/2 family of MAPK, the PI{sub 3}K system and the mTOR complexes were all activated by cAMP, but these activations weremore » not necessary for cAMP-induced cell proliferation; a protein kinase C isoform may be involved in mediating cAMP-activated cell proliferation. We conclude that the generally acknowledged cellular mediators for induction of cell proliferation are not involved in this process in the brown preadipocyte system; this conclusion may be of relevance both for examination of mechanisms for induction of brown adipose tissue recruitment but also for understanding the mechanism behind e.g. certain endocrine neoplasias. - Highlights: • cAMP can mimick norepinephrine-induced proliferation of brown preadipocytes. • The cAMP-induced proliferation can occur in the absence of serum, of any other growth factors, and of insulin. • Erk1/2, PI{sub 3}K and mTOR are cAMP activated but not involved in induction of proliferation. • A Protein Kinase C member may be in the signalling cascade. • This pathway analysis may also be of importance for certain endocrine hyper- and neoplasias.« less

  8. Link between D sub 1 and D sub 2 dopamine receptors is reduced in schizophrenia and Huntington diseased brain

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

    Seeman, P.; Niznik, H.B.; Guan, H.C.

    1989-12-01

    Dopamine receptor types D{sub 1} and D{sub 2} can oppose enhance each other's actions for electrical, biochemical, and psychomotor effects. The authors report a D{sub 1}-D{sub 2} interaction in homogenized tissue as revealed by ligand binding. D{sub 2} agonists lowered the binding of ({sup 3}H)raclopride to D{sub 2} receptors in striatal and anterior pituitary tissues. Pretreating the tissue with the D{sub 1}-selective antagonist SCH 23390 prevented the agonist-induced decrease in ({sup 3}H)raclopride binding to D{sub 2} sites in the striatum but not in the anterior pituitary, which has no D{sub 1} receptors. Conversely, a dopamine-induced reduction in the binding ofmore » ({sup 3}H)SCH 23390 to D{sub 1} receptors could be prevented by the D{sub 2}-selective antagonist eticlopride. Receptor photolabeling experiments confirmed both these D{sub 1}-D{sub 2} interactions. The blocking effect by SCH 23390 was similar to that produced by a nonhydrolyzable guanine nucleotide analogue, and SCH 23390 reduced the number of agonist-labeled D{sub 2} receptors in the high-affinity state. Thus, the D{sub 1}-D{sub 2} link may be mediated by guanine nucleotide-binding protein components. The link may underlie D{sub 1}-D{sub 2} interactions influencing behavior, since the link was missing in over half the postmortem striata from patients with schizophrenia and Huntington disease (both diseases that show some hyperdopamine signs) but was present in human control, Alzheimer, and Parkinson striata.« less

  9. Hydrogen atom distribution and hydrogen induced site depopulation for the La{sub 2-x}Mg{sub x}Ni{sub 7}-H system

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

    Guzik, Matylda N., E-mail: Matylda.Guzik@ife.no; Physics Department, Institute for Energy Technology, P.O. Box 40, NO-2027 Kjeller; Hauback, Bjorn C.

