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{"title"=>"Role of permissive neuraminidase mutations in influenza A/Brisbane/59/2007-like (H1N1) viruses", "type"=>"journal", "authors"=>[{"first_name"=>"Yacine", "last_name"=>"Abed", "scopus_author_id"=>"7005143651"}, {"first_name"=>"Andrés", "last_name"=>"Pizzorno", "scopus_author_id"=>"37079431400"}, {"first_name"=>"Xavier", "last_name"=>"Bouhy", "scopus_author_id"=>"35268141400"}, {"first_name"=>"Guy", "last_name"=>"Boivin", "scopus_author_id"=>"24309591300"}], "year"=>2011, "source"=>"PLoS Pathogens", "identifiers"=>{"isbn"=>"1553-7374 (Electronic)\\r1553-7366 (Linking)", "sgr"=>"84855259703", "pui"=>"364024586", "pmid"=>"22174688", "issn"=>"15537366", "scopus"=>"2-s2.0-84855259703", "doi"=>"10.1371/journal.ppat.1002431"}, "id"=>"8ff21a21-3c7d-378b-bbb3-e3780691ce55", "abstract"=>"Neuraminidase (NA) mutations conferring resistance to NA inhibitors were believed to compromise influenza virus fitness. Unexpectedly, an oseltamivir-resistant A/Brisbane/59/2007 (Bris07)-like H1N1 H275Y NA variant emerged in 2007 and completely replaced the wild-type (WT) strain in 2008-2009. The NA of such variant contained additional NA changes (R222Q, V234M and D344N) that potentially counteracted the detrimental effect of the H275Y mutation on viral fitness. Here, we rescued a recombinant Bris07-like WT virus and 4 NA mutants/revertants (H275Y, H275Y/Q222R, H275Y/M234V and H275Y/N344D) and characterized them in vitro and in ferrets. A fluorometric-based NA assay was used to determine Vmax and Km values. Replicative capacities were evaluated by yield assays in ST6Gal1-MDCK cells. Recombinant NA proteins were expressed in 293T cells and surface NA activity was determined. Infectivity and contact transmission experiments were evaluated for the WT, H275Y and H275Y/Q222R recombinants in ferrets. The H275Y mutation did not significantly alter Km and Vmax values compared to WT. The H275Y/N344D mutant had a reduced affinity (Km of 50 vs 12 µM) whereas the H275Y/M234V mutant had a reduced activity (22 vs 28 U/sec). In contrast, the H275Y/Q222R mutant showed a significant decrease of both affinity (40 µM) and activity (7 U/sec). The WT, H275Y, H275Y/M234V and H275Y/N344D recombinants had comparable replicative capacities contrasting with H275Y/Q222R mutant whose viral titers were significantly reduced. All studied mutations reduced the cell surface NA activity compared to WT with the maximum reduction being obtained for the H275Y/Q222R mutant. Comparable infectivity and transmissibility were seen between the WT and the H275Y mutant in ferrets whereas the H275Y/Q222R mutant was associated with significantly lower lung viral titers. In conclusion, the Q222R reversion mutation compromised Bris07-like H1N1 virus in vitro and in vivo. Thus, the R222Q NA mutation present in the WT virus may have facilitated the emergence of NAI-resistant Bris07 variants.", "link"=>"http://www.mendeley.com/research/role-permissive-neuraminidase-mutations-influenza-abrisbane592007like-h1n1-viruses", "reader_count"=>33, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Student > Doctoral Student"=>4, "Researcher"=>7, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>2, "Other"=>2, "Student > Master"=>4, "Student > Bachelor"=>3, "Lecturer"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Student > Doctoral Student"=>4, "Researcher"=>7, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>2, "Other"=>2, "Student > Master"=>4, "Student > Bachelor"=>3, "Lecturer"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>4, "Mathematics"=>1, "Agricultural and Biological Sciences"=>12, "Medicine and Dentistry"=>4, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Chemistry"=>1, "Social Sciences"=>1, "Computer Science"=>1, "Immunology and Microbiology"=>5}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Chemistry"=>{"Chemistry"=>1}, "Social Sciences"=>{"Social Sciences"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>12}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Mathematics"=>{"Mathematics"=>1}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Australia"=>1}, "group_count"=>3}

