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Sample records for neuraminidase na genes

  1. Cloning of neuraminidase (NA) gene and identification of its antiviral ...

    African Journals Online (AJOL)

    ... the sialic acid receptor required by virus infection of the host cell surface which protects the host from virus damage. In order to explore a new idea to use neuraminidase (NA) gene and produce disease-resistant transgenic poultry, prokaryotic expression vector pGEX-NA was constructed to make NA polyclone antibody.

  2. Cloning of neuraminidase (NA) gene and identification of its antiviral ...

    African Journals Online (AJOL)

    user

    2012-06-12

    Jun 12, 2012 ... neuraminidase (NA) gene and produce disease-resistant transgenic poultry, prokaryotic expression ... transfected cells were challenged by Newcastle disease virus (NDV), the morphology of CEF cells were observed to detect the .... eukaryotic expression vector pcDNA3.0-NA and pcDNA3.0/EGFP-. NA.

  3. Partial antiviral activities detection of chicken Mx jointing with neuraminidase gene (NA against Newcastle disease virus.

    Directory of Open Access Journals (Sweden)

    Yani Zhang

    Full Text Available As an attempt to increase the resistance to Newcastle Disease Virus (NDV and so further reduction of its risk on the poultry industry. This work aimed to build the eukaryotic gene co-expression plasmid of neuraminidase (NA gene and myxo-virus resistance (Mx and detect the gene expression in transfected mouse fibroblasts (NIH-3T3 cells, it is most important to investigate the influence of the recombinant plasmid on the chicken embryonic fibroblasts (CEF cells. cDNA fragment of NA and mutant Mx gene were derived from pcDNA3.0-NA and pcDNA3.0-Mx plasmid via PCR, respectively, then NA and Mx cDNA fragment were inserted into the multiple cloning sites of pVITRO2 to generate the eukaryotic co-expression plasmid pVITRO2-Mx-NA. The recombinant plasmid was confirmed by restriction endonuclease treatment and sequencing, and it was transfected into the mouse fibroblasts (NIH-3T3 cells. The expression of genes in pVITRO2-Mx-NA were measured by RT-PCR and indirect immunofluorescence assay (IFA. The recombinant plasmid was transfected into CEF cells then RT-PCR and the micro-cell inhibition tests were used to test the antiviral activity for NDV. Our results showed that co-expression vector pVITRO2-Mx-NA was constructed successfully; the expression of Mx and NA could be detected in both NIH-3T3 and CEF cells. The recombinant proteins of Mx and NA protect CEF cells from NDV infection until after 72 h of incubation but the individually mutagenic Mx protein or NA protein protects CEF cells from NDV infection till 48 h post-infection, and co-transfection group decreased significantly NDV infection compared with single-gene transfection group (P<0. 05, indicating that Mx-NA jointing contributed to delaying the infection of NDV in single-cell level and the co-transfection of the jointed genes was more powerful than single one due to their synergistic effects.

  4. Baculovirus Surface Display Using Infuenza Neuraminidase (NA Transmembrane Anchor

    Directory of Open Access Journals (Sweden)

    Irisa Trianti

    2016-11-01

    Full Text Available Baculovirus surface display has been employed as an excellent tools for presentation of foreign peptides and proteins on virus surface with native conformation, functions and immunogenicity. A baculovirus major envelope protein, gp64, or a capsid protein, vp39 are generally used as fusion partners for displaying of polypeptides on the surface of virions. Alternatively, a membrane anchoring domain of vesicular stomatitis virus G protein (VSV-G can also be used. In this study, an influenza neuraminidase (NA was proposed as a new membrane anchor for the display of Angiotensin II (AngII, DRVYIHPFHL, peptides. The AngII peptides were inserted into NA by replacing NA amino acid number 60-67 with AngII, and then integrated into a baculovirus genome. A recombinant baculovirus expressing the NA fusion-AngII peptides was generated from infected insect cells. Those peptides were found to express and translocated on the membrane of the baculovirus infected insect cell (Sf9 cell as detected by immunocytochemistry using anti-AngII monoclonal antibody. Upon budding of the recombinant baculovirus progenies through the insect cells membrane, the recombinant NA-AngII peptides was acquired to envelopes of the new baculovirus progenies. The conformation of NA on baculovirus surface was not affected by the deletion, as the 55 kDa band of NA can be detected from Western Blotting analysis by specific anti-NA monoclonal antibody. In addition, the same protein was also found by anti-AngII antibody indicating that the AngII peptides had been successfully fused with the recombinant NA. Interestingly, electron microscopy analysis demonstrated that not only the recombinant baculovirus displaying AngII peptides were generated by infected insect cells, but also the NA virus-like-particle displaying AngII peptides.

  5. Sialidosis and galactosialidosis: chromosomal assignment of two genes associated with neuraminidase-deficiency disorders

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    Mueller, O.T.; Henry, W.M.; Haley, L.L.; Byers, M.G.; Eddy, R.L.; Shows, T.B.

    1986-03-01

    The inherited human disorders sialidosis and galactosialidosis are the result of deficiencies of glycoprotein-specific ..cap alpha..-neuraminidase (acylneuraminyl hydrolase, EC 3.2.1.18; sialidase) activity. Two genes were determined to be necessary for expression of neuraminidase by using human-mouse somatic cell hybrids segregating human chromosomes. A panel of mouse RAG-human hybrid cells demonstrated a single-gene requirement for human neuraminidase and allowed assignment of this gene to the (pter ..-->.. q23) region of chromosome 10. A second panel of mouse thymidine kinase (TK)-deficient LM/TK/sup -/-human hybrid cells demonstrated that human neuraminidase activity required both chromosomes 10 and 20 to be present. Analysis of human neuraminidase expression in interspecific hybrid cells or polykaryocytes formed from fusion of mouse RAG or LM/TK/sup -/ cell lines with human sialidosis or galactosialidosis fibroblasts indicated that the RAG cell line complemented the galactosialidosis defect, but the LM/TK/sup -/ cell line did not. This eliminates the requirement for this gene in RAG-human hybrid cells and explains the different chromosome requirements of these two hybrid panels. Fusion of LM/TK/sup -/ cell hybrids lacking chromosome 10 or 20 and neuraminidase-deficient fibroblasts confirmed by complementation analysis that the sialidosis disorder results from a mutation on chromosome 10, presumably encoding the neuraminidase structural gene. Galactosialidosis is caused by a mutation in a second gene required for neuraminidase expression located on chromosome 20.

  6. Sialidosis and galactosialidosis: chromosomal assignment of two genes associated with neuraminidase-deficiency disorders.

    Science.gov (United States)

    Mueller, O T; Henry, W M; Haley, L L; Byers, M G; Eddy, R L; Shows, T B

    1986-01-01

    The inherited human disorders sialidosis and galactosialidosis are the result of deficiencies of glycoprotein-specific alpha-neuraminidase (acylneuraminyl hydrolase, EC 3.2.1.18; sialidase) activity. Two genes were determined to be necessary for expression of neuraminidase by using human-mouse somatic cell hybrids segregating human chromosomes. A panel of mouse RAG-human hybrid cells demonstrated a single-gene requirement for human neuraminidase and allowed assignment of this gene to the (pter----q23) region of chromosome 10. A second panel of mouse thymidine kinase (TK)-deficient LM/TK- -human hybrid cells demonstrated that human neuraminidase activity required both chromosomes 10 and 20 to be present. Analysis of human neuraminidase expression in interspecific hybrid cells or polykaryocytes formed from fusion of mouse RAG (hypoxanthine/guanine phosphoribosyltransferase deficient) or LM/TK- cell lines with human sialidosis or galactosialidosis fibroblasts indicated that the RAG cell line complemented the galactosialidosis defect, but the LM/TK- cell line did not. This eliminates the requirement for this gene in RAG-human hybrid cells and explains the different chromosome requirements of these two hybrid panels. Fusion of LM/TK- cell hybrids lacking chromosome 10 or 20 (phenotype 10+,20- and 10-,20+) and neuraminidase-deficient fibroblasts confirmed by complementation analysis that the sialidosis disorder results from a mutation on chromosome 10, presumably encoding the neuraminidase structural gene. Galactosialidosis is caused by a mutation in a second gene required for neuraminidase expression located on chromosome 20. PMID:3081902

  7. Sialidosis Type 1 with a Novel Mutation in the Neuraminidase-1 (NEU1) Gene.

    Science.gov (United States)

    Gowda, Vykuntaraju K; Srinivasan, Varun M; Benakappa, Naveen; Benakappa, Asha

    2017-05-01

    A patient with Sialidosis type 1 with a novel variation in neuraminidase-1 (NEU1) is described. The patient developed ataxia and myoclonus at 9 y of age. He was born to a second degree consanguineous marriage couple. On examination child had cerebellar signs and bilateral macular cherry-red spots. MRI of the brain and electroencephalogram were normal. The enzyme analysis revealed deficiency of neuraminidase. Genetic analysis identified novel homozygous missense mutation c.742G > T (p.G248C) in exon 4 of NEU1 gene. At 13 y of age, the ataxia and had myoclonus progressed.

  8. A broad spectrum, one-step reverse-transcription PCR amplification of the neuraminidase gene from multiple subtypes of influenza A virus

    Directory of Open Access Journals (Sweden)

    Chen Wenbin

    2008-07-01

    Full Text Available Abstract Background The emergence of high pathogenicity strains of Influenza A virus in a variety of human and animal hosts, with wide geographic distribution, has highlighted the importance of rapid identification and subtyping of the virus for outbreak management and treatment. Type A virus can be classified into subtypes according to the viral envelope glycoproteins, hemagglutinin and neuraminidase. Here we review the existing specificity and amplification of published primers to subtype neuraminidase genes and describe a new broad spectrum primer pair that can detect all 9 neuraminidase subtypes. Results Bioinformatic analysis of 3,337 full-length influenza A neuraminidase segments in the NCBI database revealed semi-conserved regions not previously targeted by primers. Two degenerate primers with M13 tags, NA8F-M13 and NA10R-M13 were designed from these regions and used to generate a 253 bp cDNA product. One-step RT-PCR testing was successful in 31/32 (97% cases using a touchdown protocol with RNA from over 32 different cultured influenza A virus strains representing the 9 neuraminidase subtypes. Frozen blinded clinical nasopharyngeal aspirates were also assayed and were mostly of subtype N2. The region amplified was direct sequenced and then used in database searches to confirm the identity of the template RNA. The RT-PCR fragment generated includes one of the mutation sites related to oseltamivir resistance, H274Y. Conclusion Our one-step RT-PCR assay followed by sequencing is a rapid, accurate, and specific method for detection and subtyping of different neuraminidase subtypes from a range of host species and from different geographical locations.

  9. Clinical presentation of congenital sialidosis in a patient with a neuraminidase gene frameshift mutation.

    Science.gov (United States)

    Buchholz, T; Molitor, G; Lukong, K E; Praun, M; Genzel-Boroviczény, O; Freund, M; Pshezhetsky, A V; Schulze, A

    2001-01-01

    Congenital sialidosis is a rare lysosomal storage disease caused by a primary neuraminidase deficiency which results from defects in the neuraminidase gene on chromosome 6p. The inheritance is autosomal recessive. Patients exhibit excessive urinary excretion of bound sialic acid and decreased or undetectable amounts of neuraminidase activity in various tissues. The clinical expression is variable, but ascites and hepatosplenomegaly are hallmarks of the disease. Skeletal abnormalities, facial dysmorphism and inguinal herniae have been described in most of the few reported cases. We describe a baby girl with biochemically proven sialidosis, who in addition to the above clinical features, had severely dilated coronary arteries, excessive retinal vascular tortuosity and an erythematous, macular rash. Homozygosity for a frameshift mutation at residue 623 of the neuraminidase cDNA was found. We speculate that the additional features found in our patient might be associated with the here described genotype of congenital sialidosis. Severely dilated coronary arteries, excessive retinal vascular tortuosity and an erythematous macular rash might be associated features of congenital sialidosis.

  10. Positive Selection on Hemagglutinin and Neuraminidase Genes of H1N1 Influenza Viruses

    LENUS (Irish Health Repository)

    Li, Wenfu

    2011-04-21

    Abstract Background Since its emergence in March 2009, the pandemic 2009 H1N1 influenza A virus has posed a serious threat to public health. To trace the evolutionary path of these new pathogens, we performed a selection-pressure analysis of a large number of hemagglutinin (HA) and neuraminidase (NA) gene sequences of H1N1 influenza viruses from different hosts. Results Phylogenetic analysis revealed that both HA and NA genes have evolved into five distinct clusters, with further analyses indicating that the pandemic 2009 strains have experienced the strongest positive selection. We also found evidence of strong selection acting on the seasonal human H1N1 isolates. However, swine viruses from North America and Eurasia were under weak positive selection, while there was no significant evidence of positive selection acting on the avian isolates. A site-by-site analysis revealed that the positively selected sites were located in both of the cleaved products of HA (HA1 and HA2), as well as NA. In addition, the pandemic 2009 strains were subject to differential selection pressures compared to seasonal human, North American swine and Eurasian swine H1N1 viruses. Conclusions Most of these positively and\\/or differentially selected sites were situated in the B-cell and\\/or T-cell antigenic regions, suggesting that selection at these sites might be responsible for the antigenic variation of the viruses. Moreover, some sites were also associated with glycosylation and receptor-binding ability. Thus, selection at these positions might have helped the pandemic 2009 H1N1 viruses to adapt to the new hosts after they were introduced from pigs to humans. Positive selection on position 274 of NA protein, associated with drug resistance, might account for the prevalence of drug-resistant variants of seasonal human H1N1 influenza viruses, but there was no evidence that positive selection was responsible for the spread of the drug resistance of the pandemic H1N1 strains.

  11. Simultaneous detection of hemagglutinin and neuraminidase genes of novel influenza A (H7N9) by duplex real-time reverse transcription polymerase chain reaction.

    Science.gov (United States)

    Li, Yan; Wu, Tao; Qi, Xian; Ge, Yiyue; Guo, Xiling; Wu, Bin; Yu, Huiyan; Zhu, Yefei; Shi, Zhiyang; Wang, Hua; Cui, Lunbiao; Zhou, Minghao

    2013-12-01

    A novel reassortant influenza A (H7N9) virus emerged recently in China. In this study, a duplex real-time reverse transcription polymerase chain reaction (rRT-PCR) assay was developed for the simultaneous detection of hemagglutinin (HA) and neuraminidase (NA) genes of H7N9 influenza viruses. The sensitivity of the assay was determined to be 10 RNA copies per reaction for both HA and NA genes. No cross-reactivity was observed with other influenza virus subtypes or respiratory tract viruses. One hundred and forty-six clinical and environmental specimens were tested and compared with reference methods and were found to be consistent. The assay is suitable for large-scale screening due to short turnaround times and high specificity, sensitivity, and reproducibility. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. A Viable Recombinant Rhabdovirus Lacking Its Glycoprotein Gene and Expressing Influenza Virus Hemagglutinin and Neuraminidase Is a Potent Influenza Vaccine

    Science.gov (United States)

    Ryder, Alex B.; Buonocore, Linda; Vogel, Leatrice; Nachbagauer, Raffael; Krammer, Florian

    2014-01-01

    ABSTRACT The emergence of novel influenza viruses that cause devastating human disease is an ongoing threat and serves as an impetus for the continued development of novel approaches to influenza vaccines. Influenza vaccine development has traditionally focused on producing humoral and/or cell-mediated immunity, often against the viral surface glycoproteins hemagglutinin (HA) and neuraminidase (NA). Here, we describe a new vaccine candidate that utilizes a replication-defective vesicular stomatitis virus (VSV) vector backbone that lacks the native G surface glycoprotein gene (VSVΔG). The expression of the H5 HA of an H5N1 highly pathogenic avian influenza virus (HPAIV), A/Vietnam/1203/04 (VN1203), and the NA of the mouse-adapted H1N1 influenza virus A/Puerto Rico/8/34 (PR8) in the VSVΔG vector restored the ability of the recombinant virus to replicate in cell culture, without the requirement for the addition of trypsin. We show here that this recombinant virus vaccine candidate was nonpathogenic in mice when given by either the intramuscular or intranasal route of immunization and that the in vivo replication of VSVΔG-H5N1 is profoundly attenuated. This recombinant virus also provided protection against lethal H5N1 infection after a single dose. This novel approach to vaccination against HPAIVs may be widely applicable to other emerging strains of influenza virus. IMPORTANCE Preparation for a potentially catastrophic influenza pandemic requires novel influenza vaccines that are safe, can be produced and administered quickly, and are effective, both soon after administration and for a long duration. We have created a new influenza vaccine that utilizes an attenuated vesicular stomatitis virus (VSV) vector, to deliver and express influenza virus proteins against which vaccinated animals develop potent antibody responses. The influenza virus hemagglutinin and neuraminidase proteins, expressed on the surface of VSV particles, allowed this vaccine to grow in cell

  13. A viable recombinant rhabdovirus lacking its glycoprotein gene and expressing influenza virus hemagglutinin and neuraminidase is a potent influenza vaccine.

    Science.gov (United States)

    Ryder, Alex B; Buonocore, Linda; Vogel, Leatrice; Nachbagauer, Raffael; Krammer, Florian; Rose, John K

    2015-03-01

    The emergence of novel influenza viruses that cause devastating human disease is an ongoing threat and serves as an impetus for the continued development of novel approaches to influenza vaccines. Influenza vaccine development has traditionally focused on producing humoral and/or cell-mediated immunity, often against the viral surface glycoproteins hemagglutinin (HA) and neuraminidase (NA). Here, we describe a new vaccine candidate that utilizes a replication-defective vesicular stomatitis virus (VSV) vector backbone that lacks the native G surface glycoprotein gene (VSVΔG). The expression of the H5 HA of an H5N1 highly pathogenic avian influenza virus (HPAIV), A/Vietnam/1203/04 (VN1203), and the NA of the mouse-adapted H1N1 influenza virus A/Puerto Rico/8/34 (PR8) in the VSVΔG vector restored the ability of the recombinant virus to replicate in cell culture, without the requirement for the addition of trypsin. We show here that this recombinant virus vaccine candidate was nonpathogenic in mice when given by either the intramuscular or intranasal route of immunization and that the in vivo replication of VSVΔG-H5N1 is profoundly attenuated. This recombinant virus also provided protection against lethal H5N1 infection after a single dose. This novel approach to vaccination against HPAIVs may be widely applicable to other emerging strains of influenza virus. Preparation for a potentially catastrophic influenza pandemic requires novel influenza vaccines that are safe, can be produced and administered quickly, and are effective, both soon after administration and for a long duration. We have created a new influenza vaccine that utilizes an attenuated vesicular stomatitis virus (VSV) vector, to deliver and express influenza virus proteins against which vaccinated animals develop potent antibody responses. The influenza virus hemagglutinin and neuraminidase proteins, expressed on the surface of VSV particles, allowed this vaccine to grow in cell culture and induced a

  14. Characterization of the neuraminidase genes from human influenza A viruses circulating in Iran from 2010 to 2015.

    Science.gov (United States)

    Moasser, Elham; Behzadian, Farida; Moattari, Afagh; Fotouhi, Fatemeh; Zaraket, Hassan

    2017-10-31

    Characterization of influenza viruses is critical for detection of new emerging variants. Herein, we analyzed the genetic diversity and drug susceptibility of the neuraminidase gene (NAs) expressed by influenza A/H1N1pdm09 and A/H3N2 viruses circulating in Iran from 2010 to 2015. We genetically analyzed the NAs of 38 influenza A/H1N1pdm09 and 35 A/H3N2 isolates. The Iranian A/H1N1pdm09 viruses belonged to seven genogroups/subgenogroups, with the dominant groups being genogroups 6B and 6C. The A/H3N2 isolates fell into six gneogroups/subgenogroups, with the dominant genogroups being 3C and 3C.2a. The most common mutations detected among the A/H1N1pdm09 viruses included N44S, V106I, N200S, and N248D. All H1N1pdm09 viruses were genetically susceptible to the NAIs. However, one A/H1N1pdm09 virus from the 2013-2014 season possessed an NA-S247N mutation, which reduces the susceptibility to oseltamivir. In case of H3N2, none of the analyzed Iranian strains carried a substitution that might affect its susceptibility to NAIs. The ongoing evolution of influenza viruses and the detect of influenza viruses with reduced susceptibility to NAIs warrants continuous monitoring of the circulating strains.

  15. Cloning of neuraminidase (NA) gene and identification of its antiviral ...

    African Journals Online (AJOL)

    user

    2012-06-12

    Jun 12, 2012 ... characters of isolates of Pasteurella multocida obtained from back‐yard poultry and from two outbreaks of avian cholera in avifauna in Denmark. Avian Pathol., 27(4): 373-381. Cong YL, Liu JH (2008). Molecular mechanism of the role of hemagglutinin of influenza virus in interspecies transmission. Chin. J.

  16. Large-scale FMO-MP3 calculations on the surface proteins of influenza virus, hemagglutinin (HA) and neuraminidase (NA)

    Science.gov (United States)

    Mochizuki, Yuji; Yamashita, Katsumi; Fukuzawa, Kaori; Takematsu, Kazutomo; Watanabe, Hirofumi; Taguchi, Naoki; Okiyama, Yoshio; Tsuboi, Misako; Nakano, Tatsuya; Tanaka, Shigenori

    2010-06-01

    Two proteins on the influenza virus surface have been well known. One is hemagglutinin (HA) associated with the infection to cells. The fragment molecular orbital (FMO) calculations were performed on a complex consisting of HA trimer and two Fab-fragments at the third-order Møller-Plesset perturbation (MP3) level. The numbers of residues and 6-31G basis functions were 2351 and 201276, and thus a massively parallel-vector computer was utilized to accelerate the processing. This FMO-MP3 job was completed in 5.8 h with 1024 processors. Another protein is neuraminidase (NA) involved in the escape from infected cells. The FMO-MP3 calculation was also applied to analyze the interactions between oseltamivir and surrounding residues in pharmacophore.

  17. [High-yield reassortant virus containing hemagglutinin and neuraminidase genes of pandemic influenza A/Moscowl/01/2009 (H1N1) virus].

    Science.gov (United States)

    Ignat'eva, A V; Rudneva, I A; Timofeeva, T A; Shilov, A A; Zaberezhnyĭ, A D; Aliper, T I; Kaverin, N V; L'vov, D K

    2011-01-01

    The crossing of influenza A/Moscow/01/2009 (H1N1) virus and reassortant strain X31 (H3N2) containing the genes of internal and non-structural proteins of A/Puerto Rico/8/34 (H1N1) strain gave rise to reassortant virus ReM8. The reassortant contained hemagglutinin (HA) and neuraminidase (NA) genes of pandemic 2009 influenza virus and 6 genes of high-yield A/Puerto Rico/8/34 (H1N1) strain. The reassortant ReM8 produced higher yields in the embryonated chicken eggs than the parent pandemic virus, as suggested by infectivity and HA activity titration as well as by ELISA and the measurement of HA protein content by scanning electrophoresis in polyacrylamide gel slabs. High immunogenicity of ReM8 reassortant was demonstrated by immune protection studies in mice. The reassortant virus ReM8 is suitable as a candidate strain for the production of inactivated and subunit influenza vaccines.

  18. The I427T neuraminidase (NA) substitution, located outside the NA active site of an influenza A(H1N1)pdm09 variant with reduced susceptibility to NA inhibitors, alters NA properties and impairs viral fitness.

    Science.gov (United States)

    Tu, Véronique; Abed, Yacine; Barbeau, Xavier; Carbonneau, Julie; Fage, Clément; Lagüe, Patrick; Boivin, Guy

    2017-01-01

    Emergence of pan neuraminidase inhibitor (NAI)-resistant variants constitutes a serious clinical concern. An influenza A(H1N1)pdm09 variant containing the I427T/Q313R neuraminidase (NA) substitutions was previously identified in a surveillance study. Although these changes are not part of the NA active site, the variant showed reduced susceptibility to many NAIs. In this study, we investigated the mechanism of resistance for the I427T/Q313R substitution and its impact on the NA enzyme and viral fitness. Recombinant wild-type (WT), I427T/Q313R and I427T A(H1N1)pdm09 viruses were generated by reverse genetics and tested for their drug susceptibilities, enzymatic properties and replication kinetics in vitro as well as their virulence in mice. Molecular dynamics (MD) simulations were performed for NA structural analysis. The I427T substitution, which was responsible for the resistance phenotype observed in the double (I427T/Q313R) mutant, induced 17-, 56-, 7-, and 14-fold increases in IC50 values against oseltamivir, zanamivir, peramivir and laninamivir, respectively. The I427T substitution alone or combined to Q313R significantly reduced NA affinity. The I427T/Q313R and to a lesser extent I427T recombinant viruses displayed reduced viral titers vs WT in vitro. In experimentally-infected mice, the mortality rates were 62.5%, 0% and 14.3% for the WT, I417T/Q313R and I427T viruses, respectively. There were about 2.5- and 2-Log reductions in mean lung viral titers on day 5 post-infection for the I427T/Q313R and I427T mutants, respectively, compared to WT. Results from simulations revealed that the I427T change indirectly altered the stability of the catalytic R368 residue of the NA enzyme causing its reduced binding to the substrate/inhibitor. This study demonstrates that the I427T/Q313R mutant, not only alters NAI susceptibility but also compromises NA properties and viral fitness, which could explain its infrequent detection in clinic. Copyright © 2016 Elsevier B

  19. [Accessing the features of surface neuraminidase (N1) of influenza A virus presenting on the platforms for anti-NA Abs screening].

    Science.gov (United States)

    Huang, Lan; Qin, Kun; Zhou, Jian-fang; Shu, Yue-long; Wei, Hong

    2013-02-01

    To understand if the Neuraminidase (N1) of Influenza A virus at the surface of yeast-displaying system, eukaryotic expression system and the infected cells could be used for anti-NA Abs screening, their activities and bindings to five candidate Abs were assayed. The surface NA expression was obtained by transfecting by recombinant NA constructors with specific tag-labels or live virus infection. The functional activity was measured by the fluorescent assay. Their bindings to the Abs were detected by flow cytometry. The surface NAs presenting on the yeast-displaying system and eukaryotic expression system exhibited functional NA activities as the NA at the surface of virus-infected cells which showed affinities to Ab1, 4, and 5. The same bindings to Abl and 5 were found in the surface NA expressed by eukaryotic expression system while minor binding was observed in the yeast displayed-NA. The epitopes of yeast-displayed NA may be different from the NAs present at eukaryotic expression system and the infected cells which more likely suitable for the screening of anti-NA Abs.

  20. SYBR green-based real-time reverse transcription-PCR for typing and subtyping of all hemagglutinin and neuraminidase genes of avian influenza viruses and comparison to standard serological subtyping tests

    Science.gov (United States)

    Tsukamoto, K.; Javier, P.C.; Shishido, M.; Noguchi, D.; Pearce, J.; Kang, H.-M.; Jeong, O.M.; Lee, Y.-J.; Nakanishi, K.; Ashizawa, T.

    2012-01-01

    Continuing outbreaks of H5N1 highly pathogenic (HP) avian influenza virus (AIV) infections of wild birds and poultry worldwide emphasize the need for global surveillance of wild birds. To support the future surveillance activities, we developed a SYBR green-based, real-time reverse transcriptase PCR (rRT-PCR) for detecting nucleoprotein (NP) genes and subtyping 16 hemagglutinin (HA) and 9 neuraminidase (NA) genes simultaneously. Primers were improved by focusing on Eurasian or North American lineage genes; the number of mixed-base positions per primer was set to five or fewer, and the concentration of each primer set was optimized empirically. Also, 30 cycles of amplification of 1:10 dilutions of cDNAs from cultured viruses effectively reduced minor cross- or nonspecific reactions. Under these conditions, 346 HA and 345 NA genes of 349 AIVs were detected, with average sensitivities of NP, HA, and NA genes of 10 1.5, 10 2.3, and 10 3.1 50% egg infective doses, respectively. Utility of rRT-PCR for subtyping AIVs was compared with that of current standard serological tests by using 104 recent migratory duck virus isolates. As a result, all HA genes and 99% of the NA genes were genetically subtyped, while only 45% of HA genes and 74% of NA genes were serologically subtyped. Additionally, direct subtyping of AIVs in fecal samples was possible by 40 cycles of amplification: approximately 70% of HA and NA genes of NP gene-positive samples were successfully subtyped. This validation study indicates that rRT-PCR with optimized primers and reaction conditions is a powerful tool for subtyping varied AIVs in clinical and cultured samples. Copyright ?? 2012, American Society for Microbiology. All Rights Reserved.

  1. A complete analysis of HA and NA genes of influenza A viruses.

    LENUS (Irish Health Repository)

    Shi, Weifeng

    2010-12-01

    More and more nucleotide sequences of type A influenza virus are available in public databases. Although these sequences have been the focus of many molecular epidemiological and phylogenetic analyses, most studies only deal with a few representative sequences. In this paper, we present a complete analysis of all Haemagglutinin (HA) and Neuraminidase (NA) gene sequences available to allow large scale analyses of the evolution and epidemiology of type A influenza.

  2. Antiviral resistance due to deletion in the neuraminidase gene and defective interfering-like viral polymerase basic 2 RNA of influenza A virus subtype H3N2

    DEFF Research Database (Denmark)

    Trebbien, Ramona; Christiansen, Claus Bohn; Fischer, Thea Kølsen

    2018-01-01

    two major out-of-frame deletions in the polymerase basic 2 (PB2) gene, indicating defective interfering-like viral RNA. Conclusions: The viruses harboring the 245–248 deletion in the neuraminidase gene were still present after discontinuation of oseltamivir treatment and passages in cell cultures...... to zanamivir. Nine days after discontinuation of oseltamivir treatment the deleted H3N2 virus was still present in the patient. After three passages of the deleted virus in cell culture, the deletion was retained. Several variant mutations appeared in the other genes of the H3N2 virus, where most striking were...

  3. Improved immunogenicity of Newcastle disease virus inactivated vaccine following DNA vaccination using Newcastle disease virus hemagglutinin-neuraminidase and fusion protein genes.

    Science.gov (United States)

    Firouzamandi, Masoumeh; Moeini, Hassan; Hosseini, Davood; Bejo, Mohd Hair; Omar, Abdul Rahman; Mehrbod, Parvaneh; Ideris, Aini

    2016-03-01

    The present study describes the development of DNA vaccines using the hemagglutinin-neuraminidase (HN) and fusion (F) genes from AF2240 Newcastle disease virus strain, namely pIRES/HN, pIRES/F and pIRES-F/HN. Transient expression analysis of the constructs in Vero cells revealed the successful expression of gene inserts in vitro. Moreover, in vivo experiments showed that single vaccination with the constructed plasmid DNA (pDNA) followed by a boost with inactivated vaccine induced a significant difference in enzyme-linked immunosorbent assay antibody levels (p inactivated vaccine alone. Taken together, these results indicated that recombinant pDNA could be used to increase the efficacy of the inactivated vaccine immunization procedure.

  4. Purification and renaturation of membrane neuraminidase from Haemophilus parasuis.

    Science.gov (United States)

    Lichtensteiger, Carol A; Vimr, Eric R

    2003-05-02

    Haemophilus parasuis, which causes polyserositis, polysynovitis, meningitis, septicemia, and pneumonia in pigs, has emerged as an increasing problem in modern swine production systems. Co-factors for and the pathogenesis of H. parasuis disease are not defined. One of the potential virulence factors of H. parasuis is its neuraminidase (sialidase). While purifying the H. parasuis neuraminidase from the membrane fraction, we developed a protocol to renature enzymatic activity after enzyme preparations were resolved electrophorectically in denaturing polyacrylamide gels. The H. parasuis neuraminidase co-resolved with recombinant neuraminidase of Vibrio cholera; thus its apparent molecular mass is 82 kilodalton (kDa). The H. parasuis neuraminidase was associated with the membrane fraction and the purification protocol removed over 99% of the H. parasuis cell protein while retaining over 90% of the neuraminidase activity. Purified protein will provide another avenue to clone the neuraminidase gene that has been refractory to cloning and the protocol will be a means to purify recombinant protein. Copyright 2003 Elsevier Science B.V.

  5. Regulation of neuraminidase expression in Streptococcus pneumoniae

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    Gualdi Luciana

    2012-09-01

    Full Text Available Abstract Background Sialic acid (N-acetylneuraminic acid; NeuNAc is one of the most important carbohydrates for Streptococcus pneumoniae due of its role as a carbon and energy source, receptor for adhesion and invasion and molecular signal for promotion of biofilm formation, nasopharyngeal carriage and invasion of the lung. Results In this work, NeuNAc and its metabolic derivative N-acetyl mannosamine (ManNAc were used to analyze regulatory mechanisms of the neuraminidase locus expression. Genomic and metabolic comparison to Streptococcus mitis, Streptococcus oralis, Streptococcus gordonii and Streptococcus sanguinis elucidates the metabolic association of the two amino sugars to different parts of the locus coding for the two main pneumococcal neuraminidases and confirms the substrate specificity of the respective ABC transporters. Quantitative gene expression analysis shows repression of the locus by glucose and induction of all predicted transcriptional units by ManNAc and NeuNAc, each inducing with higher efficiency the operon encoding for the transporter with higher specificity for the respective amino sugar. Cytofluorimetric analysis demonstrated enhanced surface exposure of NanA on pneumococci grown in NeuNAc and ManNAc and an activity assay allowed to quantify approximately twelve times as much neuraminidase activity on induced cells as opposed to glucose grown cells. Conclusions The present data increase the understanding of metabolic regulation of the nanAB locus and indicate that experiments aimed at the elucidation of the relevance of neuraminidases in pneumococcal virulence should possibly not be carried out on bacteria grown in glucose containing media.

  6. Optimizing Viral Protein Yield of Influenza Virus Strain A/Vietnam/1203/2004 by Modification of the Neuraminidase Gene▿

    OpenAIRE

    Adamo, Joan E.; Liu, Teresa; Schmeisser, Falko; Ye, Zhiping

    2009-01-01

    The preparation of high-yield prepandemic influenza virus H5N1 strains has presented a challenge to both researchers and vaccine manufacturers. The reasons for the relatively low yield of the H5N1 strains are not fully understood, but it might be partially dependent on the interactions between the hemagglutinin (HA) or neuraminidase (NA) surface glycoprotein and other influenza virus proteins. In this study, we have constructed chimeras between the A/Puerto Rico/8/34 (PR8) NA gene and the A/V...

  7. Inhibition of neuraminidase by Ganoderma triterpenoids and implications for neuraminidase inhibitor design

    Science.gov (United States)

    Zhu, Qinchang; Bang, Tran Hai; Ohnuki, Koichiro; Sawai, Takashi; Sawai, Ken; Shimizu, Kuniyoshi

    2015-01-01

    Neuraminidase (NA) inhibitors are the dominant antiviral drugs for treating influenza in the clinic. Increasing prevalence of drug resistance makes the discovery of new NA inhibitors a high priority. Thirty-one triterpenoids from the medicinal mushroom Ganoderma lingzhi were analyzed in an in vitro NA inhibition assay, leading to the discovery of ganoderic acid T-Q and TR as two inhibitors of H5N1 and H1N1 NAs. Structure-activity relationship studies revealed that the corresponding triterpenoid structure is a potential scaffold for the design of NA inhibitors. Using these triterpenoids as probes we found, through further in silico docking and interaction analysis, that interactions with the amino-acid residues Arg292 and/or Glu119 of NA are critical for the inhibition of H5N1 and H1N1. These findings should prove valuable for the design and development of NA inhibitors. PMID:26307417

  8. Homology modelling and insilico analysis of neuraminidase protein in H1N1 Influenza A virus

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    Abhilash Manohar

    2011-02-01

    Full Text Available In this work, modelling of Neuraminidase protein of Influenza A virus (A/Himeji/1/2009(H1N1 neuraminidase (NA protein was done using Modeller 9V2. Modelled structure was submitted to protein model database and could be downloaded using accession number PM0075830. The modelled protein structure was subjected to In silco analysis using various bioinformatics tools. Two anti-influenza drugs currently being used to treat infected patients are oseltamivir (Tamiflu and zanamivir (Relenza, both of which target the neuraminidase enzyme of the virus. Reports of the emergence of drug resistance make the development of new anti-influenza molecules a priority. Hence the modelled structure of H1NI Neuraminidase could be very useful for in silico analysis of potential neuraminidase inhibitors.

  9. Neuraminidase Inhibitory Activity and Constituent Characterization of Fagopyrum dibotrys

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    Xiang Zhang

    2017-11-01

    Full Text Available This study aimed to identify a new biological activity of the widely distributed species Fagopyrum dibotrys. Four F. dibotrys extracts (ethyl acetate (EA, petroleum ether (P, ethanol (E, and water (W were explored for their anti-neuraminidase (NA activity. A total of 32 compounds were identified using UHPLC-Q-Exactive Orbitrap HRMS in the EA extract, which had the best NA inhibitory effects. We used the docking data for supporting compounds’ anti-neuraminidase activity. Among them, five compounds including one flavonoid, three organic acids, and one glucoside were discovered for the first time in F. dibotrys. Docking studies and NA activity assay revealed the remarkable NA inhibitory activity of eight components in EA extract, especially rutin, hesperidin, procyanidin B2, and quercitrin. Therefore, F. dibotrys could be used to develop anti-influenza drugs.

  10. Splice donor site mutation in the lysosomal neuraminidase gene causing exon skipping and complete loss of enzyme activity in a sialidosis patient.

    Science.gov (United States)

    Penzel, R; Uhl, J; Kopitz, J; Beck, M; Otto, H F; Cantz, M

    2001-07-20

    Sialidosis is a lysosomal storage disease caused by the deficiency of alpha-N-acetylneuraminidase (NEU1; sialidase), the key enzyme for the intralysosomal catabolism of sialylated glycoconjugates. We have identified a homozygous transversion in the last intron (IVSE +1 G>C) in neu1 of a sialidosis patient. Sequencing of the truncated cDNA revealed an alternatively spliced neu1 transcript which lacks the complete sequence of exon 5. Skipping of exon 5 leads to a frameshift and results in a premature termination codon. This is the first description of an intronic point mutation causing a complete deficiency of the lysosomal neuraminidase activity.

  11. Neuraminidase-mediated haemagglutination of recent human influenza A(H3N2) viruses is determined by arginine 150 flanking the neuraminidase catalytic site.

    Science.gov (United States)

    Mögling, Ramona; Richard, Mathilde J; Vliet, Stefan van der; Beek, Ruud van; Schrauwen, Eefje J A; Spronken, Monique I; Rimmelzwaan, Guus F; Fouchier, Ron A M

    2017-06-01

    Over the last decade, an increasing proportion of circulating human influenza A(H3N2) viruses exhibited haemagglutination activity that was sensitive to neuraminidase inhibitors. This change in haemagglutination as compared to older circulating A(H3N2) viruses prompted an investigation of the underlying molecular basis. Recent human influenza A(H3N2) viruses were found to agglutinate turkey erythrocytes in a manner that could be blocked with either oseltamivir or neuraminidase-specific antisera, indicating that agglutination was driven by neuraminidase, with a low or negligible contribution of haemagglutinin. Using representative virus recombinants it was shown that the haemagglutinin of a recent A(H3N2) virus indeed had decreased activity to agglutinate turkey erythrocytes, while its neuraminidase displayed increased haemagglutinating activity. Viruses with chimeric and mutant neuraminidases were used to identify the amino acid substitution histidine to arginine at position 150 flanking the neuraminidase catalytic site as the determinant of this neuraminidase-mediated haemagglutination. An analysis of publicly available neuraminidase gene sequences showed that viruses with histidine at position 150 were rapidly replaced by viruses with arginine at this position between 2005 and 2008, in agreement with the phenotypic data. As a consequence of neuraminidase-mediated haemagglutination of recent A(H3N2) viruses and poor haemagglutination via haemagglutinin, haemagglutination inhibition assays with A(H3N2) antisera are no longer useful to characterize the antigenic properties of the haemagglutinin of these viruses for vaccine strain selection purposes. Continuous monitoring of the evolution of these viruses and potential consequences for vaccine strain selection remains important.

  12. Catalytically defective ganglioside neuraminidase in mucolipidosis IV

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    Ben-Yoseph, Y.; Momoi, T.; Hahn, L.C.; Nadler, H.L. (Wayne State Univ., Detroit, MI (USA))

    1982-01-01

    Cultured skin fibroblasts from patients with mucolipidosis IV were found to be deficient in neuraminidase activity toward GDsub(la) and GDsub(lb) gangliosides radiolabelled in C/sub 3/ and C/sub 7/ analogs of their sialic acid residues. Neuraminidase activities toward 4-methylumbelliferyl-N-acetyl-neuraminic acid, neuraminlactose, and radiolabelled neuraminlactitol, fetuin and ..cap alpha../sub 1/-acid glycoprotein were within the range of normal controls. Fibroblasts from parents of patients with mucolipidosis IV demonstrated intermediate levels of ganglioside neuraminidase activity and normal levels of glycoprotein neuraminidase activity. The redidual acidic neuraminidase activity toward GDsub(1a) ganglioside in the patients' fibroblasts did not differ from that of controls in its pH optimum and thermostability, but had an abnormal apparent Ksub(m) which was about 18 times higher than that of the normal enzyme. These findings suggest that mucolipidosis IV is a ganglioside sialidosis due to a catalytically defective ganglioside neuraminidase.

  13. Detailed genetic analysis of hemagglutinin-neuraminidase glycoprotein gene in human parainfluenza virus type 1 isolates from patients with acute respiratory infection between 2002 and 2009 in Yamagata prefecture, Japan

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    Mizuta Katsumi

    2011-12-01

    Full Text Available Abstract Background Human parainfluenza virus type 1 (HPIV1 causes various acute respiratory infections (ARI. Hemagglutinin-neuraminidase (HN glycoprotein of HPIV1 is a major antigen. However, the molecular epidemiology and genetic characteristics of such ARI are not exactly known. Recent studies suggested that a phylogenetic analysis tool, namely the maximum likelihood (ML method, may be applied to estimate the evolutionary time scale of various viruses. Thus, we conducted detailed genetic analyses including homology analysis, phylogenetic analysis (using both the neighbor joining (NJ and ML methods, and analysis of the pairwise distances of HN gene in HPIV1 isolated from patients with ARI in Yamagata prefecture, Japan. Results A few substitutions of nucleotides in the second binding site of HN gene were observed among the present isolates. The strains were classified into two major clusters in the phylogenetic tree by the NJ method. Another phylogenetic tree constructed by the ML method showed that the strains diversified in the late 1980s. No positively selected sites were found in the present strains. Moreover, the pairwise distance among the present isolates was relatively short. Conclusions The evolution of HN gene in the present HPIV1 isolates was relatively slow. The ML method may be a useful phylogenetic method to estimate the evolutionary time scale of HPIV and other viruses.

  14. Antibody against Microbial Neuraminidases Recognizes Human Sialidase 3 (NEU3: the Neuraminidase/Sialidase Superfamily Revisited

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    Chiguang Feng

    2017-06-01

    Full Text Available Neuraminidases (NAs are critical virulence factors for several microbial pathogens. With a highly conserved catalytic domain, a microbial NA “superfamily” has been proposed. We previously reported that murine polymorphonuclear leukocyte (PMN sialidase activity was important in leukocyte trafficking to inflamed sites and that antibodies to Clostridium perfringens NA recognized a cell surface molecule(s, presumed to be a sialidase of eukaryotic origin on interleukin-8-stimulated human and murine PMNs. These antibodies also inhibited cell sialidase activity both in vitro and, in the latter instance, in vivo. We therefore hypothesized that mammalian sialidases share structural homology and epitopes with microbial NAs. We now report that antibodies to one of the isoforms of C. perfringens NA, as well as anti-influenza virus NA serum, recognize human NEU3 but not NEU1 and that antibodies to C. perfringens NA inhibit NEU3 enzymatic activity. We conclude that the previously described microbial NA superfamily extends to human sialidases. Strategies designed to therapeutically inhibit microbial NA may need to consider potential compromising effects on human sialidases, particularly those expressed in cells of the immune system.

  15. Screening for Neuraminidase Inhibitory Activity in Traditional Chinese Medicines Used to Treat Influenza

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    Xian-Ying Yang

    2016-08-01

    Full Text Available Objective: To screen for influenza virus neuraminidase inhibition and to provide a reference for the clinical treatment of influenza using traditional Chinese medicines (TCM. In this study, 421 crude extracts (solubilized with petroleum ether, ethanol, ethyl acetate, and aqueous solvents were obtained from 113 TCM. The medicine extracts were then reacted with oseltamivir, using 2’-(4-methylumbelliferyl-α-D-N-acetylneuraminic acid (MUNANA as the substrate, to determine influenza virus neuraminidase activity using a standard fluorimetric assay. It was found that Chinese medicine extracts from Pyrola calliantha, Cynanchum wilfordii, Balanophora involucrata and Paeonia delavayi significantly inhibited neuraminidase activity at a concentration of 40 μg/mL. Dose-dependent inhibitory assays also revealed significant inhibition. The IC50 range of the TCM extracts for influenza virus neuraminidase was approximately 12.66–34.85 μg/mL, respectively. Some Chinese medicines have clear anti-influenza viral effects that may play an important role in the treatment of influenza through the inhibition of viral neuraminidase. The results of this study demonstrated that plant medicines can serve as a useful source of neuraminidase (NA inhibitors and further investigation into the pharmacologic activities of these extracts is warranted.

  16. The M segment of the 2009 pandemic influenza virus confers increased neuraminidase activity, filamentous morphology, and efficient contact transmissibility to A/Puerto Rico/8/1934-based reassortant viruses.

    Science.gov (United States)

    Campbell, Patricia J; Danzy, Shamika; Kyriakis, Constantinos S; Deymier, Martin J; Lowen, Anice C; Steel, John

    2014-04-01

    The 2009 H1N1 lineage represented the first detection of a novel, highly transmissible influenza A virus genotype: six gene segments originated from the North American triple-reassortant swine lineage, and two segments, NA and M, derived from the Eurasian avian-like swine lineage. As neither parental lineage transmits efficiently between humans, the adaptations and mechanisms underlying the pandemic spread of the swine-origin 2009 strain are not clear. To help identify determinants of transmission, we used reverse genetics to introduce gene segments of an early pandemic isolate, A/Netherlands/602/2009 [H1N1] (NL602), into the background of A/Puerto Rico/8/1934 [H1N1] (PR8) and evaluated the resultant viruses in a guinea pig transmission model. Whereas the NL602 virus spread efficiently, the PR8 virus did not transmit. Swapping of the HA, NA, and M segments of NL602 into the PR8 background yielded a virus with indistinguishable contact transmissibility to the wild-type pandemic strain. Consistent with earlier reports, the pandemic M segment alone accounted for much of the improvement in transmission. To aid in understanding how the M segment might affect transmission, we evaluated neuraminidase activity and virion morphology of reassortant viruses. Transmission was found to correlate with higher neuraminidase activity and a more filamentous morphology. Importantly, we found that introduction of the pandemic M segment alone resulted in an increase in the neuraminidase activity of two pairs of otherwise isogenic PR8-based viruses. Thus, our data demonstrate the surprising result that functions encoded by the influenza A virus M segment impact neuraminidase activity and, perhaps through this mechanism, have a potent effect on transmissibility. Our work uncovers a previously unappreciated mechanism through which the influenza A virus M segment can alter the receptor-destroying activity of an influenza virus. Concomitant with changes to neuraminidase activity, the M

  17. Neuraminidase deficiency: case report and review of the phenotype.

    Science.gov (United States)

    Young, I D; Young, E P; Mossman, J; Fielder, A R; Moore, J R

    1987-01-01

    A 12 year old boy with neuraminidase deficiency (sialidosis, mucolipidosis I) is described. His clinical features included coarse facies, cherry red spot, ataxia, myoclonus, and dysotosis multiplex. The level of neuraminidase activity in cultured fibroblasts was very low and intermediate levels were observed in both parents. The clinical disorders associated with neuraminidase deficiency are reviewed. Images PMID:3585942

  18. Influenza A Virus with a Human-Like N2 Gene Is Circulating in Pigs

    DEFF Research Database (Denmark)

    Breum, Solvej Østergaard; Hjulsager, Charlotte Kristiane; Trebbien, Ramona

    2013-01-01

    A novel reassortant influenza A virus, H1avN2hu, has been found in Danish swine. The virus contains an H1 gene similar to the hemagglutinin (HA) gene of H1N1 avian-like swine viruses and an N2 gene most closely related to the neuraminidase (NA) gene of human H3N2 viruses from the mid-1990s....

  19. Crystal Structures of Respiratory Pathogen Neuraminidases

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    Hsiao, Y.; Parker, D; Ratner, A; Prince, A; Tong, L

    2009-01-01

    Currently there is pressing need to develop novel therapeutic agents for the treatment of infections by the human respiratory pathogens Pseudomonas aeruginosa and Streptococcus pneumoniae. The neuraminidases of these pathogens are important for host colonization in animal models of infection and are attractive targets for drug discovery. To aid in the development of inhibitors against these neuraminidases, we have determined the crystal structures of the P. aeruginosa enzyme NanPs and S. pneumoniae enzyme NanA at 1.6 and 1.7 {angstrom} resolution, respectively. In situ proteolysis with trypsin was essential for the crystallization of our recombinant NanA. The active site regions of the two enzymes are strikingly different. NanA contains a deep pocket that is similar to that in canonical neuraminidases, while the NanPs active site is much more open. The comparative studies suggest that NanPs may not be a classical neuraminidase, and may have distinct natural substrates and physiological functions. This work represents an important step in the development of drugs to prevent respiratory tract colonization by these two pathogens.

  20. Novel mutations in lysosomal neuraminidase identify functional domains and determine clinical severity in sialidosis.

    Science.gov (United States)

    Bonten, E J; Arts, W F; Beck, M; Covanis, A; Donati, M A; Parini, R; Zammarchi, E; d'Azzo, A

    2000-11-01

    Lysosomal neuraminidase is the key enzyme for the intralysosomal catabolism of sialylated glycoconjugates and is deficient in two neurodegenerative lysosomal disorders, sialidosis and galactosialidosis. Here we report the identification of eight novel mutations in the neuraminidase gene of 11 sialidosis patients with various degrees of disease penetrance. Comparison of the primary structure of human neuraminidase with the primary and tertiary structures of bacterial sialidases indicated that most of the single amino acid substitutions occurred in functional motifs or conserved residues. On the basis of the subcellular distribution and residual catalytic activity of the mutant neuraminidases we assigned the mutant proteins to three groups: (i) catalytically inactive and not lysosomal; (ii) catalytically inactive, but localized in lysosome; and (iii) catalytically active and lysosomal. In general, there was a close correlation between the residual activity of the mutant enzymes and the clinical severity of disease. Patients with the severe infantile type II disease had mutations from group I, whereas patients with a mild form of type I disease had at least one mutation from group III. Mutations from the second group were mainly found in juvenile type II patients with intermediate clinical severity. Overall, our findings explain the clinical heterogeneity observed in sialidosis and may help in the assignment of existing or new allelic combinations to specific phenotypes.

  1. Neuraminidase activity provides a practical read-out for a high throughput influenza antiviral screening assay

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    Wu Meng

    2008-09-01

    Full Text Available Abstract Background The emergence of influenza strains that are resistant to commonly used antivirals has highlighted the need to develop new compounds that target viral gene products or host mechanisms that are essential for effective virus replication. Existing assays to identify potential antiviral compounds often use high throughput screening assays that target specific viral replication steps. To broaden the search for antivirals, cell-based replication assays can be performed, but these are often labor intensive and have limited throughput. Results We have adapted a traditional virus neutralization assay to develop a practical, cell-based, high throughput screening assay. This assay uses viral neuraminidase (NA as a read-out to quantify influenza replication, thereby offering an assay that is both rapid and sensitive. In addition to identification of inhibitors that target either viral or host factors, the assay allows simultaneous evaluation of drug toxicity. Antiviral activity was demonstrated for a number of known influenza inhibitors including amantadine that targets the M2 ion channel, zanamivir that targets NA, ribavirin that targets IMP dehydrogenase, and bis-indolyl maleimide that targets protein kinase A/C. Amantadine-resistant strains were identified by comparing IC50 with that of the wild-type virus. Conclusion Antivirals with specificity for a broad range of targets are easily identified in an accelerated viral inhibition assay that uses NA as a read-out of replication. This assay is suitable for high throughput screening to identify potential antivirals or can be used to identify drug-resistant influenza strains.

  2. Purification and properties of rabbit spermatozoal acrosomal neuraminidase.

    Science.gov (United States)

    Srivastava, P N; Abou-Issa, H

    1977-01-01

    Treatment of rabbit spermatozoa with 50mM-MgCl2 removes the plasma and the outer acrosomal membranes. Subsequent treatment with the detergents Hyamine 2389 and Triton X-100 solubilizes spermatozoal neuraminidase bound to the inner acrosomal membrane. The enzyme was further purified by DEAE-cellulose, Sephadex G-150 and Bio-Gel P-300 column chromato. The enzyme showed a single major band, with the possibility of some minor contaminants, on disc-gel electrophoresis. It had a specific activity of 0.37 micronmal of sialic acid released/min per mg with purified boar Cowper's-gland mucin as the substrate. The enzyme had marked specificity for 2 leads to 6'-linked sialic acid in glycoproteins. The Km of spermatozoal neuraminidase was 1.72 X 10(-6)M with Cowper's-gland mucin, 1.17 X 10(-5)M with fetuin and 8.8 X 10(-4)M with sialyl-lactose as a substrates. The Vmax. was 0.112 micronmol/min per mg with the Cowper's-gland mucin, 0.071 micronmol/min per mg with fetuin and 0.033 micronmol/min per mg with sialyl-lactose as substrate. The enzyme hydrolysed sheep submaxillary-gland mucin as readily as the Cowper's-gland mucin. The optimum of enzyme activity was at pH 5.0 on the Cowper's-gland mucin and at pH4.3 on sialyl-lactose. The enzyme activity was unaffected by 20mM-Na+ and-K+, but was inhibited by 20mM-Ca2+,-Mn2+,-Co2+ and -Cu2+. The enzyme was unstable in dilute solutions, but could be stored indefinitely freeze-dried at --20 degrees C. Images PLATE 1 PMID:66917

  3. Lactococcus lactis displayed neuraminidase confers cross protective immunity against influenza A viruses in mice.

    Science.gov (United States)

    Lei, Han; Peng, Xiaojue; Zhao, Daxian; Ouyang, Jiexiu; Jiao, Huifeng; Shu, Handing; Ge, Xinqi

    2015-02-01

    Influenza A viruses pose a serious threat to public health. Current influenza A vaccines predominantly focus on hemagglutinin (HA) and show strain-specific protection. Neuraminidase (NA) is much less studied in the context of humoral immunity against influenza A viruses. The purpose of this study is to evaluate the cross protective immunity of NA presented on Lactococcus lactis (L.lactis) surface against homologous and heterologous influenza A viruses in the mouse model. L.lactis/pNZ8110-pgsA-NA was constructed in which pgsA was used as an anchor protein. Mice vaccinated orally with L.lactis/pNZ8110-pgsA-NA could elicit significant NA-specific serum IgG and mucosa IgA antibodies, as well as neuraminidase inhibition (NI) titers. Importantly, L.lactis/pNZ8110-pgsA-NA provided 80% protection against H5N1, 60% protection against H3N2 and H1N1, respectively. These findings suggest that recombinant L.lactis/pNZ110-pgsA-NA in the absence of adjuvant via oral administration can be served as an effective vaccine candidate against diverse strains of influenza A viruses. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. The low-pH stability discovered in neuraminidase of 1918 pandemic influenza A virus enhances virus replication.

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    Tadanobu Takahashi

    Full Text Available The "Spanish" pandemic influenza A virus, which killed more than 20 million worldwide in 1918-19, is one of the serious pathogens in recorded history. Characterization of the 1918 pandemic virus reconstructed by reverse genetics showed that PB1, hemagglutinin (HA, and neuraminidase (NA genes contributed to the viral replication and virulence of the 1918 pandemic influenza virus. However, the function of the NA gene has remained unknown. Here we show that the avian-like low-pH stability of sialidase activity discovered in the 1918 pandemic virus NA contributes to the viral replication efficiency. We found that deletion of Thr at position 435 or deletion of Gly at position 455 in the 1918 pandemic virus NA was related to the low-pH stability of the sialidase activity in the 1918 pandemic virus NA by comparison with the sequences of other human N1 NAs and sialidase activity of chimeric constructs. Both amino acids were located in or near the amino acid resides that were important for stabilization of the native tetramer structure in a low-pH condition like the N2 NAs of pandemic viruses that emerged in 1957 and 1968. Two reverse-genetic viruses were generated from a genetic background of A/WSN/33 (H1N1 that included low-pH-unstable N1 NA from A/USSR/92/77 (H1N1 and its counterpart N1 NA in which sialidase activity was converted to a low-pH-stable property by a deletion and substitutions of two amino acid residues at position 435 and 455 related to the low-pH stability of the sialidase activity in 1918 NA. The mutant virus that included "Spanish Flu"-like low-pH-stable NA showed remarkable replication in comparison with the mutant virus that included low-pH-unstable N1 NA. Our results suggest that the avian-like low-pH stability of sialidase activity in the 1918 pandemic virus NA contributes to the viral replication efficiency.

  5. A contributing role for anti-neuraminidase antibodies on immunity to pandemic H1N1 2009 influenza A virus.

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    Glendie Marcelin

    Full Text Available Exposure to contemporary seasonal influenza A viruses affords partial immunity to pandemic H1N1 2009 influenza A virus (pH1N1 infection. The impact of antibodies to the neuraminidase (NA of seasonal influenza A viruses to cross-immunity against pH1N1 infection is unknown.Antibodies to the NA of different seasonal H1N1 influenza strains were tested for cross-reactivity against A/California/04/09 (pH1N1. A panel of reverse genetic (rg recombinant viruses was generated containing 7 genes of the H1N1 influenza strain A/Puerto Rico/08/34 (PR8 and the NA gene of either the pandemic H1N1 2009 strain (pH1N1 or one of the following contemporary seasonal H1N1 strains: A/Solomon/03/06 (rg Solomon or A/Brisbane/59/07 (rg Brisbane. Convalescent sera collected from mice infected with recombinant viruses were measured for cross-reactive antibodies to pH1N1 via Hemagglutinin Inhibition (HI or Enzyme-Linked Immunosorbent Assay (ELISA. The ectodomain of a recombinant NA protein from the pH1N1 strain (pNA-ecto was expressed, purified and used in ELISA to measure cross-reactive antibodies. Analysis of sera from elderly humans immunized with trivalent split-inactivated influenza (TIV seasonal vaccines prior to 2009 revealed considerable cross-reactivity to pNA-ecto. High titers of cross-reactive antibodies were detected in mice inoculated with either rg Solomon or rg Brisbane. Convalescent sera from mice inoculated with recombinant viruses were used to immunize naïve recipient Balb/c mice by passive transfer prior to challenge with pH1N1. Mice receiving rg California sera were better protected than animals receiving rg Solomon or rg Brisbane sera.The NA of contemporary seasonal H1N1 influenza strains induces a cross-reactive antibody response to pH1N1 that correlates with reduced lethality from pH1N1 challenge, albeit less efficiently than anti-pH1N1 NA antibodies. These findings demonstrate that seasonal NA antibodies contribute to but are not sufficient for cross

  6. Neuraminidase-Mediated, NKp46-Dependent Immune-Evasion Mechanism of Influenza Viruses

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    Yotam Bar-On

    2013-04-01

    Full Text Available Natural killer (NK cells play an essential role in the defense against influenza virus, one of the deadliest respiratory viruses known today. The NKp46 receptor, expressed by NK cells, is critical for controlling influenza infections, as influenza-virus-infected cells are eliminated through the recognition of the viral hemagglutinin (HA protein by NKp46. Here, we describe an immune-evasion mechanism of influenza viruses that is mediated by the neuraminidase (NA protein. By using various NA blockers, we show that NA removes sialic acid residues from NKp46 and that this leads to reduced recognition of HA. Furthermore, we provide in vivo and in vitro evidence for the existence of this NA-mediated, NKp46-dependent immune-evasion mechanism and demonstrate that NA inhibitors, which are commonly used for the treatment of influenza infections, are useful not only as blockers of virus budding but also as boosters of NKp46 recognition.

  7. Improvement of influenza vaccine strain A/Vietnam/1194/2004 (H5N1) growth with the neuraminidase packaging sequence from A/Puerto Rico/8/34.

    Science.gov (United States)

    Pan, Weiqi; Dong, Zhenyuan; Meng, Weixu; Zhang, Wei; Li, Ting; Li, Chufang; Zhang, Beiwu; Chen, Ling

    2012-02-01

    H5N1 influenza candidate vaccine viruses were developed using the "6+2" approach. The hemagglutinin (HA) and neuraminidase (NA) genes were derived from the popular H5N1 virus and the remaining six internal segments were derived from the A/Puerto Rico/8/34 strain (H1N1, PR8). However, some of these candidate strains have been reported to produce relatively low yields in vaccine manufacture. In this study, we found that the NA vRNA of the A/Vietnam/1194/2004 strain (H5N1, VN1194) was poorly packaged into recombinant viruses with a backbone of PR8 genes, which resulted in the formation of defective virions that did not include the NA vRNA in the genome. Using recombinant DNA techniques, we constructed a chimeric NA gene with the coding region of VN1194 NA flanked by the packaging signal sequence of the PR8 NA gene (41 bp form the 3' end of the vRNA and 67 bp from the 5' end). The packaging of the NA vRNA was restored to normal levels in the recombinant viruses containing the chimeric NA gene. Recombinant viruses containing the chimeric NA replicated much better in chicken embryonated eggs than viruses with the wild-type NA from VN1194. These findings suggest a novel strategy to improve in ovo growth of vaccine strains and to increase the number of vaccine doses available to save people if a pandemic were to occur.

  8. Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Zhengliang L., E-mail: Leon.wu@bio-techne.com [Bio-Techne Inc., 614 McKinley Place NE, Minneapolis, MN 55413 (United States); Zhou, Hui [Gregg Hall, UNH Glycomics Center, University of New Hampshire (United States); Ethen, Cheryl M. [Bio-Techne Inc., 614 McKinley Place NE, Minneapolis, MN 55413 (United States); Reinhold, Vernon N., E-mail: Vernon.Reinhold@unh.edu [Gregg Hall, UNH Glycomics Center, University of New Hampshire (United States)

    2016-04-29

    The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. Yet to date, the structure component responsible for its virulence is still a mystery. In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. In this report, the N-glycans from both forms were released and characterized by mass spectrometry. It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. These findings were further supported by the results obtained from in vitro incorporation of azido fucose and {sup 3}H-labeled fucose using core-6 fucosyltransferase, FUT8. In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. - Graphical abstract: Proposed mechanism for how core-fucose prohibits the tetramerization of the 1918 pandemic viral neuraminidase. Only the cross section of the stalk region with two N-linked glycans are depicted for clarity. (A) Carbohydrate–carbohydrate interaction on non-fucosylated monomer allows tetramerization. (B) Core-fucosylation disrupts the interaction and prevents the tetramerization. - Highlights: • Expressed 1918 pandemic influenza viral neuraminidase has inactive and active forms. • The inactive form contains high level of core-6 fucose, while the active form lacks such modification. • Core fucose could interfere the oligomerization of the neuraminidase and thus its activation. • This discovery may explain

  9. Molecular characterization and expression analysis of the Na+/H+ exchanger gene family in Medicago truncatula

    Science.gov (United States)

    One important mechanism plants use to cope with salinity is keeping the cytosolic Na+ concentration low by sequestering Na+ in vacuoles, a process facilitated by Na+/H+ exchangers (NHX). There are eight NHX genes (NHX1 through NHX8) identified and characterized in Arabidopsis. Bioinformatic analysis...

  10. Identification of Nocobactin NA Biosynthetic Gene Clusters in Nocardia farcinica▿ §

    OpenAIRE

    Hoshino, Yasutaka; Chiba, Kazuhiro; Ishino, Keiko; Fukai, Toshio; Igarashi, Yasuhiro; Yazawa, Katsukiyo; Mikami, Yuzuru; Ishikawa, Jun

    2010-01-01

    We identified the biosynthetic gene clusters of the siderophore nocobactin NA. The nbt clusters, which were discovered as genes highly homologous to the mycobactin biosynthesis genes by the genomic sequencing of Nocardia farcinica IFM 10152, consist of 10 genes separately located at two genomic regions. The gene organization of the nbt clusters and the predicted functions of the nbt genes, particularly the cyclization and epimerization domains, were in good agreement with the chemical structu...

  11. Radioassay method of neuraminidase towards N-acetylneuraminosyl hexasaccharides

    Energy Technology Data Exchange (ETDEWEB)

    Kuriyama, M.; Someya, F.; Yamada, T.; Miyatake, T. (Tokyo Metropolitan Research Lab. of Public Health (Japan))

    1982-02-26

    The authors have devised a sensitive method to assay for neuraminidase activities towards ..cap alpha..-(2..-->..3)-N-acetylneuraminosyl hexasaccharide and ..cap alpha..-(2..-->..6)-N-acetylneuraminosyl hexasaccharide, which were isolated from the urine of a patient with adult sialidosis with partial deficiency of ..beta..-galactosidase. Standard assay conditions for the determination of these neuraminidase activities were established and the radiolabeled reduced derivatives of these substrates were used. The fibroblast neuraminidase had its maximum activity at pH 4.0-4.2, with Ksub(m) values of 2.22 x 10/sup -3/ and 4.17 x 10/sup -3/ mol/l and Vsub(max) values of 76.9 and 28.6 nmol.mg/sup -1/ protein.h/sup -1/ towards the 2..-->..3 isomer and the 2..-->..6 isomer, respectively. Neuraminidase deficiencies were found in the fibroblasts of adult sialidosis, mucolipidosis II and III. These studies were compared with the neuraminidase activity towards ..cap alpha..-(2..-->..3)-N-acetylneuraminosyl lactose.

  12. EFFICIENCY AND SAFETY OF NEURAMINIDASE INHIBITOR OSELTAMIVIR FOR CHILDREN

    Directory of Open Access Journals (Sweden)

    A.S. Darmanyan

    2007-01-01

    Full Text Available Every year 3 to 5 million people suffer from flu. Children are most susceptible to flu. At the moment, the main methods of flu prophylaxis and treatment are flu vaccination and antivirus medications. Of the tested anti virus medications, 3 groups are most interesting and interferon inductors, м 2=channel blockers (derivative of adamantine and neuraminidase inhibitors. neuraminidase inhibitors are a brand new type of virus infection therapy — they selectively inhibit activity of flu virus neuraminidases, which restrains virus from getting into the cell, exit of virion from the cell before the reproduction cycle is over and new cells are affected. Oseltamivir — one of the new neuraminidase inhibitors — can be used for a safe and efficient flu treatment and prophylaxis among adults; yet recently this medication has been authorized for children over 1 year of age, basing upon the findings of randomized controlled surveys. Data shows that usage of oseltamivir reduces the period of diseases and severity of acute flu symptoms for children with no side diseases, as well as for children suffering from cardiovascular and bronchopulmonary system diseases; besides this medication reduces the risk of secondary flu complications.Key words: neuraminidase inhibitors, oseltamivir, flu, bird flu, antivirus medications.

  13. Analysis of the Na+/Ca2+ exchanger gene family within the phylum Nematoda.

    Directory of Open Access Journals (Sweden)

    Chao He

    Full Text Available Na+/Ca2+ exchangers are low affinity, high capacity transporters that rapidly transport calcium at the plasma membrane, mitochondrion, endoplasmic (and sarcoplasmic reticulum, and the nucleus. Na+/Ca2+ exchangers are widely expressed in diverse cell types where they contribute homeostatic balance to calcium levels. In animals, Na+/Ca2+ exchangers are divided into three groups based upon stoichiometry: Na+/Ca2+ exchangers (NCX, Na+/Ca2+/K+ exchangers (NCKX, and Ca2+/Cation exchangers (CCX. In mammals there are three NCX genes, five NCKX genes and one CCX (NCLX gene. The genome of the nematode Caenorhabditis elegans contains ten Na+/Ca2+ exchanger genes: three NCX; five CCX; and two NCKX genes. Here we set out to characterize structural and taxonomic specializations within the family of Na+/Ca2+ exchangers across the phylum Nematoda. In this analysis we identify Na+/Ca2+ exchanger genes from twelve species of nematodes and reconstruct their phylogenetic and evolutionary relationships. The most notable feature of the resulting phylogenies was the heterogeneous evolution observed within exchanger subtypes. Specifically, in the case of the CCX exchangers we did not detect members of this class in three Clade III nematodes. Within the Caenorhabditis and Pristionchus lineages we identify between three and five CCX representatives, whereas in other Clade V and also Clade IV nematode taxa we only observed a single CCX gene in each species, and in the Clade III nematode taxa that we sampled we identify NCX and NCKX encoding genes but no evidence of CCX representatives using our mining approach. We also provided re-annotation for predicted CCX gene structures from Heterorhabditis bacteriophora and Caenorhabditis japonica by RT-PCR and sequencing. Together, these findings reveal a complex picture of Na+/Ca2+ transporters in nematodes that suggest an incongruent evolutionary history of proteins that provide central control of calcium dynamics.

  14. Analysis of the Na+/Ca2+ exchanger gene family within the phylum Nematoda.

    Science.gov (United States)

    He, Chao; O'Halloran, Damien M

    2014-01-01

    Na+/Ca2+ exchangers are low affinity, high capacity transporters that rapidly transport calcium at the plasma membrane, mitochondrion, endoplasmic (and sarcoplasmic) reticulum, and the nucleus. Na+/Ca2+ exchangers are widely expressed in diverse cell types where they contribute homeostatic balance to calcium levels. In animals, Na+/Ca2+ exchangers are divided into three groups based upon stoichiometry: Na+/Ca2+ exchangers (NCX), Na+/Ca2+/K+ exchangers (NCKX), and Ca2+/Cation exchangers (CCX). In mammals there are three NCX genes, five NCKX genes and one CCX (NCLX) gene. The genome of the nematode Caenorhabditis elegans contains ten Na+/Ca2+ exchanger genes: three NCX; five CCX; and two NCKX genes. Here we set out to characterize structural and taxonomic specializations within the family of Na+/Ca2+ exchangers across the phylum Nematoda. In this analysis we identify Na+/Ca2+ exchanger genes from twelve species of nematodes and reconstruct their phylogenetic and evolutionary relationships. The most notable feature of the resulting phylogenies was the heterogeneous evolution observed within exchanger subtypes. Specifically, in the case of the CCX exchangers we did not detect members of this class in three Clade III nematodes. Within the Caenorhabditis and Pristionchus lineages we identify between three and five CCX representatives, whereas in other Clade V and also Clade IV nematode taxa we only observed a single CCX gene in each species, and in the Clade III nematode taxa that we sampled we identify NCX and NCKX encoding genes but no evidence of CCX representatives using our mining approach. We also provided re-annotation for predicted CCX gene structures from Heterorhabditis bacteriophora and Caenorhabditis japonica by RT-PCR and sequencing. Together, these findings reveal a complex picture of Na+/Ca2+ transporters in nematodes that suggest an incongruent evolutionary history of proteins that provide central control of calcium dynamics.

  15. Estimation of the neuraminidase content of influenza viruses and split-product vaccines by immunochromatography.

    Science.gov (United States)

    Tanimoto, Takeshi; Nakatsu, Ritsuko; Fuke, Isao; Ishikawa, Toyokazu; Ishibashi, Masahide; Yamanishi, Kouichi; Takahashi, Michiaki; Tamura, Shin-ichi

    2005-08-31

    The neuraminidase (NA) of the influenza virus, as well as the hemagglutinin, is the most important protective components in the vaccine. However, the NA content of the vaccine remains to be standardized because of the labile nature of this glycoprotein during various chemical treatments and storage. In the present study, the NA content of the split-product (SP) vaccine (virus treated with ether then formalin) was estimated together with that of the virus by an immunochoromatography technique using monoclonal antibodies (mAbs) to viral NA for A/Panama/2007/99 (A/Pa) (H3N2), B/Shangdong/7/97 (B/S) or A/New Caledonia/20/99 (A/NC) (H1N1) viral strains. In the new method, the NA catalytic activity of each fraction from steps of NA purification was measured as an index of NA content. The NA level of A/Pa, B/S or A/NC viral particles was estimated at 6.9+/-0.9, 7.6+/-0.8 or 8.5+/-1.7% of total viral protein (not significant difference between viral strains). The NA level of the corresponding A/Pa, B/S or A/NC vaccines was estimated at 9.6+/-1.5, 12.7+/-0.4 or 12.2+/-1.2% of the total vaccine protein (a significant difference between each strain of virus and its vaccine). These results suggest that the NA content in the N1, N2 or B type NA virus ranges from 5 to 11% of the total viral protein, and that the NA level in each split-product vaccine is 1.4- to 1.6-fold higher than that in the corresponding viral particles. They also suggest that the NA content can be estimated by the immunochoromatography technique using anti-viral NA mAbs.

  16. Structural basis for a class of nanomolar influenza A neuraminidase inhibitors

    Science.gov (United States)

    Kerry, Philip S.; Mohan, Sankar; Russell, Rupert J. M.; Bance, Nicole; Niikura, Masahiro; Pinto, B. Mario

    2013-10-01

    The influenza virus neuraminidase (NA) is essential for the virus life cycle. The rise of resistance mutations against current antiviral therapies has increased the need for the development of novel inhibitors. Recent efforts have targeted a cavity adjacent to the catalytic site (the 150-cavity) in addition to the primary catalytic subsite in order to increase specificity and reduce the likelihood of resistance. This study details structural and in vitro analyses of a class of inhibitors that bind uniquely in both subsites. Crystal structures of three inhibitors show occupation of the 150-cavity in two distinct and novel binding modes. We believe these are the first nanomolar inhibitors of NA to be characterized in this way. Furthermore, we show that one inhibitor, binding within the catalytic site, offers reduced susceptibility to known resistance mutations via increased flexibility of a pendant pentyloxy group and the ability to pivot about a strong hydrogen-bonding network.

  17. Syntheses and neuraminidase inhibitory activity of multisubstituted cyclopentane amide derivatives.

    Science.gov (United States)

    Chand, Pooran; Babu, Y Sudhakar; Bantia, Shanta; Rowland, Scott; Dehghani, Ali; Kotian, Pravin L; Hutchison, Tracy L; Ali, Shoukath; Brouillette, Wayne; El-Kattan, Yahya; Lin, Tsu-Hsing

    2004-04-08

    In further studies aimed toward identifying effective and safe inhibitors of influenza neuraminidases, we synthesized a series of multisubstituted cyclopentane amide derivatives. Amides prepared were 14 examples of N-substituted alkyl or aralkyl types from primary amines, 13 examples of the N,N-disubstituted alkyl, aralkyl, or substituted-alkyl type from secondary amines, and 12 examples from cycloaliphatic or substituted cycloaliphatic secondary amines. These compounds bearing two chiral centers, at position-1 in the ring and position-1' in the side chain attached at position 3, were tested for their ability to inhibit A and B forms of influenza neuraminidase. The 1-ethylpropylamide, diethylamide, dipropylamide, and 4-morpholinylamide showed very good inhibitory activity (IC(50) = 0.015-0.080 microM) vs the neuraminidase A form, but modest activity (IC(50) = 3.0-9.2 microM) vs the neuraminidase B form. Since the parent amides bear two chiral centers (C-1 and C-1'), three of the better inhibitors were tested at higher levels of diastereomeric purity. The diastereomers corresponding to the active forms of the 1-(ethyl)propylamide, the diethylamide, and the dipropylamide (all of the same configuration at the C-1' chiral center), and the diastereomer of the diethylamide representing the active form at both C-1' and C-1 were isolated or synthesized from precursors that were isolated as diastereomers. These diastereomers showed some improvement in neuraminidase inhibition over the parent diastereomeric mixtures. 1-Carboxy-1-hydroxy derivatives of the best active compounds, the diethylamide and the dipropylamide, were also prepared. These compounds were not as active as the compounds without the 1-hydroxy group. In an in vivo study, the C-1' active isomer of the diethylamide from the 1-carboxy series was tested in influenza-infected mice by oral and intranasal administration and found to be very effective only intranasally in preventing weight loss at doses as low as 0

  18. Aspects of gene structure and functional regulation of the isozymes of Na,K-ATPase

    DEFF Research Database (Denmark)

    Jorgensen, P.L.

    2001-01-01

    Gaps in our understanding of the complex regulated expression of isozymes of Na,K-ATPase and the diverse systems for posttranslational modification and short term regulation of active Na,K-transport in animals and humans are the main problems in comprehensive Na,K-pump physiology. In mammalian...... genomes, the genes of four alpha-subunit and at least three beta-subunit isoforms of Na,K-ATPase are identified and two gamma-subunits are expressed in kidney. The isoforms combine in a number of Na,K-ATPase isozymes that are expressed in a tissue and cell specific manner. Models of the molecular...... mechanism of regulation of these isozymes have become more reliable due to progress in understanding the three-dimensional protein structure and conformational transitions mediating transfer of energy from the P-domain to intramembrane Na+ and K+ binding sites....

  19. A generic system for the expression and purification of soluble and stable influenza neuraminidase.

    Directory of Open Access Journals (Sweden)

    Peter M Schmidt

    Full Text Available The influenza surface glycoprotein neuraminidase (NA is essential for the efficient spread of the virus. Antiviral drugs such as Tamiflu (oseltamivir and Relenza (zanamivir that inhibit NA enzyme activity have been shown to be effective in the treatment of influenza infections. The recent 'swine flu' pandemic and world-wide emergence of Tamiflu-resistant seasonal human influenza A(H1N1 H(274Y have highlighted the need for the ongoing development of new anti-virals, efficient production of vaccine proteins and novel diagnostic tools. Each of these goals could benefit from the production of large quantities of highly pure and stable NA. This publication describes a generic expression system for NAs in a baculovirus Expression Vector System (BEVS that is capable of expressing milligram amounts of recombinant NA. To construct NAs with increased stability, the natural influenza NA stalk was replaced by two different artificial tetramerization domains that drive the formation of catalytically active NA homotetramers: GCN4-pLI from yeast or the Tetrabrachion tetramerization domain from Staphylothermus marinus. Both recombinant NAs are secreted as FLAG-tagged proteins to allow for rapid and simple purification. The Tetrabrachion-based NA showed good solubility, increased stability and biochemical properties closer to the original viral NA than the GCN4-pLI based construct. The expressed quantities and high quality of the purified recombinant NA suggest that this expression system is capable of producing recombinant NA for a broad range of applications including high-throughput drug screening, protein crystallisation, or vaccine development.

  20. Composition of hemagglutinin and neuraminidase affects the antigen yield of influenza A(H1N1)pdm09 candidate vaccine viruses.

    Science.gov (United States)

    Shirakura, Masayuki; Kawaguchi, Akira; Tashiro, Masato; Nobusawa, Eri

    2013-01-01

    To improve the hemagglutinin (HA) antigen yield of influenza A(H1N1)pdm09 candidate vaccine viruses, we generated 7:1, 6:2, and 5:3 genetic reassortant viruses between wild-type (H1N1)pdm09 (A/California/7/2009) (Cal7) and a high-yielding master virus, A/Puerto Rico/8/34 (PR8). These viruses contained the HA; HA and neuraminidase (NA); and HA, NA, and M genes, respectively, derived from Cal7, on a PR8 backbone. The influence of the amino acid residue at position 223 in Cal7 HA on virus growth and HA antigen yield differed between these reassortant viruses. NIIDRG-7, a 7:1 virus possessing arginine at position 223, exhibited a 10-fold higher 50% egg infectious dose (EID(50)) (10.0 log(10)EID(50)/ml) than the 5:3 and 6:2 viruses. It also had 1.5- to 3-fold higher protein (13.8 μg/ml of allantoic fluids) and HA antigen (4.1 μg/ml of allantoic fluids) yields than the 5:3 and 6:2 viruses, which possessed identical Cal7 HA proteins. However, the HA antigen yield of the other 7:1 virus, which possessed glutamine at position 223 was 60% of that of NIIDRG-7. In addition, a novel 6:2 virus possessing Cal7 HA and the NA of A/Wisconsin/10/98 (a triple reassortant swine-like H1N1 virus), produced 107% of the HA yield of NIIDRG-7. In this study, we showed that the balance between HA and NA in the influenza A(H1N1)pdm09 virus affects its protein and antigen yield.

