Withaferin-A Reduces Type I Collagen Expression In Vitro and Inhibits Development of Myocardial Fibrosis In Vivo
Events
Loading … Spinner

Mendeley | Further Information

{"title"=>"Withaferin-A reduces type I collagen expression in vitro and inhibits development of myocardial fibrosis in vivo", "type"=>"journal", "authors"=>[{"first_name"=>"Azariyas A.", "last_name"=>"Challa", "scopus_author_id"=>"54082784800"}, {"first_name"=>"Milica", "last_name"=>"Vukmirovic", "scopus_author_id"=>"55337133000"}, {"first_name"=>"John", "last_name"=>"Blackmon", "scopus_author_id"=>"57197199392"}, {"first_name"=>"Branko", "last_name"=>"Stefanovic", "scopus_author_id"=>"7006143855"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"22900077", "sgr"=>"84865011936", "scopus"=>"2-s2.0-84865011936", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0042989", "pui"=>"365442003", "issn"=>"19326203"}, "id"=>"5f49f4dc-cfe9-317a-ae69-e98ad06251c8", "abstract"=>"Type I collagen is the most abundant protein in the human body. Its excessive synthesis results in fibrosis of various organs. Fibrosis is a major medical problem without an existing cure. Excessive synthesis of type I collagen in fibrosis is primarily due to stabilization of collagen mRNAs. We recently reported that intermediate filaments composed of vimentin regulate collagen synthesis by stabilizing collagen mRNAs. Vimentin is a primary target of Withaferin-A (WF-A). Therefore, we hypothesized that WF-A may reduce type I collagen production by disrupting vimentin filaments and decreasing the stability of collagen mRNAs. This study is to determine if WF-A exhibits anti-fibrotic properties in vitro and in vivo and to elucidate the molecular mechanisms of its action. In lung, skin and heart fibroblasts WF-A disrupted vimentin filaments at concentrations of 0.5-1.5 µM and reduced 3 fold the half-lives of collagen α1(I) and α2(I) mRNAs and protein expression. In addition, WF-A inhibited TGF-β1 induced phosphorylation of TGF-β1 receptor I, Smad3 phosphorylation and transcription of collagen genes. WF-A also inhibited in vitro activation of primary hepatic stellate cells and decreased their type I collagen expression. In mice, administration of 4 mg/kg WF-A daily for 2 weeks reduced isoproterenol-induced myocardial fibrosis by 50%. Our findings provide strong evidence that Withaferin-A could act as an anti-fibrotic compound against fibroproliferative diseases, including, but not limited to, cardiac interstitial fibrosis.", "link"=>"http://www.mendeley.com/research/withaferina-reduces-type-i-collagen-expression-vitro-inhibits-development-myocardial-fibrosis-vivo", "reader_count"=>23, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>2, "Researcher"=>6, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>5, "Student > Master"=>3, "Other"=>2, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>2, "Researcher"=>6, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>5, "Student > Master"=>3, "Other"=>2, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>3, "Medicine and Dentistry"=>7, "Agricultural and Biological Sciences"=>8, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Physics and Astronomy"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>7}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>8}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "group_count"=>0}