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/704710"], "description"=>"<p>Confluent ST6Gal1-MDCK cells were infected with recombinant viruses at a multiplicity of infection (MOI) of 0.001 PFU/cell. Supernatants were harvested at 12 h, 24 h, 36 h, 48 h and 60 h post-infection and titrated by standard plaque assays. The mean values for three experiments with standard deviations are presented. *<i>P</i><0.05 and ***<i>P</i><0.001 for differences in viral titers when compared to the recombinant WT virus.</p>", "links"=>[], "tags"=>["kinetics", "recombinant", "viruses"], "article_id"=>375067, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Yacine Abed", "Andrés Pizzorno", "Xavier Bouhy", "Guy Boivin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002431.g003", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Replication_kinetics_of_recombinant_A_Brisbane_59_2007_like_viruses_in_vitro_/375067", "title"=>"Replication kinetics of recombinant A/Brisbane/59/2007-like viruses <i>in vitro</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 10:58:14"}
• {"files"=>["https://ndownloader.figshare.com/files/704655"], "description"=>"<p>293T cells were transfected with pCAGGS-PA, -PB1, -PB2, and -NP plasmids in addition to plasmids expressing the WT or mutant A/Brisbane/59/2007-like neuraminidases (NA) proteins. At 24 h post-transfection, cells were treated with a non-lysing buffer and surface NA activity was measured by using the fluorogenic substrate (MUNANA). Percent surface NA activities were determined in triplicate experiments ± standard deviations. **<i>P</i><0.01 and ***<i>P</i><0.001 compared to the WT surface NA activity.</p>", "links"=>[], "tags"=>["recombinant", "neuraminidase"], "article_id"=>375020, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Yacine Abed", "Andrés Pizzorno", "Xavier Bouhy", "Guy Boivin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002431.g002", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Surface_activity_of_recombinant_A_Brisbane_59_2007_like_H1N1_neuraminidase_proteins_/375020", "title"=>"Surface activity of recombinant A/Brisbane/59/2007-like (H1N1) neuraminidase proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 10:57:58"}
• {"files"=>["https://ndownloader.figshare.com/files/704942"], "description"=>"a<p>Values indicate mean <i>Km</i> and relative NA activity (<i>Vmax)</i> values of a representative experiment performed in triplicate ± standard deviations (SD).</p>b<p>Values indicate mean plaque area (N = 16) ± SD.</p><p>*P<0.05,</p><p>***P<0.001 compared to WT.</p>", "links"=>[], "tags"=>["enzymatic", "properties", "plaque", "areas", "recombinant"], "article_id"=>375305, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Yacine Abed", "Andrés Pizzorno", "Xavier Bouhy", "Guy Boivin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002431.t001", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Neuraminidase_enzymatic_properties_and_plaque_areas_of_recombinant_A_Brisbane_59_2007_like_H1N1_viruses_/375305", "title"=>"Neuraminidase enzymatic properties and plaque areas of recombinant A/Brisbane/59/2007-like (H1N1) viruses.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 10:59:28"}
• {"files"=>["https://ndownloader.figshare.com/files/704983"], "description"=>"a<p>Values indicated mean IC₅₀ values of three experiments ± standard deviations.</p><p>**<i>P<0.01</i> compared to the single H275Y mutant.</p>", "links"=>[], "tags"=>["profiles", "recombinant", "viruses", "neuraminidase", "inhibitors", "assessed", "munana", "na", "inhibition"], "article_id"=>375336, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Yacine Abed", "Andrés Pizzorno", "Xavier Bouhy", "Guy Boivin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002431.t002", "stats"=>{"downloads"=>4, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Susceptibility_profiles_of_recombinant_A_Brisbane_59_2007_like_H1N1_viruses_against_neuraminidase_NA_inhibitors_as_assessed_by_MUNANA_NA_inhibition_assays_/375336", "title"=>"Susceptibility profiles of recombinant A/Brisbane/59/2007-like (H1N1) viruses against neuraminidase (NA) inhibitors as assessed by MUNANA NA inhibition assays.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 10:59:39"}
• {"files"=>["https://ndownloader.figshare.com/files/704777"], "description"=>"<p>Body temperatures were recorded by rectal thermometer during 10 days post-inoculation in groups of 4 index ferrets infected with 1.25×10⁵ PFU of recombinant A/Brisbane/59/2007-like wild-type (WT) virus as well as H275Y and H275Y/Q222R mutants (A) and in groups of 4 naïve ferrets that were placed in direct contact with index ferrets 24 h later (B).</p>", "links"=>[], "tags"=>["temperatures", "infected"], "article_id"=>375139, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Yacine Abed", "Andrés Pizzorno", "Xavier Bouhy", "Guy Boivin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002431.g004", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Body_temperatures_of_infected_and_contact_ferrets_/375139", "title"=>"Body temperatures of infected and contact ferrets.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 10:58:36"}
• {"files"=>["https://ndownloader.figshare.com/files/704853"], "description"=>"<p>Mean viral titers ± standard deviations were determined in nasal washes by using standard plaque assays in groups of 4 index ferrets infected with 1.25×10⁵ PFU of recombinant A/Brisbane/59/2007-like wild-type (WT) virus as well as H275Y and H275Y/Q222R mutants (A) and in groups of 4 naïve ferrets that were placed in direct contact with index ferrets 24 h later (B).*<i>P</i><0.05 and **<i>P</i><0.01 for differences in viral titers when compared to the recombinant WT virus.</p>", "links"=>[], "tags"=>["viral", "titers", "nasal", "samples", "infected"], "article_id"=>375210, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Yacine Abed", "Andrés Pizzorno", "Xavier Bouhy", "Guy Boivin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002431.g005", "stats"=>{"downloads"=>3, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mean_viral_titers_in_nasal_wash_samples_of_infected_and_contact_ferrets_/375210", "title"=>"Mean viral titers in nasal wash samples of infected and contact ferrets.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 10:59:01"}
• {"files"=>["https://ndownloader.figshare.com/files/704597"], "description"=>"<p>The rate of substrate conversion velocity (V₀) by NA enzymes from a standardized dose of 10⁶ PFU/ml of recombinant virus was determined. The fluorogenic substrate (MUNANA) was used at final concentrations of 0 to 3000 µM. Fluorescence was measured every 90 sec for 53 min at 37 °C using excitation and emission wavelengths of 355 and 460 nm, respectively. The data of one representative experiment performed in triplicate is shown.</p>", "links"=>[], "tags"=>["enzyme", "kinetics", "recombinant"], "article_id"=>374956, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Yacine Abed", "Andrés Pizzorno", "Xavier Bouhy", "Guy Boivin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002431.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Neuraminidase_NA_enzyme_kinetics_of_recombinant_A_Brisbane_59_2007_like_H1N1_viruses_/374956", "title"=>"Neuraminidase (NA) enzyme kinetics of recombinant A/Brisbane/59/2007-like (H1N1) viruses.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 10:57:38"}