  1. Structure-function analysis of two variants of mumps virus hemagglutinin-neuraminidase protein

    Directory of Open Access Journals (Sweden)

    Gerardo Santos-López

    Full Text Available A point mutation from guanine (G to adenine (A at nucleotide position 1081 in the hemagglutinin-neuraminidase (HN gene has been associated with neurovirulence of Urabe AM9 mumps virus vaccine. This mutation corresponds to a glutamic acid (E to lysine (K change at position 335 in the HN glycoprotein. We have experimentally demonstrated that two variants of Urabe AM9 strain (HN-A1081 and HN-G1081 differ in neurotropism, sialic acidbinding affinity and neuraminidase activity. In the present study, we performed a structure-function analysis of that amino acid substitution; the structures of HN protein of both Urabe AM9 strain variants were predicted. Based on our analysis, the E/K mutation changes the protein surface properties and to a lesser extent their conformations, which in turn reflects in activity changes. Our modeling results suggest that this E/K interchange does not affect the structure of the sialic acid binding motif; however, the electrostatic surface differs drastically due to an exposed short alpha helix. Consequently, this mutation may affect the accessibility of HN to substrates and membrane receptors of the host cells. Our findings appear to explain the observed differences in neurotropism of these vaccine strains.

  2. Neuraminidase-mediated, NKp46-dependent immune-evasion mechanism of influenza viruses.

    Science.gov (United States)

    Bar-On, Yotam; Glasner, Ariella; Meningher, Tal; Achdout, Hagit; Gur, Chamutal; Lankry, Dikla; Vitenshtein, Alon; Meyers, Adrienne F A; Mandelboim, Michal; Mandelboim, Ofer

    2013-04-25

    Natural killer (NK) cells play an essential role in the defense against influenza virus, one of the deadliest respiratory viruses known today. The NKp46 receptor, expressed by NK cells, is critical for controlling influenza infections, as influenza-virus-infected cells are eliminated through the recognition of the viral hemagglutinin (HA) protein by NKp46. Here, we describe an immune-evasion mechanism of influenza viruses that is mediated by the neuraminidase (NA) protein. By using various NA blockers, we show that NA removes sialic acid residues from NKp46 and that this leads to reduced recognition of HA. Furthermore, we provide in vivo and in vitro evidence for the existence of this NA-mediated, NKp46-dependent immune-evasion mechanism and demonstrate that NA inhibitors, which are commonly used for the treatment of influenza infections, are useful not only as blockers of virus budding but also as boosters of NKp46 recognition. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

  3. Synthesis of Sulfo-Sialic Acid Analogues: Potent Neuraminidase Inhibitors in Regards to Anomeric Functionality.

    Science.gov (United States)

    Vavricka, Christopher J; Muto, Chiaki; Hasunuma, Tomohisa; Kimura, Yoshinobu; Araki, Michihiro; Wu, Yan; Gao, George F; Ohrui, Hiroshi; Izumi, Minoru; Kiyota, Hiromasa

    2017-08-15

    The design, synthesis and application of N-acetylneuraminic acid-derived compounds bearing anomeric sulfo functional groups are described. These novel compounds, which we refer to as sulfo-sialic acid analogues, include 2-decarboxy-2-deoxy-2-sulfo-N-acetylneuraminic acid and its 4-deoxy-3,4-dehydrogenated pseudoglycal. While 2-decarboxy-2-deoxy-2-sulfo-N-acetylneuraminic acid contains no further modifications of the 2-deoxy-pyranose ring, it is still a more potent inhibitor of avian-origin H5N1 neuraminidase (NA) and drug-resistant His275Tyr NA as compared to the oxocarbenium ion transition state analogue 2,3-dehydro-2-deoxy-N-acetylneuraminic acid. The sulfo-sialic acid analogues described in this report are also more potent inhibitors of influenza NA (up to 40-fold) and bacterial NA (up to 8.5-fold) relative to the corresponding anomeric phosphonic acids. These results confirm that this novel anomeric sulfo modification offers great potential to improve the potency of next-generation NA inhibitors including covalent inhibitors.

  4. Design, in silico studies, synthesis and in vitro evaluation of oseltamivir derivatives as inhibitors of neuraminidase from influenza A virus H1N1.

    Science.gov (United States)

    Neri-Bazán, Rocío M; García-Machorro, Jazmín; Méndez-Luna, David; Tolentino-López, Luis E; Martínez-Ramos, Federico; Padilla-Martínez, Itzia I; Aguilar-Faisal, Leopoldo; Soriano-Ursúa, Marvin A; Trujillo-Ferrara, José G; Fragoso-Vázquez, M Jonathan; Barrón, Blanca L; Correa-Basurto, José

    2017-03-10

    Since the neuraminidase (NA) enzyme of the influenza A virus plays a key role in the process of release of new viral particles from a host cell, it is often a target for new drug design. The emergence of NA mutations, such as H275Y, has led to great resistance against neuraminidase inhibitors, including oseltamivir and zanamivir. Hence, we herein designed a set of derivatives by modifying the amine and/or carboxylic groups of oseltamivir. After being screened for their physicochemical (Lipinski's rule) and toxicological properties, the remaining compounds were submitted to molecular and theoretical studies. The docking simulations provided insights into NA recognition patterns, demonstrating that oseltamivir modified at the carboxylic moiety and coupled with anilines had higher affinity and a better binding pose for NA than the derivatives modified at the amine group. Based on these theoretical studies, the new oseltamivir derivatives may have higher affinity to mutant variants and possibly to other viral subtypes. Accordingly, two compounds were selected for synthesis, which together with their respective intermediates were evaluated for their cytotoxicity and antiviral activities. Their biological activity was then tested in cells infected with the A/Puerto Rico/916/34 (H1N1) influenza virus, and virus yield reduction assays were performed. Additionally, by measuring neuraminidase activity with the neuraminidase assay kit it was found that the compounds produced inhibitory activity on this enzyme. Finally, the infected cells were analysed with atomic force microscopy (AFM), observing morphological changes strongly suggesting that these compounds interfered with cellular release of viral particles. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  5. An Na+/H+ antiporter gene from wheat plays an important role in ...

    Indian Academy of Sciences (India)

    A vacuole Na+/H+ antiporter gene TaNHX2 was obtained by screening the wheat cDNA library and by the 5′-RACE method. The expression of TaNHX2 was induced in roots and leaves by treatment with NaCl, polyethylene glycol (PEG), cold and abscisic acid (ABA). When expressed in a yeast mutant (nhx1), TaNHX2 ...

  6. Positive regulation of insulin signaling by neuraminidase 1.

    Science.gov (United States)

    Dridi, Larbi; Seyrantepe, Volkan; Fougerat, Anne; Pan, Xuefang; Bonneil, Eric; Thibault, Pierre; Moreau, Allain; Mitchell, Grant A; Heveker, Nikolaus; Cairo, Christopher W; Issad, Tarik; Hinek, Alexander; Pshezhetsky, Alexey V

    2013-07-01

    Neuraminidases (sialidases) catalyze the removal of sialic acid residues from sialylated glycoconjugates. We now report that mammalian neuraminidase 1 (Neu1), in addition to its catabolic function in lysosomes, is transported to the cell surface where it is involved in the regulation of insulin signaling. Insulin binding to its receptor rapidly induces interaction of the receptor with Neu1, which hydrolyzes sialic acid residues in the glycan chains of the receptor and, consequently, induces its activation. Cells from sialidosis patients with a genetic deficiency of Neu1 show impairment of insulin-induced phosphorylation of downstream protein kinase AKT, and treatment of these cells with purified Neu1 restores signaling. Genetically modified mice with ∼10% of the normal Neu1 activity exposed to a high-fat diet develop hyperglycemia and insulin resistance twice as fast as their wild-type counterparts. Together, these studies identify Neu1 as a novel component of the signaling pathways of energy metabolism and glucose uptake.

  7. Exploring the chemical space of influenza neuraminidase inhibitors

    Directory of Open Access Journals (Sweden)

    Nuttapat Anuwongcharoen

    2016-04-01

    Full Text Available The fight against the emergence of mutant influenza strains has led to the screening of an increasing number of compounds for inhibitory activity against influenza neuraminidase. This study explores the chemical space of neuraminidase inhibitors (NAIs, which provides an opportunity to obtain further molecular insights regarding the underlying basis of their bioactivity. In particular, a large set of 347 and 175 NAIs against influenza A and B, respectively, was compiled from the literature. Molecular and quantum chemical descriptors were obtained from low-energy conformational structures geometrically optimized at the PM6 level. The bioactivities of NAIs were classified as active or inactive according to their half maximum inhibitory concentration (IC50 value in which IC50 < 1µM and ≥ 10µM were defined as active and inactive compounds, respectively. Interpretable decision rules were derived from a quantitative structure–activity relationship (QSAR model established using a set of substructure descriptors via decision tree analysis. Univariate analysis, feature importance analysis from decision tree modeling and molecular scaffold analysis were performed on both data sets for discriminating important structural features amongst active and inactive NAIs. Good predictive performance was achieved as deduced from accuracy and Matthews correlation coefficient values in excess of 81% and 0.58, respectively, for both influenza A and B NAIs. Furthermore, molecular docking was employed to investigate the binding modes and their moiety preferences of active NAIs against both influenza A and B neuraminidases. Moreover, novel NAIs with robust binding fitness towards influenza A and B neuraminidase were generated via combinatorial library enumeration and their binding fitness was on par or better than FDA-approved drugs. The results from this study are anticipated to be beneficial for guiding the rational drug design of novel NAIs for treating influenza

  8. Computational analysis and determination of a highly conserved surface exposed segment in H5N1 avian flu and H1N1 swine flu neuraminidase

    Directory of Open Access Journals (Sweden)

    Nandy Ashesh

    2010-02-01

    Full Text Available Abstract Background Catalytic activity of influenza neuraminidase (NA facilitates elution of progeny virions from infected cells and prevents their self-aggregation mediated by the catalytic site located in the body region. Research on the active site of the molecule has led to development of effective inhibitors like oseltamivir, zanamivir etc, but the high rate of mutation and interspecies reassortment in viral sequences and the recent reports of oseltamivir resistant strains underlines the importance of determining additional target sites for developing future antiviral compounds. In a recent computational study of 173 H5N1 NA gene sequences we had identified a 50-base highly conserved region in 3'-terminal end of the NA gene. Results We extend the graphical and numerical analyses to a larger number of H5N1 NA sequences (514 and H1N1 swine flu sequences (425 accessed from GenBank. We use a 2D graphical representation model for the gene sequences and a Graphical Sliding Window Method (GSWM for protein sequences scanning the sequences as a block of 16 amino acids at a time. Using a protein sequence descriptor defined in our model, the protein sliding scan method allowed us to compare the different strains for block level variability, which showed significant statistical correlation to average solvent accessibility of the residue blocks; single amino acid position variability results in no correlation, indicating the impact of stretch variability in chemical environment. Close to the C-terminal end the GSWM showed less descriptor-variability with increased average solvent accessibility (ASA that is also supported by conserved predicted secondary structure of 3' terminal RNA and visual evidence from 3D crystallographic structure. Conclusion The identified terminal segment, strongly conserved in both RNA and protein sequences, is especially significant as it is surface exposed and structural chemistry reveals the probable role of this stretch in

  9. Neuraminidase as an enzymatic marker for detecting airborne Influenza virus and other viruses.

    Science.gov (United States)

    Turgeon, Nathalie; Toulouse, Marie-Josée; Ho, Jim; Li, Dongqing; Duchaine, Caroline

    2017-02-01

    Little information is available regarding the effectiveness of air samplers to collect viruses and regarding the effects of sampling processes on viral integrity. The neuraminidase enzyme is present on the surface of viruses that are of agricultural and medical importance. It has been demonstrated that viruses carrying this enzyme can be detected using commercial substrates without having to process the sample by methods such as RNA extraction. This project aims at evaluating the effects of 3 aerosol-sampling devices on the neuraminidase enzyme activity of airborne viruses. The purified neuraminidase enzymes from Clostridium perfringens, a strain of Influenza A (H1N1) virus, the FluMist influenza vaccine, and the Newcastle disease virus were used as models. The neuraminidase models were aerosolized in aerosol chambers and sampled with 3 different air samplers (SKC BioSampler, 3-piece cassettes with polycarbonate filters, and Coriolis μ) to assess the effect on neuraminidase enzyme activity. Our results demonstrated that Influenza virus and Newcastle disease virus neuraminidase enzymes are resistant to aerosolization and sampling with all air samplers tested. Moreover, we demonstrated that the enzymatic neuraminidase assay is as sensitive as RT-qPCR for detecting low concentrations of Influenza virus and Newcastle disease virus. Therefore, given the sensitivity of the assay and its compatibility with air sampling methods, viruses carrying the neuraminidase enzyme can be rapidly detected from air samples using neuraminidase activity assay without having to preprocess the samples.

  10. Structural Characterization of the 1918 Influenza H1N1 Neuraminidase

    Energy Technology Data Exchange (ETDEWEB)

    Xu, X.; Zhu, X.; Dwek, R.A.; Stevens, J.; Wilson, I.A.

    2009-05-28

    Influenza virus neuraminidase (NA) plays a crucial role in facilitating the spread of newly synthesized virus in the host and is an important target for controlling disease progression. The NA crystal structure from the 1918 'Spanish flu' (A/Brevig Mission/1/18 H1N1) and that of its complex with zanamivir (Relenza) at 1.65-{angstrom} and 1.45-{angstrom} resolutions, respectively, corroborated the successful expression of correctly folded NA tetramers in a baculovirus expression system. An additional cavity adjacent to the substrate-binding site is observed in N1, compared to N2 and N9 NAs, including H5N1. This cavity arises from an open conformation of the 150 loop (Gly147 to Asp151) and appears to be conserved among group 1 NAs (N1, N4, N5, and N8). It closes upon zanamivir binding. Three calcium sites were identified, including a novel site that may be conserved in N1 and N4. Thus, these high-resolution structures, combined with our recombinant expression system, provide new opportunities to augment the limited arsenal of therapeutics against influenza.

  11. Peramivir susceptibilities of recombinant influenza A and B variants selected with various neuraminidase inhibitors.

    Science.gov (United States)

    Fage, Clément; Tu, Véronique; Carbonneau, Julie; Abed, Yacine; Boivin, Guy

    2017-03-22

    Peramivir is a parenteral neuraminidase inhibitor (NAI) approved for treating influenza infections in a few countries. We determined peramivir susceptibilities of several uncharacterized influenza A and B neuraminidase (NA) and haemagglutinin (HA) mutants selected with different NAIs. Recombinant wild-type (WT) and mutant NA proteins were expressed in 293T cells and susceptibility to peramivir, oseltamivir and zanamivir was determined by NA inhibition assay using the MUNANA substrate. Recombinant/reassortant influenza A(H1N1), A(H3N2) and B HA mutants were rescued by reverse genetics and assessed by plaque size or viral yield assays for drug susceptibility. Recombinant R152K, I222K/T, G248R+I266V, Q312R+I427T and R371K (A[H1N1]pdm09); E41G, 1222L/V, Q226H and S247P (A[H3N2]) and D198Y, A246D/S/T and G402S (B) mutant NA proteins (N2 numbering) were analysed. Peramivir exhibited the lowest IC50 values against both influenza A and B WT NAs. Peramivir and oseltamivir generally shared similar phenotypes. Of note, peramivir retained activity against I222K/T (A[H1N1]pdm09), I222L/V (A[H3N2]) and A246T (B) mutants, which had reduced inhibition (RI) or highly RI (HRI) against oseltamivir. Cross-RI/HRI against the three NAIs was observed for R152K, R371K and Q312R+I427T (A[H1N1]pdm09); S247P (A[H3N2]) and D198Y (B) mutants. All tested recombinant/reassortant R208K (A/Puerto Rico/8/34 [H1N1]); A28T, R124M and K189E (A/Victoria/3/75 [H3N2]) and T139N (B/Phuket/3073/13) HA mutants were susceptible to peramivir in cell culture experiments. Peramivir is highly active against seasonal influenza subtypes. Although peramivir and oseltamivir generally share similar phenotypes, peramivir still possesses activity against some variants with RI/HRI against oseltamivir. Finally, NAI-induced HA substitutions alone did not significantly impact NAI susceptibility.

  12. Analise molecular do gene RHCE e suas variantes na população brasileira

    OpenAIRE

    Karina Antero Ribeiro Rosa

    2005-01-01

    Resumo: O sistema Rh representa atualmente um dos sistemas de grupos sangüíneos de maior interesse devido a sua importância na clínica transfusional e na medicina materno-feta!. Os antígenos do sistema Rh são codificados por dois genes altamente homólogos, RHD que produz o antígeno D e o gene RHCE (alelos RH Ce, RH cE, RH ce e RH CE), que produz dois pares de antígenos antitéticos, C/c e Ele. Variantes que apresentam expressão parcial ou ausência total dos produtos do gene RHCE são relativame...

  13. Cloning and characterization of a novel vacuolar Na+/H+ antiporter gene (Dgnhx1 from chrysanthemum.

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    Qing-Lin Liu

    Full Text Available Plant vacuolar Na(+/H(+ antiporter genes play significant roles in salt tolerance. However, the roles of the chrysanthemum vacuolar Na(+/H(+ antiporter genes in salt stress response remain obscure. In this study, we isolated and characterized a novel vacuolar Na(+/H(+ antiporter gene DgNHX1 from chrysanthemum. The DgNHX1 sequence contained 1920 bp with a complete open reading frame of 1533 bp encoding a putative protein of 510 amino acids with a predicted protein molecular weight of 56.3 kDa. DgNHX1 was predicted containing nine transmembrane domains. Its expression in the chrysanthemum was up-regulated by salt stress, but not by abscisic acid (ABA. To assess roles of DgNHX1 in plant salt stress responses, we performed gain-of-function experiment. The DgNHX1-overexpression tobacco plants showed significant salt tolerance than the wild type (WT. The transgenic lines exhibited more accumulation of Na(+ and K(+ under salt stress. These findings suggest that DgNHX1 plays a positive regulatory role in salt stress response.

  14. Neuraminidase stalk length and additional glycosylation of the hemagglutinin influence the virulence of influenza H5N1 viruses for mice.

    Science.gov (United States)

    Matsuoka, Yumiko; Swayne, David E; Thomas, Colleen; Rameix-Welti, Marie-Anne; Naffakh, Nadia; Warnes, Christine; Altholtz, Melanie; Donis, Ruben; Subbarao, Kanta

    2009-05-01

    Following circulation of avian influenza H5 and H7 viruses in poultry, the hemagglutinin (HA) can acquire additional glycosylation sites, and the neuraminidase (NA) stalk becomes shorter. We investigated whether these features play a role in the pathogenesis of infection in mammalian hosts. From 1996 to 2007, H5N1 viruses with a short NA stalk have become widespread in several avian species. Compared to viruses with a long-stalk NA, viruses with a short-stalk NA showed a decreased capacity to elute from red blood cells and an increased virulence in mice, but not in chickens. The presence of additional HA glycosylation sites had less of an effect on virulence than did NA stalk length. The short-stalk NA of H5N1 viruses circulating in Asia may contribute to virulence in humans.

  15. Pitfalls in Diagnosing Neuraminidase Deficiency : Psychosomatics and Normal Sialic Acid Excretion

    NARCIS (Netherlands)

    Schene, Imre F; Ayuso, Viera Kalinina|info:eu-repo/dai/nl/33793259X; de Sain-van der Velden, Monique|info:eu-repo/dai/nl/304817910; van Gassen, Koen L I|info:eu-repo/dai/nl/304819417; Cuppen, Inge|info:eu-repo/dai/nl/314436529; van Hasselt, Peter M|info:eu-repo/dai/nl/304814423; Visser, Gepke|info:eu-repo/dai/nl/229317057

    2015-01-01

    Neuraminidase deficiency (mucolipidosis I, sialidosis types I and II, cherry-red spot myoclonus syndrome) is a lysosomal storage disorder with an expanding clinical phenotype. Here, we report the striking diagnostic history of late-onset neuraminidase deficiency in two sisters, currently aged 14

  16. ESTIMATION OF Na GENE FREQUENCY ON NATIVE CHICKEN POPULATION AND ITS EFFECT ON HATCHABILITY PERFORMANCE

    Directory of Open Access Journals (Sweden)

    J. Setianto

    2014-10-01

    Full Text Available The objective of this research was to estimate the Na gene frequency of the native chickenpopulation and its effect on hatchability performance. The research used Native Chickens with Nakedneckphenotype and Normal-feathered Native Chicken, consisted of 16 cocks and 48 hens as the parent.They were put in 4 breeding groups : 1 Breeding group of Legund Cock with Legund Hen (LL; 2Legund Cock with Normal-feathered Native Hen (LN; 3 Normal-feathered Native Cock with LegundHen (NL; 4 Normal-feathered Native Cock with Normal-feathered Native Hen (NN. Each groupconsisted of 4 replications and each replication consisted of 1 Cock with 3 Hens. CompletelyRandomized Design (CRD were used to analysis the fertility, hatchability and embryo mortality.Hardy-Weinberg’s Law of Gene equilibrium method was used to find out the spreading estimation of Naand na gene in whole offspring population. Experimental result showed that the breeding groups had noinfluence (P>0.05 on egg fertility, hatchability and embryo mortality. The phenotypes of the offspringof the breeding groups were not deviate from the Mendel’s law. The estimation of the Na gene frequencywas 19.7%.

  17. Structural basis for substrate specificity of mammalian neuraminidases.

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    Victoria Smutova

    Full Text Available The removal of sialic acid (Sia residues from glycoconjugates in vertebrates is mediated by a family of neuraminidases (sialidases consisting of Neu1, Neu2, Neu3 and Neu4 enzymes. The enzymes play distinct physiological roles, but their ability to discriminate between the types of linkages connecting Sia and adjacent residues and between the identity and arrangement of the underlying sugars has never been systematically studied. Here we analyzed the specificity of neuraminidases by studying the kinetics of hydrolysis of BODIPY-labeled substrates containing common mammalian sialylated oligosaccharides: 3'Sia-LacNAc, 3'SiaLac, SiaLex, SiaLea, SiaLec, 6'SiaLac, and 6'SiaLacNAc. We found significant differences in substrate specificity of the enzymes towards the substrates containing α2,6-linked Sia, which were readily cleaved by Neu3 and Neu1 but not by Neu4 and Neu2. The presence of a branching 2-Fuc inhibited Neu2 and Neu4, but had almost no effect on Neu1 or Neu3. The nature of the sugar residue at the reducing end, either glucose (Glc or N-acetyl-D-glucosamine (GlcNAc had only a minor effect on all neuraminidases, whereas core structure (1,3 or 1,4 bond between D-galactose (Gal and GlcNAc was found to be important for Neu4 strongly preferring β3 (core 1 to β4 (core 2 isomer. Neu3 and Neu4 were in general more active than Neu1 and Neu2, likely due to their preference for hydrophobic substrates. Neu2 and Neu3 were examined by molecular dynamics to identify favorable substrate orientations in the binding sites and interpret the differences in their specificities. Finally, using knockout mouse models, we confirmed that the substrate specificities observed in vitro were recapitulated in enzymes found in mouse brain tissues. Our data for the first time provide evidence for the characteristic substrate preferences of neuraminidases and their ability to discriminate between distinct sialoside targets.

  18. Virtual screening approach to identifying influenza virus neuraminidase inhibitors using molecular docking combined with machine-learning-based scoring function.

    Science.gov (United States)

    Zhang, Li; Ai, Hai-Xin; Li, Shi-Meng; Qi, Meng-Yuan; Zhao, Jian; Zhao, Qi; Liu, Hong-Sheng

    2017-10-10

    In recent years, an epidemic of the highly pathogenic avian influenza H7N9 virus has persisted in China, with a high mortality rate. To develop novel anti-influenza therapies, we have constructed a machine-learning-based scoring function (RF-NA-Score) for the effective virtual screening of lead compounds targeting the viral neuraminidase (NA) protein. RF-NA-Score is more accurate than RF-Score, with a root-mean-square error of 1.46, Pearson's correlation coefficient of 0.707, and Spearman's rank correlation coefficient of 0.707 in a 5-fold cross-validation study. The performance of RF-NA-Score in a docking-based virtual screening of NA inhibitors was evaluated with a dataset containing 281 NA inhibitors and 322 noninhibitors. Compared with other docking-rescoring virtual screening strategies, rescoring with RF-NA-Score significantly improved the efficiency of virtual screening, and a strategy that averaged the scores given by RF-NA-Score, based on the binding conformations predicted with AutoDock, AutoDock Vina, and LeDock, was shown to be the best strategy. This strategy was then applied to the virtual screening of NA inhibitors in the SPECS database. The 100 selected compounds were tested in an in vitro H7N9 NA inhibition assay, and two compounds with novel scaffolds showed moderate inhibitory activities. These results indicate that RF-NA-Score improves the efficiency of virtual screening for NA inhibitors, and can be used successfully to identify new NA inhibitor scaffolds. Scoring functions specific for other drug targets could also be established with the same method.

  19. Isolation of Panels of Llama Single-Domain Antibody Fragments Binding All Nine Neuraminidase Subtypes of Influenza A Virus

    Directory of Open Access Journals (Sweden)

    Guus Koch

    2013-04-01

    Full Text Available Avian influenza A virus comprises sixteen hemagglutinin (HA and nine neuraminidase (NA subtypes (N1–N9. To isolate llama single-domain antibody fragments (VHHs against all N subtypes, four llamas were immunized with mixtures of influenza viruses. Selections using influenza virus yielded predominantly VHHs binding to the highly immunogenic HA and nucleoprotein. However, selection using enzymatically active recombinant NA (rNA protein enabled us to isolate NA binding VHHs. Some isolated VHHs cross-reacted to other N subtypes. These were subsequently used for the capture of N subtypes that could not be produced as recombinant protein (rN6 or were enzymatically inactive (rN1, rN5 in phage display selection, yielding novel VHHs. In total we isolated 188 NA binding VHHs, 64 of which were expressed in yeast. Most VHHs specifically recognize a single N subtype, but some VHHs cross-react with other N-subtypes. At least one VHH bound to all N subtypes, except N4, identifying a conserved antigenic site. Thus, this work (1 describes methods for isolating NA binding VHHs, (2 illustrates the suitability of llama immunization with multiple antigens for retrieving many binders against different antigens and (3 describes 64 novel NA binding VHHs, including a broadly reactive VHH, which can be used in various assays for influenza virus subtyping, detection or serology.

  20. [Study on gene expression of Tamarix under NaHCO3 stress using SSH technology].

    Science.gov (United States)

    Yang, Chuan-Ping; Wang, Yu-Cheng; Liu, Gui-Feng; Jiang, Jing

    2004-09-01

    The gene expression of Tamarix androssowii under NaHCO3 stresses is studied by using SSH technique, in which the cDNA from the materials treated with NaHCO3 solution is as tester and the cDNA from the materials in normal growth is as driver. Total 36 genes related to NaHCO3 stress were obtained through Northern hybridization. Blastx analysis showed that the proteins encoded by these genes were homologous to the following proteins: the antioxidant enzymes catalase and peroxiredoxin; trehalose phosphatase, which was related to trehalose synthesis; a few regulation proteins such as bZIP transcription factor, MADS-box protein, glycine-rich RNA-binding proteins, CCCH-type zinc finger protein and F-box protein etc; early light-induced protein, which could protect and/or repair the photosynthetic apparatus damage induced by stress; cysteine proteinase and vacuolar processing enzyme that can make function in plant cell death, and lipid transfer protein precursor, polyubiquitin, chalcone synthase, NADP-dependent isocitrate dehydrogenase, salt-induced S12 protein, and oxygen-evolving enhancer protein 1 etc. Among 36 genes obtained, the proteins encoded three genes were homologous to 3 putative proteins: HAK2, calcium-binding protein and RNA-binding protein, respectively. In addition, 6 new salt stress response squences were found. The result indicated that the salt-tolerant mechanism of Tamarix androssowii may be a complicated, interactive system involving multiple approaches and multiple genes, but not only a single salt gland-depended approach.

  1. Comprehensive transcriptional profiling of NaCl-stressed Arabidopsis roots reveals novel classes of responsive genes

    Directory of Open Access Journals (Sweden)

    Deyholos Michael K

    2006-10-01

    Full Text Available Abstract Background Roots are an attractive system for genomic and post-genomic studies of NaCl responses, due to their primary importance to agriculture, and because of their relative structural and biochemical simplicity. Excellent genomic resources have been established for the study of Arabidopsis roots, however, a comprehensive microarray analysis of the root transcriptome following NaCl exposure is required to further understand plant responses to abiotic stress and facilitate future, systems-based analyses of the underlying regulatory networks. Results We used microarrays of 70-mer oligonucleotide probes representing 23,686 Arabidopsis genes to identify root transcripts that changed in relative abundance following 6 h, 24 h, or 48 h of hydroponic exposure to 150 mM NaCl. Enrichment analysis identified groups of structurally or functionally related genes whose members were statistically over-represented among up- or down-regulated transcripts. Our results are consistent with generally observed stress response themes, and highlight potentially important roles for underappreciated gene families, including: several groups of transporters (e.g. MATE, LeOPT1-like; signalling molecules (e.g. PERK kinases, MLO-like receptors, carbohydrate active enzymes (e.g. XTH18, transcription factors (e.g. members of ZIM, WRKY, NAC, and other proteins (e.g. 4CL-like, COMT-like, LOB-Class 1. We verified the NaCl-inducible expression of selected transcription factors and other genes by qRT-PCR. Conclusion Micorarray profiling of NaCl-treated Arabidopsis roots revealed dynamic changes in transcript abundance for at least 20% of the genome, including hundreds of transcription factors, kinases/phosphatases, hormone-related genes, and effectors of homeostasis, all of which highlight the complexity of this stress response. Our identification of these transcriptional responses, and groups of evolutionarily related genes with either similar or divergent

  2. Structural analysis of the α subunit of Na(+)/K(+) ATPase genes in invertebrates.

    Science.gov (United States)

    Thabet, Rahma; Rouault, J-D; Ayadi, Habib; Leignel, Vincent

    2016-01-01

    The Na(+)/K(+) ATPase is a ubiquitous pump coordinating the transport of Na(+) and K(+) across the membrane of cells and its role is fundamental to cellular functions. It is heteromer in eukaryotes including two or three subunits (α, β and γ which is specific to the vertebrates). The catalytic functions of the enzyme have been attributed to the α subunit. Several complete α protein sequences are available, but only few gene structures were characterized. We identified the genomic sequences coding the α-subunit of the Na(+)/K(+) ATPase, from the whole-genome shotgun contigs (WGS), NCBI Genomes (chromosome), Genomic Survey Sequences (GSS) and High Throughput Genomic Sequences (HTGS) databases across distinct phyla. One copy of the α subunit gene was found in Annelida, Arthropoda, Cnidaria, Echinodermata, Hemichordata, Mollusca, Placozoa, Porifera, Platyhelminthes, Urochordata, but the nematodes seem to possess 2 to 4 copies. The number of introns varied from 0 (Platyhelminthes) to 26 (Porifera); and their localization and length are also highly variable. Molecular phylogenies (Maximum Likelihood and Maximum Parsimony methods) showed some clusters constituted by (Chordata/(Echinodermata/Hemichordata)) or (Plathelminthes/(Annelida/Mollusca)) and a basal position for Porifera. These structural analyses increase our knowledge about the evolutionary events of the α subunit genes in the invertebrates. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. Global update on the susceptibility of human influenza viruses to neuraminidase inhibitors, 2015-2016.

    Science.gov (United States)

    Gubareva, Larisa V; Besselaar, Terry G; Daniels, Rod S; Fry, Alicia; Gregory, Vicki; Huang, Weijuan; Hurt, Aeron C; Jorquera, Patricia A; Lackenby, Angie; Leang, Sook-Kwan; Lo, Janice; Pereyaslov, Dmitriy; Rebelo-de-Andrade, Helena; Siqueira, Marilda M; Takashita, Emi; Odagiri, Takato; Wang, Dayan; Zhang, Wenqing; Meijer, Adam

    2017-10-01

    Four World Health Organization (WHO) Collaborating Centres for Reference and Research on Influenza and one WHO Collaborating Centre for the Surveillance, Epidemiology and Control of Influenza (WHO CCs) assessed antiviral susceptibility of 14,330 influenza A and B viruses collected by WHO-recognized National Influenza Centres (NICs) between May 2015 and May 2016. Neuraminidase (NA) inhibition assay was used to determine 50% inhibitory concentration (IC50) data for NA inhibitors (NAIs) oseltamivir, zanamivir, peramivir and laninamivir. Furthermore, NA sequences from 13,484 influenza viruses were retrieved from public sequence databases and screened for amino acid substitutions (AAS) associated with reduced inhibition (RI) or highly reduced inhibition (HRI) by NAIs. Of the viruses tested by WHO CCs 93% were from three WHO regions: Western Pacific, the Americas and Europe. Approximately 0.8% (n = 113) exhibited either RI or HRI by at least one of four NAIs. As in previous seasons, the most common NA AAS was H275Y in A(H1N1)pdm09 viruses, which confers HRI by oseltamivir and peramivir. Two A(H1N1)pdm09 viruses carried a rare NA AAS, S247R, shown in this study to confer RI/HRI by the four NAIs. The overall frequency of A(H1N1)pdm09 viruses containing NA AAS associated with RI/HRI was approximately 1.8% (125/6915), which is slightly higher than in the previous 2014-15 season (0.5%). Three B/Victoria-lineage viruses contained a new AAS, NA H134N, which conferred HRI by zanamivir and laninamivir, and borderline HRI by peramivir. A single B/Victoria-lineage virus harboured NA G104E, which was associated with HRI by all four NAIs. The overall frequency of RI/HRI phenotype among type B viruses was approximately 0.6% (43/7677), which is lower than that in the previous season. Overall, the vast majority (>99%) of the viruses tested by WHO CCs were susceptible to all four NAIs, showing normal inhibition (NI). Hence, NAIs remain the recommended antivirals for treatment of

  4. Heterologous Expression of the Cotton NBS-LRR Gene GbaNA1 Enhances Verticillium Wilt Resistance in Arabidopsis

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    Nan-Yang Li

    2018-02-01

    Full Text Available Verticillium wilt caused by Verticillium dahliae results in severe losses in cotton, and is economically the most destructive disease of this crop. Improving genetic resistance is the cleanest and least expensive option to manage Verticillium wilt. Previously, we identified the island cotton NBS-LRR-encoding gene GbaNA1 that confers resistance to the highly virulent V. dahliae isolate Vd991. In this study, we expressed cotton GbaNA1 in the heterologous system of Arabidopsis thaliana and investigated the defense response mediated by GbaNA1 following inoculations with V. dahliae. Heterologous expression of GbaNA1 conferred Verticillium wilt resistance in A. thaliana. Moreover, overexpression of GbaNA1 enabled recovery of the resistance phenotype of A. thaliana mutants that had lost the function of GbaNA1 ortholog gene. Investigations of the defense response in A. thaliana showed that the reactive oxygen species (ROS production and the expression of genes associated with the ethylene signaling pathway were enhanced significantly following overexpression of GbaNA1. Intriguingly, overexpression of the GbaNA1 ortholog from Gossypium hirsutum (GhNA1 in A. thaliana did not induce the defense response of ROS production due to the premature termination of GhNA1, which lacks the encoded NB-ARC and LRR motifs. GbaNA1 therefore confers Verticillium wilt resistance in A. thaliana by the activation of ROS production and ethylene signaling. These results demonstrate the functional conservation of the NBS-LRR-encoding GbaNA1 in a heterologous system, and the mechanism of this resistance, both of which may prove valuable in incorporating GbaNA1-mediated resistance into other plant species.

  5. Expression profile of genes regulated by activity of the Na-H exchanger NHE1

    Directory of Open Access Journals (Sweden)

    Putney Luanna K

    2004-07-01

    Full Text Available Abstract Background In mammalian cells changes in intracellular pH (pHi, which are predominantly controlled by activity of plasma membrane ion exchangers, regulate a diverse range of normal and pathological cellular processes. How changes in pHi affect distinct cellular processes has primarily been determined by evaluating protein activities and we know little about how pHi regulates gene expression. Results A global profile of genes regulated in mammalian fibroblasts by decreased pHi induced by impaired activity of the plasma membrane Na-H exchanger NHE1 was characterized by using cDNA microarrays. Analysis of selected genes by quantitative RT-PCR, TaqMan, and immunoblot analyses confirmed results obtained from cDNA arrays. Consistent with established roles of pHi and NHE1 activity in cell proliferation and oncogenic transformation, grouping regulated genes into functional categories and biological pathways indicated a predominant number of genes with altered expression were associated with growth factor signaling, oncogenesis, and cell cycle progression. Conclusion A comprehensive analysis of genes selectively regulated by pHi provides insight on candidate targets that might mediate established effects of pHi on a number of normal and pathological cell functions.

  6. Genetic drift of HA and NA in Danish swine influenza virus from the period 2003-2012

    DEFF Research Database (Denmark)

    Fobian, Kristina; Breum, Solvej Østergaard; Hjulsager, Charlotte Kristiane

    2012-01-01

    The aim of this study is to analyze; the genetic drift in hemagglutinin (HA) and neuraminidase (NA) genes from influenza viruses isolated from Danish swine over the past decade; the antigenic evolution and relatedness between swine influenza virus strains of the H1 subtype by antigenic cartograph...... and along with the monitoring of antigenic changes in hemagglutinin subtypes it will be possible to ensure a continuous efficacy of influenza virus vaccines.......The aim of this study is to analyze; the genetic drift in hemagglutinin (HA) and neuraminidase (NA) genes from influenza viruses isolated from Danish swine over the past decade; the antigenic evolution and relatedness between swine influenza virus strains of the H1 subtype by antigenic cartography....... Currently at least three influenza A subtypes (H1N1, H1N2 and H3N2) are endemic in the Danish swine population, and since 2010 the pandemic virus (H1N1pdm09) have also frequently been detected. The focus in this study will be on H1N1 and H1N2, since the prevalence of H3N2 have declined over the past years...

  7. Different Origins of Newcastle Disease Virus Hemagglutinin-Neuraminidase Protein Modulate the Replication Efficiency and Pathogenicity of the Virus

    Directory of Open Access Journals (Sweden)

    Ji-hui Jin

    2017-08-01

    Full Text Available To investigate the exact effects of different origins of Newcastle disease virus (NDV hemagglutinin-neuraminidase (HN protein to the biological characteristics of the virus, we systematically studied the correlation between the HN protein and NDV virulence by exchanging the HN of velogenic or lentogenic NDV strains with the HN from other strains of different virulence. The results revealed that the rSG10 or rLaSota derivatives bearing the HN gene of other viruses exhibited decreased or increased hemadsorption (HAd, neuraminidase and fusion promotion activities. In vitro and in vivo tests further showed that changes in replication level, tissue tropism and virulence of the chimeric viruses were also consistent with these biological activities. These findings demonstrated that the balance among three biological activities caused variation in replication and pathogenicity of the virus, which was closely related to the origin of the HN protein. Our study highlights the importance of the HN glycoprotein in modulating the virulence of NDV and contributes to a more complete understanding of the virulence of NDV.

  8. Neuraminidase 1 is a Negative Regulator of Lysosomal Exocytosis

    Science.gov (United States)

    Yogalingam, Gouri; Bonten, Erik J.; van de Vlekkert, Diantha; Hu, Huimin; Moshiach, Simon; Connell, Samuel A.; d’Azzo, Alessandra

    2009-01-01

    SUMMARY Lysosomal exocytosis is a Ca2+-regulated mechanism that involves proteins responsible for cytoskeletal attachment and fusion of lysosomes with the plasma membrane. However, whether luminal lysosomal enzymes contribute to this process remains unknown. Here we show that neuraminidase Neu1 negatively regulates lysosomal exocytosis in hematopoietic cells by processing the sialic acids on the lysosomal membrane protein Lamp-1. In macrophages from Neu1-deficient mice, a model of the disease sialidosis, and in patients’ fibroblasts, oversialylated Lamp-1 enhances lysosomal exocytosis. Silencing of Lamp-1 reverts this phenotype by interfering with the docking of lysosomes at the plasma membrane. In Neu1-/- mice the excessive exocytosis of serine proteases in the bone niche leads to inactivation of extracellular serpins, premature degradation of VCAM-1, and loss of bone marrow retention. Our findings uncover an unexpected mechanism influencing lysosomal exocytosis and argue that exacerbations of this process form the basis for certain genetic diseases. PMID:18606142

  9. Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase.

    Science.gov (United States)

    Wu, Zhengliang L; Zhou, Hui; Ethen, Cheryl M; N Reinhold, Vernon

    2016-04-29

    The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. Yet to date, the structure component responsible for its virulence is still a mystery. In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. In this report, the N-glycans from both forms were released and characterized by mass spectrometry. It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. These findings were further supported by the results obtained from in vitro incorporation of azido fucose and (3)H-labeled fucose using core-6 fucosyltransferase, FUT8. In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  10. Influenza virus uses its neuraminidase protein to evade the recognition of two activating NK cell receptors.

    Science.gov (United States)

    Bar-On, Yotam; Seidel, Einat; Tsukerman, Pinchas; Mandelboim, Michal; Mandelboim, Ofer

    2014-08-01

    Natural Killer (NK) cells play a central role in the defense against viral infections and in the elimination of transformed cells. The recognition of pathogen-infected and tumor cells is controlled by inhibitory and activating receptors. We have previously shown that among the activating (killer) NK cell receptors the natural cytotoxicity receptors, NKp44 and NKp46, interact with the viral hemagglutinin (HA) protein expressed on the cell surface of influenza-virus-infected cells. We further showed that the interaction between NKp44/NKp46 and viral HA is sialic-acid dependent and that the recognition of HA by NKp44 and NKp46 leads to the elimination of the infected cells. Here we demonstrate that the influenza virus developed a counter-attack mechanism and that the virus uses its neuraminidase (NA) protein to prevent the recognition of HA by both the NKp44 and NKp46 receptors, resulting in reduced elimination of the infected cells by NK cells. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  11. Synergistic Antiviral Activity of S-033188/S-033447, a Novel Inhibitor of Influenza Virus Cap-Dependent Endonuclease, in Combination with Neuraminidase Inhibitors In Vitro

    OpenAIRE

    Kitano, Mitsutaka; Yamamoto, Atsuko; Noshi, Takeshi; Kawai, Makoto; Yoshida, Ryu; Sato, Akihiko; Shishido, Takao; Naito, Akira

    2017-01-01

    Abstract Background S-033447, an active form of orally available prodrug S-033188, is a novel small molecule inhibitor of cap-dependent endonuclease that is essential for influenza virus transcription and replication. In this study, we evaluated the inhibitory effect of S-033188 in combination with neuraminidase inhibitors on the replication of influenza A/H1N1 virus in cultured cells. Methods The inhibitory effects of S-033447 in combination with NA inhibitors on the cytopathic effect of A/P...

  12. Neuraminidase production by a Streptococcus sanguis strain associated with subacute bacterial endocarditis.

    OpenAIRE

    Straus, D. C.; Portnoy-Duran, C

    1983-01-01

    The properties of an extracellular neuraminidase produced by a Streptococcus sanguis strain (isolated from a confirmed case of subacute bacterial endocarditis) during growth in a defined medium was examined in this investigation. This enzyme, isolated from concentrated culture supernatants of S. sanguis biotype II, was active against human alpha-1 acid glycoprotein, N-acetylneuramin lactose, bovine submaxillary mucin, and fetuin. Neuraminidase production paralleled bacterial growth in defined...

  13. Parallel screening of wild-type and drug-resistant targets for anti-resistance neuraminidase inhibitors.

    Directory of Open Access Journals (Sweden)

    Kai-Cheng Hsu

    Full Text Available Infection with influenza virus is a major public health problem, causing serious illness and death each year. Emergence of drug-resistant influenza virus strains limits the effectiveness of drug treatment. Importantly, a dual H275Y/I223R mutation detected in the pandemic influenza A 2009 virus strain results in multidrug resistance to current neuraminidase (NA drugs. Therefore, discovery of new agents for treating multiple drug-resistant (MDR influenza virus infections is important. Here, we propose a parallel screening strategy that simultaneously screens wild-type (WT and MDR NAs, and identifies inhibitors matching the subsite characteristics of both NA-binding sites. These may maintain their potency when drug-resistant mutations arise. Initially, we analyzed the subsite of the dual H275Y/I223R NA mutant. Analysis of the site-moiety maps of NA protein structures show that the mutant subsite has a relatively small volume and is highly polar compared with the WT subsite. Moreover, the mutant subsite has a high preference for forming hydrogen-bonding interactions with polar moieties. These changes may drive multidrug resistance. Using this strategy, we identified a new inhibitor, Remazol Brilliant Blue R (RB19, an anthraquinone dye, which inhibited WT NA and MDR NA with IC(50 values of 3.4 and 4.5 µM, respectively. RB19 comprises a rigid core scaffold and a flexible chain with a large polar moiety. The former interacts with highly conserved residues, decreasing the probability of resistance. The latter forms van der Waals contacts with the WT subsite and yields hydrogen bonds with the mutant subsite by switching the orientation of its flexible side chain. Both scaffolds of RB19 are good starting points for lead optimization. The results reveal a parallel screening strategy for identifying resistance mechanisms and discovering anti-resistance neuraminidase inhibitors. We believe that this strategy may be applied to other diseases with high

  14. Neuraminidase activity of blue eye disease porcine rubulavirus: Specificity, affinity and inhibition studies.

    Science.gov (United States)

    Santos-López, Gerardo; Borraz-Argüello, María T; Márquez-Domínguez, Luis; Flores-Alonso, Juan Carlos; Ramírez-Mendoza, Humberto; Priem, Bernard; Fort, Sébastien; Vallejo-Ruiz, Verónica; Reyes-Leyva, Julio; Herrera-Camacho, Irma

    2017-10-01

    Porcine rubulavirus (PorPV), also known as La Piedad Michoacan Virus (LPMV) causes encephalitis and reproductive failure in newborn and adult pigs, respectively. The hemagglutinin-neuraminidase (HN) glycoprotein is the most exposed and antigenic of the virus proteins. HN plays central roles in PorPV infection; i.e., it recognizes sialic acid-containing cell receptors that mediate virus attachment and penetration; in addition, its neuraminidase (sialic acid releasing) activity has been proposed as a virulence factor. This work describes the purification and characterization of PorPV HN protein (isolate PAC1). The specificity of neuraminidase is restricted to sialyl(α2,3)lactose (3SL). HN showed typical Michaelis-Menten kinetics with fetuin as substrate (km=0.029μM, Vmax=522.8nmolmin-1mg-1). When 3SL was used as substrate, typical cooperative kinetics were found (S50=0.15μM, Vmax=154.3nmolmin-1mg-1). The influenza inhibitor zanamivir inhibited the PorPV neuraminidase with IC50 of 0.24μM. PorPV neuraminidase was activated by Ca2+ and inhibited by nucleoside triphosphates with the level of inhibition depending on phosphorylation level. The present results open possibilities to study the role of neuraminidase in the pathogenicity of PorPV infection and its potential inhibitors. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Robustification of Naïve Bayes Classifier and Its Application for Microarray Gene Expression Data Analysis

    OpenAIRE

    Ahmed, Md. Shakil; Shahjaman, Md.; Rana, Md. Masud; Mollah, Md. Nurul Haque

    2017-01-01

    The na?ve Bayes classifier (NBC) is one of the most popular classifiers for class prediction or pattern recognition from microarray gene expression data (MGED). However, it is very much sensitive to outliers with the classical estimates of the location and scale parameters. It is one of the most important drawbacks for gene expression data analysis by the classical NBC. The gene expression dataset is often contaminated by outliers due to several steps involved in the data generating process f...

  16. Vaccine-associated enhanced respiratory disease is influenced by haemagglutinin and neuraminidase in whole inactivated influenza virus vaccines.

    Science.gov (United States)

    Rajão, Daniela S; Chen, Hongjun; Perez, Daniel R; Sandbulte, Matthew R; Gauger, Phillip C; Loving, Crystal L; Shanks, G Dennis; Vincent, Amy

    2016-07-01

    Multiple subtypes and many antigenic variants of influenza A virus (IAV) co-circulate in swine in the USA, complicating effective use of commercial vaccines to control disease and transmission. Whole inactivated virus (WIV) vaccines may provide partial protection against IAV with substantial antigenic drift, but have been shown to induce vaccine-associated enhanced respiratory disease (VAERD) when challenged with an antigenic variant of the same haemagglutinin (HA) subtype. This study investigated the role the immune response against HA, neuraminidase (NA) and nucleoprotein (NP) may play in VAERD by reverse engineering vaccine and challenge viruses on a common backbone and using them in a series of vaccination/challenge trials. Mismatched HA between vaccine and challenge virus was necessary to induce VAERD. However, vaccines containing a matched NA abrogated the VAERD phenomenon induced by the HA mismatch and this was correlated with NA-inhibiting (NI) antibodies. Divergence between the two circulating swine N2 lineages (92 % identity) resulted in a loss of NI cross-reactivity and also resulted in VAERD with the mismatched HA. The NP lineage selected for use in the WIV vaccine strains did not affect protection or pathology. Thus the combination of HA and NA in the vaccine virus strains played a substantial role in vaccine protection versus immunopathology, suggesting that vaccines that target the HA protein alone could be more prone to VAERD due to the absence of cross-protective NI antibodies.

  17. Charged amino acid variability related to N-glyco -sylation and epitopes in A/H3N2 influenza: Hem -agglutinin and neuraminidase.

    Science.gov (United States)

    Huang, Zhong-Zhou; Yu, Liang; Huang, Ping; Liang, Li-Jun; Guo, Qing

    2017-01-01

    The A/H3N2 influenza viruses circulated in humans have been shown to undergo antigenic drift, a process in which amino acid mutations result from nucleotide substitutions. There are few reports regarding the charged amino acid mutations. The purpose of this paper is to explore the relations between charged amino acids, N-glycosylation and epitopes in hemagglutinin (HA) and neuraminidase (NA). A total of 700 HA genes (691 NA genes) of A/H3N2 viruses were chronologically analyzed for the mutational variants in amino acid features, N-glycosylation sites and epitopes since its emergence in 1968. It was found that both the number of HA N-glycosylation sites and the electric charge of HA increased gradually up to 2016. The charges of HA and HA1 increased respectively 1.54-fold (+7.0 /+17.8) and 1.08-fold (+8.0/+16.6) and the number of NGS in nearly doubled (7/12). As great diversities occurred in 1990s, involving Epitope A, B and D mutations, the charged amino acids in Epitopes A, B, C and D in HA1 mutated at a high frequency in global circulating strains last decade. The charged amino acid mutations in Epitopes A (T135K) has shown high mutability in strains near years, resulting in a decrease of NGT135-135. Both K158N and K160T not only involved mutations charged in epitope B, but also caused a gain of NYT158-160. Epitope B and its adjacent N-glycosylation site NYT158-160 mutated more frequently, which might be under greater immune pressure than the rest. The charged amino acid mutations in A/H3N2 Influenza play a significant role in virus evolution, which might cause an important public health issue. Variability related to both the epitopes (A and B) and N-glycosylation is beneficial for understanding the evolutionary mechanisms, disease pathogenesis and vaccine research.

  18. The special neuraminidase stalk-motif responsible for increased virulence and pathogenesis of H5N1 influenza A virus.

    Directory of Open Access Journals (Sweden)

    Hongbo Zhou

    Full Text Available The variation of highly pathogenic avian influenza H5N1 virus results in gradually increased virulence in poultry, and human cases continue to accumulate. The neuraminidase (NA stalk region of influenza virus varies considerably and may associate with its virulence. The NA stalk region of all N1 subtype influenza A viruses can be divided into six different stalk-motifs, H5N1/2004-like (NA-wt, WSN-like, H5N1/97-like, PR/8-like, H7N1/99-like and H5N1/96-like. The NA-wt is a special NA stalk-motif which was first observed in H5N1 influenza virus in 2000, with a 20-amino acid deletion in the 49(th to 68(th positions of the stalk region. Here we show that there is a gradual increase of the special NA stalk-motif in H5N1 isolates from 2000 to 2007, and notably, the special stalk-motif is observed in all 173 H5N1 human isolates from 2004 to 2007. The recombinant H5N1 virus with the special stalk-motif possesses the highest virulence and pathogenicity in chicken and mice, while the recombinant viruses with the other stalk-motifs display attenuated phenotype. This indicates that the special stalk-motif has contributed to the high virulence and pathogenicity of H5N1 isolates since 2000. The gradually increasing emergence of the special NA stalk-motif in H5N1 isolates, especially in human isolates, deserves attention by all.

  19. Influenza NA and PB1 Gene Segments Interact during the Formation of Viral Progeny: Localization of the Binding Region within the PB1 Gene

    Directory of Open Access Journals (Sweden)

    Brad Gilbertson

    2016-08-01

    Full Text Available The influenza A virus genome comprises eight negative-sense viral RNAs (vRNAs that form individual ribonucleoprotein (RNP complexes. In order to incorporate a complete set of each of these vRNAs, the virus uses a selective packaging mechanism that facilitates co-packaging of specific gene segments but whose molecular basis is still not fully understood. Recently, we used a competitive transfection model where plasmids encoding the A/Puerto Rico/8/34 (PR8 and A/Udorn/307/72 (Udorn PB1 gene segments were competed to show that the Udorn PB1 gene segment is preferentially co-packaged into progeny virions with the Udorn NA gene segment. Here we created chimeric PB1 genes combining both Udorn and PR8 PB1 sequences to further define the location within the Udorn PB1 gene that drives co-segregation of these genes and show that nucleotides 1776–2070 of the PB1 gene are crucial for preferential selection. In vitro assays examining specific interactions between Udorn NA vRNA and purified vRNAs transcribed from chimeric PB1 genes also supported the importance of this region in the PB1-NA interaction. Hence, this work identifies an association between viral genes that are co-selected during packaging. It also reveals a region potentially important in the RNP-RNP interactions within the supramolecular complex that is predicted to form prior to budding to allow one of each segment to be packaged in the viral progeny. Our study lays the foundation to understand the co-selection of specific genes, which may be critical to the emergence of new viruses with pandemic potential.

  20. First case of compound heterozygosity in Na,K-ATPase gene ATP1A2 in familial hemiplegic migraine.

    Science.gov (United States)

    Vanmolkot, Kaate R J; Stam, Anine H; Raman, Ashok; Koenderink, Jan B; de Vries, Boukje; van den Boogerd, Eelke H; van Vark, Judith; van den Heuvel, Jeroen J M W; Bajaj, Nin; Terwindt, Gisela M; Haan, Joost; Frants, Rune R; Ferrari, Michel D; van den Maagdenberg, Arn M J M

    2007-08-01

    Familial hemiplegic migraine (FHM) is a rare autosomal-dominant subtype of migraine with aura, associated with hemiparesis during the aura. Here we describe a unique FHM family in which two novel allelic missense mutations in the Na,K-ATPase gene ATP1A2 segregate in the proband with hemiplegic migraine. Both mutations show reduced penetrance in family members of the proband. Cellular survival assays revealed Na,K-ATPase dysfunction for both ATP1A2 mutants, indicating that both mutations are disease causative. This is the first case of compound heterozygosity for any of the known FHM genes.

  1. Dual Acting Neuraminidase Inhibitors Open New Opportunities to Disrupt the Lethal Synergism between Streptococcus pneumoniae and Influenza Virus

    Directory of Open Access Journals (Sweden)

    Elisabeth eWalther

    2016-03-01

    Full Text Available Secondary infections with Streptococcus pneumoniae cause severe pneumonia and enhance lethality during influenza epidemics and pandemics. Structural and functional similarities with viral neuraminidase (NA suggest that the highly prevalent pneumococcal NAs, NanA and NanB, might contribute to this lethal synergism by supporting viral replication and that dual acting NA inhibitors (NAIs will disrupt it. To verify this hypothesis, NanA and NanB were expressed in E. coli. After confirming their activity in enzyme assays, in vitro models with influenza virus A/Jena/8178/09 (Jena/8178 and the recombinant NanA or NanB (rNanA and rNanB were established in A549 and MDCK cells to mimic the role of these pneumococcal NAs during co-infection. Studies on the influence of both NAs on viral receptor expression, spread, and yield revealed a distinct effect of NanA and NanB on viral replication in these in vitro models. Both enzymes were able to support Jena/8178 replication at certain concentrations. This synergism was disrupted by the NAIs oseltamivir, DANA, katsumadain A, and artocarpin exerting an inhibitory effect on viral NA and NanA. Interestingly, katsumadain A and artocarpin inhibited rNanA and rNanB similarly. Zanamivir did not show activity. These results demonstrate a key role of pneumococcal NAs in the lethal synergism with influenza viruses and reveal opportunities for its effective disruption.

  2. QSAR analyses on avian influenza virus neuraminidase inhibitors using CoMFA, CoMSIA, and HQSAR

    Science.gov (United States)

    Zheng, Mingyue; Yu, Kunqian; Liu, Hong; Luo, Xiaomin; Chen, Kaixian; Zhu, Weiliang; Jiang, Hualiang

    2006-09-01

    The recent wide spreading of the H5N1 avian influenza virus (AIV) in Asia, Europe and Africa and its ability to cause fatal infections in human has raised serious concerns about a pending global flu pandemic. Neuraminidase (NA) inhibitors are currently the only option for treatment or prophylaxis in humans infected with this strain. However, drugs currently on the market often meet with rapidly emerging resistant mutants and only have limited application as inadequate supply of synthetic material. To dig out helpful information for designing potent inhibitors with novel structures against the NA, we used automated docking, CoMFA, CoMSIA, and HQSAR methods to investigate the quantitative structure-activity relationship for 126 NA inhibitors (NIs) with great structural diversities and wide range of bioactivities against influenza A virus. Based on the binding conformations discovered via molecular docking into the crystal structure of NA, CoMFA and CoMSIA models were successfully built with the cross-validated q 2 of 0.813 and 0.771, respectively. HQSAR was also carried out as a complementary study in that HQSAR technique does not require 3D information of these compounds and could provide a detailed molecular fragment contribution to the inhibitory activity. These models also show clearly how steric, electrostatic, hydrophobicity, and individual fragments affect the potency of NA inhibitors. In addition, CoMFA and CoMSIA field distributions are found to be in well agreement with the structural characteristics of the corresponding binding sites. Therefore, the final 3D-QSAR models and the information of the inhibitor-enzyme interaction should be useful in developing novel potent NA inhibitors.

  3. The cardiac sodium channel gene SCN5A and its gene product NaV1.5: Role in physiology and pathophysiology

    NARCIS (Netherlands)

    Veerman, Christiaan C.; Wilde, Arthur A. M.; Lodder, Elisabeth M.

    2015-01-01

    The gene SCN5A encodes the main cardiac sodium channel NaV1.5. This channel predominates the cardiac sodium current, INa, which underlies the fast upstroke of the cardiac action potential. As such, it plays a crucial role in cardiac electrophysiology. Over the last 60years a tremendous amount of

  4. Molecular characterization and functional analysis of a vacuolar Na(+)/H(+) antiporter gene (HcNHX1) from Halostachys caspica.

    Science.gov (United States)

    Guan, Bo; Hu, Youzhen; Zeng, Youling; Wang, Yan; Zhang, Fuchun

    2011-03-01

    According to sequences of several vacuolar Na(+)/H(+) antiporter genes from Xinjiang halophytic plants, a new vacuolar Na(+)/H(+) antiporter gene (HcNHX1) from the halophyte Halostachys caspica was obtained by RACE and RT-PCR using primers corresponding to conserved regions of the coding sequences. The obtained HcNHX1 cDNA was 1,983 bp and contained a 1,656 bp open reading frame encoding a deduced protein of 551 amino acid residues. The deduced amino acid sequence showed high identity with other NHX1 we have cloned previously from halophyte in Xinjiang desert area. The phylogenetic analysis showed that HcNHX1 formed a clade with NHX homologs of Chenopodiaceae. Expression profiles under salt treatment and ABA induction were investigated, and the results revealed that expression of HcNHX1 was induced by NaCl and ABA. To compare the degree of salt tolerance, we over-expressed HcNHX1 in Arabidopsis. Two transgenic lines grew more vigorously than the wild type (WT) under salt stress. The analysis of ion contents indicated that under salt stress, the transgenic plants compartmentalized more Na(+) in the leaves compared with wild-type plants. Together, these results suggest that the products of the novel gene HcNHX1 from halophyte Halostachys caspica is a functional tonoplast Na(+)/H(+) antiporter.

  5. Regeneration of membrane sialic acid after neuraminidase treatment of leukemic granulocytes.

    Science.gov (United States)

    Taub, R N; Hindenburg, A A; Baker, M A

    1985-01-01

    Granulocytes from patients with chronic myelogenous leukemia (CML) were studied for their ability to regenerate surface sialic acid following treatment with Vibrio cholera neuraminidase (VCN) in vitro. Immediately after neuraminidase treatment, CML and normal granulocytes showed similar incorporation of radioactivity after surface labelling with sodium periodate/potassium-H3-borohydride (PI/BH3(4)). CML granulocytes treated with neuraminidase then incubated for 18 h in nutrient medium showed strikingly increased PI/BH3(4) labelling, usually greater than initial pretreatment values, consistent with a rapid reappearance of sialic acid in the cell membrane. This pattern was not seen in normal granulocytes. The aberrant regeneration of sialic acid in CML granulocytes in vitro could be inhibited by addition of 3 X 10(-6) M retinoic acid, suggesting either a direct effect on membrane glycoconjugate synthesis or an association with granulocyte differentiation.

  6. Transforming growth factor-β: activation by neuraminidase and role in highly pathogenic H5N1 influenza pathogenesis.

    Directory of Open Access Journals (Sweden)

    Christina M Carlson

    2010-10-01

    Full Text Available Transforming growth factor-beta (TGF-β, a multifunctional cytokine regulating several immunologic processes, is expressed by virtually all cells as a biologically inactive molecule termed latent TGF-β (LTGF-β. We have previously shown that TGF-β activity increases during influenza virus infection in mice and suggested that the neuraminidase (NA protein mediates this activation. In the current study, we determined the mechanism of activation of LTGF-β by NA from the influenza virus A/Gray Teal/Australia/2/1979 by mobility shift and enzyme inhibition assays. We also investigated whether exogenous TGF-β administered via a replication-deficient adenovirus vector provides protection from H5N1 influenza pathogenesis and whether depletion of TGF-β during virus infection increases morbidity in mice. We found that both the influenza and bacterial NA activate LTGF-β by removing sialic acid motifs from LTGF-β, each NA being specific for the sialic acid linkages cleaved. Further, NA likely activates LTGF-β primarily via its enzymatic activity, but proteases might also play a role in this process. Several influenza A virus subtypes (H1N1, H1N2, H3N2, H5N9, H6N1, and H7N3 except the highly pathogenic H5N1 strains activated LTGF-β in vitro and in vivo. Addition of exogenous TGF-β to H5N1 influenza virus-infected mice delayed mortality and reduced viral titers whereas neutralization of TGF-β during H5N1 and pandemic 2009 H1N1 infection increased morbidity. Together, these data show that microbe-associated NAs can directly activate LTGF-β and that TGF-β plays a pivotal role protecting the host from influenza pathogenesis.

  7. A comparative gene analysis with rice identified orthologous group II HKT genes and their association with Na(+) concentration in bread wheat.

    Science.gov (United States)

    Ariyarathna, H A Chandima K; Oldach, Klaus H; Francki, Michael G

    2016-01-19

    Although the HKT transporter genes ascertain some of the key determinants of crop salt tolerance mechanisms, the diversity and functional role of group II HKT genes are not clearly understood in bread wheat. The advanced knowledge on rice HKT and whole genome sequence was, therefore, used in comparative gene analysis to identify orthologous wheat group II HKT genes and their role in trait variation under different saline environments. The four group II HKTs in rice identified two orthologous gene families from bread wheat, including the known TaHKT2;1 gene family and a new distinctly different gene family designated as TaHKT2;2. A single copy of TaHKT2;2 was found on each homeologous chromosome arm 7AL, 7BL and 7DL and each gene was expressed in leaf blade, sheath and root tissues under non-stressed and at 200 mM salt stressed conditions. The proteins encoded by genes of the TaHKT2;2 family revealed more than 93% amino acid sequence identity but ≤52% amino acid identity compared to the proteins encoded by TaHKT2;1 family. Specifically, variations in known critical domains predicted functional differences between the two protein families. Similar to orthologous rice genes on chromosome 6L, TaHKT2;1 and TaHKT2;2 genes were located approximately 3 kb apart on wheat chromosomes 7AL, 7BL and 7DL, forming a static syntenic block in the two species. The chromosomal region on 7AL containing TaHKT2;1 7AL-1 co-located with QTL for shoot Na(+) concentration and yield in some saline environments. The differences in copy number, genes sequences and encoded proteins between TaHKT2;2 homeologous genes and other group II HKT gene families within and across species likely reflect functional diversity for ion selectivity and transport in plants. Evidence indicated that neither TaHKT2;2 nor TaHKT2;1 were associated with primary root Na(+) uptake but TaHKT2;1 may be associated with trait variation for Na(+) exclusion and yield in some but not all saline environments.

  8. Proteome Response of Chicken Embryo Fibroblast Cells to Recombinant H5N1 Avian Influenza Viruses with Different Neuraminidase Stalk Lengths.

    Science.gov (United States)

    Li, Yongtao; Ming, Fan; Huang, Huimin; Guo, Kelei; Chen, Huanchun; Jin, Meilin; Zhou, Hongbo

    2017-01-12

    The variation on neuraminidase (NA) stalk region of highly pathogenic avian influenza H5N1 virus results in virulence change in animals. In our previous studies, the special NA stalk-motif of H5N1 viruses has been demonstrated to play a significant role in the high virulence and pathogenicity in chickens. However, the molecular mechanisms underlying the pathogenicity of viruses with different NA stalk remain poorly understood. This study presents a comprehensive characterization of the proteome response of chicken cells to recombinant H5N1 virus with stalk-short NA (rNA-wt) and the stalkless NA mutant virus (rSD20). 208 proteins with differential abundance profiles were identified differentially expressed (DE), and these proteins were mainly related to stress response, transcription regulation, transport, metabolic process, cellular component and cytoskeleton. Through Ingenuity Pathways Analysis (IPA), the significant biological functions of DE proteins represented included Post-Translational Modification, Protein Folding, DNA Replication, Recombination and Repair. It was interesting to find that most DE proteins were involved in the TGF-β mediated functional network. Moreover, the specific DE proteins may play important roles in the innate immune responses and H5N1 virus replication. Our data provide important information regarding the comparable host response to H5N1 influenza virus infection with different NA stalk lengths.

  9. Systematic review of influenza resistance to the neuraminidase inhibitors

    Directory of Open Access Journals (Sweden)

    Boivin Guy

    2011-05-01

    Full Text Available Abstract Background Antivirals play a critical role in the prevention and the management of influenza. One class of antivirals, neuraminidase inhibitors (NAIs, is effective against all human influenza viruses. Currently there are two NAI drugs which are licensed worldwide: oseltamivir (Tamiflu® and zanamivir (Relenza®; and two drugs which have received recent approval in Japan: peramivir and laninamivir. Until recently, the prevalence of antiviral resistance has been relatively low. However, almost all seasonal H1N1 strains that circulated in 2008-09 were resistant to oseltamivir whereas about 1% of tested 2009 pandemic H1N1 viruses were found to be resistant to oseltamivir. To date, no studies have demonstrated widespread resistance to zanamivir. It seems likely that the literature on antiviral resistance associated with oseltamivir as well as zanamivir is now sufficiently comprehensive to warrant a systematic review. The primary objectives were to systematically review the literature to determine the incidence of resistance to oseltamivir, zanamivir, and peramivir in different population groups as well as assess the clinical consequences of antiviral resistance. Methods We searched MEDLINE and EMBASE without language restrictions in September 2010 to identify studies reporting incidence of resistance to oseltamivir, zanamivir, and peramivir. We used forest plots and meta-analysis of incidence of antiviral resistance associated with the three NAIs. Subgroup analyses were done across a number of population groups. Meta-analysis was also performed to evaluate associations between antiviral resistance and clinical complications and symptoms. Results We identified 19 studies reporting incidence of antiviral resistance. Meta-analysis of 15 studies yielded a pooled incidence rate for oseltamivir resistance of 2.6% (95%CI 0.7% to 5.5%. The incidence rate for all zanamivir resistance studies was 0%. Only one study measured incidence of antiviral

  10. Chalcones as novel influenza A (H1N1) neuraminidase inhibitors from Glycyrrhiza inflata

    DEFF Research Database (Denmark)

    Dao, Trong Tuan; Nguyen, Phi Hung; Lee, Hong Sik

    2011-01-01

    -8) chalcones were isolated as active principles from the acetone extract of Glycyrrhiza inflata. Compounds 3 and 6 without prenyl group showed strong inhibitory effects on various neuraminidases from influenza viral strains, H1N1, H9N2, novel H1N1 (WT), and oseltamivir-resistant novel H1N1 (H274Y) expressed...

  11. Fitness costs for Influenza B viruses carrying neuraminidase inhibitor-resistant substitutions: underscoring the importance of E119A and H274Y.

    Science.gov (United States)

    Burnham, Andrew J; Baranovich, Tatiana; Marathe, Bindumadhav M; Armstrong, Jianling; Webster, Robert G; Govorkova, Elena A

    2014-05-01

    Influenza B viruses cause annual outbreaks of respiratory illness in humans and are increasingly recognized as a major cause of influenza-associated pediatric mortality. Neuraminidase (NA) inhibitors (NAIs) are the only available therapy for patients infected with influenza B viruses, and the potential emergence of NAI-resistant viruses is a public health concern. The NA substitutions located within the enzyme active site could not only reduce NAI susceptibility of influenza B virus but also affect virus fitness. In this study, we investigated the effect of single NA substitutions on the fitness of influenza B/Yamanashi/166/1998 viruses (Yamagata lineage). We generated recombinant viruses containing either wild-type (WT) NA or NA with a substitution in the catalytic (R371K) or framework (E119A, D198E, D198Y, I222T, H274Y, and N294S) residues. We assessed NAI susceptibility, NA biochemical properties, NA protein expression, and virus replication in vitro and in differentiated normal human bronchial epithelial (NHBE) cells. Our results showed that four NA substitutions (D198E, I222T, H274Y, and N294S) conferred reduced inhibition by oseltamivir and three (E119A, D198Y, and R371K) conferred highly reduced inhibition by oseltamivir, zanamivir, and peramivir. All NA substitutions, except for D198Y and R371K, were genetically stable after seven passages in MDCK cells. Cell surface NA protein expression was significantly increased by H274Y and N294S substitutions. Viruses with the E119A, I222T, H274Y, or N294S substitution were not attenuated in replication efficiency in vitro or in NHBE cells. Overall, viruses with the E119A or H274Y NA substitution possess fitness comparable to NAI-susceptible virus, and the acquisition of these substitutions by influenza B viruses should be closely monitored.

  12. Osmotic challenge and expression of aquaporin 3 and Na/K ATPase genes in bovine embryos produced in vitro.

    Science.gov (United States)

    Camargo, Luiz Sergio Almeida; Boite, Mariana Cortes; Wohlres-Viana, Sabine; Mota, Gustavo Bruno; Serapiao, Raquel Varela; Sa, Wanderlei Ferreira; Viana, Joao Henrique Moreira; Nogueira, Luiz Altamiro Garcia

    2011-12-01

    The aim of this study was to evaluate the effect of culture media and stage of development in the osmotic ability of in vitro-fertilized bovine embryos and the expression of aquaporin 3 (Aqp3) and Na/K ATPase isoform 1 (ATPAse1) genes in embryos (i) with different ability to undergo rehydration and (ii) following vitrification. In experiment 1, in vitro fertilized presumptive zygotes were co-cultured in SOFaac or modified CR2aa medium and embryos at blastocyst and expanded blastocyst stages at day 7 post-insemination were exposed to NaCl hypertonic medium (900 mOsm) for 5 min following 120 min of culture in isotonic medium in order to evaluate dehydration and rehydration, respectively. No difference (P>0.05) on blastocyst rate was found between CR2aa and SOFaac medium but embryos co-cultured in SOFaac medium underwent greater (PEmbryos at expanded blastocyst stage underwent greater dehydration but slower rehydration than embryos at blastocysts stage (P0.05) on relative amount of transcripts was found in either genes. In the experiment 3, expanded blastocysts produced in a co-culture system were vitrified, warmed and then cultured for 72 h for analysis of embryo survival and amount of Aqp3 and ATPase1 transcripts. Lower (Pembryo survival rate was found for vitrified-warmed embryos (57.9%) than for their fresh counterparts (84.6%). There was no difference on expression of ATPase1 gene but lower (Pembryos. In conclusion, embryo ability to undergo shrinkage and swelling is influenced by medium used in a co-culture system and by embryo stage. Rehydrating ability of embryos after exposure to NaCl hypertonic medium is not associated with variations on expression of Aqp3 and ATPase1 genes, but the vitrification can alter gene expression of in vitro-fertilized bovine embryos produced in a co-culture system. Copyright © 2011 Elsevier Inc. All rights reserved.

  13. Gene silencing reveals multiple functions of Na+/K+-ATPase in the salmon louse (Lepeophtheirus salmonis).

    Science.gov (United States)

    Komisarczuk, Anna Z; Kongshaug, Heidi; Nilsen, Frank

    2018-01-12

    Na+/K+-ATPase has a key function in a variety of physiological processes including membrane excitability, osmoregulation, regulation of cell volume, and transport of nutrients. While knowledge about Na+/K+-ATPase function in osmoregulation in crustaceans is extensive, the role of this enzyme in other physiological and developmental processes is scarce. Here, we report characterization, transcriptional distribution and likely functions of the newly identified L. salmonis Na+/K+-ATPase (LsalNa+/K+-ATPase) α subunit in various developmental stages. The complete mRNA sequence was identified, with 3003 bp open reading frame encoding a putative protein of 1001 amino acids. Putative protein sequence of LsalNa+/K+-ATPase revealed all typical features of Na+/K+-ATPase and demonstrated high sequence identity to other invertebrate and vertebrate species. Quantitative RT-PCR analysis revealed higher LsalNa+/K+-ATPase transcript level in free-living stages in comparison to parasitic stages. In situ hybridization analysis of copepodids and adult lice revealed LsalNa+/K+-ATPase transcript localization in a wide variety of tissues such as nervous system, intestine, reproductive system, and subcuticular and glandular tissue. RNAi mediated knock-down of LsalNa+/K+-ATPase caused locomotion impairment, and affected reproduction and feeding. Morphological analysis of dsRNA treated animals revealed muscle degeneration in larval stages, severe changes in the oocyte formation and maturation in females and abnormalities in tegmental glands. Thus, the study represents an important foundation for further functional investigation and identification of physiological pathways in which Na+/K+-ATPase is directly or indirectly involved. Copyright © 2018. Published by Elsevier Inc.

  14. (H5N1) strain neuraminidase expressed in yeast Pichia pastoris

    Indian Academy of Sciences (India)

    2014-03-20

    Mar 20, 2014 ... The colonies were subcultured on YPD broth and gene integration was confirmed as described at ww.invitrogen.com/content/sfs/manuals/pich_man.pdf. 2.4 Expression of rNA in Pichia pastoris. The positive transformants were inoculated into 2 mL of. Buffered Glycerol-complex Medium (BMGY) media and.

  15. Second Sialic Acid Binding Site in Newcastle Disease Virus Hemagglutinin-Neuraminidase: Implications for Fusion

    OpenAIRE

    Zaitsev, Viatcheslav; von Itzstein, Mark; Groves, Darrin; Kiefel, Milton; Takimoto, Toru; Portner, Allen; Taylor, Garry

    2004-01-01

    Paramyxoviruses are the leading cause of respiratory disease in children. Several paramyxoviruses possess a surface glycoprotein, the hemagglutinin-neuraminidase (HN), that is involved in attachment to sialic acid receptors, promotion of fusion, and removal of sialic acid from infected cells and progeny virions. Previously we showed that Newcastle disease virus (NDV) HN contained a pliable sialic acid recognition site that could take two states, a binding state and a catalytic state. Here we ...

  16. Microcapsules functionalized with neuraminidase can enter vascular endothelial cells in vitro.

    Science.gov (United States)

    Liu, Weizhi; Wang, Xiaocong; Bai, Ke; Lin, Miao; Sukhorukov, Gleb; Wang, Wen

    2014-12-06

    Microcapsules made of polyelectrolyte multilayers exhibit no or low toxicity, appropriate mechanical stability, variable controllable degradation and can incorporate remote release mechanisms triggered by various stimuli, making them well suited for targeted drug delivery to live cells. This study investigates interactions between microcapsules made of synthetic (i.e. polystyrenesulfonate sodium salt/polyallylamine hydrochloride) or natural (i.e. dextran sulfate/poly-L-arginine) polyelectrolyte and human umbilical vein endothelial cells with particular focus on the effect of the glycocalyx layer on the intake of microcapsules by endothelial cells. Neuraminidase cleaves N-acetyl neuraminic acid residues of glycoproteins and targets the sialic acid component of the glycocalyx on the cell membrane. Three-dimensional confocal images reveal that microcapsules, functionalized with neuraminidase, can be internalized by endothelial cells. Capsules without neuraminidase are blocked by the glycocalyx layer. Uptake of the microcapsules is most significant in the first 2 h. Following their internalization by endothelial cells, biodegradable DS/PArg capsules rupture by day 5; however, there is no obvious change in the shape and integrity of PSS/PAH capsules within the period of observation. Results from the study support our hypothesis that the glycocalyx functions as an endothelial barrier to cross-membrane movement of microcapsules. Neuraminidase-loaded microcapsules can enter endothelial cells by localized cleavage of glycocalyx components with minimum disruption of the glycocalyx layer and therefore have high potential to act as drug delivery vehicles to reach tissues beyond the endothelial barrier of blood vessels. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

  17. Financial conflicts of interest and conclusions about neuraminidase inhibitors for influenza: an analysis of systematic reviews.

