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{"title"=>"Screening of Dengue Virus Antiviral Activity of Marine Seaweeds by an In Situ Enzyme-Linked Immunosorbent Assay", "type"=>"journal", "authors"=>[{"first_name"=>"Andrea Cristine", "last_name"=>"Koishi", "scopus_author_id"=>"53063940200"}, {"first_name"=>"Paula Rodrigues", "last_name"=>"Zanello", "scopus_author_id"=>"55516714500"}, {"first_name"=>"Éverson Miguel", "last_name"=>"Bianco", "scopus_author_id"=>"26644785300"}, {"first_name"=>"Juliano", "last_name"=>"Bordignon", "scopus_author_id"=>"6602944005"}, {"first_name"=>"Claudia", "last_name"=>"Nunes Duarte dos Santos", "scopus_author_id"=>"55515874600"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84870798620", "sgr"=>"84870798620", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0051089", "pmid"=>"23227238", "isbn"=>"1932-6203", "pui"=>"366234424"}, "id"=>"63c46269-4603-3b5f-ae56-b43c9775c8e4", "abstract"=>"Dengue is a significant public health problem worldwide. Despite the important social and clinical impact, there is no vaccine or specific antiviral therapy for prevention and treatment of dengue virus (DENV) infection. Considering the above, drug discovery research for dengue is of utmost importance; in addition natural marine products provide diverse and novel chemical structures with potent biological activities that must be evaluated. In this study we propose a target-free approach for dengue drug discovery based on a novel, rapid, and economic in situ enzyme-linked immunosorbent assay and the screening of a panel of marine seaweed extracts. The in situ ELISA was standardized and validated for Huh7.5 cell line infected with all four serotypes of DENV, among them clinical isolates and a laboratory strain. Statistical analysis showed an average S/B of 7.2 and Z-factor of 0.62, demonstrating assay consistency and reliability. A panel of fifteen seaweed extracts was then screened at the maximum non-toxic dose previously determined by the MTT and Neutral Red cytotoxic assays. Eight seaweed extracts were able to reduce DENV infection of at least one serotype tested. Four extracts (Phaeophyta: Canistrocarpus cervicornis, Padina gymnospora; Rhodophyta: Palisada perforate; Chlorophyta: Caulerpa racemosa) were chosen for further evaluation, and time of addition studies point that they might act at an early stage of the viral infection cycle, such as binding or internalization.", "link"=>"http://www.mendeley.com/research/screening-dengue-virus-antiviral-activity-marine-seaweeds-situ-enzymelinked-immunosorbent-assay", "reader_count"=>73, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>4, "Librarian"=>1, "Student > Doctoral Student"=>2, "Researcher"=>12, "Student > Ph. D. Student"=>14, "Student > Postgraduate"=>4, "Other"=>5, "Student > Master"=>13, "Student > Bachelor"=>12, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>4, "Librarian"=>1, "Student > Doctoral Student"=>2, "Researcher"=>12, "Student > Ph. D. Student"=>14, "Student > Postgraduate"=>4, "Other"=>5, "Student > Master"=>13, "Student > Bachelor"=>12, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>5, "Agricultural and Biological Sciences"=>34, "Veterinary Science and Veterinary Medicine"=>1, "Chemistry"=>5, "Computer Science"=>2, "Economics, Econometrics and Finance"=>2, "Energy"=>1, "Environmental Science"=>2, "Biochemistry, Genetics and Molecular Biology"=>4, "Medicine and Dentistry"=>8, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Social Sciences"=>2, "Immunology and Microbiology"=>5, "Engineering"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>8}, "Social Sciences"=>{"Social Sciences"=>2}, "Unspecified"=>{"Unspecified"=>5}, "Environmental Science"=>{"Environmental Science"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "Engineering"=>{"Engineering"=>1}, "Chemistry"=>{"Chemistry"=>5}, "Energy"=>{"Energy"=>1}, "Economics, Econometrics and Finance"=>{"Economics, Econometrics and Finance"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>34}, "Computer Science"=>{"Computer Science"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Bangladesh"=>1, "Belgium"=>1, "Brazil"=>4, "Mexico"=>1, "United Kingdom"=>1, "French Polynesia"=>1}, "group_count"=>6}

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• http://doi.org/10.12980/APJTB.4.2014C1099