Scopus | Further Information

{"@_fa"=>"true", "link"=>[{"@_fa"=>"true", "@ref"=>"self", "@href"=>"https://api.elsevier.com/content/abstract/scopus_id/84865011936"}, {"@_fa"=>"true", "@ref"=>"author-affiliation", "@href"=>"https://api.elsevier.com/content/abstract/scopus_id/84865011936?field=author,affiliation"}, {"@_fa"=>"true", "@ref"=>"scopus", "@href"=>"https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84865011936&origin=inward"}, {"@_fa"=>"true", "@ref"=>"scopus-citedby", "@href"=>"https://www.scopus.com/inward/citedby.uri?partnerID=HzOxMe3b&scp=84865011936&origin=inward"}], "prism:url"=>"https://api.elsevier.com/content/abstract/scopus_id/84865011936", "dc:identifier"=>"SCOPUS_ID:84865011936", "eid"=>"2-s2.0-84865011936", "dc:title"=>"Withaferin-A reduces type I collagen expression in vitro and inhibits development of myocardial fibrosis in vivo", "dc:creator"=>"Challa A.", "prism:publicationName"=>"PLoS ONE", "prism:eIssn"=>"19326203", "prism:volume"=>"7", "prism:issueIdentifier"=>"8", "prism:pageRange"=>nil, "prism:coverDate"=>"2012-08-10", "prism:coverDisplayDate"=>"10 August 2012", "prism:doi"=>"10.1371/journal.pone.0042989", "citedby-count"=>"27", "affiliation"=>[{"@_fa"=>"true", "affilname"=>"Florida State University", "affiliation-city"=>"Tallahassee", "affiliation-country"=>"United States"}], "pubmed-id"=>"22900077", "prism:aggregationType"=>"Journal", "subtype"=>"ar", "subtypeDescription"=>"Article", "article-number"=>"e42989", "source-id"=>"10600153309", "openaccess"=>"1", "openaccessFlag"=>true}

Facebook

  • {"url"=>"http%3A%2F%2Fjournals.plos.org%2Fplosone%2Farticle%3Fid%3D10.1371%252Fjournal.pone.0042989", "share_count"=>0, "like_count"=>0, "comment_count"=>0, "click_count"=>0, "total_count"=>0}

Counter

  • {"month"=>"8", "year"=>"2012", "pdf_views"=>"49", "xml_views"=>"2", "html_views"=>"254"}
  • {"month"=>"9", "year"=>"2012", "pdf_views"=>"18", "xml_views"=>"0", "html_views"=>"52"}
  • {"month"=>"10", "year"=>"2012", "pdf_views"=>"14", "xml_views"=>"0", "html_views"=>"53"}
  • {"month"=>"11", "year"=>"2012", "pdf_views"=>"15", "xml_views"=>"1", "html_views"=>"52"}
  • {"month"=>"12", "year"=>"2012", "pdf_views"=>"6", "xml_views"=>"0", "html_views"=>"39"}
  • {"month"=>"1", "year"=>"2013", "pdf_views"=>"17", "xml_views"=>"0", "html_views"=>"63"}
  • {"month"=>"2", "year"=>"2013", "pdf_views"=>"14", "xml_views"=>"1", "html_views"=>"48"}
  • {"month"=>"3", "year"=>"2013", "pdf_views"=>"10", "xml_views"=>"0", "html_views"=>"61"}
  • {"month"=>"4", "year"=>"2013", "pdf_views"=>"12", "xml_views"=>"1", "html_views"=>"58"}
  • {"month"=>"5", "year"=>"2013", "pdf_views"=>"22", "xml_views"=>"1", "html_views"=>"74"}
  • {"month"=>"6", "year"=>"2013", "pdf_views"=>"23", "xml_views"=>"0", "html_views"=>"91"}
  • {"month"=>"7", "year"=>"2013", "pdf_views"=>"16", "xml_views"=>"0", "html_views"=>"80"}
  • {"month"=>"8", "year"=>"2013", "pdf_views"=>"11", "xml_views"=>"0", "html_views"=>"44"}