    Science.gov (United States)

    Dunn, Adam G; Arachi, Diana; Hudgins, Joel; Tsafnat, Guy; Coiera, Enrico; Bourgeois, Florence T

    2014-10-07

    Industry funding and financial conflicts of interest may contribute to bias in the synthesis and interpretation of scientific evidence. To examine the association between financial conflicts of interest and characteristics of systematic reviews of neuraminidase inhibitors. Retrospective analysis. Reviews that examined the use of neuraminidase inhibitors in the prophylaxis or treatment of influenza, were published between January 2005 and May 2014, and used a systematic search protocol. Two investigators blinded to all information regarding the review authors independently assessed the presentation of evidence on the use of neuraminidase inhibitors as favorable or not favorable. Financial conflicts of interest were identified using the index reviews, other publications, and Web-based searches. Associations between financial conflicts of interest, favorability assessments, and presence of critical appraisals of evidence quality were analyzed. Twenty-six systematic reviews were identified, of which 13 examined prophylaxis and 24 examined treatment, accounting for 37 distinct assessments. Among assessments associated with a financial conflict of interest, 7 of 8 (88%) were classified as favorable, compared with 5 of 29 (17%) among those without a financial conflict of interest. Reviewers without financial conflicts of interest were more likely to include statements about the quality of the primary studies than those with financial conflicts of interest. The heterogeneity in populations and outcomes examined in the reviews precluded analysis of the contribution of selective inclusion of evidence on the discordance of the assessments made in the reviews. Many of the systematic reviews had overlapping authorship. Reviewers with financial conflicts of interest may be more likely to present evidence about neuraminidase inhibitors in a favorable manner and recommend the use of these drugs than reviewers without financial conflicts of interest. Australian National Health and

  18. Exploring the mechanism of zanamivir resistance in a neuraminidase mutant: a molecular dynamics study.

    Directory of Open Access Journals (Sweden)

    Nanyu Han

    Full Text Available It is critical to understand the molecular basis of the drug resistance of influenza viruses to efficiently treat this infectious disease. Recently, H1N1 strains of influenza A carrying a mutation of Q136K in neuraminidase were found. The new strain showed a strong Zanamivir neutralization effect. In this study, normal molecular dynamics simulations and metadynamics simulations were employed to explore the mechanism of Zanamivir resistance. The wild-type neuraminidase contained a 3(10 helix before the 150 loop, and there was interaction between the 150 and 430 loops. However, the helix and the interaction between the two loops were disturbed in the mutant protein due to interaction between K136 and nearby residues. Hydrogen-bond network analysis showed weakened interaction between the Zanamivir drug and E276/D151 on account of the electrostatic interaction between K136 and D151. Metadynamics simulations showed that the free energy landscape was different in the mutant than in the wild-type neuraminidase. Conformation with the global minimum of free energy for the mutant protein was different from the wild-type conformation. While the drug fit completely into the active site of the wild-type neuraminidase, it did not match the active site of the mutant variant. This study indicates that the altered hydrogen-bond network and the deformation of the 150 loop are the key factors in development of Zanamivir resistance. Furthermore, the Q136K mutation has a variable effect on conformation of different N1 variants, with conformation of the 1918 N1 variant being more profoundly affected than that of the other N1 variants studied in this paper. This observation warrants further experimental investigation.

  19. Aggregation of Streptococcus sanguis by a neuraminidase-sensitive component of serum and crevicular fluid.

    OpenAIRE

    Morris, E. J.; McBride, B. C.

    1983-01-01

    A number of strains of Streptococcus sanguis were found to aggregate in nonimmune serum and in crevicular fluid. All strains which aggregated in serum also aggregated in saliva, but some strains which aggregated in saliva did not aggregate in serum. Aggregation was destroyed by treatment of serum or crevicular fluid with neuraminidase and was inhibited by gangliosides. Treatment of serum with proteases reduced aggregating activity. Adsorption of serum to hydroxyapatite did not reduce the aggr...

  20. A importância do gene p53 na carcinogênese humana The importance of the p53 gene in human carcinogenesis

    Directory of Open Access Journals (Sweden)

    Agnes C. Fett-Conte

    2002-04-01

    Full Text Available Existem várias razões que justificam o título de "guardião do genoma" do gene P53. Seu envolvimento, direto ou indireto, tem sido observado na etiopatogenia de praticamente todas as neoplasias humanas, incluindo as leucemias e linfomas. Conhecer seus mecanismos de ação é fundamental para compreender os aspectos moleculares da carcinogênese. O presente trabalho apresenta uma revisão sobre as características deste gene e sua importância no diagnóstico, prognóstico e terapêutica, o que faz dele um alvo em potencial das estratégias de terapia gênica.There are several reasons which justify the name of 'guardian of the genome' given to the P53 gene. Its involvement either directly or indirectly has been observed in the pathology of practically all human neoplasias, including leukemia and lymphomas. Knowledge of its mechanisms of action is fundamental to understand molecular aspects of carcinogenesis. This work presents a revision of the characteristics of this gene and its importance in the diagnosis, prognosis and treatment and why this makes it a potential target for gene therapy strategies.

  1. Depression, Cytokine, and Cytokine by Treatment Interactions Modulate Gene Expression in Antipsychotic Naïve First Episode Psychosis.

    Science.gov (United States)

    Noto, Cristiano; Ota, Vanessa Kiyomi; Santoro, Marcos Leite; Gouvea, Eduardo Sauerbronn; Silva, Patricia Natalia; Spindola, Leticia Maria; Cordeiro, Quirino; Bressan, Rodrigo Affonseca; Gadelha, Ary; Brietzke, Elisa; Belangero, Sintia Iole; Maes, Michael

    2016-10-01

    In schizophrenia, genetic and environmental factors affect neurodevelopment and neuroprogressive trajectory. Altered expression of neuro-immune genes and increased levels of cytokines are observed, especially in patients with comorbid depression. However, it remains unclear whether circulating levels of cytokines and expression of these genes are associated, and how antipsychotic treatments impact this association. Relationships between messenger RNA (mRNA) expression of 11 schizophrenia-related genes and circulating levels of cytokines (interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α) were analyzed in 174 antipsychotic naïve first episode psychosis (FEP) and in 77 healthy controls. A subgroup of 72 patients was reassessed after treatment with risperidone. FEP patients were divided into those with and without depression. FEP patients with depression showed increased COMT expression and decreased NDEL1 expression. Increased IL-6 was associated with lowered AKT1 and DROSHA expression, while increased IL-10 was associated with increased NDEL1, DISC1, and MBP expression. IL-6 levels significantly increased the risperidone-induced expression of AKT1, DICER1, DROSHA, and COMT mRNA. The differential mRNA gene expression in FEP is largely associated with increased cytokine levels. While increased IL-6 may downregulate AKT-mediated cellular functions and dysregulate genes involved in microRNA (miRNA) machinery, increased IL-10 has neuroprotective properties. Increased IL-6 levels may prime the expression of genes (AKT1, DICER1, DROSHA, and COMT) in response to risperidone, suggesting that cytokine × treatment × gene interactions may improve cell function profiles. FEP patients with depression show a different gene expression profile reinforcing the theory that depression in FEP is a different phenotype.

  2. OPPORTUNITIES FOR APPLICATION OF THE NEURAMINIDASE INHIBITORS IN TREATMENT AND PREVENTION OF THE FLU

    Directory of Open Access Journals (Sweden)

    Yu.B. Belan

    2007-01-01

    Full Text Available Flu viruses cause yearly epidemics with the lesion of approximately 20% of population. during the increase of the flu sickness rate, it is necessary to take urgent anti epidemic and treatment steps aimed to reduce the spread of an infection as soon as possible and incorporating the application of the specific antiviral medications. The major aims for prescription of the specific antiviral medications in flu treatment are to reduce the duration and severity of the leading disease symptoms, risks of complications, as well as to prevent the lethal out comes. There were developed medications, effectively inhibiting the replication of B flu virus. At present, there are the 1st generation medications available — adamantane derivatives (amantadine and rimantadine and the 2nd generation medications — neuraminidase inhibitors (oseltamivir and zanamivir. The fast increase of the flu virus resistance towards adamantanes determine the necessity of a wider application of neuraminidase inhibitors, which are highly effective in respect flu viruses of A and B, as well as avian flu virus (h5n1.Key words: neuraminidase inhibitors, adamantanes, oseltamivir, zanamivir, h3n2, h5n1.

  3. 'a'-Position-mutated and G4-mutated hemagglutinin-neuraminidase proteins of Newcastle disease virus impair fusion and hemagglutinin-neuraminidase-fusion interaction by different mechanisms.

    Science.gov (United States)

    Chu, Fu-lu; Wen, Hong-ling; Zhang, Wen-qiang; Lin, Bin; Zhang, Yan; Sun, Cheng-xi; Ren, Gui-jie; Song, Yan-yan; Wang, Zhiyu

    2013-01-01

    To determine the effects of heptad repeat regions (HRs) and N-linked carbohydrate sites of the Newcastle disease virus hemagglutinin-neuraminidase (HN) protein on fusion of HN and fusion (F) proteins and HN-F interaction. We mutated six 'a' residues in the HRs and four asparagines in N-linked carbohydrate sites to alanine in the HN protein. A vaccinia-T7 RNA polymerase expression system was used to express HN cDNAs in BHK-21 cells to determine the HN functions. Deglycosylation was treated with PGNase F digestion. The formation of HN-F protein complexes was determined by the coimmunoprecipitation assay. Each HR-mutated protein interfered with fusion and the HN-F interaction. The G4-mutated protein not only impaired fusion and HN-F interaction but also decreased activities in cell fusion promotion, hemadsorption and neuraminidase. It is assumed that two different mechanisms for mutations in these two regions are responsible for the decreased fusion promotion activity and the reduced ability of interaction with F protein. Mutations in the HRs impair fusion and HN-F interaction by altering the transmission of a signal from the globular domain to the F-specific region in the stalk, but the G4 mutation modulates fusion and HN-F interaction by the misfolding of some important structures. Copyright © 2012 S. Karger AG, Basel.

  4. Droplet digital PCR to investigate quasi-species at codons 119 and 275 of the A(H1N1)pdm09 neuraminidase during zanamivir and oseltamivir therapies.

    Science.gov (United States)

    Abed, Yacine; Carbonneau, Julie; L'Huillier, Arnaud G; Kaiser, Laurent; Boivin, Guy

    2017-04-01

    The H275Y and E119D neuraminidase (NA) mutations constitute important molecular markers of resistance to NA inhibitors in A(H1N1) pdm09 viruses. We used reverse transcriptase-droplet digital PCR amplification (RT-ddPCR) to analyze quasi-species at codons 275 and 119 of the NA in A(H1N1) pdm09 viruses recovered from an immuncompromised patient who received oseltamivir and zanamivir therapies. RT-ddPCR assays detected and quantified H275Y and E119D mutations with an efficiency that was comparable to that of high throughput sequencing (HiSeq 2500 Illumina, San Diego, CA) technology. With its sensitivity and reproducibility, RT-ddPCR could be a reliable method for accurate detection and quantification of major NAI-resistance mutations in clinical settings. J. Med. Virol. 89:737-741, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  5. Replication of H9 influenza viruses in the human ex vivo respiratory tract, and the influence of neuraminidase on virus release.

    Science.gov (United States)

    Chan, Renee W Y; Chan, Louisa L Y; Mok, Chris K P; Lai, Jimmy; Tao, Kin P; Obadan, Adebimpe; Chan, Michael C W; Perez, Daniel R; Peiris, J S Malik; Nicholls, John M

    2017-07-24

    H9N2 viruses are the most widespread influenza viruses in poultry in Asia. We evaluated the infection and tropism of human and avian H9 influenza virus in the human respiratory tract using ex vivo respiratory organ culture. H9 viruses infected the upper and lower respiratory tract and the majority of H9 viruses had a decreased ability to release virus from the bronchus rather than the lung. This may be attributed to a weak neuraminidase (NA) cleavage of carbon-6-linked sialic acid (Sia) rather than carbon-3-linked Sia. The modified cleavage of N-acetlylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) by NA in H9 virus replication was observed by reverse genetics, and recombinant H9N2 viruses with amino acids (38KQ) deleted in the NA stalk, and changing the amino acid at position 431 from Proline-to-Lysine. Using recombinant H9 viruses previously evaluated in the ferret, we found that viruses which replicated well in the ferret did not replicate to the same extent in the human ex vivo cultures. The existing risk assessment models for H9N2 viruses in ferrets may not always have a strong correlation with the replication in the human upper respiratory tract. The inclusion of the human ex vivo cultures would further strengthen the future risk-assessment strategies.

  6. The Goldfish SG2NA Gene Encodes Two α-Type Regulatory Subunits for PP-2A and Displays Distinct Developmental Expression Pattern

    Directory of Open Access Journals (Sweden)

    Hai-Li Ma

    2009-07-01

    Full Text Available SG2NA is a member of the striatin protein family. In human and mouse, the SG2NA gene encodes two major protein isoforms: SG2NAα and SG2NAβ. The functions of these proteins, except for acting as the regulatory subunits for PP-2A, remain largely unknown. To explore the possible functions of SG2NA in lower vertebrates, we have isolated two SG2NA cDNAs from goldfish, Carassius auratus. Our results reveal that the first cDNA contains an ORF of 2118 bp encoding a deduced protein with 705 amino acids, and the second one 2148 bp coding for a deduced protein of 715 amino acids. Comparative analysis reveals that both isoforms belong to the α-type, and are named SG2NAα and SG2NAα+. RT-PCR and western blot analysis reveal that the SG2NA gene is differentially expressed in 9 tissues examined. During goldfish development, while the SG2NA mRNAs remain relatively constant in the first 3 stages and then become decreased and fluctuated from gastrula to larval hatching, the SG2NA proteins are fluctuated, displaying a peak every 3 to 4 stages. Each later peak is higher than the earlier one and the protein expression level becomes maximal at hatching stage. Together, our results reveal that SG2NA may play an important role during goldfish development and also in homeostasis of most adult tissues.

  7. The Goldfish SG2NA Gene Encodes Two α-Type Regulatory Subunits for PP-2A and Displays Distinct Developmental Expression Pattern

    Directory of Open Access Journals (Sweden)

    Hai-Li Ma

    2009-01-01

    Full Text Available SG2NA is a member of the striatin protein family. In human and mouse, the SG2NA gene encodes two major protein isoforms: SG2NAα and SG2NAβ. The functions of these proteins, except for acting as the regulatory subunits for PP-2A, remain largely unknown. To explore the possible functions of SG2NA in lower vertebrates, we have isolated two SG2NA cDNAs from goldfish, Carassius auratus. Our results reveal that the first cDNA contains an ORF of 2118 bp encoding a deduced protein with 705 amino acids, and the second one 2148 bp coding for a deduced protein of 715 amino acids. Comparative analysis reveals that both isoforms belong to the α-type, and are named SG2NAα and SG2NAα+. RT-PCR and western blot analysis reveal that the SG2NA gene is differentially expressed in 9 tissues examined. During goldfish development, while the SG2NA mRNAs remain relatively constant in the first 3 stages and then become decreased and fluctuated from gastrula to larval hatching, the SG2NA proteins are fluctuated, displaying a peak every 3 to 4 stages. Each later peak is higher than the earlier one and the protein expression level becomes maximal at hatching stage. Together, our results reveal that SG2NA may play an important role during goldfish development and also in homeostasis of most adult tissues.

  8. Zanamivir immobilized magnetic beads for voltammetric measurement of neuraminidase at gold-modified boron doped diamond electrode

    Energy Technology Data Exchange (ETDEWEB)

    Wahyuni, Wulan Tri, E-mail: wulantriws@gmail.com [Department of Chemistry, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Kampus IPB Darmaga, Bogor 16680 (Indonesia); Department of Chemistry, FMIPA, Universitas Indonesia, Kampus UI Depok (Indonesia); Ivandini, Tribidasari A.; Saepudin, Endang [Department of Chemistry, FMIPA, Universitas Indonesia, Kampus UI Depok (Indonesia); Einaga, Yasuaki [Department of Chemistry, Faculty of Science and Technology, Keio University, Hiyoshi 3-14-1, Yokohama 223-8522 (Japan); CREST, JST, 3-14-1 Hiyoshi, Yokohama 223-8522 (Japan)

    2016-04-19

    Biomolecule modified magnetic beads has been widely used in separation and sensing process. This study used streptavidin modified magnetic beads to immobilize biotin modified zanamivir. Biotin-streptavidin affinity facilitates immobilization of zanamivir on magnetic beads. Then interaction of zanamivir and neuraminidase was adopted as basic for enzyme detection. Detection of neuraminidase was performed at gold modified BDD using cyclic voltammetry technique. The measurement was carried out based on alteration of electrochemical signals of working electrode as neuraminidase response. The result showed that zanamivir was successfully immobilized on magnetic beads. The optimum amount of magnetic beads for zanamivir immobilization was 120 ug. Linear responses of neuraminidase were detected in concentration range of 0-15 mU. Detection limit (LOD) of measurement was 2.32 mU (R2 = 0.959) with precision as % RSD of 1.41%. Measurement of neuraminidase on magnetic beads could be also performed in the presence of mucin matrix. The linearity range was 0-8 mU with LOD of 0.64 mU (R2 = 0.950) and % RSD of 7.25%.

  9. Intestinal neuraminidase activity of suckling rats and other mammals. Relationship to the sialic acid content of milk.

    Science.gov (United States)

    Dickson, J J; Messer, M

    1978-02-15

    1. The neuraminidase activity of homogenates of the mucosa of the middle and distal thirds of the small intestine of rats increased about 5-fold between birth and 4 to 8 days of age, and then gradually declined to the much lower adult activity by 24 days. No comparable changes occurred in the proximal third. 2. In 8-day-old rats, the neuraminidase activity of the middle and distal thirds of the small intestine was about 10 times greater than that of the proximal third, 20 times greater than that of the colon and at least 100 times greater than that of the liver, brain, gastric mucosa or pancreas. 3. In all other species investigated (mice, rabbits, cats and guinea pigs), the neuraminidase activity of the middle and distal thirds of the small intestine was greater in suckling animals than in adults. 4. The sialic acid content of rat milk increased about 2-fold between birth and 8 days post partum and then declined. 5. There was a highly significant positive correlation between the intestinal neuraminidase activity of suckling animals of various species and ages and the sialic acid content of milk obtained from the corresponding species and stage of lactation. 6. It is suggested that the intestinal neuraminidase of suckling mammals functions primarily to remove sialic acid from various components of milk, thus providing sialic acid for the synthesis of sialoglycoproteins and gangliosides by the young.

  10. In silico molecular modeling of neuraminidase enzyme H1N1 avian influenza virus and docking with zanamivir ligands

    Directory of Open Access Journals (Sweden)

    Muthiyan Ramachandran

    2012-12-01

    Full Text Available Objective: Neuraminidase is an enzyme aspartic protease that is essential for the life cycle of H1N1. Methods: Constructed a model of Neuraminidase enzyme the 3D structure as template using with Modeller software. The Neuraminidase enzyme model was predicted and validated by Procheck, What check, Errat, Verify-3D and AutoDock web server for reliability. Results: The Modeller homology-modeling algorithm was demonstrated excellent accuracy in blind predictions. The Neuraminidase enzyme model built with little, 35% identity could be accurate enough to be successfully used in receptor based rational drug design. The closest homologue with the highest sequence identity 100% was selected. Zanamivir drug and analogues were retrieved from PubChem database, as well as subjected to docking interaction with Neuraminidase enzyme used AutoDock programme. Based on the root mean square deviation and lowest binding energy values the best docking orientation was selected. The better lowest binding energy value -6.91 was selected of CID_25209232. Conclusions: This study will be used in broad screening of inhibitors of the protein. However, further implemented experimental and clinical verification is needed to establishment these analogues as drug.

  11. Avian adeno-associated virus-based expression of Newcastle disease virus hemagglutinin-neuraminidase protein for poultry vaccination.

    Science.gov (United States)

    Perozo, F; Villegas, P; Estevez, C; Alvarado, I R; Purvis, L B; Saume, E

    2008-06-01

    The avian adeno-associated virus (AAAV) is a replication-defective nonpathogenic virus member of the family Parvoviridae that has been proved to be useful as a viral vector for gene delivery. The use of AAAV for transgenic expression of Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) protein and its ability to induce immunity in chickens were assessed. Proposed advantages of this system include no interference with maternal antibodies, diminished immune response against the vector, and the ability to accommodate large fragments of genetic information. In this work the generation of recombinant AAAV virions expressing the HN protein (rAAAV-HN) was demonstrated by electron microscopy, immunocytochemistry, and western blot analysis. Serological evidence of HN protein expression after in ovo or intramuscular inoculation of the recombinant virus in specific-pathogen-free chickens was obtained. Serum from rAAAV-HN-vaccinated birds showed a systemic immune response evidenced by NDV-specific enzyme-linked immunosorbent assay and hemagglutination inhibition testing. Positive virus neutralization in embryonated chicken eggs and indirect immunofluorescence detection of NDV infected cells by serum from rAAAV-HN vaccinated birds is also reported. A vaccine-challenge experiment in commercial broiler chickens using a Venezuelan virulent viscerotropic strain of NDV was performed. All unvaccinated controls died within 5 days postchallenge. Protection up to 80% was observed in birds vaccinated in ovo and revaccinated at 7 days of age with the rAAAV-HN. The results demonstrate the feasibility of developing and using an AAAV-based gene delivery system for poultry vaccination.

  12. NaCl-induced expression of AtVHA-c5 gene in the roots plays a role in response of Arabidopsis to salt stress.

    Science.gov (United States)

    Zhou, Aimin; Liu, Enhui; Ma, Hongping; Feng, Shuang; Gong, Shufang; Wang, Jingang

    2018-01-06

    Suppression of AtVHA-c5 expression results in changes in H+ and Na+ fluxes of roots, and increase sensitivity to salt in Arabidopsis. Vacuolar-type H+-ATPase (V-ATPase), a multisubunit endomembrane proton pump, is essential in plant growth and response to environmental stresses. In the present study, the function of Arabidopsis V-ATPase subunit c5 (AtVHA-c5) gene in response to salt stress was investigated. Subcellular localization showed that AtVHA-c5 was mainly localized to endosomes and the vacuolar membrane in Arabidopsis. The analysis of quantitative real-time PCR showed that expression of AtVHA-c5 gene was induced by NaCl stress. Histochemical analysis revealed that AtVHA-c5 was expressed in the root epidermis of untreated Arabidopsis and in the whole root elongation zone after NaCl treatment. Phenotypic analysis showed that the atvha-c5 mutant is sensitive to high NaCl as compared to the wild type. The non-invasive micro-test technology measurement demonstrated that the net H+ and Na+ efflux in the root elongation zone of the atvha-c5 mutant was weaker than that of the wild type under NaCl treatment, suggesting that H+ and Na+ fluxes in atvha-c5 roots are impaired under NaCl stress. Moreover, compared to the wild type, the expression of AtSOS1 (salt overly sensitive 1) and AtAHA1 (plasma membrane H+-ATPase 1) were down-regulated in atvha-c5 roots under NaCl stress. Overall, our results indicate that AtVHA-c5 plays a role in Arabidopsis root response to NaCl stress by influencing H+ and Na+ fluxes.

  13. Newcastle Disease Virus Establishes Persistent Infection in Tumor Cells In Vitro: Contribution of the Cleavage Site of Fusion Protein and Second Sialic Acid Binding Site of Hemagglutinin-Neuraminidase.

    Science.gov (United States)

    Rangaswamy, Udaya S; Wang, Weijia; Cheng, Xing; McTamney, Patrick; Carroll, Danielle; Jin, Hong

    2017-08-15

    Newcastle disease virus (NDV) is an oncolytic virus being developed for the treatment of cancer. Following infection of a human ovarian cancer cell line (OVCAR3) with a recombinant low-pathogenic NDV, persistent infection was established in a subset of tumor cells. Persistently infected (PI) cells exhibited resistance to superinfection with NDV and established an antiviral state, as demonstrated by upregulation of interferon and interferon-induced genes such as myxoma resistance gene 1 (Mx1) and retinoic acid-inducing gene-I (RIG-I). Viruses released from PI cells induced higher cell-to-cell fusion than the parental virus following infection in two tumor cell lines tested, HT1080 and HeLa, and remained attenuated in chickens. Two mutations, one in the fusion (F) protein cleavage site, F117S (F117S), and another in hemagglutinin-neuraminidase (HN), G169R (HN169R), located in the second sialic acid binding region, were responsible for the hyperfusogenic phenotype. F117S improves F protein cleavage efficiency, facilitating cell-to-cell fusion, while HN169R possesses a multifaceted role in contributing to higher fusion, reduced receptor binding, and lower neuraminidase activity, which together result in increased fusion and reduced viral replication. Thus, establishment of persistent infection in vitro involves viral genetic changes that facilitate efficient viral spread from cell to cell as a potential mechanism to escape host antiviral responses. The results of our study also demonstrate a critical role in the viral life cycle for the second receptor binding region of the HN protein, which is conserved in several paramyxoviruses.IMPORTANCE Oncolytic Newcastle disease virus (NDV) could establish persistent infection in a tumor cell line, resulting in a steady antiviral state reflected by constitutively expressed interferon. Viruses isolated from persistently infected cells are highly fusogenic, and this phenotype has been mapped to two mutations, one each in the

  14. Comparison of immunological characteristics of peripheral, splenic and tonsilar naïve B cells by differential gene expression meta-analyses.

    Science.gov (United States)

    Chokeshai-u-saha, Kaj; Lepoivre, Cyrille; Grieco, Luca; Nguyen, Catherine; Ruxrungtham, Kiat

    2012-12-01

    Naïve B cells isolated from peripheral blood, spleen and tonsil are commonly used in human B cell studies. However, little has been written about their possible variations in immunological properties. This study compared differential gene expression in human naive B subsets by meta-analysis using expression data available in Gene Expression Onimbus (GEO). Gene expression files of the Affymetrix Human Genome U133A Array (Affymetrix) were downloaded to collect 21 total array data samples of peripheral naïve B cells (n=10), splenic naïve B cells (n=2), tonsilar naïve B cells (n=3), peripheral memory B cells (n=4) and splenic memory B cells (n=2). Prior to differential gene expression analyses, data were normalized in order to reduce non-biological variation among the datasets. Comparisons of peripheral naive B cells with their splenic and tonsilar counterparts showed remarkable differences in terms of gene expression (29 and 202 genes, respectively). However, only minor differences were detected between splenic and tonsilar naive B cells (10 genes), consistent with the clustering results classifying both of them as lymphoid naive B cells. Differential gene expression results also implied higher stimulating states of lymphoid naive B cells when compared with peripheral blood naive B cells. These included enhanced expressions of CD27, CR2, EGR1, GADD45B, ICAM1, ICOSLG, IGHA, IL6, MMP9, SAMSN1, SMAD7, TNFAIP3, but reduced HLA-DOB expression. Our findings suggest that results generated from peripheral naive B cells may not always be applicable to the biological activities of other lymphoid naïve B cells. Nonetheless, further biological study is warranted.

  15. Neuraminidase production by a Streptococcus sanguis strain associated with subacute bacterial endocarditis.

    Science.gov (United States)

    Straus, D C; Portnoy-Duran, C

    1983-01-01

    The properties of an extracellular neuraminidase produced by a Streptococcus sanguis strain (isolated from a confirmed case of subacute bacterial endocarditis) during growth in a defined medium was examined in this investigation. This enzyme, isolated from concentrated culture supernatants of S. sanguis biotype II, was active against human alpha-1 acid glycoprotein, N-acetylneuramin lactose, bovine submaxillary mucin, and fetuin. Neuraminidase production paralleled bacterial growth in defined medium and was maximal in the early stationary phase of growth but decreased dramatically, probably owing to protease production, during the late stationary phase. The enzyme was purified to near homogeneity by a combination of salt fractionation, ion-exchanged chromatography on DEAE-Sephacel, and gel filtration on Sephadex G-200. These procedures yielded an enzyme preparation that possessed a specific activity of 174.4 mumol of sialic acid released per min per mg of protein against human alpha-1 acid glycoprotein. The Km value for this enzyme with human alpha-1 acid glycoprotein as substrate was 2.5 X 10(-3) M, and the enzyme possessed a pH optimum of 6.5. The S. sanguis neuraminidase had a molecular weight of approximately 85,000 as estimated by gel filtration and approximately 90,000 when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was stable at temperatures of 4 and 37 degrees C for 3 h, but approximately 50% of the enzymatic activity was lost within 30 min at 50 degrees C, with 100% of the enzymatic activity being destroyed within 10 min at temperatures of greater than or equal to 65 degrees C. Images PMID:6874067

  16. Na+ Channel Scn1b Gene Regulates Dorsal Root Ganglion Nociceptor Excitability in Vivo*

    Science.gov (United States)

    Lopez-Santiago, Luis F.; Brackenbury, William J.; Chen, Chunling; Isom, Lori L.

    2011-01-01

    Nociceptive dorsal root ganglion (DRG) neurons express tetrodotoxin-sensitive (TTX-S) and -resistant (TTX-R) Na+ current (INa) mediated by voltage-gated Na+ channels (VGSCs). In nociceptive DRG neurons, VGSC β2 subunits, encoded by Scn2b, selectively regulate TTX-S α subunit mRNA and protein expression, ultimately resulting in changes in pain sensitivity. We hypothesized that VGSCs in nociceptive DRG neurons may also be regulated by β1 subunits, encoded by Scn1b. Scn1b null mice are models of Dravet Syndrome, a severe pediatric encephalopathy. Many physiological effects of Scn1b deletion on CNS neurons have been described. In contrast, little is known about the role of Scn1b in peripheral neurons in vivo. Here we demonstrate that Scn1b null DRG neurons exhibit a depolarizing shift in the voltage dependence of TTX-S INa inactivation, reduced persistent TTX-R INa, a prolonged rate of recovery of TTX-R INa from inactivation, and reduced cell surface expression of Nav1.9 compared with their WT littermates. Investigation of action potential firing shows that Scn1b null DRG neurons are hyperexcitable compared with WT. Consistent with this, transient outward K+ current (Ito) is significantly reduced in null DRG neurons. We conclude that Scn1b regulates the electrical excitability of nociceptive DRG neurons in vivo by modulating both INa and IK. PMID:21555511

  17. Isolation and preliminary function analysis of a Na /H antiporter gene ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-05

    Oct 5, 2009 ... Southern blot analysis detected multiple copies of MzNHX1 in the M. zumi genome. Northern blot analysis showed negligible expression of the gene in roots, but expression was detected in stems and ... RhNHX1 from rose, cNHX1 from citrus, and AtNHX1 from Arabidopsis thaliana. However, MzNHX1.

  18. Single electrode genosensor for simultaneous determination of sequences encoding hemagglutinin and neuraminidase of avian influenza virus type H5N1.

    Science.gov (United States)

    Grabowska, Iwona; Malecka, Kamila; Stachyra, Anna; Góra-Sochacka, Anna; Sirko, Agnieszka; Zagórski-Ostoja, Włodzimierz; Radecka, Hanna; Radecki, Jerzy

    2013-11-05

    The duo-genosensor consisting of two different oligonucleotide probes immobilized covalently on the surface of one gold electrode via Au-S bond formation was used for simultaneous determination of two different oligonucleotide targets. One of the probes, decorated on its 5'-end with ferrocene (SH-ssDNA-Fc), is complementary to the cDNA representing a sequence encoding part of H5 hemagglutinin from H5N1 virus. The second probe, decorated on its 5'-end with methylene blue (SH-ssDNA-MB), is complementary to cDNA representing the fragment of N1 neuraminidase from the same virus. The presence of both probes on the surface of gold electrodes was confirmed with Osteryoung square-wave voltammetry (OSWV). The changes in redox activity of both redox active complexes before and after the hybridization process were used as analytical signal. The peak at +400 ± 2 mV was observed in the presence of 40 nM ssDNA used as a target for SH-ssDNA-Fc probe. This peak increased with the increase of concentration of target ssDNA. It indicates the "signal on" mode of analytical signal generation. The peak at -250 ± 4 mV, characteristic for SH-ssDNA-MB probe, was decreasing with the increase of the concentration of the complementary ssDNA target starting from 8 to 100 nM. This indicates the generation of electrochemical signal according to the "signal off" mode. The proposed duo-genosensor is capable of simultaneous, specific, and good sensitivity probing for the sequences derived from genes encoding two main markers of the influenza virus, hemagglutinin and neuraminidase.

  19. Neuraminidase inhibitory polyketides from the marine-derived fungus Phoma herbarum.

    Science.gov (United States)

    Zhang, Gao Fei; Han, Wen Bo; Cui, Jiang Tao; Ng, Seik Weng; Guo, Zhi Kai; Tan, Ren Xiang; Ge, Hui Ming

    2012-01-01

    Two new polyketides, arthropsadiol C (1) and massarilactone H (2), together with six known derivatives (3-8) were isolated from the culture broth of the marine-derived fungus Phoma herbarum. Their structures were elucidated on the basis of spectroscopic methods, including 2D NMR techniques. Compounds 2, 4, 5, and 8 showed moderate neuraminidase inhibitory activity with IC(50) values ranging from 4.15 to 9.16 µM. © Georg Thieme Verlag KG Stuttgart · New York.

  20. The equilibrium constant for the interaction between a monoclonal Fab fragment and an influenza virus neuraminidase.

    Science.gov (United States)

    Jackson, D C; Howlett, G J; Nestorowicz, A; Webster, R G

    1983-03-01

    The affinity or equilibrium constant between an Fab fragment derived from monoclonal IgG directed against influenza virus neuraminidase was measured as 4.1 X 10(7) M-1. The method, which makes use of an air-driven ultracentrifuge, is simple and uses extremely small amounts (10(-11) mol) of material. Furthermore, interpretation of the data is based on sound theoretical considerations. The technique also allows m.w. of the interacting species to be measured and the stoichiometry of the reaction to be determined.

  1. Polymorphisms of HIV RT gene among the ART naïve native drug exposed rural PLHA

    Directory of Open Access Journals (Sweden)

    K Mohana Krishnan

    2012-01-01

    Full Text Available Background: The number of people living with human immunodeficiency virus (HIV is increasing day by day in India. The disease has now spread from urban areas to rural areas. The proof reading of the reverse transcriptase enzyme is poor, which may lead to genetic diversity within the HIV strains, which in turn leads to problems like failure or resistance in antiretroviral treatment. This study is designed to find out the polymorphisms of the reverse transcriptase gene of HIV, after the native drug pressure among antiretroviral therapy (ART naïve rural people living with HIV/AIDS (RPLHA. Materials and Methods : A total of 207 HIV-Reactive patients were allowed to take native drugs from the local area and were advised to attend the center for HIV after six months for a follow-up. At the time of the follow-up visit, a second blood sample was taken from 20 reactive native-drug exposed ART-naïve patients. The plasma was separated and transported at 20°C to the YRG Care Center for genotyping. Results: Among the 20 HIV-reactive samples processed for gene sequencing analysis to detect the genotypic variations, only one sample (5% showed high-level mutational resistance variations and the predominant polymorphisms detected were V35T (100%, K122E (94.44%, and V60I (88.88%. Conclusions: The presence of drug-resistance mutations, although minimal, was important, as the drug-resistant strains could spread among the RPLHA and to their sexual partners. There was a definite need to generate a drug resistance database and the polymorphic pattern of Indian strains concern to the future clinical management of the disease, and a vaccine design to contain the disease.

  2. Mutations to A/Puerto Rico/8/34 PB1 gene improves seasonal reassortant influenza A virus growth kinetics.

    Science.gov (United States)

    Plant, Ewan P; Liu, Teresa M; Xie, Hang; Ye, Zhiping

    2012-12-17

    It is desirable for influenza vaccine virus strains to have phenotypes that include good growth and hemagglutinin (HA) protein yield. The quality of these characteristics varies among the vaccine viruses and is usually due to multigenic effects. Many influenza A virus vaccine viruses are made as reassortants of the high yield virus A/Puerto Rico/8/34 (PR/8) and a circulating seasonal virus. Co-infection of eggs with the two viruses, and selection of reassortants with the HA and neuraminidase (NA) segments from the seasonal virus, can result in viruses that contain a mixture of internal genes derived from both the high yield virus and the circulating virus. Segment 2 (PB1), which encodes the RNA-dependent RNA polymerase, frequently cosegregates with the seasonal HA and NA segments. We asked whether mutations based on the seasonal PB1 genes could improve vaccine virus strains. Here we report that mutations to the PR/8 PB1 gene, based on differences observed between seasonal and PR/8 PB1 genes, accelerate egg and cell culture based replication for a reassortant virus containing HA and NA segments from the low yield A/Wyoming/03/2003 (H3N2) vaccine virus. Published by Elsevier Ltd.

  3. An Na+/H+ antiporter gene from wheat plays an important role in ...

    Indian Academy of Sciences (India)

    PRAKASH KUMAR

    Two yeast (Saccharomyces cerevisiae) strains, W303 (ura3-. 1, can-1-100, ... were grown in SD-ura medium until they reached an. OD600 of .... induce TaNHX2 overexpression in a galactose-induced medium. The apparent expression of the TaNHX2 gene in the min-dextrose YPD culture was likely to be due to baseline ...

  4. Effects of cortisol and salinity acclimation on Na+/K+/2Cl–- cotransporter gene expression and Na+, K+-ATPase activity in the gill of Persian sturgeon, Acipenser persicus, fry

    Directory of Open Access Journals (Sweden)

    Saber Khodabandeh

    2009-10-01

    Full Text Available Na+, K+-ATPase activity and Na+/K+/2Cl–- cotransporter (NKCC gene expression in the gills of Persian sturgeon, Acipenser persicus, fry (2-3 g, 3.30-8.12 cm total body length in freshwater (control group, diluted Caspian Sea water (5 ppt and after treatment with cortisol in freshwater were studied. Na+, K+-ATPase activity was lower in the 5 ppt-acclimated fish (1.07±0.05 _mol Pi/mg protein/h than in the control fish (1.19±0.05 μmol Pi/mg protein/h but this difference was not significant. nKCC gene expression in the 5 ppt-acclimated fish (1.6±0.07 was significantly higher than in the control fish (0.8±0.00. In the cortisol treated fish, Na+, K+-ATPase activity (1.91±0.05 μmol Pi/mg protein/h and NKCC gene expression (3.2±0.1 were significantly higher than in the control group. our results show that Persian sturgeon fry (2-3 g can tolerate 5 ppt salinity by changing their enzymatic content and activity, and that exogenous cortisol application can increase the osmoregulatory capacity of fry before release into brackish water and can reduce their mortality.

  5. Susceptibility of influenza viruses circulating in Western Saudi Arabia to neuraminidase inhibitors

    Directory of Open Access Journals (Sweden)

    Ahmed M. Tolah

    2016-04-01

    Full Text Available Objectives: To investigate the sensitivity of circulating influenza viruses in Western Saudi Arabia to neuraminidase inhibitors (NAIs; mainly, zanamivir and oseltamivir. Methods: Respiratory samples were collected from patients presenting with respiratory symptoms to King Abdulaziz University Hospital, Jeddah, Kingdom of Saudi Arabia (KSA between September 2013 and October 2014. All samples were tested prospectively by real-time reverse-transcription polymerase chain reaction for influenza A and B viruses. Positive samples were then inoculated on Madin-Darby Canine Kidney (MDCK cells and isolated viruses were examined for their sensitivity to NAIs using fluorescent neuraminidase inhibition assay. Results: Out of 406 tested samples, 25 samples (6.2% were positive for influenza A/pdmH1N1 virus, one sample (0.25% was positive for influenza A/H3N2 virus, and 7 samples (1.7% were positive for influenza B Yamagata-like virus. Screening of isolated influenza A and B viruses (9 out of 33 for their sensitivity to NAIs showed no significant resistance to available NAIs. Conclusion: Our results show that circulating influenza viruses in Jeddah are still sensitive to NAIs.

  6. Assignment of the human Na[sup +]/glucose cotransporter gene SGLT1 to chromosome 22q13. 1

    Energy Technology Data Exchange (ETDEWEB)

    Turk, E.; Klisak, I.; Bacallao, R.; Sparkes, R.S.; Wright, E.M. (UCLA School Medicine, Los Angeles, CA (United States))

    1993-09-01

    The Na[sup +]/glucose cotransporter gene SGLT1 encodes the primary carrier protein responsible for the uptake of the dietary sugars glucose and galactose from the intestinal lumen. SGLT1 transport activity is currently exploited in oral rehydration therapy. The 75-kDa glycoprotein is localized in the brush border of the intestinal epithelium and is predicted to comprise 12 membrane spans. In two patients with the autosomal recessive disease glucose/galactose malabsorption, the underlying cause was found to be a missense mutation in SGLT1, and the Asp28 [yields] Asn change was demonstrated in vitro to eliminate SGLT1 transport activity. The SGLT1 gene was previously shown to reside on the distal q arm of chromosome 22(11.2 [yields] qter). The authors have used a cosmid probe for fluorescence in situ hybridization, which refines the localization to 22q13.1, and provide an example of the utility of the SGLT1 probe as a diagnostic for genetic diseases associated with trans-locations of chromosome 22. 18 refs., 2 figs.

  7. Analysis of Anti-Influenza Virus Neuraminidase Antibodies in Children, Adults, and the Elderly by ELISA and Enzyme Inhibition: Evidence for Original Antigenic Sin.

    Science.gov (United States)

    Rajendran, Madhusudan; Nachbagauer, Raffael; Ermler, Megan E; Bunduc, Paul; Amanat, Fatima; Izikson, Ruvim; Cox, Manon; Palese, Peter; Eichelberger, Maryna; Krammer, Florian

    2017-03-21

    Antibody responses to influenza virus hemagglutinin provide protection against infection and are well studied. Less is known about the human antibody responses to the second surface glycoprotein, neuraminidase. Here, we assessed human antibody reactivity to a panel of N1, N2, and influenza B virus neuraminidases in different age groups, including children, adults, and the elderly. Using enzyme-linked immunosorbent assays (ELISA), we determined the breadth, magnitude, and isotype distribution of neuraminidase antibody responses to historic, current, and avian strains, as well as to recent isolates to which these individuals have not been exposed. It appears that antibody levels against N1 neuraminidases were lower than those against N2 or B neuraminidases. The anti-neuraminidase antibody levels increased with age and were, in general, highest against strains that circulated during the childhood of the tested individuals, providing evidence for "original antigenic sin." Titers measured by ELISA correlated well with titers measured by the neuraminidase inhibition assays. However, in the case of the 2009 pandemic H1N1 virus, we found evidence of interference from antibodies binding to the conserved stalk domain of the hemagglutinin. In conclusion, we found that antibodies against the neuraminidase differ in magnitude and breadth between subtypes and age groups in the human population. (This study has been registered at ClinicalTrials.gov under registration no. NCT00336453, NCT00539981, and NCT00395174.)IMPORTANCE Anti-neuraminidase antibodies can afford broad protection from influenza virus infection in animal models and humans. However, little is known about the breadth and magnitude of the anti-neuraminidase response in the human population. Here we assessed antibody levels of children, adults, and the elderly against a panel of N1, N2, and type B influenza virus neuraminidases. We demonstrated that antibody levels measured by ELISA correlate well with functional

  8. Fc gamma receptor IIIB-NA gene frequencies in patients with systemic lupus erythematosus and healthy individuals of Malay and Chinese ethnicity.

    Science.gov (United States)

    Yap, S N; Phipps, M E; Manivasagar, M; Bosco, J J

    1999-06-01

    The neutrophil antigen (NA)1 and 2 is coded by two recognized allelic forms of Fc gamma receptor IIIB (FcgammaRIIIB). FcgammaRIIIb is a low affinity receptor and preferentially removes immune complexes from the circulation. Systemic lupus erythematosus (SLE) is an autoimmune and polygenic disorder characterized by accumulation of autoimmune complexes. The majority of SLE patients in our medical center are of Chinese ethnicity, followed by Malay and Indian. Recently, studies have focussed on the Fc receptors in different ethnic groups and their relation to SLE. We chose to study the gene distribution of this receptor in the Chinese and Malays population in Malaysia. We designed a polymerase chain reaction allele specific primers (PCR-ASP) method to distinguish the two allelic forms. Genomic DNA was isolated from the peripheral blood of 183 Chinese and 55 Malays SLE patients as well as 100 Chinese and 50 Malays healthy controls. Genotyping of Chinese SLE patients revealed that the gene frequencies for FcgammaRIIIB-NA1 and FcgammaRIIIB-NA2 were 0.648 and 0.347, while in the ethnically matched healthy controls they were 0.68 and 0.32, respectively. One out of the 183 Chinese SLE patients was identified as a NA-null due to the absence of PCR product for both alleles. The FcgammaRIIIB-NA1 and FcgammaRIIIB-NA2 allele frequencies for both the Malays SLE and healthy controls were 0.62 and 0.38.

  9. Baicalein, Ethyl Acetate, and Chloroform Extracts of Scutellaria baicalensis Inhibit the Neuraminidase Activity of Pandemic 2009 H1N1 and Seasonal Influenza A Viruses

    Directory of Open Access Journals (Sweden)

    Mann-Jen Hour

    2013-01-01

    Full Text Available This study rated antiviral activity of Scutellaria baicalensis Georgi (S. baicalensis extracts against influenza A virus subtypes, for example, pandemic 2009 H1N1, seasonal H1N1 and H3N2. Ethyl acetate (EtOAc and chloroform extracts inhibited in vitro neuraminidase (NA enzymatic activity and viral replication more than methanol (MeOH extract. EtOAc extract demonstrated NA inhibition IC50 values ranging from 73.16 to 487.40 μg/mL and plaque reduction IC50 values ranging from 23.7 to 27.4 μg/mL. Chloroform extract showed antiviral activities with plaque reduction IC50 values ranging from 14.16 to 41.49 μg/mL Time-of-addition assay indicated that EtOAc and chloroform extracts also significantly inhibited virus yields after infection. HPLC analysis demonstrated that baicalin was dominant in the MeOH extract; baicalein and chrysin were rich in the EtOAc and chloroform extracts. Molecular simulation revealed baicalein hydrogen bonding with Glu277 as well as hydrophobic and Van der Waals interactions with Ile222, Arg224, Ser246, and Tyr347 in NA1 active sites of NA1. Baicalein inhibited in vitro replication of influenza A viruses pandemic 2009 H1N1 (IC50 = 0.018 μM and seasonal 2007 H1N1 using plaque reduction assays. A combination of low-dose baicalein with other anti-influenza agents could be applicable for development of alternative remedies treating influenza A virus infection.

  10. Molecular basis for broad neuraminidase immunity: conserved epitopes in seasonal and pandemic H1N1 as well as H5N1 influenza viruses.

    Science.gov (United States)

    Wan, Hongquan; Gao, Jin; Xu, Kemin; Chen, Hongjun; Couzens, Laura K; Rivers, Katie H; Easterbrook, Judy D; Yang, Kevin; Zhong, Lei; Rajabi, Mohsen; Ye, Jianqiang; Sultana, Ishrat; Wan, Xiu-Feng; Liu, Xiufan; Perez, Daniel R; Taubenberger, Jeffery K; Eichelberger, Maryna C

    2013-08-01

    Influenza A viruses, including H1N1 and H5N1 subtypes, pose a serious threat to public health. Neuraminidase (NA)-related immunity contributes to protection against influenza virus infection. Antibodies to the N1 subtype provide protection against homologous and heterologous H1N1 as well as H5N1 virus challenge. Since neither the strain-specific nor conserved epitopes of N1 have been identified, we generated a panel of mouse monoclonal antibodies (MAbs) that exhibit different reactivity spectra with H1N1 and H5N1 viruses and used these MAbs to map N1 antigenic domains. We identified 12 amino acids essential for MAb binding to the NA of a recent seasonal H1N1 virus, A/Brisbane/59/2007. Of these, residues 248, 249, 250, 341, and 343 are recognized by strain-specific group A MAbs, while residues 273, 338, and 339 are within conserved epitope(s), which allows cross-reactive group B MAbs to bind the NAs of seasonal H1N1 and the 1918 and 2009 pandemic (09pdm) H1N1 as well as H5N1 viruses. A single dose of group B MAbs administered prophylactically fully protected mice against lethal challenge with seasonal and 09pdm H1N1 viruses and resulted in significant protection against the highly pathogenic wild-type H5N1 virus. Another three N1 residues (at positions 396, 397, and 456) are essential for binding of cross-reactive group E MAbs, which differ from group B MAbs in that they do not bind 09pdm H1N1 viruses. The identification of conserved N1 epitopes reveals the molecular basis for NA-mediated immunity between H1N1 and H5N1 viruses and demonstrates the potential for developing broadly protective NA-specific antibody treatments for influenza.

  11. Estudo da metilação nos genes IFN-y e IL-10 na doença periodontal

    OpenAIRE

    Michelle Beatriz Viana

    2009-01-01

    A metilação do DNA é um fator epigenético caracterizado pela adição de um grupo metil em citosinas de regiões CpG, promovendo modificações na estrutura do DNA sem envolver alterações na seqüência de nucleotídeos. A presença de metilação na região promotora dos genes pode provocar o silenciamento transcricional dos mesmos. Alterações epigenéticas podem ocorrer em genes de citocinas afetando a expressão dessas moléculas envolvidas no processo inflamatório. Estudos recentes apontam para a partic...

  12. Differentially expressed genes in Populus simonii x P. nigra in respnse to NaCl stress using cDNA-AFLP

    Science.gov (United States)

    Salinity is an important environmental factor limiting growth and productivity of plants, and affects almost every aspect of the plant physiology and biochemistry. The objective of this study was to apply cDNA-AFLP and to identify differentially expressed genes in response to NaCl stress vs. no-stre...

  13. C-Methylated Flavonoids from Cleistocalyx operculatus and Their Inhibitory Effects on Novel Influenza A (H1N1) Neuraminidase

    DEFF Research Database (Denmark)

    Dao, Trong-Tuan; Tung, Bui-Thanh; Nguyen, Phi-Hung

    2010-01-01

    As part of an ongoing study focused on the discovery of anti-influenza agents from plants, four new (1-4) and 10 known (5-14) C-methylated flavonoids were isolated from a methanol extract of Cleistocalyx operculatus buds using an influenza H1N1 neuraminidase inhibition assay. Compounds 4, 7, 8...

  14. Correlation of haemagglutinin-neuraminidase and fusion protein content with protective antibody response after immunisation with inactivated Newcastle disease vaccines.

    NARCIS (Netherlands)

    Maas, R.A.; Komen, M.; Diepen, van M.; Oei, H.L.; Claassen, I.J.T.M.

    2003-01-01

    The correlation between the antigen content of inactivated Newcastle disease (ND) oil emulsion-vaccines and the serological response after immunisation was studied. The haemagglutinin-neuraminidase (HN) and fusion (F) proteins of Newcastle disease virus (NDV) were quantified in 33 inactivated

  15. Financial competing interests were associated with favorable conclusions and greater author productivity in nonsystematic reviews of neuraminidase inhibitors.

    Science.gov (United States)

    Dunn, Adam G; Zhou, Xujuan; Hudgins, Joel; Arachi, Diana; Mandl, Kenneth D; Coiera, Enrico; Bourgeois, Florence T

    2016-12-01

    To characterize the conclusions and production of nonsystematic reviews about neuraminidase inhibitors relative to financial competing interests held by the authors. We searched for articles about neuraminidase inhibitors and influenza (January 2005 to April 2015), identifying nonsystematic reviews and grading them according to the favorable/nonfavorable presentation of evidence on safety and efficacy. We recorded financial competing interests disclosed in the reviews and from other articles written by their authors. We measured associations between competing interests, author productivity, and conclusions. Among 213 nonsystematic reviews, 138 (65%) presented favorable conclusions. Financial competing interests were identified for 26% (137/532) of authors; 51% (108/213) of reviews were associated with a financial competing interest. Reviews produced exclusively by authors with financial competing interests (33%; 71/213) were more likely to present favorable conclusions than reviews with no competing interests (risk ratio 1.27; 95% confidence interval 1.03-1.55). Authors with financial competing interests published more articles about neuraminidase inhibitors than their counterparts. Half of nonsystematic reviews about neuraminidase inhibitors included an author with a financial competing interest. Reviews produced exclusively by these authors were more likely to present favorable conclusions, and authors with financial competing interests published a greater number of reviews. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Naïve Induced Pluripotent Stem Cells Generated From β-Thalassemia Fibroblasts Allow Efficient Gene Correction With CRISPR/Cas9.

    Science.gov (United States)

    Yang, Yuanyuan; Zhang, Xiaobai; Yi, Li; Hou, Zhenzhen; Chen, Jiayu; Kou, Xiaochen; Zhao, Yanhong; Wang, Hong; Sun, Xiao-Fang; Jiang, Cizhong; Wang, Yixuan; Gao, Shaorong

    2016-01-01

    Conventional primed human embryonic stem cells and induced pluripotent stem cells (iPSCs) exhibit molecular and biological characteristics distinct from pluripotent stem cells in the naïve state. Although naïve pluripotent stem cells show much higher levels of self-renewal ability and multidifferentiation capacity, it is unknown whether naïve iPSCs can be generated directly from patient somatic cells and will be superior to primed iPSCs. In the present study, we used an established 5i/L/FA system to directly reprogram fibroblasts of a patient with β-thalassemia into transgene-free naïve iPSCs with molecular signatures of ground-state pluripotency. Furthermore, these naïve iPSCs can efficiently produce cross-species chimeras. Importantly, using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease genome editing system, these naïve iPSCs exhibit significantly improved gene-correction efficiencies compared with the corresponding primed iPSCs. Furthermore, human naïve iPSCs could be directly generated from noninvasively collected urinary cells, which are easily acquired and thus represent an excellent cell resource for further clinical trials. Therefore, our findings demonstrate the feasibility and superiority of using patient-specific iPSCs in the naïve state for disease modeling, gene editing, and future clinical therapy. In the present study, transgene-free naïve induced pluripotent stem cells (iPSCs) directly converted from the fibroblasts of a patient with β-thalassemia in a defined culture system were generated. These naïve iPSCs, which show ground-state pluripotency, exhibited significantly improved single-cell cloning ability, recovery capacity, and gene-targeting efficiency compared with conventional primed iPSCs. These results provide an improved strategy for personalized treatment of genetic diseases such as β-thalassemia. ©AlphaMed Press.

  17. Expression of the sigma35 and cry2AB genes involved in Bacillus thuringiensis virulence Expressão dos genes sigma35 e cry2AB envolvidos na virulência de Bacillus thuringiensis

    Directory of Open Access Journals (Sweden)

    Ana Maria Guidelli-Thuler

    2009-06-01

    Full Text Available There are several genes involved in Bacillus thuringiensis sporulation. The regulation and expression of these genes results in an upregulation in Cry protein production, and this is responsible for the death of insect larvae infected by Bacillus thuringiensis. Gene expression was monitored in Bacillus thuringiensis during three developmental phases. DNA macroarrays were constructed for selected genes whose sequences are available in the GenBank database. These genes were hybridized to cDNA sequences from B. thuringiensis var. kurstaki HD-1. cDNA probes were synthesized by reverse transcription from B. thuringiensis RNA templates extracted during the exponential (log growth, stationary and sporulation phases, and labeled with 33PadCTP. Two genes were differentially expressed levels during the different developmental phases. One of these genes is related to sigma factor (sigma35, and the other is a cry gene (cry2Ab. There were differences between the differential levels of expression of various genes and among the expression detected for different combinations of the sigma factor and cry2Ab genes. The maximum difference in expression was observed for the gene encoding sigma35 factor in the log phase, which was also expressed at a high level during the sporulation phase. The cry2Ab gene was only expressed at a high level in the log phase, but at very low levels in the other phases when compared to the sigma35.Muitos genes estão envolvidos nos mecanismos de esporulação da bactéria Bacillus thuringiensis. A regulação e expressão desses genes resultam em uma produção massiva da proteína Cry, responsável pela morte das larvas de muitos insetos. Neste trabalho monitorou-se a expressão de genes de Bacillus thuringiensis, ao longo de três fases de seu desenvolvimento. Foram construídos macroarrays de DNA dos genes selecionados, cujas seqüências estão disponibilizadas no GenBank. Estes genes foram hibridizados com cDNAs obtidos de B

  18. Effect of verapamil and lidocaine on TRPM and NaV1.9 gene expressions in renal ischemia-reperfusion.

    Science.gov (United States)

    Dusmez, D; Cengiz, B; Yumrutas, O; Demir, T; Oztuzcu, S; Demiryurek, S; Tutar, E; Bayraktar, R; Bulut, A; Simsek, H; Daglı, S Nur; Kılıc, T; Bagcı, C

    2014-01-01

    To determine effects on calcium and sodium channels of Ca(2+) and Na(+) channel blockers in the present study, expression levels of TRPM1, TRPM2, TRPM3, TRPM4, TRPM5, TRPM6, TRPM7, TRPM8, and NaV1.9 genes were evaluated in kidney tissues after induced ischemia-reperfusion. Forty albino Wistar male rats were equally divided into 4 groups as follows: group I: control group (n = 10), group II: ischemia group (60 minutes of ischemia + 48 hours of reperfusion; n = 10), group III: ischemia (60 minutes of ischemia + 48 hours of reperfusion) + calcium channel blocker (n = 8), group IV: ischemia (60 minutes of ischemia + 48 hours of reperfusion) + sodium channel blocker (n = 8). When compared to ischemia group expression levels of TRPM2, TRPM4, TRPM6, and NaV1.9 in Ca(2+) and Na(+) channel blocker groups were increased, whereas that of TRPM7 was decreased. However, expression levels of TRPM1, TRPM3, TRPM5, and TRPM8 were not determined in kidney tissue. Histologically, the Ca(2+) channel blocker verapamil and the Na(+) channel blocker lidocaine inhibited the cell death in kidney tissue compared to control. Our study suggested that verapamil and lidocaine significantly reduce the degree of ischemia-reperfusion injury due to effects to TRPM and Nav1.9 genes. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Biological Characterizations of H5Nx Avian Influenza Viruses Embodying Different Neuraminidases

    Directory of Open Access Journals (Sweden)

    Yuandi Yu

    2017-06-01

    Full Text Available The H5 subtype virus of Highly Pathogenic Avian Influenza Virus has caused huge economic losses to the poultry industry and is a threat to human health. Until 2010, H5N1 subtype virus was the major genotype in China. Since 2011, reassortant H5N2, H5N6, and H5N8 viruses were identified in domestic poultry in China. The clade 2.3.4.4 H5N6 and H5N8 AIV has now spread to most of China. Clade 2.3.4.4 H5N6 virus has caused 17 human deaths. However, the prevalence, pathogenicity, and transmissibility of the distinct NA reassortment with H5 subtypes viruses (H5Nx is unknown. We constructed five clade 2.3.4.4 reassortant H5Nx viruses that shared the same HA and six internal gene segments. The NA gene segment was replaced with N1, N2, N6, ΔN6 (with an 11 amino acid deletion at the 58th to 68th of NA stalk region, and N8 strains, respectively. The reassortant viruses with distinct NAs of clade 2.3.4.4 H5 subtype had different degrees of fitness. All reassortant H5Nx viruses formed plaques on MDCK cell monolayers, but the ΔH5N6 grew more efficiently in mammalian and avian cells. The reassortant H5Nx viruses were more virulent in mice as compared to the H5N2 virus. The H5N6 and H5N8 reassortant viruses exhibited enhanced pathogenicity and transmissibility in chickens as compared to the H5N1 reassortant virus. We suggest that comprehensive surveillance work should be undertaken to monitor the H5Nx viruses.

  20. Molecular analysis of the Na+/H+ exchanger gene family and its role in salt stress in Medicago truncatula

    Science.gov (United States)

    High salinity in irrigation water is detrimental to plant growth and productivity. Plants develop various mechanisms and strategies to cope with salinity. One important mechanism is keeping the cytosolic Na+ concentration low by sequestering Na+ in vacuoles, a process facilitated by Na+/H+ exchanger...

  1. Evaluation of the absolute affinity of neuraminidase inhibitor using steered molecular dynamics simulations.

    Science.gov (United States)

    Tam, Nguyen Minh; Nguyen, Minh Tho; Ngo, Son Tung

    2017-08-24

    The absolute free energy difference of binding (ΔG) between neuraminidase and its inhibitor was evaluated using fast pulling of ligand (FPL) method over steered molecular dynamics (SMD) simulations. The metric was computed through linear interaction approximation. Binding nature was described by free energy differences of electrostatic and van der Waals (vdW) interactions. The finding indicates that vdW metric is dominant over electrostatics in binding process. The computed values are in good agreement with experimental data with a correlation coefficient of R=0.82 and error of σΔGexp=2.2kcal/mol. The results were observed using Amber99SB-ILDN force field in comparison with CHARMM27 and GROMOS96 43a1 force fields. Obtained results may stimulate the search for an Influenza therapy. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Virtual screening of Indonesian flavonoid as neuraminidase inhibitor of influenza a subtype H5N1

    Science.gov (United States)

    Parikesit, A. A.; Ardiansah, B.; Handayani, D. M.; Tambunan, U. S. F.; Kerami, D.

    2016-02-01

    Highly Pathogenic Avian Influenza (HPAI) H5N1 poses a significant threat to animal and human health worldwide. The number of H5N1 infection in Indonesia is the highest during 2005-2013, with a mortality rate up to 83%. A mutation that occurred in H5N1 strain made it resistant to commercial antiviral agents such as oseltamivir and zanamivir, so the more potent antiviral agent is needed. In this study, virtual screening of Indonesian flavonoid as neuraminidase inhibitor of H5N1 was conducted. Total 491 flavonoid compound obtained from HerbalDB were screened. Molecular docking was performed using MOE 2008.10. This research resulted in Guajavin B as the best ligand.

  3. The SbSOS1 gene from the extreme halophyte Salicornia brachiata enhances Na+ loading in xylem and confers salt tolerance in transgenic tobacco

    Directory of Open Access Journals (Sweden)

    Yadav Narendra

    2012-10-01

    Full Text Available Abstract Background Soil salinity adversely affects plant growth and development and disturbs intracellular ion homeostasis resulting cellular toxicity. The Salt Overly Sensitive 1 (SOS1 gene encodes a plasma membrane Na+/H+ antiporter that plays an important role in imparting salt stress tolerance to plants. Here, we report the cloning and characterisation of the SbSOS1 gene from Salicornia brachiata, an extreme halophyte. Results The SbSOS1 gene is 3774 bp long and encodes a protein of 1159 amino acids. SbSOS1 exhibited a greater level of constitutive expression in roots than in shoots and was further increased by salt stress. Overexpressing the S. brachiata SbSOS1 gene in tobacco conferred high salt tolerance, promoted seed germination and increased root length, shoot length, leaf area, fresh weight, dry weight, relative water content (RWC, chlorophyll, K+/Na+ ratio, membrane stability index, soluble sugar, proline and amino acid content relative to wild type (WT plants. Transgenic plants exhibited reductions in electrolyte leakage, reactive oxygen species (ROS and MDA content in response to salt stress, which probably occurred because of reduced cytosolic Na+ content and oxidative damage. At higher salt stress, transgenic tobacco plants exhibited reduced Na+ content in root and leaf and higher concentrations in stem and xylem sap relative to WT, which suggests a role of SbSOS1 in Na+ loading to xylem from root and leaf tissues. Transgenic lines also showed increased K+ and Ca2+ content in root tissue compared to WT, which reflect that SbSOS1 indirectly affects the other transporters activity. Conclusions Overexpression of SbSOS1 in tobacco conferred a high degree of salt tolerance, enhanced plant growth and altered physiological and biochemical parameters in response to salt stress. In addition to Na+ efflux outside the plasma membrane, SbSOS1 also helps to maintain variable Na+ content in different organs and also affect the other

  4. Molecular Characterizations of Surface Proteins Hemagglutinin and Neuraminidase from Recent H5Nx Avian Influenza Viruses

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Hua; Carney, Paul J.; Mishin, Vasiliy P.; Guo, Zhu; Chang, Jessie C.; Wentworth, David E.; Gubareva, Larisa V.; Stevens, James; Schultz-Cherry, S.

    2016-04-06

    ABSTRACT

    During 2014, a subclade 2.3.4.4 highly pathogenic avian influenza (HPAI) A(H5N8) virus caused poultry outbreaks around the world. In late 2014/early 2015, the virus was detected in wild birds in Canada and the United States, and these viruses also gave rise to reassortant progeny, composed of viral RNA segments (vRNAs) from both Eurasian and North American lineages. In particular, viruses were found with N1, N2, and N8 neuraminidase vRNAs, and these are collectively referred to as H5Nx viruses. In the United States, more than 48 million domestic birds have been affected. Here we present a detailed structural and biochemical analysis of the surface antigens of H5N1, H5N2, and H5N8 viruses in addition to those of a recent human H5N6 virus. Our results with recombinant hemagglutinin reveal that these viruses have a strict avian receptor binding preference, while recombinantly expressed neuraminidases are sensitive to FDA-approved and investigational antivirals. Although H5Nx viruses currently pose a low risk to humans, it is important to maintain surveillance of these circulating viruses and to continually assess future changes that may increase their pandemic potential.

    IMPORTANCEThe H5Nx viruses emerging in North America, Europe, and Asia pose a great public health concern. Here we report a molecular and structural study of the major surface proteins of several H5Nx influenza viruses. Our results improve the understanding of these new viruses and provide important information on their receptor preferences and susceptibilities to antivirals, which are central to pandemic risk assessment.

  5. Integrin-mediated cell migration is blocked by inhibitors of human neuraminidase.

    Science.gov (United States)

    Jia, Feng; Howlader, Md Amran; Cairo, Christopher W

    2016-09-01

    Integrins are critical receptors in cell migration and adhesion. A number of mechanisms are known to regulate the function of integrins, including phosphorylation, conformational change, and cytoskeletal anchoring. We investigated whether native neuraminidase (Neu, or sialidase) enzymes which modify glycolipids could play a role in regulating integrin-mediated cell migration. Using a scratch assay, we found that exogenously added Neu3 and Neu4 activity altered rates of cell migration. We observed that Neu4 increased the rate of migration in two cell lines (HeLa, A549); while Neu3 only increased migration in HeLa cells. A bacterial neuraminidase was able to increase the rate of migration in HeLa, but not in A549 cells. Treatment of cells with complex gangliosides (GM1, GD1a, GD1b, and GT1b) resulted in decreased cell migration rates, while LacCer was able to increase rates of migration in both lines. Importantly, our results show that treatment of cells with inhibitors of native Neu enzymes had a dramatic effect on the rates of cell migration. The most potent compound tested targeted the human Neu4 isoenzyme, and was able to substantially reduce the rate of cell migration. We found that the lateral mobility of integrins was reduced by treatment of cells with Neu3, suggesting that Neu3 enzyme activity resulted in changes to integrin-co-receptor or integrin-cytoskeleton interactions. Finally, our results support the hypothesis that inhibitors of human Neu can be used to investigate mechanisms of cell migration and for the development of anti-adhesive therapies. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. [Inhibitory effect of NaV1.9 gene silencing on proliferation, phagocytosis and migration in RAW264.7 cells].

    Science.gov (United States)

    Luo, Yuechen; Zhou, Xin; Ji, Wenjie; Sun, Haiying; Lu, Ruiyi; Jiang, Tiemin; Li, Yuming

    2013-03-01

    To establish the cell line with stable voltage-gated sodium channels (VGSCs/NaVs) α subunit NaV1.9 gene silencing through RNA interference (RNAi) in murine RAW264.7 macrophages, and to investigate proliferation, phagocytosis and migration in this cell line. The stable NaV1.9-deficient cell line was generated by selection in G418 after the transfection of short hairpin (shRNA) plasmid with Lipofectamine TM2000. RNAi efficiency was qualified by RT-PCR; proliferation ability was measured by CCK-8 assay; cell cycle and phagocytic ability were analyzed by flow cytometry; and migrating ability was detected by Transwell migration assay. Stable NaV1.9-deficient cell line was established and the expression of NaV1.9 was reduced by 80%. CCK-8 assay and flow cytometry showed that the proliferation of the NaV1.9-deficient cell line was inhibited (PNaV1.9-deficient cells we successfully constructed, proliferation, phagocytosis and migration were obviously inhibited.

  7. Neuraminidase Activity in Streptococcus sanguis and in the Viridans Group, and Occurrence of Acylneuraminate Lyase in Viridans Organisms Isolated from Patients with Septicemia

    Science.gov (United States)

    Müller, H. E.

    1974-01-01

    The enzyme neuraminidase (EC 3.2.1.18) was found to be strongly active in different types of Streptococcus sanguis and S. viridans, and, in addition, the occurrence of the enzyme acylneuraminate pyruvate lyase (EC 4.1.3.3) was described in S. viridans. The enzyme-active bacteria strains were isolated from blood cultures of patients with septicemia. Whereas S. sanguis lost its strong neuraminidase activity after some weeks, S. viridans retained its enzyme activity for a long time in culture. Immunoelectrophoretic studies of the blood cultures of patients with streptococcal infections showed the loss of neuraminic acid in most glycoproteins of the serum, proving the in vivo action of neuraminidase. The pathogenic role of neuraminidase is discussed in streptococcal septicemia from the viewpoint of present knowledge. Images PMID:4816461

  8. Sodium-iodine symporter gene expression controlled by the EGR-1 promoter: biodistribution, imaging and in vitro radionuclide therapy with Na(131)I.

    Science.gov (United States)

    Tang, Jun; Wang, Xiaoxia; Xu, Yuanqi; Shi, Yizhen; Liu, Zengli; Yang, Yi

    2015-02-01

    The objective of this study is to explore the feasibility of radioiodine treatment for cervical cancer using the early growth response (Egr-1) promoter to control sodium-iodine symporter (hNIS) gene expression. The hNIS gene was previously transfected into Hela cells under the control of either the cytomegalovirus (CMV) or Egr-1 promoters. Na(125)I uptake was measured in the presence or absence of NaClO4. Na(125)I efflux was measured. The effects of external beam radiation on iodine uptake and retention were studied. The cytotoxic effects of (131)I were measured by clonogenic assay. The Na(125)I biodistribution was obtained using mice bearing control and transfected cells. The %ID/g of tumor and major organs were obtained for a range of times up to 48 hours post injection and the ratio of tumor to non-tumor activity (T/NT) was calculated. Tumors were imaged with Na(131)I and (99m)TcO4 (-), and the ratio of tumor to background activity (T/B) was calculated. Na(125)I uptake in Hela cells was minimal in the absence of hNIS. Uptake in the transfected cells was strong, and could be blocked by NaClO4. The iodine uptake of Hela-Egr-1-hNIS cells increased after the irradiation, and the magnitude of this effect approximately matched the radiation dose delivered. The efflux of 125I was affected by neither the promoter sequence nor pre-irradiation. (131)I reduced the clonogenic survival of symporter expressing cells, relative to the parental line. The effect was greatest in cells where hNIS was driven by the CMV promoter. Tumors formed from Hela-Egr-1-hNIS concentrated Na(125)I over a 12 hour period, in contrast to untransfected cells. These tumors could also be successfully imaged using either Na(131)I or (99m)TcO4 (-). (131)I uptake peaked at 4h, while (99m)TcO4 (-) accumulated over approximately 20 hours. In vivo uptake of (131)I and (99m)TcO4 (-) was slightly higher in cells transfected with the Egr-1 promoter, compared to CMV. Hela-Egr-1-hNIS cells demonstrate highly

  9. Fc gamma RIIIB gene duplication: evidence for presence and expression of three distinct Fc gamma RIIIB genes in NA(1+,2+)SH(+) individuals

    NARCIS (Netherlands)

    Koene, H. R.; Kleijer, M.; Roos, D.; de Haas, M.; von dem Borne, A. E.

    1998-01-01

    Recently, a new alloantigen on IgG Fc receptor type IIIb (Fc gamma RIIIb), SH, was described (Bux et al, Blood 89:1027, 1997). We identified three healthy individuals whose neutrophils reacted positively with the SH antiserum. The neutrophil antigen (NA) phenotype of all three donors was NA(1+,2+).

  10. Adamantane and Neuraminidase resistant influenza A/H3N2 isolated in Iran from 2005 to 2013

    Directory of Open Access Journals (Sweden)

    Jila Yavarian

    2014-04-01

    Conclusion: This study showed circulating A/H3N2 viruses was resistant to adaman-tanes but susceptible to neuraminidase inhibitors. The national data analyzed in this re-search may help increase knowledge about influenza virus antiviral drug resistance, which is a global public health concern. The authors suggested continuing this study and also the investigation of antiviral drug resistance of influenza A/H1N1 and B viruses.

  11. Prognosis of hospitalized patients with 2009 H1N1 influenza in Spain: influence of neuraminidase inhibitors

    Science.gov (United States)

    Delgado-Rodríguez, Miguel; Castilla, Jesús; Godoy, Pere; Martín, Vicente; Soldevila, Nuria; Alonso, Jordi; Astray, Jenaro; Baricot, Maretva; Cantón, Rafael; Castro, Ady; Gónzález-Candelas, Fernando; Mayoral, José María; Quintana, José María; Pumarola, Tomás; Tamames, Sonia; Sáez, Marc; Domínguez, Angela

    2012-01-01

    Background The H1N1 influenza pandemic strain has been associated with a poor prognosis in hospitalized patients. The present report evaluates the factors influencing prognosis. Methods A total of 813 patients hospitalized with H1N1 influenza in 36 hospitals (nationwide) in Spain were analysed. Detailed histories of variables preceding hospital admission were obtained by interview, validating data on medications and vaccine with their attending physicians. Data on treatment and complications during hospital stay were recorded. As definition of poor outcome, the endpoints of death and admission to intensive care were combined; and as a further outcome, length of stay was used. Results The mean age was 38.5 years (SD 22.8 years). There were 10 deaths and 79 admissions to intensive care (combined, 88). The use of neuraminidase inhibitors was reported by 495 patients (60.9%). The variables significantly associated with a poor outcome were diabetes (OR = 2.21, 95% CI = 1.21–4.02), corticosteroid therapy (OR = 3.37, 95% CI = 1.39–8.20) and use of histamine-2 receptor antagonists (OR = 2.68, 95% CI = 1.14–6.36), while the use of neuraminidase inhibitors (OR = 0.57, 95% CI = 0.34–0.94) was protective. Neuraminidase inhibitors within the first 2 days after the influenza onset reduced hospital stay by a mean of 1.9 days (95% CI = 4.7–6.6). Conclusions The use of neuraminidase inhibitors decreases the length of hospital stay and admission to intensive care and/or death. PMID:22467633

  12. Assessment of Streptococcus pneumoniae Capsule in Conjunctivitis and Keratitis in vivo Neuraminidase Activity Increases in Nonencapsulated Pneumococci following Conjunctival Infection

    Science.gov (United States)

    Norcross, Erin W.; Tullos, Nathan A.; Taylor, Sidney D.; Sanders, Melissa E.; Marquart, Mary E.

    2010-01-01

    Purpose The pneumococcal capsule is required for pathogenesis in systemic infections, yet reports show most conjunctivitis outbreaks are caused by nonencapsulated pneumococci, while keratitis infections are caused by encapsulated strains. This study aims to determine the effect of capsule in pneumococcal keratitis and conjunctivitis in rabbit models of infection. Methods A capsule-deficient isogenic mutant was created using homologous transformation. Parent and mutant strains were injected within the upper bulbar conjunctiva (conjunctivitis) or into the corneal stroma (keratitis) of New Zealand white rabbits. Clinical examinations were performed 24 and 48 hr post-infection at which time corneas or conjunctivae were removed, homogenized, and plated to determine the recovered bacterial load. Whole eyes were removed for histological examination. The neuraminidase activity was determined following in vitro and in vivo growth. Results There were no significant differences in clinical scores between the eyes infected with the parent or mutant for either infection, nor was there a difference in the amount of bacteria recovered from the cornea. In the conjunctivae, however, the mutant strain was cleared by the host faster than the parent strain. Histological examination showed slightly more infiltrating polymorphonuclear leukocytes (PMN) and macrophages in the conjunctivae infected with the parent strain. The neuraminidase activity of both strains was not significantly different when the strains were grown in vitro. However, the neuraminidase activity of the parent was significantly less than that of the mutant at 3 and 12 hr post conjunctival infection. Conclusions Although more outbreaks of pneumococcal conjunctivitis are tied to nonencapsulated S. pneumoniae strains, this study showed that an encapsulated strain was capable of establishing conjunctivitis in a rabbit injection model and survive attack by the host immune system longer than its nonencapsulated isogenic

  13. Kinetic, thermodynamic and structural analysis of tamiphosphor binding to neuraminidase of H1N1 (2009) pandemic influenza

    Czech Academy of Sciences Publication Activity Database

    Albinana, C. B.; Machara, A.; Řezáčová, Pavlína; Pachl, Petr; Konvalinka, Jan; Kožíšek, Milan

    2016-01-01

    Roč. 121, Oct 4 (2016), s. 100-109 ISSN 0223-5234 R&D Projects: GA ČR GA13-19561S; GA MŠk LO1302; GA MŠk(CZ) LO1304 Institutional support: RVO:61388963 Keywords : influenza neuraminidase * oseltamivir * tamiphosphor * isothermal titration calorimetry * crystal structure * lattice-translocation defect Subject RIV: CE - Biochemistry Impact factor: 4.519, year: 2016

  14. O estroma da medula ossea e a sua influencia na expressão de genes de resistencia e sensibilidade a quimioterapicos na leucemia linfoide aguda (LLA) pediatrica

    OpenAIRE

    Angelo Brunelli Albertoni Laranjeira

    2007-01-01

    Resumo: A resistência intrínseca ou adquirida aos compostos quimioterápicos é uma das mais importantes causas dos insucessos no tratamento das LLAs pediátricas. A interação da LLA com o microambiente da medula óssea contribui para a proliferação e resistência ao regime quimioterápico das células leucêmicas através de uma grande variedade de mecanismos celulares que provavelmente incluem: aumento da expressão de transportadores celulares, aumento no processo de reparo do DNA, diminuição na reg...

  15. Sequence analysis of haemagglutinin and neuraminidase of H1N1 strain from a patient coinfected with Mycobacterium tuberculosis.

    Science.gov (United States)

    Alghamdi, Ahmed N; Mahfouz, Mohammad E; Hamdi, Fahd A; Al Aboud, Daifullah; Al-Laylah, Tawfiq Z; Alotaibi, Mohammed I; Al-Thomali, Khalid W A; Abdel-Moneim, Ahmed S

    2017-08-01

    The 2009 H1N1 pandemic (H1N1pdm09) was associated with a considerable influenza-related morbidity and mortality. Among the complications, Mycobacterial tuberculosis was recorded as a coinfection with influenza in rare cases. The full-length sequences of the viral haemagglutinin and neuraminidase of H1N1pdm09 influenza A virus were analyzed from a recently infected patient. The patient was chronically infected with Mycobacterium tuberculosis. Molecular modelling and in-silico docking of the virus, and other selected strains with the drug oseltamivir were conducted and compared. Sequence analysis of the viral haemagglutinin revealed it to be closely related to the 6B.1 clade, with high identity to the circulating H1N1pdm09 strains, and confirmed that the virus still harbouring high affinity to the α-2,6-sialic acid human receptor. The viral neuraminidase showed high identity to the neuraminidase of the recently circulating strains of the virus with no evidence of the development of oseltamivir-resistant mutants. Regular monitoring of the circulating strains is recommended to screen for a possible emergence of drug-resistant strains. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Ependymal denudation, aqueductal obliteration and hydrocephalus after a single injection of neuraminidase into the lateral ventricle of adult rats.

    Science.gov (United States)

    Grondona, J M; Pérez-Martín, M; Cifuentes, M; Pérez, J; Jiménez, A J; Pérez-Fígares, J M; Fernández-Llebrez, P

    1996-09-01

    To investigate the role of sialic acid in the ependyma of the rat brain, we injected neuraminidase from Clostridium perfingens into the lateral ventricle of 86 adult rats that were sacrificed at various time intervals. After administration of 10 micrograms neuraminidase, ciliated cuboidal ependymal cells of the lateral ventricles, third ventricle, cerebral aqueduct, and the rostral half of the fourth ventricle died and detached. The ependymal regions sealed by tight junctions such as the choroid plexus and the subcommissural organ were not affected. Debris was removed by infiltrating neutrophils and macrophagic cells. At the same time, after ependymal disappearance, the aqueduct was obliterated. In this region, mitoses were evident and cystic ependymal cells were frequent. Hydrocephalus of the lateral and third ventricles was evident 4 days after neuraminidase injection. Gliosis was restricted to the dorsal telencephalic wall of the injected lateral ventricle. It is thought that cleavage of sialic acid from ependymal surface glycoproteins or glycolipids, likely involved in cell adhesion, led to the detaching and death of the ependymal cells. Thereafter, ependymal loss, together with edema, led to fusion of the lateral walls of the cerebral aqueduct and this in turn provoked hydrocephalus of the third and lateral ventricles. This model of experimental hydrocephalus is compared with other models, in particular those of hydrocephalus after viral invasion of the cerebral ventricles.

  17. Expression of an alfalfa (Medicago sativa L.) peroxidase gene in transgenic Arabidopsis thaliana enhances resistance to NaCl and H2O2.