    2012-02-15

    La{sub 2-x}Mg{sub x}Ni{sub 7} and its hydrides/deuterides were investigated by high resolution synchrotron powder X-ray and neutron diffraction. Upon deuteration the single phase sample of the intermetallic compound with the refined composition La{sub 1.63}Mg{sub 0.37}Ni{sub 7} (space group: P6{sub 3}/mmc) expands isotropically, in contrast to the Mg free phase. The hydrogen uptake, {approx}9 D/f.u., is higher than in La{sub 2}Ni{sub 7}D{sub 6.5}. The refined composition accounts for La{sub 1.63}Mg{sub 0.37}Ni{sub 7}D{sub 8.8} (beta-phase). Rietveld refinements using the neutron and synchrotron diffraction data suggest that deuterium atoms occupy 5 different interstitial sites within both AB{sub 2} and AB{sub 5} slabs, eithermore » in an ordered or a disordered way. All determined D sites have an occupancy >50% and the shortest D-D contact is 1.96(3) A. It is supposed that a competition between the tendency to form directional bonds and repulsive D-D (H-H) interactions is the most important factor that influences the distribution of deuterium atoms in this structure. A hitherto unknown second, alpha-phase with composition La{sub 1.63}Mg{sub 0.37}Ni{sub 7}D{sub 0.56}, crystallizing with the same hexagonal symmetry as La{sub 1.63}Mg{sub 0.37}Ni{sub 7}D{sub 8.8}, has been discovered. The unit cell parameters for this D-poor phase differ slightly from those of the intermetallic. Alpha-phase displays only one D site (4f, space group: P6{sub 3}/mmc) occupied >50%, which is not populated in the D-rich beta-phase. This hydrogen/deuterium induced site depopulation can be explained by repulsive D-D (H-H) interactions that are likely to influence non-occupancy of certain interstices in metal lattice when absorbing hydrogen. - Graphical abstract: The detailed D atoms arrangement in La{sub 1.63}Mg{sub 0.37}Ni{sub 7}D{sub 8.8} differs significantly from the previously reported La{sub 1.5}Mg{sub 0.5}Ni{sub 7}D{sub 8.9(9.1)}. The present model consists of only five deuterium

  10. Conformational Sub-states and Populations in Enzyme Catalysis

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

    Agarwal, Pratul K; Doucet, Nicholas; Chennubholta, C

    reactants in the active site, chemical turnover, and release of products. In addition to formation of crucial structural interactions between enzyme and substrate(s), coordinated motions within the enzyme substrate complex allow reaction to proceed at a much faster rate, compared to the reaction in solution and in the absence of enzyme. An increasing number of enzyme systems show the presence of conserved protein motions that are important for function. A wide variety of motions are naturally sampled (over femtosecond to millisecond time-scales) as the enzyme complex moves along the energetic landscape, driven by temperature and dynamical events from the surroundingmore » environment. Areas of low energy along the landscape form conformational sub-states, which show higher conformational populations than surrounding areas. A small number of these protein conformational sub-states contain functionally important structural and dynamical features, which assist the enzyme mechanism along the catalytic cycle. Identification and characterization of these higher-energy (also called excited) sub-states and the associated populations are challenging, as these sub-states are very short-lived and therefore rarely populated. Specialized techniques based on computer simulations, theoretical modeling, and nuclear magnetic resonance have been developed for quantitative characterization of these sub-states and populations. This chapter discusses these techniques and provides examples of their applications to enzyme systems.« less

  11. Understanding the molecular basis of plant growth promotional effect of Pseudomonas fluorescens on rice through protein profiling.

    PubMed

    Kandasamy, Saveetha; Loganathan, Karthiba; Muthuraj, Raveendran; Duraisamy, Saravanakumar; Seetharaman, Suresh; Thiruvengadam, Raguchander; Ponnusamy, Balasubramanian; Ramasamy, Samiyappan

    2009-12-24

    Plant Growth Promoting Rhizobacteria (PGPR), Pseudomonas fluorescens strain KH-1 was found to exhibit plant growth promotional activity in rice under both in-vitro and in-vivo conditions. But the mechanism underlying such promotional activity of P. fluorescens is not yet understood clearly. In this study, efforts were made to elucidate the molecular responses of rice plants to P. fluorescens treatment through protein profiling. Two-dimensional polyacrylamide gel electrophoresis strategy was adopted to identify the PGPR responsive proteins and the differentially expressed proteins were analyzed by mass spectrometry. Priming of P. fluorescens, 23 different proteins found to be differentially expressed in rice leaf sheaths and MS analysis revealed the differential expression of some important proteins namely putative p23 co-chaperone, Thioredoxin h- rice, Ribulose-bisphosphate carboxylase large chain precursor, Nucleotide diPhosphate kinase, Proteosome sub unit protein and putative glutathione S-transferase protein. Functional analyses of the differential proteins were reported to be directly or indirectly involved in growth promotion in plants. Thus, this study confirms the primary role of PGPR strain KH-1 in rice plant growth promotion.