  • {"month"=>"9", "year"=>"2013", "pdf_views"=>"9", "xml_views"=>"0", "html_views"=>"48"}
  • {"month"=>"10", "year"=>"2013", "pdf_views"=>"12", "xml_views"=>"3", "html_views"=>"91"}
  • {"month"=>"11", "year"=>"2013", "pdf_views"=>"6", "xml_views"=>"1", "html_views"=>"58"}
  • {"month"=>"12", "year"=>"2013", "pdf_views"=>"14", "xml_views"=>"2", "html_views"=>"89"}
  • {"month"=>"1", "year"=>"2014", "pdf_views"=>"5", "xml_views"=>"0", "html_views"=>"22"}
  • {"month"=>"2", "year"=>"2014", "pdf_views"=>"7", "xml_views"=>"0", "html_views"=>"50"}
  • {"month"=>"3", "year"=>"2014", "pdf_views"=>"6", "xml_views"=>"2", "html_views"=>"62"}
  • {"month"=>"4", "year"=>"2014", "pdf_views"=>"9", "xml_views"=>"1", "html_views"=>"58"}
  • {"month"=>"5", "year"=>"2014", "pdf_views"=>"13", "xml_views"=>"0", "html_views"=>"101"}
  • {"month"=>"6", "year"=>"2014", "pdf_views"=>"5", "xml_views"=>"0", "html_views"=>"77"}
  • {"month"=>"7", "year"=>"2014", "pdf_views"=>"6", "xml_views"=>"0", "html_views"=>"56"}
  • {"month"=>"8", "year"=>"2014", "pdf_views"=>"7", "xml_views"=>"2", "html_views"=>"30"}
  • {"month"=>"9", "year"=>"2014", "pdf_views"=>"14", "xml_views"=>"2", "html_views"=>"39"}
  • {"month"=>"10", "year"=>"2014", "pdf_views"=>"8", "xml_views"=>"1", "html_views"=>"32"}
  • {"month"=>"11", "year"=>"2014", "pdf_views"=>"3", "xml_views"=>"3", "html_views"=>"26"}
  • {"month"=>"12", "year"=>"2014", "pdf_views"=>"7", "xml_views"=>"1", "html_views"=>"34"}
  • {"month"=>"1", "year"=>"2015", "pdf_views"=>"2", "xml_views"=>"0", "html_views"=>"44"}
  • {"month"=>"2", "year"=>"2015", "pdf_views"=>"3", "xml_views"=>"0", "html_views"=>"24"}
  • {"month"=>"3", "year"=>"2015", "pdf_views"=>"1", "xml_views"=>"0", "html_views"=>"38"}
  • {"month"=>"4", "year"=>"2015", "pdf_views"=>"4", "xml_views"=>"1", "html_views"=>"24"}
  • {"month"=>"5", "year"=>"2015", "pdf_views"=>"9", "xml_views"=>"0", "html_views"=>"48"}
  • {"month"=>"6", "year"=>"2015", "pdf_views"=>"7", "xml_views"=>"0", "html_views"=>"27"}
  • {"month"=>"7", "year"=>"2015", "pdf_views"=>"1", "xml_views"=>"0", "html_views"=>"22"}
  • {"month"=>"8", "year"=>"2015", "pdf_views"=>"2", "xml_views"=>"1", "html_views"=>"22"}
  • {"month"=>"9", "year"=>"2015", "pdf_views"=>"4", "xml_views"=>"0", "html_views"=>"29"}
  • {"month"=>"10", "year"=>"2015", "pdf_views"=>"11", "xml_views"=>"0", "html_views"=>"31"}
  • {"month"=>"11", "year"=>"2015", "pdf_views"=>"1", "xml_views"=>"0", "html_views"=>"27"}
  • {"month"=>"12", "year"=>"2015", "pdf_views"=>"4", "xml_views"=>"0", "html_views"=>"28"}
  • {"month"=>"1", "year"=>"2016", "pdf_views"=>"5", "xml_views"=>"1", "html_views"=>"28"}
  • {"month"=>"2", "year"=>"2016", "pdf_views"=>"0", "xml_views"=>"0", "html_views"=>"28"}
  • {"month"=>"3", "year"=>"2016", "pdf_views"=>"7", "xml_views"=>"0", "html_views"=>"38"}
  • {"month"=>"4", "year"=>"2016", "pdf_views"=>"4", "xml_views"=>"0", "html_views"=>"25"}
  • {"month"=>"5", "year"=>"2016", "pdf_views"=>"5", "xml_views"=>"0", "html_views"=>"18"}
  • {"month"=>"6", "year"=>"2016", "pdf_views"=>"7", "xml_views"=>"0", "html_views"=>"20"}
  • {"month"=>"7", "year"=>"2016", "pdf_views"=>"6", "xml_views"=>"0", "html_views"=>"39"}