    Science.gov (United States)

    Teng, K; Xiao, G Z; Guo, W E; Yuan, J B; Li, J; Chao, Y H; Han, L B

    2016-05-23

    Peroxidases (PODs) are enzymes that play important roles in catalyzing the reduction of H2O2 and the oxidation of various substrates. They function in many different and important biological processes, such as defense mechanisms, immune responses, and pathogeny. The POD genes have been cloned and identified in many plants, but their function in alfalfa (Medicago sativa L.) is not known, to date. Based on the POD gene sequence (GenBank accession No. L36157.1), we cloned the POD gene in alfalfa, which was named MsPOD. MsPOD expression increased with increasing H2O2. The gene was expressed in all of the tissues, including the roots, stems, leaves, and flowers, particularly in stems and leaves under light/dark conditions. A subcellular analysis showed that MsPOD was localized outside the cells. Transgenic Arabidopsis with MsPOD exhibited increased resistance to H2O2 and NaCl. Moreover, POD activity in the transgenic plants was significantly higher than that in wild-type Arabidopsis. These results show that MsPOD plays an important role in resistance to H2O2 and NaCl.

  18. Electroporation-mediated in vivo gene delivery of the Na+/K+-ATPase pump reduced lung injury in a mouse model of lung contusion.

    Science.gov (United States)

    Machado-Aranda, David A; Suresh, M V; Yu, Bi; Raghavendran, Krishnan

    2012-01-01

    Lung contusion (LC) is an independent risk factor for acute respiratory distress syndrome. The final common pathway in ARDS involves accumulation of fluid in the alveoli. In this study, we demonstrate the application of a potential gene therapy approach by delivering the Na+/K+-ATPase pump subunits in a murine model of LC. We hypothesized that restoring the activity of the pump will result in removal of excess alveolar fluid and additionally reduce inflammation. Under anesthesia, C57/BL6 mice were struck along the right posterior axillary line 1 cm above the costal margin with a cortical contusion impactor. Immediately afterward, 100 μg of plasmid DNA coding for the α,β of the Na+/K+-ATPase pump were instilled into the lungs (LC-electroporation-pump group). Contusion only (LC-only) and a sham saline instillation group after contusion were used as controls (LC-electroporation-sham). By using a BTX 830 electroporator, eight electrical pulses of 200 V/cm field strength were applied transthoracically. Mice were killed at 24 hours, 48 hours, and 72 hours after delivery. Bronchial alveolar lavage was recollected to measure albumin and cytokines by enzyme-linked immunosorbent assay. Pulmonary compliance was measured, and lungs were subject to histopathologic analysis. After the electroporation and delivery of genes coding for the α,β subunits of the Na+/K+-ATPase pump, there was a significant mitigation of acute lung injury as evidenced by reduction in bronchial alveolar lavage levels of albumin, improved pressure volume curves, and reduced inflammation seen on histology. Electroporation-mediated gene transfer of the subunits of the Na+/K+-ATPase pump enhanced recovery from acute inflammatory lung injury after LC.

  19. Some novel insights into the binding of oseltamivir and zanamivir to H5N1 and N9 influenza virus neuraminidases:a homology modeling and flexible docking study

    Directory of Open Access Journals (Sweden)

    MARIJA L. MIHAJLOVIC

    2009-01-01

    Full Text Available In the context of the recent pandemic threat by the worldwide spread of H5N1 avian influenza, novel insights into the mechanism of ligand binding and interaction between various inhibitors (zanamivir – ZMV, oseltamivir – OTV, 2,3-didehydro-2-deoxy-N-acetylneuraminic acid – DANA, peramivir – PMV and neuraminidases (NA are of vital importance for the structure-based design of new anti-viral drugs. To address this issue, three-dimensional models of H5N1-NA and N9-NA were generated by homology modeling. Traditional residues within the active site throughout the family of NA protein structures were found to be highly conserved in H5N1-NA. A subtle variation between lipophilic and hydrophilic environments in H5N1-NA with respect to N9-NA was observed, thus shedding more light on the high resistance of some H5N1 strains to various NA inhibitors. Based on these models, an ArgusLab4/AScore flexible docking study was performed. The conformational differences between OTV bound to H5N1-NA and OTV bound to N9-NA were structurally identified and quantified. A slight difference of less than 1 kcal mol-1 between the OTV-N9 and OTV-N1 binding free energies is in agreement with the experimentally predicted free energy difference. The conformational differences between ZMV and OTV bound to either H5N1-NA or N9-NA were structurally identified. The binding free energies of the ZMV complexes, being slightly higher than those of OTV, are not in agreement with what was previously proposed using homology modeling. The differences between ZMV and OTV are suggested to be ascribed to the presence/absence of Asn166 in the active cavity of ZMV/OTV in H5N1-NA, and to the presence/absence of Ser165 in the binding site of ZMV/OTV in N9-NA. The charge distribution was evaluated using the semi-empirical AM1 method. The trends of the AM1 charges of the ZMV and OTV side chains in the complexes deviate from those previously reported.

  20. Gene Transcription and Virulence Potential of Listeria monocytogenes Strains After Exposure to Acidic and NaCl Stress

    DEFF Research Database (Denmark)

    Olesen, Inger; Vogensen, Finn Kvist; Jespersen, Lene

    2009-01-01

    transcription were observed both after exposure to shock (six genes) and after long-term adaptation to stress (18 genes). In the shock experiments, a transient induction of clpC and clpE was seen for both strains, while transient induction of sigB, inlA, and inlB was observed for strain 4140 only; actA was only...... response and invasion (clpC, clpP, inlA, inlB, prfA, and sigB) were induced in 4140. Long-term adaptation of EGD-e to NaCl stress increased transcription of genes important for the intracellular life cycle (actA, hly, iap, inlA, inlB, plcA, plcB, and prfA), while few changes were observed for 4140...

  1. Análise da expressão dos genes envolvidos na via de síntese de cafeína em plantas de café (Coffea arabica) apresentando diferentes teores de cafeína

    OpenAIRE

    Tavares, Aline Gomes

    2010-01-01

    A cafeína é um alcalóide do grupo das metilxantinas. Em café, sua síntese depende da ação de três enzimas: Metilxanthosina sintase (MS), Teobromina sintase (TS) e Cafeina sintase (CS). A crença de que o consumo de cafeína pode trazer problemas à saúde como também a sensibilidade aos efeitos da cafeína, em alguns consumidores, determinou um crescimento do mercado dos cafés descafeinados em pelo menos 10%. O processo de descafeinização hoje é baseado no uso de solventes, que acarreta em signifi...

  2. Variation in tissue Na(+) content and the activity of SOS1 genes among two species and two related genera of Chrysanthemum.

    Science.gov (United States)

    Gao, Jiaojiao; Sun, Jing; Cao, Peipei; Ren, Liping; Liu, Chen; Chen, Sumei; Chen, Fadi; Jiang, Jiafu

    2016-04-21

    Chrysanthemum, a leading ornamental species, does not tolerate salinity stress, although some of its related species do. The current level of understanding regarding the mechanisms underlying salinity tolerance in this botanical group is still limited. A comparison of the physiological responses to salinity stress was made between Chrysanthemum morifolium 'Jinba' and its more tolerant relatives Crossostephium chinense, Artemisia japonica and Chrysanthemum crassum. The stress induced a higher accumulation of Na(+) and more reduction of K(+) in C. morifolium than in C. chinense, C. crassum and A. japonica, which also showed higher K(+)/Na(+) ratio. Homologs of an Na(+)/H(+) antiporter (SOS1) were isolated from each species. The gene carried by the tolerant plants were more strongly induced by salt stress than those carried by the non-tolerant ones. When expressed heterologously, they also conferred a greater degree of tolerance to a yeast mutant lacking Na(+)-pumping ATPase and plasma membrane Na(+)/H(+) antiporter activity. The data suggested that the products of AjSOS1, CrcSOS1 and CcSOS1 functioned more effectively as Na (+) excluders than those of CmSOS1. Over expression of four SOS1s improves the salinity tolerance of transgenic plants and the overexpressing plants of SOS1s from salt tolerant plants were more tolerant than that from salt sensitive plants. In addition, the importance of certain AjSOS1 residues for effective ion transport activity and salinity tolerance was established by site-directed mutagenesis and heterologous expression in yeast. AjSOS1, CrcSOS1 and CcSOS1 have potential as transgenes for enhancing salinity tolerance. Some of the mutations identified here may offer opportunities to better understand the mechanistic basis of salinity tolerance in the chrysanthemum complex.

  3. The efficacy of Na-butyrate encapsulated in palm fat on performance of broilers infected with necrotic enteritis with gene expression analysis

    Directory of Open Access Journals (Sweden)

    M. G. Eshak

    2016-05-01

    Full Text Available Aim: To study the efficacy of Na-butyrate encapsulated in palm fat on performance of broiler chickens experimentally infected with necrotic enteritis (NE with the determination of its protective effect against the changes in the gene expression profiles and deoxyribonucleic acid (DNA fragmentation. Materials and Methods: A total of 800 one-day-old male Arbor Acres Plus broiler chickens were randomly allocated into four groups for 5 weeks. Na-butyrate was supplemented at dosages of 1 kg/ton for starter diet, 0.5 kg/ton for grower diet, and 0.25 kg/ton for finisher diet (presence or absence. Birds of groups 1 and 2 were inoculated by crop gavages with 4×108 CFU/ml/bird of Clostridium perfringens in phosphate buffered saline for 4 successive days, from 14 to 17 days of age to produce NE. Results: Addition of Na-butyrate, encapsulated in palm fat, to ration of experimentally infected broilers with NE resulted in increased final body weight, at 35 days of age, reduced total feed consumption, improved feed conversion ratio, reduced cumulative mortality, and increased production number. There were increased intestinal diameter, intestinal length, and significantly increased the weight of bursa of Fabricius(BF with higher hemagglutination inhibition titers against Newcastle disease (ND vaccination versus untreated infected and untreated negative control birds. The results showed increased expression levels of alpha-toxin and glyceraldehyde-3-phosphate dehydrogenase in the bursa tissues of broilers infected with C. perfringens. However, the expression levels of these genes in broilers treated with Na-butyrate were similar to the non-infected control group. Supplementation of broilers with Na-butyrate increased the expression level of insulin-like growth factor-1 (IGF-1 and decreased the DNA fragmentation induced by C. perfringens. Conclusion: Na-butyrate significantly improved chicken broiler body weights, increased relative weights of BF, increased

  4. Clinical Effectiveness of Peramivir in Comparison with Other Neuraminidase Inhibitors in Pediatric Influenza Patients

    Directory of Open Access Journals (Sweden)

    Toshiyuki Hikita

    2012-01-01

    Full Text Available The currently used antivirals in the treatment of influenza in Japan include amantadine, oseltamivir, zanamivir, laninamivir, and peramivir. We compared the efficacy of intravenous peramivir with that of other neuraminidase inhibitors for treating pediatric influenza. The present study included 223 influenza patients (≤18 years who presented at the Hikita Pediatric Clinic between February and April 2011. We compared fever duration after starting treatment with antiviral drugs. Because inhalation drugs are difficult to use in <5-year-old patients and because of the potential adverse effects of oseltamivir in teenagers, we created two different age groups (<10-year-old group and 5–18-year-old group to evaluate treatment results. In influenza A patients between 5 and 18 years old, the median fever duration after treatment with zanamivir was 2 days, compared with 1 day for peramivir (=0.0242. In influenza B patients between 5 and 18 years old, the median fever duration after treatment with laninamivir was 3 days, compared with 1 day for peramivir (=0.0097. We found no significant difference for any of the other combinations of drug/disease type/age groups. No adverse effects were observed with the antiviral drugs used. The results suggest that peramivir is very useful in pediatric influenza patients.

  5. The accuracy and timeliness of neuraminidase inhibitor dispensing data for predicting laboratory-confirmed influenza.

    Science.gov (United States)

    Papenburg, J; Charland, K M; DE Serres, G; Buckeridge, D L

    2016-06-01

    Neuraminidase inhibitor (NI) dispensing has emerged as a possible automated data source for influenza surveillance. We aimed to evaluate its timeliness, correlation, and predictive accuracy in relation to influenza activity in Quebec, Canada, 2010-2013. Our secondary objective was to use the same metrics to compare NI dispensing to visits for influenza-like illness (ILI) in emergency departments (EDs). Provincial weekly counts of positive influenza laboratory tests were used as a reference measure for the level of influenza circulation. We applied ARIMA models to account for serial correlation. We computed cross-correlations to measure the strengths of association and lead-lag relationships between NI dispensing, ILI ED visits, and our reference indicator. Finally, using an ARIMA model, we evaluated the ability of NI dispensing and ILI ED visits to predict laboratory-confirmed influenza. NI dispensing was significantly correlated (R = 0·68) with influenza activity with no lag. The maximal correlation of ILI ED visits was not as strong (R = 0·50). Both NI dispensing and ILI ED visits were significant predictors of laboratory-confirmed influenza in a multivariable model; predictive potential was greatest when NI counts were lagged to precede laboratory surveillance by 2 weeks. We conclude that NI dispensing data provides timely and valuable information for influenza surveillance.

  6. Protective Protein/Cathepsin A Rescues N-glycosylation defects in Neuraminidase-1

    Science.gov (United States)

    Wang, Dongning; Zaitsev, Slava; Taylor, Garry; d’Azzo, Alessandra; Bonten, Erik

    2009-01-01

    Background Neuraminidase-1 (NEU1) catabolizes the hydrolysis of sialic acids from sialo-glycoconjugates. NEU1 depends on its interaction with the protective protein/cathepsin A (PPCA) for lysosomal compartmentalization and catalytic activation. Murine NEU1 contains 4 N-glycosylation sites, 3 of which are conserved in the human enzyme. The expression of NEU1 gives rise to differentially glycosylated proteins. Methods We generated single-point mutations in mouse NEU1 at each of the 4 N-glycosylation sites. Mutant enzymes were expressed in NEU1-deficient cells in the presence and absence of PPCA. Results All 4 N-glycosylation variants were targeted to the lysosomal/endosomal compartment. All N-glycans, with the exception of the most C-terminal glycan, were important for maintaining stability or catalytic activity. The loss of catalytic activity caused by the deletion of the second N-glycan was rescued by increasing PPCA expression. Similar results were obtained with a human NEU1 N-glycosylation mutant identified in a sialidosis patient. Conclusions The N-terminal N-glycan of NEU1 is indispensable for its function, whereas the C-terminal N-glycan appears to be non-essential. The omission of the second N-glycan can be compensated for by upregulating the expression of PPCA. General Significance These findings could be relevant for the design of target therapies for patients carrying specific NEU1 mutations. PMID:19714866

  7. Aggregation of Streptococcus sanguis by a neuraminidase-sensitive component of serum and crevicular fluid.

    Science.gov (United States)

    Morris, E J; McBride, B C

    1983-01-01

    A number of strains of Streptococcus sanguis were found to aggregate in nonimmune serum and in crevicular fluid. All strains which aggregated in serum also aggregated in saliva, but some strains which aggregated in saliva did not aggregate in serum. Aggregation was destroyed by treatment of serum or crevicular fluid with neuraminidase and was inhibited by gangliosides. Treatment of serum with proteases reduced aggregating activity. Adsorption of serum to hydroxyapatite did not reduce the aggregating activity. The aggregating factor was partially purified by gel filtration and polyacrylamide gel electrophoresis and was found to be an acidic glycoprotein with a molecular weight of greater than 200,000, comprised of subunits with molecular weights of approximately 100,000. It did not appear to be an immunoglobulin and could not be identified with any other serum component tested. The possible role of the aggregating factor in providing nonimmune protection against colonization of S. sanguis in the gingival crevice and blood is discussed. Images PMID:6358038

  8. New Insights into Molecular Organization of Human Neuraminidase-1: Transmembrane Topology and Dimerization Ability

    Science.gov (United States)

    Maurice, Pascal; Baud, Stéphanie; Bocharova, Olga V.; Bocharov, Eduard V.; Kuznetsov, Andrey S.; Kawecki, Charlotte; Bocquet, Olivier; Romier, Beatrice; Gorisse, Laetitia; Ghirardi, Maxime; Duca, Laurent; Blaise, Sébastien; Martiny, Laurent; Dauchez, Manuel; Efremov, Roman G.; Debelle, Laurent

    2016-12-01

    Neuraminidase 1 (NEU1) is a lysosomal sialidase catalyzing the removal of terminal sialic acids from sialyloconjugates. A plasma membrane-bound NEU1 modulating a plethora of receptors by desialylation, has been consistently documented from the last ten years. Despite a growing interest of the scientific community to NEU1, its membrane organization is not understood and current structural and biochemical data cannot account for such membrane localization. By combining molecular biology and biochemical analyses with structural biophysics and computational approaches, we identified here two regions in human NEU1 - segments 139-159 (TM1) and 316-333 (TM2) - as potential transmembrane (TM) domains. In membrane mimicking environments, the corresponding peptides form stable α-helices and TM2 is suited for self-association. This was confirmed with full-size NEU1 by co-immunoprecipitations from membrane preparations and split-ubiquitin yeast two hybrids. The TM2 region was shown to be critical for dimerization since introduction of point mutations within TM2 leads to disruption of NEU1 dimerization and decrease of sialidase activity in membrane. In conclusion, these results bring new insights in the molecular organization of membrane-bound NEU1 and demonstrate, for the first time, the presence of two potential TM domains that may anchor NEU1 in the membrane, control its dimerization and sialidase activity.

  9. Estimating the Fitness Advantage Conferred by Permissive Neuraminidase Mutations in Recent Oseltamivir-Resistant A(H1N1)pdm09 Influenza Viruses

    Science.gov (United States)

    Butler, Jeff; Hooper, Kathryn A.; Petrie, Stephen; Lee, Raphael; Maurer-Stroh, Sebastian; Reh, Lucia; Guarnaccia, Teagan; Baas, Chantal; Xue, Lumin; Vitesnik, Sophie; Leang, Sook-Kwan; McVernon, Jodie; Kelso, Anne; Barr, Ian G.; McCaw, James M.; Bloom, Jesse D.; Hurt, Aeron C.

    2014-01-01

    Oseltamivir is relied upon worldwide as the drug of choice for the treatment of human influenza infection. Surveillance for oseltamivir resistance is routinely performed to ensure the ongoing efficacy of oseltamivir against circulating viruses. Since the emergence of the pandemic 2009 A(H1N1) influenza virus (A(H1N1)pdm09), the proportion of A(H1N1)pdm09 viruses that are oseltamivir resistant (OR) has generally been low. However, a cluster of OR A(H1N1)pdm09 viruses, encoding the neuraminidase (NA) H275Y oseltamivir resistance mutation, was detected in Australia in 2011 amongst community patients that had not been treated with oseltamivir. Here we combine a competitive mixtures ferret model of influenza infection with a mathematical model to assess the fitness, both within and between hosts, of recent OR A(H1N1)pdm09 viruses. In conjunction with data from in vitro analyses of NA expression and activity we demonstrate that contemporary A(H1N1)pdm09 viruses are now more capable of acquiring H275Y without compromising their fitness, than earlier A(H1N1)pdm09 viruses circulating in 2009. Furthermore, using reverse engineered viruses we demonstrate that a pair of permissive secondary NA mutations, V241I and N369K, confers robust fitness on recent H275Y A(H1N1)pdm09 viruses, which correlated with enhanced surface expression and enzymatic activity of the A(H1N1)pdm09 NA protein. These permissive mutations first emerged in 2010 and are now present in almost all circulating A(H1N1)pdm09 viruses. Our findings suggest that recent A(H1N1)pdm09 viruses are now more permissive to the acquisition of H275Y than earlier A(H1N1)pdm09 viruses, increasing the risk that OR A(H1N1)pdm09 will emerge and spread worldwide. PMID:24699865

  10. Cross-reactive neuraminidase antibodies afford partial protection against H5N1 in mice and are present in unexposed humans.

    Directory of Open Access Journals (Sweden)

    Matthew R Sandbulte

    2007-02-01

    Full Text Available BACKGROUND: A pandemic H5N1 influenza outbreak would be facilitated by an absence of immunity to the avian-derived virus in the human population. Although this condition is likely in regard to hemagglutinin-mediated immunity, the neuraminidase (NA of H5N1 viruses (avN1 and of endemic human H1N1 viruses (huN1 are classified in the same serotype. We hypothesized that an immune response to huN1 could mediate cross-protection against H5N1 influenza virus infection. METHODS AND FINDINGS: Mice were immunized against the NA of a contemporary human H1N1 strain by DNA vaccination. They were challenged with recombinant A/Puerto Rico/8/34 (PR8 viruses bearing huN1 (PR8-huN1 or avN1 (PR8-avN1 or with H5N1 virus A/Vietnam/1203/04. Additional naïve mice were injected with sera from vaccinated mice prior to H5N1 challenge. Also, serum specimens from humans were analyzed for reactivity with avN1. Immunization elicited a serum IgG response to huN1 and robust protection against the homologous challenge virus. Immunized mice were partially protected from lethal challenge with H5N1 virus or recombinant PR8-avN1. Sera transferred from immunized mice to naïve animals conferred similar protection against H5N1 mortality. Analysis of human sera showed that antibodies able to inhibit the sialidase activity of avN1 exist in some individuals. CONCLUSIONS: These data reveal that humoral immunity elicited by huN1 can partially protect against H5N1 infection in a mammalian host. Our results suggest that a portion of the human population could have some degree of resistance to H5N1 influenza, with the possibility that this could be induced or enhanced through immunization with seasonal influenza vaccines.

  11. Estimating the fitness advantage conferred by permissive neuraminidase mutations in recent oseltamivir-resistant A(H1N1pdm09 influenza viruses.

    Directory of Open Access Journals (Sweden)

    Jeff Butler

    2014-04-01

    Full Text Available Oseltamivir is relied upon worldwide as the drug of choice for the treatment of human influenza infection. Surveillance for oseltamivir resistance is routinely performed to ensure the ongoing efficacy of oseltamivir against circulating viruses. Since the emergence of the pandemic 2009 A(H1N1 influenza virus (A(H1N1pdm09, the proportion of A(H1N1pdm09 viruses that are oseltamivir resistant (OR has generally been low. However, a cluster of OR A(H1N1pdm09 viruses, encoding the neuraminidase (NA H275Y oseltamivir resistance mutation, was detected in Australia in 2011 amongst community patients that had not been treated with oseltamivir. Here we combine a competitive mixtures ferret model of influenza infection with a mathematical model to assess the fitness, both within and between hosts, of recent OR A(H1N1pdm09 viruses. In conjunction with data from in vitro analyses of NA expression and activity we demonstrate that contemporary A(H1N1pdm09 viruses are now more capable of acquiring H275Y without compromising their fitness, than earlier A(H1N1pdm09 viruses circulating in 2009. Furthermore, using reverse engineered viruses we demonstrate that a pair of permissive secondary NA mutations, V241I and N369K, confers robust fitness on recent H275Y A(H1N1pdm09 viruses, which correlated with enhanced surface expression and enzymatic activity of the A(H1N1pdm09 NA protein. These permissive mutations first emerged in 2010 and are now present in almost all circulating A(H1N1pdm09 viruses. Our findings suggest that recent A(H1N1pdm09 viruses are now more permissive to the acquisition of H275Y than earlier A(H1N1pdm09 viruses, increasing the risk that OR A(H1N1pdm09 will emerge and spread worldwide.

  12. Produção de apitoxina por abelhas Apis mellifera L. e seu efeito na expressão de genes relacionados ao estresse

    OpenAIRE

    Modanesi, Melina Stoian [UNESP

    2012-01-01

    Os objetivos do presente trabalho foram verificar o efeito do período (manhã e tarde) e tempo de colheita (30 e 60 minutos) na produção de veneno por abelhas Apis mellifera africanizadas e a influência desse manejo na expressão de genes relacionados ao estresse. Foram utilizadas cinco colméias de abelhas Apis mellifera L. As colheitas de veneno e abelhas ocorreram três vezes por semana (manhã às 09h00 e tarde às 14h00), em dias intercalados, de acordo com os seguintes tratamentos: T1: manhã/3...

  13. Computational design of drug candidates for influenza A virus subtype H1N1 by inhibiting the viral neuraminidase-1 enzyme

    Directory of Open Access Journals (Sweden)

    Tambunan Usman Sumo Friend

    2014-06-01

    Full Text Available It is critical to seek potential alternative treatments for H1N1 infections by inhibiting neuraminidase-1 enzyme. One of the viable options for inhibiting the activity of neuraminidase- 1 is peptide drug design. In order to increase peptide stability, cyclization is necessary to prevent its digestion by protease enzyme. Cyclization of peptide ligands by formation of disulfide bridges is preferable for designing inhibitors of neuraminidase-1 because of their high activity and specificity. Here we designed ligands by using molecular docking, drug scan and dynamics computational methods. Based on our docking results, short polypeptides of cystein-arginine-methionine-tyrosine- -proline-cysteine (CRMYPC and cysteine-arginine-aspargine- phenylalanine-proline-cysteine (CRNFPC have good residual interactions with the target and the binding energy ΔGbinding of -31.7402 and -31.0144 kcal mol-1, respectively. These values are much lower than those of the standards, and it means that both ligands are more accessible to ligand-receptor binding. Based on drug scan results, both of these ligands are neither mutagenic nor carcinogenic. They also show good oral bioavailability. Moreover, both ligands show relatively stable molecular dynamics progression of RMSD vs. time plot. However, based on our metods, the CRMYPC ligand has sufficient hydrogen bonding interactions with residues of the active side of neuraminidase-1 and can be therefore proposed as a potential inhibitor of neuraminidase-1

  14. Cloning and functional characterization of a vacuolar Na+/H+ antiporter gene from mungbean (VrNHX1 and its ectopic expression enhanced salt tolerance in Arabidopsis thaliana.

    Directory of Open Access Journals (Sweden)

    Sagarika Mishra

    Full Text Available Plant vacuolar NHX exchangers play a significant role in adaption to salt stress by compartmentalizing excess cytosolic Na+ into vacuoles and maintaining cellular homeostasis and ionic equilibrium. We cloned an orthologue of the vacuolar Na+/H+ antiporter gene, VrNHX1 from mungbean (Vigna radiata, an important Asiatic grain legume. The VrNHX1 (Genbank Accession number JN656211.1 contains 2095 nucleotides with an open reading frame of 1629 nucleotides encoding a predicted protein of 542 amino acids with a deduced molecular mass of 59.6 kDa. The consensus amiloride binding motif (84LFFIYLLPPI93 was observed in the third putative transmembrane domain of VrNHX1. Bioinformatic and phylogenetic analysis clearly suggested that VrNHX1 had high similarity to those of orthologs belonging to Class-I clade of plant NHX exchangers in leguminous crops. VrNHX1 could be strongly induced by salt stress in mungbean as the expression in roots significantly increased in presence of 200 mM NaCl with concomitant accumulation of total [Na+]. Induction of VrNHX1 was also observed under cold and dehydration stress, indicating a possible cross talk between various abiotic stresses. Heterologous expression in salt sensitive yeast mutant AXT3 complemented for the loss of yeast vacuolar NHX1 under NaCl, KCl and LiCl stress indicating that VrNHX1 was the orthologue of ScNHX1. Further, AXT3 cells expressing VrNHX1 survived under low pH environment and displayed vacuolar alkalinization analyzed using pH sensitive fluorescent dye BCECF-AM. The constitutive and stress inducible expression of VrNHX1 resulted in enhanced salt tolerance in transgenic Arabidopsis thaliana lines. Our work suggested that VrNHX1 was a salt tolerance determinant in mungbean.

  15. Cloning and functional characterization of a vacuolar Na+/H+ antiporter gene from mungbean (VrNHX1) and its ectopic expression enhanced salt tolerance in Arabidopsis thaliana.

    Science.gov (United States)

    Mishra, Sagarika; Alavilli, Hemasundar; Lee, Byeong-ha; Panda, Sanjib Kumar; Sahoo, Lingaraj

    2014-01-01

    Plant vacuolar NHX exchangers play a significant role in adaption to salt stress by compartmentalizing excess cytosolic Na+ into vacuoles and maintaining cellular homeostasis and ionic equilibrium. We cloned an orthologue of the vacuolar Na+/H+ antiporter gene, VrNHX1 from mungbean (Vigna radiata), an important Asiatic grain legume. The VrNHX1 (Genbank Accession number JN656211.1) contains 2095 nucleotides with an open reading frame of 1629 nucleotides encoding a predicted protein of 542 amino acids with a deduced molecular mass of 59.6 kDa. The consensus amiloride binding motif (84LFFIYLLPPI93) was observed in the third putative transmembrane domain of VrNHX1. Bioinformatic and phylogenetic analysis clearly suggested that VrNHX1 had high similarity to those of orthologs belonging to Class-I clade of plant NHX exchangers in leguminous crops. VrNHX1 could be strongly induced by salt stress in mungbean as the expression in roots significantly increased in presence of 200 mM NaCl with concomitant accumulation of total [Na+]. Induction of VrNHX1 was also observed under cold and dehydration stress, indicating a possible cross talk between various abiotic stresses. Heterologous expression in salt sensitive yeast mutant AXT3 complemented for the loss of yeast vacuolar NHX1 under NaCl, KCl and LiCl stress indicating that VrNHX1 was the orthologue of ScNHX1. Further, AXT3 cells expressing VrNHX1 survived under low pH environment and displayed vacuolar alkalinization analyzed using pH sensitive fluorescent dye BCECF-AM. The constitutive and stress inducible expression of VrNHX1 resulted in enhanced salt tolerance in transgenic Arabidopsis thaliana lines. Our work suggested that VrNHX1 was a salt tolerance determinant in mungbean.

  16. POLIMORFISMO T102C DO GENE DO RECEPTOR DE SEROTONINA (5-HT2A) NA SUSCETIBILIDADE À FIBROMIALGIA: META-ANÁLISE.

    OpenAIRE

    Peres, Paula Aparecida Borges

    2012-01-01

    Apesar de sua etiopatologia obscura, a fibromialgia é definida por dores musculoesqueléticas difusas e não articulares e sensibilidade a palpação exacerbada em determinados pontos musculares, afetando principalmente mulheres, numa proporção de 6 a 10 mulheres para cada homem. Vários estudos têm demonstrado o papel relevante dos polimorfismos de genes serotoninérgicos, dopaminérgicos e catecolaminérgicos na etiologia da fibromialgia. Desta forma, fatores genéticos podem ser d...

  17. Neuraminidase inhibitor resistance in influenza: assessing the danger of its generation and spread.

    Directory of Open Access Journals (Sweden)

    Andreas Handel

    2007-12-01

    Full Text Available Neuraminidase Inhibitors (NI are currently the most effective drugs against influenza. Recent cases of NI resistance are a cause for concern. To assess the danger of NI resistance, a number of studies have reported the fraction of treated patients from which resistant strains could be isolated. Unfortunately, those results strongly depend on the details of the experimental protocol. Additionally, knowing the fraction of patients harboring resistance is not too useful by itself. Instead, we want to know how likely it is that an infected patient can generate a resistant infection in a secondary host, and how likely it is that the resistant strain subsequently spreads. While estimates for these parameters can often be obtained from epidemiological data, such data is lacking for NI resistance in influenza. Here, we use an approach that does not rely on epidemiological data. Instead, we combine data from influenza infections of human volunteers with a mathematical framework that allows estimation of the parameters that govern the initial generation and subsequent spread of resistance. We show how these parameters are influenced by changes in drug efficacy, timing of treatment, fitness of the resistant strain, and details of virus and immune system dynamics. Our study provides estimates for parameters that can be directly used in mathematical and computational models to study how NI usage might lead to the emergence and spread of resistance in the population. We find that the initial generation of resistant cases is most likely lower than the fraction of resistant cases reported. However, we also show that the results depend strongly on the details of the within-host dynamics of influenza infections, and most importantly, the role the immune system plays. Better knowledge of the quantitative dynamics of the immune response during influenza infections will be crucial to further improve the results.

  18. Effects of vaccination and the new neuraminidase inhibitor, laninamivir, on influenza infection.

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    Takuro Mizuno

    Full Text Available BACKGROUND: Evidence of the effectiveness of influenza vaccination in children and elderly adults is limited, although this population has the highest risk for influenza infection. MATERIALS AND METHODS: We enrolled 4443 participants, aged 3-97 years, who had influenza-kit-positive results during seasons 2007-12, including 2135 with influenza A, 534 with A/H1N1, and 1643 with influenza B. Eligible subjects completed a questionnaire to identify past influenza infection and vaccination history. For the diagnosis of current influenza infection, subjects were examined, and pharyngeal swabs were collected and tested using the Capilia flu rapid diagnosis kit to confirm influenza infection. An interim analysis was performed using clinician-based surveillance data for the entire four seasons of influenza infection in Japan. RESULTS: In 3035 adults aged 14-64 years, administration of the influenza vaccine significantly reduced the frequency of infection (P65 years significantly. Laninamivir, oseltamivir phosphate, zanamivir hydrate, and amantadine hydrochloride were administered to 1381, 2432, 1044, and 100 patients, respectively. They were effective in >97% of patients, with no significant differences being found. Adverse effects were few. However, the recurrence rate of influenza infection after treatment was significantly reduced in patients who received laninamivir compared with that in those who received oseltamivir and zanamivir (P<0.01. The effectiveness of laninamivirdid not decrease. CONCLUSIONS: The vaccines administered had limited efficacy in reducing the frequency of influenza infection in young adults. Laninamivir significantly reduced the recurrence of influenza infection when compared with other neuraminidase inhibitors.

  19. Impact of AtNHX1, a vacuolar Na+/H+ antiporter, upon gene expression during short- and long-term salt stress in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Blumwald Eduardo

    2007-04-01

    Full Text Available Abstract Background AtNHX1, the most abundant vacuolar Na+/H+ antiporter in Arabidopsis thaliana, mediates the transport of Na+ and K+ into the vacuole, influencing plant development and contributing to salt tolerance. In this report, microarray expression profiles of wild type plants, a T-DNA insertion knockout mutant of AtNHX1 (nhx1, and a 'rescued' line (NHX1::nhx1 were exposed to both short (12 h and 48 h and long (one and two weeks durations of a non-lethal salt stress to identify key gene transcripts associated with the salt response that are influenced by AtNHX1. Results 147 transcripts showed both salt responsiveness and a significant influence of AtNHX1. Fifty-seven of these genes showed an influence of the antiporter across all salt treatments, while the remaining genes were influenced as a result of a particular duration of salt stress. Most (69% of the genes were up-regulated in the absence of AtNHX1, with the exception of transcripts encoding proteins involved with metabolic and energy processes that were mostly down-regulated. Conclusion While part of the AtNHX1-influenced transcripts were unclassified, other transcripts with known or putative roles showed the importance of AtNHX1 to key cellular processes that were not necessarily limited to the salt stress response; namely calcium signaling, sulfur metabolism, cell structure and cell growth, as well as vesicular trafficking and protein processing. Only a small number of other salt-responsive membrane transporter transcripts appeared significantly influenced by AtNHX1.

  20. Preterm human milk contains a large pool of latent TGF-β, which can be activated by exogenous neuraminidase

    Science.gov (United States)

    Namachivayam, Kopperuncholan; Blanco, Cynthia L.; Frost, Brandy L.; Reeves, Aaron A.; Jagadeeswaran, Ramasamy; MohanKumar, Krishnan; Safarulla, Azif; Mandal, Partha; Garzon, Steven A.; Raj, J. Usha

    2013-01-01

    Human milk contains substantial amounts of transforming growth factor (TGF)-β, particularly the isoform TGF-β2. We previously showed in preclinical models that enterally administered TGF-β2 can protect against necrotizing enterocolitis (NEC), an inflammatory bowel necrosis of premature infants. In this study we hypothesized that premature infants remain at higher risk of NEC than full-term infants, even when they receive their own mother's milk, because preterm human milk contains less bioactive TGF-β than full-term milk. Our objective was to compare TGF-β bioactivity in preterm vs. full-term milk and identify factors that activate milk-borne TGF-β. Mothers who delivered between 23 0/7 and 31 6/7 wk or at ≥37 wk of gestation provided milk samples at serial time points. TGF-β bioactivity and NF-κB signaling were measured using specific reporter cells and in murine intestinal tissue explants. TGF-β1, TGF-β2, TGF-β3, and various TGF-β activators were measured by real-time PCR, enzyme immunoassays, or established enzymatic activity assays. Preterm human milk showed minimal TGF-β bioactivity in the native state but contained a large pool of latent TGF-β. TGF-β2 was the predominant isoform of TGF-β in preterm milk. Using a combination of several in vitro and ex vivo models, we show that neuraminidase is a key regulator of TGF-β bioactivity in human milk. Finally, we show that addition of bacterial neuraminidase to preterm human milk increased TGF-β bioactivity. Preterm milk contains large quantities of TGF-β, but most of it is in an inactive state. Addition of neuraminidase can increase TGF-β bioactivity in preterm milk and enhance its anti-inflammatory effects. PMID:23558011

  1. Methanolic soluble fractions of lingzhi or reishi medicinal mushroom, Ganoderma lucidum (higher Basidiomycetes) extract inhibit neuraminidase activity in Newcastle disease virus (LaSota).

    Science.gov (United States)

    Shamaki, Bala U; Sandabe, Umar K; Ogbe, Adamu O; Abdulrahman, Fanna I; El-Yuguda, Abdul-Dahiru

    2014-01-01

    The antineuraminidase activity of different organic soluble fractions of Ganoderma lucidum extract was investigated using inhibition of hemagglutination and elution of chicken erythrocytes by Newcastle disease virus (NDV). Fractions of methanol, ethylacetate, and normal butanol (n-butanol) of the G. lucidum were tested against neuraminidase producing NDV as antigen. Different dilutions of the organic soluble fractions inhibited elution of 1% red blood cells by neuraminidase of NDV While the methanolic and n-butanol extracts inhibited neuraminidase activity even at a dilution of 1:16 and that of ethylacetate fraction inhibited even at 1:32 respectively. This finding indicates that G. lucidum has some antineuraminidase activity against NDV and may be exploited in the management of NDV infection.

  2. ANÁLISIS COMPUTACIONAL DEL EFECTO DE POLIMORFISMOS DE GENES DEL SISTEMA m-CALPAÍNA/CALPASTATINA SOBRE LA CALIDAD DE LA CARNE BOVINA

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    J. D. Leal-Gutiérrez

    2015-01-01

    Full Text Available Los genes del sistema de enzimas μ-Calpaína/Calpastatina han sido ampliamente evaluados en estudios de asociación respecto de parámetros de calidad cárnica como la terneza; previamente se han identificado varios polimorfismos asociados con la variación fenotípica en poblaciones no relacionadas de bovinos. Usando herramientas computacionales se logró postular la asociación de cuatro polimorfismos encontrados en μ-Calpaína y 11 en Calpastatina que producen una alteración de los parámetros físico-químicos, tanto del ARNm (estabilidad y polimorfismo conformacional, como de la proteína (punto isoeléctrico, potencial electroestático y superficie molecular. Es importante poder establecer el soporte biológico de polimorfismos genéticos asociados con parámetros fenotípicos que mejoren la productividad animal, lo que hace que la aproximación in silico se convierta en una herramienta útil para tal fin.

  3. Controlled clinical trial of adjuvant immunotherapy with BCG and neuraminidase-treated autologous tumour cells in large bowel cancer.

    Science.gov (United States)

    Gray, B N; Walker, C; Andrewartha, L; Freeman, S; Bennett, R C

    1989-01-01

    A controlled, randomised clinical trial of immunotherapy was performed in 301 patients with stage B or C colorectal cancer. The immunotherapy treatment consisted of 18 vaccinations over a 2 year period following surgery with a combination of BCG given by scarification plus subcutaneous injection of Vibrio cholera neuraminidase (VCN)-modified autologous tumour cells. Five year follow-up has now been completed in all patients. The immunotherapy did not alter either the disease-free interval or the overall survival of patients in comparison with a control group of patients not receiving immunotherapy.

  4. Isolation and characterization of BetaM protein encoded by ATP1B4 - a unique member of the Na,K-ATPase {beta}-subunit gene family

    Energy Technology Data Exchange (ETDEWEB)

    Pestov, Nikolay B. [Department of Physiology and Pharmacology, University of Toledo College of Medicine, 3000 Arlington Ave., Toledo, OH 43614 (United States); Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997 (Russian Federation); Zhao, Hao [Department of Physiology and Pharmacology, University of Toledo College of Medicine, 3000 Arlington Ave., Toledo, OH 43614 (United States); Basrur, Venkatesha [Department of Pathology, University of Michigan Medical School, Ann Arbor, MI 48109 (United States); Modyanov, Nikolai N., E-mail: nikolai.modyanov@utoledo.edu [Department of Physiology and Pharmacology, University of Toledo College of Medicine, 3000 Arlington Ave., Toledo, OH 43614 (United States)

    2011-09-09

    Highlights: {yields} Structural properties of BetaM and Na,K-ATPase {beta}-subunits are sharply different. {yields} BetaM protein is concentrated in nuclear membrane of skeletal myocytes. {yields} BetaM does not associate with a Na,K-ATPase {alpha}-subunit in skeletal muscle. {yields} Polypeptide chain of the native BetaM is highly sensitive to endogenous proteases. {yields} BetaM in neonatal muscle is a product of alternative splice mRNA variant B. -- Abstract: ATP1B4 genes represent a rare instance of the orthologous gene co-option that radically changed functions of encoded BetaM proteins during vertebrate evolution. In lower vertebrates, this protein is a {beta}-subunit of Na,K-ATPase located in the cell membrane. In placental mammals, BetaM completely lost its ancestral role and through acquisition of two extended Glu-rich clusters into the N-terminal domain gained entirely new properties as a muscle-specific protein of the inner nuclear membrane possessing the ability to regulate gene expression. Strict temporal regulation of BetaM expression, which is the highest in late fetal and early postnatal myocytes, indicates that it plays an essential role in perinatal development. Here we report the first structural characterization of the native eutherian BetaM protein. It should be noted that, in contrast to structurally related Na,K-ATPase {beta}-subunits, the polypeptide chain of BetaM is highly sensitive to endogenous proteases that greatly complicated its isolation. Nevertheless, using a complex of protease inhibitors, a sample of authentic BetaM was isolated from pig neonatal skeletal muscle by a combination of ion-exchange and lectin-affinity chromatography followed by SDS-PAGE. Results of the analysis of the BetaM tryptic digest using MALDI-TOF and ESI-MS/MS mass spectrometry have demonstrated that native BetaM in neonatal skeletal muscle is a product of alternative splice mRNA variant B and comprised of 351 amino acid residues. Isolated BetaM protein was

  5. Association study between mannose-binding lectin haplotypes and X gene mutation of hepatitis B virus from treatment naïve patients

    Science.gov (United States)

    Su, Chenghao; Lin, Yong; Mao, Qianguo; Wu, Daitze; Zhu, Lina; Najera, Isabel; Garcia-Alcalde, Fernando; Niu, Jianjun

    2016-01-01

    Mannose binding lectin (MBL) plays important role in the innate immunity of human. Mutations in the MBL2 gene can significantly change the serum level of MBL, and consequently alter the susceptibility and progression of infectious disease. However, the association between the MBL2 profile and the HBV mutation and quasispecies complexity has not yet been reported. Our approach includes the study of the MBL2 gene genotype as well as ultra-deep sequencing of the HBV viruses obtained from the plasma of 50 treatment naïve patients with chronic HBV infection. We found that the liver function was better among patients within the high MBL2 group with respect to those within the medium/low MBL2 group. Likewise, the number of mutations in the HBV X gene as well as the viral quasispecies complexity were significantly higher in medium/low MBL2 production group. Nucleotide substitution rates were also higher within the medium/low MBL2 production group in all positions described to have an influence in liver cancer development, except for A1499G. In this work we show that the MBL2 profile may have an impact on the HBV X gene mutations as well as on viral quasispecies complexity. PMID:27824315

  6. Synergistic Antiviral Activity of S-033188/S-033447, a Novel Inhibitor of Influenza Virus Cap-Dependent Endonuclease, in Combination with Neuraminidase Inhibitors In Vitro

    Science.gov (United States)

    Kitano, Mitsutaka; Yamamoto, Atsuko; Noshi, Takeshi; Kawai, Makoto; Yoshida, Ryu; Sato, Akihiko; Shishido, Takao; Naito, Akira

    2017-01-01

    Abstract Background S-033447, an active form of orally available prodrug S-033188, is a novel small molecule inhibitor of cap-dependent endonuclease that is essential for influenza virus transcription and replication. In this study, we evaluated the inhibitory effect of S-033188 in combination with neuraminidase inhibitors on the replication of influenza A/H1N1 virus in cultured cells. Methods The inhibitory effects of S-033447 in combination with NA inhibitors on the cytopathic effect of A/PR/8/34 strain in Madin–Darby canine kidney cells cultured for 2 days were tested and EC50 were determined. The combination index (CI), which were obtained when S-033188 and NA inhibitor were added at the closest ratio of each EC50 value, were used for the evaluation of these combinational effects (Table 1). CI values were calculated by the Chou and Talalay method, in which combinational effect were determined according to the criteria as follows: synergistic if CI ≤ 0.8, additive if 0.8 < CI < 1.2, and antagonistic if CI ≥ 1.2. CI = (DA/A + B)/DA + (DB/A + B)/DB + (DA/A + B × DB/A + B)/(DA × DB) DA: the EC50 of S-033447 DB: the EC50 of NA inhibitor DA/A + B: the concentration of S-033447 giving 50% inhibition in combination with NA inhibitor at the closest ratio of each EC50 value DB/A + B: the concentration of NA inhibitor giving 50% inhibition in combination with S-033447 at the closest ratio of each EC50 value Results All CI values were lower than 0.8, under the condition that both S-033447 and NA inhibitor (oseltamivir acid, zanamivir hydrate, laninamivir, or peramivir trihydrate) were added at the closest ratio of each EC50 value (Table 1). Conclusion S-033447 in combination with oseltamivir acid, zanamivir hydrate, laninamivir, or peramivir trihydrate synergistically inhibited the replication of influenza A/H1N1 virus in MDCK cells. Table 1. Combination effect of S-033447 and NA inhibitor in MDCK cells infected with A/PR/8/34 strain Substance A

  7. Inhibition of neuraminidase inhibitor-resistant influenza virus by DAS181, a novel sialidase fusion protein.

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    Gallen B Triana-Baltzer

    2009-11-01

    Full Text Available Antiviral drug resistance for influenza therapies remains a concern due to the high prevalence of H1N1 2009 seasonal influenza isolates which display H274Y associated oseltamivir-resistance. Furthermore, the emergence of novel H1N1 raises the potential that additional reassortments can occur, resulting in drug resistant virus. Thus, additional antiviral approaches are urgently needed. DAS181 (Fludase, a sialidase fusion protein, has been shown to have inhibitory activity against a large number of seasonal influenza strains and a highly pathogenic avian influenza (HPAI strain (H5N1. Here, we examine the in vitro activity of DAS181 against a panel of 2009 oseltamivir-resistant seasonal H1N1 clinical isolates. The activity of DAS181 against nine 2009, two 2007, and two 2004 clinical isolates of seasonal IFV H1N1 was examined using plaque number reduction assay on MDCK cells. DAS181 strongly inhibited all tested isolates. EC50 values remained constant against isolates from 2004, 2007, and 2009, suggesting that there was no change in DAS181 sensitivity over time. As expected, all 2007 and 2009 isolates were resistant to oseltamivir, consistent with the identification of the H274Y mutation in the NA gene of all these isolates. Interestingly, several of the 2007 and 2009 isolates also exhibited reduced sensitivity to zanamivir, and accompanying HA mutations near the sialic acid binding site were observed. DAS181 inhibits IFV that is resistant to NAIs. Thus, DAS181 may offer an alternative therapeutic option for seasonal or pandemic IFVs that become resistant to currently available antiviral drugs.

  8. Anti-Tumor Effects of an Oncolytic Adenovirus Expressing Hemagglutinin-Neuraminidase of Newcastle Disease Virus in Vitro and in Vivo

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    Dongyun He

    2014-02-01

    Full Text Available Oncolytic virotherapy has been an attractive drug platform for targeted therapy of cancer over the past few years. Viral vectors can be used to target and lyse cancer cells, but achieving good efficacy and specificity with this treatment approach is a major challenge. Here, we assessed the ability of a novel dual-specific anti-tumor oncolytic adenovirus, expressing the hemagglutinin-neuraminidase (HN gene from the Newcastle disease virus under the human telomerase reverse transcriptase (hTERT promoter (Ad-hTERTp-E1a-HN, to inhibit esophageal cancer EC-109 cells in culture and to reduce tumor burden in xenografted BALB/c nude mice. In vitro, infection with Ad-hTERT-E1a-HN could inhibit the growth of EC-109 cells significantly and also protect normal human liver cell line L02 from growth suppression in 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT assays. Ad-hTERT-E1a-HN also effectively and selectively decreased the sialic acid level on EC-109 cells, but not on L02 cells. Furthermore, Ad-hTERT-E1a-HN was shown to induce the apoptosis pathway via acridine orange and ethidium bromide staining (AO/EB staining, increase reactive oxygen species (ROS, reduce mitochondrial membrane potential and release cytochrome c. In vivo, xenografted BALB/c nude mice were treated via intratumoral or intravenous injections of Ad-hTERT-E1a-HN. Although both treatments showed an obvious suppression in tumor volume, only Ad-hTERT-E1a-HN delivered via intratumoral injection elicited a complete response to treatment. These results reinforced previous findings and highlighted the potential therapeutic application of Ad-hTERT-E1a-HN for treatment of esophageal cancer in clinical trials.

  9. Differential actions of proteinases and neuraminidase on mammalian erythrocyte surface and its impact on erythrocyte agglutination by concanavalin A.

    Science.gov (United States)

    Sharma, Savita; Gokhale, Sadashiv M

    2012-12-01

    Action of proteinases viz. trypsin and chymotrypsin, and neuraminidase on intact erythrocyte membrane proteins and glycophorins (sialoglycoproteins) exposed to cell surface and its impact on lectin (concanavalin A)-mediated agglutination were studied in Homo sapiens (human), Capra aegagrus hircus (goat) and Bubalus bubalis (buffalo). Membrane proteins and glycophorins analysis by SDS-PAGE as visualized by coomassie brilliant blue and periodic acid-schiff stains, respectively, and agglutination behaviour revealed marked differences: 1) there were prominent dissimilarities in the number and molecular weights of glycophorins in human, goat and buffalo erythrocyte membranes; 2) proteinase action(s) on human and buffalo erythrocyte surface membrane proteins and glycophorins showed similarity but was found different in goat; 3) significant differences in erythrocyte agglutinability with concanavalin A can be attributed to differences in membrane composition and alterations in the surface proteins after enzyme treatment; 4) a direct correlation was found between degradation of glycophorins and concanavalin A agglutinability; 5) action of neuraminidase specifically indicated the negative role of cell surface sialic acids in determining concanavalin A agglutinability of goat and buffalo erythrocytes, similar to human. Present studies clearly indicate that there are some basic differences in human, goat and buffalo erythrocyte membrane proteins, especially with respect to glycophorins, which determine the concanavalin A-mediated agglutination in enzyme treated erythrocytes.

  10. Potent bacterial neuraminidase inhibitors, anthraquinone glucosides from Polygonum cuspidatum and their inhibitory mechanism.

    Science.gov (United States)

    Uddin, Zia; Song, Yeong Hun; Curtis-Long, Marcus J; Kim, Jeong Yoon; Yuk, Heung Joo; Park, Ki Hun

    2016-12-04

    P. cuspidatum is a popular Chinese medicinal herb, having a long history of usage in traditional Chinese medicine for the treatment of several inflammatory diseases in the form of powders and decoctions. Similarly there are many reports that P. cuspidatum has antibacterial and anti-inflammatory effects, both of which are properties associated with compounds having activity against bacterial neuraminidase (BNA). We investigated whether P. cuspidatum's metabolites exhibited BNA inhibition. Consistent with our hypothesis, we found several inhibitors from the methanol extract of this plant, and then fully characterized their inhibitory mechanisms. Activity guided separation of methanol extract led to isolation of individual constituents, and subsequently their structures were elucidated by spectroscopic analysis. Detailed kinetic behaviors of BNA inhibitors were explored by showing the changes of Km and Vmax, the ratios of KI/KIS and Kik/Kiv, and fluorescence quenching effect. This study attempted to isolate the responsible metabolites and elucidate the BNA inhibitory mechanism. The principal BNA inhibitory compounds (2-6) were identified as emodin (2), physcion-8-O-β-D-glucopyranoside (3), emodin-8-O-β-D-glucopyranoside (4), emodin-1-O-β-D-glucopyranoside (5), and 2-methoxy-6-acetyl-7-methyljuglone (6). Unexpectedly, anthraquinone glucosides (3-5) were much more potent than their corresponding aglycones (1 and 2). For example, emodin (2) had an IC50=5.4μM, whereas its glucosides (4 and 5) had IC50=0.85μM and 0.43μM respectively. A similar trend was observed with physcion (1, IC50>200μM) and its glucoside (3, IC50=6.2μM). The anthraquinone (2) was mixed type I inhibitor, whereas its glucosides (4 and 5) were noncompetitive. In addition, the fluorescence quenching study showed that the affinity constants (KSV) of inhibitors increased in proportion to their inhibitory potencies. Furthermore, we quantified the major and minor metabolites through UPLC

  11. Salinity tolerance, Na+ exclusion and allele mining of HKT1;5 in Oryza sativa and O. glaberrima: many sources, many genes, one mechanism?

    Science.gov (United States)

    2013-01-01

    Background Cultivated rice species (Oryza sativa L. and O. glaberrima Steud.) are generally considered among the crop species most sensitive to salt stress. A handful of lines are known to be tolerant, and a small number of these have been used extensively as donors in breeding programs. However, these donors use many of the same genes and physiological mechanisms to confer tolerance. Little information is available on the diversity of mechanisms used by these species to cope with salt stress, and there is a strong need to identify varieties displaying additional physiological and/or genetic mechanisms to confer higher tolerance. Results Here we present data on 103 accessions from O. sativa and 12 accessions from O. glaberrima, many of which are identified as salt tolerant for the first time, showing moderate to high tolerance of high salinity. The correlation of salinity-induced senescence (as judged by the Standard Evaluation System for Rice, or SES, score) with whole-plant and leaf blade Na+ concentrations was high across nearly all accessions, and was almost identical in both O. sativa and O. glaberrima. The association of leaf Na+ concentrations with cultivar-groups was very weak, but association with the OsHKT1;5 allele was generally strong. Seven major and three minor alleles of OsHKT1;5 were identified, and their comparisons with the leaf Na+ concentration showed that the Aromatic allele conferred the highest exclusion and the Japonica allele the least. A number of exceptions to this association with the Oryza HKT1;5 allele were identified; these probably indicate the existence of additional highly effective exclusion mechanisms. In addition, two landraces were identified, one from Thailand and the other from Senegal, that show high tissue tolerance. Conclusions Significant variation in salinity tolerance exists within both cultivated Oryza species, and this is the first report of significant tolerance in O. glaberrima. The majority of accessions display a

  12. Putrescine production by Lactococcus lactis subsp. cremoris CECT 8666 is reduced by NaCl via a decrease in bacterial growth and the repression of the genes involved in putrescine production.

    Science.gov (United States)

    Del Rio, Beatriz; Redruello, Begoña; Ladero, Victor; Fernandez, Maria; Martin, Maria Cruz; Alvarez, Miguel A

    2016-09-02

    The reduction of NaCl in food is a public health priority; high NaCl intakes have been associated with serious health problems. However, it is reported that reducing the NaCl content of cheeses may lead to an increase in the content of biogenic amines (BAs). The present work examines the effect of NaCl on the accumulation of putrescine (one of the BAs often detected at high concentration in cheese) in experimental Cabrales-like cheeses containing Lactococcus lactis subsp. cremoris CECT 8666, a dairy strain that catabolises agmatine to putrescine via the agmatine deiminase (AGDI) pathway. The genes responsible for this pathway are grouped in the AGDI cluster. This comprises a regulatory gene (aguR) (transcribed independently), followed by the catabolic genes that together form an operon (aguBDAC). Reducing the NaCl concentration of the cheese led to increased putrescine accumulation. In contrast, increasing the NaCl concentration of both pH-uncontrolled and pH-controlled (pH 6) cultures of L. lactis subsp. cremoris CECT 8666 significantly inhibited its growth and the production of putrescine. Such production appeared to be inhibited via a reduction in the transcription of the aguBDAC operon; no effect on the transcription of aguR was recorded. The present results suggest that low-sodium cheeses are at risk of accumulating higher concentrations of putrescine. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. H1N1 2009 Pandemic Influenza Virus: Resistance of the I223R Neuraminidase Mutant Explained by Kinetic and Structural Analysis

    NARCIS (Netherlands)

    E. van der Vries (Erhard); P.J. Collins (Patrick ); S.G. Vachieri (Sebastien); X. Xiong (Xiaoli); J. Liu (Jinhua); P.A. Walker (Philip); L.F. Haire (Lesley ); A.J. Hay (Alan); M. Schutten (Martin); A.D.M.E. Osterhaus (Albert); S.R. Martin (Steve ); C.A. Boucher (Charles); J.J. Skehel (John ); S.J. Gamblin (Steve )

    2012-01-01

    textabstractTwo classes of antiviral drugs, neuraminidase inhibitors and adamantanes, are approved for prophylaxis and therapy against influenza virus infections. A major concern is that antiviral resistant viruses emerge and spread in the human population. The 2009 pandemic H1N1 virus is already

  14. Two single mutations in the fusion protein of Newcastle disease virus confer hemagglutinin-neuraminidase independent fusion promotion and attenuate the pathogenicity in chickens

    Science.gov (United States)

    The fusion (F) protein of Newcastle disease virus (NDV) plays an important role in viral infection and pathogenicity through mediating membrane fusion between the virion and host cells in the presence of the hemagglutinin-neuraminidase (HN). Previously, we obtained a velogenic NDV genotype VII muta...

  15. Relationship between Interleukin-6 gene polymorphism and hippocampal volume in antipsychotic-naïve schizophrenia: evidence for differential susceptibility?

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    Sunil Vasu Kalmady

    Full Text Available Various lines of evidence including epidemiological, genetic and foetal pathogenetic models suggest a compelling role for Interleukin-6 (IL-6 in the pathogenesis of schizophrenia. IL-6 mediated inflammatory response triggered by maternal infection or stress induces disruption of prenatal hippocampal development which might contribute towards psychopathology during adulthood. There is a substantial lack of knowledge on how genetic predisposition to elevated IL-6 expression effects hippocampal structure in schizophrenia patients. In this first-time study, we evaluated the relationship between functional polymorphism rs1800795 of IL-6 and hippocampal gray matter volume in antipsychotic-naïve schizophrenia patients in comparison with healthy controls.We examined antipsychotic-naïve schizophrenia patients [N = 28] in comparison with healthy controls [N = 37] group matched on age, sex and handedness. Using 3 Tesla - MRI, bilateral hippocampi were manually segmented by blinded raters with good inter-rater reliability using a valid method. Additionally, Voxel-based Morphometry (VBM analysis was performed using hippocampal mask. The IL-6 level was measured in blood plasma using ELISA technique. SNP rs1800795 was genotyped using PCR and DNA sequencing. Psychotic symptoms were assessed using Scale for Assessment of Positive Symptoms and Scale for Assessment of Negative Symptoms.Schizophrenia patients had significantly deficient left and right hippocampal volumes after controlling for the potential confounding effects of age, sex and total brain volume. Plasma IL-6 levels were significantly higher in patients than controls. There was a significant diagnosis by rs1800795 genotype interaction involving both right and left hippocampal volumes. Interestingly, this effect was significant only in men but not in women.Our first time observations suggest a significant relationship between IL-6 rs1800795 and reduced hippocampal volume in antipsychotic-na

  16. Proteome analysis and functional expression identify mortalin as an antiapoptotic gene induced by elevation of [Na+]i/[K+]i ratio in cultured vascular smooth muscle cells.

    Science.gov (United States)

    Taurin, Sebastien; Seyrantepe, Volkan; Orlov, Sergei N; Tremblay, Tammy-Lynn; Thibault, Pierre; Bennett, Martin R; Hamet, Pavel; Pshezhetsky, Alexey V

    2002-11-15

    Apoptosis of vascular smooth muscle cells (VSMCs) plays an important role in remodeling of vessel walls, one of the major determinants of long-term blood pressure elevation and an independent risk factor for cardiovascular morbidity and mortality. Recently, we have found that apoptosis in cultured VSMCs can be inhibited by inversion of the intracellular [Na+]/[K+] ratio after the sustained blockage of the Na+,K+-ATPase by ouabain. To understand the mechanism of ouabain action, we analyzed subsets of hydrophilic and hydrophobic VSMC proteins from control and ouabain-treated cells by 2-dimensional electrophoresis. Ouabain treatment led to overexpression of numerous soluble and hydrophobic cellular proteins. Among proteins that showed the highest level of ouabain-induced expression, we identified mortalin (also known as GRP75 or PBP-74), a member of the heat shock protein 70 (HSP70) superfamily and a marker for cellular mortal and immortal phenotypes. Northern and Western blotting and immunocytochemistry all have confirmed that treatment of VSMCs with ouabain results in potent induction of mortalin expression. Transient transfection of cells with mortalin cDNA led to at least a 6-hour delay in the development of apoptosis after serum deprivation. The expression of tumor suppressor gene, p53, in mortalin-transfected cells was delayed to the same extent, and the expressed protein showed abnormal perinuclear distribution, suggesting that p53 is retained and inactivated by mortalin. Our studies therefore define a new [Na+]i/[K+]i-responsive signaling pathway that may play an important role in the regulation of programmed cell death in VSMCs.

  17. HIV reverse transcriptase gene mutations in anti-retroviral treatment naïve rural people living with HIV/AIDS

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    K Mohanakrishnan

    2015-01-01

    Full Text Available This study is designed to find out the mutational variations of reverse transcriptase (RT gene of HIV, after the traditional drug usage among anti-retroviral therapy naïve rural people living with HIV/AIDS. HIV Reactive patients, who were exposed for indigenous medicines such as Siddha, Ayurveda etc., for a minimum period of 6 months were taken for this study. Among 40 patients, two samples (5.55% demonstrated high-level mutational resistance variations for nucleoside RT inhibitor (NRTI and non-NRTI. The predominant polymorphisms detected were K122E (91.7%, V60I (91.7%, V35T (89%, Q207E (89%, D177E (89%, T200A (86.1%, S48T (83.33%, K173A (80.6%.

  18. Effects of SCN9A gene modification on Na+ channel and the expression of nerve growth factor in a rat model of diarrhea‑predominant irritable bowel syndrome.

    Science.gov (United States)

    Cai, Yong-Yan; Li, Chen; Yan, Zhi-Xin; Ma, Na; Li, Fang-Fang

    2018-01-01

    The aim of the present study was to identify whether the sodium voltage-gated channel alpha subunit 9 (SCN9A) gene modification is a potential treatment for diarrhea‑predominant irritable bowel syndrome (D‑IBS), via regulating the Na+ channel and the expression of nerve growth factor (NGF). The recombinant adenovirus vector of the SCN9A gene was established, and rat colon cells were isolated for SCN9A gene modification. All subjects were divided into four groups: i) The SCN9A‑modified (D‑IBS rat model implanted with SCN9A‑modified colon cells), ii) negative control (NC; D‑IBS rat model implanted with colon cells without SCN9A gene modification), iii) blank (D‑IBS rat model without any treatment) and iv) normal (normal rats without any treatment). Western blotting and reverse transcription‑quantitative polymerase chain reaction were used to detect the protein and mRNA expression levels of SCN9A, NGF and voltage gated sodium channels (Nav)1.8 and Nav1.9 in rat colon tissues. Compared with the normal group, the rats in the SCN9A, NC and blank groups had significantly elevated mRNA and protein expression levels of NGF, SCN9A, Nav1.8 and Nav1.9. The rats in the SCN9A group demonstrated significantly increased mRNA and protein expression levels of NGF, SCN9A, Nav1.8 and Nav1.9 compared with the NC group and the blank group (all PNav1.9 channels, in addition to NGF which may provide a novel therapeutic basis for treating of D‑IBS.

  19. Neuraminidase inhibitors for preventing and treating influenza in healthy adults and children

    Directory of Open Access Journals (Sweden)

    Tom Jefferson

    Full Text Available BACKGROUND:Neuraminidase inhibitors (NIs are stockpiled and recommended by public health agencies for treating and preventing seasonal and pandemic influenza. They are used clinically worldwide.OBJECTIVE:To describe the potential benefits and harms of NIs for influenza in all age groups by reviewing all clinical study reports of published and unpublished randomised, placebo-controlled trials and regulatory comments.METHODSSearch methods: We searched trial registries, electronic databases (to 22 July 2013 and regulatory archives, and corresponded with manufacturers to identify all trials. We also requested clinical study reports. We focused on the primary data sources of manufacturers but we checked that there were no published randomised controlled trials (RCTs from non-manufacturer sources by running electronic searches in the following databases: the Cochrane Central Register of Controlled Trials (CENTRAL, MEDLINE, MEDLINE (Ovid, EMBASE, Embase.com, PubMed (not MEDLINE, the Database of Reviews of Effects, the NHS Economic Evaluation Database and the Health Economic Evaluations Database.Selection criteria: Randomised, placebo-controlled trials on adults and children with confirmed or suspected exposure to naturally occurring influenza.Data collection and analysis: We extracted clinical study reports and assessed risk of bias using purpose-built instruments. We analysed the effects of zanamivir and oseltamivir on time to first alleviation of symptoms, influenza outcomes, complications, hospitalisations and adverse events in the intention-to-treat (ITT population. All trials were sponsored by the manufacturers.MAIN RESULTS: We obtained 107 clinical study reports from the European Medicines Agency (EMA, GlaxoSmithKline and Roche. We accessed comments by the US Food and Drug Administration (FDA, EMA and Japanese regulator. We included 53 trials in Stage 1 (a judgement of appropriate study design and 46 in Stage 2 (formal analysis, including 20

  20. Neuraminidase inhibitors for influenza: a systematic review and meta-analysis of regulatory and mortality data.

    Science.gov (United States)

    Heneghan, Carl J; Onakpoya, Igho; Jones, Mark A; Doshi, Peter; Del Mar, Chris B; Hama, Rokuro; Thompson, Matthew J; Spencer, Elizabeth A; Mahtani, Kamal R; Nunan, David; Howick, Jeremy; Jefferson, Tom

    2016-01-01

    BACKGROUND Neuraminidase inhibitors (NIs) are stockpiled and recommended by public health agencies for treating and preventing seasonal and pandemic influenza. They are used clinically worldwide. OBJECTIVES To (1) describe the potential benefits and harms of NIs for influenza in all age groups by reviewing all clinical study reports (CSRs) of published and unpublished randomised, placebo-controlled trials and regulatory comments; and (2) determine the effect of oseltamivir (Tamiflu(®), Roche) treatment on mortality in patients with 2009A/H1N1 influenza. METHODS We searched trial registries, electronic databases and corresponded with regulators and sponsors to identify randomised trials of NIs. We requested full CSRs and accessed regulators' comments. We included only those trials for which we had CSRs. To examine the effects of oseltamivir on 2009A/H1N1 influenza mortality, we requested individual patient data (IPD) from corresponding authors of all included observational studies. RESULTS Effect of oseltamivir and zanamivir (Relenza®, GlaxoSmithKline) in the prevention and treatment of influenza: Oseltamivir reduced the time to first alleviation of symptoms in adults by 16.8 hours [95% confidence interval (CI) 8.4 to 25.1 hours]. Zanamivir reduced the time to first alleviation of symptoms in adults by 0.60 days (95% CI 0.39 to 0.81 days). Oseltamivir reduced unverified pneumonia in adult treatment [risk difference (RD) 1.00%, 95% CI 0.22% to 1.49%]; similar findings were observed with zanamivir prophylaxis in adults (RD 0.32%, 95% CI 0.09% to 0.41%). Oseltamivir treatment of adults increased the risk of nausea (RD 3.66%, 95% CI 0.90% to 7.39%) and vomiting (RD 4.56%, 95% CI 2.39% to 7.58%). In the treatment of children, oseltamivir induced vomiting (RD 5.34%, 95% CI 1.75% to 10.29%). Both oseltamivir and zanamivir prophylaxis reduced the risk of symptomatic influenza in individuals (oseltamivir RD 3.05%, 95% CI 1.83% to 3.88%; zanamivir RD 1.98%, 95% CI 0.98% to

  1. Differential responses of CO2 assimilation, carbohydrate allocation and gene expression to NaCl stress in perennial ryegrass with different salt tolerance.

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    Tao Hu

    Full Text Available Little is known about the effects of NaCl stress on perennial ryegrass (Lolium perenne L. photosynthesis and carbohydrate flux. The objective of this study was to understand the carbohydrate metabolism and identify the gene expression affected by salinity stress. Seventy-four days old seedlings of two perennial ryegrass accessions (salt-sensitive 'PI 538976' and salt-tolerant 'Overdrive' were subjected to three levels of salinity stress for 5 days. Turf quality in all tissues (leaves, stems and roots of both grass accessions negatively and significantly correlated with GFS (Glu+Fru+Suc content, except for 'Overdrive' stems. Relative growth rate (RGR in leaves negatively and significantly correlated with GFS content in 'Overdrive' (P0.05 for turf quality. A greater up-regulation in the expression of SPS, SS, SI, 6-SFT gene was observed in 'Overdrive' than 'PI 538976'. A higher level of SPS and SS expression in leaves was found in 'PI 538976' relative to 'Overdrive'. Accumulation of hexoses in roots, stems and leaves can induce a feedback repression to photosynthesis in salt-stressed perennial ryegrass and the salt tolerance may be changed with the carbohydrate allocation in leaves and stems.

  2. Active 1918 pandemic flu viral neuraminidase has distinct N-glycan profile and is resistant to trypsin digestion.

    Science.gov (United States)

    Wu, Zhengliang L; Ethen, Cheryl; Hickey, Gregg E; Jiang, Weiping

    2009-02-13

    The 1918 pandemic flu virus caused one of the most deadly pandemics in human history. To search for unique structural features of the neuraminidase from this virus that might have contributed to its unusual virulence, we expressed this enzyme. The purified enzyme appeared as a monomer, a dimer and a tetramer, with only the tetramer being active and therefore biologically relevant. The monomer and the dimer could not be oligomerized into the tetramer in solution, suggesting that some unique structural features were required for oligomerization and activation. These features could be related to N-glycosylation, because the tetramer displayed different N-glycans than the monomer and the dimer. Furthermore, the tetramer was found to be resistant to trypsin digestion, which may give the virus the capability to invade tissues that are normally not infected by influenza viruses and make the virus more robust for infection.

  3. Regulation of ENA1 Na+-ATPase Gene Expression by the Ppz1 Protein Phosphatase Is Mediated by the Calcineurin Pathway

    Science.gov (United States)

    Ruiz, Amparo; Yenush, Lynne; Ariño, Joaquín

    2003-01-01

    Saccharomyces cerevisiae strains lacking the Ppz1 protein phosphatase are salt tolerant and display increased expression of the ENA1 Na+-ATPase gene, a major determinant for sodium extrusion, while cells devoid of the similar Ppz2 protein do not show these phenotypes. However, a ppz1 ppz2 mutant displays higher levels of ENA1 expression than the ppz1 strain. We show here that the increased activity of the ENA1 promoter in a ppz1 ppz2 mutant maps to two regions: one region located at −751 to −667, containing a calcineurin-dependent response element (CDRE), and one downstream region (−573 to −490) whose activity responds to intracellular alkalinization. In contrast, the increased ENA1 expression in a ppz1 mutant is mediated solely by an intact calcineurin/Crz1 signaling pathway, on the basis that (i) this effect maps to a single region that contains the CDRE and (ii) it is blocked by the calcineurin inhibitor FK506, as well as by deletion of the CNB1 or CRZ1 gene. The calcineurin dependence of the increased ENA1 expression of a ppz1 mutant would suggest that Ppz1 could negatively regulate calcineurin activity. In agreement with this notion, a ppz1 strain is calcium sensitive, and this mutation does not result in a decrease in the calcium hypertolerance of a cnb1 mutant. It has been shown that ENA1 can be induced by alkalinization of the medium and that a ppz1 ppz2 strain has a higher intracellular pH. However, we present several lines of evidence that show that the gene expression profile of a ppz1 mutant does not involve an alkalinization effect. In conclusion, we have identified a novel role for calcineurin, but not alkalinization, in the control of ENA1 expression in ppz1 mutants. PMID:14555476

  4. Fluxo gênico em soja na Região Oeste do Paraná Soybean gene flow in the Western Region of Paraná

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    Ivan Schuster

    2007-04-01

    Full Text Available Este trabalho teve o objetivo de avaliar o fluxo gênico em soja, na Região Oeste do Paraná. Foram semeados cinco círculos concêntricos, com a cultivar CD 219RR, que contém o gene CP4 EPSPS. Os círculos foram espaçados em 50 cm, com círculo interno de diâmetro de 50 cm. Externamente a estes, foi semeada a cultivar CD 211 (convencional, também em cinco círculos concêntricos, espaçados em 1 m. As plantas da cultivar CD 211 foram colhidas e trilhadas individualmente, e as sementes semeadas novamente no campo. Após a emergência, foram obtidas 151.772 plântulas, as quais, com 15 dias, foram pulverizadas com 900 g ha-1 de i.a. de glifosato. Após uma semana, plantas sobreviventes foram submetidas à análise de PCR, para verificar a presença do gene CP4 EPSPS. A taxa de fecundação cruzada foi de 0,61, 0,29, 0,23, 0,22 e 0,23% respectivamente a 1, 2, 3, 4 e 5 m de distância das plantas geneticamente modificadas.The objective of this work was to evaluate soybean gene flow in the Western Region of Paraná. Five concentric circles were sowed with the CD 219RR cultivar, which contains the CP4 EPSPS gene. The circles were spaced in 50 cm and the central circle had 50 cm in diameter. Externally to the CD 219RR circles, five concentric circles were sowed with CD 211 cultivar, a no genetically modified soybean, spaced of 1 m. The CD 211 plants were harvested and threshed separately and the seeds were sowed again. After the emergency, 151,772 seedlings were obtained, which with 15 days were sprayed with 900 g ha-1 a.i. of glyphosate. After one week, the surviving plants were analysed by PCR to verify the CP4 EPSPS gene presence. The cross-pollinating rate was 0.61, 0.29, 0.23, 0.22 and 0.23% in 1, 2, 3, 4 and 5 m distance of the genetically modified plants, respectively.

  5. Gene

    Data.gov (United States)

    U.S. Department of Health & Human Services — Gene integrates information from a wide range of species. A record may include nomenclature, Reference Sequences (RefSeqs), maps, pathways, variations, phenotypes,...

  6. mrpA, a gene with roles in resistance to Na+ and adaptation to alkaline pH in the cyanobacterium Anabaena sp. PCC7120.

    Science.gov (United States)

    Blanco-Rivero, A; Leganés, F; Fernández-Valiente, E; Calle, P; Fernández-Piñas, F

    2005-05-01

    Transposon mutagenesis of Anabaena sp. PCC7120 led to the isolation of a mutant strain, PHB11, which grew poorly at pH values above 10. The mutant strain exhibited pronounced Na+ sensitivity; this sensitivity was higher under basic conditions. Mutant PHB11 also showed an inhibition of photosynthesis that was much more pronounced at alkaline pH. Reconstruction of the transposon mutation of PHB11 in the wild-type strain reproduced the phenotype of the original mutant. The wild-type version of the mutated gene was cloned and the mutation complemented. In mutant strain PHB11, the transposon had inserted within an ORF that is part of a seven-ORF operon with significant sequence similarity to a family of bacterial operons that are believed to code for a novel multiprotein cation/proton antiporter primarily involved in resistance to salt stress and adaptation to alkaline pH. The Anabaena operon was denoted mrp (multiple resistance and pH adaptation) following the nomenclature of the Bacillus subtilis operon; the ORF mutated in PHB11 corresponded to mrpA. Computer analysis suggested that all seven predicted Anabaena Mrp proteins were highly hydrophobic with several transmembrane domains; in fact, the predicted protein sequences encoded by mrpA, mrpB and mrpC showed significant similarity to hydrophobic subunits of the proton pumping NADH : ubiquinone oxidoreductase. In vivo expression studies indicated that mrpA is induced with increasing external Na+ concentrations and alkaline pH; mrpA is also upregulated under inorganic carbon (Ci) limitation. The biological significance of a putative cyanobacterial Mrp complex is discussed.

  7. Genes Я us” – Of juis nie? Oor determinisme en voluntarisme by die mens met verwysing na homoseksualiteit

    Directory of Open Access Journals (Sweden)

    Johan Buitendag

    2004-11-01

    Full Text Available “Genes Я us” – or not? About human determinism and voluntarism, with reference to homosexualityThis article has as its departure point the conviction of some that human genome mapping predisposes human beings genetically and as a consequence, the homosexual person becomes a mere victim of circumstances. Biological determinism and social construc-tionism are not mutually exclusive and although a person is orientated within a web of boundary matters, the depiction of a human being as imago Dei still prevails. A person has the freedom to choose and the responsibility to do so. One’s understanding of reality provides a frame of reference from which a definition of morality is derived. The suggestion of Nancey Murphey to understand reality as a “nonreductive physicalism” is followed. Reductionism in any form is subsequently avoided. A holistic view of humankind in terms of which religious experience is seen as more than some brain functions and people are embedded in a “sacred canopy”, is therefore advocated.

  8. GABAergic gene expression in postmortem hippocampus from alcoholics and cocaine addicts; corresponding findings in alcohol-naïve P and NP rats.

    Science.gov (United States)

    Enoch, Mary-Anne; Zhou, Zhifeng; Kimura, Mitsuru; Mash, Deborah C; Yuan, Qiaoping; Goldman, David

    2012-01-01

    By performing identical studies in humans and rats, we attempted to distinguish vulnerability factors for addiction from neurobiological effects of chronic drug exposure. We focused on the GABAergic system within the hippocampus, a brain region that is a constituent of the memory/conditioning neuronal circuitry of addiction that is considered to be important in drug reinforcement behaviors in animals and craving and relapse in humans. Using RNA-Seq we quantified mRNA transcripts in postmortem total hippocampus from alcoholics, cocaine addicts and controls and also from alcohol-naïve, alcohol preferring (P) and non-preferring (NP) rats selectively bred for extremes of alcohol-seeking behavior that also show a general addictive tendency. A pathway-targeted analysis of 25 GABAergic genes encoding proteins implicated in GABA synthesis, metabolism, synaptic transmission and re-uptake was undertaken. Directionally consistent and biologically plausible overlapping and specific changes were detected: 14/25 of the human genes and 12/25 of the rat genes showed nominally significant differences in gene expression (global p values: 9×10⁻¹⁴, 7×10⁻¹¹ respectively). Principal FDR-corrected findings were that GABBR1 was down-regulated in alcoholics, cocaine addicts and P rats with congruent findings in NSF, implicated in GABAB signaling efficacy, potentially resulting in increased synaptic GABA. GABRG2, encoding the gamma2 subunit required for postsynaptic clustering of GABAA receptors together with GPHN, encoding the associated scaffolding protein gephryin, were both down-regulated in alcoholics and cocaine addicts but were both up-regulated in P rats. There were also expression changes specific to cocaine addicts (GAD1, GAD2), alcoholics (GABRA2) and P rats (ABAT, GABRG3). Our study confirms the involvement of the GABAergic system in alcoholism but also reveals a hippocampal GABA input in cocaine addiction. Congruent findings in human addicts and P rats provide clues

  9. Computational model for analyzing the evolutionary patterns of the neuraminidase gene of influenza A/H1N1.

    Science.gov (United States)

    Ahn, Insung; Son, Hyeon Seok

    2012-02-01

    In this study, we performed computer simulations to evaluate the changes of selection potentials of codons in influenza A/H1N1 from 1999 to 2009. We artificially generated the sequences by using the transition matrices of positively selected codons over time, and their similarities against the database of influenzavirus A genus were determined by BLAST search. This is the first approach to predict the evolutionary direction of influenza A virus (H1N1) by simulating the codon substitutions over time. We observed that the BLAST results showed the high similarities with pandemic influenza A/H1N1 in 2009, suggesting that the classical human-origin influenza A/H1N1 isolated before 2009 might contain some selection potentials of swine-origin viruses. Computer simulations using the time series codon substitution patterns resulted dramatic changes of BLAST results in influenza A/H1N1, providing a possibility of developing a method for predicting the viral evolution in silico. Copyright © 2011 Elsevier Ltd. All rights reserved.

