Enzymatic Properties of the Neuraminidase of Seasonal H1N1 Influenza Viruses Provide Insights for the Emergence of Natural Resistance to Oseltamivir
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  • {"files"=>["https://ndownloader.figshare.com/files/925750"], "description"=>"<p><i>p-</i>Values are the result of the Student's t test between the mean of K<sub>m</sub>, V<sub>m</sub>, or K<sub>i</sub> of the sensitive viruses versus either the resistant ones or the pre-2007–2008 viruses. Student's t test was also performed between the mean K<sub>m</sub> of the sensitive viruses versus the mean K<sub>m</sub> of the early seasonal virus #0006/07.</p><p>Amino acids that differ from the NC99 strain are shown in bold.</p>a<p>Except for the reference vaccine strains A/New Caledonia/20/99(H1N1) (NC99) and A/Solomon Islands//2006(H1N1) (SI06), all other viruses were A(H1N1) clinical isolates from the NIC (Northern-France) named by order number and year of isolation. Virus 0692/07 is an isolate from the 2006–2007 season.</p>b<p>Week of sampling of specimen during the 2007–2008 season.</p>c<p>IC50 determined essentially as described in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000103#ppat.1000103-RameixWelti1\" target=\"_blank\">[20]</a>,<a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000103#ppat.1000103-Potier1\" target=\"_blank\">[30]</a> using the MUNANA substrate at a final concentration of 100 μM.</p>d<p>The Michaelis-Menten constant K<sub>m</sub> was determined by enzymatic kinetic analyses performed on inactivated virus suspensions. Briefly, kinetics were performed using MUNANA concentrations ranging from 5 to 100 μM. Initial velocity of the reaction was calculated and plotted as a function of the MUNANA concentration. K<sub>m</sub> and V<sub>m</sub> were calculated using a nonlinear regression of the curve according to the Michaelis-Menten equation. V<sub>m</sub> values were expressed in arbitrary units (U/sec).</p>e<p>Hemagglutination (HA) titers were determined in duplicate by standard procedures using guinea pig red blood cells. Titers are expressed as the mean of the reciprocal of the last virus dilutions showing hemagglutination.</p>f<p>K<sub>i</sub> determinations rely on enzymatic kinetic analyses. The kinetics were performed in the presence of variable concentrations of inhibitor (0 to 2,000 nM) and a constant MUNANA concentration (20 μM). Calculation of the K<sub>i</sub> was performed by nonlinear regression of the plot of the initial velocity as a function of the concentration of inhibitor.</p>g<p>K<sub>m</sub> and V<sub>m</sub> values are given as the mean of 2, 3<sup>*</sup>, 4<sup>**</sup>, or 7<sup>***</sup> determinations.</p>h<p>Values correspond to the mean ± standard deviation of the mean K<sub>m</sub> or V<sub>m</sub> values of the 5 pre-2007–2008 viruses , the 6 sensitive viruses, and the 11 resistant viruses. Means and <i>p</i>-values apply to virus groups indicated by —.</p>i<p>K<sub>i</sub> values were determined once except for the SI06 reference strain, for which three independent determinations were performed, and the 0497/07 and 644/07 viruses, for which two independent determinations were performed.</p>j<p>Values correspond to the mean ± standard deviation of the mean K<sub>i</sub> values of the 5 pre-2007–2008 viruses, the 6 sensitive viruses, and the 11 resistant viruses. Means and <i>p</i>-values apply to virus groups indicated by —.</p><p>ND, not determined; NS not significant.</p>", "links"=>[], "tags"=>["neuraminidase", "h1n1", "viruses"], "article_id"=>596200, "categories"=>["Virology", "Infectious Diseases"], "users"=>["Marie-Anne Rameix-Welti", "Vincent Enouf", "Frédérique Cuvelier", "Patricia Jeannin", "Sylvie van der Werf"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000103.t001", "stats"=>{"downloads"=>3, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Properties_of_the_Neuraminidase_of_H1N1_Viruses_from_the_2007_8211_2008_Season_/596200", "title"=>"Properties of the Neuraminidase of H1N1 Viruses from the 2007–2008 Season", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-21 09:55:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/925515"], "description"=>"<p>Two sensitive (A/Paris/497/2007 and A/Paris/1149/2008) and two resistant (A/Paris/644/2007 and A/Paris/1170/2008) viruses from the 2007–2008 influenza season, as well as the reference strain A/Solomon Islands/3/2006 and an isolate from the 2003–2004 season (A/Paris/650/2004), were amplified and titrated on MDCK cells. The indicated viruses were then used to infect MDCK SIAT-1 cells <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000103#ppat.1000103-Matrosovich1\" target=\"_blank\">[31]</a> at an m.o.i. of 0.001 and incubated for 72 hours at 35°C in the presence of 1 μg/ml TPCK trypsin. At the indicated time points, the supernatants were harvested and virus titers were determined by plaque assays on MDCK cells.</p>", "links"=>[], "tags"=>["kinetics", "h1n1", "viruses", "resistant"], "article_id"=>595961, "categories"=>["Virology", "Infectious Diseases"], "users"=>["Marie-Anne Rameix-Welti", "Vincent Enouf", "Frédérique Cuvelier", "Patricia Jeannin", "Sylvie van der Werf"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000103.g001", "stats"=>{"downloads"=>3, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Growth_Kinetics_of_H1N1_Viruses_from_the_2007_8211_2008_Season_Sensitive_or_Resistant_to_Oseltamivir_/595961", "title"=>"Growth Kinetics of H1N1 Viruses from the 2007–2008 Season Sensitive or Resistant to Oseltamivir.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:53:52"}