  12. Sub-cellular localisation studies may spuriously detect the Yes-associated protein, YAP, in nucleoli leading to potentially invalid conclusions of its function.

    PubMed

    Finch, Megan L; Passman, Adam M; Strauss, Robyn P; Yeoh, George C; Callus, Bernard A

    2015-01-01

    The Yes-associated protein (YAP) is a potent transcriptional co-activator that functions as a nuclear effector of the Hippo signaling pathway. YAP is oncogenic and its activity is linked to its cellular abundance and nuclear localisation. Activation of the Hippo pathway restricts YAP nuclear entry via its phosphorylation by Lats kinases and consequent cytoplasmic retention bound to 14-3-3 proteins. We examined YAP expression in liver progenitor cells (LPCs) and surprisingly found that transformed LPCs did not show an increase in YAP abundance compared to the non-transformed LPCs from which they were derived. We then sought to ascertain whether nuclear YAP was more abundant in transformed LPCs. We used an antibody that we confirmed was specific for YAP by immunoblotting to determine YAP's sub-cellular localisation by immunofluorescence. This antibody showed diffuse staining for YAP within the cytosol and nuclei, but, noticeably, it showed intense staining of the nucleoli of LPCs. This staining was non-specific, as shRNA treatment of cells abolished YAP expression to undetectable levels by Western blot yet the nucleolar staining remained. Similar spurious YAP nucleolar staining was also seen in mouse embryonic fibroblasts and mouse liver tissue, indicating that this antibody is unsuitable for immunological applications to determine YAP sub-cellular localisation in mouse cells or tissues. Interestingly nucleolar staining was not evident in D645 cells suggesting the antibody may be suitable for use in human cells. Given the large body of published work on YAP in recent years, many of which utilise this antibody, this study raises concerns regarding its use for determining sub-cellular localisation. From a broader perspective, it serves as a timely reminder of the need to perform appropriate controls to ensure the validity of published data.

  13. Sub-Cellular Localisation Studies May Spuriously Detect the Yes-Associated Protein, YAP, in Nucleoli Leading to Potentially Invalid Conclusions of Its Function

    PubMed Central

    Finch, Megan L.; Passman, Adam M.; Strauss, Robyn P.; Yeoh, George C.; Callus, Bernard A.

    2015-01-01

    The Yes-associated protein (YAP) is a potent transcriptional co-activator that functions as a nuclear effector of the Hippo signaling pathway. YAP is oncogenic and its activity is linked to its cellular abundance and nuclear localisation. Activation of the Hippo pathway restricts YAP nuclear entry via its phosphorylation by Lats kinases and consequent cytoplasmic retention bound to 14-3-3 proteins. We examined YAP expression in liver progenitor cells (LPCs) and surprisingly found that transformed LPCs did not show an increase in YAP abundance compared to the non-transformed LPCs from which they were derived. We then sought to ascertain whether nuclear YAP was more abundant in transformed LPCs. We used an antibody that we confirmed was specific for YAP by immunoblotting to determine YAP’s sub-cellular localisation by immunofluorescence. This antibody showed diffuse staining for YAP within the cytosol and nuclei, but, noticeably, it showed intense staining of the nucleoli of LPCs. This staining was non-specific, as shRNA treatment of cells abolished YAP expression to undetectable levels by Western blot yet the nucleolar staining remained. Similar spurious YAP nucleolar staining was also seen in mouse embryonic fibroblasts and mouse liver tissue, indicating that this antibody is unsuitable for immunological applications to determine YAP sub-cellular localisation in mouse cells or tissues. Interestingly nucleolar staining was not evident in D645 cells suggesting the antibody may be suitable for use in human cells. Given the large body of published work on YAP in recent years, many of which utilise this antibody, this study raises concerns regarding its use for determining sub-cellular localisation. From a broader perspective, it serves as a timely reminder of the need to perform appropriate controls to ensure the validity of published data. PMID:25658431