  10. Identification of five driver gene mutations in patients with treatment-naïve lung adenocarcinoma in Taiwan.

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    Kuo-Hsuan Hsu

    Full Text Available It is important to select appropriate targeted therapies for subgroups of patients with lung adenocarcinoma who have specific gene alterations.This prospective study was a multicenter project conducted in Taiwan for assessment of lung adenocarcinoma genetic tests. Five oncogenic drivers, including EGFR, KRAS, BRAF, HER2 and EML4-ALK fusion mutations, were tested. EGFR, KRAS, BRAF and HER2 mutations were assessed by MALDI-TOF MS (Cohort 1. EML4-ALK translocation was tested by Ventana method in EGFR-wild type patients (Cohort 2.From August 2011 to November 2013, a total of 1772 patients with lung adenocarcinoma were enrolled. In Cohort 1 analysis, EGFR, KRAS, HER2 and BRAF mutations were identified in 987 (55.7%, 93 (5.2%, 36 (2.0% and 12 (0.7% patients, respectively. Most of these mutations were mutually exclusive, except for co-mutations in seven patients (3 with EGFR + KRAS, 3 with EGFR + HER2 and 1 with KRAS + BRAF. In Cohort 2 analysis, 29 of 295 EGFR-wild type patients (9.8% were positive for EML4-ALK translocation. EGFR mutations were more common in female patients and non-smokers and KRAS mutations were more common in male patients and smokers. Gender and smoking status were not correlated significantly with HER2, BRAF and EML4-ALK mutations. EML4-ALK translocation was more common in patients with younger age.This was the first study in Taiwan to explore the incidence of five oncogenic drivers in patients with lung adenocarcinoma and the results could be valuable for physicians in consideration of targeted therapy and inclusion of clinical trials.

  11. Optimization of influenza A vaccine virus by reverse genetic using chimeric HA and NA genes with an extended PR8 backbone.

    Science.gov (United States)

    Medina, Julie; Boukhebza, Houda; De Saint Jean, Amélie; Sodoyer, Régis; Legastelois, Isabelle; Moste, Catherine

    2015-08-20

    The yield of influenza antigen production may significantly vary between vaccine strains; for example the A/California/07/09 (H1N1)-X179A vaccine virus, prepared during 2009 influenza pandemic, presented a low antigen yield in eggs compared to other seasonal H1N1 reassortants. In this study a bi-chimeric virus expressing HA and NA genes with A/Puerto Rico/8/34 (H1N1) (PR8) and X179A domains was rescued by reverse genetics using a mixture of Vero/CHOK1 cell lines (Medina et al. [7]). The bi-chimeric virus obtained demonstrated to yield much larger amounts of HA than X179A in eggs as measured by single-radial-immunodiffusion (SRID), the reference method to quantify HA protein in influenza vaccine. Such kind of optimized virus using PR8 backbone derived chimeric glycoproteins could be used as improved seed viruses for vaccine production. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Regulation of cation transporter genes by the arbuscular mycorrhizal symbiosis in rice plants subjected to salinity suggests improved salt tolerance due to reduced Na(+) root-to-shoot distribution.

    Science.gov (United States)

    Porcel, Rosa; Aroca, Ricardo; Azcon, Rosario; Ruiz-Lozano, Juan Manuel

    2016-10-01

    Rice is a salt-sensitive crop whose productivity is strongly reduced by salinity around the world. Plants growing in saline soils are subjected to the toxicity of specific ions such as sodium, which damage cell organelles and disrupt metabolism. Plants have evolved biochemical and molecular mechanisms to cope with the negative effects of salinity. These include the regulation of genes with a role in the uptake, transport or compartmentation of Na(+) and/or K(+). Studies have shown that the arbuscular mycorrhizal (AM) symbiosis alleviates salt stress in several host plant species. However, despite the abundant literature showing mitigation of ionic imbalance by the AM symbiosis, the molecular mechanisms involved are barely explored. The objective of this study was to elucidate the effects of the AM symbiosis on the expression of several well-known rice transporters involved in Na(+)/K(+) homeostasis and measure Na(+) and K(+) contents and their ratios in different plant tissues. Results showed that OsNHX3, OsSOS1, OsHKT2;1 and OsHKT1;5 genes were considerably upregulated in AM plants under saline conditions as compared to non-AM plants. Results suggest that the AM symbiosis favours Na(+) extrusion from the cytoplasm, its sequestration into the vacuole, the unloading of Na(+) from the xylem and its recirculation from photosynthetic organs to roots. As a result, there is a decrease of Na(+) root-to-shoot distribution and an increase of Na(+) accumulation in rice roots which seems to enhance the plant tolerance to salinity and allows AM rice plants to maintain their growing processes under salt conditions.

  13. In vitro neuraminidase inhibitory concentration (IC50) of four neuraminidase inhibitors against clinical isolates of the influenza viruses circulating in the 2010-2011 to 2014-2015 Japanese influenza seasons.

    Science.gov (United States)

    Ikematsu, Hideyuki; Kawai, Naoki; Iwaki, Norio; Kashiwagi, Seizaburo

    2016-09-01

    To assess the extent of viral resistance to the four neuraminidase inhibitors (NAIs), we measured their 50% inhibitory concentration (IC50) for influenza virus isolates from the 2014-2015 influenza season for comparison with those circulating in the 2010-2011 to 2013-2014 influenza seasons. Viral isolation was done with specimens obtained prior to treatment, and the type and subtype of influenza was determined by RT-PCR using type- and subtype-specific primers. The IC50 was determined by a neuraminidase inhibition assay using a fluorescent substrate. IC50 was measured for 200 influenza A(H3N2) and 19 influenza B in the 2014-2015 season, and no virus with highly reduced sensitivity to the four NAIs was detected. The ratios of the geometric means of the A(H3N2) IC50s of 2014-2015 to those of the 2010-2011, 2011-2012, 2012-2013, and 2013-2014 seasons ranged from 0.72 to 1.05, 0.82 to 1.22, 0.69 to 1.00, and 0.70 to 1.03, respectively. The ratios of the geometric mean of the B IC50s to the previous four seasons ranged from 0.59 to 1.28, 0.66 to 1.34, 0.84 to 1.21, and 1.06 to 1.47, respectively. There was no trend in the change of the IC50s for A(H3N2) or B. Significant differences were found in some seasons, but the differences in the IC50s were all less than two fold. These results show change in the geometric mean IC50 by season but with no trend, which indicates that the influence of viral mutation on the effectiveness of these NAIs was minute for A(H3N2) and B over the past five seasons. Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  14. H1N1 2009 pandemic influenza virus: resistance of the I223R neuraminidase mutant explained by kinetic and structural analysis.

    Directory of Open Access Journals (Sweden)

    Erhard van der Vries

    2012-09-01

    Full Text Available Two classes of antiviral drugs, neuraminidase inhibitors and adamantanes, are approved for prophylaxis and therapy against influenza virus infections. A major concern is that antiviral resistant viruses emerge and spread in the human population. The 2009 pandemic H1N1 virus is already resistant to adamantanes. Recently, a novel neuraminidase inhibitor resistance mutation I223R was identified in the neuraminidase of this subtype. To understand the resistance mechanism of this mutation, the enzymatic properties of the I223R mutant, together with the most frequently observed resistance mutation, H275Y, and the double mutant I223R/H275Y were compared. Relative to wild type, K(M values for MUNANA increased only 2-fold for the single I223R mutant and up to 8-fold for the double mutant. Oseltamivir inhibition constants (K(I increased 48-fold in the single I223R mutant and 7500-fold in the double mutant. In both cases the change was largely accounted for by an increased dissociation rate constant for oseltamivir, but the inhibition constants for zanamivir were less increased. We have used X-ray crystallography to better understand the effect of mutation I223R on drug binding. We find that there is shrinkage of a hydrophobic pocket in the active site as a result of the I223R change. Furthermore, R223 interacts with S247 which changes the rotamer it adopts and, consequently, binding of the pentoxyl substituent of oseltamivir is not as favorable as in the wild type. However, the polar glycerol substituent present in zanamivir, which mimics the natural substrate, is accommodated in the I223R mutant structure in a similar way to wild type, thus explaining the kinetic data. Our structural data also show that, in contrast to a recently reported structure, the active site of 2009 pandemic neuraminidase can adopt an open conformation.

  15. Neuraminidase-1, a Subunit of the Cell Surface Elastin Receptor, Desialylates and Functionally Inactivates Adjacent Receptors Interacting with the Mitogenic Growth Factors PDGF-BB and IGF-2

    Science.gov (United States)

    Hinek, Aleksander; Bodnaruk, Tetyana D.; Bunda, Severa; Wang, Yanting; Liu, Kela

    2008-01-01

    We recently established that the elastin-binding protein, which is identical to the spliced variant of β-galactosidase, forms a cell surface-targeted complex with two proteins considered “classic lysosomal enzymes”: protective protein/cathepsin A and neuraminidase-1 (Neu1). We also found that cell surface-residing Neu1 can desialylate neighboring microfibrillar glycoproteins and facilitate the deposition of insoluble elastin, which contributes to the maintenance of cellular quiescence. Here we provide evidence that cell surface-residing Neu1 contributes to a novel mechanism that limits cellular proliferation by desialylating cell membrane-residing sialoglycoproteins that directly propagate mitogenic signals. We demonstrated that treatment of cultured human aortic smooth muscle cells (SMCs) with either a sialidase inhibitor or an antibody that blocks Neu1 activity induced significant up-regulation in SMC proliferation in response to fetal bovine serum. Conversely, treatment with Clostridium perfringens neuraminidase (which is highly homologous to Neu1) decreased SMC proliferation, even in cultures that did not deposit elastin. Further, we found that pretreatment of aortic SMCs with exogenous neuraminidase abolished their mitogenic responses to recombinant platelet-derived growth factor (PDGF)-BB and insulin-like growth factor (IGF)-2 and that sialidosis fibroblasts (which are exclusively deficient in Neu1) were more responsive to PDGF-BB and IGF-2 compared with normal fibroblasts. Furthermore, we provide direct evidence that neuraminidase caused the desialylation of both PDGF and IGF-1 receptors and diminished the intracellular signals induced by the mitogenic ligands PDGF-BB and IGF-2. PMID:18772331

  16. Synthesis of a cluster-forming sialylthio-D-galactose fullerene conjugate and evaluation of its interaction with influenza virus hemagglutinin and neuraminidase.

    Science.gov (United States)

    Tollas, Szilvia; Bereczki, Ilona; Borbás, Anikó; Batta, Gyula; Vanderlinden, Evelien; Naesens, Lieve; Herczegh, Pál

    2014-06-01

    In order to obtain self assembling, multivalent ligand for influenza virus hemagglutinin α-N-acetylneuraminyl-(2-6)-D-galactopyranose has been synthesized and bonded to a water soluble fullerene derivative using 1,3-dipolar cycloaddition click reaction. The aggregating amphiphilic compound did not inhibit the influenza virus hemagglutinin, but it proved to be an inhibitor of its neuraminidase with a 50% inhibitory concentration of 81 μM. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Neuraminidase inhibitors in the treatment and post exposure prevention of influenza

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    von der Schulenburg, Johann-Matthias

    2006-01-01

    Full Text Available Introduction: Influenza is a viral respiratory infection which presents itself as an acute febrile disease. It is contracted by virus-laden respiratory secretions from infected individuals. Symptoms usually last three to seven days and are accompanied by severely limited activities during this time. A definite diagnosis, however, can only be made by laboratory analysis. Every year, about 20% of children and 5% of adults develop symptomatic influenza of the serotypes A or B worldwide. Typical complications of influenza include viral or bacterial infections, as well as deterioration of an existing cardio-vascular or respiratory disease which may lead to hospitalization and death. Current policy recommends that individuals, who are at-risk of developing serious complications (patients over sixty years of age or patients with concomitant chronic diseases, as well as people in direct contact with high risk patients (i.e. nursing staff in living and care facilities, should be annually vaccinated with inactivated influenza strains. Various pharmaceutical agents for the treatment and prophylaxis of influenza have been approved. Amantadine, which inhibits the viral M2-ion channel, is only effective in influenza-serotype A. Neuraminidase inhibitors (NI represent a new class of antivirals for prophylaxis and treatment of influenza A and B. NI interrupt various central functions that are vital for the life cycle and spreading of the virus. Two drugs of this substance class, Zanamivir (RelenzaTM and Oseltamivir (Tamiflu®, are licensed for the treatment of influenza. For adults and teenagers over thirteen years of age Oseltamivir is also approved for the prophylaxis of influenza. Zanamivir is a powder which needs to be inhaled, whereas Oseltamivir is licensed as a capsule for oral administration. M2-inhibitors and NI are only effective at an early stage of the influenza infection, i.e. during the first 36 to 48 hours after symptom onset, before replication

  18. Caracterização do gene vip3A e toxicidade da proteína Vip3Aa50 à lagarta-do-cartucho e à lagarta-da-soja

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    Camila Soares Figueiredo

    2013-09-01

    Full Text Available O objetivo deste trabalho foi caracterizar o gene vip3A de Bacillus thuringiensis e verificar a toxicidade da proteína Vip3Aa50 a larvas da lagarta-do-cartucho (Spodoptera frugiperda e da lagarta-da-soja (Anticarsia gemmatalis. O gene vip3A foi amplificado por PCR, com iniciadores específicos, e gerou um fragmento de 2.370 pb. Esse fragmento foi clonado em vetor pGEM-T Easy e, em seguida, sequenciado, subclonado em vetor de expressão pET-28a (+ e inserido em células de Escherichia coli BL21 (DE3. A expressão da proteína Vip3Aa50 foi induzida por isopropil-β-D-1-tiogalactopiranosídeo (IPTG, visualizada em SDS-PAGE e detectada por "Western blot". Os ensaios de toxicidade revelaram alta atividade da proteína Vip3Aa50 contra as larvas neonatas da lagarta-da-soja e da lagarta-do-cartucho, com CL50 de 20,3 e 79,6 ng cm-2, respectivamente. O gene vip3Aa50 é um novo gene da classe vip3A.

  19. Neuraminidase inhibitors for preventing and treating influenza in healthy adults and children.

    Science.gov (United States)

    Jefferson, Tom; Jones, Mark A; Doshi, Peter; Del Mar, Chris B; Hama, Rokuro; Thompson, Matthew J; Spencer, Elizabeth A; Onakpoya, Igho; Mahtani, Kamal R; Nunan, David; Howick, Jeremy; Heneghan, Carl J

    2014-04-10

    Neuraminidase inhibitors (NIs) are stockpiled and recommended by public health agencies for treating and preventing seasonal and pandemic influenza. They are used clinically worldwide. To describe the potential benefits and harms of NIs for influenza in all age groups by reviewing all clinical study reports of published and unpublished randomised, placebo-controlled trials and regulatory comments. We searched trial registries, electronic databases (to 22 July 2013) and regulatory archives, and corresponded with manufacturers to identify all trials. We also requested clinical study reports. We focused on the primary data sources of manufacturers but we checked that there were no published randomised controlled trials (RCTs) from non-manufacturer sources by running electronic searches in the following databases: the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, MEDLINE (Ovid), EMBASE, Embase.com, PubMed (not MEDLINE), the Database of Reviews of Effects, the NHS Economic Evaluation Database and the Health Economic Evaluations Database. Randomised, placebo-controlled trials on adults and children with confirmed or suspected exposure to naturally occurring influenza. We extracted clinical study reports and assessed risk of bias using purpose-built instruments. We analysed the effects of zanamivir and oseltamivir on time to first alleviation of symptoms, influenza outcomes, complications, hospitalisations and adverse events in the intention-to-treat (ITT) population. All trials were sponsored by the manufacturers. We obtained 107 clinical study reports from the European Medicines Agency (EMA), GlaxoSmithKline and Roche. We accessed comments by the US Food and Drug Administration (FDA), EMA and Japanese regulator. We included 53 trials in Stage 1 (a judgement of appropriate study design) and 46 in Stage 2 (formal analysis), including 20 oseltamivir (9623 participants) and 26 zanamivir trials (14,628 participants). Inadequate reporting put most of the

  20. Microarray and suppression subtractive hybridization analyses of gene expression in hybrid poplar (Populus alba × Populus tremula var. glandulosa) cell suspension cultures after exposure to NaCl.

    Science.gov (United States)

    Bae, Eun-Kyung; Lee, Hyoshin; Lee, Jae-Soon; Noh, Eun-Woon; Choi, Young-Im; Lee, Byung-Hyun; Choi, Dong-Woog

    2012-09-01

    The gene expression profiles of hybrid poplar (Populus alba × Populus tremula var. glandulosa) cells in suspension culture after exposure to salinity (NaCl) induced stress were examined by constructing two suppression subtractive hybridization (SSH) libraries. cDNA from non-treated cells was used as a driver and cDNA samples from cell suspension cultures exposed to 150 mM NaCl for 2 or 10 h were used as testers. Randomly selected clones from each SSH library were sequenced and 727 high-quality expressed sequence tags (ESTs) were obtained and analyzed. Four novel ESTs were identified. Between the two libraries, 542 unique SSH clones were selected for placement on a cDNA microarray. In total, 18 differentially expressed genes were identified with 4 and 12 genes being significantly differentially expressed 2 and 10 h after the treatment, respectively. Genes related to metabolism and protein synthesis and several genes whose protein products are implicated in salt or other abiotic stress-related responses were expressed in the salt-stressed cells. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

  1. Neuraminidase Inhibition Primes Short-Term Depression and Suppresses Long-Term Potentiation of Synaptic Transmission in the Rat Hippocampus

    Directory of Open Access Journals (Sweden)

    Alina Savotchenko

    2015-01-01

    Full Text Available Neuraminidase (NEU is a key enzyme that cleaves negatively charged sialic acid residues from membrane proteins and lipids. Clinical and basic science studies have shown that an imbalance in NEU metabolism or changes in NEU activity due to various pathological conditions parallel with behavior and cognitive impairment. It has been suggested that the decreases of NEU activity could cause serious neurological consequences. However, there is a lack of direct evidences that modulation of endogenous NEU activity can impair neuronal function. Using combined rat entorhinal cortex/hippocampal slices and a specific inhibitor of NEU, 2-deoxy-2,3-dehydro-N-acetylneuraminic acid (NADNA, we examined the effect of downregulation of NEU activity on different forms of synaptic plasticity in the hippocampal CA3-to-CA1 network. We show that NEU inhibition results in a significant decrease in long-term potentiation (LTP and an increase in short-term depression. Synaptic depotentiation restores LTP in NADNA-pretreated slices to the control level. These data suggest that short-term NEU inhibition produces the LTP-like effect on neuronal network, which results in damping of further LTP induction. Our findings demonstrate that downregulation of NEU activity could have a major impact on synaptic plasticity and provide a new insight into the cellular mechanism underlying behavioral and cognitive impairment associated with abnormal metabolism of NEU.

  2. Polimorfismos do gene MASP2 e concentrações séricas da proteína MASP-2 na febre reumática e cardiopatia reumática crônica

    OpenAIRE

    Catarino, Sandra Jeremias dos Santos

    2014-01-01

    Orientador: Prof. Dr. Iara José de Mesias Reasson Orientador: Prof. Dr. Angelica B. Winter Boldt Dissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências da Saúde, Programa de Pós-Graduação em Medicina Interna. Defesa : Curitiba, 23/05/2014 Inclui referências Linha de pesquisa: Associação genética a doenças Resumo: MASP-2, a serina protease 2 associada à lectina ligante de manose ou ficolinas, é uma proteína-chave na ativação da via das lectinas do complemento...

  3. Polymorphism of alpha s1-casein gene in a dairy goat herd in the southeastern region of Brazil Polimorfismo do gene da alpha s1-caseína em um rebanho leiteiro na região sudeste do Brasil

    Directory of Open Access Journals (Sweden)

    Maria Amélia Menck Soares

    2009-06-01

    Full Text Available Three different regions of the alpha s1-casein gene (CSN1S1 were investigated to determine the frequencies of major alleles for null, low, intermediate and high milk protein expression in a herd of dairy goats raised in the southeastern region of Brazil. Genomic DNA samples were obtained from leukocytes of 145 dairy goats and regions of interest in the gene were amplified through Polymerase Chain Reaction (PCR, then evaluated in both agarose (O and E allele and polyacrylamide gels (F allele. For better characterization of the F allele, a PCR-RFLP (PCR-Restriction Fragment Length Polymorphism study was performed employing the endonuclease XmnI. The allelic frequencies in the herd of 62 Saanen goats studied were: CSN1S1E = 0.35; CSN1S1F = 0.30; CSN1S1O1 = 0.02; CSN1S1A+B+C = 0.30, other alleles = 0.03. In another group of 83 Alpine animals, the frequencies were: CSN1S1E = 0.48; CSN1S1F = 0.28; CSN1S1O1 = 0.01; CSN1S1A+B+C = 0.20, other alleles = 0.03. In the region of exon 9 and intron downstream, where mutations that characterize the F allele occur, it was verified that different intragenic haplotypes may exist, involving the deletion of the 23rd nucleotide in the ninth exon in addition to the insertion of 11bp on intron. These haplotypes may be used to make direct association with other alleles. Although rare, a higher number of combinations were found in this work by evaluating in conjunction the region of the insertion of 3bp in the referred intron, which may allow a higher number of associations. A complete characterization of these combinations will allow elaborating simplified protocols to identify animals concerning the alleles of CSN1S1 gene in goats.Foram investigadas três regiões do gene da alpha s1-caseína (CSN1S1 para determinar as frequências dos principais alelos para nula, baixa, média e alta expressão desta proteína no leite em um rebanho de cabras leiteiras na Região Sudeste do Brasil. O DNA genômico foi isolado a partir

  4. Modeling the Association of Space, Time, and Host Species with Variation of the HA, NA, and NS Genes of H5N1 Highly Pathogenic Avian Influenza Viruses Isolated from Birds in Romania in 2005–2007

    Science.gov (United States)

    Alkhamis, Mohammad; Perez, Andres; Batey, Nicole; Howard, Wendy; Baillie, Greg; Watson, Simon; Franz, Stephanie; Focosi-Snyman, Raffaella; Onita, Iuliana; Cioranu, Raluca; Turcitu, Mihai; Kellam, Paul; Brown, Ian H.; Breed, Andrew C.

    2014-01-01

    SUMMARY Molecular characterization studies of a diverse collection of avian influenza viruses (AIVs) have demonstrated that AIVs’ greatest genetic variability lies in the HA, NA, and NS genes. The objective here was to quantify the association between geographical locations, periods of time, and host species and pairwise nucleotide variation in the HA, NA, and NS genes of 70 isolates of H5N1 highly pathogenic avian influenza virus (HPAIV) collected from October 2005 to December 2007 from birds in Romania. A mixed-binomial Bayesian regression model was used to quantify the probability of nucleotide variation between isolates and its association with space, time, and host species. As expected for the three target genes, a higher probability of nucleotide differences (odds ratios [ORs] > 1) was found between viruses sampled from places at greater geographical distances from each other, viruses sampled over greater periods of time, and viruses derived from different species. The modeling approach in the present study maybe useful in further understanding the molecular epidemiology of H5N1 HPAI virus in bird populations. The methodology presented here will be useful in predicting the most likely genetic distance for any of the three gene segments of viruses that have not yet been isolated or sequenced based on space, time, and host species during the course of an epidemic. PMID:24283126

  5. Cloning, expression and sequence diversity of iss gene from avian pathogenic Escherichia coli (APEC isolated in Brazil / Clonagem, expressão e diversidade na seqüência do gene iss de Escherichia coli patogênica para aves (APEC, isolada no Brasil

    Directory of Open Access Journals (Sweden)

    Marilda Carlos Vidotto

    2010-09-01

    Full Text Available A proteína Iss (increased serum survival é uma importante característica de resistência ao sistema complemento da Escherichia coli patogênica para aves (APEC. Os objetivos deste trabalho foram clonar e verificar a diversidade da seqüência do gene iss de APEC e caracterizar a proteína Iss recombinante. O gene iss de 309 bp foi amplificado por PCR, clonado e expresso na E. coli BL21 (DE3 utilizando o vetor pET SUMO. O gene iss da APEC9 foi classificado como iss tipo 1 pela diferenciação entre 3 tipos de iss alelos. A proteína Iss foi expressa pela indução com IPTG, purificada em coluna com resina ligada ao íon níquel e utilizada na imunização de galinhas poedeiras. Anticorpos da classe IgY anti rIss reagiram com a proteina rIss, a qual apresentou massa molecular de 22 kDa, correspondendo 11kDa da Iss e 11 kDa da proteína SUMO. The Iss (Increased serum survival protein is an important characteristic of resistance to complement system of avian pathogenic Escherichia coli (APEC. The objectives of this work were to cloning and verify the sequence diversity of iss gene from APEC and characterize the recombinant Iss protein. The iss gene of 309 bp was amplified by PCR, cloned and expressed in E. coli BL21 (DE3 using the pET SUMO vector. The iss gene from APEC9 strain was classified as iss type 1 by differentiation of the three iss gene allele types. The protein was expressed by induction of IPTG and purified in resin charged with the nickel ion. Antibodies IgY anti rIss reacted with rIss showing a molecular mass of 22 kDa, corresponding 11KDa of Iss protein and 11 KDa SUMO protein.

  6. NaCl stress induces CsSAMs gene expression in Cucumis sativus by mediating the binding of CsGT-3b to the GT-1 element within the CsSAMs promoter.

    Science.gov (United States)

    Wang, Li-Wei; He, Mei-Wen; Guo, Shi-Rong; Zhong, Min; Shu, Sheng; Sun, Jin

    2017-05-01

    The CsSAMs promoter is a salt-stress-inducible promoter containing three GT-1 elements that are sufficient for the salt-stress response. The transcription factor CsGT-3b was found to bind to the GT-1 element. The S-adenosyl-L-methionine synthase (SAMs) gene is among the functional genes induced during environmental stress. However, little is known about the regulatory mechanism and upstream regulators of this salt-inducible gene in cucumber plants. Thus, it is necessary to understand the characteristics of the SAMs gene by analyzing its promoter and transcription factors. In this study, we isolated and functionally analyzed a 1743-bp flanking fragment of the CsSAMs gene from Cucumis sativus. To examine promoter activity, the full-length promoter, as well as different promoter fragments, were fused to the β-glucuronidase (GUS) reporter gene and introduced into the tobacco genome. The full-length promoter displayed maximal promoter activity, whereas the P4 promoter, containing 321 bp of upstream sequence, showed no basal promoter activity. In addition, the CsSAMs promoter exhibited stress-inducible regulation rather than tissue-specific activity in transgenic tobacco. Histochemical analysis revealed strong GUS staining in leaves, stems, and roots, especially in the veins of leaves, the vascular bundle of stems, and root tip zones following NaCl stress. A transient expression assay confirmed that the 242-bp region (-1743 to -1500) was sufficient for the NaCl-stress response. Yeast one-hybrid assays further revealed interaction between the NaCl-response protein CsGT-3b and the GT-1 (GAAAAA) element within the 242-bp region. Taken together, we revealed the presence of four salt-stress-responsive elements (GT-1 cis-elements) in the CsSAMs promoter and identified a transcription factor, CsGT-3b, that specifically binds to this sequence. These results might help us better understand the intricate regulatory network of the cucumber SAMs gene.

  7. Using Common Spatial Distributions of Atoms to Relate Functionally Divergent Influenza Virus N10 and N11 Protein Structures to Functionally Characterized Neuraminidase Structures, Toxin Cell Entry Domains, and Non-Influenza Virus Cell Entry Domains

    Science.gov (United States)

    Weininger, Arthur; Weininger, Susan

    2015-01-01

    The ability to identify the functional correlates of structural and sequence variation in proteins is a critical capability. We related structures of influenza A N10 and N11 proteins that have no established function to structures of proteins with known function by identifying spatially conserved atoms. We identified atoms with common distributed spatial occupancy in PDB structures of N10 protein, N11 protein, an influenza A neuraminidase, an influenza B neuraminidase, and a bacterial neuraminidase. By superposing these spatially conserved atoms, we aligned the structures and associated molecules. We report spatially and sequence invariant residues in the aligned structures. Spatially invariant residues in the N6 and influenza B neuraminidase active sites were found in previously unidentified spatially equivalent sites in the N10 and N11 proteins. We found the corresponding secondary and tertiary structures of the aligned proteins to be largely identical despite significant sequence divergence. We found structural precedent in known non-neuraminidase structures for residues exhibiting structural and sequence divergence in the aligned structures. In N10 protein, we identified staphylococcal enterotoxin I-like domains. In N11 protein, we identified hepatitis E E2S-like domains, SARS spike protein-like domains, and toxin components shared by alpha-bungarotoxin, staphylococcal enterotoxin I, anthrax lethal factor, clostridium botulinum neurotoxin, and clostridium tetanus toxin. The presence of active site components common to the N6, influenza B, and S. pneumoniae neuraminidases in the N10 and N11 proteins, combined with the absence of apparent neuraminidase function, suggests that the role of neuraminidases in H17N10 and H18N11 emerging influenza A viruses may have changed. The presentation of E2S-like, SARS spike protein-like, or toxin-like domains by the N10 and N11 proteins in these emerging viruses may indicate that H17N10 and H18N11 sialidase-facilitated cell

  8. Associação de SNPs nos genes para κ-caseína e β-lactoglobulina com curvas de lactação em cabras leiteiras

    Directory of Open Access Journals (Sweden)

    Samir Julián Calvo Cardona

    2015-03-01

    Full Text Available O objetivo deste trabalho foi identificar o modelo com melhor ajuste aos dados de produção de leite, gordura, proteína e sólidos totais para cabras leiteiras, bem como associar polimorfismos de nucleotídeo único (SNPs nos genes para κ-caseína (κ-CSN3 e β-lactoglobulina (β-LG aos parâmetros das curvas de produção e qualidade do leite. Foram avaliados 4.160 registros de produção de leite, gordura, proteína e sólidos totais de cabras das raças Alpina, Saanen e mestiça, no Estado de Antioquia, na Colômbia. Os modelos não lineares com melhor ajuste à estrutura dos dados foram os de Nelder, para produção de leite, e de Cappio-Borlino, para qualidade do leite. As análises de associação mostraram efeito significativo do SNP κ-CSN3 sobre o pico de produção, a produção inicial e a persistência das características qualitativas do leite. Já o SNP β-LG apresentou efeito significativo sobre os picos de produção de leite e gordura, e sobre o tempo necessário para atingir o pico de produção de proteína. A identificação e a estimação da influência dos marcadores SNP avaliados sobre as curvas de lactação e a qualidade do leite podem contribuir para a seleção de caprinos leiteiros.

  9. Salt and cadmium stress tolerance caused by overexpression of the Glycine Max Na+/H+ Antiporter (GmNHX1) gene in duckweed (Lemna turionifera 5511).

    Science.gov (United States)

    Yang, Lin; Han, Yujie; Wu, Di; Yong, Wang; Liu, Miaomiao; Wang, Sutong; Liu, Wenxin; Lu, Meiyi; Wei, Ying; Sun, Jinsheng

    2017-11-01

    Cadmium (Cd) pollution has aroused increasing attention due to its toxicity. It has been proved that Na+/H+ Antiporter (NHX1) encodes a well-documented protein in Na+/H+ trafficking, which leads to salt tolerance. This study showed that Glycine max Na+/H+ Antiporter (GmNHX1) improved short-term cadmium and salt resistance in Lemna turionifera 5511. Expression of GmNHX1 prevented root from abscission and cell membrane damage, which also can enhance antioxidant system, inhibited of reactive oxygen species (ROS) accumulation and cause a less absorption of Cd under cadmium and salt stress. The cadmium tolerance suggested that NHX1 was involved under the cadmium stress. Copyright © 2017. Published by Elsevier B.V.

  10. Single Gene Deletions of mrpA to mrpG and mrpE Point Mutations Affect Activity of the Mrp Na+/H+ Antiporter of Alkaliphilic Bacillus and Formation of Hetero-Oligomeric Mrp Complexes▿ †

    OpenAIRE

    Morino, Masato; Natsui, Shinsuke; Swartz, Talia H.; Krulwich, Terry A.; Ito, Masahiro

    2008-01-01

    Mrp antiporters catalyze secondary Na+(Li+)/H+ antiport and/or K+/H+ antiport that is physiologically important in diverse bacteria. An additional capacity for anion flux has been observed for a few systems. Mrp is unique among antiporters in that it requires all six or seven hydrophobic gene products (MrpA to MrpG) of the mrp operon for full antiporter activity, but MrpE has been reported to be dispensable. Here, the membrane complexes formed by Mrp proteins were examined using a cloned mrp ...

  11. Simple, intuitive calculations of free energy of binding for protein-ligand complexes. 2. Computational titration and pH effects in molecular models of neuraminidase-inhibitor complexes.

    Science.gov (United States)

    Fornabaio, Micaela; Cozzini, Pietro; Mozzarelli, Andrea; Abraham, Donald J; Kellogg, Glen E

    2003-10-09

    One factor that can strongly influence predicted free energy of binding is the ionization state of functional groups on the ligands and at the binding site at which calculations are performed. This analysis is seldom performed except in very detailed computational simulations. In this work, we address the issues of (i) modeling the complexity resulting from the different ionization states of ligand and protein residues involved in binding, (ii) if, and how, computational methods can evaluate the pH dependence of ligand inhibition constants, and (iii) how to score the protonation-dependent models. We developed a new and fairly rapid protocol called "computational titration" that enables parallel modeling of multiple ionization ensembles for each distinct protonation level. Models for possible protonation combinations for site/ligand ionizable groups are built, and the free energy of interaction for each of them is quantified by the HINT (Hydropathic INTeractions) software. We applied this procedure to the evaluation of the binding affinity of nine inhibitors (six derived from 2,3-didehydro-2-deoxy-N-acetylneuraminic acid, DANA) of influenza virus neuraminidase (NA), a surface glycoprotein essential for virus replication and thus a pharmaceutically relevant target for the design of anti-influenza drugs. The three-dimensional structures of the NA enzyme-inhibitor complexes indicate considerable complexity as the ligand-protein recognition site contains several ionizable moieties. Each computational titration experiment reveals a peak HINT score as a function of added protons. This maximum HINT score indicates the optimum pH (or the optimum protonation state of each inhibitor-protein binding site) for binding. The pH at which inhibition is measured and/or crystals were grown and analyzed can vary from this optimum. A protonation model is proposed for each ligand that reconciles the experimental complex structure with measured inhibition and the free energy of binding

  12. Virus-like particles displaying H5, H7, H9 hemagglutinins and N1 neuraminidase elicit protective immunity to heterologous avian influenza viruses in chickens.

    Science.gov (United States)

    Pushko, Peter; Tretyakova, Irina; Hidajat, Rachmat; Zsak, Aniko; Chrzastek, Klaudia; Tumpey, Terrence M; Kapczynski, Darrell R

    2017-01-15

    Avian influenza (AI) viruses circulating in wild birds pose a serious threat to public health. Human and veterinary vaccines against AI subtypes are needed. Here we prepared triple-subtype VLPs that co-localized H5, H7 and H9 antigens derived from H5N1, H7N3 and H9N2 viruses. VLPs also contained influenza N1 neuraminidase and retroviral gag protein. The H5/H7/H9/N1/gag VLPs were prepared using baculovirus expression. Biochemical, functional and antigenic characteristics were determined including hemagglutination and neuraminidase enzyme activities. VLPs were further evaluated in a chicken AI challenge model for safety, immunogenicity and protective efficacy against heterologous AI viruses including H5N2, H7N3 and H9N2 subtypes. All vaccinated birds survived challenges with H5N2 and H7N3 highly pathogenic AI (HPAI) viruses, while all controls died. Immune response was also detectable after challenge with low pathogenicity AI (LPAI) H9N2 virus suggesting that H5/H7/H9/N1/gag VLPs represent a promising approach for the development of broadly protective AI vaccine. Copyright © 2016. Published by Elsevier Inc.

  13. Bovine papillomavirus type 4 L1 gene transfection in a Drosophila S2 cell expression system: absence of L1 protein expression Transfecção em células de Drosophila S2 usando o gene L1 do Papilomavírus Bovino tipo 4: ausência de expressão da proteína L1

    Directory of Open Access Journals (Sweden)

    Luiz Gustavo Bentim Góes

    2008-03-01

    Full Text Available The development of a bovine papillomavirus (BPV vaccine is an outstanding challenge. BPV protein L1 gene transfection in the Drosophila melanogaster S2 cell expression system failed to produce L1 protein notwithstanding correct L1 gene insertion. Severe genetic inbalance in the host cell line, including cytogenetic alterations, may account for the lack of protein expression.O desenvolvimento de uma vacina para papilomavirus bovino (BPV consiste em grande desafio. A transfecção do gene codificante da proteína L1 de BPV em sistema de células S2 de Drosophila melanogaster não logrou sucesso, apesar da correta inserção da seqüência gênica em vetor apropriado.Graves alterações genéticas na linhagem celular S2, que incluem aberrações cromossômicas, provavelmente estão relacionadas à ausência da expressão da proteína desejada.

  14. Niveles de homocisteína y polimorfismos de los genes de la MTHFR y la CBS en pacientes colombianos con trombosis venosa superficial y profunda

    Directory of Open Access Journals (Sweden)

    Claudia Ayala

    2010-08-01

    Conclusiones. No se encontraron diferencias estadísticamente significativas en los niveles de homocisteína entre el grupo de casos y de controles. Sin embargo, la variabilidad en las concentraciones plasmáticas fue mayor en los casos.

  15. Enhanced salt tolerance conferred by the complete 2.3 kb cDNA of the rice vacuolar Na+/H+antiporter gene compared to 1.9 kb coding region with 5' UTR in transgenic lines of Rice

    Directory of Open Access Journals (Sweden)

    U.S. Mahzabin eAmin

    2016-01-01

    Full Text Available Soil salinity is one of the most challenging problems that restricts the normal growth and production of rice worldwide. It has therefore become very important to produce more saline tolerant rice varieties. This study shows constitutive over-expression of the vacuolar Na+/H+ antiporter gene (OsNHX1 from the rice landrace (Pokkali and attainment of enhanced level of salinity tolerance in transgenic rice plants. It also shows that inclusion of the complete un-translated regions of the alternatively spliced OsNHX1 gene provides a higher level of tolerance to the transgenic rice. Two separate transformation events of the OsNHX1 gene, one with 1.9 kb region containing the 5ʹ UTR with CDS and the other of 2.3 kb, including 5ʹ UTR, CDS and the 3ʹ UTR regions were performed. The transgenic plants with these two different constructs were advanced to the T3 generation and physiological and molecular screening of homozygous plants was conducted at seedling and reproductive stages under salinity (NaCl stress. Both transgenic lines were observed to be tolerant compared to WT plants at both physiological stages. However, the transgenic lines containing the CDS with both the 5ʹ and 3ʹ UTR were significantly more tolerant compared to the transgenic lines containing OsNHX1 gene without the 3′ UTR. At the seedling stage at 12 dS/m stress, the chlorophyll content was significantly higher (P 1.9 kb> and WT lines. Yield in g/plant in the best line from the 2.3 kb plants was significantly more (P<0.01 compared respectively to the best 1.9 kb line and WT plants at stress of 6 dS/m. Transformation with the complete transcripts rather than the CDS may therefore provide more durable level of tolerance.

  16. Sequence diversity in the coat protein gene of Lettuce big-vein associated virus and Mirafiori lettuce big-vein virus infecting lettuce in Brazil Variabilidade genética na porção codificadora para a proteína capsidial do Lettuce big-vein associated virus e Mirafiori lettuce big-vein virus provenientes de alface no Brasil

    Directory of Open Access Journals (Sweden)

    Márcio Martinello Sanches

    2008-06-01

    Full Text Available Lettuce big vein associated virus (LBVaV and Mirafiori lettuce big vein virus (MLBVV have been found in mixed infection in Brazil causing the lettuce big vein disease. Analysis of part of the coat protein (CP gene of Brazilian isolates of LBVaV collected from lettuce, showed at least 93% amino acid sequence identity with other LBVaV isolates. Genetic diversity among MLBVV CP sequences was higher when compared to LBVaV CP sequences, with amino acid sequence identity ranging between 91% to 100%. Brazilian isolates of MLBVV belong to subgroup A, with one RsaI restriction site on the coat protein gene. There is no indication for a possible geografical origin for the Brazilian isolates of LBVaV and MLBVV.Lettuce big vein associated virus (LBVaV e Mirafiori lettuce big vein virus (MLBVV têm sido encontrados em infecções mistas no Brasil, causando a doença conhecida como engrossamento das nervuras da alface. Análise de parte do gene da proteína capsidial (CP de isolados brasileiros de LBVaV coletados em alface, indicou que estes possuem identidade superior a 93% com isolados coletados em diferentes regiões geográficas. A diversidade genética entre a CP de isolados de MLBVV de alface foi maior comparada às sequências da CP de LBVaV, com a identidade de aminoácidos variando entre 91 a 100%. Os isolados brasileiros de MLBVV pertencem ao subgrupo A, com um único sítio de restrição RsaI no gene da proteína capsidial. Não há indicação para uma provável origem geográfica dos isolados brasileiros de MLBVV e LBVaV.

  17. AVALIAÇÃO DOS NÍVEIS DE PROTEÍNA C REATIVA CIRCULANTE E DO SNP rs1205 DO GENE CRP COM OBESIDADE, CARACTERÍSTICAS ANTROPOMÉTRICAS E MARCADORES BIOQUÍMICOS

    Directory of Open Access Journals (Sweden)

    Elisa Inês Klinger

    2015-12-01

    Full Text Available A obesidade é um grave pro¬blema de saúde pública e está relacionada a um baixo grau de inflamação crô¬nica. A Proteína C Reativa (PCR é caracterizada como proteína de fase aguda, mas em resposta à inflamação e em estados crônicos, pode assumir um papel pró-inflamatório, como no caso da obesidade. Para tanto, teve-se por objetivo avaliar a possível relação dos níveis de PCR circulante e do SNP rs1205 do gene CRP com obesidade, características antropométricas e marcadores bioquímicos em escolares de Santa Cruz do Sul – RS. Este estudo caracteriza-se por ser transversal retrospectivo, sendo realizado com 380 escolares de 07 a 17 anos de idade. A genotipagem do polimorfismo de nucleotídeo único (SNP rs1205C/T foi realizada através de PCR em tempo real. Foram coletados dados antropométricos e realizados testes bioquímicos. O tratamento estatístico foi realizado por estatística descritiva, Kruskal-Wallis, Mann-Whitney e regressão logística multivariada. Com este estudo, foi observada prevalência de 25,3% de sobrepeso e obesidade nos escolares estudados, e que portadores do alelo de risco (T apresentaram maior chance de desenvolver hipercolesterolemia (OR =2,67; 95% IC: 1,265 - 5,657. O SNP rs1205 do gene CRP foi associado à hipercolesterolemia, podendo assim, aumentar o risco de problemas cardiovasculares na fase adulta.

  18. Rapid detection and subtyping of European swine influenza viruses in porcine clinical samples by haemagglutinin- and neuraminidase-specific tetra- and triplex real-time RT-PCRs

    DEFF Research Database (Denmark)

    Henritzi, Dinah; Zhao, Na; Starick, Elke

    2016-01-01

    Background A diversifying pool of mammalian-adapted influenza A viruses (IAV) with largely unknown zoonotic potential is maintained in domestic swine populations worldwide. The most recent human influenza pandemic in 2009 was caused by a virus with genes originating from IAV isolated from swine....... Swine influenza viruses (SIV) are widespread in European domestic pig populations and evolve dynamically. Knowledge regarding occurrence, spread and evolution of potentially zoonotic SIV in Europe is poorly understood. Objectives Efficient SIV surveillance programmes depend on sensitive and specific...... algorithm is proposed for the combined detection in clinical samples and subtyping of SIV strains currently circulating in Europe that is based on a generic, M-gene-specific influenza A virus RT-qPCR. In a second step, positive samples are examined by tetraplex HA- and triplex NA-specific RT...

  19. Importância da detecção das mutações no gene FLT3 e no gene NPM1 na leucemia mieloide aguda - Classificação da Organização Mundial de Saúde 2008 Importance of detecting FLT3 and NPM1 gene mutations in acute myeloid leukemia -World Health Organization Classification 2008

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    Marley Aparecida Licínio

    2010-01-01

    Full Text Available As leucemias mieloides agudas (LMA constituem um grupo de neoplasias malignas caracterizadas pela proliferação descontrolada de células hematopoéticas, decorrente de mutações que podem ocorrer em diferentes fases da diferenciação de células precursoras mieloides. Em 2008, a Organização Mundial da Saúde (OMS-2008 publicou uma nova classificação para neoplasias do sistema hematopoético e linfoide. De acordo com essa classificação, para um diagnóstico mais preciso e estratificação de prognóstico de pacientes com leucemias mieloides agudas, devem-se pesquisar mutações nos genes FLT3 e NPM1. Sabe-se que a presença de mutações no gene FLT3 é de prognóstico desfavorável e que as mutações no gene NPM1 do tipo A são de prognóstico favorável. Assim, nos países desenvolvidos, a análise das mutações no gene FLT3 e NPM1 tem sido considerada como um fator de prognóstico importante na decisão terapêutica em pacientes com diagnóstico de leucemias mieloides agudas. Considerando essas informações, é de extrema importância a análise das mutações no gene FLT3 (duplicação interna em tandem - DIT - e mutação pontual D835 e no gene NPM1 como marcadores moleculares para o diagnóstico, o prognóstico e a monitoração de doença residual mínima em pacientes com leucemias mieloides agudas.Acute myeloid leukemia (AML is a group of malignancies characterized by uncontrolled proliferation of hematopoietic cells resulting from mutations that occur at different stages in the differentiation of myeloid precursor cells. In 2008, the World Health Organization (WHO-2008 published a new classification for cancers of the hematopoietic and lymphoid system. According to this classification, FLT3 and NPM1 gene mutations should be investigated for a more precise diagnosis and prognostic stratification of AML patients. It is well known that the presence of FLT3 gene mutations is considered an unfavorable prognostic factor and type

  20. Expression of p190 BCR-ABL fusion gene in a patient with chronic myeloid leukemia Expressão do rearranjo gênico BCR-ABL com ponto de quebra na região menor do gene BCR em um paciente com leucemia mielóide crônica

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    P. V. B. Carvalho

    2003-01-01

    Full Text Available A minority of chronic myeloid leukemia cases have breakpoints in the minor cluster region (m-bcr of the BCR-ABL gene. We report on a patient with Ph-positive and m-bcr breakpoint at diagnosis. She was treated with hydroxyurea and interferon-alpha. Two years later, she developed a lymphoid blast crisis and died shortly after. We discuss herein the different forms of the BCR-ABL oncogene, its products, and the possible influence of them on the clinical outcome of patients with the disease.A leucemia mielóide crônica (LMC é uma doença mieloproliferativa clonal e caracteriza-se pela presença da translocação cromossômica entre os braços longos dos cromossomos 9 e 22, o denominado cromossomo Ph. Esta translocação determina a fusão dos genes BCR e ABL. Os diferentes pontos de quebra no gene BCR determinam a síntese de proteínas com diferentes pesos moleculares pelo gene BCR-ABL. Nós relatamos o caso de uma paciente portadora de LMC com ponto de quebra cromossômico na região menor do gene BCR. Foi tratada com hidroxiuréia e interferon alfa. Dois anos após o diagnóstico desenvolveu crise blástica linfóide e evoluiu rapidamente para o óbito. Nós discutimos nesta apresentação as diferentes formas do gene BCR-ABL e seus produtos e a possível influência dos mesmos na evolução clínica dos pacientes com a doença.

  1. Study of activity transcription factors C/EBPalpha in region - 53 to - 33 of promoter apolipoprotein B gene Estudo da atividade do fator de transcrição C/EBPalfa na região -53 a -33 do promoter do gene da apolipoproteina B

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    Kátia Cristina Dantas

    2006-09-01

    Full Text Available Apolipoprotein B (ApoB plays a major role in the regulation of cellular cholesterol homeostasis and pathogenesis of atherosclerosis. This protein acts as a ligand for the cellular recognition and catabolism of low density lipoprotein (LDL by the LDL receptor. Previous studies have shown that the expression of apoB in hepatic cells is regulated by the interaction of factors binding to enhancer elements in intron 2 and three elements designated III, IV and V. These elements lie within regions respectively -86 to -62, -72 to -53 and -53 to -33 from the ApoB promoter. In this study, we have suggested that transcription factor C/EBPalpha, which binds to the -53 to -33 region of the apoB, interacts with the HNF-4 synergistic complex and C/EBPalpha factors within -86 to -53 and may contribute to increase transcription of the ApoB gene.A apolipoproteina B (apoB tem um importante papel na regulação na homeostasia celular, do colesterol e na patogênese da aterosclerose. Esta proteína age como ligante para o reconhecimento e catabolismo lipoproteínas de baixa densidade (LBD através do receptor de LDL. Estudos anteriores mostraram que a expressão do gene da apolipoproteína B (APOB em células hepáticas é regulada pela interação de fatores ligados ao elemento enhancer no intron 2, e em 3 elementos denominados de III, IV e V localizados nas regiões -86 a -62, -72 a -53 e -53 a -33 , respectivamente, do promotor do gene da APOB. Neste trabalho, nós sugerimos que o fator de transcrição C/EBPalfa ligado a região -53 a -33 da APOB interage com o complexo HNF-4 e C/EBPalfa localizado dentro da região -86 a -53 do APO B e contribui para aumentar a transcrição do gene APOB.

  2. Evaluation of FcγRIIIB-NA1/NA2 Polymorphism in Visceral Leishmaniasis.

    Science.gov (United States)

    Abasi, Mohammad; Lotfi, Pegah; Bazmani, Ahad; Matini, Mohamad; Hajilooi, Mehrdad

    2014-04-01

    Several lines of evidence demonstrating that innate and adaptive immunity play important roles in the defense against visceral leishmaniasis (VL). A polymorphism within the FcγRIIIB gene can lead to the expression of three variants of NA1, NA2, and the combined one (NA1/NA2) which alters affinity of IgG to its receptor. The main aim of this study was to evaluate the FcγRIIIB-NA1/NA2 polymorphism in the FcγRIIIB gene of VL patients in comparison to healthy controls. In this cross-sectional study, three groups; 54 seropositive patients with clinical presentation of VL (group 1), 104 seropositive patients without clinical presentation (group 2), and 104 healthy controls (group 3) were evaluated with respect to the FcγRIIIB-NA1/NA2 polymorphism using a PCR-SSP method. The titration of anti-leishmania antibodies was analyzed using an immunoflorescence technique. Our results indicated that polymorphisms within the FcγRIIIB gene (that lead to the expression of the NA1/NA2 isoforms) are significantly associated with VL. The results demonstrated that the genotype heterozygotic for FcγRIIIB-NA1/NA2 expression was significantly increased in VL patients, group 1 when compared to groups 2 and 3. Conversely, there is a decrease in homozygous NA1 and NA2 genotypes in VL patients; however, the overall frequency of NA1 and NA2 alleles appear similar across the three cohorts examined. According to our results, it is likely that the increased frequency of the FcγRIIIB-NA1/NA2 genotype is associated with impaired immune responses against VL and its subsequent clearance from the patient.

  3. Filament-producing mutants of influenza A/Puerto Rico/8/1934 (H1N1 virus have higher neuraminidase activities than the spherical wild-type.

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    Jill Seladi-Schulman

    Full Text Available Influenza virus exhibits two morphologies - spherical and filamentous. Strains that have been grown extensively in laboratory substrates are comprised predominantly of spherical virions while clinical or low passage isolates produce a mixture of spheres and filamentous virions of varying lengths. The filamentous morphology can be lost upon continued passage in embryonated chicken eggs, a common laboratory substrate for influenza viruses. The fact that the filamentous morphology is maintained in nature but lost in favor of a spherical morphology in ovo suggests that filaments confer a selective advantage within the infected host that is not necessary for growth in laboratory substrates. Indeed, we have recently shown that filament-producing variant viruses are selected upon passage of the spherical laboratory strain A/Puerto Rico/8/1934 (H1N1 [PR8] in guinea pigs. Toward determining the nature of the selective advantage conferred by filaments, we sought to identify functional differences between spherical and filamentous particles. We compared the wild-type PR8 virus to two previously characterized recombinant PR8 viruses in which single point mutations within M1 confer a filamentous morphology. Our results indicate that these filamentous PR8 mutants have higher neuraminidase activities than the spherical PR8 virus. Conversely, no differences were observed in HAU:PFU or HAU:RNA ratios, binding avidity, sensitivity to immune serum in hemagglutination inhibition assays, or virion stability at elevated temperatures. Based on these results, we propose that the pleomorphic nature of influenza virus particles is important for the optimization of neuraminidase functions in vivo.

  4. Cloned cDNA of A/swine/Iowa/15/1930 internal genes as a candidate backbone for reverse genetics vaccine against influenza A viruses.

    Science.gov (United States)

    Lekcharoensuk, Porntippa; Wiriyarat, Witthawat; Petcharat, Nantawan; Lekcharoensuk, Chalermpol; Auewarakul, Prasert; Richt, Juergen A

    2012-02-14

    Reverse genetics viruses for influenza vaccine production usually utilize the internal genes of the egg-adapted A/Puerto Rico/8/34 (PR8) strain. This egg-adapted strain provides high production yield in embryonated eggs but does not necessarily give the best yield in mammalian cell culture. In order to generate a reverse genetics viral backbone that is well-adapted to high growth in mammalian cell culture, a swine influenza isolate A/swine/Iowa/15/30 (H1N1) (rg1930) that was shown to give high yield in Madin-Darby canine kidney (MDCK) cells was used as the internal gene donor for reverse genetics plasmids. In this report, the internal genes from rg1930 were used for construction of reverse genetics viruses carrying a cleavage site-modified hemagglutinin (HA) gene and neuraminidase (NA) gene from a highly pathogenic H5N1 virus. The resulting virus (rg1930H5N1) was low pathogenic in vivo. Inactivated rg1930H5N1 vaccine completely protected chickens from morbidity and mortality after challenge with highly pathogenic H5N1. Protective immunity was obtained when chickens were immunized with an inactivated vaccine consisting of at least 2(9) HA units of the rg1930H5N1 virus. In comparison to the PR8-based reverse genetics viruses carrying the same HA and NA genes from an H5N1 virus, rg1930 based viruses yielded higher viral titers in MDCK and Vero cells. In addition, the reverse genetics derived H3N2 and H5N2 viruses with the rg1930 backbone replicated in MDCK cells better than the cognate viruses with the rgPR8 backbone. It is concluded that this newly established reverse genetics backbone system could serve as a candidate for a master donor strain for development of inactivated influenza vaccines in cell-based systems. Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. Perfil de expressÃo e anÃlise filogenÃtica dos genes da proteÃna desacopladora mitocondrial durante o desenvolvimento e estresse em soja [Glycine max (L.) Merr.

    OpenAIRE

    AntÃnio Edson Rocha Oliveira

    2015-01-01

    Diversos estudos tÃm evidenciado que a principal funÃÃo da proteÃna desacopladora mitocondrial de plantas (pUCP) està relacionada a regulaÃÃo de espÃcies reativas de oxigÃnio (EROs). AnÃlises in silico sugerem a existÃncia de famÃlias multigÃnicas para a codificaÃÃo de pUCPs, porÃm novos estudos ainda sÃo necessÃrios para estabelecer o perfil de expressÃo gÃnica das pUCPs, assim como a quantidade de genes em cada espÃcie e suas relaÃÃes filogenÃticas. O presente trabalho teve como objetivo ca...

  6. Impact of engineered Streptococcus thermophilus trains overexpressing glyA gene on folic acid and acetaldehyde production in fermented milk Impacto de linhagens de Streptococcus thermophilus com aumento da expressão do gene glyA na produção de ácido folico e acetaldeído em leite fermentado

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    Ana Carolina Sampaio Dória Chaves

    2003-11-01

    Full Text Available The typical yogurt flavor is caused by acetaldehyde produced through many different pathways by the yogurt starter bacteria L. bulgaricus and S. thermophilus. The attention was focused on one specific reaction for acetaldehyde and folic acid formation catalyzed by serine hydroxymethyltransferase (SHMT, encoded by the glyA gene. In S. thermophilus, this enzyme SHMT also plays the typical role of the enzyme threonine aldolase (TA that is the interconvertion of threonine into glycine and acetaldehyde. The behavior of engineered S. thermophilus strains in milk fermentation is described, folic acid and acetaldehyde production were measured and pH and counts were followed. The engineered S. thermophilus strains StA2305 and StB2305, have the glyA gene (encoding the enzyme serine hydroxymethyltransferase overexpressed. These engineered strains showed normal growth in milk when it was supplemented with Casitione. When they were used in milk fermentation it was observed an increase in folic acid and in acetaldehyde production by StA2305 and for StB2305 it was noticed a significative increase in folic acid formation.O acetaldeído, responsável pelo sabor e aroma característicos de iogurte, é produzido por diferentes vias metabólicas pelas bactérias lácticas: Streptococcus thermophilus (S. thermophilus e Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus. Neste trabalho, a atenção foi focada especificamente na reação para a formação de acetaldeído e de ácido fólico, catalisada pela enzima serina hidroximetil transferase (SHMT, codificada pelo gene glyA. A enzima SHMT catalisa diversas reações e, no caso da bactéria S. thermophilus, ela exerce também a atividade característica da enzima treonina aldolase (TA, definida como a interconversão do aminoácido treonina em glicina e acetaldeído. Foram construídas linhagens de S. thermophilus (StA2305 e StB2305 com super expressão do gene glyA. Estas linhagens modificadas apresentaram

  7. Environmental exposures and gene regulation in disease etiology Exposição ambiental e regulação genética na etiologia de doenças

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    Thea M. Edwards

    2008-02-01

    ônios, produtos de nutrição e comportamentos podem mudar a expressão genética por meio de uma quantidade enorme de mecanismos regulatórios dos genes. Ademais, mudanças quimicamente induzidas na regulação do gene estão associadas a enfermidades graves e complexas, como é o caso do câncer, diabetes, infertilidade, doenças respiratórias, alergias e problemas neurodegenerativos como o mal de Parkinson e Alzheimer. Os estudos revistos indicam que uma predisposição genética para determinada doença é melhor prevista no contexto das exposições ambientais. E os mecanismos genéticos examinados nesses estudos oferecem novos caminhos para pesquisas sobre avaliação de risco.

  8. Single gene deletions of mrpA to mrpG and mrpE point mutations affect activity of the Mrp Na+/H+ antiporter of alkaliphilic Bacillus and formation of hetero-oligomeric Mrp complexes.

    Science.gov (United States)

    Morino, Masato; Natsui, Shinsuke; Swartz, Talia H; Krulwich, Terry A; Ito, Masahiro

    2008-06-01

    Mrp antiporters catalyze secondary Na(+)(Li(+))/H(+) antiport and/or K(+)/H(+) antiport that is physiologically important in diverse bacteria. An additional capacity for anion flux has been observed for a few systems. Mrp is unique among antiporters in that it requires all six or seven hydrophobic gene products (MrpA to MrpG) of the mrp operon for full antiporter activity, but MrpE has been reported to be dispensable. Here, the membrane complexes formed by Mrp proteins were examined using a cloned mrp operon from alkaliphilic Bacillus pseudofirmus OF4. The operon was engineered so that the seven Mrp proteins could be detected in single samples. Membrane extracts of an antiporter-deficient Escherichia coli strain expressing this construct were analyzed by blue native-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Mrp complexes of two sizes were identified containing all seven Mrp proteins. Studies of the single nonpolar mrp gene deletions in the construct showed that a subcomplex of MrpA, MrpB, MrpC, and MrpD was formed in the absence of MrpE, MrpF, or MrpG. By contrast, MrpE, MrpF, and MrpG were not observed in membranes lacking MrpA, MrpB, MrpC, or MrpD. Although MrpA and MrpD have been hypothesized to be the antiporter proteins, the MrpA-to-D complex was inactive. Every Mrp protein was required for an activity level near that of the wild-type Na(+)/H(+) antiporter, but a very low activity level was observed in the absence of MrpE. The introduction of an MrpE(P114G) mutation into the full Mrp complex led to antiport activity with a greatly increased apparent K(m) value for Na(+). The results suggested that interactions among the proteins of heterooligomeric Mrp complexes strongly impact antiporter properties.

  9. Pesquisa de marcadores para os genes da cadeia pesada da beta-miosina cardíaca e da proteína C de ligação à miosina em familiares de pacientes com cardiomiopatia hipertrófica Research of markers for the genes of the heavy chain of cardiac beta-myosin and myosin binding protein C in relatives of patients with hypertrophic cardiomyopathy

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    Adriana Paula Tirone

    2005-06-01

    Full Text Available OBJETIVO: Estudar os marcadores moleculares para os genes da cadeia pesada da beta-miosina cardíaca e da proteína-C de ligação à miosina em familiares de portadores de cardiomiopatia hipertrófica. MÉTODOS: Foram estudadas 12 famílias que realizaram anamnese, exame físico, eletrocardiograma, ecocardiograma e coleta de sangue para o estudo genético através da reação em cadeia da polimerasse. RESULTADOS: Dos 227 familiares 25% eram acometidos, sendo 51% do sexo masculino com idade média de 35±19 (2 a 95 anos. A análise genética mostrou ligação com o gene da b-miosina cardíaca em uma família e, em outra, ligação com o gene da proteína C de ligação à miosina. Em cinco famílias foram excluídas ligações com os dois genes; em duas, a ligação com o gene da proteína C de ligação à miosina, porém para o gene da b-miosina os resultados foram inconclusivos; em duas famílias os resultados foram inconclusivos para os dois genes e em uma foi excluída ligação para o gene da b-miosina mas ficou inconclusivo para o gene da proteína C de ligação à miosina. CONCLUSÃO: Em nosso meio, talvez predominem outros genes que não aqueles descritos na literatura, ou que existam outras diferenças genéticas relacionadas com a origem de nossa população e/ou fatores ambientais.OBJECTIVE: To study the molecular markers for the genes of the heavy chain of cardiac beta-myosin and the myosin binding protein C in relatives of carriers of hypertrophic cardiomyopathy. METHODS: Twelve families who had anamnesis, physical exam, electrocardiogram, echocardiogram and blood collection for the genetic study through the chain reaction of polymerase. RESULTS: From the 227 relatives, 25% were ill-taken, with 51% men, with an average age of 35±19 (2 to 95 years old. The genetic analysis showed a connection with the gene of the cardiac b-myosin in a family and, in another, a connection with the gene of the myosin-binding protein C. In five

  10. ANÁLISE DA EXPRESSÃO DOS GENES SERPINA1 E TFF3 NA PREDIÇÃO DO ENVOLVIMENTO DE LINFONODOS EM NEOPLASIAS MAMÁRIAS

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    Raissa Monteiro da Silva

    2017-02-01

    Full Text Available O status dos linfonodos axilares é o fator prognóstico mais informativo no tratamento das pacientes com câncer de mama. Assim, a identificação de marcadores moleculares no tumor primário que possa permitir uma classificação mais precisa das pacientes em relação à necessidade, é extremamente importante para uma conduta clínica mais adequada. O objetivo deste estudo foi determinar se os genes SERPINA1 e TFF3 são marcadores preditivos em câncer de mama pela análise de expressão gênica de RT-qPCR. Para isto, foi comparado o grupo de tumores primários com envolvimento de linfonodos e tumores primários sem linfonodos acometidos, além da análise dos linfonodos correspondentes. Nossos resultados demonstraram que ambos os genes analisados não possuem papel preditivo relacionado ao desenvolvimento destes carcinomas.

  11. Impact of a large deletion in the neuraminidase protein identified in a laninamivir-selected influenza A/Brisbane/10/2007 (H3N2) variant on viral fitness in vitro and in ferrets.

    Science.gov (United States)

    Ann, Julie; Abed, Yacine; Beaulieu, Edith; Bouhy, Xavier; Joly, Marie-Hélène; Dubé, Karen; Carbonneau, Julie; Hamelin, Marie-Eve; Mallett, Corey; Boivin, Guy

    2016-03-01

    Viral fitness of a laninamivir-selected influenza A/Brisbane/10/2007-like (H3N2) isolate (LRVp9) containing a 237-amino acid neuraminidase deletion and a P194L hemagglutinin mutation was evaluated in vitro and in ferrets. LRVp9 and the wild-type (WT) virus showed comparable replication kinetics in MDCK-ST6GalI cells. Cultured virus was recovered between days 2 and 5 post-infection in nasal washes (NW) from the 4 WT-infected ferrets whereas no virus was recovered from the LRVp9-infected animals. There was a ≥1 log reduction in viral RNA copies/μl of NW for LRVp9 compared to WT at most time points. The large neuraminidase deletion compromises viral infectivity in vivo. © 2015 The Authors. Influenza and Other Respiratory Viruses Published by John Wiley & Sons Ltd.

  12. Estudo genético do gene p16 pela técnica de PCR-SSCP e expressão de proteína p16 em melanomas de mucosa oral e melanomas cutâneos Genetic analysis of p16 gene by PCR-SSCP technique and protein p16 expression in oral mucosa and skin melanomas

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    Ricardo Hsieh

    2006-10-01

    Full Text Available FUNDAMENTOS: A deleção e mutação do gene CDKN2a que codifica um inibidor específico da ciclina dependente de quinase 4, a proteína p16, têm sido implicadas na tumorigênese do melanoma cutâneo. Entretanto, pouco se conhece sobre essas alterações genéticas em melanomas de mucosa oral. OBJETIVOS: Verificar a presença de alterações no gene p16 e sua expressão protéica em melanomas esporádicos orais e cutâneos. MATERIAL E MÉTODOS: Avaliaram-se 36 espécimes de melanoma primário (sete orais e 29 cutâneos. Analisaram-se três exons do gene p16, pela técnica da reação em cadeia da polimerase/polimorfismo conformacional de fita simples do DNA.Verificou-se a expressão tecidual de proteína p16 por técnica imuno-histoquímica. Relacionaram-se os resultados com a espessura dos melanomas cutâneos. RESULTADOS: Cinco dos sete melanomas orais e 17 dos 29 melanomas cutâneos apresentaram indício de alteração no gene p16. Alterações do exon 2 foram as mais freqüentes, sendo 19 casos nos produtos obtidos com o mesmo iniciador. Observou-se expressão tecidual de p16 em apenas um melanoma oral, em 10/13 (76,9% casos de melanoma cutâneo de espessura até 1mm e em sete de oito (87,5% casos de espessura superior a 1mm. CONCLUSÃO: A freqüência de indícios de alteração na análise genética de p16 nos melanomas de mucosa oral foi de 71,42% e de 58,6% nos cutâneos. É possível sugerir a participação de alterações do gene p16 na patogenia do melanoma esporádico de mucosa oral. Não houve relação da sugestão de alteração genética do gene p16 e de sua expressão tecidual com a espessura dos melanomas cutâneos de diferentes subtipos histológicos.BACKGROUND: Deletion and mutation of gene CDKN2a, which encodes a specific inhibitor of cyclin-dependent kinase 4 (CDK4, the protein p16, has been regarded as related to cutaneous melanoma tumorigenesis. However, little is known about those alterations in oral mucosa melanomas

  13. Evaluation of the zoonotic potential of a novel reassortant H1N2 swine influenza virus with gene constellation derived from multiple viral sources.

    Science.gov (United States)

    Lee, Jee Hoon; Pascua, Philippe Noriel Q; Decano, Arun G; Kim, Se Mi; Park, Su-Jin; Kwon, Hyeok-Il; Kim, Eun-Ha; Kim, Young-Il; Kim, HyongKyu; Kim, Seok-Yong; Song, Min-Suk; Jang, Hyung-Kwan; Park, Bong Kyun; Choi, Young Ki

    2015-08-01

    In 2011-2012, contemporary North American-like H3N2 swine influenza viruses (SIVs) possessing the 2009 pandemic H1N1 matrix gene (H3N2pM-like virus) were detected in domestic pigs of South Korea where H1N2 SIV strains are endemic. More recently, we isolated novel reassortant H1N2 SIVs bearing the Eurasian avian-like swine H1-like hemagglutinin and Korean swine H1N2-like neuraminidase in the internal gene backbone of the H3N2pM-like virus. In the present study, we clearly provide evidence on the genetic origins of the novel H1N2 SIVs virus through genetic and phylogenetic analyses. In vitro studies demonstrated that, in comparison with a pre-existing 2012 Korean H1N2 SIV [A/swine/Korea/CY03-11/2012 (CY03-11/2012)], the 2013 novel reassortant H1N2 isolate [A/swine/Korea/CY0423/2013 (CY0423-12/2013)] replicated more efficiently in differentiated primary human bronchial epithelial cells. The CY0423-12/2013 virus induced higher viral titers than the CY03-11/2012 virus in the lungs and nasal turbinates of infected mice and nasal wash samples of ferrets. Moreover, the 2013 H1N2 reassortant, but not the intact 2012 H1N2 virus, was transmissible to naïve contact ferrets via respiratory-droplets. Noting that the viral precursors have the ability to infect humans, our findings highlight the potential threat of a novel reassortant H1N2 SIV to public health and underscore the need to further strengthen influenza surveillance strategies worldwide, including swine populations. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. AS IMPLICAÇÕES DO POLIMORFISMO GENÉTICO DO GENE GST NA PATOGÊNESE DO DIABETES MELLITUS TIPO 2

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    Angela Adamski da Silva Reis

    2011-01-01

    Full Text Available Type 2 Diabetes mellitus (T2DM is a disease characterized by chronic hyperglycemia associated with metabolic complications, clinical and social, resulting from the aging population, increasing urbanization and the adoption of unhealthy lifestyles such as sedentary lifestyles, poor diet and especially obesity. Complex diseases like diabetes, are determined by three main factors: the lifestyle, environmental exposure and genetic susceptibility. Thegenetic contribution is important evidence in the development of T2DM. Several polymorphisms of genes responsible for encoding enzymes involved in the biotransformation of xenobiotics are related to the development of atherosclerosis, cancer and type 2 diabetes. In the pathogenesis of T2DM, it is observed that oxidative stress contributes to decreased production and reduced ability of insulin to stimulate glucose uptake. In this sense, the polymorphism ofenzymes of xenobiotic metabolism phase II has been extensively studied in a wide variety of diseases, especially in determining genetic susceptibility to cancer and response to chemotherapy.

  15. Precise gene editing of chicken Na+/H+ exchange type 1 (chNHE1) confers resistance to avian leukosis virus subgroup J (ALV-J).

    Science.gov (United States)

    Lee, Hong Jo; Lee, Kyung Youn; Jung, Kyung Min; Park, Kyung Je; Lee, Ko On; Suh, Jeong-Yong; Yao, Yongxiu; Nair, Venugopal; Han, Jae Yong

    2017-12-01

    Avian leukosis virus subgroup J (ALV-J), first isolated in the late 1980s, has caused economic losses to the poultry industry in many countries. As all chicken lines studied to date are susceptible to ALV infection, there is enormous interest in developing resistant chicken lines. The ALV-J receptor, chicken Na+/H+ exchange 1 (chNHE1) and the critical amino acid sequences involved in viral attachment and entry have already been characterized. However, there are no reported attempts to induce resistance to the virus by targeted genome modification of the receptor sequences. In an attempt to induce resistance to ALV-J infection, we used clustered regularly interspaced short palindromic repeats (CRISPR)-associated (CRISPR/Cas9)-based genome editing approaches to modify critical residues of the chNHE1 receptor in chicken cells. The susceptibility of the modified cell lines to ALV-J infection was examined using enhanced green fluorescent protein (EGFP)-expressing marker viruses. We showed that modifying the chNHE1 receptor by artificially generating a premature stop codon induced absolute resistance to viral infection, with mutations of the tryptophan residue at position 38 (Trp38) being very critical. Single-stranded oligodeoxynucleotide (ssODN)-mediated targeted recombination of the Trp38 region revealed that deletions involving the Trp38 residue were most effective in conferring resistance to ALV-J. Moreover, protein structure analysis of the chNHE1 receptor sequence suggested that its intrinsically disordered region undergoes local conformational changes through genetic alteration. Collectively, these results demonstrate that targeted mutations on chNHE1 alter the susceptibility to ALV-J and the technique is expected to contribute to develop disease-resistant chicken lines. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Detecção dos genes codificantes da toxina CDT, e pesquisa de fatores que influenciam na produção de hemolisinas em amostras de Campylobacter jejuni de origem avícola

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    Michele M. Trindade

    2015-08-01

    Full Text Available Resumo: Membros termofílicos do gênero Campylobacter são reconhecidos como importantes enteropatógenos para o ser humano e animais. A grande diversidade ecológica destes micro-organismos em diferentes habitats tais como água, animais e alimentos predispõem ao aparecimento de novos fatores de virulência. Este trabalho teve por objetivo detectar os genes codificantes da Toxina Distensiva Citoletal (CDT por meio da técnica de PCR, pesquisar a atividade de hemolisinas e a influência de soluções quelantes e de íons nesta atividade. Foram utilizadas 45 amostras de Campylobacter jejuni de origem avícola para pesquisa de atividade hemolítica, cultivadas em Caldo Triptona de Soja (TSB. Após o crescimento bacteriano, as amostras foram semeadas em Ágar tríptico de soja (TSA contendo 5% de sangue de ovino. Para verificar a influência de agentes quelantes e solução de íons na atividade hemolítica, as amostras de C. jejuni foram cultivadas em TSB contendo separadamente os quelantes EDTA, ácido acético, soluções de íons CaCl2, MgCl2 e FeCl3, em atmosfera de microaerofilia. Quanto à atividade de hemolisina de C. jejuni em placas de TSA - sangue ovino foi possível observar que houve hemólise em 40% das amostras analisadas apenas com caldo TSB. Somente o ácido acético apresentou ação quelante sobre a atividade de hemolisinas em amostras de C. jejuni semeadas em placas de TSA - sangue ovino. Para detecção dos genes cdtA, cdtB e cdtC através da técnica da Reação em Cadeia da Polimerase (PCR foram utilizadas 119 amostras de C. jejuni de origem avícola. Foi possível observar que 37,8% possuíam o perfil de genes cdtABC. Os resultados demonstraram em amostras avícolas a presença de cepas de C. jejuni com potencial virulento, devido à presença dos genes da toxina CDT e potencial hemolítico, que apresentou ação reduzida in vitro com ácido acético.

  17. Caracterização de genes diferencialmente expressos na interação entre maçã 'Fuji' e Botryosphaeria dothidea Characterization of differently expressed genes during the interaction between 'Fuji' apples and Botryosphaeria dothidea

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    Adriana Regina Corrent

    2009-03-01

    Full Text Available O objetivo deste estudo foi investigar a interação entre o fungo Botryosphaeria dothidea e maçãs cv. Fuji por meio da técnica de Differential Display RT-PCR. O cDNA de frutos infectados e não infectados pelo fungo foi amplificado com uma combinação de 15 oligonucleotídeos iniciadores. Foram isolados 400 fragmentos de cDNA diferencialmente expressos, dos quais 120 foram sequenciados e comparados com sequências disponíveis no GenBank, por meio do programa BLASTX. As sequências obtidas foram similares à metalotioninas, profilina alergênica, proteína de resistência e fosfatase.The aim of this study was to investigate the interaction between the Botryosphaeria dothidea fungi and 'Fuji' apples using the Differential Display RT-PCR technique. The cDNA of infected and non infected fruits by the fungus was amplified using a combination of 15 primers. Four hundred fragments of differentially expressed cDNA were isolated, and 120 of them were sequenced and compared with available sequences at GenBank database, using BLASTX software. Obtained sequences were similar to methalothionines, an allergenic profiline, a resistance linked protein, and a phosphatase.

  18. Wheat germ cell-free system-based production of hemagglutinin-neuraminidase protein of human parainfluenza virus type 3: generation and characterization of monoclonal antibody

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    Satoko eMatsunaga

    2014-05-01

    Full Text Available Human parainfluenza virus 3 (HPIV3 commonly causes respiratory disorders in infants and young children. Monoclonal antibodies (MAbs have been produced to several components of HPIV3 and commercially available. However, the utility of these antibodies for several immunological and proteomic assays for understanding the nature of HPIV3 infection remain to be characterized. Herein, we report the development and characterization of monoclonal antibodies against hemagglutinin-neuraminidase (HN of HPIV3. A recombinant full-length HPIV3-HN was successfully synthesized using the wheat-germ cell-free protein production system. After immunization and cell fusion, 36 mouse hybridomas producing MAbs to HPIV3-HN were established. The MAbs obtained were fully characterized using ELISA, immunoblotting and immunofluorescent analyses. Of the MAbs tested, single clone was found to be applicable in both flow cytometry and immunoprecipitation procedures. By utilizing the antibody, we newly identified HPIV3-HN binding host proteins via immunoprecipitation-based mass spectrometry analysis. This study provides the availability of our newly-developed MAbs as a valuable tool for the study of HPIV3 infection as well as the several diagnostic tests of this virus.

  19. Combined quantum mechanics/molecular mechanics (QM/MM) simulations for protein-ligand complexes: free energies of binding of water molecules in influenza neuraminidase.

    Science.gov (United States)

    Woods, Christopher J; Shaw, Katherine E; Mulholland, Adrian J

    2015-01-22

    The applicability of combined quantum mechanics/molecular mechanics (QM/MM) methods for the calculation of absolute binding free energies of conserved water molecules in protein/ligand complexes is demonstrated. Here, we apply QM/MM Monte Carlo simulations to investigate binding of water molecules to influenza neuraminidase. We investigate five different complexes, including those with the drugs oseltamivir and peramivir. We investigate water molecules in two different environments, one more hydrophobic and one hydrophilic. We calculate the free-energy change for perturbation of a QM to MM representation of the bound water molecule. The calculations are performed at the BLYP/aVDZ (QM) and TIP4P (MM) levels of theory, which we have previously demonstrated to be consistent with one another for QM/MM modeling. The results show that the QM to MM perturbation is significant in both environments (greater than 1 kcal mol(-1)) and larger in the more hydrophilic site. Comparison with the same perturbation in bulk water shows that this makes a contribution to binding. The results quantify how electronic polarization differences in different environments affect binding affinity and also demonstrate that extensive, converged QM/MM free-energy simulations, with good levels of QM theory, are now practical for protein/ligand complexes.

  20. The in vivo efficacy of neuraminidase inhibitors cannot be determined from the decay rates of influenza viral titers observed in treated patients

    Science.gov (United States)

    Palmer, John; Dobrovolny, Hana M.; Beauchemin, Catherine A. A.

    2017-01-01

    Antiviral therapy is a first line of defence against new influenza strains. Current pandemic preparations involve stock- piling oseltamivir, an oral neuraminidase inhibitor (NAI), so rapidly determining the effectiveness of NAIs against new viral strains is vital for deciding how to use the stockpile. Previous studies have shown that it is possible to extract the drug efficacy of antivirals from the viral decay rate of chronic infections. In the present work, we use a nonlinear mathematical model representing the course of an influenza infection to explore the possibility of extracting NAI drug efficacy using only the observed viral titer decay rates seen in patients. We first show that the effect of a time-varying antiviral concentration can be accurately approximated by a constant efficacy. We derive a relationship relating the true treatment dose and time elapsed between doses to the constant drug dose required to approximate the time- varying dose. Unfortunately, even with the simplification of a constant drug efficacy, we show that the viral decay rate depends not just on drug efficacy, but also on several viral infection parameters, such as infection and production rate, so that it is not possible to extract drug efficacy from viral decay rate alone.

  1. Production of an enzymatically active and immunogenic form of ectodomain of Porcine rubulavirus hemagglutinin-neuraminidase in the yeast Pichia pastoris.