  • {"files"=>["https://ndownloader.figshare.com/files/925791"], "description"=>"<p>ND, not determined.</p>", "links"=>[], "tags"=>["virology/virus evolution and symbiosis", "virology/emerging viral diseases", "virology/antivirals, including modes of action and resistance"], "article_id"=>596241, "categories"=>["Virology", "Infectious Diseases"], "users"=>["Marie-Anne Rameix-Welti", "Vincent Enouf", "Frédérique Cuvelier", "Patricia Jeannin", "Sylvie van der Werf"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000103.t002", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Accession_Numbers_/596241", "title"=>"Accession Numbers", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-21 09:55:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/925585"], "description"=>"<p>The phylogenetic analysis was performed on the alignment of sequences from nucleotides 87 to 995 (H1) or 90 to 1286 (N1) (numbering from ATG). The dendrogram was constructed by genetic distance matrix and calculated with the DNADIST program using the Kimura-2 parameters model with transition-to-transversion ratio of 2.0 and neighbor-joining analysis in the PHYLIP package <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000103#ppat.1000103-Felsenstein1\" target=\"_blank\">[32]</a>,<a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000103#ppat.1000103-Felsenstein2\" target=\"_blank\">[33]</a>. Sequences from the H1 or the N1 from A/NewCaledonia/20/99 were used as outgroup. Bootstrap values of 1,000 replicas are given as percentages at the nodes. Isolates from Northern-France from the 2006–2007 season are in italics. Viruses with a Y275 in the neuraminidase sequence are shown in bold and vaccine strains in capitals. Published sequences were issued from the influenza sequence database at Los Alamos National Laboratory <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000103#ppat.1000103-Macken1\" target=\"_blank\">[22]</a>.</p>", "links"=>[], "tags"=>["relationships", "h1", "n1"], "article_id"=>596033, "categories"=>["Virology", "Infectious Diseases"], "users"=>["Marie-Anne Rameix-Welti", "Vincent Enouf", "Frédérique Cuvelier", "Patricia Jeannin", "Sylvie van der Werf"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000103.g002", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phylogenetic_Relationships_of_the_H1_and_N1_Genes_/596033", "title"=>"Phylogenetic Relationships of the H1 and N1 Genes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:54:20"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"8", "full-text"=>"4", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"3", "year"=>"2015", "month"=>"11"}
  • {"unique-ip"=>"7", "full-text"=>"7", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"12"}
  • {"unique-ip"=>"3", "full-text"=>"5", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"1"}
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  • {"unique-ip"=>"5", "full-text"=>"3", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"10"}
  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"11"}
  • {"unique-ip"=>"1", "full-text"=>"1", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"12"}
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  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"9"}
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  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"3"}
  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"1"}
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  • {"unique-ip"=>"5", "full-text"=>"2", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"7"}
  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"8"}
  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"10"}
  • {"unique-ip"=>"10", "full-text"=>"9", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
  • {"unique-ip"=>"10", "full-text"=>"11", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"4", "full-text"=>"4", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"7", "full-text"=>"7", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"4", "full-text"=>"3", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
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Relative Metric

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