  14. Electron density distribution and disordered crystal structure of 15R-SiAlON, SiAl{sub 4}O{sub 2}N{sub 4}

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

    Banno, Hiroki; Hanai, Takaaki; Asaka, Toru

    2014-03-15

    The crystal structure of SiAl{sub 4}O{sub 2}N{sub 4} was characterized by laboratory X-ray powder diffraction (CuKα{sub 1}). The title compound is trigonal with space group R3-bar m. The hexagonal unit-cell dimensions (Z=3) are a=0.301332(3) nm, c=4.18616(4) nm and V=0.3291825(5) nm{sup 3}. The initial structural model was successfully derived by the charge-flipping method and further refined by the Rietveld method. The final structural model showed the positional disordering of one of the three (Si,Al) sites. The maximum-entropy method-based pattern fitting (MPF) method was used to confirm the validity of the split-atom model, in which conventional structure bias caused by assuming intensitymore » partitioning was minimized. The reliability indices calculated from the MPF were R{sub wp}=5.05%, S (=R{sub wp}/R{sub e})=1.21, R{sub p}=3.77%, R{sub B}=1.29% and R{sub F}=1.01%. The disordered crystal structure was successfully described by overlapping three types of domains with ordered atom arrangements. The distribution of atomic positions in one of the three types of domains can be achieved in the space group R3-bar m. The atom arrangements in the other two types of domains are noncentrosymmetrical with the space group R3m. These two structural configurations are related by the pseudo-symmetry inversion. -- Graphical abstract: A bird's eye view of electron densities up to 75.3% (0.133 nm{sup −3}) of the maximum on the plane parallel to (110) with the corresponding atomic arrangements of SiAl{sub 4}O{sub 2}N{sub 4}. Highlights: • Crystal structure of SiAl{sub 4}O{sub 2}N{sub 4} is determined by laboratory X-ray powder diffraction. • The atom arrangements are represented by the split-atom model. • The maximum-entropy method-based pattern fitting method is used to confirm the validity of the model. • The disordered structure is described by overlapping three types of domains with ordered atom arrangements.« less

  15. Five inorganic–organic hybrids based on Keggin polyanion [SiMo{sub 12}O{sub 40}]{sup 4−}: From 0D to 2D network

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

    Yu, Xiao-Yang, E-mail: yangyangyu0103@sohu.com; Jilin Institute of Chemical Technology, Jilin City, Jilin 132022; Cui, Xiao-Bing