    Science.gov (United States)

    Cerriteño-Sánchez, José Luis; Santos-López, Gerardo; Rosas-Murrieta, Nora Hilda; Reyes-Leyva, Julio; Cuevas-Romero, Sandra; Herrera-Camacho, Irma

    2016-04-10

    Blue-eye disease (BED) of swine is a viral disease endemic in Mexico. The etiological agent is a paramyxovirus classified as Porcine rubulavirus (PoRV-LPMV), which exhibits in its envelope the hemagglutinin-neuraminidase (HN) glycoprotein, the most immunogenic and a major target for vaccine development. We report in this study the obtaining of ectodomain of PoRV HN (eHN) through the Pichia pastoris expression system. The expression vector (pPICZαB-HN) was integrated by displacement into the yeast chromosome and resulted in a Mut(+) phenotype. Expressed eHN in the P. pastoris X33 strain was recovered from cell-free medium, featuring up to 67 nmol/min/mg after 6 days of expression. eHN was recognized by the serum of infected pigs with strains currently circulating in the Mexican Bajio region. eHN induces antibodies in mice after 28 days of immunization with specific recognition in ELISA test. These antibodies were able to inhibit >80% replication by viral neutralization assays in cell culture. These studies show the obtaining of a protein with similar characteristics to the native HN and which may be a candidate to propose a vaccine or to use the antigen in a serologic diagnostic test. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Reassortment between Avian H5N1 and Human Influenza Viruses Is Mainly Restricted to the Matrix and Neuraminidase Gene Segments

    NARCIS (Netherlands)

    E.J.A. Schrauwen (Eefje); T.M. Bestebroer (Theo); G.F. Rimmelzwaan (Guus); A.D.M.E. Osterhaus (Albert); R.A.M. Fouchier (Ron); S. Herfst (Sander)

    2013-01-01

    textabstractHighly pathogenic avian influenza H5N1 viruses have devastated the poultry industry in many countries of the eastern hemisphere. Occasionally H5N1 viruses cross the species barrier and infect humans, sometimes with a severe clinical outcome. When this happens, there is a chance of

  3. In vitro susceptibility to quinine and microsatellite variations of the Plasmodium falciparum Na+/H+ exchanger (Pfnhe-1 gene: the absence of association in clinical isolates from the Republic of Congo

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    Rogier Christophe

    2011-02-01

    Full Text Available Abstract Background Quinine is still recommended as an effective therapy for severe cases of Plasmodium falciparum malaria, but the parasite has developed resistance to the drug in some cases. Investigations into the genetic basis for quinine resistance (QNR suggest that QNR is complex and involves several genes, with either an additive or a pairwise effect. The results obtained when assessing one of these genes, the plasmodial Na+/H+ exchanger, Pfnhe-1, were found to depend upon the geographic origin of the parasite strain. Most of the associations identified have been made in Asian strains; in contrast, in African strains, the influence of Pfnhe on QNR is not apparent. However, a recent study carried out in Kenya did show a significant association between a Pfnhe polymorphism and QNR. As genetic differences may exist across the African continent, more field data are needed to determine if this association exists in other African regions. In the present study, association between Pfnhe and QNR is investigated in a series of isolates from central Africa. Methods The sequence analysis of the polymorphisms at the Pfnhe-1 ms4760 microsatellite and the evaluation of in vitro quinine susceptibility (by isotopic assay were conducted in 74 P. falciparum isolates from the Republic of Congo. Results Polymorphisms in the number of DNNND or NHNDNHNNDDD repeats in the Pfnhe-1 ms4760 microsatellite were not associated with quinine susceptibility. Conclusions The polymorphism in the microsatellite ms4760 in Pfnhe-1 that cannot be used to monitor quinine response in the regions of the Republic of Congo, where the isolates came from. This finding suggests that there exists a genetic background associated with geographic area for the association that will prevent the use of Pfnhe as a molecular marker for QNR. The contribution of Pfnhe to the in vitro response to quinine remains to be assessed in other regions, including in countries with different levels of

  4. A novel missense mutation pattern of the GCH1 gene in dopa-responsive dystonia Novo padrão de mutação missense no gene GCH1 na distonia dopa-responsiva

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    Rosana H. Scola

    2007-12-01

    Full Text Available Dopa-responsive dystonia (DRD is an inherited metabolic disorder now classified as DYT5 with two different biochemical defects: autosomal dominant GTP cyclohydrolase 1 (GCH1 deficiency or autosomal recessive tyrosine hydroxylase deficiency. We report the case of a 10-years-old girl with progressive generalized dystonia and gait disorder who presented dramatic response to levodopa. The phenylalanine to tyrosine ratio was significantly higher after phenylalanine loading test. This condition had two different heterozygous mutations in the GCH1 gene: the previously reported P23L mutation and a new Q182E mutation. The characteristics of the DRD and the molecular genetic findings are discussed.Distonia dopa-responsiva (DRD, classificada como DYT5, é um erro inato do metabolismo que pode ser causado por dois diferentes tipos de defeito bioquímico: deficiência de GTP ciclo-hidrolase 1 (GCH1 (autossômica dominante ou de tirosina hidroxilase (autossômica recessiva. Descrevemos o caso de menina de 10 anos com distonia generalizada progressiva e alteração da marcha com importante melhora após uso de levodopa. A relação fenilalanina/tirosina estava aumentada após teste de sobrecarga com fenilalanina. O estudo molecular mostrou que o paciente apresenta uma combinação hererozigótica de mutação no gene GCH1: a já conhecida mutação P23L e uma nova mutação Q182E. Discutem-se as características da DRD e as alterações genéticas possíveis.

  5. Isolation and characterization of ScGluD2, a new sugarcane beta-1,3-glucanase D family gene induced by Sporisorium scitamineum, ABA, H2O2, NaCl, and CdCl2 stresses

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    Yachun Su

    2016-09-01

    Full Text Available Beta-1,3-glucanases (EC 3.2.1.39, commonly known as pathogenesis-related (PR proteins, play an important role not only in plant defense against fungal pathogens but also in plant physiological and developmental processes. However, only a limited number of sugarcane beta-1,3-glucanase genes have been isolated. In the present study, we identified and characterized a new beta-1,3-glucanase gene ScGluD2 (GenBank Acc No. KF664181 from sugarcane. An X8 domain was present at the C terminal region of ScGluD2, suggesting beta-1,3-glucan-binding function. Phylogenetic analysis showed that the predicted ScGluD2 protein was classified into subfamily D beta-1,3-glucanase. Localization of the ScGluD2 protein in the plasma membrane was determined by tagging it with green fluorescent protein. The expression of ScGluD2 was more up-regulated in sugarcane smut-resistant cultivars in the early stage (1 d or 3 d than in the susceptible ones after being challenged by the smut pathogen, revealing that ScGluD2 may be involved in defense against the invasion of Sporisorium scitamineum. Transient overexpression of ScGluD2 in Nicotiana benthamiana leaves induced a defense response and exhibited antimicrobial action on the tobacco pathogens Pseudomonas solanacearum and Botrytis cinerea, further demonstrating that ScGluD2 was related to the resistance to plant pathogens. However, the transcripts of ScGluD2 partially increased (12 h under NaCl stress, and were steadily up-regulated from 6 h to 24 h upon ABA, H2O2, and CdCl2 treatments, suggesting that ABA may be a signal molecule regulating oxidative stress and play a role in the salt and heavy metal stress-induced stimulation of ScGluD2 transcripts. Taken together, ScGluD2, a novel member of subfamily D beta-1,3-glucanase, was a stress-related gene of sugarcane involved in plant defense against smut pathogen attack and salt and heavy metal stresses.

  6. Efeito do genótipo halotano e de diferentes sistemas de produção no desempenho produtivo e na qualidade da carcaça suína Effect of the halothane genes and rearing systems on growth performance and pig carcass quality

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    Ana Maria Bridi

    2003-08-01

    Full Text Available O objetivo deste trabalho foi verificar o efeito dos genótipos halotano homozigoto dominante e heterozigoto e de três sistemas de criação (confinado sobre piso de cimento, confinado sobre cama de maravalha e ao ar livre no desempenho produtivo e na qualidade da carcaça e da carne suína. Foram utilizados 96 suínos machos castrados selecionados pelo exame de DNA genômico no sangue utilizando-se a técnica de reação em cadeia da polimerase (PCR para amplificar a região do receptor rianodina. A região amplificada foi clivada pela técnica do polimorfismo do comprimento dos fragmentos de restrição (RFLP. Nas fases de crescimento e terminação, avaliaram-se ganho de peso, consumo de ração e conversão alimentar. Após o abate, as carcaças foram pesadas e tipificadas eletronicamente. Vinte e quatro horas após o abate, foram separados e pesados o pernil e o carrê e medida a área de olho de lombo. A presença do gene halotano não afetou o desempenho dos suínos, mas aumentou a porcentagem de carne magra e diminuiu a quantidade de gordura na carcaça. Suínos criados no sistema intensivo ao ar livre apresentaram menor ganho de peso e pior conversão alimentar. No sistema intensivo de criação confinado sobre cama, os suínos produziram carcaças com menor quantidade de carne magra e maior quantidade de gordura. Não houve efeito significativo da interação genótipo halotano e sistema de criação para as características avaliadas.The objective of this study was to evaluate the effect of the halothane genotypes (heterozygous and dominant homozygous and intensive rearing systems (indoor, wood shavings bedding and outdoor on growth performance, carcass and meat yield. Ninety six castrated male pigs were used for the trial. Identification of the halothane genotype was determined in blood samples using the DNA-test, based on the polymerase chain reaction (PCR for amplification of the critical region of the ryanodine receptor and

  7. Avaliação da taxa de metilação do DNA de leucócitos na região promotora dos genes IFNγ, Serpin B5 e Stratifin durante o período gestacional e sua relação com o metabolismo das vitaminas e metabólitos

    OpenAIRE

    Thaiomara Alves Silva

    2010-01-01

    A metilação do DNA é uma alteração epigenética que atua na regulação da expressão gênica. A deficiência de vitaminas (cobalamina, B6 e folato) pode interferir na taxa de metilação. O efeito da deficiência destas vitaminas foi determinado em estudos com cultura de células e em animais. No entanto, são raros os estudos realizados com seres humanos. Os objetivos deste trabalho foram: avaliar a taxa de metilação do DNA de leucócitos na região promotora dos genes Interferon gama (IFNγ), Serpi...

  8. 6SLN-lipo PGA specifically catches (coats) human influenza virus and synergizes neuraminidase-targeting drugs for human influenza therapeutic potential.

    Science.gov (United States)

    Sriwilaijaroen, Nongluk; Suzuki, Katsuhiko; Takashita, Emi; Hiramatsu, Hiroaki; Kanie, Osamu; Suzuki, Yasuo

    2015-10-01

    The purpose of this study was to develop a new compound to overcome influenza epidemics and pandemics as well as drug resistance. We synthesized a new compound carrying: (i) Neu5Acα2-6Galβ1-4GlcNAc (6SLN) for targeting immutable haemagglutinins (HAs) unless switched from human-type receptor preference; (ii) an acyl chain (lipo) for locking the compound with the viral HA via hydrophobic interactions; and (iii) a flexible poly-α-L-glutamic acid (PGA) for enhancing the compound solubility and for coating the viral surface, precluding accessibility of the PGA-coated virus to the negatively charged sialic acid on the host cell surface. 6SLN-lipo PGA appears to subvert binding of pandemic H1 and seasonal H3 HAs to receptors, as assessed by using guinea pig erythrocytes, which is critical for virus entry into host cells for multiplication. It shows high potency with IC50 values in the range of 300-500 nM against multiplication of both influenza pandemic H1N1/2009 and seasonal H3N2/2004 viruses in cell culture. It acts in synergism with either of the two FDA-approved neuraminidase inhibitor (NAI) clinical drugs, zanamivir (Relenza(®)) and oseltamivir carboxylate (active form of Tamiflu(®)), and it has the potential to aid NAI drugs to achieve complete clearance of the virus from the culture. 6SLN-lipo PGA is a new potential candidate drug for influenza control and is an attractive candidate for use in combination with an NAI drug for minimized toxicity, delayed development of resistance, prevention and treatment with the potential for eradication of human influenza. © The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  9. Evidence synthesis and decision modelling to support complex decisions: stockpiling neuraminidase inhibitors for pandemic influenza usage [version 2; referees: 2 approved

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    Samuel I. Watson

    2017-03-01

    Full Text Available Objectives: The stockpiling of neuraminidase inhibitor (NAI antivirals as a defence against pandemic influenza is a significant public health policy decision that must be made despite a lack of conclusive evidence from randomised controlled trials regarding the effectiveness of NAIs on important clinical end points such as mortality. The objective of this study was to determine whether NAIs should be stockpiled for treatment of pandemic influenza on the basis of current evidence. Methods: A decision model for stockpiling was designed. Data on previous pandemic influenza epidemiology was combined with data on the effectiveness of NAIs in reducing mortality obtained from a recent individual participant meta-analysis using observational data. Evidence synthesis techniques and a bias modelling method for observational data were used to incorporate the evidence into the model. The stockpiling decision was modelled for adults (≥16 years old and the United Kingdom was used as an example. The main outcome was the expected net benefits of stockpiling in monetary terms. Health benefits were estimated from deaths averted through stockpiling. Results: After adjusting for biases in the estimated effectiveness of NAIs, the expected net benefit of stockpiling in the baseline analysis was £444 million, assuming a willingness to pay of £20,000/QALY ($31,000/QALY. The decision would therefore be to stockpile NAIs. There was a greater probability that the stockpile would not be utilised than utilised. However, the rare but catastrophic losses from a severe pandemic justified the decision to stockpile. Conclusions: Taking into account the available epidemiological data and evidence of effectiveness of NAIs in reducing mortality, including potential biases, a decision maker should stockpile anti-influenza medication in keeping with the postulated decision rule.

  10. Chimeric Bovine Respiratory Syncytial Virus with Attachment and Fusion Glycoproteins Replaced by Bovine Parainfluenza Virus Type 3 Hemagglutinin-Neuraminidase and Fusion Proteins

    Science.gov (United States)

    Stope, Matthias B.; Karger, Axel; Schmidt, Ulrike; Buchholz, Ursula J.

    2001-01-01

    Chimeric bovine respiratory syncytial viruses (BRSV) expressing glycoproteins of bovine parainfluenza virus type 3 (BPIV-3) instead of BRSV glycoproteins were generated from cDNA. In the BRSV antigenome cDNA, the open reading frames of the major BRSV glycoproteins, attachment protein G and fusion protein F, were replaced individually or together by those of the BPIV-3 hemagglutinin-neuraminidase (HN) and/or fusion (F) glycoproteins. Recombinant virus could not be recovered from cDNA when the BRSV F open reading frame was replaced by the BPIV-3 F open reading frame. However, cDNA recovery of the chimeric virus rBRSV-HNF, with both glycoproteins replaced simultaneously, and of the chimeric virus rBRSV-HN, with the BRSV G protein replaced by BPIV-3 HN, was successful. The replication rates of both chimeras were similar to that of standard rBRSV. Moreover, rBRSV-HNF was neutralized by antibodies specific for BPIV-3, but not by antibodies specific to BRSV, demonstrating that the BRSV glycoproteins can be functionally replaced by BPIV-3 glycoproteins. In contrast, rBRSV-HN was neutralized by BRSV-specific antisera, but not by BPIV-3 specific sera, showing that infection of rBRSV-HN is mediated by BRSV F. Hemadsorption of cells infected with rBRSV-HNF and rBRSV-HN proved that BPIV-3 HN protein expressed by rBRSV is functional. Colocalization of the BPIV-3 glycoproteins with BRSV M protein was demonstrated by confocal laser scan microscopy. Moreover, protein analysis revealed that the BPIV-3 glycoproteins were present in chimeric virions. Taken together, these data indicate that the heterologous glycoproteins were not only expressed but were incorporated into the envelope of recombinant BRSV. Thus, the envelope glycoproteins derived from a member of the Respirovirus genus can together functionally replace their homologs in a Pneumovirus background. PMID:11533200

  11. Supply of neuraminidase inhibitors related to reduced influenza A (H1N1 mortality during the 2009-2010 H1N1 pandemic: an ecological study.

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    Paula E Miller

    Full Text Available BACKGROUND: The influenza A (H1N1 pandemic swept across the globe from April 2009 to August 2010 affecting millions. Many WHO Member States relied on antiviral drugs, specifically neuraminidase inhibitors (NAIs oseltamivir and zanamivir, to treat influenza patients in critical condition. Such drugs have been found to be effective in reducing severity and duration of influenza illness, and likely reduced morbidity during the pandemic. However, it is less clear whether NAIs used during the pandemic reduced H1N1 mortality. METHODS: Country-level data on supply of oseltamivir and zanamivir were used to predict H1N1 mortality (per 100,000 people from July 2009 to August 2010 in forty-two WHO Member States. Poisson regression was used to model the association between NAI supply and H1N1 mortality, with adjustment for economic, demographic, and health-related confounders. RESULTS: After adjustment for potential confounders, each 10% increase in kilograms of oseltamivir, per 100,000 people, was associated with a 1.6% reduction in H1N1 mortality over the pandemic period (relative rate (RR = 0.84 per log increase in oseltamivir supply. While the supply of zanamivir was considerably less than that of oseltamivir in each Member State, each 10% increase in kilogram of active zanamivir, per 100,000, was associated with a 0.3% reduction in H1N1 mortality (RR = 0.97 per log increase. CONCLUSION: While there are limitations to the ecologic nature of these data, this analysis offers evidence of a protective relationship between antiviral drug supply and influenza mortality and supports a role for influenza antiviral use in future pandemics.

  12. Identification of potential B cell epitope determinants by computer techniques, in hemagglutinin-neuraminidase from the porcine rubulavirus La Piedad Michoacan.

    Science.gov (United States)

    Zenteno-Cuevas, Roberto; Huerta-Yepez, Sara; Reyes-Leyva, Julio; Hernández-Jáuregui, Pablo; González-Bonilla, Cesar; Ramírez-Mendoza, Humberto; Agundis, Concepción; Zenteno, Edgar

    2007-01-01

    Hemagglutinin-neuraminidase (HN) from porcine rubulavirus La Piedad Michoacan (RvpLPM) is one of the most antigenic proteins known, and is responsible for virus-host cell interaction. We analyzed the amino acid sequence of HN, using computer-assisted techniques to identify B cell epitopes. From a pool of 18 possible antigenic peptides, we evaluated the antigenicity of the 2 peptides with the highest scores and the 1 with lowest score. Antibodies from RvpLPM-infected pigs recognized the synthesized HN-A, HN-B, and HN-R peptides (optical density [OD]: 0.33 +/- 0.02 for HN-A, 0.20 +/- 0.02 for HN-B, and 0.07 +/- 0.01 for HN-R); bovine serum albumin-coupled HN-A and HN-B induced rabbit anti-RvpLPM antibodies (OD: 0.39 +/- 0.01 for HN-A and 0.35 +/- 0.02 for HN-B). Loop 5 from the outer membrane protein, OmpC, from Salmonella typhi was replaced with HN-B; this protein was then expressed in Escherichia coli UH302. BALB/c mice were challenged intraperitoneally or orogastrically with the fusion protein expressed in E. coli and murine antibodies obtained from both types of administration inhibited virus-hemagglutinating activity, as did the antibodies from RvpLPM-infected swine. These results suggest that HN-A and HN-B are peptides involved in RvpLPM cell carbohydrate recognition, and could therefore be considered potential targets for vaccine and diagnostic procedures development.

  13. Supply of neuraminidase inhibitors related to reduced influenza A (H1N1) mortality during the 2009-2010 H1N1 pandemic: an ecological study.

    Science.gov (United States)

    Miller, Paula E; Rambachan, Aksharananda; Hubbard, Roderick J; Li, Jiabai; Meyer, Alison E; Stephens, Peter; Mounts, Anthony W; Rolfes, Melissa A; Penn, Charles R

    2012-01-01

    The influenza A (H1N1) pandemic swept across the globe from April 2009 to August 2010 affecting millions. Many WHO Member States relied on antiviral drugs, specifically neuraminidase inhibitors (NAIs) oseltamivir and zanamivir, to treat influenza patients in critical condition. Such drugs have been found to be effective in reducing severity and duration of influenza illness, and likely reduced morbidity during the pandemic. However, it is less clear whether NAIs used during the pandemic reduced H1N1 mortality. Country-level data on supply of oseltamivir and zanamivir were used to predict H1N1 mortality (per 100,000 people) from July 2009 to August 2010 in forty-two WHO Member States. Poisson regression was used to model the association between NAI supply and H1N1 mortality, with adjustment for economic, demographic, and health-related confounders. After adjustment for potential confounders, each 10% increase in kilograms of oseltamivir, per 100,000 people, was associated with a 1.6% reduction in H1N1 mortality over the pandemic period (relative rate (RR) = 0.84 per log increase in oseltamivir supply). While the supply of zanamivir was considerably less than that of oseltamivir in each Member State, each 10% increase in kilogram of active zanamivir, per 100,000, was associated with a 0.3% reduction in H1N1 mortality (RR = 0.97 per log increase). While there are limitations to the ecologic nature of these data, this analysis offers evidence of a protective relationship between antiviral drug supply and influenza mortality and supports a role for influenza antiviral use in future pandemics.

  14. Análise de polimorfismos do gene que codifica a proteína B do surfactante: comparação entre recém-nascidos pré-termo com e sem síndrome do desconforto respiratório

    OpenAIRE

    Priscila Pinheiro Ribeiro Lyra

    2010-01-01

    A síndrome do desconforto respiratório (SDR) é causada pela deficiência transitória de surfactante pulmonar em recém-nascidos (RN) prematuros nos primeiros dias de vida. Estudos sugerem que a etiologia da SDR seja multifatorial e multigênica. A proteína B do surfactante (SP-B) é fundamental para o metabolismo do surfactante e para uma função pulmonar normal. A presença de polimorfismos e mutações em genes dos componentes do surfactante, particularmente no gene da SP-B, parece estar associada ...

  15. Regulation of Na+ fluxes in plants

    Directory of Open Access Journals (Sweden)

    Frans eMaathuis

    2014-09-01

    Full Text Available When exposed to salt, every plant takes up Na+ from the environment. Once in the symplast, Na+ is distributed within cells and between different tissues and organs. There it can help to lower the cellular water potential but also exert potentially toxic effects. Control of Na+ fluxes is therefore crucial and indeed, research shows that the divergence between salt tolerant and salt sensitive plants is not due to a variation in transporter types but rather originates in the control of uptake and internal Na+ fluxes. A number of regulatory mechanisms has been identified based on signalling of Ca2+, cyclic nucleotides, reactive oxygen species, hormones, or on transcriptional and post translational changes of gene and protein expression. This review will give an overview of intra- and intercellular movement of Na+ in plants and will summarise our current ideas of how these fluxes are controlled and regulated in the early stages of salt stress.

  16. Análise de polimorfismos do gene da kapa-caseína em fêmeas da raça Nelore e efeito sobre o peso à desmama de suas progênies Polimorphism analysis of kappa -casein gene on Nelore females and effect on weaning weight of the calves

    Directory of Open Access Journals (Sweden)

    F.J.C. Faria

    1999-08-01

    Full Text Available Informações sobre peso à desmama de um rebanho Nelore, após ajuste para idade padrão de 205 dias, sexo da cria, idade da mãe, touro e mês de desmama, foram utilizadas para separar as reprodutrizes em dois grupos, cujos produtos diferiam em peso. As médias ajustadas pelo método dos quadrados mínimos e erros-padrão para os grupos pesado (P e leve (L foram 163,21±2,18kg e 134,44±2,18kg, respectivamente, com 41 animais em cada grupo. Colheram-se amostras de sangue das reprodutrizes para o estudo de polimorfismos do gene da kapa-caseína, pela técnica de polymerase chain reaction-restriction fragment length polymorphism. A análise de um segmento do éxon IV do gene da kapa-caseína identificou os genótipos 71AA, 10AB e 1BB, com as freqüências de 0,92 e 0,08 para os alelos A e B, respectivamente. Os pesos à desmama dos produtos dos genótipos AA e AB foram, respectivamente, 147,73±2,38kg e 156,76±6,35kg. Os grupos P e L não diferiram entre si quanto às freqüências alélicas apresentadas para esse gene. O genótipo das reprodutrizes não afetou o peso à desmama de suas crias, existindo, portanto, outros efeitos genéticos e não genéticos de maior magnitude.Data from a Nelore herd were used after adjustment of the weaning weight for 205 days of age, sex, age of dam, sire and month of weaning resulting two groups of cows that differed in weaning weight of their calves. The least square means (LSM and standard error (SE were 163.21±2.18kg and 134.44±2.18kg, for heavy (H and light (L groups, with 41 animals each one. These animals were genotyped for DNA polymorphisms of kappa-casein gene, using PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism. The segment analysis of kappa-casein IV exon identified the genotypes 71AA, 10AB and 1BB, with frequencies of 0.92 and 0.08 for A and B alleles, respectively. The LSM±SE for weaning weight were 147.73±2.38kg and 156.76± 6.35kg for calves of AA and AB

  17. Análise do perfil de expressão dos genes da cana-de-açúcar envolvidos na interação com Leifsonia xyli subsp: xyli Differential gene expression in sugar cane infected by Leifsonia xyli subsp: xyli

    Directory of Open Access Journals (Sweden)

    Maria Inês Tiraboschi Ferro

    2007-06-01

    Full Text Available Utilizando a técnica de macroarranjos de cDNA em membranas de náilon, analisou-se o perfil de expressão de 3.575 ESTs ("Expressed Sequence Tags" de cana-de-açúcar, oriundas do projeto SUCEST, em duas variedades, uma tolerante (SP80-0185 e outra suscetível (SP70-3370 ao Raquitismo da Soqueira. Foram analisadas amostras foliares de plantas inoculadas com Leifsonia xyli subsp. xyli., agente etiológico do Raquitismo, contrastadas com plantas não inoculadas (controle, para cada variedade, marcadas com sondas de cDNA e hibridizadas contra os macroarranjos. Após as hibridizações e análises estatísticas dos dados foi possível identificar 49 ESTs com expressão alterada, sendo 44 na variedade tolerante (41 ESTs induzidos e 3 reprimidos e 5 na variedade suscetível (2 ESTs induzidos e 3 reprimidos. Os resultados obtidos sugerem que a tolerância da variedade SP80-0185 de cana-de-açúcar à bactéria fitopatogênica pode estar relacionada com a percepção de sinais extracelulares, visto que ESTs relacionados a vias de transdução de sinais apresentaram expressão gênica induzida na variedade tolerante, os quais codificam para uma EST com similaridade à H+-ATPase da membrana plasmática, fatores de transcrição G-box, OsNAC6, "DNA binding", família MYB e "Zinc Finger" e ainda uma EST com similaridade ao fator de ligação ao G-Box, o qual corresponde a uma seqüência de DNA cis presente em vários promotores de plantas e requerido para o reconhecimento de muitos estímulos ambientais. Na variedade suscetível foi reprimido uma EST com similaridade à lipase. Esta enzima, também de membrana, faz parte da síntese do jasmonato, o qual ativa as defesas vegetais contra patógenos de plantas. Possíveis funções para os genes induzidos ou reprimidos nas cultivares de cana tolerante ou resistente ao Raquitismo são discutidas neste trabalho.The macroarray nylon membrane technology was used to study the differential gene expression of 3

  18. Genes and Gene Therapy

    Science.gov (United States)

    ... correctly, a child can have a genetic disorder. Gene therapy is an experimental technique that uses genes to ... or prevent disease. The most common form of gene therapy involves inserting a normal gene to replace an ...

  19. Glycosylation alters steady-state inactivation of sodium channel Nav1.9/NaN in dorsal root ganglion neurons and is developmentally regulated.

    Science.gov (United States)

    Tyrrell, L; Renganathan, M; Dib-Hajj, S D; Waxman, S G

    2001-12-15

    Na channel NaN (Na(v)1.9) produces a persistent TTX-resistant (TTX-R) current in small-diameter neurons of dorsal root ganglia (DRG) and trigeminal ganglia. Na(v)1.9-specific antibodies react in immunoblot assays with a 210 kDa protein from the membrane fractions of adult DRG and trigeminal ganglia. The size of the immunoreactive protein is in close agreement with the predicted Na(v)1.9 theoretical molecular weight of 201 kDa, suggesting limited glycosylation of this channel in adult tissues. Neonatal rat DRG membrane fractions, however, contain an additional higher molecular weight immunoreactive protein. Reverse transcription-PCR analysis did not show additional longer transcripts that could encode the larger protein. Enzymatic deglycosylation of the membrane preparations converted both immunoreactive proteins into a single faster migrating band, consistent with two states of glycosylation of Na(v)1.9. The developmental change in the glycosylation state of Na(v)1.9 is paralleled by a developmental change in the gating of the persistent TTX-R Na(+) current attributable to Na(v)1.9 in native DRG neurons. Whole-cell patch-clamp analysis demonstrates that the midpoint of steady-state inactivation is shifted 7 mV in a hyperpolarized direction in neonatal (postnatal days 0-3) compared with adult DRG neurons, although there is no significant difference in activation. Pretreatment of neonatal DRG neurons with neuraminidase causes an 8 mV depolarizing shift in the midpoint of steady-state inactivation of Na(v)1.9, making it indistinguishable from that of adult DRG neurons. Our data show that extensive glycosylation of rat Na(v)1.9 is developmentally regulated and changes a critical property of this channel in native neurons.

  20. Genetic grouping of avian infectious bronchitis virus isolated in Brazil based on RT-PCR/RFLP analysis of the S1 gene Agrupamento genético de isolados do vírus da bronquite infecciosa das aves no Brasil com base na análise do gene S1 por RT-PCR-RFLP

    Directory of Open Access Journals (Sweden)

    Maria de Fátima S. Montassier

    2008-03-01

    Full Text Available Twelve Brazilian isolates and one reference vaccine strain of avian infectious bronchitis virus (IBV were propagated in embryonating chicken eggs. The entire S1 glycoprotein gene of these viruses was analysed by reverse-transcriptase-polymerase chain reaction and restriction fragment length polymorphism (RT-PCR-RFLP, using the restriction enzymes HaeIII, XcmI and BstyI. The RFLP patterns led to the classification of these isolates into five distinct genotypes: A, B, C, D and Massachusetts. Five of twelve isolates were grouped in Massachusetts genotype and the remaining seven viruses were classified into four distinct genotypes: A (2, B (2, C (2 or D (1. Such genotyping classification agreed with previous immunological analysis for most of these viruses, highlighting the occurrence of a relevant variability among the IBV strains that are circulating in Brazilian commercial poultry flocks.Doze isolados de campo do Brasil e uma estirpe de referência vacinal do vírus da bronquite infecciosa das aves (VBI foram propagadas em ovos embrionados SPF. O gene S1 dessas amostras foi analisado por RT-PCR seguido de RFLP, empregando-se as enzimas de restrição HaeIII, XcmI e BstyI. Observou-se a existência de cinco genotipos diferentes: M (Massachusetts, A , B, C e D. Cinco dos doze isolados de campo do VBI foram classificados no genótipo Massachusetts e os sete vírus restantes foram classificados em quatro genotipos diferentes; A (2, B (2, C (2 ou D (1. Os resultados desta genotipagem concordam com os dados obtidos na análise imunológica previamente realizada para a maior parte destes vírus, destacando a ocorrência de uma variabilidade marcante entre os isolados do VBI que estão circulando nas granjas avícolas comerciais do Brasil.

  1. Actividad de la proteína transportadora de ésteres de colesterol. Polimorfismos del gen en pacientes colombianos con enfermedad coronaria Activity of cholesteryl ester transfer protein. Gene polymorphism in colombian patients with coronary artery disease

    Directory of Open Access Journals (Sweden)

    Alejandra M Giraldo

    2012-08-01

    Full Text Available Introducción: la literatura relaciona la actividad de la proteína transportadora de ésteres de colesterol (CETP con enfermedad coronaria, por reducir el colesterol en las lipoproteínas de alta densidad. Adicionalmente, estudios recientes han identificado variaciones en el gen de la CETP, aunque el papel funcional de algunas de estas variantes sobre la actividad enzimática y la enfermedad coronaria, es desconocido. Objetivos: examinar la asociación de los polimorfismos TaqIB, MspI y RsaI del gen de la CETP y la actividad de la enzima con enfermedad coronaria. Métodos: se evaluó la asociación entre la actividad de la enzima y los polimorfismos TaqIB, MspI y RsaI, en pacientes con obstrucción coronaria documentada por angiografía. Resultados: la angiografía permitió clasificar a los pacientes en dos grupos: uno (213 individuos con obstrucción coronaria no significativa (OC 50%. La edad fue significantemente mayor en el último grupo en comparación con el primero. La actividad de la CETP fue 95,8 y 94,7 pmol/μL.h, para los grupos OC 50%, respectivamente. Solo se encontró diferencia significativa entre los alelos M1 y M2 en la población general. Conclusión: no se halló asociación entre la actividad de la CETP, los polimorfismos TaqBI, MspI, Rsal y la obstrucción coronaria. En este trabajo se describen por primera vez los niveles de CETP en los polimorfismos TaqIB, MspI, Rsal para un grupo de pacientes colombianos. Se debe refinar la descripción del evento coronario, el contexto metabólico de los pacientes y el estudio de haplotipos para encontrar relaciones con enfermedad coronaria.Introduction: literature links the activity of cholesteryl ester transfer protein (CETP with coronary heart disease by lowering cholesterol in high density lipoproteins. Additionally, recent studies have identified variations in the CETP gene, although the functional role of some of these variants on enzyme activity and coronary heart disease is

  2. Isolation of Arachis hypogaea Na /H antiporter and its expression ...

    African Journals Online (AJOL)

    Jane

    2011-10-24

    Oct 24, 2011 ... China. Accepted 23 September, 2011. The plant Na+/H+ antiporter gene plays a major role in ... plants have developed various adaptation mechanisms ..... of transgenic maize with improved salt tolerance by the introduction.

  3. Tratamientos farmacológicos en la coadministración de alcohol y cocaína: efectos en la expresión de genes en el córtex prefrontal de la rata

    OpenAIRE

    Echeverry Alzate, Víctor

    2016-01-01

    El consumo de sustancias psicoactivas, y en concreto el consumo simultáneo de alcohol y cocaína, es una práctica frecuente en la población general. Así mismo, se ha visto que el consumo de cocaína incrementa el riesgo de desarrollar dependencia de alcohol, y viceversa. Actualmente existen 4 tratamientos farmacológicos aprobados para la dependencia de alcohol, siendo el disulfiram el primer tratamiento aprobado, posteriormente lo fueron la naltrexona y el acamprosato, y recientemente el nalmef...

  4. Interação entre qualidade do meio ambiente, estresse e a variação do gene APOE na determinação da suscetibilidade à fibromialgia Association between environmental quality, stress and APOE gene variation in fibromyalgia susceptibility determination

    Directory of Open Access Journals (Sweden)

    Roze Mary Ribas Becker

    2010-12-01

    Full Text Available INTRODUÇÃO: A fibromialgia se trata de uma desordem multifatorial, cuja etiologia reside na interação entre a susceptibilidade genética e o ambiente. No entanto, poucos trabalhos procuram detectar quais seriam os fatores considerados de risco. OBJETIVO: Investigar a influência genética e sua interação com qualidade ambiental e com estresse como possíveis fatores de risco para o desenvolvimento da fibromialgia. PACIENTES E MÉTODOS: Neste estudo transversal, foram investigados dois grupos de mulheres, sendo 47 com diagnóstico clínico de fibromialgia, e 41 mulheres do grupo controle, todas da comunidade de Novo Hamburgo, RS. O polimorfismo do gene da apolipoproteína E (APOE foi analisado, a partir do DNA extraído do sangue total de ambas as amostras. Os fatores ambientais foram avaliados através do inventário de sintomas para adultos de Lipp (ISSL, para a averiguação do estresse comportamental, e da aplicação do domínio V do WHOQOL-100. RESULTADOS: Dentre as pacientes, foram encontradas mais mulheres com níveis altos de estresse, quando comparado à amostra controle (P INTRODUCTION: Fibromyalgia is a multifactorial disease, of which etiology is based on interaction between genetic susceptibility and environment. However, few studies attempted to identify the risk factors. OBJECTIVE: To investigate the genetic influence and its interaction with environmental quality and stress, as possible risk factors for fibromyalgia development. PATIENTS AND METHODS: This cross-sectional study investigated two groups of women, of which 47 had a clinical diagnosis of fibromyalgia, and 41 women comprising thre control group, all from the town of Novo Hamburgo, RS. The apolipoprotein E (APOE gene polymorphism was analyzed in DNA extracted from total blood, in both samples. Environmental factors were studied through Lipp's Inventory of Stress Symptoms for Adults and by applying the WHOQOL-100 domain V. RESULTS: Among the patients, more women

  5. Association of the heart fatty acid-binding protein gene with quality of carcass and meat traits in pigs Associação entre o gene da proteína de ligação de ácidos graxos - coração com características de carcaça e qualidade da carne em suínos

    Directory of Open Access Journals (Sweden)

    F.C. Figueiredo

    2008-04-01

    Full Text Available The heart fatty acid-binding protein (HFABP gene was sequenced in parental animals of a F2 crossing of boars of the Brazilian native Piau breed with commercial sows (Landrace x Large White Pietrain. Primers used for PCR were designed to amplify four exons of the gene. The PCR products were sequenced and compared with the GenBank sequences. Differences between the generated sequences and the GenBank sequences were observed for both genetic groups. A total of 246 F2 animals were genotyped using the Hinf I restriction enzyme. Two genotypes were identified, 198 being animals HH and 48 Hh. The Hinf I SNP was significantly associated with weights of loin (bone-in (PO gene da proteína de ligação de ácidos graxos - coração foi seqüenciado em animais parentais de um cruzamento F2 entre varrões da raça nativa brasileira Piau e fêmeas comerciais (Landrace x Large White x Pietrain. Os primers utilizados na reação em cadeia da polimerase foram desenhados para amplificarem os quatro éxons do gene. Os fragmentos amplificados foram seqüenciados e comparados com a seqüência do gene depositada no GenBank. Foram observadas divergências entre as seqüências geradas e as do GenBank para ambos os grupos genéticos. Foram genotipados 246 animais F2 utilizando-se a enzima Hinf I. Dois genótipos foram identificados, sendo 198 animais HH e 48 animais Hh. O polimorfismo apresentou efeito sobre o peso total do carré (P<0,05, o peso da papada (P<0,05, o peso do filezinho (P<0,10 e o peso dos rins (P<0,01. Os resultados indicam que o gene da H-FABP apresenta potencial para aplicação em programas de seleção assistida por marcadores moleculares em suínos.

  6. Testing Na+ in blood

    OpenAIRE

    Lava, Sebastiano A.G.; Bianchetti, Mario G.; Milani, Gregorio P.

    2016-01-01

    Abstract Both direct potentiometry and indirect potentiometry are currently used for Na+ testing in blood. These measurement techniques show good agreement as long as protein and lipid concentrations in blood remain normal. In severely ill patients, indirect potentiometry commonly leads to relevant errors in Na+ estimation: 25% of specimens show a disagreement between direct and indirect potentiometry, which is ?4 mmol/L (mostly spuriously elevated Na+ level due to low circulating albumin con...

  7. Supply of neuraminidase inhibitors related to reduced influenza A (H1N1) mortality during the 2009-2010 H1N1 pandemic: summary of an ecological study.

    Science.gov (United States)

    Miller, Paula E; Rambachan, Aksharananda; Hubbard, Roderick J; Li, Jiabai; Meyer, Alison E; Stephens, Peter; Mounts, Anthony W; Rolfes, Melissa A; Penn, Charles R

    2013-09-01

    When the influenza A (H1N1) pandemic spread across the globe from April 2009 to August 2010, many WHO Member States used antiviral drugs, specifically neuraminidase inhibitors (NAIs) oseltamivir and zanamivir, to treat influenza patients in critical condition. Antivirals have been found to be effective in reducing severity and duration of influenza illness, and likely reduce morbidity; however, it is unclear whether NAIs used during the pandemic reduced H1N1 mortality. To assess the association between antivirals and influenza mortality, at an ecologic level, country-level data on supply of oseltamivir and zanamivir were compared to laboratory-confirmed H1N1 deaths (per 100 000 people) from July 2009 to August 2010 in 42 WHO Member States. From this analysis, it was found that each 10% increase in kilograms of oseltamivir, per 100 000 people, was associated with a 1·6% reduction in H1N1 mortality over the pandemic period [relative rate (RR) = 0·84 per log increase in oseltamivir supply]. Each 10% increase in kilogram of active zanamivir, per 100 000, was associated with a 0·3% reduction in H1N1 mortality (RR = 0·97 per log increase). While limitations exist in the inference that can be drawn from an ecologic evaluation, this analysis offers evidence of a protective relationship between antiviral drug supply and influenza mortality and supports a role for influenza antiviral use in future pandemics. This article summarises the original study described previously, which can be accessed through the following citation: Miller PE, Rambachan A, Hubbard RJ, Li J, Meyer AE, et al. (2012) Supply of Neuraminidase Inhibitors Related to Reduced Influenza A (H1N1) Mortality during the 2009-2010 H1N1 Pandemic: An Ecological Study. PLoS ONE 7(9): e43491. © 2013 Blackwell Publishing Ltd.

  8. VPLIV AMBROZIJE NA ZDRAVJE

    OpenAIRE

    Tramšek, Dejan

    2015-01-01

    Izhodišča: V raziskavi smo se osredotočili na opis različnih vrst ambrozije. Opisali smo tudi rastline, ki so v veliki meri podobne ambroziji. Ob vsem tem pa seveda vpliv ambrozije na zdravje ljudi, predvsem alergij. Predstavili smo posamezne alergije, opisali način preprečevanja in zdravljenja le-teh in simptome, ki alergije spremljajo. Za omejitev in zmanjšanje pojava alergij na ambrozijo pa je to rastlino potrebno odstraniti oz. zamejiti. Odločili smo se, da v diplomskem delu predstavimo ...

  9. Mumps virus F gene and HN gene sequencing as a molecular tool to study mumps virus transmission.

    Science.gov (United States)

    Gouma, Sigrid; Cremer, Jeroen; Parkkali, Saara; Veldhuijzen, Irene; van Binnendijk, Rob S; Koopmans, Marion P G

    2016-11-01

    Various mumps outbreaks have occurred in the Netherlands since 2004, particularly among persons who had received 2 doses of measles, mumps, and rubella (MMR) vaccination. Genomic typing of pathogens can be used to track outbreaks, but the established genotyping of mumps virus based on the small hydrophobic (SH) gene sequences did not provide sufficient resolution. Therefore, we expanded the sequencing to include fusion (F) gene and haemagglutinin-neuraminidase (HN) gene sequences in addition to the SH gene sequences from 109 mumps virus genotype G strains obtained between 2004 and mid 2015 in the Netherlands. When the molecular information from these 3 genes was combined, we were able to identify separate mumps virus clusters and track mumps virus transmission. The analyses suggested that multiple mumps virus introductions occurred in the Netherlands between 2004 and 2015 resulting in several mumps outbreaks throughout this period, whereas during some local outbreaks the molecular data pointed towards endemic circulation. Combined analysis of epidemiological data and sequence data collected in 2015 showed good support for the phylogenetic clustering. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Expressão dos genes nod de Rhizobium tropici, R. etli e R. leguminosarum bv. phaseoli e estabelecimento da nodulação do feijoeiro na presença de exsudatos de sementes de Mimosa flocculosa e Leucaena leucocephala

    Directory of Open Access Journals (Sweden)

    F. M. Mercante

    2000-06-01

    Full Text Available Na etapa inicial da troca de sinais moleculares entre macro e microssimbiontes, a interação do feijoeiro e estirpes de Rhizobium tropici, R. etli e R. leguminosarum bv. phaseoli foi avaliada pela expressão dos genes nod de estirpes bacterianas, contendo a fusão nodA::gusA. Esta avaliação foi efetuada por meio da atividade da enzima ß-glucuronidase, utilizando, como indutores, exsudatos liberados pelas sementes de Mimosa flocculosa e Leucaena leucocephala. Além disso, avaliou-se o efeito da adição desses exsudatos no estabelecimento da nodulação do feijoeiro, cv. Carioca. Nos testes "in vitro", a mistura de exsudatos de sementes de feijoeiro e M. flocculosa promoveu aumentos sinergísticos significativos na expressão dos genes nod, tanto das estirpes de R. tropici (CIAT 899/pGUS 32 e F 98.5/pGUS 32 quanto de R. etli (CFN 42/pGUS 32. Em condições controladas, a adição dos exsudatos, tanto de M. flocculosa quanto de L. leucocephala, proporcionou aumento significativo na nodulação inicial do feijoeiro, quando foi inoculada a estirpe CFN 42 (R. etli. A nodulação do feijoeiro cultivado em vasos com solo não foi inibida pelo suprimento de N-mineral, quando se inoculou a estirpe CIAT 899 (R. tropici e foram fornecidos exsudatos de sementes de M. flocculosa.

  11. Perfil transcricional dos genes envolvidos na via de biossíntese de carotenóides e quantificação dos metabólitos em laranjas de polpa vermelha

    OpenAIRE

    Deborah Sanae Nishimura

    2012-01-01

    A combinação do aumento da população mundial e a melhoria nos padrões de vida estão levando a uma maior demanda por alimentos com alto valor nutricional. As variedades de laranjas de polpa vermelha são as únicas laranjas que acumulam licopeno na polpa e no suco. O licopeno é um carotenóide considerado como um composto antioxidante, com ação potencial na prevenção de alguns tipos de câncer. Apesar do valor nutricional agregado as laranjas de polpa vermelha, a caracterização fisiológica e mo...

  12. O papel do gene e da molécula HLA-G na expressão clínica das doenças reumatológicas

    OpenAIRE

    Brenol,Claiton Viegas; Veit,Tiago Degani; Chies,José Artur Bogo; Xavier,Ricardo Machado

    2012-01-01

    O antígeno leucocitário humano G (HLA-G) é uma molécula não clássica de complexo principal de histocompatibilidade (MHC) de classe I, caracterizada por baixo polimorfismo em sua região codificadora, um padrão de distribuição tecidual limitado em condições fisiológicas e expressão por meio de isoformas solúveis e acopladas à superfície de membranas por meio de splicing alternativo. O HLA-G é bastante conhecido por estar envolvido na indução e na manutenção da tolerância entre o sistema imunoló...

  13. Međudjelovanje genskog polimorfizma filagrina i načina života kod atopijskih bolesti u mladoj odrasloj populaciji [Interaction of filaggrin gene polymorphisms and environmental factors with atopic diseases in young adult population

    OpenAIRE

    Sabolić Pipinić, Ivana

    2015-01-01

    Pathogenesis of atopic disorders such as atopic rhinitis, atopic asthma and atopic dermatitis lies in gene-environment interactions. Hypothesis of this study is that in Croatian population filaggrin gene (FLG) polymorphisms (R501X, 2282del4, R2447X i S3247X) contribue to atopic diseases, and that this contribution is modulated by environmetal factors (settlement type, heating type, animal contact, smoking, physical activity, BMI, nutrition). The study enrolled 439 freshmen students at the Uni...

  14. Functional coupling of renal K+ and Na+ handling causes high blood pressure in Na+ replete mice.

    Science.gov (United States)

    Vitzthum, Helga; Seniuk, Anika; Schulte, Laura Helene; Müller, Maxie Luise; Hetz, Hannah; Ehmke, Heimo

    2014-03-01

    A network of kinases, including WNKs, SPAK and Sgk1, is critical for the independent regulation of K+ and Na+ transport in the distal nephron. Angiotensin II is thought to act as a key hormone in orchestrating these kinases to switch from K+ secretion during hyperkalaemia to Na+ reabsorption during intravascular volume depletion, thus keeping disturbances in electrolyte and blood pressure homeostasis at a minimum. It remains unclear, however, how K+ and Na+ transport are regulated during a high Na+ intake, which is associated with suppressed angiotensin II levels and a high distal tubular Na+ load. We therefore investigated the integrated blood pressure, renal, hormonal and gene and protein expression responses to large changes of K+ intake in Na+ replete mice. Both low and high K+ intake increased blood pressure and caused Na+ retention. Low K+ intake was accompanied by an upregulation of the sodium-chloride cotransporter (NCC) and its activating kinase SPAK, and inhibition of NCC normalized blood pressure. Renal responses were unaffected by angiotensin AT1 receptor antagonism, indicating that low K+ intake activates the distal nephron by an angiotensin-independent mode of action. High K+ intake was associated with elevated plasma aldosterone concentrations and an upregulation of the epithelial sodium channel (ENaC) and its activating kinase Sgk1. Surprisingly, high K+ intake increased blood pressure even during ENaC or mineralocorticoid receptor antagonism, suggesting the contribution of aldosterone-independent mechanisms. These findings show that in a Na+ replete state, changes in K+ intake induce specific molecular and functional adaptations in the distal nephron that cause a functional coupling of renal K+ and Na+ handling, resulting in Na+ retention and high blood pressure when K+ intake is either restricted or excessively increased.

  15. Genes associados à virulência e multirresistência de antimicrobianos em linhagens Trueperella Pyogenes isoladas de mastite e outras afecções em animais domésticos

    OpenAIRE

    RISSETI, Rafaela Mastrangelo

    2015-01-01

    Trueperella pyogenes are opportunistic bacterium characterized by suppurative infections in domestic animals, commonly refractory to conventional therapy. Recently, genes which encode exotoxin pyolysin (plo), and factors that promote adhesion of T. pyogenes to host cells, such as fimbriae (fimA, fimC, fimE, fimG), neuraminidases (nanH, nanP), and collagen-binding protein (cbpA) have been associated to virulence of pathogen. The aim of present study was investigate occurrence of multi-drug res...

  16. Efeito dos ácidos graxos n-3 e n-6 na expressão de genes do metabolismo de lipídeos e risco de aterosclerose Effects of n-3 and n-6 fatty acids on the expression of genes involved in the lipid metabolism and risk of atherosclerosis

    Directory of Open Access Journals (Sweden)

    Helena Fonseca Raposo

    2010-10-01

    Full Text Available A aterosclerose, principal responsável pela patogênese do infarto miocárdico e cerebral, bem como pela gangrena e por outras doenças vasculares periféricas, permanece como principal causa de morbidade e mortalidade nas populações "ocidentalizadas". Estima-se que 17,5 milhões de pessoas morreram por doenças cardiovasculares em 2005, o que representou 30% das causas de morte nesse ano, e que, em 2015, 20 milhões de pessoas morrerão por doenças cardiovasculares no mundo. Os ácidos graxos n-3, principalmente os de cadeia longa, encontrados nos peixes, têm-se mostrado particularmente úteis na prevenção e tratamento de doenças como dislipidemias, diabetes mellitus e obesidade, apresentando importante efeito cardioprotetor. Nesse contexto, pesquisas têm evidenciado que ao menos parte dos benefícios dos ácidos graxos eicosapentaenóico e docosahexaenóico sobre o risco de doenças cardiovasculares é decorrente da modulação de genes responsivos aos receptores ativados por proliferadores de peroxissomos e envolvidos no metabolismo lipídico. Nesta revisão, pretende-se expor alguns mecanismos de ação dos ácidos graxos n-3 e n-6 sobre o metabolismo de lipídeos e de lipoproteínas. Conclui-se que muitos aspectos que contribuem para o risco de doenças cardiovasculares são afetados pela ingestão de n-3. Além da redução de triglicérides, fatores como o aumento de adiponectina, a redução da concentração de colesterol plasmático e a melhora do transporte reverso de colesterol também são responsáveis pela redução do risco de aterosclerose promovida pelos ácidos graxos n-3. No entanto, ainda são necessários estudos adicionais para definir mais claramente os mecanismos celulares e moleculares responsáveis pelo efeito cardioprotetor dos ácidos graxos n-3.Atherosclerosis, the main cause of myocardial infarction, stroke, gangrene and other peripheral vascular diseases, also persists as the main cause of morbidity and

  17. At NA2

    CERN Multimedia

    1977-01-01

    One of the NA2 calorimeter sections is moved in. The NA2 calorimeter was divided in two halves, to the left and the right of the beam, each half consisting of sheets of passive high Z material interleaved with blades of plastic scintillators. The photo shows on the right, the upstream 'electron' module with eleven lead plates for a total radiation length of 20.

  18. Modulation of oxidative and glycolytic skeletal muscle fibers Na+/H+ exchanger1 (NHE1) and Na+/HCO3- co-transporter1 (NBC1) genes and proteins expression in type 2 diabetic rat (Streptozotocin + high fat diet) following long term endurance training.

    Science.gov (United States)

    Monazzami, A; Rajabi, H; Ghrakhanlou, R; Yari, K; Rahimi, Z

    2017-05-20

    Diabetes is known to alter both oxidative and glycolytic pathways in a fiber type-dependent manner. The aim of present study was to investigate the effects of endurance training on muscle NHE1 and NBC1 genes and proteins expression in type 2 diabetic rats. Male wistar rats (n=30), 4 weeks old and 95.7±10.8g, were randomly selected and divided into control, diabetic without training and diabetic with training groups. Diabetes was induced by injection of low dose of streptotozin and feeding with high-fat diet. The Endurance training was performed for 7 weeks that started with relatively low speed and duration of 20 m min-1 for 20 min in the first week and gradually reached to 30 m min-1 for 35min in the last week. NHE1 and NBC1 genes and proteins expression were determined by Real time-PCR and western blotting techniques, respectively, in Soleus as an oxidative and EDL (Extensor digitorum longus) as a glycolytic muscle preparation. NHE1 mRNA and protein expression reduced significantly in EDL and Soleus in the diabetic without training group compared with the control group. However, reduction in the expression of NBC1 gene and protein in the diabetic without training group compared to controls did not significant. Endurance training increased NHE1 and NBC1 genes and proteins expression in both EDL and Soleus in the diabetic training group compared to control groups. In conclusion, endurance training may improve the capacity of pHi regulation in muscles by lactate-independent pathway.

  19. Gêneros orais na escola

    OpenAIRE

    Teixeira,Lucia

    2012-01-01

    O artigo analisa a utilização da noção de gênero na escola fundamental, detendo-se sobre o ensino dos gêneros orais. Apresenta postulados teóricos de Bakhtin e incorpora a contribuição da semiótica discursiva e as sugestões metodológicas da pedagogia de ensino de línguas desenvolvida na Universidade de Genebra. Em seguida, propõe exemplo prático de aplicação do conceito ao desenvolvimento da expressão oral em sala de aula.

  20. Detecção dos genes codificantes da toxina CDT, e pesquisa de fatores que influenciam na produção de hemolisinas em amostras de Campylobacter jejuni de origem avícola

    OpenAIRE

    Michele M. Trindade; Gustavo Perdoncini; Sierra-Arguello, Yuli M.; Maristela Lovato; Anderlise Borsoi; Nascimento,Vladimir P.

    2015-01-01

    Resumo: Membros termofílicos do gênero Campylobacter são reconhecidos como importantes enteropatógenos para o ser humano e animais. A grande diversidade ecológica destes micro-organismos em diferentes habitats tais como água, animais e alimentos predispõem ao aparecimento de novos fatores de virulência. Este trabalho teve por objetivo detectar os genes codificantes da Toxina Distensiva Citoletal (CDT) por meio da técnica de PCR, pesquisar a atividade de hemolisinas e a influência de soluções ...

  1. Gene Therapy

    Science.gov (United States)

    Gene therapy Overview Gene therapy involves altering the genes inside your body's cells in an effort to treat or stop disease. Genes contain your ... that don't work properly can cause disease. Gene therapy replaces a faulty gene or adds a new ...

  2. At NA5

    CERN Multimedia

    1978-01-01

    The NA5 experiment is shown at the start of the setting up in beam H2 of hall EHN1. It was designed by the Bari- Cracow-Liverpool-Munich (MPI) Collaboration to study hard hadron-hadron collisions with a streamer chamber vertex spectrometer and a calorimeter trigger. The beam from target T2 enters from the left. E

  3. Riso na epilepsia

    Directory of Open Access Journals (Sweden)

    Edymar Jardim

    1967-06-01

    Full Text Available São estudados três casos de síndrome convulsiva temporal, com manifestações concomitantes de riso na sua fase inicial. As características principais foram a imotivação e á incoercibilidade do riso. Esses sintomas desapareceram com o uso de anticonvulsivantes.

  4. Biotecnologia na agricultura

    Directory of Open Access Journals (Sweden)

    Helaine Carrer

    2010-01-01

    Full Text Available A expectativa de o crescimento populacional atingir 9 bilhões de habitantes em 2050 em adição às questões da sustentabilidade e do aquecimento global nos desafiam a aumentar a oferta de alimentos. Uma metodologia alternativa que contribua para a redução do impacto desse cenário envolve a biotecnologia, que, nas últimas décadas, trouxe marcantes oportunidades tecnológicas na agricultura, resultando em relevante desenvolvimento na obtenção de novas variedades de plantas, na melhoria da qualidade de diversos alimentos e atualmente também na bioenergia. As técnicas biotecnológicas envolvendo os marcadores moleculares, a genômica e a transformação genética estão transformando a agricultura e são discutidas neste artigo.The expected population growth to reach 9 billion by 2050 in addition to issues of sustainability and global warming challenges us to increase the supply of food. An alternative approach to help reducing the impact of this scenario involves biotechnology which in recent decades has brought remarkable technological opportunities in the agriculture that resulted in relevant development in obtaining new plant varieties, improved quality of different foods, and now also in bioenergy. The biotechnology techniques involving molecular markers, genomics and genetic transformation are transforming agriculture and will be discussed in this article.

  5. At NA3

    CERN Multimedia

    1977-01-01

    The NA3 experiment was setup in beam H8 in the Hall EHN1 by the CERN-College de France-Orsay-Palaiseau-Saclay Collaboration to study the hadronic production of high pT leptons and hadrons. The photo shows in the background (centre) the electromagnetic calorimeter.

  6. Conectando famílias de construções genéticas: testes de DNA na reunificação da família somali na Finlândia Connecting genes-building families: DNA testing in somali family reunification in Finland

    Directory of Open Access Journals (Sweden)

    Petri Hautaniemi

    2007-12-01

    Full Text Available Os temas centrais desse artigo, reunificação familiar em geral e teste de DNA em particular, surgiram a partir de uma pesquisa em andamento acerca de jovens da Somália na Finlândia. Desde 1996, realizo uma pesquisa etnográfica - em escolas, clubes de jovens, ruas e cafés - com jovens da Somália que chegaram à Finlândia por volta de 1994 e que freqüentam escolas finlandesas nos subúrbios de Helsinki. Meu interesse geral nesta pesquisa longitudinal era conhecer as experiências de passagem para a vida adulta em contextos altamente diferenciados, não apenas do ponto de vista do país anfitrião, mas também cultural e transnacionalmente. O tema, testes de DNA, toca na questão central desta pesquisa de modo profundo. Aqui, crescer não é visto como uma simples questão biológica. É um processo social no qual as relações, como laços de parentesco, são constituídas, vivenciadas e contestadas. Essas relações são poderosas para a identificação individual e social. A testagem-DNA pode violar simbólica e fisicamente o processo social de identificações íntimas e de integridade pessoal.The central themes of this article, family reunification in general, and DNA testing in particular, came to the fore during a research project about young Somalians in Finland. Since 1996, I have been conducting ethnographic research - in schools, youth clubs, streets and cafés - with youngsters from Somalia who arrived in Finland around 1994, and who attend Finnish schools in the suburbs of Helsinki. My general interest in this longitudinal study was to learn about the experiences of coming of age in highly dispersed settings, not only in the vein of a local host country, but also culturally and transnationally. Here, growing up is seen not as a simple biological question. It is a social process in which relationships such as kinship ties are constituted, experienced, and contested. These are powerful relations for individual and social

  7. Profiling of virulence associated genes of Pasteurella multocida isolated from cattle.

    Science.gov (United States)

    Verma, Subhash; Sharma, Mandeep; Katoch, Shailja; Verma, Lovit; Kumar, Sandeep; Dogra, Vishal; Chahota, Rajesh; Dhar, Prasenjit; Singh, Geetanjali

    2013-03-01

    Pasteurella multocida is a causative agent of many major diseases of which haemorrhagic septiciemia (HS) in cattle & a buffalo is responsible for significant losses to livestock sector in India and south Asia. The disease outcome is affected by various host- and pathogen-specific determinants. Several bacterial species-specific putative virulence factors including the capsular and virulence associated genes have been proposed to play a key role in this interaction. A total of 23 isolates of P. multocida were obtained from 335 cases of various clinically healthy and diseased cattle. These isolates were examined for capsule synthesis genes (capA, B, D, E and F) and eleven virulence associated genes (tbpA, pfhA, toxA, hgbB, hgbA, nanH, nanB, sodA, sodC, oma87 and ptfA) by PCR. A total of 19 P. multocida isolates belonging to capsular type B and 4 of capsular type A were isolated. All isolates of capsular type B harboured the virulence associated genes: tbpA, pfhA, hgbA, sodC and nanH, coding for transferrin binding protein, filamentous hemagglutinin, haemoglobin binding protein, superoxide dismutase and neuraminidases, respectively; while isolates belonging to capsular type A also carried tbpA, pfhA, hgbA and nanH genes. Only 50 % of capsular type A isolates contained sodC gene while 100 % of capsular type B isolates had sodC gene. The gene nanB and toxA were absent in all the 23 isolates. In capsular type A isolates, either sodA or sodC gene was present & these genes did not occur concurrently. The presence of virulence associated gene ptfA revealed a positive association with the disease outcome in cattle and could therefore be an important epidemiological marker gene for characterizing P. multocida isolates.

  8. Correlação da expressão do gene CXCL12 com o perfil de metilação de ilhas de CpG na região promotora em linhagens tumorais de mama

    OpenAIRE

    Klassen, liliane Maria Bacaro

    2013-01-01

    Resumo: O câncer de mama é o tipo mais comum e a principal causa de morte por câncer entre as mulheres no mundo todo. As metástases contribuem com 90% das causas de morte por câncer. O estudo de novos marcadores moleculares de câncer estão em amplo e atual desenvolvimento, tanto para diagnóstico como prognóstico da doença. Diversos estudos tem mostrado o papel de eventos epigenéticos entre eles a metilação do DNA, em regiões promotoras de genes como um evento importante no processo de tumorig...

  9. The Na+ transport in gram-positive bacteria defect in the Mrp antiporter complex measured with 23Na nuclear magnetic resonance.

    Science.gov (United States)

    Górecki, Kamil; Hägerhäll, Cecilia; Drakenberg, Torbjörn

    2014-01-15

    (23)Na nuclear magnetic resonance (NMR) has previously been used to monitor Na(+) translocation across membranes in gram-negative bacteria and in various other organelles and liposomes using a membrane-impermeable shift reagent to resolve the signals resulting from internal and external Na(+). In this work, the (23)Na NMR method was adapted for measurements of internal Na(+) concentration in the gram-positive bacterium Bacillus subtilis, with the aim of assessing the Na(+) translocation activity of the Mrp (multiple resistance and pH) antiporter complex, a member of the cation proton antiporter-3 (CPA-3) family. The sodium-sensitive growth phenotype observed in a B. subtilis strain with the gene encoding MrpA deleted could indeed be correlated to the inability of this strain to maintain a lower internal Na(+) concentration than an external one. Copyright © 2013 Elsevier Inc. All rights reserved.

  10. Determination of the frequency of polymorphisms in genes related to the genome stability maintenance of the population residing at Monte Alegre, PA (Brazil) municipality; Determinacao da frequencia de polimorfismos em genes relacionados a manutencao da estabilidade do genoma na populacao residente no municipio de Monte Alegre, PA

    Energy Technology Data Exchange (ETDEWEB)

    Hozumi, Cristiny Gomes

    2010-07-01

    The human exposure to ionizing radiation coming from natural sources is an inherent feature of human life on earth, for man and all living things have always been exposed to these sources. Ionizing radiation is a known genotoxic agent which can affect the genomic stability and genes related to DNA repair may play a role when they have committed certain polymorphism. This study aimed to analyze the frequency of polymorphisms (SNPs) in genes of DNA repair and cell cycle control: hOGG1 (Ser326Cys), XRCC3 (Thr241 Met) and p53 (Arg72Pro) in saliva samples from a population located Monte Alegre, state of Para were collected in August 2008 and 40 samples of men and 46 samples of women, adding a total of 86 samples. By RFLP was determined the frequency of homozygous genotypes and / or heterozygous for polymorphic genes. The I)OGG1 gene was 5% of the allele 326Cys, XRCC3 gene found about 21 % of the allele 241 Met and p53 gene showed 40.8% of the 72Pro allele. And the genotype frequencies of individuals for the three genes were 91.04%, 88.06% and 59.7% for homozygous wild genotype, 5.97%, 11.94% and 22.39% for heterozygote genotype and 2,99%, zero and 17:91% for homozygous polymorphic hOGG1 genes respectively, XRCC3, p53. These values are similar to those found in previous studies. The influence of these polymorphisms, which are involved in DNA repair and consequent genotoxicity induced by radiation depends on dose and exposure factors such as smoking, which is statistically a factor in public health surveillance in the region. This study gathered information and molecular epidemiology in Monte Alegre, that help to characterization of local population. (author)

  11. O JORNALISTA NA PESQUISA E NA ARTE

    Directory of Open Access Journals (Sweden)

    Beatriz Marocco

    2013-06-01

    Full Text Available O jornalista tem sido objeto das artes e das ciências. Nesta incisão no tema do painel “Configurações e perspectivas da pesquisa em jornalismo no Brasil em diálogo com os estudos latinoamericanos em jornalismo”, realizado no Encontro Nacional da Sociedade Brasileira de Pesquisadores em Jornalismo/SBPJor (Curitiba, 10/11/2012, reconheci que o âmbito acadêmico projeta dois tipos de ação: em rede de pesquisadores de diferentes países e regiões do mundo, com participação de países da América Latina, na pesquisa colaborativa e em projetos de investigação com outras bases de colaboração. Ambas as modalidades dialogam com a produção do cinema e da literatura – que visibiliza a grande potência do profissional para tramas e tensões. 

  12. Avaliação da superexpressão da proteína p53 e das mutações no éxon 8 do gene TP53 em carcinomas mamários caninos e glândulas normais

    Directory of Open Access Journals (Sweden)

    Maria Juliana D. Teixeira

    2011-06-01

    Full Text Available Este estudo foi realizado com o objetivo de avaliar a expressão da proteína p53, pela técnica de imuno-histoquímica, em neoplasmas mamários malignos em cadelas, além de investigar mutações no éxon 8 do gene supressor Tp53 por meio do padrão de bandas obtidas por PCR-RFLP. Dezenove mamas de cadelas saudáveis foram usadas como controle (Grupo 1. Amostras de 18 casos de tumores malignos (Grupo 2 e suas glândulas mamárias contralaterais (Grupo 3 foram obtidas na rotina do Hospital Veterinário da UFRPE. Os tumores foram identificados histologicamente e classificados em graus de malignidade. O método da estreptoavidina-biotina peroxidase foi utilizado para a análise da expressão de p53 por imuno-histoquímica, de acordo com a localização e intensidade da coloração. A expressão da proteína p53 não foi observada nas amostras do Grupo 1, mas foi encontrada em todas as amostras de tumores malignos (Grupo 2 seja só no núcleo, ou também no citoplasma. No Grupo 3, a expressão foi observada em quatro amostras normais e em duas que apresentavam tumor. Para a análise molecular, o DNA genômico foi extraído e submetido à PCR-RFLP com as seguintes endonucleases: AluI, BsoBI, DdeI e SmaI. O padrão de bandas foi polimórfico entre os grupos, mas não entre as variantes tumorais. Esse polimorfismo detectou mutações no fragmento estudado - éxon 8 do gene Tp53 - que podem resultar em alterações nos nucleotídeos, localizados nos sítios de restrição das enzimas. Esses achados levam a conclusão de que a imunoexpressão da p53 não tem relação com o subtipo histológico ou grau de malignidade do tumor, mas sim com a presença dos tumores no tecido mamário de cadelas. A PCR-RFLP pode ser usada como importante ferramenta para o estudo da carcinogênese mamária na cadela, possibilitando gerar diagnósticos precoces através do polimorfismo obtido com endonucleases de restrição pré-selecionadas.

  13. Romanskoe vlijanie na staroserbskij sintaksis

    OpenAIRE

    Pavlović Slobodan

    2010-01-01

    V nastojaščej rabote rassmatrivajutsja kontekstual'nye aspekty vlijanija romanskogo substrata i adstrata na staroserbskij sintaksis. Na sintaksis staroserbskogo razgovornogo jazyka mogli vlijat' vlašskie, dalmatinskie i ital'janskie dialekty. Na sintaksis staroserbskoj pis'mennosti (krome serbsko-slavjanskogo jazyka i romanskogo substrata) okazali vlijanie latinskij i ital'janskij jazyki, poskol'ku na territorii rasprostranenija serbskogo jazyka peresekalis' dve velikie kul'tury: Pax Sl...

  14. Clinical and serial MRI findings of a sialidosis type I patient with a novel missense mutation in the NEU1 gene.

    Science.gov (United States)

    Sekijima, Yoshiki; Nakamura, Katsuya; Kishida, Dai; Narita, Aya; Adachi, Kaori; Ohno, Kosaku; Nanba, Eiji; Ikeda, Shu-Ichi

    2013-01-01

    The case of a Japanese sialidosis type I patient with a novel NEU1 gene mutation is described. The patient developed an unsteady gait at age 14 and was referred to our hospital at age 16. On admission, subnormal intelligence, dysarthria, myoclonus, intentional tremors, limb and gait ataxia, hyperreflexia and macular cherry-red spots were observed. An enzymological analysis revealed a primary deficiency of neuraminidase. An NEU1 gene analysis identified two heterozygous missense mutations: p.P80L and p.D135N. The p.D135N mutation is a novel mutation that is considered to be associated with the mild clinical phenotype of sialidosis. Serial brain MRI showed diffuse brain atrophy progressing rapidly over the 41-month observation period.

  15. Preparing for NA4

    CERN Multimedia

    CERN PhotoLab

    1977-01-01

    Here, in one of the EF workshop, Albert Duchêne works on a pretty piece of mechanics. A few others await to be attended on the left. There are indications that the pipes were meant to house the carbon target (subdivided in eight sections) to be installed inside the toroid magnet of the NA4 experiment. The external strips were designed to possibly correct the magnetic field (???).

  16. Exclusion of NEU1 and PPGB from candidate genes for a lysosomal storage disease in Japanese Black cattle.

    Science.gov (United States)

    Masoudi, Ali Akbar; Yamato, Osamu; Yoneda, Kazuhiro; Tsuji, Takehito; Mikami, Osamu; Kunieda, Tetsuo

    2009-10-01

    A case of lysosomal storage disease has been reported in a calf of Japanese Black cattle. Lysosomal storage diseases are hereditary diseases caused by deficiency of lysosomal hydrolases. The clinical and pathological features and accumulated substrates of the affected animal indicated a possibility of sialidosis or galactosialidosis caused by deficiency of neuraminidase (NEU1) or protective protein for beta-galactosidase (PPGB). In the present study, we investigated nucleotide sequences of the genes encoding these two proteins to evaluate whether mutation of these genes is involved in this disease. We determined cattle genomic sequences of these two genes by using bovine EST sequences and the nucleotide sequences of all exons of these genes were compared between affected and normal animals. The results showed several nucleotide substitutions, but none of them was a functional mutation or specific to the affected animal. Furthermore, genotyping of the microsatellite markers in the vicinity of these two genes revealed no homozygosity of the chromosomal regions including these genes in the affected animal. These findings indicated that neither NEU1 nor PPGB gene is responsible for the lysosomal storage disease of Japanese Black cattle and therefore the disease is neither sialidosis nor galactosialidosis.

  17. Terapia gênica com VEGF para angiogênese na angina refratária: ensaio clínico fase I/II VEGF gene therapy for angiogenesis in refractory angina: phase I/II clinical trial

    Directory of Open Access Journals (Sweden)

    Renato A. K. Kalil

    2010-09-01

    Full Text Available OBJETIVO: Avaliar a segurança, viabilidade e efeitos iniciais, clínicos e sobre a perfusão miocárdica, da administração intramiocárdica, transtorácica, de VEGF 165 plasmidial em pacientes com doença arterial coronariana avançada e angina refratária, não passíveis de revascularização percutânea e cirúrgica. MÉTODOS: Ensaio clínico fase I/II. Treze pacientes cardiopatas isquêmicos com angina refratária apesar de tratamento medicamentoso máximo por no mínimo seis meses, não passíveis de revascularização cirúrgica ou por cateter foram submetidos a injeções intramiocárdicas de 2000µg VEGF 165 plasmidial. Os pacientes foram avaliados por cintilografia miocárdica, teste ergométrico, questionário de qualidade de vida (Minnesota e determinação das classes de insuficiência cardíaca (NYHA e angina (CCS. RESULTADOS: Não houve óbitos ou reintervenções. Durante o período de tratamento medicamentoso máximo, não se observou diferenças em cintilografias miocárdicas, testes ergométricos e questionários de qualidade de vida, ainda, houve tendência a piora das classes NYHA (P=0,05 e CCS (P=0,05. Três meses após intervenção, observou-se melhora dos escores cintilográficos SSS (18,38±7,51 vs. 15,31±7,29, P=0,003 e SRS (11,92±7,49 vs. 8,53±6,68, P=0,002, porém não na proporção da extensão da área de miocárdio isquêmico (23,38±13,12% vs. 20,08±13,88%, P=0,1. Houve tendência a melhora dos METs nas ergometrias (7,66±4,47 vs. 10,29±4,36, P=0,08, melhora do escore de qualidade de vida (48,23±18,35 vs. 30,15±20,13; P=0,02 e das classes NYHA (3,15±0,38 vs. 1,77±0,83, P=0,001 e CCS (3,08±0,64 vs. 1,77±0,83, P=0,001, no mesmo período. CONCLUSÕES: A terapia demonstrou-se segura e viável nesta série de pacientes. Os resultados iniciais tendem a demonstrar melhora na gravidade da angina e redução da intensidade da isquemia miocárdica.OBJECTIVE: Safety, feasibility and early myocardial angiogenic

  18. Hidroma subdural na fossa posterior

    Directory of Open Access Journals (Sweden)

    José Carlos Vasques

    1970-03-01

    Full Text Available Os autores relatam um caso de hidroma subdural na fossa craniana posterior conseqüente a traumatismo na região occipital. O paciente foi operado com pleno sucesso. A raridade da localização de hidroma na fossa posterior é salientada, sendo discutidos os possíveis mecanismos etio-patogênicos.

  19. Effect of the kappa-casein gene polymorphism, breed and seasonality on physicochemical characteristics, composition and stability of bovine milk Efeito do polimorfismo do gene da kappa-caseína, da raça e da sazonalidade sobre as características físico-químicas, de composição e de estabilidade do leite bovino

    Directory of Open Access Journals (Sweden)

    Bruno Garcia Botaro

    2009-12-01

    Full Text Available The objective of this study was to evaluate the effect of genetic polymorphism of kappa-casein, breed and seasonality on the physicochemical characteristics, composition and stability of milk in commercial dairy herds. A total of 879 milk and blood samples were collected from 603 Holstein and 276 Girolando cows, obtained during rainy and dry seasons. Milk samples were analyzed to determine the physicochemical characteristics, composition and ethanol stability, while blood samples were subjected to polymerase chain reaction to identify the kappa-casein genotype. The frequencies of genotypes AA, AB and BB of k-casein were respectively, 66.83, 31.84 and 1.33% for Holstein, and 71.38, 27.90 and 0.72% for the Girolando cows, respectively. The A allele was more frequent than the B allele, both for Holstein (0.827 and 0.173 and Girolando cows (0.853 and 0.147, respectively. Cows of AB and BB genotypes showed a higher milk fat content compared to the AA genotype. There was an interaction between breed and seasonality on the concentration of milk urea with higher values for Holstein and Girolando cows in the rainy and dry season, respectively. The levels of lactose, total solids, crude protein, true protein, casein and the casein:true protein ratio were higher during the dry season, while during the rainy season, the somatic cell count and milk urea concentration were higher. There was no association between milk stability and k-casein genotypes, but Holstein cows showed higher milk stability than Girolando cows, and milk was more stable during the rainy season than during the dry season.Objetivou-se avaliar o efeito do polimorfismo genético da kappa-caseína, da raça e da sazonalidade sobre as características físico-químicas, a composição e a estabilidade do leite bovino de rebanhos comerciais. Foram coletadas 879 amostras de leite e de sangue de 603 vacas da raça Holandesa e 276 da raça Girolando, obtidas durante as estações seca e chuvosa

  20. Sialyltransferase and Neuraminidase Levels/Ratios and Sialic Acid Levels in Peripheral Blood B Cells Correlate with Measures of Disease Activity in Patients with Systemic Lupus Erythematosus and Rheumatoid Arthritis: A Pilot Study.

    Directory of Open Access Journals (Sweden)

    Lieh-Bang Liou

    Full Text Available We attempted to determine whether the level of enzymes sialyltransferase (ST and neuraminidase (Neu and sialic acid (SIA in patients with systemic lupus erythematosus (SLE correlates with the SLE Disease Activity Index (SLEDAI and in patients with rheumatoid arthritis (RA correlates with the Disease Activity Score28 (DAS28.We examined cell-surface levels of ST6Gal-1, Neu1, ST3Gal-1, Neu3, α-2,6-SIA, and α-2,3-SIA by using fluorescent anti-enzyme antibodies, fluorescent-conjugated Sambucus nigra lectin, and fluorescent-conjugated Maackia amurensis lectin on blood cells in SLE and RA patients and assessed correlations of these levels with SLEDAI and with DAS28. Areas under the curve (AUC were calculated for different variables against SLEDAI.The B-cell ST3Gal-1/Neu3 ratio positively correlated with SLEDAI scores (ρ = 0.409 and P = 0.002, statistically significant after Bonferroni' correction for multiple analyses.. It was supported by the inverse correlation of B-cell Neu3 levels with SLEDAI scores (ρ = -0.264, P = 0.048. The B-cell ST3Gal-1/Neu3 ratio against SLEDAI yielded an AUC of 0.689, which was comparable to that of anti-dsDNA levels at 0.635. In contrast, both ST3Gal-1 and Neu3 levels of RA B cells (r = 0.376, P = 0.013; r = 0.425, P = 0.005, respectively correlated positively with high disease-activity DAS28 scores.B-cell ST3Gal-1/Neu3 ratios in SLE and B-cell ST3Gal-1 and Neu3 levels in RA with high disease-activity DAS28 scores correlated with disease activity measures and may be useful in monitoring disease activities.

  1. Isolation and preliminary function analysis of a Na + /H + antiporter ...

    African Journals Online (AJOL)

    genome. Northern blot analysis showed negligible expression of the gene in roots, but expression was detected in stems and leaves. To test the function of ... plants from NCBI revealed that MzNHX1 shared high identity (>86%) with vacuolar Na+/H+ antiporters, including RhNHX1 from rose, cNHX1 from citrus, and AtNHX1 ...

  2. Intracellular localization of Na + /H + antiporter from Malus zumi ...

    African Journals Online (AJOL)

    In this study, we examined the intracellular localization of the product of Na+/H+ antiporter gene (MzNHX1) cloned from Malus zumi. Analysis using yeast cells expressing a fusion protein of MzNHX1 and green fluorescent protein confirmed the localization of MzNHX1 on the tonoplast.

  3. Expression and purification of the central stalk subunits of Na + ...

    African Journals Online (AJOL)

    , NtpD and NtpG subunits. The aim of the present study was cloning and expression of these central stalk subunits of E. hirae V-type Na+-ATPase. Here we cloned the synthesized DNA fragments, corresponding to ntpC, ntpD and ntpG genes, ...