• http://doi.org/10.1007/s00253-014-5575-0
• http://doi.org/10.1038/srep05452
• http://doi.org/10.1074/jbc.M116.772897
• http://doi.org/10.1016/j.brainres.2013.10.002
• http://doi.org/10.3390/md16080265
• http://doi.org/10.3390/ijms19041093
• http://doi.org/10.1186/s12985-015-0443-9
• http://doi.org/10.3390/md14030052
• http://doi.org/10.1038/ncomms6268
• http://doi.org/10.1038/srep41864
• http://doi.org/10.1371/journal.pone.0112474
• http://doi.org/10.1099/acmi.0.000073
• http://doi.org/10.1186/s12985-016-0548-9
• http://doi.org/10.1016/j.fsi.2016.09.016

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/285922", "https://ndownloader.figshare.com/files/285971"], "description"=>"<div><p>Dengue is a significant public health problem worldwide. Despite the important social and clinical impact, there is no vaccine or specific antiviral therapy for prevention and treatment of dengue virus (DENV) infection. Considering the above, drug discovery research for dengue is of utmost importance; in addition natural marine products provide diverse and novel chemical structures with potent biological activities that must be evaluated. In this study we propose a target-free approach for dengue drug discovery based on a novel, rapid, and economic <em>in situ</em> enzyme-linked immunosorbent assay and the screening of a panel of marine seaweed extracts. The <em>in situ</em> ELISA was standardized and validated for Huh7.5 cell line infected with all four serotypes of DENV, among them clinical isolates and a laboratory strain. Statistical analysis showed an average S/B of 7.2 and Z-factor of 0.62, demonstrating assay consistency and reliability. A panel of fifteen seaweed extracts was then screened at the maximum non-toxic dose previously determined by the MTT and Neutral Red cytotoxic assays. Eight seaweed extracts were able to reduce DENV infection of at least one serotype tested. Four extracts (Phaeophyta: <em>Canistrocarpus cervicornis</em>, <em>Padina gymnospora</em>; Rhodophyta: <em>Palisada perforate</em>; Chlorophyta: <em>Caulerpa racemosa</em>) were chosen for further evaluation, and time of addition studies point that they might act at an early stage of the viral infection cycle, such as binding or internalization.</p> </div>", "links"=>[], "tags"=>["dengue", "antiviral", "seaweeds", "enzyme-linked", "immunosorbent", "assay"], "article_id"=>116361, "categories"=>["Cancer", "Biochemistry", "Pharmacology", "Immunology", "Biotechnology"], "users"=>["Andrea Cristine Koishi", "Paula Rodrigues Zanello", "Éverson Miguel Bianco", "Juliano Bordignon", "Claudia Nunes Duarte dos Santos"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0051089.s001", "https://dx.doi.org/10.1371/journal.pone.0051089.s002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Screening_of_Dengue_Virus_Antiviral_Activity_of_Marine_Seaweeds_by_an_In_Situ_Enzyme_Linked_Immunosorbent_Assay__/116361", "title"=>"Screening of Dengue Virus Antiviral Activity of Marine Seaweeds by an <em>In Situ</em> Enzyme-Linked Immunosorbent Assay", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-12-05 01:46:01"}
• {"files"=>["https://ndownloader.figshare.com/files/531107"], "description"=>"<p><i>In situ</i> ELISA assay workflow.</p>", "links"=>[], "tags"=>["elisa", "assay"], "article_id"=>201600, "categories"=>["Virology", "Biochemistry", "Infectious Diseases", "Immunology", "Pharmacology", "Biotechnology"], "users"=>["Andrea Cristine Koishi", "Paula Rodrigues Zanello", "Éverson Miguel Bianco", "Juliano Bordignon", "Claudia Nunes Duarte dos Santos"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0051089.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_situ_ELISA_assay_workflow_/201600", "title"=>"<i>In situ</i> ELISA assay workflow.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-05 00:26:40"}