    2014-01-15

    Five new inorganic–organic hybrids based on 4,4′-bipyridine and Keggin-type polyoxometalate [SiMo{sub 12}O{sub 40}]{sup 4−}, (SiMo{sub 12}O{sub 40})(H{sub 2}bipy){sub 2}·2H{sub 2}O (1), [Cu(Hbipy){sub 4}(HSiMo{sub 12}O{sub 40})(SiMo{sub 12}O{sub 40})](H{sub 2}bipy){sub 0.5}·7H{sub 2}O (2), [Cu{sub 2}(Hbipy){sub 6}(bipy)(SiMo{sub 12}O{sub 40}){sub 3}](Hbipy){sub 2}·6H{sub 2}O (3), [Cu(bipy){sub 2}(SiMo{sub 12}O{sub 40})](H{sub 2}bipy)·2H{sub 2}O (4) and [Cu{sub 2}(bipy){sub 4}(H{sub 2}O){sub 4}](SiMo{sub 12}O{sub 40})·13H{sub 2}O (5) (bipy=4,4′-bipyridine), have been hydrothermally synthesized. 1 consists of H{sub 2}bipy{sup 2+} and [SiMo{sub 12}O{sub 40}]{sup 4−} units. In 2, two [SiMo{sub 12}O{sub 40}]{sup 4−} are bridged by [Cu(Hbipy){sub 4}]{sup 6+} to form a [Cu(Hbipy){sub 4}(SiMo{sub 12}O{sub 40}){sub 2}]{sup 2−} dimmer. In 3, [SiMo{submore » 12}O{sub 40}]{sup 4−} polyanions acting as bidentated bridging ligands and monodentated auxiliary ligands connect [Cu{sub 2}(Hbipy){sub 6}(bipy)]{sup 8+} units into a 1D zigzag chain. In 4, [SiMo{sub 12}O{sub 40}]{sup 4−} polyanions bridge neighboring 1D [Cu(bipy){sub 2}]{sup 2+} double chains into a 2D extended layer. In 5, [SiMo{sub 12}O{sub 40}]{sup 4−} polyanions acting as templates site alternately upon the grids from both sides of the square grid [Cu{sub 2}(bipy){sub 4}(H{sub 2}O){sub 4}]{sup 4+} layer. In addition, the electrochemical behaviors of 1, 3 and 4 and the photocatalysis property of 1 have been investigated. - Graphical abstract: Five new compounds based on [SiMo{sub 12}O{sub 40}]{sup 4−} have been successfully generated. [SiMo{sub 12}O{sub 40}]{sup 4−} anions play different roles in the structures of the five compounds. Display Omitted - Highlights: • Five new compounds based on [SiMo{sub 12}O{sub 40}]{sup 4−} have been generated. • [SiMo{sub 12}O{sub 40}]{sup 4−} anions play different roles in the five structures. • The electrochemical behaviors of 1, 3 and 4

  16. Pre-fractionation strategies to resolve pea (Pisum sativum) sub-proteomes

    PubMed Central

    Meisrimler, Claudia-Nicole; Menckhoff, Ljiljana; Kukavica, Biljana M.; Lüthje, Sabine

    2015-01-01

    Legumes are important crop plants and pea (Pisum sativum L.) has been investigated as a model with respect to several physiological aspects. The sequencing of the pea genome has not been completed. Therefore, proteomic approaches are currently limited. Nevertheless, the increasing numbers of available EST-databases as well as the high homology of the pea and medicago genome (Medicago truncatula Gaertner) allow the successful identification of proteins. Due to the un-sequenced pea genome, pre-fractionation approaches have been used in pea proteomic surveys in the past. Aside from a number of selective proteome studies on crude extracts and the chloroplast, few studies have targeted other components such as the pea secretome, an important sub-proteome of interest due to its role in abiotic and biotic stress processes. The secretome itself can be further divided into different sub-proteomes (plasma membrane, apoplast, cell wall proteins). Cell fractionation in combination with different gel-electrophoresis, chromatography methods and protein identification by mass spectrometry are important partners to gain insight into pea sub-proteomes, post-translational modifications and protein functions. Overall, pea proteomics needs to link numerous existing physiological and biochemical data to gain further insight into adaptation processes, which play important roles in field applications. Future developments and directions in pea proteomics are discussed. PMID:26539198

  17. Structural, optical, and magnetic properties of Na{sub 8}Eu{sub 2}(Si{sub 2}S{sub 6}){sub 2} and Na{sub 8}Eu{sub 2}(Ge{sub 2}S{sub 6}){sub 2}: Europium(II) quaternary chalcogenides that contain an ethane-like (Si{sub 2}S{sub 6}){sup 6−} or (Ge{sub 2}S{sub 6}){sup 6−} moiety

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

    Choudhury, Amitava, E-mail: choudhurya@mst.edu; Ghosh, Kartik; Grandjean, Fernande