  4. Effect of adaptogenic preparations on Na(+/H(+-antiporter function in plasma membrane of corn root cells under salinity conditions

    Directory of Open Access Journals (Sweden)

    N. O. Kovalenko

    2014-10-01

    Full Text Available Salinity is a hard stress factor for plant organisms which negative effect is caused chiefly by sodium toxic for plants. Plant cells try to remove Na+ from their cytoplasm outside and to vacuolar space by secondary active Na+/H+-antiporters. Their functions can be intensified by gene engineering methods­ however we try do it with the help of non-toxic bio­active preparations. A comparison of their effect on the plasma membrane of Na+/H+-antiporters was carried out on corn seedling roots of Zea mays L. exposed at 0.1 M NaCl. Before we have established that Methyure used by seed pretreating possesses a high salt protective ability as against Ivine. It was found that without NaCl exposition Na+/H+-antiporter­ activity in root plasma membrane was nearly unnoticeable but increased slightly with seedling age. Methyure and Ivine did not influence its activity in control root seedling. One day 0.1 M NaCl exposition evoked a considerable increasing of Na+/H+-antiporter activity and its gene expression but these effects disappeared at 10 day NaCl exposition. Methyure use reinforced Na+/H+-antiporter activity and prolonged it at NaCl exposition without effect on its gene expression whereas Ivine effects on these indexes were insignificant. Obtained results showed that the salt protective capability of Methyure is connected with plasma membrane Na+/H+-antiporter activation which is realized on molecular level.

  5. [Effect of adaptogenic preparations on Na+/H+-antiporter function in plasma membrane of corn root cells under salinity conditions].

    Science.gov (United States)

    Kovalenko, N O; Bilyk, Zh I; Palladina, T O

    2014-01-01

    Salinity is a hard stress factor for plant organisms which negative effect is caused chiefly by sodium toxic for plants. Plant cells try to remove Na+ from their cytoplasm outside and to vacuolar space by secondary active Na+/H+-antiporters. Their functions can be intensified by gene engineering methods however we try do it with the help of non-toxic bioactive preparations. A comparison of their effect on the plasma membrane of Na+/H+-antiporters was carried out on corn seedling roots of Zea mays L. exposed at 0.1 M NaCl. Before we have established that Methyure used by seed pretreating possesses a high salt protective ability as against Ivine. It was found that without NaCl exposition Na+/H+-antiporter activity in root plasma membrane was nearly unnoticeable but increased slightly with seedling age. Methyure and Ivine did not influence its activity in control root seedling. One day 0.1 M NaCl exposition evoked a considerable increasing of Na+/H+-antiporter activity and its gene expression but these effects disappeared at 10 day NaCl exposition. Methyure use reinforced Na+/H+-antiporter activity and prolonged it at NaCl exposition without effect on its gene expression whereas Ivine effects on these indexes were insignificant. Obtained results showed that the salt protective capability of Methyure is connected with plasma membrane Na+/H+-antiporter activation which is realized on molecular level.

  6. The CERES / NA45 experiment

    CERN Multimedia

    Laurent Guiraud

    2000-01-01

    Ceres is one of the second generation heavy ion experiments at CERN's SPS. It is dedicated to the study of electron-positron pairs in relativistic nuclear collisions. NA45 is one of the seven experiments (NA44, NA45, NA49, NA50, NA52, WA97/NA57 and WA98) involved in CERN's Heavy Ion programme which provided evidence for the existence of a new state of matter, the quark-gluon plasma. In this state, quarks, instead of being bound up into more complex particles such as protons and neutrons, are liberated and roam freely. Theory predicts that this state must have existed at about 10 microseconds after the Big Bang, before the formation of matter as we know it today.

  7. na Perua

    Directory of Open Access Journals (Sweden)

    Jorge Carvalho

    1989-01-01

    Full Text Available Antes de se caracterizar como tentativa de ensaio ou de análise a qualquer coisa, o presente artigo tem, antes de mais nada, a pretensão de se constituir uma provocação ao debate acerca da questão do esporte na escola. A provocação parte basicamente do relato da minha experiência pessoal como Secretário da Educação do Município de Aracaju, cargo que exerci no período de 1º de janeiro de 1986 a 27 de janeiro de 1987.

  8. The Na+ transporter, TaHKT1;5-D, limits shoot Na+ accumulation in bread wheat

    KAUST Repository

    Byrt, Caitlin Siobhan

    2014-10-01

    Bread wheat (Triticum aestivum L.) has a major salt tolerance locus, Kna1, responsible for the maintenance of a high cytosolic K+/Na+ ratio in the leaves of salt stressed plants. The Kna1 locus encompasses a large DNA fragment, the distal 14% of chromosome 4DL. Limited recombination has been observed at this locus making it difficult to map genetically and identify the causal gene. Here, we decipher the function of TaHKT1;5-D, a candidate gene underlying the Kna1 locus. Transport studies using the heterologous expression systems Saccharomyces cerevisiae and Xenopus laevis oocytes indicated that TaHKT1;5-D is a Na+-selective transporter. Transient expression in Arabidopsis thaliana mesophyll protoplasts and in situ polymerase chain reaction indicated that TaHKT1;5-D is localised on the plasma membrane in the wheat root stele. RNA interference-induced silencing decreased the expression of TaHKT1;5-D in transgenic bread wheat lines which led to an increase in the Na+ concentration in the leaves. This indicates that TaHKT1;5-D retrieves Na+ from the xylem vessels in the root and has an important role in restricting the transport of Na+ from the root to the leaves in bread wheat. Thus, TaHKT1;5-D confers the essential salinity tolerance mechanism in bread wheat associated with the Kna1 locus via shoot Na+ exclusion and is critical in maintaining a high K+/Na+ ratio in the leaves. These findings show there is potential to increase the salinity tolerance of bread wheat by manipulation of HKT1;5 genes.

  9. gene structure, gene expression

    Indian Academy of Sciences (India)

    Primer 5.0 software. To adjust for RNA quality and diffe- rences in cDNA concentration, we amplified actin as an internal control with the following primers: PtActin-F (5′-TG. AAGGAGAAACTTGCGTAT-3′) and PtActin-R (5′-GCA. CAATGTTACCGTACAGAT-3′). These genes were ampli- fied from first-strand cDNA using ...

  10. Microdissecção e captura a laser na investigação do gene TP53 em tecidos incluídos em parafina Laser-capture microdissection for TP53 gene analysis in paraffin-embedded tissues

    Directory of Open Access Journals (Sweden)

    Shadia Muhammad Ihlaseh

    2007-02-01

    Full Text Available INTRODUÇÃO: Microdissecção e captura a laser (MCL é uma técnica de desenvolvimento recente que permite a coleta de células individuais ou pequeno conjunto de células para análise molecular. Atualmente, no Brasil, há raros microscópios para MCL, de modo que a divulgação dos procedimentos inerentes a essa técnica é oportuna para destacar seu amplo potencial para diagnóstico e investigação. OBJETIVO: Este trabalho descreve a padronização dos procedimentos de MCL e de extração de DNA de material fixado em formalina e incluído em parafina. MATERIAL E MÉTODOS: Foram estudados o éxon 8 do gene TP53 e o gene da ciclofilina em amostras de tecido normal e de neoplasias de fígado e rim provenientes de modelo de carcinogênese química induzida em rato. A extração do DNA foi comprovada por reação em cadeia da polimerase (nested-PCR. RESULTADOS: Foram padronizados os procedimentos de preparo dos cortes histológicos, de microdissecção e captura a laser e de obtenção de seqüências gênicas pela reação de nested-PCR para tecidos incluídos em parafina. Obtivemos amplificação de 48,3% das amostras para o éxon 8 do gene TP53 e 51,7% para o gene da ciclofilina. Considerando pelo menos um dos dois segmentos gênicos, foram amplificadas 79,3% das amostras. DISCUSSÃO E CONCLUSÃO: A extração de DNA de tecidos fixados em formalina e incluídos em parafina e a técnica de nested-PCR foram adequadamente padronizadas para produtos gênicos de interesse, obtidos de material coletado por MCL. Esses procedimentos podem ser úteis para a obtenção de seqüências de DNA de arquivos para análise molecular.BACKGORUND: Laser-capture micro-dissection (LCM is a recently developed procedure that provides single cells or specific cell groups for molecular analysis. Currently, there are few LCM systems in Brazil, in such a way that it is necessary to disseminate the technical procedures inherent to the methodology, and also to

  11. Comprometimento pulmonar na leptospirose

    Directory of Open Access Journals (Sweden)

    Jorge Eduardo Manhães de Carvalho

    1992-03-01

    Full Text Available Em 23 pacientes com leptospirose apresentando comprometimento pulmonar, internados no Hospital Universitário Antônio Pedro da XJFF, Niterói, hemoptise e hemoptóicos foram observados em 21,7% e 30,4%, respectivajnente. Gasometria arterial revelou hipoxemia e hipocapnia na maioria dos casos. Radiografia de tórax em 15 pacientes mostrou comprometimento alveolar em 60%, comprometimento intersticial-reticular em 6%, padrão misto (alveolar e intersticial em20%e ausência de alterações radiológicas em 14%. A necrópsiade 13 pacientes mostrou edema, congestão e hemorragia nos pulmões em 100% dos casos. A hemorragia foi focal em 46% e difusa em 54% dos casos. Houve formação de membrana hialina em 30% e trombos de fibrina em 46% dos pulmões estudados, o que estabelece o diagnóstico da coagulação intravascular disseminada e a ocorrência da síndrome de angústia respiratória na leptospirose.

  12. Vrapci na tlu

    Directory of Open Access Journals (Sweden)

    Ana Ille Horvat

    2017-12-01

    Full Text Available Princeze nikada ne umiru. Udaju se i žive sretno do kraja života. "Ideja će mi sinuti kad se najmanje budem nadala", kaže Alice, "uvijek je tako. Neću očajavati." Upravo sjedi na kauču držeći bilježnicu u rukama u koju je trenutak prije, velikim štampanim slovima na vrhu lista, upisala "ideje za Afonsov rođendan". Pažljivo je povukla pet strelica i svaku označila brojem. Poslije toga nije napisala više ništa. Ovo je već sedmi popis koji radi otkako je sjela. Pogleda uprta prema dnevnom boravku, oko kažiprsta lijeve ruke mota plamen jarkoplave kose, kao u lutke."Kako se to smišlja neka ideja", upita se šapatom koji je uvježbala još u mladosti, "kako?"

  13. Rare kaon decays (NA48/2 and NA62)

    OpenAIRE

    Goudzovski, Evgueni

    2013-01-01

    The rare decay $K^\\pm\\to\\pi^\\pm\\gamma\\gamma$ has been recently measured from data samples collected by the NA48/2 and NA62 experiments at CERN. These measurements are presented, including model-independent spectrum measurements and fits to the Chiral Perturbation Theory description. The rare decay $K^\\pm\\to\\pi^\\pm\\gamma\\gamma$ has been recently measured from data samples collected by the NA48/2 and NA62 experiments at CERN. These measurements are presented, including model-independent spec...

  14. In vitro expression of native H5 and N1 genes of avian influenza virus by using Green Fluorescent Protein as reporter

    Directory of Open Access Journals (Sweden)

    Risza Hartawan

    2011-10-01

    Full Text Available The hemagglutinin and neuraminidase are important immunogen of avian influenza virus that are suitable for recombinant experimentation. However, both genes have been experienced rapid mutation resulting in diverse variety of genotypes. Hence, gene expression in recombinant systems will be difficult to predict. The objective of the study was to examine expression level of H5 and N1 genes from a field isolate by cloning the genes into expression vector pEGFP-C1. Two clones respresenting fulllength of H5 and N1 gene in plasmid pEGFP-C1 were transfected into chicken embryo fibroblasts (CEF, rabbit kidney (RK13 and African green monkey kidney (VERO cells using Lipofectamine ‘Plus’ reagent. The experiment showed level of gene expression in the VERO cell was higher than in the RK13 and CEF cells. Observations using fluorescent microscopy and Western blotting revealed that the N1 gene was expressed better in all cells compared to the H5 gene.

  15. Role of divalent cations, pH, cytoskeleton componentes and surface charge on the adhesion of Trichomonas vaginalis to a polystyrene substrate Papel de cátions divalentes, pH, componentes de citoesqueleto e carga de superfície na adesão de Trichomonas vaginalis a um substrato de poliestireno

    Directory of Open Access Journals (Sweden)

    Fernando Costa e Silva Filho

    1987-09-01

    Full Text Available The process of adhesion of three different strains of Trichomonas vaginalis to a polystyrene substrate was analysed. The process of adhesion was dependent on the time of incubation and the pH of the phosphate-buffered solution (PBS in which the parasites were suspended. The highest indices of adhesion were observed after an incubation time of 60 min at pH 6.6. The adhesion index increased when the parasites were incubated in the presence of culture media or when Ca++ or Mg++ was added to the PBS solution, whereas cytochalasin B, trypsin or neuraminidase reduced adhesion. Incubation of the parasites in the presence of poly-L-lysine facilitated the process of adhesion. Incubation of the parasites or polystyrene beads in the presence of poly-L-lysine led to important changes in their surface charge.O processo de adesão de três cepas de Trichomonas vaginalis a um substrato de poliestireno foi estudado. Verificou-se que este processo depende do tempo de incubação e do pH da solução salina em que os parasitos se encontram. A maior taxa de adesão foi observada após 60 minutos de incubação a pH 6,6. A adesão é mior se Ca++ ou Mg++ for adicionado ao meio. Tratamento das células em citocalasina B, tripsina ou neuraminidase reduz a adesão enquanto tratamento com poli-L-lisina facilita esta adesão. Incubação dos parasitos ou esferas de poliestireno na presença de poli-L-lisina provoca alterações importantes na carga de superfície.

  16. Role of wild type p53 and double suicide genes in interventional therapy of liver cancer in rabbits Papel do p53 selvagem e do duplo genes suicidas na terapia intervencionista do câncer de fígado em coelhos

    Directory of Open Access Journals (Sweden)

    Hong-xin Niu

    2012-08-01

    Full Text Available PURPOSE: To investigate the feasibility of interventional lipiodol embolism and multigene therapy in combination with focal chemotherapy in the treatment of VX2 liver cancer in rabbits. METHODS: Forty five rabbits with cancer larger than 2cm in diameter were randomly divided into five groups (n=9 per group. In Group 1, animals were treated with 0.9% sodium chloride. In Group 2, animals received lipiodol embolism. In Group 3, animals received lipiodol embolism and p53 gene therapy. In Group 4, animals received lipiodol embolism and TK/CD gene therapy. In Group 5, animals received lipiodol embolism and p53 and TK/CD gene therapy. Ultrasonography and CT were performed before and at ten days after interventional therapy. RESULTS: The VX2 model of liver cancer was successfully established in rabbits and interventional therapy smoothly performed. At ten days after interventional therapy, significant difference in the tumor volume was noted among five groups (pOBJETIVO: Investigar a possibilidade de terapia multigênica e intervenção por embolização com lipiodol em combinação com quimioterapia focal no tratamento de câncer de fígado VX2 em coelhos. MÉTODOS: Quarenta e cinco coelhos com câncer maior do que 2cm de diâmetro foram distribuídos, aleatoriamente, em cinco grupos (n=9 por grupo. Grupo 1: animais foram tratados com cloreto de sódio 0,9% e no grupo 2 os animais receberam embolização com lipidol. Grupo 3: animais receberam embolização com lipiodol e terapia do gene p53 e grupo 4 animais receberam embolização com lipiodol e terapia do gene TK/CD. Grupo 5: animais receberam embolização com lipiodol e terapia do gene p53 e do gene TK/CD. Ultrassonografia e tomografia computadorizada foram realizadas antes e dez dias após a intervenção terapêutica. RESULTADOS: O modelo VX2 de câncer de fígado foi estabelecido com sucesso em coelhos e a terapia intervencionista foi bem executada. Dez dias após a intervenção terap

  17. Polymorphisms of candidate genes associated with meat quality and ...

    African Journals Online (AJOL)

    No mutation was found in the RFLP-fragment of the FUT1 gene of four indigenous pig breeds by sequencing, while in the BPI gene two mutations were detected in the Tap Na breed. The resistant alleles of the FUT1 and BPI genes in the exotic breeds were significantly higher than those of indigenous pig breeds. Among the ...

  18. NA48 prototype calorimeter

    CERN Multimedia

    1990-01-01

    This is a calorimeter, a detector which measures the energy of particles. When in use, it is filled with liquid krypton at -152°C. Electrons and photons passing through interact with the krypton, creating a shower of charged particles which are collected on the copper ribbons. The ribbons are aligned to an accuracy of a tenth of a millimetre. The folding at each end allows them to be kept absolutely flat. Each shower of particles also creates a signal in scintillating material embedded in the support disks. These flashes of light are transmitted to electronics by the optical fibres along the side of the detector. They give the time at which the interaction occurred. The photo shows the calorimeter at NA48, a CERN experiment which is trying to understand the lack of anti-matter in the Universe today.

  19. O real na psicose

    Directory of Open Access Journals (Sweden)

    Maurício Castejón Herrmann

    2004-06-01

    Full Text Available Este artigo discute a noção de real na psicose, a partir das formulações de Lacan sobre o tema, presentes no Seminário 3 - As Psicoses, no Seminário 20 - Mais, Ainda e no texto Televisão. Considerando-se que a concepção de real tem uma indicação clínica, a hipótese que se formula é a de que a noção de real trabalhada no Seminário 20, Mais, Ainda representa uma continuidade da concepção de real desenvolvida no Seminário 3, As Psicoses

  20. Na Cauda do Cometa

    Science.gov (United States)

    Voelzke, M. R.

    2009-01-01

    Quando viam um cometa, os antigos gregos imaginavam uma estrela com uma vasta cabeleira. Não à toa, a palavra deriva do termo koma, que significa cabelo. Constituídos por fragmentos de gelo e gases, os cometas possuem um núcleo sólido, que pode ter vários quilômetros de diâmetro, e uma cauda que sempre aponta na direção contrária ao Sol, devido aos ventos solares. Graças à aparência de pontos luminosos em movimento (ao contrário de outros astros, que parecem estáticos), esses corpos celestes foram interpretados por diferentes povos com muito misticismo, inspirando mitos tanto de boas-novas como de maus presságios. Conheça algumas dessas histórias:

  1. Athéna

    OpenAIRE

    Camps, G.

    2013-01-01

    Dès l’époque archaïque le culte d’Athéna fut important à Cyrène au point qu’on a pu se demander si cette importance ne s’expliquait pas par l’existence d’une divinité libyque qui aurait été identifiée à la déesse guerrière et industrieuse. A l’appui de cette opinion, on peut retenir plusieurs données de qualités diverses. Il faut citer en premier lieu la déesse égypto-libyque Nît, très ancienne mais particulièrement adorée durant l’époque saïte, au moment où la Basse-Egypte est soumise à une ...

  2. Genome Sequence of a Reassortant H5N1 Avian Influenza Virus Isolated from Domestic Green-Winged Teal.

    Science.gov (United States)

    Xiong, Chaochao; Liu, Qian; Chen, Quanjiao; Chen, Jianjun

    2013-08-15

    An avian influenza virus strain, A/domestic green-winged teal/Hunan/3450/2006(H5N1) (DGW-T3450), was isolated from domestic green-winged teals. Genome analysis demonstrated that DGW-T3450 is a novel reassortant strain. The hemagglutinin (HA) and neuraminidase (NA) genes of this strain originated from H5N1 viruses circulating in poultry, while its remaining genes are derived from multiple ancestors, including viruses like those that infect wild birds.

  3. Comprometimento pulmonar na leptospirose

    Directory of Open Access Journals (Sweden)

    Jorge Eduardo Manhães de Carvalho

    1992-03-01

    Full Text Available Em 23 pacientes com leptospirose apresentando comprometimento pulmonar, internados no Hospital Universitário Antônio Pedro da XJFF, Niterói, hemoptise e hemoptóicos foram observados em 21,7% e 30,4%, respectivajnente. Gasometria arterial revelou hipoxemia e hipocapnia na maioria dos casos. Radiografia de tórax em 15 pacientes mostrou comprometimento alveolar em 60%, comprometimento intersticial-reticular em 6%, padrão misto (alveolar e intersticial em20%e ausência de alterações radiológicas em 14%. A necrópsiade 13 pacientes mostrou edema, congestão e hemorragia nos pulmões em 100% dos casos. A hemorragia foi focal em 46% e difusa em 54% dos casos. Houve formação de membrana hialina em 30% e trombos de fibrina em 46% dos pulmões estudados, o que estabelece o diagnóstico da coagulação intravascular disseminada e a ocorrência da síndrome de angústia respiratória na leptospirose.Tostudy thepulmonary complications in leptospirosis case records of 23 such patients admitted at the Hospital Universitário Antônio Pedro, Universidade Federal Fluminense, Niterói, Brasil, were reviewed. Hemoptysis were seen in 21.7% and sputal blood in 30.4% of patients. Arterial gasometry detected hypoxemia and hypocapnia in most cases. Thoracic radiology showed an alveolar pattern in 60% of the patients, alveolo-interstitial in 20%, interstitial in 6%, and in 14% the lungs were considered to be normal Necropsy of 13 cases showed edema, congestion and hemorrhage in the lungs in all cases. Hyaline membrane was found in 30% and fibrin thrombi in 46% of these cases, resulting in a diagnosis of adult respiratory distress syndrome and acute disseminated intravascular coagulation (consumption coagulopathy in leptospirosis.

  4. Construction and characterization of a glycoprotein E deletion mutant of bovine herpesvirus type 1.2 strain isolated in Brazil Construção e caracterização de uma amostra de BoHV-1.2 isolada no Brasil com uma deleção no gene da glicoproteína E

    Directory of Open Access Journals (Sweden)

    Ana C. Franco

    2002-09-01

    Full Text Available This paper describes the construction and characterization of a Brazilian strain of bovine herpesvirus type 1.2a (BoHV-1.2a with a deletion of the glycoprotein E (gE gene. The deletion was introduced by co-transfection of a deletion fragment containing the 5´and 3´gE flanking regions and genomic DNA of wild type BoHV-1 into bovine cells. Isolation of gE deletion mutant was performed by immunoperoxidase staining with an anti-gE monoclonal antibody. Viral clones were plaque purified and further examined by restriction endonuclesase digestion and Southern blot hybridization. This gE deletion mutant will be evaluated as a vaccinal virus, in order to determine its potential use for a differential vaccine.Este artigo descreve a construção e caracterização de uma amostra de um herpesvírus bovino tipo 1.2a (BoHV-1.2a que apresenta uma deleção na região genômica que codifica a glicoproteína E (gE. A deleção gênica foi induzida através da co-transfecção de um fragmento de deleção, contendo as regiões 5´e 3´flanqueadoras da gE, com o DNA viral intacto de uma amostra viral isolada de um animal que apresentava doença respiratória. O isolamento do vírus gE negativo (gE- foi realizado com auxílio da técnica de imunoperoxidase em que foi utilizado como anticorpo primário um anticorpo monoclonal anti-gE. O vírus gE- foi purificado e o DNA isolado desta amostra foi examinado através das técnicas de análise por enzimas de restrição e "Southern blot". Esta amostra gE- será avaliada como candidata para compor uma vacina diferencial contra a rinotraqueíte infecciosa dos bovinos.

  5. A ENGENHARIA NA ESCOLA

    Directory of Open Access Journals (Sweden)

    Valéria Pelizzer Casara

    2015-04-01

    Full Text Available Este projeto teve por objetivo principal mostrar às jovens do ensino médio, como as áreas de exatas e de engenharia podem estar inseridas em situações rotineiras por elas vivenciadas e incentivá-las para ingressarem nessas áreas. O projeto foi executado na escola de ensino Médio Nelson Horostecki na cidade de Chapecó, Santa Catarina. Mostrou-se de forma simples que as áreas de exatas e de engenharia têm aplicações práticas nas nossas vidas. No desenvolvimento do projeto, realizaram-se oficinas na escola cujo tema principal abordado foi o processo de fabricação do chocolate, visto que o mesmo é um produto muito apreciado pelas adolescentes. Dentro dessa temática, foram trabalhadas as questões que envolvem conhecimentos das áreas de exatas e de engenharia, mais especificamente a Engenharia Ambiental. O projeto auxiliou na formação cidadã e também na escolha profissional das meninas. Muitas das alunas participantes nunca haviam tido contato direto com alunos graduandos de cursos na área das exatas. Mais ainda, este projeto oportunizou a aproximação da universidade com a comunidade externa. Palavras-chave: Extensão Universitária, Ciências Exatas e Engenharia, Chocolate, Oficinas.   Engineering in the school Abstract: The main objective of this project was to show high school female students how the exact sciences and engineering can be included in their routine situations encouraging them to entering in these areas. The project was carried out at the Nelson Horostecki High School in Chapecó, Santa Catarina State, Brazil. It was showed in a simple manner how exact sciences and engineering have practical applications in our lives. Workshops were conducted in the school during the execution of the project in which the main subject was the chocolate manufacturing process, since chocolate is a product greatly appreciated by the young female people. Within this matter, the activities were performed using the knowledge of

  6. Kaon experiments at CERN: NA48 and NA62

    CERN Document Server

    INSPIRE-00293758

    2012-01-01

    Searches for violation of lepton flavour universality and lepton number conservation in kaon decays by the NA62 and NA48/2 experiments at CERN, status and future plans of the CERN kaon programme are presented. A precision measurement of the helicity-suppressed ratio $R_K$ of the $K^\\pm\\to e^\\pm\

  7. Regulation of the cardiac Na+ channel NaV1.5 by post-translational modifications.

    Science.gov (United States)

    Marionneau, Céline; Abriel, Hugues

    2015-05-01

    The cardiac voltage-gated Na(+) channel, Na(V)1.5, is responsible for the upstroke of the action potential in cardiomyocytes and for efficient propagation of the electrical impulse in the myocardium. Even subtle alterations of Na(V)1.5 function, as caused by mutations in its gene SCN5A, may lead to many different arrhythmic phenotypes in carrier patients. In addition, acquired malfunctions of Na(V)1.5 that are secondary to cardiac disorders such as heart failure and cardiomyopathies, may also play significant roles in arrhythmogenesis. While it is clear that the regulation of Na(V)1.5 protein expression and function tightly depends on genetic mechanisms, recent studies have demonstrated that Na(V)1.5 is the target of various post-translational modifications that are pivotal not only in physiological conditions, but also in disease. In this review, we examine the recent literature demonstrating glycosylation, phosphorylation by Protein Kinases A and C, Ca(2+)/Calmodulin-dependent protein Kinase II, Phosphatidylinositol 3-Kinase, Serum- and Glucocorticoid-inducible Kinases, Fyn and Adenosine Monophosphate-activated Protein Kinase, methylation, acetylation, redox modifications, and ubiquitylation of Na(V)1.5. Modern and sensitive mass spectrometry approaches, applied directly to channel proteins that were purified from native cardiac tissues, have enabled the determination of the precise location of post-translational modification sites, thus providing essential information for understanding the mechanistic details of these regulations. The current challenge is first, to understand the roles of these modifications on the expression and the function of Na(V)1.5, and second, to further identify other chemical modifications. It is postulated that the diversity of phenotypes observed with Na(V)1.5-dependent disorders may partially arise from the complex post-translational modifications of channel protein components. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Antigenicity of the 2015-2016 seasonal H1N1 human influenza virus HA and NA proteins.

    Directory of Open Access Journals (Sweden)

    Amelia M Clark

    Full Text Available Antigenic drift of the hemagglutinin (HA and neuraminidase (NA influenza virus proteins contributes to reduced vaccine efficacy. To analyze antigenic drift in human seasonal H1N1 viruses derived from the 2009 pandemic H1N1 virus (pH1N1-like viruses accounts for the limited effectiveness (around 40% of vaccination against pH1N1-like viruses during the 2015-2016 season, nasal washes/swabs collected from adult subjects in the Rochester, NY area, were used to sequence and isolate the circulating viruses. The HA and NA proteins from viruses circulating during the 2015-2016 season encoded eighteen and fourteen amino acid differences, respectively, when compared to A/California/04/2009, a strain circulating at the origin of the 2009 pandemic. The circulating strains belonged to subclade 6B.1, defined by HA amino acid substitutions S101N, S179N, and I233T. Hemagglutination-inhibiting (HAI and HA-specific neutralizing serum antibody (Ab titers from around 50% of pH1N1-like virus-infected subjects and immune ferrets were 2-4 fold lower for the 2015-2016 circulating strains compared to the vaccine strain. In addition, using a luminex-based mPlex HA assay, the binding of human sera from subjects infected with pH1N1-like viruses to the HA proteins from circulating and vaccine strains was not identical, strongly suggesting antigenic differences in the HA protein. Additionally, NA inhibition (NAI Ab titers in human sera from pH1N1-like virus-infected subjects increased after the infection and there were measurable antigenic differences between the NA protein of circulating strains and the vaccine strain using both ferret and human antisera. Despite having been vaccinated, infected subjects exhibited low HAI Ab titers against the vaccine and circulating strains. This suggests that poor responses to the H1N1 component of the vaccine as well as antigenic differences in the HA and NA proteins of currently circulating pH1N1-like viruses could be contributing to

  9. Complete genome sequence of Fer-de-Lance Virus reveals a novel gene in reptilian Paramyxoviruses

    Science.gov (United States)

    Kurath, G.; Batts, W.N.; Ahne, W.; Winton, J.R.

    2004-01-01

    The complete RNA genome sequence of the archetype reptilian paramyxovirus, Fer-de-Lance virus (FDLV), has been determined. The genome is 15,378 nucleotides in length and consists of seven nonoverlapping genes in the order 3??? N-U-P-M-F-HN-L 5???, coding for the nucleocapsid, unknown, phospho-, matrix, fusion, hemagglutinin-neuraminidase, and large polymerase proteins, respectively. The gene junctions contain highly conserved transcription start and stop signal sequences and tri-nucleotide intergenic regions similar to those of other Paramyxoviridae. The FDLV P gene expression strategy is like that of rubulaviruses, which express the accessory V protein from the primary transcript and edit a portion of the mRNA to encode P and I proteins. There is also an overlapping open reading frame potentially encoding a small basic protein in the P gene. The gene designated U (unknown), encodes a deduced protein of 19.4 kDa that has no counterpart in other paramyxoviruses and has no similarity with sequences in the National Center for Biotechnology Information database. Active transcription of the U gene in infected cells was demonstrated by Northern blot analysis, and bicistronic N-U mRNA was also evident. The genomes of two other snake paramyxovirus genotypes were also found to have U genes, with 11 to 16% nucleotide divergence from the FDLV U gene. Pairwise comparisons of amino acid identities and phylogenetic analyses of all deduced FDLV protein sequences with homologous sequences from other Paramyxoviridae indicate that FDLV represents a new genus within the subfamily Paramyxovirinae. We suggest the name Ferlavirus for the new genus, with FDLV as the type species.

  10. Imagens e representações da enfermagem acerca do stress e sua influência na atividade laboral Imágenes y representaciones de la enfermería respecto del estrés y su influencia en la actividad laboral Nursing images and representations concerning stress and influence on work activity

    Directory of Open Access Journals (Sweden)

    Renata da Silva Hanzelmann

    2010-09-01

    Full Text Available Os objetivos deste estudo foram: identificar as representações acerca dos fatores desencadeadores do estresse, atribuídos pelos profissionais de enfermagem, na atividade laboral; e discutir a influência destes na sua atividade laboral. Optou-se por um estudo descritivo com abordagem qualitativa, para o qual foram utilizadas as premissas das Representações Sociais, tendo os estudos realizados como referencial de análise. Para coleta utilizou-se como instrumento entrevista semiestruturada e individual. A análise foi através da técnica de análise de conteúdo, a fim de buscar elementos para a compreensão das imagens como representações dos profissionais de enfermagem, o significado dos fatores estressantes, e sua influência na atividade laboral. A população estudada vive e convive com a falta de condições de trabalho, escassez de recursos materiais e humanos, e ainda com pessoal não treinado; o trabalhador sente-se insatisfeito, com fadiga mental e física - situações que podem propiciar o aparecimento do estresse no desempenho das atividades laborais.Los objetivos de este estudio apuntaron a identificar las representaciones acerca de los factores desencadenantes del estrés, atribuidos por los profesionales de la enfermería a la actividad laboral, y discutir la influencia de estos en la labor diaria. Se optó por un estudio descriptivo con abordaje cualitativo, para el cual fueron utilizadas las premisas de las Representaciones Sociales, teniendo como referencial de análisis los estudios realizados. Para la recolección de datos se utilizó como instrumento una entrevista semiestructurada e individual. El análisis se realizó a través de la técnica de análisis de contenido, a fin de buscar elementos para la comprensión de las imágenes como representaciones de los profesionales de enfermería, el significado de los factores estresores y su influencia en la actividad laboral. La población estudiada vive y convive con

  11. Identidade molecular dos fitoplasmas associados aos enfezamentos do tomateiro e da berinjela com base na análise do gene 16S rDNA Molecular identity of the phytoplasma associated to stunting of tomato and eggplant on the basis of analyses of the 16S rDNA

    Directory of Open Access Journals (Sweden)

    Ana Paula de Oliveira Amaral Mello

    2007-09-01

    Full Text Available Doenças de hortaliças de ocorrência no território brasileiro e em outras áreas do mundo têm sido associadas a diversos fitoplasmas. Na região de Piracicaba-SP e Bragança Paulista-SP, em plantas de tomate e berinjela foram observados sintomas típicos de enfezamento caracterizados por porte reduzido, clorose foliar, superbrotamento de ramos, desenvolvimento anormal do cálice, encurtamento de entre-nós, redução no tamanho de folhas, flores e frutos. Através de duplo PCR, utilizando os iniciadores R16 mF1/mR2 e R16 F2n/R2, fragmentos de DNA de 1,2 kb foram amplificados de amostras sintomáticas, demonstrando a presença de fitoplasma nos tecidos das plantas. O uso de iniciadores específicos demonstrou que estes fitoplasmas eram afiliados ao grupo 16SrIII. Análises de RFLP, usando as enzimas de restrição AluI, HpaII, KpnI, MboI, MseI e RsaI confirmaram que os fitoplasmas detectados eram representantes do grupo 16SrIII. Os fragmentos de DNA amplificados foram clonados em Escherichia coli, sequenciados e comparados, por homologia de seqüência, entre si e com outros fitoplasmas do grupo 16SrIII. Um índice de similaridade de seqüência acima de 95% foi encontrado quando seqüências dos fitoplasmas detectados em tomate e berinjela foram comparadas com aquelas de outros representantes do grupo 16SrIII. Um índice de 98-99% foi obtido quando seqüências dos fitoplasmas encontrados em tomate e berinjela foram comparadas entre si. Estes resultados evidenciaram que o enfezamento do tomateiro e da berinjela podem estar associados a um mesmo fitoplasma, com base na análise de seqüências do gene do 16S rDNA.Vegetable diseases occurring in the Brazilian territory and around the world have been associated with various phytoplasmas. In the region of Piracicaba-SP and Bragança-SP, in eggplant and tomato plants typical symptoms of stunting characterized by reduced canopy, leaf yellowing, proliferation of shoots, calix malformation

  12. Estudo comparativo entre um sistema de sonda genética e métodos clássicos na identificação das micobactérias Gene probes versus classical methods in the identification of mycobacteria

    Directory of Open Access Journals (Sweden)

    Andréa Gobetti Vieira Coelho

    2008-11-01

    Full Text Available OBJETIVO: O aparecimento da co-infecção tuberculose/HIV e o aumento de casos de doenças provocadas por micobactérias não-tuberculosas (MNT exigem repostas laboratoriais rápidas tanto no isolamento como na identificação das micobactérias. O objetivo deste trabalho foi avaliar a identificação das micobactérias através de sonda genética em comparação com os métodos bioquímicos clássicos. MÉTODOS: Entre 2002 e 2004, foram analisadas 178 culturas de micobactérias, confirmadas como bacilos álcool-ácido resistentes e obtidas de isolados clínicos de pacientes sintomáticos respiratórios ou com suspeita clínica de tuberculose pulmonar e/ou micobacterioses, atendidos nas Unidades de Saúde da Baixada Santista. RESULTADOS: A sonda genética identificou 137 amostras (77% como complexo Mycobacterium tuberculosis e 41 (23% como MNT. A discordância observada de 3% entre os métodos ocorreu apenas no ano de implantação (2002. Ao comparar os métodos, a sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo da sonda genética foram 98%, 93%, 98% e 93%, respectivamente. CONCLUSÕES: Apesar do custo elevado, a identificação de micobactérias pela técnica molecular é mais rápida: máximo de 3 h vs. 28-30 dias para os métodos clássicos. A utilização de sondas genéticas é uma técnica molecular validada, simples e disponível no mercado, com elevada especificidade, sensibilidade e rapidez, o que justifica sua implantação e uso rotineiro em laboratórios de referência, facilitando o diagnóstico e permitindo uma intervenção clínica ágil.OBJECTIVE: The emergence of tuberculosis/HIV co-infection and the increase in the number of cases of infection with nontuberculous mycobacteria (NTM require rapid laboratory test results in the isolation and identification of mycobacteria. The objective of this study was to evaluate the identification of mycobacteria by means of gene probes in comparison with

  13. Romanskoe vlijanie na staroserbskij sintaksis

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    Pavlović Slobodan

    2010-01-01

    Full Text Available V nastojaščej rabote rassmatrivajutsja kontekstual'nye aspekty vlijanija romanskogo substrata i adstrata na staroserbskij sintaksis. Na sintaksis staroserbskogo razgovornogo jazyka mogli vlijat' vlašskie, dalmatinskie i ital'janskie dialekty. Na sintaksis staroserbskoj pis'mennosti (krome serbsko-slavjanskogo jazyka i romanskogo substrata okazali vlijanie latinskij i ital'janskij jazyki, poskol'ku na territorii rasprostranenija serbskogo jazyka peresekalis' dve velikie kul'tury: Pax Slavia Orthodoxa i Pax Romana Catholica. Sintaksičeskoe vlijanie latinskogo i ital'janskogo jazykov na staro-serbskuju pis'mennost' osuščestvljalos' prežde vsego čerez zapadnye obrazcy delovoj perepiski. [Projekat Ministarstva nauke Republike Srbije: Istorija srpskog jezika

  14. Valores na escola

    Directory of Open Access Journals (Sweden)

    Menin Maria Suzana De Stefano

    2002-01-01

    Full Text Available Neste texto pretende-se discorrer sobre valores morais na escola e suas implicações para a formação de professores. Para tanto discutir-se-á, em primeiro lugar, e brevemente, o que são valores morais, ou éticos, e como a escola pode situar-se em relação a eles. Em seguida, serão relatadas algumas observações a respeito de valores de professores e práticas daí decorrentes. São comentados resultados de pesquisa que ilustram a transmissão de valores de forma doutrinal e a educação moral e cívica tal como realizada na ditadura militar, e, por outro lado, a posição relativista e/ou de laissez-faire que certas escolas podem adotar, metodologicamente, sobre a educação em valores. Finalmente, defender-se-á a idéia de que é necessária uma discussão sobre valores pelos diversos membros da escola e uma opção por uma metodologia para ensiná-los, seja os professores, em sua formação inicial e continuada, seja os alunos. A teoria de desenvolvimento moral de Jean Piaget será apresentada como uma referência possível para a educação em valores. Exemplos de situações escolares de conflito de valores entre direção, pais e alunos são discutidas para ilustrar como uma escola pode adotar um procedimento democrático de educação em valores, que se apresenta como um terceiro caminho possível de educação moral nas escolas, além das posições doutrinárias ou relativistas.

  15. O papel do gene e da molécula HLA-G na expressão clínica das doenças reumatológicas The role of the HLA-G gene and molecule on the clinical expression of rheumatologic diseases

    Directory of Open Access Journals (Sweden)

    Claiton Viegas Brenol

    2012-02-01

    Full Text Available O antígeno leucocitário humano G (HLA-G é uma molécula não clássica de complexo principal de histocompatibilidade (MHC de classe I, caracterizada por baixo polimorfismo em sua região codificadora, um padrão de distribuição tecidual limitado em condições fisiológicas e expressão por meio de isoformas solúveis e acopladas à superfície de membranas por meio de splicing alternativo. O HLA-G é bastante conhecido por estar envolvido na indução e na manutenção da tolerância entre o sistema imunológico materno e o feto semialogênico ao nível da interface fetoplacentária. Além disso, diversos estudos apontam para um papel imunorregulatório mais amplo dessa molécula. Neste contexto, a expressão de HLA-G em doenças inflamatórias e reumatológicas é uma área relativamente recente de pesquisa. Os primeiros estudos descreveram a expressão de HLA-G em várias miopatias inflamatórias, dermatite atópica e psoríase cutânea. Com base nos achados de que o HLA-G poderia desviar respostas T helper para o tipo Th2, foi levantada a hipótese de que o HLA-G seria uma molécula protetora nas respostas inflamatórias. Neste artigo, revisamos os potenciais papéis da molécula HLA-G no sistema imunológico e em diversas doenças reumatológicas, tais como lúpus eritematoso sistêmico, artrite reumatoide, esclerose sistêmica e outrasHuman leukocyte antigen G (HLA-G is a non-classic class I major histocompatibility complex (MHC molecule characterized by low polymorphism in its coding region, a limited tissue distribution pattern in physiologic conditions, and expression through soluble isoforms and isoforms bound to surface membranes through alternative splicing. HLA-G is fairly known since it is involved in induction and maintenance of tolerance between the maternal immunologic system and the semi-allogeneic fetus at the level of the fetal-placental interface. Besides, several studies have indicated a wider immunoregulatory role of

  16. O SUBLIME NA MODERNIDADE

    Directory of Open Access Journals (Sweden)

    Martha de Almeida

    2009-03-01

    Full Text Available A obra Uma investigação filosófica sobre a origem de nossas idéias do sublime e dobelo, do sensualista inglês Edmund Burke (1757, data as origens do sublime na Modernidade. Nosublime ocorre é um prazer ligado à dor, um "horror delicioso" que sentimos quando acreditamosque estamos em perigo sem que isso esteja ocorrendo realmente. Em O mundo como Vontade erepresentação Schopenhauer assume as influências que sofreu de Kant no que diz respeito à suainterpretação sobre o belo e o sublime, porém difere dele quanto à natureza dessa impressão. ParaSchopenhauer a experiência estética pressupõe a dissolução da subjetividade num movimento decontemplação das idéias livre do querer imposto pela vontade individual. As idéias deSchopenhauer influenciaram a estética do Nietzsche de O nascimento da tragédia.

  17. A aposta na filosofia

    Directory of Open Access Journals (Sweden)

    Márcio Suzuki

    2011-12-01

    Full Text Available Para Hume, filosofar é uma atividade semelhante às demais ocupações humanas. A decisão que leva à atividade filosófica é menos resultado de uma argumentação teórica do que fruto de um cálculo prático, que é entendido como uma caça ou um jogo. O objetivo deste trabalho será o de mostrar que no jogo filosófico perde quem joga sério demais. A aposta na filosofia tem que passar pelo bom humor e pela diversão: uma resposta a Pascal?For Hume, philosophy is in itself neither more nor less important than other human activities. On the contrary, it can be explained from its resemblance with game and hunting. If one chooses to dedicate himself to philosophy, his decision is not the consequence of logical reasoning, but depends on practical evaluation, which has something to do with gambling. The following text tries to show that in philosophy, like in other games, the loser will be the one who plays too earnestly. The philosophical game requires good humour and some relaxation: Is this a good answer to Pascal's pari?

  18. Gene Locater

    DEFF Research Database (Denmark)

    Anwar, Muhammad Zohaib; Sehar, Anoosha; Rehman, Inayat-Ur

    2012-01-01

    UNLABELLED: Locating genes on a chromosome is important for understanding the gene function and its linkage and recombination. Knowledge of gene positions on chromosomes is necessary for annotation. The study is essential for disease genetics and genomics, among other aspects. Currently available...... software's for calculating recombination frequency is mostly limited to the range and flexibility of this type of analysis. GENE LOCATER is a fully customizable program for calculating recombination frequency, written in JAVA. Through an easy-to-use interface, GENE LOCATOR allows users a high degree...

  19. Relationship between intracellular Na+ concentration and reduced Na+ affinity in Na+,K+-ATPase mutants causing neurological disease

    DEFF Research Database (Denmark)

    Toustrup-Jensen, Mads Schak; Einholm, Anja P.; Schack, Vivien

    2014-01-01

    The neurological disorders familial hemiplegic migraine type 2 (FHM2), alternating hemiplegia of childhood (AHC), and rapid-onset dystonia parkinsonism (RDP) are caused by mutations of Na+,K+-ATPase α2- and α3-isoforms, expressed in glial and neuronal cells, respectively. Although these disorders...... mutations that increase Na+ affinity were found to reduce [Na+]i. It is concluded that the Na+ affinity of the Na+,K+-ATPase is an important determinant of [Na+]i....

  20. Effect of salt stress on growth, Na+ accumulation and proline metabolism in potato (Solanum tuberosum cultivars.

    Directory of Open Access Journals (Sweden)

    Rinse Jaarsma

    Full Text Available Potato (Solanum tuberosum is a major crop world-wide and the productivity of currently used cultivars is strongly reduced at high soil salt levels. We compared the response of six potato cultivars to increased root NaCl concentrations. Cuttings were grown hydroponically and treated with 0 mM, 60 mM and 180 mM NaCl for one week. Growth reduction on salt was strongest for the cultivars Mozart and Mona Lisa with a severe senescence response at 180 mM NaCl and Mozart barely survived the treatment. The cultivars Desiree and Russett Burbank were more tolerant showing no senescence after salt treatment. A clear difference in Na(+ homeostasis was observed between sensitive and tolerant cultivars. The salt sensitive cultivar Mozart combined low Na(+ levels in root and stem with the highest leaf Na(+ concentration of all cultivars, resulting in a high Na(+ shoot distribution index (SDI for Mozart as compared to Desiree. Overall, a positive correlation between salt tolerance and stem Na(+ accumulation was found and the SDI for Na(+ points to a role of stem Na(+ accumulation in tolerance. In stem tissue, Mozart accumulated more H2O2 and less proline compared to the tolerant cultivars. Analysis of the expression of proline biosynthesis genes in Mozart and Desiree showed a clear reduction in proline dehydrogenase (PDH expression in both cultivars and an increase in pyrroline-5-carboxylate synthetase 1 (P5CS1 gene expression in Desiree, but not in Mozart. Taken together, current day commercial cultivars show promising differences in salt tolerance and the results suggest that mechanisms of tolerance reside in the capacity of Na(+ accumulation in stem tissue, resulting in reduced Na(+ transport to the leaves.

  1. Effect of Salt Stress on Growth, Na+ Accumulation and Proline Metabolism in Potato (Solanum tuberosum) Cultivars

    Science.gov (United States)

    Jaarsma, Rinse; de Vries, Rozemarijn S. M.; de Boer, Albertus H.

    2013-01-01

    Potato (Solanum tuberosum) is a major crop world-wide and the productivity of currently used cultivars is strongly reduced at high soil salt levels. We compared the response of six potato cultivars to increased root NaCl concentrations. Cuttings were grown hydroponically and treated with 0 mM, 60 mM and 180 mM NaCl for one week. Growth reduction on salt was strongest for the cultivars Mozart and Mona Lisa with a severe senescence response at 180 mM NaCl and Mozart barely survived the treatment. The cultivars Desiree and Russett Burbank were more tolerant showing no senescence after salt treatment. A clear difference in Na+ homeostasis was observed between sensitive and tolerant cultivars. The salt sensitive cultivar Mozart combined low Na+ levels in root and stem with the highest leaf Na+ concentration of all cultivars, resulting in a high Na+ shoot distribution index (SDI) for Mozart as compared to Desiree. Overall, a positive correlation between salt tolerance and stem Na+ accumulation was found and the SDI for Na+ points to a role of stem Na+ accumulation in tolerance. In stem tissue, Mozart accumulated more H2O2 and less proline compared to the tolerant cultivars. Analysis of the expression of proline biosynthesis genes in Mozart and Desiree showed a clear reduction in proline dehydrogenase (PDH) expression in both cultivars and an increase in pyrroline-5-carboxylate synthetase 1 (P5CS1) gene expression in Desiree, but not in Mozart. Taken together, current day commercial cultivars show promising differences in salt tolerance and the results suggest that mechanisms of tolerance reside in the capacity of Na+ accumulation in stem tissue, resulting in reduced Na+ transport to the leaves. PMID:23533673

  2. Effect of salt stress on growth, Na+ accumulation and proline metabolism in potato (Solanum tuberosum) cultivars.

    Science.gov (United States)

    Jaarsma, Rinse; de Vries, Rozemarijn S M; de Boer, Albertus H

    2013-01-01

    Potato (Solanum tuberosum) is a major crop world-wide and the productivity of currently used cultivars is strongly reduced at high soil salt levels. We compared the response of six potato cultivars to increased root NaCl concentrations. Cuttings were grown hydroponically and treated with 0 mM, 60 mM and 180 mM NaCl for one week. Growth reduction on salt was strongest for the cultivars Mozart and Mona Lisa with a severe senescence response at 180 mM NaCl and Mozart barely survived the treatment. The cultivars Desiree and Russett Burbank were more tolerant showing no senescence after salt treatment. A clear difference in Na(+) homeostasis was observed between sensitive and tolerant cultivars. The salt sensitive cultivar Mozart combined low Na(+) levels in root and stem with the highest leaf Na(+) concentration of all cultivars, resulting in a high Na(+) shoot distribution index (SDI) for Mozart as compared to Desiree. Overall, a positive correlation between salt tolerance and stem Na(+) accumulation was found and the SDI for Na(+) points to a role of stem Na(+) accumulation in tolerance. In stem tissue, Mozart accumulated more H2O2 and less proline compared to the tolerant cultivars. Analysis of the expression of proline biosynthesis genes in Mozart and Desiree showed a clear reduction in proline dehydrogenase (PDH) expression in both cultivars and an increase in pyrroline-5-carboxylate synthetase 1 (P5CS1) gene expression in Desiree, but not in Mozart. Taken together, current day commercial cultivars show promising differences in salt tolerance and the results suggest that mechanisms of tolerance reside in the capacity of Na(+) accumulation in stem tissue, resulting in reduced Na(+) transport to the leaves.

  3. Hemisferectomia na hemiplegia infantil

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    Rolando A. Tenuto

    1956-03-01

    Full Text Available Os autores relatam o caso de um paciente com 18 anos de idade, portador de paralisia cerebral instalada aos 4 anos de idade após moléstia febril prolongada (hemiplegia esquerda, crises convulsivas generalizadas, freqüentes e rebeldes à medicação, deficiência mental e alterações de conduta. O eletrencefalograma e o pneumencefalograma revelaram alterações graves do hemisfério direito; tendo o primeiro destes exames demonstrado a existência de perturbações propagadas ao outro hemisfério. Foi praticada a hemisferectomia direita. Durante o ato cirúrgico, por exigência técnica foram extirpados, além do hemisfério cerebral, os dois terços rostrais do núcleo caudado e o núcleo amigdalóide. No pós-operatório foi intercorrência de osteomielite e meningite, medicadas com antibióticos e seqüestrectomia. Apesar da complicação pós-operatória, instalou-se progressiva melhora em relação à conduta social; diminuiu também a espasticidade no membro inferior esquerdo e não mais se repetiram as crises convulsivas. Não foram observadas modificações quanto ao rendimento intelectual e quanto aos distúrbios sensitivos que existiam antes da intervenção cirúrgica. O exame neurocular mostrou hemianopsia homônima esquerda. Êstes resultados são concordantes com os assinalados na literatura sobre o assunto.

  4. TRGOVANJE NA IZBRANEM TRGU - FOREX

    OpenAIRE

    Ambrož, Sabina

    2012-01-01

    V delu diplomskega seminarja je obravnavana tematika trgovanja na valutnem trgu. Trgovanje z valutami je v zadnjem desetletju doseglo velik razvoj, močno pa je narasla tudi priljubljenost valutnega trga Forex, saj se zaradi enostavnega pristopa in možnosti velikih zaslužkov zanj odloča vse več posameznikov. V teoretičnem delu povzamemo ključne elemente, ki jih mora poznati začetnik na tem trgu. V empiričnem delu pa poskušamo na praktičnem primeru pokazati primer trgovanja s trendom in vstopom...

  5. [Na/K pump and intracellular monovalent cations: novel mechanism of excitation-transcription coupling involved in inhibition of apoptosis].

    Science.gov (United States)

    Taurin, S; Hamet, P; Orlov, S N

    2003-01-01

    The Na/K pump plays a key role in the regulation of the intracellular concentrations of monovalent cations and related cell function leading to electrogenesis and excitation-contraction coupling. We focus this review on the analysis of recent data showing that (i) inhibition of the Na/K pump triggers a signaling cascade independently of modulation of the intracellular [Na+]i/[K+]i ratio; (ii) elevation of [Na+]i under sustained inhibition of the Na/K pump leads to expression of a set of genes by [Ca2+]i-dependent and independent pathways; (iii) [Na+]i-sensitive genes are involved in the inhibition of programmed cell death (apoptosis) in vascular smooth muscle cells.

  6. The second sodium pump: from the function to the gene.

    Science.gov (United States)

    Rocafull, Miguel A; Thomas, Luz E; del Castillo, Jesús R

    2012-06-01

    Transepithelial Na(+) transport is mediated by passive Na(+) entry across the luminal membrane and exit through the basolateral membrane by two active mechanisms: the Na(+)/K(+) pump and the second sodium pump. These processes are associated with the ouabain-sensitive Na(+)/K(+)-ATPase and the ouabain-insensitive, furosemide-inhibitable Na(+)-ATPase, respectively. Over the last 40 years, the second sodium pump has not been successfully associated with any particular membrane protein. Recently, however, purification and cloning of intestinal α-subunit of the Na(+)-ATPase from guinea pig allowed us to define it as a unique biochemical and molecular entity. The Na(+)- and Na(+)/K(+)-ATPase genes are at the same locus, atp1a1, but have independent promoters and some different exons. Herein, we spotlight the functional characteristics of the second sodium pump, and the associated Na(+)-ATPase, in the context of its role in transepithelial transport and its response to a variety of physiological and pathophysiological conditions. Identification of the Na(+)-ATPase gene (atna) allowed us, using a bioinformatics approach, to explore the tertiary structure of the protein in relation to other P-type ATPases and to predict regulatory sites in the promoter region. Potential regulatory sites linked to inflammation and cellular stress were identified in the atna gene. In addition, a human atna ortholog was recognized. Finally, experimental data obtained using spontaneously hypertensive rats suggest that the Na(+)-ATPase could play a role in the pathogenesis of essential hypertension. Thus, the participation of the second sodium pump in transepithelial Na(+) transport and cellular Na(+) homeostasis leads us to reconsider its role in health and disease.

  7. Tuning of influenza A virus neuraminidase activity

    NARCIS (Netherlands)

    Dai, Meiling

    2017-01-01

    Influenza A viruses (IAVs) are zoonotic pathogens that constantly circulate in a wide variety of species, including birds, pigs and humans. In humans, IAVs cause seasonal epidemics and occasional influenza pandemics. Annual epidemics caused by seasonal IAVs usually lead to millions of human

  8. [From gene to disease; primary erythermalgia--a neuropathic disease as a consequence of mutations in a sodium pump gene

    NARCIS (Netherlands)

    Drenth, J.P.H.; Morsche, R.H.M. te; Michiels, J.J.M.

    2006-01-01

    Primary erythermalgia is a rare autosomal dominant inherited disorder characterized by recurrent attacks of red, warm and painful burning extremities. The gene involved in primary erythermalgia, SCN9A, encodes for a voltage dependent sodium channel alpha subunit (NaV1.7). NaV1.7 is located in dorsal

  9. Results from NA61/SHINE

    Directory of Open Access Journals (Sweden)

    Unger M.

    2013-06-01

    Full Text Available In this paper we summarize recent results from NA61/SHINE relevant for heavy ion physics, neutrino oscillations and the interpretation of air showers induced by ultra-high energy cosmic rays.

  10. ŠOLANJE NA DOMU

    OpenAIRE

    Horvat, Anja

    2016-01-01

    Pravica do izobraževanja in pridobivanja (ne)formalnih veščin in spretnosti je temeljna človekova pravica in svoboščina. Dobra izobrazba človeka duhovno bogati, plemeniti in širi njegova obzorja. Šolanje oz. izobraževanje na domu je ena od možnosti za izvajanje izobraževanja na osnovnošolski ravni, ki ga v Republiki Sloveniji ureja Zakon o osnovni šoli (1996, 5. in 88. člen). Ta predpisuje obvezne učne vsebine in načine preverjanja znanja, od izvajalcev poučevanja na domu pa ne zahteva do...

  11. Beam monitoring at NA2

    CERN Multimedia

    1978-01-01

    Claus Goessling working on the beam Cerenkov counter of NA2. The muon beam enters from left the hall EHN2 and the last element of the beam transport. On background is the access door on the Jura side.

  12. Na via do Behemoth

    Directory of Open Access Journals (Sweden)

    Gérard Rabinovitch

    2008-06-01

    Full Text Available O momento nazista permanece um enigma impensado de que a cultura contemporânea continua cativa. O autor levanta a hipótese de que a persistência desse enigma e seus estragos duráveis se devem à insistência dos pensamentos político, sociológico e filosófico em construírem o nazismo à luz da metáfora hobbesiana do Leviatã. Propõe retomar a questão do nazismo sob a perspectiva do Behemoth, antônimo do Leviatã. Para tanto, sugere a necessidade de retornar a Freud e à psicanálise, balizas de um possível novo pensamento do político que escrutine a destrutividade nazista. Propõe ainda sondar as homologias entre nazismo, corjas e máfias, com base na figura da " heroicização da violência" que lhes seria comum. E avança um modelo " econômico" : a quimera, suscetível de capturar o caráter heterotópico e heterocrônico de sua construção criminosa.The paths of Behemoth. The Nazi moment remains as the thoughtless enigma of which contemporary culture is still captive. The author raises the hypothesis where the persistency of this enigma and its durable damages are due to the insistence of the political, social and philosophical thoughts in building the Nazism at the view of the Hobbesian metaphor of Leviathan. It is here proposed to resume the Nazism matter under Behemoth's perspective, antonym to Leviathan. For such, it is suggested a necessity to look back at Freud and the psychoanalysis, structured by a possible new political thought which scrutinizes the destructivity of the Nazism. It is also here proposed to gaze at the homologies between Nazism, mafias and gangs, with a base in the figure of " violence as a heroically act" that are their common ground. The author also advances an economical model: the chimera susceptible to capture the heterotopic and heterochronic character of its criminal construction.

  13. Genetics of Na+ exclusion and salinity tolerance in Afghani durum wheat landraces

    KAUST Repository

    Shamaya, Nawar Jalal

    2017-11-21

    BackgroundSelecting for low concentration of Na+ in the shoot provides one approach for tackling salinity stress that adversely affects crop production. Novel alleles for Na+ exclusion can be identified and then introduced into elite crop cultivars.ResultsWe have identified loci associated with lower Na+ concentration in leaves of durum wheat landraces originating from Afghanistan. Seedlings of two F2 populations derived from crossings between Australian durum wheat (Jandaroi) and two Afghani landraces (AUS-14740 and AUS-14752) were grown hydroponically and evaluated for Na+ and K+ concentration in the third leaf. High heritability was found for both third leaf Na+ concentration and the K+/Na+ ratio in both populations. Further work focussed on line AUS-14740. Bulk segregant analysis using 9 K SNP markers identified two loci significantly associated with third leaf Na+ concentration. Marker regression analysis showed a strong association between all traits studied and a favourable allele originating from AUS-14740 located on the long arm of chromosome 4B.ConclusionsThe candidate gene in the relevant region of chromosome 4B is likely to be the high affinity K+ transporter B1 (HKT1;5-B1). A second locus associated with third leaf Na+ concentration was located on chromosome 3BL, with the favourable allele originating from Jandaroi; however, no candidate gene can be identified.

  14. Impact of childhood stress on psychopathology Impacto de estresse na infância na psicopatologia

    Directory of Open Access Journals (Sweden)

    Elisa Brietzke

    2012-12-01

    Full Text Available OBJECTIVE: Advances in our knowledge of mental disorder (MD genetics have contributed to a better understanding of their pathophysiology. Nonetheless, several questions and doubts persist. Recent studies have focused on environmental influences in the development of MDs, and the advent of neuroscientific methodologies has provided new perspectives. Early life events, such as childhood stress, may affect neurodevelopment through mechanisms such as gene-environment interactions and epigenetic regulation, thus leading to diseases in adulthood. The aim of this paper is to review the evidence regarding the role of the environment, particularly childhood stress, in the pathophysiology of MD. METHODOLOGY: We reviewed articles that evaluated environmental influences, with a particular focus on childhood trauma, brain morphology, cognitive functions, and the development of psychopathology and MD. RESULTS AND CONCLUSION: MRI studies have shown that exposure to trauma at an early age can result in several neurostructural changes, such as the reduction of the hippocampus and corpus callosum. Cognitive performance and functioning are also altered in this population. Finally, childhood stress is related to an increased risk of developing MD such as depression, bipolar disorder, schizophrenia and substance abuse. We conclude that there is robust evidence of the role of the environment, specifically adverse childhood experiences, in various aspects of MD.OBJETIVO: Avanços no conhecimento da genética dos transtornos mentais (TM contribuíram para um melhor entendimento de suas bases fisiopatológicas. No entanto, dúvidas e questões ainda persistem. Estudos recentes têm se concentrado nas influências do ambiente no desenvolvimento de TM, e o advento de metodologias neurocientíficas oferece novas perspectivas. Eventos precoces de vida, como estresse na infância, podem ser capazes de alterar o neurodesenvolvimento através de mecanismos como intera

  15. α3Na+/K+-ATPase deficiency causes brain ventricle dilation and abrupt embryonic motility in zebrafish

    DEFF Research Database (Denmark)

    Doganli, Canan; Beck, Hans Christian; Ribera, Angeles B

    2013-01-01

    Na+/K+-ATPases are transmembrane ion pumps that maintain ion gradients across the basolateral plasma membrane in all animal cells to facilitate essential biological functions. Mutations in the Na+/K+-ATPase α3 subunit gene (ATP1A3) cause rapid-onset dystonia-parkinsonism, a rare movement disorder...

  16. Na+/myo-inositol symporters and Na+/H+-antiport in Mesembryanthemum crystallinum.

    Science.gov (United States)

    Chauhan, S; Forsthoefel, N; Ran, Y; Quigley, F; Nelson, D E; Bohnert, H J

    2000-11-01

    Mitr1 and Mitr2 from Mesembryanthemum crystallinum (common ice plant) are members of a family of genes homologous to H+[or Na+]/myo-inositol symporters (ITRs), not previously studied in plants. MITR1 complemented an Itr1-deficient yeast strain. Mitr1 is strongly expressed in roots, moderately in stems, and weakly in leaves. Its transcripts increased in all organs, most dramatically in roots, under salinity stress. Mitr2 constitutes a rare transcript, slightly upregulated by salt stress in leaves only. Mitr1 transcripts are present in all cells in the root tip, but become restricted to phloem-associated cells in mature roots. Peptide antibodies against the two proteins indicated the presence of MITR1 in all organs and of MITR2 in leaves. Both are located in the tonoplast. MITR1 acts in removing sodium from root vacuoles, correlated with findings of low root sodium, while leaf vacuoles accumulate sodium in the ice plant. Up-regulation in leaves and stems is also found for Na+/H+-antiporter (Nhx-type) transcripts. Under comparable stress conditions, Nhx-and Itr-like transcripts in Arabidopsis were regulated differently. In the ice plant, co-ordinate induction of Na+/H+-antiporters and Na+/myo-inositol symporters transfers sodium from vacuoles in root cells into the leaf mesophyll as a halophytic strategy that lowers the osmotic potential. The tissue-specific differential expression of Itr- and Nhx-type transcripts suggests that the vacuolar sodium/inositol symporters function to reduce sodium amounts in cells of the root and vascular tissue, while sodium/proton antiporters in leaf tissues function to partition sodium into vacuoles for storage.

  17. Trichoderma genes

    Science.gov (United States)

    Foreman, Pamela [Los Altos, CA; Goedegebuur, Frits [Vlaardingen, NL; Van Solingen, Pieter [Naaldwijk, NL; Ward, Michael [San Francisco, CA

    2012-06-19

    Described herein are novel gene sequences isolated from Trichoderma reesei. Two genes encoding proteins comprising a cellulose binding domain, one encoding an arabionfuranosidase and one encoding an acetylxylanesterase are described. The sequences, CIP1 and CIP2, contain a cellulose binding domain. These proteins are especially useful in the textile and detergent industry and in pulp and paper industry.

  18. Alternative splice variant of the thiazide-sensitive NaCl cotransporter

    DEFF Research Database (Denmark)

    Tutakhel, Omar A Z; Jeleń, Sabina; Valdez-Flores, Marco

    2016-01-01

    The thiazide-sensitive NaCl cotransporter (NCC) is an important pharmacological target in the treatment of hypertension. Human SLC12A3 gene, encoding NCC, gives rise to three isoforms. Only the 3(rd) isoform has been extensively investigated. The aim of the present study was, therefore, to establ......The thiazide-sensitive NaCl cotransporter (NCC) is an important pharmacological target in the treatment of hypertension. Human SLC12A3 gene, encoding NCC, gives rise to three isoforms. Only the 3(rd) isoform has been extensively investigated. The aim of the present study was, therefore...... subjects significantly reduces the abundance of NCC1/2 in urinary exosomes. The present study highlights that previously underrepresented NCC1/2 is a fully functional thiazide-sensitive NaCl-transporting protein. Being significantly expressed in the kidney it may constitute a unique route of renal NaCl...

  19. Eletrocauterio na cesarea : complicações na ferida cirurgica

    OpenAIRE

    Cristiane Menabo Moreira

    2010-01-01

    Resumo: Objetivo: Avaliar a efetividade e segurança do uso do eletrocautério para coagulação na da ferida operatória da cesárea. Métodos: Foi realizado um ensaio clínico piloto duplo cego controlado aleatorizado. Mulheres com indicação de cesárea, com até uma cesárea prévia, que fizeram pré-natal foram aleatorizadas na hora da cirurgia para uso ou não de eletrocautério para coagulação. As 224 voluntárias que participaram foram examinadas na alta hospitalar (3o dia) e pós-alta, no 7°.ao 10° di...

  20. NqrM (DUF539) Protein Is Required for Maturation of Bacterial Na+-Translocating NADH:Quinone Oxidoreductase.

    Science.gov (United States)

    Kostyrko, Vitaly A; Bertsova, Yulia V; Serebryakova, Marina V; Baykov, Alexander A; Bogachev, Alexander V

    2015-12-07

    Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR) catalyzes electron transfer from NADH to ubiquinone in the bacterial respiratory chain, coupled with Na(+) translocation across the membrane. Na(+)-NQR maturation involves covalent attachment of flavin mononucleotide (FMN) residues, catalyzed by flavin transferase encoded by the nqr-associated apbE gene. Analysis of complete bacterial genomes has revealed another putative gene (duf539, here renamed nqrM) that usually follows the apbE gene and is present only in Na(+)-NQR-containing bacteria. Expression of the Vibrio harveyi nqr operon alone or with the associated apbE gene in Escherichia coli, which lacks its own Na(+)-NQR, resulted in an enzyme incapable of Na(+)-dependent NADH or reduced nicotinamide hypoxanthine dinucleotide (dNADH) oxidation. However, fully functional Na(+)-NQR was restored when these genes were coexpressed with the V. harveyi nqrM gene. Furthermore, nqrM lesions in Klebsiella pneumoniae and V. harveyi prevented production of functional Na(+)-NQR, which could be recovered by an nqrM-containing plasmid. The Na(+)-NQR complex isolated from the nqrM-deficient strain of V. harveyi lacks several subunits, indicating that nqrM is necessary for Na(+)-NQR assembly. The protein product of the nqrM gene, NqrM, contains a single putative transmembrane α-helix and four conserved Cys residues. Mutating one of these residues (Cys33 in V. harveyi NqrM) to Ser completely prevented Na(+)-NQR maturation, whereas mutating any other Cys residue only decreased the yield of the mature protein. These findings identify NqrM as the second specific maturation factor of Na(+)-NQR in proteobacteria, which is presumably involved in the delivery of Fe to form the (Cys)4[Fe] center between subunits NqrD and NqrE. Na(+)-translocating NADH:quinone oxidoreductase complex (Na(+)-NQR) is a unique primary Na(+) pump believed to enhance the vitality of many bacteria, including important pathogens such as Vibrio cholerae

  1. ChPT tests at NA48 and NA62

    Directory of Open Access Journals (Sweden)

    Gonnella Francesco

    2014-01-01

    Full Text Available Final results from an analysis of about 400 K± → π±γγ rare decay candidates collected by the NA48/2 and NA62 experiments at CERN during low intensity runs with minimum bias trigger configurations are presented. The results include a model-independent decay rate measurement and fits to Chiral Perturbation Theory (ChPT description. The data support the ChPT prediction for a cusp in the di-photon invariant mass spectrum at the two pion threshold.

  2. Corpo e sexualidade na gravidez

    Directory of Open Access Journals (Sweden)

    Natalúcia Matos Araújo

    2012-06-01

    Full Text Available Estudo etnográfico que teve como objetivo compreender como as gestantes vivenciam os processos fisiológicos do seu corpo durante a gestação e a sua repercussão na sexualidade. A pesquisa envolveu sete mulheres residentes em bairro popular de São Paulo. Na coleta de dados, utilizou-se observação participante e entrevista com questões norteadoras. Os dados foram apresentados na forma de narrativa e posteriormente organizados nas categorias: Percebendo as transformações corporais; Convivendo com as mudanças no corpo; Sentimentos e sensações na vida sexual durante a gestação e imaginando o corpo e a sexualidade após a gestação. As mulheres referiram-se às transformações do corpo como desconfortos e expressaram a preocupação de que fossem definitivas. Expressaram o desejo de que, após o parto, o corpo volte a ser como era e que volte a sentir desejo sexual. O reconhecimento destes fatos constitui-se numa ferramenta primordial na adequação das práticas profissionais.

  3. All new for NA62

    CERN Multimedia

    Antonella Del Rosso

    2012-01-01

    This week sees the start of the first run of the new NA62 experiment. This will be a unique opportunity for the collaboration to test its new beam, new detectors and new data acquisition system before the physics run in 2014. Speaking to the Bulletin, the NA62 technical coordinator Ferdinand Hahn shares the many challenges that the various teams faced to be on time for beam. Ready, steady, start!   A Large Angle Veto detector (white) in place in the NA62 decay volume (blue). With components from almost all the detectors in place downstream of the decay point of the mother particles – the kaons – and of the KTAG detector that tags the kaons before they decay, NA62 is ready for its first technical run. This unique run will test all the equipment as well as the trigger and the data acquisition systems. “This year, we will have about five weeks of beam from the SPS before the long shutdown of all the CERN machines,” says Ferdinand Hahn, NA62 Technical Co-...

  4. Antisense-mediated knockdown of Na(V)1.8, but not Na(V)1.9, generates inhibitory effects on complete Freund's adjuvant-induced inflammatory pain in rat.

    Science.gov (United States)

    Yu, Yao-Qing; Zhao, Feng; Guan, Su-Min; Chen, Jun

    2011-05-10

    Tetrodotoxin-resistant (TTX-R) sodium channels Na(V)1.8 and Na(V)1.9 in sensory neurons were known as key pain modulators. Comparing with the widely reported Na(V)1.8, roles of Na(V)1.9 on inflammatory pain are poorly studied by antisense-induced specific gene knockdown. Here, we used molecular, electrophysiological and behavioral methods to examine the effects of antisense oligodeoxynucleotide (AS ODN) targeting Na(V)1.8 and Na(V)1.9 on inflammatory pain. Following complete Freund's adjuvant (CFA) inflammation treatment, Na(V)1.8 and Na(V)1.9 in rat dorsal root ganglion (DRG) up-regulated mRNA and protein expressions and increased sodium current densities. Immunohistochemical data demonstrated that Na(V)1.8 mainly localized in medium and small-sized DRG neurons, whereas Na(V)1.9 only expressed in small-sized DRG neurons. Intrathecal (i.t.) delivery of AS ODN was used to down-regulate Na(V)1.8 or Na(V)1.9 expressions confirmed by immunohistochemistry and western blot. Unexpectedly, behavioral tests showed that only Na(V)1.8 AS ODN, but not Na(V)1.9 AS ODN could reverse CFA-induced heat and mechanical hypersensitivity. Our data indicated that TTX-R sodium channels Na(V)1.8 and Na(V)1.9 in primary sensory neurons played distinct roles in CFA-induced inflammatory pain and suggested that antisense oligodeoxynucleotide-mediated blocking of key pain modulator might point toward a potential treatment strategy against certain types of inflammatory pain.

  5. Antisense-Mediated Knockdown of NaV1.8, but Not NaV1.9, Generates Inhibitory Effects on Complete Freund's Adjuvant-Induced Inflammatory Pain in Rat

    Science.gov (United States)

    Guan, Su-Min; Chen, Jun

    2011-01-01

    Tetrodotoxin-resistant (TTX-R) sodium channels NaV1.8 and NaV1.9 in sensory neurons were known as key pain modulators. Comparing with the widely reported NaV1.8, roles of NaV1.9 on inflammatory pain are poorly studied by antisense-induced specific gene knockdown. Here, we used molecular, electrophysiological and behavioral methods to examine the effects of antisense oligodeoxynucleotide (AS ODN) targeting NaV1.8 and NaV1.9 on inflammatory pain. Following complete Freund's adjuvant (CFA) inflammation treatment, NaV1.8 and NaV1.9 in rat dorsal root ganglion (DRG) up-regulated mRNA and protein expressions and increased sodium current densities. Immunohistochemical data demonstrated that NaV1.8 mainly localized in medium and small-sized DRG neurons, whereas NaV1.9 only expressed in small-sized DRG neurons. Intrathecal (i.t.) delivery of AS ODN was used to down-regulate NaV1.8 or NaV1.9 expressions confirmed by immunohistochemistry and western blot. Unexpectedly, behavioral tests showed that only NaV1.8 AS ODN, but not NaV1.9 AS ODN could reverse CFA-induced heat and mechanical hypersensitivity. Our data indicated that TTX-R sodium channels NaV1.8 and NaV1.9 in primary sensory neurons played distinct roles in CFA-induced inflammatory pain and suggested that antisense oligodeoxynucleotide-mediated blocking of key pain modulator might point toward a potential treatment strategy against certain types of inflammatory pain. PMID:21572961

  6. A designated centre for people with disabilities operated by St Joseph's Foundation, Limerick

    LENUS (Irish Health Repository)

    Shi, Weifeng

    2010-12-01

    More and more nucleotide sequences of type A influenza virus are available in public databases. Although these sequences have been the focus of many molecular epidemiological and phylogenetic analyses, most studies only deal with a few representative sequences. In this paper, we present a complete analysis of all Haemagglutinin (HA) and Neuraminidase (NA) gene sequences available to allow large scale analyses of the evolution and epidemiology of type A influenza.

  7. A designated centre for people with disabilities operated by Ability West, Galway

    LENUS (Irish Health Repository)

    Shi, Weifeng

    2010-12-01

    More and more nucleotide sequences of type A influenza virus are available in public databases. Although these sequences have been the focus of many molecular epidemiological and phylogenetic analyses, most studies only deal with a few representative sequences. In this paper, we present a complete analysis of all Haemagglutinin (HA) and Neuraminidase (NA) gene sequences available to allow large scale analyses of the evolution and epidemiology of type A influenza.

  8. A designated centre for people with disabilities operated by St Christopher's Services Limited, Longford

    LENUS (Irish Health Repository)

    Shi, Weifeng

    2010-12-01

    More and more nucleotide sequences of type A influenza virus are available in public databases. Although these sequences have been the focus of many molecular epidemiological and phylogenetic analyses, most studies only deal with a few representative sequences. In this paper, we present a complete analysis of all Haemagglutinin (HA) and Neuraminidase (NA) gene sequences available to allow large scale analyses of the evolution and epidemiology of type A influenza.

  9. St Ita's Community Hospital, Newcastlewest, Limerick.

    LENUS (Irish Health Repository)

    Shi, Weifeng

    2010-12-01

    More and more nucleotide sequences of type A influenza virus are available in public databases. Although these sequences have been the focus of many molecular epidemiological and phylogenetic analyses, most studies only deal with a few representative sequences. In this paper, we present a complete analysis of all Haemagglutinin (HA) and Neuraminidase (NA) gene sequences available to allow large scale analyses of the evolution and epidemiology of type A influenza.

  10. Correction of the Caulobacter crescentus NA1000 genome annotation.

    Directory of Open Access Journals (Sweden)

    Bert Ely

    Full Text Available Bacterial genome annotations are accumulating rapidly in the GenBank database and the use of automated annotation technologies to create these annotations has become the norm. However, these automated methods commonly result in a small, but significant percentage of genome annotation errors. To improve accuracy and reliability, we analyzed the Caulobacter crescentus NA1000 genome utilizing computer programs Artemis and MICheck to manually examine the third codon position GC content, alignment to a third codon position GC frame plot peak, and matches in the GenBank database. We identified 11 new genes, modified the start site of 113 genes, and changed the reading frame of 38 genes that had been incorrectly annotated. Furthermore, our manual method of identifying protein-coding genes allowed us to remove 112 non-coding regions that had been designated as coding regions. The improved NA1000 genome annotation resulted in a reduction in the use of rare codons since noncoding regions with atypical codon usage were removed from the annotation and 49 new coding regions were added to the annotation. Thus, a more accurate codon usage table was generated as well. These results demonstrate that a comparison of the location of peaks third codon position GC content to the location of protein coding regions could be used to verify the annotation of any genome that has a GC content that is greater than 60%.

  11. gigapanorama of NA 62 cavern

    CERN Multimedia

    Brice, Maximilien

    2015-01-01

    The image shows the new rare Kaon decay experiment at CERN, called NA62. The NA62 experiment is 270 metres long and includes a 120-metre-long vacuum tank, shown here, housing several of the particle detectors. (Note: the experiment axis is a straight line, the curving of the tank is an optical effect of the photo.) Kaons are particles that decay into lighter elementary particles. The kaon decay processes are mostly well known, except for some very rare decay modes. For example, NA62 is investigating a rare decay predicted by the Standard Model in which a kaon decays into one pion and two neutrinos. This process occurs only once every 10 billion decays. The understanding of such ultra-rare decays are of great importance because they test the Standard Model in energy ranges not accessible by direct measurements. They are therefore complementary to the measurements at the LHC. ultra high definition on demand (photolab@cern.ch).

  12. Alcool e drogas na esquizofrenia

    OpenAIRE

    Marilia Montoya Boscolo

    2000-01-01

    Resumo: O objetivo deste trabalho é estudar o fenômeno do uso de álcool e drogas ilícitas (maconha e cocaína) na esquizoftenia. Trata-se de estudo comparativo, casocontrole, de dois grupos de esquizoftênicos pareados para sexo e idade, que preencheram os critérios diagnósticos do DSMIV para esquizoftenia. O grupo caso é definido por esquizofrênicos que têm história de ter feito uso de álcool, pelo menos no último mês e/ ou história de ter feito uso de drogas (maconha e cocaína), pelo menos al...

  13. OGLAŠEVANJE NA FACEBOOKU

    OpenAIRE

    Renko, Katica

    2013-01-01

    V magistrskem delu smo se seznanili z oglaševanjem na Facebooku. S pregledom gradiva, ki nam je ponujeno, smo preverili pogoje, načine, oblike … oglaševanja, ki pa smo jih s pregledom situacije v praksi preverili in dopolnili. Ob zaključku dela lahko rečemo, da je oglaševanje preko FB enostavnejše, cenejše, dostopnejše, omogoča nam ažurnost in samo komunikacijo z naslovniki, ki so potencialni kupci naših ponudb. Množični mediji nam omogočajo mnogotero obliko oglaševanja. Izbrana oblika ponuja...