• {"files"=>["https://ndownloader.figshare.com/files/531187"], "description"=>"<p>(<b>A</b>) Effect of different blocking buffers (skim milk and FBS) and fixatives (methanol:acetone and paraformaldehyde followed by triton X-100) for the assay. (<b>B</b>) Cells were seeded in a range of concentration and the influence for the assay performance was evaluated. (<b>C</b>) The ideal MOI for all DENV serotypes was evaluated 48 hours post-infection and (<b>D</b>) 72 hours post-infection. Values represent mean ± SD of three independent experiments performed in triplicate.</p>", "links"=>[], "tags"=>["elisa"], "article_id"=>201687, "categories"=>["Virology", "Biochemistry", "Infectious Diseases", "Immunology", "Pharmacology", "Biotechnology"], "users"=>["Andrea Cristine Koishi", "Paula Rodrigues Zanello", "Éverson Miguel Bianco", "Juliano Bordignon", "Claudia Nunes Duarte dos Santos"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0051089.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_situ_ELISA_optimization_/201687", "title"=>"<i>In situ</i> ELISA optimization.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-05 00:28:07"}
• {"files"=>["https://ndownloader.figshare.com/files/531295"], "description"=>"<p>(<b>A</b>) Correlation between the <i>in situ</i> ELISA and the foci-forming assay, where Huh7.5 was infected with DENV-1, DENV-2, DENV-3 and DENV-4 at a range of MOI (4–0.01). After 72 hours of infection the supernatant was used for the foci-forming assay and the cells submitted to the ELISA assay. Data from one representative experiment, analyzed using Pearson correlation test. (<b>B</b>) Comparison of the DRC and the IC50 for IFN-α 2A and DENV-4 infection, obtained with the <i>in situ</i> ELISA, foci-forming assay and a commercial NS1 antigen capture ELISA assay. Mean ± SD of three independent experiments, analyzed by sigmoidal dose-response curve (variable slope), the dashed line represents the detection limit of the foci-forming assay.</p>", "links"=>[], "tags"=>["elisa", "assay"], "article_id"=>201794, "categories"=>["Virology", "Biochemistry", "Infectious Diseases", "Immunology", "Pharmacology", "Biotechnology"], "users"=>["Andrea Cristine Koishi", "Paula Rodrigues Zanello", "Éverson Miguel Bianco", "Juliano Bordignon", "Claudia Nunes Duarte dos Santos"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0051089.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_situ_ELISA_assay_validation_/201794", "title"=>"<i>In situ</i> ELISA assay validation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-05 00:29:54"}
• {"files"=>["https://ndownloader.figshare.com/files/531368"], "description"=>"<p>(<b>A</b>) Huh7.5 were seeded in a 96-well plate and infected with DENV-1, (<b>B</b>) DENV-2 and (<b>C</b>) DENV-3 with an MOI of 4, and (<b>D</b>) DENV-4 with an MOI of 0.1. Interferon-α 2A (100 IU/ml) was used as a positive control and after 72 hours post-infection the ELISA was performed. Data were analyzed using one-way ANOVA followed by Tukey test. Values are mean ± SD of three independent experiments. *p<0.05.</p>", "links"=>[], "tags"=>["extracts", "antiviral"], "article_id"=>201861, "categories"=>["Virology", "Biochemistry", "Infectious Diseases", "Immunology", "Pharmacology", "Biotechnology"], "users"=>["Andrea Cristine Koishi", "Paula Rodrigues Zanello", "Éverson Miguel Bianco", "Juliano Bordignon", "Claudia Nunes Duarte dos Santos"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0051089.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Seaweed_extracts_antiviral_screening_/201861", "title"=>"Seaweed extracts antiviral screening.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-05 00:31:01"}
• {"files"=>["https://ndownloader.figshare.com/files/531470"], "description"=>"<p>Huh7.5 was seeded in a 96-well plate and infected with each strain with a MOI of 4 for 1 h30 min, then the seaweed extracts were added and after 72 hours the ELISA was performed. Data were analyzed using one-way ANOVA followed by Dunnett test. Values are mean ± SD of three independent experiments,*p<0.05.</p>", "links"=>[], "tags"=>["inhibition"], "article_id"=>201967, "categories"=>["Virology", "Biochemistry", "Infectious Diseases", "Immunology", "Pharmacology", "Biotechnology"], "users"=>["Andrea Cristine Koishi", "Paula Rodrigues Zanello", "Éverson Miguel Bianco", "Juliano Bordignon", "Claudia Nunes Duarte dos Santos"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0051089.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DENV_infection_inhibition_for_different_strains_serotypes_/201967", "title"=>"DENV infection inhibition for different strains/serotypes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-05 00:32:47"}