    2015-03-15

    Two isostructural europium(II) quaternary chalcogenides, Na{sub 8}Eu{sub 2}(Si{sub 2}S{sub 6}){sub 2}, 1, and Na{sub 8}Eu{sub 2}(Ge{sub 2}S{sub 6}){sub 2}, 2, containing an ethane-like (Si{sub 2}S{sub 6}){sup 6−} or (Ge{sub 2}S{sub 6}){sup 6−} moiety have been synthesized by employing the polychalcogenide molten flux method. Single-crystal X-ray diffraction reveals that both compounds crystallize in the C2/m space group, and their structures contain layers of ([Na{sub 2}Eu{sub 2}(Si{sub 2}S{sub 6}){sub 2}]{sup 6−}){sub ∞} or ([Na{sub 2}Eu{sub 2}(Ge{sub 2}S{sub 6}){sub 2}]{sup 6−}){sub ∞} anions held together by six interlayer sodium cations to yield (Na{sub 6}[Na{sub 2}Eu{sub 2}(Si{sub 2}S{sub 6}){sub 2}]){sub ∞} and (Na{sub 6}[Na{submore » 2}Eu{sub 2}(Ge{sub 2}S{sub 6}){sub 2}]){sub ∞}. Compound 2 is a semiconductor with an optical band gap of 2.15(2) eV. The temperature dependence of the magnetic susceptibility indicates that compounds 1 and 2 are paramagnetic with μ{sub eff}=7.794(1) μ{sub B} per Eu and g=1.964(1) for 1 and μ{sub eff}=8.016(1) μ{sub B} per Eu and g=2.020(1) for 2, moments that are in good agreement with the europium(II) spin-only moment of 7.94 μ{sub B}. The europium-151 Mössbauer isomer shift of 2 confirms the presence of europium(II) cations with an electronic configuration between [Xe]4f{sup 6.81} and 4f{sup 7}6s{sup 0.32}. - Graphical abstract: TOC figure caption: structure of Na{sub 8}Eu{sub 2}(Si{sub 2}S{sub 6}){sub 2} viewed along the a-axis showing the filling of A–B and B–A types of anion layers with two different types of cations. - Highlights: • Synthesis of quaternary europium chalcogenides containing ethane-like dimer. • Structural characterization employing single-crystal X-ray diffraction. • Mössbauer spectroscopy and magnetic measurements confirm presence of Eu(II)« less

  18. Adenoviral Gene Transfer of Hepatic Stimulator Substance Confers Resistance Against Hepatic Ischemia–Reperfusion Injury by Improving Mitochondrial Function

    PubMed Central

    Jiang, Shu-Jun; Li, Wen

    2013-01-01

    Abstract Hepatic stimulator substance (HSS) has been suggested to protect liver cells from various toxins. However, the precise role of HSS in hepatic ischemia–reperfusion (I/R) injury remains unknown. This study aims to elucidate whether overexpression of HSS could attenuate hepatic ischemia–reperfusion injury and its possible mechanisms. Both in vivo hepatic I/R injury in mice and in vitro hypoxia–reoxygenation (H/R) in a cell model were used to evaluate the effect of HSS protection after adenoviral gene transfer. Moreover, a possible mitochondrial mechanism of HSS protection was investigated. Efficient transfer of the HSS gene into liver inhibited hepatic I/R injury in mice, as evidenced by improvement in liver function tests, the preservation of hepatic morphology, and a reduction in hepatocyte apoptosis. HSS overexpression also inhibited H/R-induced cell death, as detected by cell viability and cell apoptosis assays. The underlying mechanism of this hepatic protection might involve the attenuation of mitochondrial dysfunction and mitochondrial-dependent cell apoptosis, as shown by the good preservation of mitochondrial ultrastructure, mitochondrial membrane potential, and the inhibition of cytochrome c leakage and caspase activity. Moreover, the suppression of H/R-induced mitochondrial ROS production and the maintenance of mitochondrial respiratory chain complex activities may participate in this mechanism. This new function of HSS expands the possibility of its application for the prevention of I/R injury, such as hepatic resection and liver transplantation in clinical practice. PMID:23461564