  • {"month"=>"8", "year"=>"2016", "pdf_views"=>"4", "xml_views"=>"0", "html_views"=>"13"}
  • {"month"=>"9", "year"=>"2016", "pdf_views"=>"13", "xml_views"=>"0", "html_views"=>"36"}
  • {"month"=>"10", "year"=>"2016", "pdf_views"=>"8", "xml_views"=>"0", "html_views"=>"26"}
  • {"month"=>"11", "year"=>"2016", "pdf_views"=>"2", "xml_views"=>"0", "html_views"=>"16"}
  • {"month"=>"12", "year"=>"2016", "pdf_views"=>"5", "xml_views"=>"0", "html_views"=>"29"}
  • {"month"=>"1", "year"=>"2017", "pdf_views"=>"9", "xml_views"=>"0", "html_views"=>"27"}
  • {"month"=>"2", "year"=>"2017", "pdf_views"=>"5", "xml_views"=>"0", "html_views"=>"26"}
  • {"month"=>"3", "year"=>"2017", "pdf_views"=>"3", "xml_views"=>"0", "html_views"=>"43"}
  • {"month"=>"4", "year"=>"2017", "pdf_views"=>"5", "xml_views"=>"0", "html_views"=>"22"}
  • {"month"=>"5", "year"=>"2017", "pdf_views"=>"18", "xml_views"=>"0", "html_views"=>"34"}
  • {"month"=>"6", "year"=>"2017", "pdf_views"=>"1", "xml_views"=>"0", "html_views"=>"29"}
  • {"month"=>"7", "year"=>"2017", "pdf_views"=>"9", "xml_views"=>"3", "html_views"=>"24"}
  • {"month"=>"8", "year"=>"2017", "pdf_views"=>"6", "xml_views"=>"1", "html_views"=>"23"}
  • {"month"=>"9", "year"=>"2017", "pdf_views"=>"2", "xml_views"=>"1", "html_views"=>"11"}
  • {"month"=>"10", "year"=>"2017", "pdf_views"=>"3", "xml_views"=>"1", "html_views"=>"15"}
  • {"month"=>"11", "year"=>"2017", "pdf_views"=>"6", "xml_views"=>"0", "html_views"=>"57"}
  • {"month"=>"12", "year"=>"2017", "pdf_views"=>"2", "xml_views"=>"1", "html_views"=>"16"}
  • {"month"=>"1", "year"=>"2018", "pdf_views"=>"6", "xml_views"=>"1", "html_views"=>"15"}
  • {"month"=>"2", "year"=>"2018", "pdf_views"=>"4", "xml_views"=>"0", "html_views"=>"12"}
  • {"month"=>"3", "year"=>"2018", "pdf_views"=>"3", "xml_views"=>"0", "html_views"=>"8"}
  • {"month"=>"4", "year"=>"2018", "pdf_views"=>"4", "xml_views"=>"0", "html_views"=>"4"}
  • {"month"=>"5", "year"=>"2018", "pdf_views"=>"2", "xml_views"=>"0", "html_views"=>"4"}
  • {"month"=>"6", "year"=>"2018", "pdf_views"=>"4", "xml_views"=>"1", "html_views"=>"5"}
  • {"month"=>"7", "year"=>"2018", "pdf_views"=>"6", "xml_views"=>"4", "html_views"=>"23"}
  • {"month"=>"8", "year"=>"2018", "pdf_views"=>"3", "xml_views"=>"1", "html_views"=>"6"}
  • {"month"=>"9", "year"=>"2018", "pdf_views"=>"6", "xml_views"=>"0", "html_views"=>"14"}
  • {"month"=>"10", "year"=>"2018", "pdf_views"=>"3", "xml_views"=>"1", "html_views"=>"7"}
  • {"month"=>"11", "year"=>"2018", "pdf_views"=>"5", "xml_views"=>"0", "html_views"=>"3"}
  • {"month"=>"12", "year"=>"2018", "pdf_views"=>"6", "xml_views"=>"0", "html_views"=>"7"}
  • {"month"=>"1", "year"=>"2019", "pdf_views"=>"3", "xml_views"=>"0", "html_views"=>"4"}
  • {"month"=>"2", "year"=>"2019", "pdf_views"=>"3", "xml_views"=>"0", "html_views"=>"2"}
  • {"month"=>"3", "year"=>"2019", "pdf_views"=>"4", "xml_views"=>"0", "html_views"=>"9"}
  • {"month"=>"4", "year"=>"2019", "pdf_views"=>"4", "xml_views"=>"0", "html_views"=>"5"}
  • {"month"=>"5", "year"=>"2019", "pdf_views"=>"5", "xml_views"=>"0", "html_views"=>"11"}
  • {"month"=>"6", "year"=>"2019", "pdf_views"=>"3", "xml_views"=>"0", "html_views"=>"4"}