• {"files"=>["https://ndownloader.figshare.com/files/531569"], "description"=>"<p>(<b>A</b>) Huh 7.5 cells were infected with DENV-4 with an MOI of 0.1 following each treatment (before the infection, −1.5 h; during the infection, 0 h; and after the infection, +1.5 h) at the MNTD, the cells were submitted to the ELISA assay (<b>B</b>) and the supernatant for the foci-forming assay. The dashed line represents the detection limit of the foci-forming assay.</p>", "links"=>[], "tags"=>["studies", "seaweed", "extracts", "a8"], "article_id"=>202059, "categories"=>["Virology", "Biochemistry", "Infectious Diseases", "Immunology", "Pharmacology", "Biotechnology"], "users"=>["Andrea Cristine Koishi", "Paula Rodrigues Zanello", "Éverson Miguel Bianco", "Juliano Bordignon", "Claudia Nunes Duarte dos Santos"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0051089.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Time_of_addition_studies_with_seaweed_extracts_A1_A3_A8_and_A12_/202059", "title"=>"Time of addition studies with seaweed extracts A1, A3, A8 and A12.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-05 00:34:19"}
• {"files"=>["https://ndownloader.figshare.com/files/531624"], "description"=>"<p>(<b>A</b>) Huh 7.5 cells were infected with DENV-4 and treated during the infection in a range of concentration for each seaweed extract. (<b>B</b>) CC50, IC50 and SI were calculated. Mean ± SD of three independent experiments, analyzed by sigmoidal dose response curve (variable slope).</p>", "links"=>[], "tags"=>["seaweed", "extracts", "a8"], "article_id"=>202114, "categories"=>["Virology", "Biochemistry", "Infectious Diseases", "Immunology", "Pharmacology", "Biotechnology"], "users"=>["Andrea Cristine Koishi", "Paula Rodrigues Zanello", "Éverson Miguel Bianco", "Juliano Bordignon", "Claudia Nunes Duarte dos Santos"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0051089.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Dose_response_curve_for_the_seaweed_extracts_A1_A3_A8_and_A12_/202114", "title"=>"Dose response curve for the seaweed extracts A1, A3, A8 and A12.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-05 00:35:14"}
• {"files"=>["https://ndownloader.figshare.com/files/531780"], "description"=>"<p>(<b>A</b>) Binding inhibition assay. (<b>B</b>) Internalization inhibition assay. Data were analyzed using one-way ANOVA followed by Dunnett test. Values are mean ± SD of three independent experiments,*p<0.05.</p>", "links"=>[], "tags"=>["seaweed", "extracts", "a8", "a12", "denv-4"], "article_id"=>202270, "categories"=>["Virology", "Biochemistry", "Infectious Diseases", "Immunology", "Pharmacology", "Biotechnology"], "users"=>["Andrea Cristine Koishi", "Paula Rodrigues Zanello", "Éverson Miguel Bianco", "Juliano Bordignon", "Claudia Nunes Duarte dos Santos"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0051089.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_the_seaweed_extracts_A1_A3_A8_and_A12_on_DENV_4_entry_process_/202270", "title"=>"Effect of the seaweed extracts A1, A3, A8 and A12 on DENV-4 entry process.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-12-05 00:37:50"}
• {"files"=>["https://ndownloader.figshare.com/files/531836"], "description"=>"a<p>CC50∶50% cytotoxic concentration (µg/ml).</p>b<p>MNTD: maximum non- toxic dose based on MTT and Neutral Red assays.</p>c<p>Phaeophyta = Brown seaweeds; Rhodophyta = Red seaweeds; Chlorophyta = Green seaweeds.</p>", "links"=>[], "tags"=>["seaweeds", "antiviral", "cytotoxicity"], "article_id"=>202331, "categories"=>["Virology", "Biochemistry", "Infectious Diseases", "Immunology", "Pharmacology", "Biotechnology"], "users"=>["Andrea Cristine Koishi", "Paula Rodrigues Zanello", "Éverson Miguel Bianco", "Juliano Bordignon", "Claudia Nunes Duarte dos Santos"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0051089.t001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Marine_seaweeds_selected_for_antiviral_screening_location_of_collection_and_cytotoxicity_activity_in_Huh7_5_cells_/202331", "title"=>"Marine seaweeds selected for antiviral screening, location of collection and cytotoxicity activity in Huh7.5 cells.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-12-05 00:38:51"}

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Relative Metric

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F1000Prime