  19. Ce{sub 11}Ge{sub 3.73(2)}In{sub 6.27}: Solid-state synthesis, crystal structure and site-preference

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

    Jeon, Beom-Yong; Nam, Gnu; Lee, Dong Woo

    A novel intermetallic compound of Ce{sub 11}Ge{sub 3.73(2)}In{sub 6.27} has been synthesized through the high-temperature solid-state reaction using Nb-ampoules. A batch of well grown block-/short bar-shaped single-crystals has been obtained, and the crystal structure of the title compound has been characterized by single-crystal X-ray diffraction analyses. Ce{sub 11}Ge{sub 3.73(2)}In{sub 6.27} adopts the Ho{sub 11}Ge{sub 10}-type structure belonging to the tetragonal space group I4/mmm (Z=4, Pearson symbol tI84) with nine crystallographically unique atomic positions in the asymmetric unit. The lattice parameters are a=12.0163(1) Å and c=16.5396(2) Å. The overall crystal structure can simply be depicted as an assembly of three differentmore » types of co-facial cationic polyhedra centered by anions, which is further enclosed by the three-dimensional (3-D) cage-like anionic framework. The extra amount of In is observed in one of three isolated anionic sites resulting in introducing the Ge/In-mixed site at the Wyckoff 4e site. This unique site-preference of In substitution for Ge at the 4e site has been enlightened via the atomic size-aspect which was fully supported and rationalized by the site- and bond-energies analyses using tight-binding linear muffin-tin orbital (TB-LMTO) calculations. Energy-dispersive X-ray spectroscopy (EDS), density of states (DOS), crystal orbital Hamilton population (COHP), and electron localization function (ELF) analyses for the title compound are also presented. Magnetic susceptibility measurement proves that an antiferromagnetic ordering of Ce atoms at a low temperature with a paramagnetic Curie temperature of −23.2 K. - Graphical abstract: Reported is experimental and theoretical investigations for Ce{sub 11}Ge{sub 3.73(2)}In{sub 6.27}, which is the first reported example having the extra amounts of In substitution for Ge at one of three “isolated” anionic sites in the Ho{sub 11}Ge{sub 10}-type phase. The observed In site

  20. Age-related pathology after adenoviral overexpression of the leucine-rich repeat kinase 2 in the mouse striatum.

    PubMed

    Kritzinger, Astrid; Ferger, Boris; Gillardon, Frank; Stierstorfer, Birgit; Birk, Gerald; Kochanek, Stefan; Ciossek, Thomas

    2018-06-01

    Mutations in leucine-rich repeat kinase 2 (LRRK2) age-dependently cause Parkinson's disease and are associated with several inflammatory diseases. So far, the potential role of LRRK2 expression in glial cells as mediators of neuroinflammation and the influence of aging have not been investigated in viral vector-based LRRK2 animal models. In this study, we compared the effect of striatal injection of high-capacity adenoviral vectors expressing either a kinase-overactive LRRK2 with the familial G2019S mutation or a kinase-inactive LRRK2 variant in young and old C57BL/6J mice. The intrinsic adenovirus tropism guided preferentially glial transduction, and the vector design led to stable expression for at least 6 months. In histopathological analysis, young mice expressing either LRRK2 variant presented with transient vacuolization of striatal white fiber tracts accompanied by accumulation of microglial cells and astrogliosis, but inflammation resolved without permanent damage. Old mice had a stronger and prolonged inflammatory reaction and experienced permanent damage in form of partial neuron loss after 3 months exclusively in case of LRRK2_G2019S expression. The autophagic receptor p62 accumulated in cells with high levels of either LRRK2 variant, even more so in old mice. We conclude that the aging mouse brain is more susceptible to LRRK2-associated pathology, and in this model, glial LRRK2 expression significantly contributed to neuroinflammation, ultimately causing neurodegeneration. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.