  14. Multiplex Evaluation of Influenza Neutralizing Antibodies with Potential Applicability to In-Field Serological Studies

    National Research Council Canada - National Science Library

    Molesti, Eleonora; Wright, Edward; Terregino, Calogero; Rahman, Rafat; Cattoli, Giovanni; Temperton, Nigel J

    2014-01-01

    .... Retroviral pseudotypes bearing influenza haemagglutinin (HA) and neuraminidase (NA) envelope glycoproteins represent a flexible platform for sensitive, readily standardized influenza serological assays...

  15. Control of Neurotransmission by NaV1.7 in Human, Guinea Pig, and Mouse Airway Parasympathetic Nerves.

    Science.gov (United States)

    Kocmalova, Michaela; Kollarik, Marian; Canning, Brendan J; Ru, Fei; Adam Herbstsomer, R; Meeker, Sonya; Fonquerna, Silvia; Aparici, Monica; Miralpeix, Montserrat; Chi, Xian Xuan; Li, Baolin; Wilenkin, Ben; McDermott, Jeff; Nisenbaum, Eric; Krajewski, Jeffrey L; Undem, Bradley J

    2017-04-01

    Little is known about the neuronal voltage-gated sodium channels (NaVs) that control neurotransmission in the parasympathetic nervous system. We evaluated the expression of the α subunits of each of the nine NaVs in human, guinea pig, and mouse airway parasympathetic ganglia. We combined this information with a pharmacological analysis of selective NaV blockers on parasympathetic contractions of isolated airway smooth muscle. As would be expected from previous studies, tetrodotoxin potently blocked the parasympathetic responses in the airways of each species. Gene expression analysis showed that that NaV 1.7 was virtually the only tetrodotoxin-sensitive NaV1 gene expressed in guinea pig and human airway parasympathetic ganglia, where mouse ganglia expressed NaV1.1, 1.3, and 1.7. Using selective pharmacological blockers supported the gene expression results, showing that blocking NaV1.7 alone can abolish the responses in guinea pig and human bronchi, but not in mouse airways. To block the responses in mouse airways requires that NaV1.7 along with NaV1.1 and/or NaV1.3 is blocked. These results may suggest novel indications for NaV1.7-blocking drugs, in which there is an overactive parasympathetic drive, such as in asthma. The data also raise the potential concern of antiparasympathetic side effects for systemic NaV1.7 blockers. Copyright © 2017 by The American Society for Pharmacology and Experimental Therapeutics.

  16. HERKUL NA BRAČU

    OpenAIRE

    Cambi, Nenad

    2013-01-01

    U radu se raspravlja o reljefu Herkula uklesanom u litici pred ulazom u kamenolomu Rasohe. Figure uklesane u stijeni su kultni reljefi božanstva koje se časti u prirodnom okruženju. Herkulov reljef je kultna slika na ulazu u kamenolom pred kojim su se prinosile žrtve na kamenom žrtveniku. Također se raspravlja o natpisu sa žrtvenika posvećenom Herkulu iz kamenoloma Stražišće i donosi ispravak pogrešnog čitanja natpisa. Donose se i drugi nalazi iz kamenoloma Dalmcije koji ...

  17. Transcriptional Response of Selenopolypeptide Genes and Selenocysteine Biosynthesis Machinery Genes in Escherichia coli during Selenite Reduction

    Directory of Open Access Journals (Sweden)

    Antonia Y. Tetteh

    2014-01-01

    Full Text Available Bacteria can reduce toxic selenite into less toxic, elemental selenium (Se0, but the mechanism on how bacterial cells reduce selenite at molecular level is still not clear. We used Escherichia coli strain K12, a common bacterial strain, as a model to study its growth response to sodium selenite (Na2SeO3 treatment and then used quantitative real-time PCR (qRT-PCR to quantify transcript levels of three E. coli selenopolypeptide genes and a set of machinery genes for selenocysteine (SeCys biosynthesis and incorporation into polypeptides, whose involvements in the selenite reduction are largely unknown. We determined that 5 mM Na2SeO3 treatment inhibited growth by ∼50% while 0.001 to 0.01 mM treatments stimulated cell growth by ∼30%. Under 50% inhibitory or 30% stimulatory Na2SeO3 concentration, selenopolypeptide genes (fdnG, fdoG, and fdhF whose products require SeCys but not SeCys biosynthesis machinery genes were found to be induced ≥2-fold. In addition, one sulfur (S metabolic gene iscS and two previously reported selenite-responsive genes sodA and gutS were also induced ≥2-fold under 50% inhibitory concentration. Our findings provide insight about the detoxification of selenite in E. coli via induction of these genes involved in the selenite reduction process.

  18. [Na+, K(+)-ATPase, endogenous cardiotonic steroids and their transducing role].

    Science.gov (United States)

    Tsymbaliuk, O V; Kosterin, S O

    2012-01-01

    Na+, K(+)-ATPase--a protein complex of plasmatic membrane, which performs the dual function: firstly, it supports the Na+ and K+ homeostasis, and also transmembrane potential gradient, secondly, it serves as the transducer of signals and as the regulator of the expression of many key genes. Endogenous cardiotonic steroids, which are synthesized in the adrenal glands and hypothalamus, serve as the signal molecules. New concepts about the mechanisms of the realization of the Na+, K(+)-ATPase signal function and their connection with cellular functions, apoptosis, and with pathologies of cardiovascular system and water-salt homeostasis are described in the survey.

  19. Advances in understanding the genetic basis of inherited single gene skin barrier disorders: new clues to key genes that may be involved in the pathogenesis of atopic dermatitis Avanços no entendimento da base genética de doenças hereditárias monogênicas da barreira epidérmica: novas pistas para os principais genes que podem estar envolvidos na patogênese da dermatite atópica

    Directory of Open Access Journals (Sweden)

    Joey E Lai-Cheong

    2006-12-01

    Full Text Available Increasing knowledge of genomic DNA sequences and genetic databases has led to the characterization of the molecular basis of several inherited skin disorders. In this review we summarize some of the major recent discoveries that have been made in defining the pathogenic mutations that cause inherited disorders of the skin barrier leading to skin scaling or increased transepidermal water loss in either rare disorders (Netherton’s syndrome or harlequin ichthyosis or more common genodermatoses (ichthyosis vulgaris. These molecular breakthroughs have led to more accurate diagnoses, better genetic counselling and, where appropriate, the feasibility of DNA-based prenatal diagnosis, as well as the possibility of developing newer forms of treatment, including gene or protein therapy. Identifying the molecular basis of these conditions, especially ichthyosis vulgaris, has also provided dramatic new insight into the genetic abnormalities in the common disorder, atopic dermatitis. Thus research on the relatively rare single gene inherited skin disorders not only has benefits for patients and their families with these uncommon conditions but also has the potential to yield fresh and significant new information about very common skin diseases.O maior conhecimento sobre as de sequências genômicas de DNA e as bases de dados genéticas levou à caracterização da base molecular de várias doenças hereditárias de pele. Nesta revisão resumimos algumas das descobertas recentes mais importantes quanto à definição das mutações patogênicas que causam as doenças hereditárias da barreira cutânea, levando a descamação ou aumento da perda hídrica transepidérmica, seja em doenças raras, (síndrome de Netherton ou ictiose em Arlequim ou genodermatoses mais comuns (ictiose vulgar. Estas descobertas moleculares têm conduzido a diagnósticos mais acurados, melhor aconselhamento genético e, quando apropriado, à possibilidade de diagnóstico pr

  20. Identification of GPD1 gene from yeast via fluorescence differential ...

    African Journals Online (AJOL)

    The main task of this work was to identify abiotic stress-induced gene(s) from yeast (Saccharomyces cerevisiae) and introduce it to a prokaryotic system to detect its effect on conferring tolerance to salt stress. Six isolates of yeast (S. cerevisiae) were evaluated under salt and osmotic stresses at concentrations of 2 M NaCl ...

  1. Gene effects for resistance to groundnut rossette disease in exotic ...

    African Journals Online (AJOL)

    The materials were evaluated for biotic and abiotic stresses, but succumbed to groundnut rosette disease (GRD). For these superior lines to find utility in Uganda, they need further improvement by introducing resistance genes to GRD. A study was conducted at NaSARRI to determine nature of gene action controlling ...

  2. Gene Therapy

    Science.gov (United States)

    ... or improve your body's ability to fight disease. Gene therapy holds promise for treating a wide range of diseases, such as cancer, cystic fibrosis, heart disease, diabetes, hemophilia and AIDS. Researchers are still studying how and ...

  3. NaCl stress impact on the key enzymes in glycolysis from Lactobacillus bulgaricus during freeze-drying.

    Science.gov (United States)

    Li, Chun; Sun, Jinwei; Qi, Xiaoxi; Liu, Libo

    2015-01-01

    The viability of Lactobacillus bulgaricus in freeze-drying is of significant commercial interest to dairy industries. In the study, L.bulgaricus demonstrated a significantly improved (p enzymes in glycolysis during 2% NaCl stress were studied. NaCl stress significantly enhanced (p enzymes (phosphofructokinase, pyruvate kinase, and lactate dehydrogenase) decreased during freeze-drying, and NaCl stress were found to improve activities of these enzymes before and after freeze-drying. However, a transcriptional analysis of the corresponding genes suggested that the effect of NaCl stress on the expression of the pfk2 gene was not obvious. The increased survival of freeze-dried cells of L. bulgaricus under NaCl stress might be due to changes in only the activity or translation level of these enzymes in different environmental conditions but have no relation to their mRNA transcription level.

  4. NA60 frees the quarks

    CERN Multimedia

    2003-01-01

    Fitted with new state-of-the-art silicon detectors, NA60 is prepared to study the phase transition from confined hadronic matter to a deconfined (free) quark-gluon plasma, a state of matter which probably existed an instant after the Big Bang.

  5. NA62: Hidden Sector Physics

    CERN Document Server

    Cesarotti, Carissa Joyce

    2016-01-01

    Modern experimental physics is often probing for new physics by either finding deviations from predictions on extremely precise measurements, or by looking for a new signal that cannot be explained with existing models. The NA62 experiment at CERN does the former by measuring the ultra-rare decay $K^+ \\rightarrow \\pi^+ \

  6. EMC anténa

    OpenAIRE

    Tenora, Jan

    2016-01-01

    Cílem této diplomové práce bylo navrhnout anténu pracující ve frekvenčním pásmu 30 MHz až 1 GHz. Navržená bikónická anténa vyžaduje ke správné funkci symetrické napájení, proto bylo dále nutné navrhnout vhodný symetrizační člen sestavený z diskrétních součástek. Navržená anténa byla také zkonstruována a měřením byl ověřen její činitel odrazu na vstupu, zisk a směrové charakteristiky. Tato diplomová práce dále obsahuje seznámení čtenáře s principy měření úrovně rušivých signálů v oblasti EMC p...

  7. Characterization of duplicated Dunaliella viridis SPT1 genes provides insights into early gene divergence after duplication.

    Science.gov (United States)

    Guan, Zhenwei; Meng, Xiangzong; Sun, Zhenhua; Xu, Zhengkai; Song, Rentao

    2008-10-15

    The sodium-dependent phosphate transporter gene from unicellular green algae Dunaliella viridis, DvSPT1, shares similarity with members of Pi transporter family. Sequencing analysis of D. viridis BAC clone containing the DvSPT1 gene revealed two inverted duplicated copies of this gene (DvSPT1 and DvSPT1-2 respectively). The duplication covered most of both genes except for their 3' downstream region. The duplicated genomic sequences exhibited 97.9% identity with a synonymous divergence of Ks=0.0126 in the coding region. This data indicated very recent gene duplication in D. viridis genome, providing an excellent opportunity to investigate sequence and expression divergence of duplicated genes at an early stage. Scattered point mutations and length polymorphism of simple sequence repeats (SSRs) were predominant among the sequence divergence soon after gene duplication. Due to sequence divergence in the 5' regulatory regions and a swap of the entire 3' downstream regions (3'-UTR), DvSPT1 and DvSPT1-2 showed expression divergence in response to extra-cellular NaCl concentration changes. According to their expression patterns, the two diverged gene copies would provide better adaptation to a broader range of extra-cellular NaCl concentration. Furthermore, Southern blot analysis indicated that there might be a large phosphate transporter gene family in D. viridis.

  8. Global transcriptome profiling of wild soybean (Glycine soja) roots under NaHCO3 treatment.

    Science.gov (United States)

    Ge, Ying; Li, Yong; Zhu, Yan-Ming; Bai, Xi; Lv, De-Kang; Guo, Dianjing; Ji, Wei; Cai, Hua

    2010-07-26

    Plant roots are the primary site of perception and injury for saline-alkaline stress. The current knowledge of saline-alkaline stress transcriptome is mostly focused on saline (NaCl) stress and only limited information on alkaline (NaHCO3) stress is available. Using Affymetrix Soybean GeneChip, we conducted transcriptional profiling on Glycine soja roots subjected to 50 mmol/L NaHCO3 treatment. In a total of 7088 probe sets, 3307 were up-regulated and 5720 were down-regulated at various time points. The number of significantly stress regulated genes increased dramatically after 3 h stress treatment and peaked at 6 h. GO enrichment test revealed that most of the differentially expressed genes were involved in signal transduction, energy, transcription, secondary metabolism, transporter, disease and defence response. We also detected 11 microRNAs regulated by NaHCO3 stress. This is the first comprehensive wild soybean root transcriptome analysis under alkaline stress. These analyses have identified an inventory of genes with altered expression regulated by alkaline stress. The data extend the current understanding of wild soybean alkali stress response by providing a set of robustly selected, differentially expressed genes for further investigation.

  9. The F Gene of Rodent Brain-Adapted Mumps Virus Is a Major Determinant of Neurovirulence▿

    Science.gov (United States)

    Lemon, Ken; Rima, Bertus K.; McQuaid, Stephen; Allen, Ingrid V.; Duprex, W. Paul

    2007-01-01

    Prior to the introduction of live-attenuated vaccines, mumps virus (MuV) was the leading cause of virus-induced meningitis. Although vaccination has been effective at controlling the disease, the use of insufficiently attenuated strains has been associated with high rates of aseptic meningitis in vaccinees. The molecular basis of MuV attenuation is poorly understood, and no reliable molecular markers of virulence have been identified. In this study, reverse genetics has been used to identify molecular determinants of MuV neuropathogenesis. Recombinant viruses, containing the envelope-associated genes from the Kilham (MuVKH) rodent brain-adapted strain of MuV, were generated in the Jeryl Lynn 5 (MuVJL5) vaccine strain background. The syncytium phenotypes of the recombinant viruses on Vero cells differed depending on the source of the fusion (F) and hemagglutinin-neuraminidase (HN) glycoproteins, with heterologous combinations showing either an increase or a decrease in the level of cell fusion compared to that of the homologous parental combinations. This was confirmed by transiently cotransfecting eukaryotic F and HN glycoprotein expression constructs. A Lewis rat model that discriminates between neurovirulent and nonneurovirulent MuV strains based on the extent of hydrocephalus induced in the rat brain after intracerebral inoculation was used to assess the phenotype of the recombinant viruses. Expression of the matrix (M), small hydrophobic (SH), or HN gene in isolation did not confer a neurovirulent phenotype. Expression of the F gene of the neurovirulent strain alone was sufficient to induce significant levels of hydrocephalus. Coexpression of the homologous HN gene led to a marginal increase in the level of hydrocephalus. PMID:17475640

  10. Photodesorption of Na atoms from rough Na surfaces

    DEFF Research Database (Denmark)

    Balzer, Frank; Gerlach, R.; Manson, J.R.

    1997-01-01

    We investigate the desorption of Na atoms from large Na clusters deposited on dielectric surfaces. High-resolution translational energy distributions of the desorbing atoms are determined by three independent methods, two-photon laser-induced fluorescence, as well as single-photon and resonance......-enhanced two-photon ionization techniques. Upon variation of surface temperature and for different substrates (mica vs lithium fluoride) clear non-Maxwellian time-of-flight distributions are observed with a cos θ angular dependence and most probable kinetic energies below that expected of atoms desorbing from...... atoms are scattered by surface vibrations. Recent experiments providing time constants for the decay of the optical excitations in the clusters support this model. The excellent agreement between experiment and theory indicates the importance of both absorption of the laser photons via direct excitation...

  11. Retinoschisin is linked to retinal Na/K-ATPase signaling and localization.

    Science.gov (United States)

    Plössl, Karolina; Royer, Melanie; Bernklau, Sarah; Tavraz, Neslihan N; Friedrich, Thomas; Wild, Jens; Weber, Bernhard H F; Friedrich, Ulrike

    2017-08-01

    Mutations in the RS1 gene cause X-linked juvenile retinoschisis (XLRS), a hereditary retinal dystrophy. We recently showed that retinoschisin, the protein encoded by RS1 , regulates ERK signaling and apoptosis in retinal cells. In this study, we explored an influence of retinoschisin on the functionality of the Na/K-ATPase, its interaction partner at retinal plasma membranes. We show that retinoschisin binding requires the β2-subunit of the Na/K-ATPase, whereas the α-subunit is exchangeable. Our investigations revealed no effect of retinoschisin on Na/K-ATPase-mediated ATP hydrolysis and ion transport. However, we identified an influence of retinoschisin on Na/K-ATPase-regulated signaling cascades and Na/K-ATPase localization. In addition to the known ERK deactivation, retinoschisin treatment of retinoschisin-deficient ( Rs1h -/Y ) murine retinal explants decreased activation of Src, an initial transmitter in Na/K-ATPase signal transduction, and of Ca 2+ signaling marker Camk2. Immunohistochemistry on murine retinae revealed an overlap of the retinoschisin-Na/K-ATPase complex with proteins involved in Na/K-ATPase signaling, such as caveolin, phospholipase C, Src, and the IP3 receptor. Finally, retinoschisin treatment altered Na/K-ATPase localization in photoreceptors of Rs1h -/Y retinae. Taken together, our results suggest a regulatory effect of retinoschisin on Na/K-ATPase signaling and localization, whereas Na/K-ATPase-dysregulation caused by retinoschisin deficiency could represent an initial step in XLRS pathogenesis. © 2017 Plössl et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  12. Economia na consulta de Endodontia

    OpenAIRE

    Pinto, Diogo Filipe Cardoso

    2016-01-01

    Introdução: Em Portugal a realidade actual, clínica e financeira do exercício da Medicina Dentária é bem distinta da do final do séc. XX devido à pletora de Médicos Dentistas e à baixa de honorários por acto Médico registada nos últimos anos. Objectivos: Este trabalho teve como objectivo perceber um pouco sobre os detalhes a ter em consideração aquando da abertura de uma clínica centrada na realização de tratamentos na área da Endodontia, e como publicitá-la de forma legal e apelativa. Pre...

  13. ULIČNA LUTKOVNA PREDSTAVA

    OpenAIRE

    Novak, Natalija

    2016-01-01

    V diplomski nalogi z naslovom Ulična lutkovna predstava je predstavljena priprava in izvedba gledališko-lutkovne predstave, ki smo jo izvedli v okviru programa Šole uličnega gledališča, pod vodstvom gledališča Ane Monro. V teoretičnem delu smo na podlagi strokovne literature predstavili dejavnost uličnega gledališča, njegove zvrsti, vlogo prostora in publike v uličnem gledališču ter kratko zgodovino le-tega. Predstavili smo tudi gledališče Ane Monro, ki je glavni organizator Šole uličnega ...

  14. Carne suína

    OpenAIRE

    Ortigara, Claudino

    2000-01-01

    Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico. A carne suína é a fonte de proteína animal mais consumida no mundo. No entanto, em muitos países e em especial no Brasil o consumo per capita é muito baixo. Como conseqüência disso, limita-se o crescimento do segmento produtivo e uma série de benefícios socioeconômicos que são gerados pela atividade suinocultura.Com esse trabalho, buscamos uma explicação para entender a resistência da população ao consum...

  15. NA64 Status Report 2017

    CERN Document Server

    Gninenko, Sergei

    2017-01-01

    The experiment NA64 is aimed at a direct search for invisible decays of sub-GeV dark photons (A′). The main goal in 2016 was to probe a region of the A′ parameter space, particularly interesting for the explanation of the muon g − 2 anomaly. The status and results from two NA64 runs in July and October 2016, obtained, respectively, with 2.75×109 and 4×1010 accumulated electron on target are reported. A feasibility study of the search for the X → e+e− decay of a new light X boson, which could explain a recently observed excess of e+e− events from excited 8Be transitions is also presented.

  16. Chaperone-mediated gene therapy with recombinant AAV-PPCA in a new mouse model of type I sialidosis.

    Science.gov (United States)

    Bonten, Erik J; Yogalingam, Gouri; Hu, Huimin; Gomero, Elida; van de Vlekkert, Diantha; d'Azzo, Alessandra

    2013-10-01

    The lysosomal storage disease sialidosis is caused by a primary deficiency of the sialidase N-acetyl-α-neuraminidase-1 (NEU1). Patients with type I sialidosis develop an attenuated, non-neuropathic form of the disease also named cherry red spot myoclonus syndrome, with symptoms arising during juvenile/ adult age. NEU1 requires binding to its chaperone, protective protein/cathepsin A (PPCA), for lysosomal compartmentalization, stability and catalytic activation. We have generated a new mouse model of type I sialidosis that ubiquitously expresses a NEU1 variant carrying a V54M amino acid substitution identified in an adult patient with type I sialidosis. Mutant mice developed signs of lysosomal disease after 1year of age, predominantly in the kidney, albeit low residual NEU1 activity was detected in most organs and cell types. We demonstrate that the activity of the mutant enzyme could be effectively increased in all systemic tissues by chaperone-mediated gene therapy with a liver-tropic recombinant AAV2/8 vector expressing PPCA. This resulted in clear amelioration of the disease phenotype. These results suggest that at least some of the NEU1 mutations associated with type I sialidosis may respond to PPCA-chaperone-mediated gene therapy. Published by Elsevier B.V.

  17. NA

    OpenAIRE

    Gleeson, David A.

    1998-01-01

    With advancements in the fields of propulsion, aerodynamics, structures, materials and controls, the routine exploration of hypersonic, atmospheric flight has become a more feasible concept. Thus, there is a need for efficient and effective hypersonic configurations. Current studies in configuration efficiency and effectiveness seem to be concentrated in aircraft subsystem design, especially propulsion systems, rather than at the conceptual aircraft system design level. This thesis attempts t...

  18. Na+/H+ antiporters are differentially regulated in response to NaCl stress in leaves and roots of Mesembryanthemum crystallinum.

    Science.gov (United States)

    Cosentino, Cristian; Fischer-Schliebs, Elke; Bertl, Adam; Thiel, Gerhard; Homann, Ulrike

    2010-05-01

    Salinity tolerance in plants involves controlled Na(+) transport at the site of Na(+) accumulation and intracellular Na(+) compartmentation. The focus of this study was the identification and analysis of the expression of Na(+)/H(+) antiporters in response to NaCl stress in one particular plant, the facultative halophyte Mesembryanthemum crystallinum Na(+)/H(+) antiporters of M. crystallinum were cloned by RACE-PCR from total mRNA of leaf mesophyll cells. Functional complementation of Saccharomyces cerevisiae and Escherichia coli mutants was performed. The kinetics of changes in the expression of antiporters were quantified by real-time PCR in leaves and roots. Five Na(+)/H(+) antiporters (McSOS1, McNhaD, McNHX1, McNHX2 and McNHX3) were cloned, representing the entire set of these transporters in M. crystallinum. The functionality of McSOS1, McHX1 and McNhaD was demonstrated in complementation experiments. Quantitative analysis revealed a temporal correlation between salt accumulation and expression levels of genes in leaves, but not in roots, which was most pronounced for McNhaD. Results suggest a physiological role of McSOS1, McNhaD and McNHX1 in Na(+) compartmentation during plant adaptation to high salinity. The study also provides evidence for salt-induced expression and function of the Na(+)/H(+) antiporter McNhaD in chloroplasts and demonstrates that the chloroplast is one of the compartments involved in the response of cells to salt stress.

  19. Emerged HA and NA mutants of the pandemic influenza H1N1 viruses with increasing epidemiological significance in Taipei and Kaohsiung, Taiwan, 2009-10.

    Directory of Open Access Journals (Sweden)

    Chuan-Liang Kao

    Full Text Available The 2009 influenza pandemic provided an opportunity to observe dynamic changes of the hemagglutinin (HA and neuraminidase (NA of pH1N1 strains that spread in two metropolitan areas--Taipei and Kaohsiung. We observed cumulative increases of amino acid substitutions of both HA and NA that were higher in the post-peak than in the pre-peak period of the epidemic. About 14.94% and 3.44% of 174 isolates had one and two amino acids changes, respective, in the four antigenic sites. One unique adaptive mutation of HA2 (E374K was first detected three weeks before the epidemic peak. This mutation evolved through the epidemic, and finally emerged as the major circulated strain, with significantly higher frequency in the post-peak period than in the pre-peak (64.65% vs 9.28%, p<0.0001. E374K persisted until ten months post-nationwide vaccination without further antigenic changes (e.g. prior to the highest selective pressure. In public health measures, the epidemic peaked at seven weeks after oseltamivir treatment was initiated. The emerging E374K mutants spread before the first peak of school class suspension, extended their survival in high-density population areas before vaccination, dominated in the second wave of class suspension, and were fixed as herd immunity developed. The tempo-spatial spreading of E374K mutants was more concentrated during the post-peak (p = 0.000004 in seven districts with higher spatial clusters (p<0.001. This is the first study examining viral changes during the naïve phase of a pandemic of influenza through integrated virological/serological/clinical surveillance, tempo-spatial analysis, and intervention policies. The vaccination increased the percentage of E374K mutants (22.86% vs 72.34%, p<0.001 and significantly elevated the frequency of mutations in Sa antigenic site (2.36% vs 23.40%, p<0.001. Future pre-vaccination public health efforts should monitor amino acids of HA and NA of pandemic influenza viruses isolated at

  20. Diffusion of propylene adsorbed in Na-Y and Na-ZSM5 zeolites ...

    Indian Academy of Sciences (India)

    Here we report the quasielastic neutron scattering and FTIR studies on the dynamics of propylene adsorbed in Na-Y and Na-ZSM5 zeolites. QENS data show that although the mechanism of translational motion of propylene is jump diffusion in both the cases of Na-Y and Na-ZSM5 zeolites, the diffusivity is affected by the ...

  1. Diffusion of propylene adsorbed in Na-Y and Na-ZSM5 zeolites ...

    Indian Academy of Sciences (India)

    Abstract. Here we report the quasielastic neutron scattering and FTIR studies on the dynamics of propylene adsorbed in Na-Y and Na-ZSM5 zeolites. QENS data show that although the mechanism of translational motion of propylene is jump diffusion in both the cases of Na-Y and Na-ZSM5 zeolites, the diffusivity is affected ...

  2. Na+,K+-ATPase Na+ affinity in rat skeletal muscle fiber types

    DEFF Research Database (Denmark)

    Kristensen, Michael; Juel, Carsten

    2010-01-01

    Previous studies in expression systems have found different ion activation of the Na(+)/K(+)-ATPase isozymes, which suggest that different muscles have different ion affinities. The rate of ATP hydrolysis was used to quantify Na(+),K(+)-ATPase activity, and the Na(+) affinity of Na(+),K(+)-ATPase...

  3. Dijagnostičko-terapijski pristup pacijentu sa sumnjom na alergiju na penicilin

    OpenAIRE

    Stanić Benić, Mirjana; Vlahović-Palčvevski, Vera

    2016-01-01

    Iako se ne zna točna učestalost alergije na lijekove, pripisuje joj se oko 1/10 svih neželjenih reakcija na lijekove. Neželjene reakcije na penicilin javljaju se u 0,7 – 10 % ljudi koji primaju penicilin. Stvarna učestalost alergije na penicilin još je manja, budući da su se u prošlosti mnoge nealergijske nuspojave antibiotika pripisivale alergiji. U pacijenata kod kojih postoji sumnja na alergiju na penicilin važno je utvrditi alergološki status. Nepotrebno izbjegavanje upotrebe penicilina, ...

  4. Alternative splice variant of the thiazide-sensitive NaCl cotransporter

    DEFF Research Database (Denmark)

    Tutakhel, Omar A Z; Jeleń, Sabina; Valdez-Flores, Marco

    2016-01-01

    The thiazide-sensitive NaCl cotransporter (NCC) is an important pharmacological target in the treatment of hypertension. Human SLC12A3 gene, encoding NCC, gives rise to three isoforms. Only the 3(rd) isoform has been extensively investigated. The aim of the present study was, therefore, to establ...

  5. Crowdsourcing: impactos na performance na venda de produtos

    OpenAIRE

    Alves, Milton Ruiz Rodrigues

    2015-01-01

    Vivemos num momento em que as expectativas de pensadores como Lévy (1998), se concretizaram. O consumidor mudou, passou a exigir um diálogo entre a empresa e a sociedade. Esse novo cenário cobrou da tecnologia formas de aperfeiçoar e agilizar este processo de comunicação, plataformas baseadas na internet que propiciem e incentivam a troca de informações. Nesta esfera surgiu o crowdsourcing, com a disponibilização do capital intelectual das massas, que acabou por mudar a form...

  6. As atitudes do terapeuta na teoria centrada na pessoa

    OpenAIRE

    Carreteiro, Tereza Cristina Othenio Cordeiro

    1981-01-01

    Este trabalho procede a um exame arqueológico das atitudes terapêuticas ao longo do desenvolvimento da teoria Rogeriana. Estuda-se o processo evolutivo que compõe o referido corpo teórico. Inicialmente, é feita uma breve apresentação de Carl Rogers, precursor da Abordagem Centrada na Pessoa. São discorridos aspectos pessoais e profissionais do autor assim como suas principais contribuições no campo da Psicologia clínica. A teoria é decomposta em três fases principais: pré história da empat...

  7. Priklop perifernih naprav na Raspberry Pi

    OpenAIRE

    Fabčič, Simon

    2015-01-01

    Moje diplomsko delo obravnava povezovanje perifernih naprav na Raspberry Pi. Najprej je predstavljena oprema, ki sem jo potreboval pri pisanju. Kot glavno strojno opremo sem uporabil enoploščni računalnik Raspberry Pi (model B) in nanj namestil operacijski sistem Raspbian. Na njem je bilo že pred naloženo razvojno okolje za programski jezik Python, ki sem ga tudi uporabljal. Nato sem prikazal, kako na Raspberry Pi na različne načine in prek raznih protokolov povežemo periferne naprave ter dig...

  8. Upravnopravna regulacija prava na pristup informacijama

    OpenAIRE

    Basta, Đurđica

    2016-01-01

    Pravo na pristup informacijama čini jedan od ključnih elemenata informacijskoga upravnog prava. U većini zemalja, uključujući Hrvatsku, pravo na pristup informacijama javnog sektora relativno je novo pravo. Uz Ustav RH kao temeljni dokument, od ožujka 2013. na snazi je novi, drugi hrvatski Zakon o pravu na pristup informacijama. Kritike na račun pravnog okvira kao i institucionalne podrške njegovoj provedbi doveli su do izmjena Zakona 2010. odnosno 2011., a 2013. i do donošenja...

  9. Physiological and molecular mechanisms mediating xylem Na(+) loading in barley in the context of salinity stress tolerance.

    Science.gov (United States)

    Zhu, Min; Zhou, Meixue; Shabala, Lana; Shabala, Sergey

    2017-07-01

    Time-dependent kinetics of xylem Na(+) loading was investigated using a large number of barley genotypes contrasting in their salinity tolerance. Salt-sensitive varieties were less efficient in controlling xylem Na(+) loading and showed a gradual increase in the xylem Na(+) content over the time. To understand underlying ionic and molecular mechanisms, net fluxes of Ca(2+) , K(+) and Na(+) were measured from the xylem parenchyma tissue in response to H2 O2 and ABA; both of them associated with salinity stress signalling. Our results indicate that NADPH oxidase-mediated apoplastic H2 O2 production acts upstream of the xylem Na(+) loading and is causally related to ROS-inducible Ca(2+) uptake systems in the root stelar tissue. It was also found that ABA regulates (directly or indirectly) the process of Na(+) retrieval from the xylem and the significant reduction of Na(+) and K(+) fluxes induced by bumetanide are indicative of a major role of chloride cation co-transporter (CCC) on xylem ion loading. Transcript levels of HvHKT1;5_like and HvSOS1_like genes in the root stele were observed to decrease after salt stress, while there was an increase in HvSKOR_like gene, indicating that these ion transporters are involved in primary Na(+) /K(+) movement into/out of xylem. © 2016 John Wiley & Sons Ltd.

  10. Variability in Measures of Exhaled Breath Na, Influence of pulmonary Blood Flow and salivary Na

    Directory of Open Access Journals (Sweden)

    Courtney M. Wheatley

    2010-04-01

    Full Text Available The assessment of inflammatory markers and ions in exhaled breath condensate (EBC is being utilized more frequently in diseases such as asthma and cystic fibrosis with marked variability in EBC measures, including those of exhaled Na + . We sought to determine if variability in exhaled Na + was due to differences in pulmonary blood flow (PBF or Na + in the mouth (salivary Na + . We measured exhaled Na + three times with coinciding sampling of salivary Na + and assessment of PBF (using acetylene rebreathing in 13 healthy subjects (54% female, age = 27 ± 7 yrs., ht. = 172 ± 10 cm, wt. = 70 ± 21 kg, BMI = 22 ± 7 kg/m 2 mean ± SD. Exhaled Na + averaged 2.7 ± 1.2 mmol/l, and salivary Na + averaged 5.51 ± 4.58 mmol/l. The coefficients of variation across all three measures in all 13 subjects averaged 30% for exhaled Na + and 83% for salivary Na + , within subjects the variability across the three measures averaged 30% for exhaled Na + and 38% for salivary Na + . Across all three measures in all 13 subjects the relationship between PBF and exhaled Na + averaged 0.027 ( P = 0.87, and the relationship between salivary Na + and exhaled Na + concentrations averaged 0.59 ( P = 0.001. Also, we sought to determine the relationship between exhaled Na + and serum Na + in an addition 20 subjects. There was a moderate and significant relationship between serum Na + and exhaled Na + (r = 0.37, P = 0.04. These findings suggest there that the variability in exhaled Na + is caused, at least in part, by droplet formation from within the mouth as turbulent air passes through and that there is a flux of ions from the pulmonary blood into the airways.

  11. Istraživanje uticaja nekih parametara na osetljivost eksplozivnih materija na toplotni impuls

    OpenAIRE

    Jeremić Radun

    2002-01-01

    Ispitivana je osetljivost nekoliko vrsta eksplozivnih materija na toplotni impuls merenjem vremena zadrške u zavisnosti od temperature. Na osnovu eksperimentalnih rezultata određene su vrednosti energije aktivacije i predeksponencijalni faktori u jednačini Semjonova. Istraživan je i uticaj mase ispitivanog uzorka, granulacije i veličine kristala eksploziva na osetljivost na toplotni impuls. Pokazano je da ovi parametri imaju značajan uticaj na osetljivost eksplozivnih materija, pa je neophodn...

  12. A new computational strategy for predicting essential genes.

    Science.gov (United States)

    Cheng, Jian; Wu, Wenwu; Zhang, Yinwen; Li, Xiangchen; Jiang, Xiaoqian; Wei, Gehong; Tao, Shiheng

    2013-12-21

    Determination of the minimum gene set for cellular life is one of the central goals in biology. Genome-wide essential gene identification has progressed rapidly in certain bacterial species; however, it remains difficult to achieve in most eukaryotic species. Several computational models have recently been developed to integrate gene features and used as alternatives to transfer gene essentiality annotations between organisms. We first collected features that were widely used by previous predictive models and assessed the relationships between gene features and gene essentiality using a stepwise regression model. We found two issues that could significantly reduce model accuracy: (i) the effect of multicollinearity among gene features and (ii) the diverse and even contrasting correlations between gene features and gene essentiality existing within and among different species. To address these issues, we developed a novel model called feature-based weighted Naïve Bayes model (FWM), which is based on Naïve Bayes classifiers, logistic regression, and genetic algorithm. The proposed model assesses features and filters out the effects of multicollinearity and diversity. The performance of FWM was compared with other popular models, such as support vector machine, Naïve Bayes model, and logistic regression model, by applying FWM to reciprocally predict essential genes among and within 21 species. Our results showed that FWM significantly improves the accuracy and robustness of essential gene prediction. FWM can remarkably improve the accuracy of essential gene prediction and may be used as an alternative method for other classification work. This method can contribute substantially to the knowledge of the minimum gene sets required for living organisms and the discovery of new drug targets.

  13. Silicene for Na-ion battery applications

    KAUST Repository

    Zhu, Jiajie

    2016-08-19

    Na-ion batteries are promising candidates to replace Li-ion batteries in large scale applications because of the advantages in natural abundance and cost of Na. Silicene has potential as the anode in Li-ion batteries but so far has not received attention with respect to Na-ion batteries. In this context, freestanding silicene, a graphene-silicene-graphene heterostructure, and a graphene-silicene superlattice are investigated for possible application in Na-ion batteries, using first-principles calculations. The calculated Na capacities of 954mAh/g for freestanding silicene and 730mAh/g for the graphenesilicene superlattice (10% biaxial tensile strain) are highly competitive and potentials of >0.3 V against the Na/Na potential exceed the corresponding value of graphite. In addition, the diffusion barriers are predicted to be <0.3 eV.

  14. Nax loci affect SOS1-like Na+/H+ exchanger expression and activity in wheat.

    Science.gov (United States)

    Zhu, Min; Shabala, Lana; Cuin, Tracey A; Huang, Xin; Zhou, Meixue; Munns, Rana; Shabala, Sergey

    2016-02-01

    Salinity stress tolerance in durum wheat is strongly associated with a plant's ability to control Na(+) delivery to the shoot. Two loci, termed Nax1 and Nax2, were recently identified as being critical for this process and the sodium transporters HKT1;4 and HKT1;5 were identified as the respective candidate genes. These transporters retrieve Na(+) from the xylem, thus limiting the rates of Na(+) transport from the root to the shoot. In this work, we show that the Nax loci also affect activity and expression levels of the SOS1-like Na(+)/H(+) exchanger in both root cortical and stelar tissues. Net Na(+) efflux measured in isolated steles from salt-treated plants, using the non-invasive ion flux measuring MIFE technique, decreased in the sequence: Tamaroi (parental line)>Nax1=Nax2>Nax1:Nax2 lines. This efflux was sensitive to amiloride (a known inhibitor of the Na(+)/H(+) exchanger) and was mirrored by net H(+) flux changes. TdSOS1 relative transcript levels were 6-10-fold lower in Nax lines compared with Tamaroi. Thus, it appears that Nax loci confer two highly complementary mechanisms, both of which contribute towards reducing the xylem Na(+) content. One enhances the retrieval of Na(+) back into the root stele via HKT1;4 or HKT1;5, whilst the other reduces the rate of Na(+) loading into the xylem via SOS1. It is suggested that such duality plays an important adaptive role with greater versatility for responding to a changing environment and controlling Na(+) delivery to the shoot. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  15. Regulation of Na+/K+ ATPase transport velocity by RNA editing.

    Directory of Open Access Journals (Sweden)

    Claudia Colina

    2010-11-01

    Full Text Available Because firing properties and metabolic rates vary widely, neurons require different transport rates from their Na(+/K(+ pumps in order to maintain ion homeostasis. In this study we show that Na(+/K(+ pump activity is tightly regulated by a novel process, RNA editing. Three codons within the squid Na(+/K(+ ATPase gene can be recoded at the RNA level, and the efficiency of conversion for each varies dramatically, and independently, between tissues. At one site, a highly conserved isoleucine in the seventh transmembrane span can be converted to a valine, a change that shifts the pump's intrinsic voltage dependence. Mechanistically, the removal of a single methyl group specifically targets the process of Na(+ release to the extracellular solution, causing a higher turnover rate at the resting membrane potential.

  16. Molecular characterization of glycoprotein genes and phylogenetic analysis of two swine paramyxoviruses isolated from United States.

    Science.gov (United States)

    Qiao, Dan; Janke, Bruce H; Elankumaran, Subbiah

    2009-08-01

    Two swine paramyxoviruses (SPMV)-(81-19252 (Texas-81) and 92-7783 (ISU-92)-were isolated from encephalitic pigs in the United States in 1981 and 1992. Antigenic, morphologic, and biological characteristics of these two viruses were essentially similar to members of the family Paramyxoviridae. Antigenic analysis by indirect fluorescent antibody, immunoblot, and one-way cross-neutralization tests placed these viruses along with bovine parainfluenza 3 (BPIV3) viruses. Purified virions were 50-300 nm in size and morphologically indistinguishable from other paramyxoviruses. These two viruses hemagglutinated red blood cells and had neuraminidase activity. The gene junctions of fusion (F) and hemagglutinin (HN) glycoprotein genes of these viruses contained highly conserved transcription start and stop signal sequences and trinucleotide intergenic regions similar to other Paramyxoviridae. The F gene of ISU-92 was longer than Texas-81 due to insertion of a 24-nucleotide "U"-rich 3' untranslated region. Structure-based sequence alignment of glycoproteins of these two SPMVs indicated that they are essentially similar in structure and function to parainfluenzaviruses. The Texas-81 strain was closely related to BPIV3 Shipping Fever (SF) strain at nucleotide and amino acid level, while the ISU-92 strain was more closely related to BPIV3 910N strain. The envelope glycoproteins of ISU-92 had only approximately 92 and approximately 96% identity at nucleotide and amino acid levels with BPIV3-SF strain, respectively. The high sequence identities to BPIV3 indicated cross-species infection in pigs. Phylogenetic analyses based on both F protein and HN protein suggested the classification of these viruses into the subfamily Paramyxovirinae, genus Respirovirus, and genotype A of BPIV3.

  17. Genetic variation in the HN and SH genes of mumps viruses: a comparison of strains from mumps cases with and without neurological symptoms.

    Science.gov (United States)

    Cui, Aili; Brown, David W G; Xu, Wenbo; Jin, Li

    2013-01-01

    It is known that mumps virus (MuV) strains may vary in their neurovirulent capacity, and certain MuV strains may be highly neurotropic. In animal models and epidemiological studies, mutations at specific amino acids (aa) have been proposed to be associated with neurovirulence. To assess whether these genetic variations can be observed in clinical samples from patients and if they correlate with neurovirulence as determined by clinical symptoms, 39 mumps patients with or without neurological symptoms were investigated. Respiratory samples, oral fluids, throat swabs, and neurological and cerebrospinal fluid samples were tested by RT-PCR and products sequenced. Sequences of the entire small hydrophobic (SH) gene and the partial hemagglutinin-neuraminidase (HN) gene were compared. The results showed there was no significant difference between the samples of the two groups of patients at the aa sites in either the HN protein or the SH protein, which have previously been hypothesized to be associated with neurovirulence or antigenicity. The occurrence of neurological symptoms of mumps does not appear to be due to a single point mutation in either the HN or SH gene.

  18. Genetic variation in the HN and SH genes of mumps viruses: a comparison of strains from mumps cases with and without neurological symptoms.

    Directory of Open Access Journals (Sweden)

    Aili Cui

    Full Text Available BACKGROUND: It is known that mumps virus (MuV strains may vary in their neurovirulent capacity, and certain MuV strains may be highly neurotropic. In animal models and epidemiological studies, mutations at specific amino acids (aa have been proposed to be associated with neurovirulence. To assess whether these genetic variations can be observed in clinical samples from patients and if they correlate with neurovirulence as determined by clinical symptoms, 39 mumps patients with or without neurological symptoms were investigated. PRINCIPAL FINDINGS: Respiratory samples, oral fluids, throat swabs, and neurological and cerebrospinal fluid samples were tested by RT-PCR and products sequenced. Sequences of the entire small hydrophobic (SH gene and the partial hemagglutinin-neuraminidase (HN gene were compared. CONCLUSIONS: The results showed there was no significant difference between the samples of the two groups of patients at the aa sites in either the HN protein or the SH protein, which have previously been hypothesized to be associated with neurovirulence or antigenicity. The occurrence of neurological symptoms of mumps does not appear to be due to a single point mutation in either the HN or SH gene.

  19. Phylogenetic analysis of influenza A viruses (H3N2 circulating in Zhytomyr region during 2013–2014 epidemic season

    Directory of Open Access Journals (Sweden)

    Boyalska O. G.

    2015-06-01

    Full Text Available Aim. To perform phylogenetic analysis of the hemagglutinin (HA and neuraminidase (NA genes of influenza A(H3N2 viruses circulating in the Zhytomyr region during 2013–2014 epidemic season. To make comparison of the HA and NA genes sequences of the Zhytomyr region isolates with the HA and NA genes sequences of influenza viruses circulating in the world. Methods. Laboratory diagnosis was conducted by real-time polymerase chain reaction (RT-PCR. In this study the sequencing and phylogenetic analysis were carried out. Results. For the first time the genes of influenza A(H3N2 viruses isolated in the Zhytomyr region during 2013–2014 epidemic season, coding hemagglutinin and neuraminidase were compared with their orthologs. According to the results of this comparison the phylogenetic tree was constructed. Additionally, the amino acid substitutions of the influenza viruses circulating in Ukraine and worldwide were analyzed. Conclusions. The nucleotide sequences of the influenza A(H3N2 viruses genes HA and NA isolated in the Zhytomyr region were identified. Based on the nucleotide sequences of HA and NA we constructed the influenza virus phylogenetic tree demonstrating that the virus isolated in the Zhytomyr region was closely related to the Ukrainian isolate from Kharkov and in the world to the isolates from Germany, Romania, Italy.

  20. Gene Ontology

    Directory of Open Access Journals (Sweden)

    Gaston K. Mazandu

    2012-01-01

    Full Text Available The wide coverage and biological relevance of the Gene Ontology (GO, confirmed through its successful use in protein function prediction, have led to the growth in its popularity. In order to exploit the extent of biological knowledge that GO offers in describing genes or groups of genes, there is a need for an efficient, scalable similarity measure for GO terms and GO-annotated proteins. While several GO similarity measures exist, none adequately addresses all issues surrounding the design and usage of the ontology. We introduce a new metric for measuring the distance between two GO terms using the intrinsic topology of the GO-DAG, thus enabling the measurement of functional similarities between proteins based on their GO annotations. We assess the performance of this metric using a ROC analysis on human protein-protein interaction datasets and correlation coefficient analysis on the selected set of protein pairs from the CESSM online tool. This metric achieves good performance compared to the existing annotation-based GO measures. We used this new metric to assess functional similarity between orthologues, and show that it is effective at determining whether orthologues are annotated with similar functions and identifying cases where annotation is inconsistent between orthologues.

  1. Tetrodotoxin-resistant voltage-gated sodium channels Na(v)1.8 and Na(v)1.9 are expressed in the retina.

    Science.gov (United States)

    O'Brien, Brendan J; Caldwell, John H; Ehring, George R; Bumsted O'Brien, Keely M; Luo, Songjiang; Levinson, S Rock

    2008-06-20

    Voltage-gated sodium channels (VGSCs) are one of the fundamental building blocks of electrically excitable cells in the nervous system. These channels are responsible for the generation of action potentials that are required for the communication of neuronal signals over long distances within a cell. VGSCs are encoded by a family of nine genes whose products have widely varying biophysical properties. In this study, we have detected the expression of two atypical VGSCs (Na(v)1.8 and Na(v)1.9) in the retina. Compared with more common VGSCs, Na(v)1.8 and Na(v)1.9 have unusual biophysical and pharmacological properties, including persistent sodium currents and resistance to the canonical sodium channel blocker tetrodotoxin (TTX). Our molecular biological and immunohistochemical data derived from mouse (Mus musculus) retina demonstrate expression of Na(v)1.8 by retinal amacrine and ganglion cells, whereas Na(v)1.9 is expressed by photoreceptors and Müller glia. The fact that these channels exist in the central nervous system (CNS) and exhibit robust TTX resistance requires a re-evaluation of prior physiological, pharmacological, and developmental data in the visual system, in which the diversity of VGSCs has been previously underestimated. (c) 2008 Wiley-Liss, Inc.

  2. Na+,K+-pump stimulation improves contractility in isolated muscles of mice with hyperkalemic periodic paralysis

    Science.gov (United States)

    Nielsen, Ole Bækgaard; Clausen, Johannes D.; Pedersen, Thomas Holm; Hayward, Lawrence J.

    2011-01-01

    In patients with hyperkalemic periodic paralysis (HyperKPP), attacks of muscle weakness or paralysis are triggered by K+ ingestion or rest after exercise. Force can be restored by muscle work or treatment with β2-adrenoceptor agonists. A missense substitution corresponding to a mutation in the skeletal muscle voltage-gated Na+ channel (Nav1.4, Met1592Val) causing human HyperKPP was targeted into the mouse SCN4A gene (mutants). In soleus muscles prepared from these mutant mice, twitch, tetanic force, and endurance were markedly reduced compared with soleus from wild type (WT), reflecting impaired excitability. In mutant soleus, contractility was considerably more sensitive than WT soleus to inhibition by elevated [K+]o. In resting mutant soleus, tetrodotoxin (TTX)-suppressible 22Na uptake and [Na+]i were increased by 470 and 58%, respectively, and membrane potential was depolarized (by 16 mV, P muscles of mutant mice, the content of Na+,K+ pumps was 28, 62, and 33% higher than in WT, respectively, possibly reflecting the stimulating effect of elevated [Na+]i on the synthesis of Na+,K+ pumps. The results confirm that the functional disorders of skeletal muscles in HyperKPP are secondary to increased Na+ influx and show that contractility can be restored by acute stimulation of the Na+,K+ pumps. Calcitonin gene-related peptide (CGRP) restored force in mutant soleus but caused no detectable increase in 86Rb uptake. Repeated excitation and capsaicin also restored contractility, possibly because of the release of endogenous CGRP from nerve endings in the isolated muscles. These observations may explain how mild exercise helps locally to prevent severe weakness during an attack of HyperKPP. PMID:21708955

  3. VPLIV VODENJA NA RAZVOJ ORGANIZACIJE

    OpenAIRE

    Rauš, Ivo

    2010-01-01

    Ključno vlogo pri razvoju človeških potencialov in sproščanju posameznikovih notranjih zmožnosti imajo njihovi neposredni vodje. Le ti naj bi skrbeli za ustvarjanje takega delovnega okolja, ki bo spodbujalo ustvarjalne dosežke, inovativnost, izvirnost idej, pripadnost in timsko sodelovanje. Vodenje zavzema prvo mesto na lestvici ključnih managerskih izzivov sedanjosti in srednjeročne prihodnosti, o čemer govorijo tudi številne zadnje raziskave ameriških in evropskih strokovnjakov za razvoj...

  4. Chiaroscuro: a luz na sombra

    OpenAIRE

    Flamínio Jallageas de Lima Cardoso

    2015-01-01

    Este ensaio objetiva refletir sobre minha série fotográfica Chiaroscuro, na qual trabalho constrastes entre luz e sombra e as consequentes representações de vazio e memória. Para tanto, elejo os teóricos da antropologia da imagem Jacques Rancière, Georges Didi-Huberman, Victor Stoichita e Hans Belting como meus interlocutores. Uma vez que um dos propósitos deste trabalho abrange observar possíveis inter-relações entre a série e alguns artistas contemporâneos, estabeleço reflexões acerca da pr...

  5. Na alvorada de um sport

    OpenAIRE

    Sartori, Carina

    2013-01-01

    Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Filosofia e Ciências Humanas, Programa de Pós-graduação em História, Florianópolis, 2013 Em Florianópolis, a primeira Sociedade de Regatas organizada, meados do século XIX, contaria com a participação dos Coronéis da Marinha e da Escola de Menores e Marinheiros da Canhoneira. Na virada do século, a fundação do Clube 29 de Abril permitiria que as atividades relacionadas ao Remo fossem oferecidas a todos que se associ...

  6. A virtude na psicologia positiva

    OpenAIRE

    Cardoso, José Augusto Rento

    2014-01-01

    O presente trabalho de psicologia teórica trata-se de um estudo crítico do conceito de virtude na Psicologia Positiva. Definida como movimento por alguns e como um novo campo por outros, a Psicologia Positiva reúne sob um mesmo “guarda-chuva” diversos estudos, passados e presentes, de várias áreas da Psicologia, como a Psicologia da Personalidade, a Psicologia Social, a Psicologia do Desenvolvimento e a Psicologia Moral. No presente trabalho, a mesma é compreendida como uma perspectiva e um m...

  7. Tortura : na atividade policial investigativa

    OpenAIRE

    Giudice de Argollo, Helvécio

    2003-01-01

    Este trabalho versa sobre o fenômeno social da tortura; do seu sentido polissêmico, do seu aspecto multifacetado e dos desafios que engendra, notadamente da sua perseverança nas sociedades, a despeito do banimento pelos textos legais, tanto no plano do direito transnacional quanto na maioria dos ordenamentos jurídicos do mundo ocidental. A tortura tem uma história que se confunde com a história da própria humanidade e, nesse sentido, o seu estudo desafia um périplo histórico...

  8. The NA62 RICH detector

    Energy Technology Data Exchange (ETDEWEB)

    Lenti, Massimo, E-mail: lenti@fi.infn.i [Sezione dell' INFN di Firenze, I-50019 Sesto Fiorentino (Italy)

    2011-05-21

    The NA62 RICH must separate charged pions from muons in a 15-35 GeV/c momentum range. The detector will have a 17 m long, 4 m wide cylindrical vessel, filled with neon at an atmospheric pressure. A mosaic of 20 mirrors, with 17 m focal length will be placed at the downstream end of the vessel while 2000 single anode photomultipliers will be placed at the upstream part. Two test beams were held at CERN in 2007 and 2009 with a RICH prototype. A muon suppression factor greater than 100 was validated.

  9. New results from NA49

    CERN Document Server

    Afanasiev, S V; Bächler, J; Barna, D; Barnby, L S; Bartke, Jerzy; Barton, R A; Behler, M; Betev, L; Bialkowska, H; Billmeier, A; Blume, C; Blyth, C O; Boimska, B; Botje, M; Bracinik, J; Brady, F P; Bramm, R; Brun, R; Buncic, P; Carr, L; Cebra, D; Cerny, V; Cooper, G E; Cramer, J G; Csató, P; Dinkelaker, P; Eckardt, V; Eckhardt, F; Ferenc, D; Filip, P; Fischer, H G; Foder, Z; Foka, P Y; Freund, P; Friese, V; Gál, J; Ganz, R E; Gazdzicki, M; Georgopoulos, G; Gladysz-Dziadus, E; Harris, J W; Hegyi, S; Höhne, C; Igo, G; Jacobs, P; Jones, P G; Kadija, K; Kolesnikov, V I; Kollegger, T; Kowalski, M; Kraus, I; Kreps, M; Lasiuk, B; Van Leeuwen, M; Lévai, Peter; Malakhov, A I; Margetis, S; Markert, C; Mayes, B W; Melkumov, G L; Mischke, A; Molnár, J; Nelson, J M; Odyniec, Grazyna Janina; Oldenburg, M; Pálla, G; Panagiotou, A D; Perl, K; Petridis, A; Pikna, M; Pinsky, L; Poskanzer, A M; Prindle, D J; Pühlhofer, F; Putschke, J; Reid, J G; Renfordt, A; Retyk, W; Ritter, H G; Röhrich, D; Roland, C; Roland, G; Rybicki, A; Sammer, T; Sann, H; Schäfer, E; Schmitz, N; Seyboth, P; Siklér, F; Sitár, B; Skrzypczak, E; Snellings, R; Squier, G T A; Stock, Reinhard; Ströbele, H; Susa, T; Szentpétery, I; Sziklai, J; Toy, M; Trainor, T A; Trentalange, S; Ullrich, T S; Varga, D; Vassiliou, Maria; Veres, G I; Vesztergombi, G; Voloshin, S A; Vranic, D; Wang, F; Weerasundara, D D; Wenig, S; Wetzler, A; Whitten, C; Xu, N; Yates, T A; Koo, I K; Zaranek, J; Zimányi, J

    2002-01-01

    Recent results of the NA49 experiment are presented. These cover first results on pion and kaon production, HBT, and charge fluctuations from Pb+Pb reactions at 40 AGeV and their comparison to 158 AGeV beam energy. Furthermore a study on baryon number transfer in p+p, centrality selected p+Pb and Pb+Pb collisions at 158 AGeV and new results on the system size dependence of kaon yields, including C+C and Si+Si data, are presented. Additionally, a first result on Lambda Lambda correlations is shown. (11 refs).

  10. Perfil de sensibilidade antimicrobiana e detecção do gene ISS pela reação em cadeia da polimerase na tipificação de Escherichia coli patogênica em codornas de corte sob inspeção sanitária Profile of antimicrobial resistance and detection of iss gene by the polymerase chain reaction in the typification of pathogenic Escherichia coli in meat type quails under sanitary inspection

    Directory of Open Access Journals (Sweden)

    Dayse Lima da Costa Abreu

    2010-05-01

    Full Text Available A patogenicidade das cepas de Escherichia coli está relacionada à expressão de fatores de virulência encontrados em elementos genéticos denominados plasmídios. O patotipo APEC, responsável por diferentes tipos de doenças em aves, pode apresentar o gene iss que aumenta a resistência das cepas de E. coli aos efeitos líticos do soro, além da resistência a diversos antimicrobianos. Este estudo foi conduzido para detectar E. coli em traquéias de codornas destinadas ao abate e avaliar, pela presença do gene iss e o perfil de susceptibilidade antimicrobiana, o potencial patogênico para aves e humanos dos isolados obtidos. Foram coletadas 180 traquéias de codornas para detecção de E. coli, determinação do perfil de resistência a agentes antimicrobianos e posterior detecção, por reação em cadeia da polimerase (PCR, do gene iss. Das traquéias analisadas, 8,9 % (16/180 foram positivas para E. coli, sendo obtidos 20 isolados deste agente. A maioria dos isolados foi resistente à Tetraciclina (16/20, seguida pela Ceftazidima (13/20 e Ácido Nalidíxico (12/20, sendo apenas um resistente à Amoxicilina. A detecção do gene iss ocorreu em 55% (11/20 dos isolados. A presença do gene iss e a resistência a múltiplos antimicrobianos dos isolados obtidos neste estudo pode indicar um possível potencial patogênico das cepas de E. coli tanto para codornas quanto para outros tipos de aves e animais e mesmo para o ser humano que fique em contato com as mesmas.The pathogenicity of Escherichia coli strains is partially related to the expression of virulence factors genes, present in genetic elements called plasmids. APEC strains responsible for diseases in birds may present the iss gene which increases the resistance of E. coli strains to the lityc effect of the host's serum, besides resistance to several antimicrobials. This study was conduced in order to detect E. coli in tracheae of meat-type quails and to evaluate, by the presence of

  11. Capacidade combinatória e ação gênica na expressão de caracteres de importância econômica em pimentão Combining ability and gene action in the expression of economically important traits in sweet pepper

    Directory of Open Access Journals (Sweden)

    Ildon Rodrigues do Nascimento

    2004-04-01

    Full Text Available Objetivou-se com o presente trabalho avaliar a capacidade combinatória de linhagens-elite de pimentão (Capsicum annuum L. e estudar os tipos de ação gênica envolvidos no controle de alguns dos caracteres de importância econômica, na tentativa de se identificar linhagens com potencial de originar híbridos competitivos com os atualmente plantados em escala comercial. O ensaio foi constituído de 30 híbridos, dos quais: (a 18 obtidos num esquema dialélico Norte Carolina II, a partir do cruzamento de dois grupos de genitores [grupo I - genitores femininos: L-005, PIM-013, PIM-014, Magda, PIX-021G_0818pl#01 e PIX-022E31pl#14; grupo II - genitores masculinos L-004, L-006 e MYR-29]; (b cinco híbridos adicionais F1(Hércules x L-004, F1(Itapetininga x L-004, F1(L-3509 Frutos amarelos x L-004, F1(PIX-021F0818 x L-006 e F1(PIX-023E39 x L-006; (c três híbridos entre os testadores F1(L-006 x L-004, F1(MYR-29 x L-004 e F1(MYR-29 x L-006; (d quatro híbridos comerciais (Magali R, Magali, Fortuna Super e Atenas. Utilizou-se o delineamento em blocos casualizados com 4 repetições. Avaliaram-se os seguintes caracteres: produção total, peso médio de frutos da produção total, produção precoce, comprimento de frutos, estrias nos frutos, profundidade de inserção do pedúnculo e altura de plantas. Houve predomínio dos efeitos gênicos aditivos para todas as características avaliadas, com exceção da produção total de frutos, em que os efeitos gênicos não-aditivos foram também importantes. Considerando os efeitos gênicos aditivos e não-aditivos conjuntamente, as melhores combinações foram F1(PIM-013 x MYR-29 e F1(PIX-022E31pl#14 x MYR-29, tanto para produção quanto para caracteres do fruto.This paper reports on the combining ability and gene action among elite inbred lines of sweet pepper (Capsicum annuum L., in an attempt to identify inbreds with good potencial to generate competitive commercial hybrids. The trial included 30

  12. Screening Reliable Reference Genes for RT-qPCR Analysis of Gene Expression in Moringa oleifera.

    Directory of Open Access Journals (Sweden)

    Li-Ting Deng

    Full Text Available Moringa oleifera is a promising plant species for oil and forage, but its genetic improvement is limited. Our current breeding program in this species focuses on exploiting the functional genes associated with important agronomical traits. Here, we screened reliable reference genes for accurately quantifying the expression of target genes using the technique of real-time quantitative polymerase chain reaction (RT-qPCR in M. oleifera. Eighteen candidate reference genes were selected from a transcriptome database, and their expression stabilities were examined in 90 samples collected from the pods in different developmental stages, various tissues, and the roots and leaves under different conditions (low or high temperature, sodium chloride (NaCl- or polyethyleneglycol (PEG- simulated water stress. Analyses with geNorm, NormFinder and BestKeeper algorithms revealed that the reliable reference genes differed across sample designs and that ribosomal protein L1 (RPL1 and acyl carrier protein 2 (ACP2 were the most suitable reference genes in all tested samples. The experiment results demonstrated the significance of using the properly validated reference genes and suggested the use of more than one reference gene to achieve reliable expression profiles. In addition, we applied three isotypes of the superoxide dismutase (SOD gene that are associated with plant adaptation to abiotic stress to confirm the efficacy of the validated reference genes under NaCl and PEG water stresses. Our results provide a valuable reference for future studies on identifying important functional genes from their transcriptional expressions via RT-qPCR technique in M. oleifera.

  13. Screening Reliable Reference Genes for RT-qPCR Analysis of Gene Expression in Moringa oleifera.

    Science.gov (United States)

    Deng, Li-Ting; Wu, Yu-Ling; Li, Jun-Cheng; OuYang, Kun-Xi; Ding, Mei-Mei; Zhang, Jun-Jie; Li, Shu-Qi; Lin, Meng-Fei; Chen, Han-Bin; Hu, Xin-Sheng; Chen, Xiao-Yang

    2016-01-01

    Moringa oleifera is a promising plant species for oil and forage, but its genetic improvement is limited. Our current breeding program in this species focuses on exploiting the functional genes associated with important agronomical traits. Here, we screened reliable reference genes for accurately quantifying the expression of target genes using the technique of real-time quantitative polymerase chain reaction (RT-qPCR) in M. oleifera. Eighteen candidate reference genes were selected from a transcriptome database, and their expression stabilities were examined in 90 samples collected from the pods in different developmental stages, various tissues, and the roots and leaves under different conditions (low or high temperature, sodium chloride (NaCl)- or polyethyleneglycol (PEG)- simulated water stress). Analyses with geNorm, NormFinder and BestKeeper algorithms revealed that the reliable reference genes differed across sample designs and that ribosomal protein L1 (RPL1) and acyl carrier protein 2 (ACP2) were the most suitable reference genes in all tested samples. The experiment results demonstrated the significance of using the properly validated reference genes and suggested the use of more than one reference gene to achieve reliable expression profiles. In addition, we applied three isotypes of the superoxide dismutase (SOD) gene that are associated with plant adaptation to abiotic stress to confirm the efficacy of the validated reference genes under NaCl and PEG water stresses. Our results provide a valuable reference for future studies on identifying important functional genes from their transcriptional expressions via RT-qPCR technique in M. oleifera.

  14. Gene Expression in the Human Endolymphatic Sac

    DEFF Research Database (Denmark)

    Møller, Martin Nue; Kirkeby, Svend; Vikeså, Jonas

    2015-01-01

    OBJECTIVES/HYPOTHESIS: The purpose of the present study is to explore, demonstrate, and describe the expression of genes related to the solute carrier (SLC) molecules of ion transporters in the human endolymphatic sac. STUDY DESIGN: cDNA microarrays and immunohistochemistry were used for analyses......a1 sodium-bicarbonate transporter, SLC9a2 sodium-hydrogen transporter, SLC12a3 thiazide-sensitive Na-Cl transporter, and SLC34a2 sodium-phosphate transporter. CONCLUSIONS: Several important ion transporters of the SLC family are expressed in the human endolymphatic sac, including Pendrin......, the thiazide-sensitive Na-Cl transporter, and the Na-phosphate transporter SLC34a2. The data provide a new knowledge base considering the ion-dependent metabolic mechanisms maintaining inner ear homeostasis. More specifically, the results indicate a strong similarity with the ion transportation occurring...

  15. Natural variants of AtHKT1 enhance Na+ accumulation in two wild populations of Arabidopsis.

    Directory of Open Access Journals (Sweden)

    Ana Rus

    2006-12-01

    Full Text Available Plants are sessile and therefore have developed mechanisms to adapt to their environment, including the soil mineral nutrient composition. Ionomics is a developing functional genomic strategy designed to rapidly identify the genes and gene networks involved in regulating how plants acquire and accumulate these mineral nutrients from the soil. Here, we report on the coupling of high-throughput elemental profiling of shoot tissue from various Arabidopsis accessions with DNA microarray-based bulk segregant analysis and reverse genetics, for the rapid identification of genes from wild populations of Arabidopsis that are involved in regulating how plants acquire and accumulate Na(+ from the soil. Elemental profiling of shoot tissue from 12 different Arabidopsis accessions revealed that two coastal populations of Arabidopsis collected from Tossa del Mar, Spain, and Tsu, Japan (Ts-1 and Tsu-1, respectively, accumulate higher shoot levels of Na(+ than do Col-0 and other accessions. We identify AtHKT1, known to encode a Na(+ transporter, as being the causal locus driving elevated shoot Na(+ in both Ts-1 and Tsu-1. Furthermore, we establish that a deletion in a tandem repeat sequence approximately 5 kb upstream of AtHKT1 is responsible for the reduced root expression of AtHKT1 observed in these accessions. Reciprocal grafting experiments establish that this loss of AtHKT1 expression in roots is responsible for elevated shoot Na(+. Interestingly, and in contrast to the hkt1-1 null mutant, under NaCl stress conditions, this novel AtHKT1 allele not only does not confer NaCl sensitivity but also cosegregates with elevated NaCl tolerance. We also present all our elemental profiling data in a new open access ionomics database, the Purdue Ionomics Information Management System (PiiMS; http://www.purdue.edu/dp/ionomics. Using DNA microarray-based genotyping has allowed us to rapidly identify AtHKT1 as the casual locus driving the natural variation in shoot Na

  16. O ATLETISMO NA PERSPECTIVA EDUCACIONAL

    Directory of Open Access Journals (Sweden)

    Leandro Araujo de Sousa

    2017-06-01

    Full Text Available A educação física escolar deve proporcionar experiências aos alunos para o seu desenvolvimento integral. As aulas muitas vezes são reduzidas ao ensino de esportes tradicionais, o que limita o repertório de movimentos dos alunos. Dessa forma o presente estudo de revisão objetiva problematizar o atletismo no ambiente escolar e propor possibilidades pedagógicas para seu ensino. Nesta pesquisa propomos tratar o ensino do atletismo na escola, caracterizando-o pelo seu repertório de movimentos que são naturais ao ser humano, como correr, saltar e lançar, o que possibilita maior praticidade em sua aplicação pedagógica, porém essas possibilidades não são consideradas no ambiente educacional. Diante dos dados encontrados, sugerimos que esse esporte seja abordado de forma multidimensional no ambiente escolar contextualizando-o à visão de algumas correntes teóricas. Nessa perspectiva do esporte educacional, o atletismo pode, assim como deve ser trabalhado nas aulas de educação física como instrumento pedagógico de ensino e auxiliar na formação do aluno de forma global.

  17. Identidades Homossexuais na Territorialidade Tradicionalista

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    Edipo Djavan dos Reis Göergen

    2015-12-01

    Full Text Available Este artigo apresenta o debate teórico que sustenta a pesquisa de mestrado desenvolvida pelo autor. Tendo por base suas vivências em contextos de tradicionalismo gaúcho, o autor deste trabalho tem percebido a grande quantidade de homossexuais, ou indivíduos que se auto-denominam não-heterossexuais, integrando as atividades artísticas do Movimento Tradicionalista Gaúcho, ou a territorialidade tradicionalista. Interessado em investigar tal fenômeno, o autor tem desenvolvido o projeto de pesquisa “Os Espaços Paradoxais de Relações Homoeróticas na Territorialidade Tradicionalista”, sob orientação do prof. Dr. Benhur Pinós da Costa, no Programa de Pós-Graduação em Geografia (UFSM. Para que essa pesquisa seja realizada, levam-se em conta as relações de gênero e de sexualidade que estão envolvidas no tema. Dessa forma, por ainda se tratarem de temáticas tidas como insignificantes na sociedade em geral e marginalizadas no campo científico, para que o presente trabalho fosse empreendido, buscou-se amparo teórico nas geografias feministas e queer, segmentos contemporâneos da ciência geográfica.

  18. Micotoxinas e Micotoxicoses na Avicultura

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    Santurio JM

    2000-01-01

    Full Text Available Esta revisão tem como objetivo principal mostrar, baseado em dezenas de pesquisas realizadas, os efeitos tóxicos das micotoxinas aflatoxinas, tricotecenos, zealenona e fumonisinas sobre o desempenho das aves. O descobrimento das propriedades hepatotóxicas e hepatocarcinogênicas de algumas linhagens de Aspergillus flavus e A. parasiticus em perus, na Inglaterra, no início da década de 1960, seguida pela elucidação da estrutura de seus metabólitos tóxicos, as aflatoxinas, deu novo enfoque e prioridade para a pesquisa sobre micotoxinas. Análises de aflatoxinas realizadas no Laboratório de Análises Micotoxicológicas (LAMIC da Universidade Fedaral de Santa Maria, entre os anos de 1986 e janeiro de 2000, em 15.600 amostras de alimentos destinados principalmente ao consumo animal, demonstram que no milho analisado, 41,9% das amostras estavam contaminadas por aflatoxinas. Em surtos de aflatoxicose no campo, uma das características mais marcantes é a má absorção que se manifesta como partículas de ração mal digeridas na excreta das aves. Também observa-se, em frangos e poedeiras que recebem AFL, extrema palidez das mucosas e pernas. Dietas deficientes em riboflavina ou colecalciferol (vit. D tornaram frangos sensíveis, nos índices de desenvolvimento corporal, a concentrações muito baixas de AFL. O efeito aflatoxina nos frangos é maior na fase inicial de crescimento, ou seja, quando as aves ingeriram aflatoxina nos primeiros 21 dias de vida, e quanto maior o nível de stress do lote, menor a quantidade de AFL para afetar negativamente seu desempenho, seja na produção de carne ou de ovos. As principais micotoxinas do grupo dos tricotecenos são: toxina T-2; deoxynivalenol (DON; diacetoxyscirpenol (DAS, todas produzidas através de diversas espécies de fungos do gênero Fusarium. Além dos tricotecenos, o fusarium também pode produzir zearalenona e fumonisinas. Dessas fusarium-toxinas, somente toxina T-2 gera patologias s

  19. Growth of binary organic NLO crystals: m.NA-p.NA and m.NA-CNA system

    Science.gov (United States)

    Singh, N. B.; Henningsen, T.; Hopkins, R. H.; Mazelsky, R.

    1993-01-01

    Experiments were carried out to grow 3.Nitroaniline (m.NA) crystals doped with 4.Nitroaniline (p.NA) and 2.chloro 4.Nitroaniline (CNA). The measured undercooling for m.NA, p.NA, and CNA were 0.21 tm K, 0.23 tm K, and 0.35 tm K respectively, where tm represents the melting temperature of the pure component. Because of the crystals' large heat of fusion and large undercooling, it was not possible to grow good quality crystals with low thermal gradients. In the conventional two-zone Bridgman furnace we had to raise the temperature of the hot zone above the decomposition temperature of CNA, p.NA, and m.NA to achieve the desired thermal gradient. To avoid decomposition, we used an unconventional Bridgman furnace. Two immiscible liquids, silicone oil and ethylene glycol, were used to build a special two-zone Bridgman furnace. A temperature gradient of 18 K/cm was achieved without exceeding the decomposition temperature of the crystal. The binary crystals, m.NA-p.NA and m.NA-CNA, were grown in centimeter size in this furnace. X-ray and optical characterization showed good optical quality.

  20. Fotografia na Imprensa: conflitos na câmara escura

    Directory of Open Access Journals (Sweden)

    José Lúcio da Silva Menezes

    2014-07-01

    Full Text Available Este artigo recupera o teor ideológico do discurso que a Revista Veja elaborou sobre a classe trabalhadora que emergiu no cenário brasileiro, no final da década de 1970. O destaque dado aos trabalhadores como temática central resgatada, deve-se aos movimentos grevistas por eles encetados, após anos de ocultamento de sua resistência sob a repressão da ditadura civil-militar (1964-1985. Com a rebeldia operária, as matérias a respeito deste grupo, na Revista,foram se tornando mais constantes e visíveis, pois já não era mais possível, para a Revista Veja, ignorar a existência da classe operária que, com a força insurgente das greves, ocupava as ruas do Brasil.

  1. A retrotransposon in an HKT1 family sodium transporter causes variation of leaf Na+ exclusion and salt tolerance in maize.

    Science.gov (United States)

    Zhang, Ming; Cao, Yibo; Wang, Zhiping; Wang, Zhi-Qiang; Shi, Junpeng; Liang, Xiaoyan; Song, Weibin; Chen, Qijun; Lai, Jinsheng; Jiang, Caifu

    2017-11-15

    Soil salinity is one of several major abiotic stresses that constrain maize productivity worldwide. An improved understanding of salt-tolerance mechanisms will thus enhance the breeding of salt-tolerant maize and boost productivity. Previous studies have indicated that the maintenance of leaf Na+ concentration is essential for maize salt tolerance, and the difference in leaf Na+ exclusion has previously been associated with variation in salt tolerance between maize varieties. Here, we report the identification and functional characterization of a maize salt-tolerance quantitative trait locus (QTL), Zea mays Na+ Content1 (ZmNC1), which encodes an HKT-type transporter (designated as ZmHKT1). We show that a natural ZmHKT1 loss-of-function allele containing a retrotransposon insertion confers increased accumulation of Na+ in leaves, and salt hypersensitivity. We next show that ZmHKT1 encodes a plasma membrane-localized Na+ -selective transporter, and is preferentially expressed in root stele (including the parenchyma cells surrounding the xylem vessels). We also show that loss of ZmHKT1 function increases xylem sap Na+ concentration and causes increased root-to-shoot Na+ delivery, indicating that ZmHKT1 promotes leaf Na+ exclusion and salt tolerance by withdrawing Na+ from the xylem sap. We conclude that ZmHKT1 is a major salt-tolerance QTL and identifies an important new gene target in breeding for improved maize salt tolerance. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

  2. Relative contributions of Na+/H+ exchange and Na+/HCO3- cotransport to ischemic Na-i(+) overload in isolated rat hearts

    NARCIS (Netherlands)

    Ten Hove, M; Nederhoff, MGJ; Van Echteld, CJA

    The Na+/H+ exchanger (NHE) and/or the Na+/HCO3- cotransporter (NBC) were blocked during ischemia in isolated rat hearts. Intracellular Na+ concentration ([Na+](i)), intracellular pH (pH(i)), and energy-related phosphates were measured by using simultaneous Na-23 and P-31 NMR spectroscopy. Hearts

  3. O Tempo e o Cuidado na velhice

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    Wanda Pereira Patrocinio

    2016-01-01

    Full Text Available Os temas deste Volume Temático da Revista Kairós Gerontologia versam sobre o multifacetado tema do “Cuidar” na velhice, em diferentes contextos e dentro de uma abordagem multidisciplinar. Com isso, espera-se que os textos aqui apresentados possam contribuir para o campo da pesquisa e da prática na área da Gerontologia, auxiliando profissionais e familiares na complexa tarefa de cuidar de idosos.  

  4. NaLS–ascorbic acid sys

    Indian Academy of Sciences (India)

    Unknown

    Abstract. The photogalvanic effect has been studied in three systems using photogalvanic cells and. NaLS–ascorbic acid–azur A, NaLS–ascorbic acid–azur B, NaLS–ascorbic acid–azur C systems. The pho- topotential and photocurrent generated by these systems are 770⋅0, 971⋅0, 623⋅0 mV and 160⋅0, 185⋅0,. 145⋅0 ...

  5. Clonagem de uma seqüência completa de cDNA e gene de Coffea arabica codificando uma proteína homóloga ao fator de tradução SUI1 de levedura: análise da expressão em órgãos da planta

    OpenAIRE

    Gaborit, Charlotte; Caillet, Victoria; Deshayes, Alain; Marraccini, Pierre

    2003-01-01

    A full-length cDNA (CaSUI1) was isolated from a Coffea arabica cDNA library from beans during maturation. Its putative translational product is highly homologous to the SUI1 protein of Saccharomyces cerevisiae which functions in concert with eIF2 and the initiator tRNA-Methionin in directing the ribosome to the proper start site of translation. The corresponding gene from coffee was also cloned and sequenced. Its organization is very similar to what observed for the same gene from rice (Oryza...

  6. The Role of Na+ and K+ Transporters in Salt Stress Adaptation in Glycophytes

    Directory of Open Access Journals (Sweden)

    Dekoum V. M. Assaha

    2017-07-01

    Full Text Available Ionic stress is one of the most important components of salinity and is brought about by excess Na+ accumulation, especially in the aerial parts of plants. Since Na+ interferes with K+ homeostasis, and especially given its involvement in numerous metabolic processes, maintaining a balanced cytosolic Na+/K+ ratio has become a key salinity tolerance mechanism. Achieving this homeostatic balance requires the activity of Na+ and K+ transporters and/or channels. The mechanism of Na+ and K+ uptake and translocation in glycophytes and halophytes is essentially the same, but glycophytes are more susceptible to ionic stress than halophytes. The transport mechanisms involve Na+ and/or K+ transporters and channels as well as non-selective cation channels. Thus, the question arises of whether the difference in salt tolerance between glycophytes and halophytes could be the result of differences in the proteins or in the expression of genes coding the transporters. The aim of this review is to seek answers to this question by examining the role of major Na+ and K+ transporters and channels in Na+ and K+ uptake, translocation and intracellular homeostasis in glycophytes. It turns out that these transporters and channels are equally important for the adaptation of glycophytes as they are for halophytes, but differential gene expression, structural differences in the proteins (single nucleotide substitutions, impacting affinity and post-translational modifications (phosphorylation account for the differences in their activity and hence the differences in tolerance between the two groups. Furthermore, lack of the ability to maintain stable plasma membrane (PM potentials following Na+-induced depolarization is also crucial for salt stress tolerance. This stable membrane potential is sustained by the activity of Na+/H+ antiporters such as SOS1 at the PM. Moreover, novel regulators of Na+ and K+ transport pathways including the Nax1 and Nax2 loci regulation of SOS1

  7. VPLIV GRADBENIH DEJAVNIKOV NA EKONOMIKO FOTONAPETOSTNIH SISTEMOV

    OpenAIRE

    Vrečič, Nuša

    2013-01-01

    Relavantne raziskave kažejo, da fotovoltaika postaja konkurenčna preostalim konvencionalnim virom, da bo v prihodnosti bistveno vplivala na proizvodnjo energije v Evropi ter pomembno vpliva na zamnjšanje emisij toplogrednih plinov. V teoretičnem delu so zajete tehnične zasnove in delovanje fotovoltaičnih elektrarn. Pri tehničnih zasnovah so opisani materiali in njihove lastnos