  • {"month"=>"7", "year"=>"2019", "pdf_views"=>"5", "xml_views"=>"0", "html_views"=>"7"}
  • {"month"=>"8", "year"=>"2019", "pdf_views"=>"4", "xml_views"=>"0", "html_views"=>"1"}

Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/593623"], "description"=>"<p>A. Stability of type I collagen mRNAs in WF-A treated HLF. HLF treated with DMSO (upper left panel) or 1.5 µM WF-A (upper right panel) for 24 h, transcription was blocked with Act D and collagen α1(I) and α2(I) mRNAs were measured by RT-PCR at the indicated times after the transcription block. Actin mRNA was measured as a loading control. Bottom panels: decay rates of collagen α1(I) and α2(I) mRNAs were estimated in DMSO and WF-A treated cells in three independent experiments. Expression of collagen mRNAs was normalized to the expression of actin mRNA and plotted as function of time after the transcription block. The levels at time 0 were arbitrarily set as 1. Error bars represent ±1SEM. B. Stability of collagen mRNAs in WF-A treated SCL fibroblasts. The experiments as in A, except SCL fibroblasts were used. * represents p<0.05.</p>", "links"=>[], "tags"=>["destabilizes", "collagen", "mrnas"], "article_id"=>264119, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042989.g003", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_WF_A_destabilizes_collagen_945_1_I_and_945_2_I_mRNAs_in_human_fibroblasts_/264119", "title"=>"WF-A destabilizes collagen α1(I) and α2(I) mRNAs in human fibroblasts.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-08-10 01:08:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/593962"], "description"=>"<p>A. WF-A inhibits TGF-β1 induced Smad3 phosphorylation. HLF were treated with DMSO (left panel) or WF-A (1.5 µM) (right panel) and stimulated by TGF-β1. Phospho-Smad3 and total Smad3 were measured by Western blot after indicated times. Loading control: actin. B. WF-A inhibits constitutive Smad3 phosphorylation in a dose dependent manner. Similar experiment as in A, HLF were treated with WF-A (0–1.5 µM). Loading control: actin. C. WF-A does not affect the amount of active TGF-β1 produced by HLF. HLF were treated with WF-A (0–1.5 µM) for 24 h. Conditioned medium from HLF was added to Ming lung epithelial cells (MLEC) stably transfected with TGF-β1 luciferase responsive promoter (PIA-Luc). Luciferase activity was measured in MLEC lysates. Gray bar: unconditioned medium, black bars: HLF conditioned medium, white bars: MLEC treated with recombinant TGF-β1 (0.05–0.8 ng/ml). Each bar represents three independent experiments analyzed in duplicate. Error bars: ±1SEM. D. Effect WF-A on expression of TGF-β1 relevant genes. mRNA of collagen III (COL α1(III)), fibronectin (FIB), Smad2, TGF-β1, TGF-β receptor 1 (TGFβR1) and TGF-β receptor 2 (TGFβR2) from control cells and HLF treated with WF-A (0.75–1.5 µM) were analyzed by RT-PCR (left panel). Protein COLα1(III), FIB, TGFβR1 and phospho-TGFβR1 were analyzed by Western blot (right panel). Loading control: actin. E. Synergistic effect of p38 MAPK inhibitor (SB203580) and WF-A. HLF were treated with SB203580 (10 µM, lane 2), WF-A (1.5 µM, lane 3) or both (lane 4), DMSO (lane 1). COLα1 and COLα2 mRNAs were analyzed after 24 h by RT-PCR. Loading control: actin mRNA. Bottom panel: expression of collagen α1(I) mRNA (open bars) and α2(I) mRNA (black bars) were normalized to actin mRNA and plotted from three independent experiments. The error bars: ±1SEM. F. Effect of SB203580 and WF-A in SCL. Similar experiment as in E. * represents p<0.05 and # represents p<0.01.