PMC Usage Stats | Further Information

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Relative Metric

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F1000Prime | Further Information

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• {"year"=>2014, "month"=>8, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2014-08-25T08:33:46Z"}
• {"year"=>2014, "month"=>9, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2014-09-29T08:33:47Z"}
• {"year"=>2014, "month"=>10, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2014-10-27T08:33:39Z"}
• {"year"=>2014, "month"=>11, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2014-11-24T09:32:24Z"}
• {"year"=>2014, "month"=>12, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2014-12-29T09:42:09Z"}
• {"year"=>2015, "month"=>1, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2015-01-26T09:40:46Z"}
• {"year"=>2015, "month"=>2, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2015-02-23T09:40:48Z"}
• {"year"=>2015, "month"=>3, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2015-03-23T08:40:59Z"}
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• {"year"=>2015, "month"=>5, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2015-05-25T08:40:49Z"}
• {"year"=>2015, "month"=>6, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2015-06-29T08:40:50Z"}
• {"year"=>2015, "month"=>7, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2015-07-27T08:40:41Z"}
• {"year"=>2015, "month"=>8, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2015-08-31T08:40:52Z"}
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• {"year"=>2015, "month"=>10, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2015-10-26T08:40:55Z"}
• {"year"=>2015, "month"=>11, "doi"=>"10.1371/journal.ppat.1002431", "f1000_id"=>"13489134", "url"=>"http://f1000.com/prime/13489134", "score"=>2, "classifications"=>["confirmation", "new_finding"], "updated_at"=>"2015-11-30T09:40:56Z"}
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