</p>", "links"=>[], "tags"=>["inhibits"], "article_id"=>264455, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042989.g006", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_WF_A_inhibits_TGF_946_1_signaling_/264455", "title"=>"WF-A inhibits TGF-β1 signaling.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-08-10 01:14:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/594195"], "description"=>"<p>A. Mice were injected with isoproterenol (ISO), isoproterenol + WF-A (ISO+WF-A), WF-A alone or vehicle (CON). Heart weight (HW) to body weight (BW) ratio (HW:BW) is shown as the mean ±1 SEM. B. WF-A protects against necrosis of heart tissue. H&E staining of a representative section of a heart from the control group (CONT), isoproterenol group (ISO), isoproterenol + WF-A group (ISO + WF-A) and WF-A group (WF-A). Scale bar: 200 µm. C. WF-A decreases isoproterenol induced myocardial fibrosis in mice. Masson’s trichrome staining of the base of the ventricle sections of hearts of control mice and mice treated with ISO and with ISO+WF-A. Two representative sections of each heart are shown with collagen staining in blue. Scale bar: 2 mm. D. Morphometric analysis of the percentage of fibrotic area of the hearts. Fibrosis areas within the sections (blue) were quantified using Image J software. The percentage of fibrosis was estimated from the ventricular and apex sections of each heart. Data represent ±1 SEM of six mice per group. * represents p<0.05 and # represents p<0.01.</p>", "links"=>[], "tags"=>["inhibits", "isoproterenol", "induced", "myocardial", "fibrosis"], "article_id"=>264681, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042989.g008", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_WF_A_inhibits_isoproterenol_induced_myocardial_fibrosis_in_mice_/264681", "title"=>"WF-A inhibits isoproterenol induced myocardial fibrosis in mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-08-10 01:18:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/594442"], "description"=>"<p>F, forward primer; R, reverse primer; h, human; m, mouse; r, rat; and SMA, smooth muscle actin.</p>", "links"=>[], "tags"=>["rt-pcr"], "article_id"=>264937, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042989.t001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Primers_used_for_RT_PCR_and_QRT_PCR_/264937", "title"=>"Primers used for RT-PCR and QRT-PCR.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-08-10 01:22:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/594067"], "description"=>"<p>HSC were isolated from rat livers and cultured for 2 days, when the cells were treated with DMSO (lane 1) or WF-A (0.25 µM, lane 2). At day 6, total RNA was analyzed for collagen α1(I) mRNA (COLα1), collagen α2(I) mRNA (COLα2) and α-smooth muscle actin mRNA (α-SMA) by RT-PCR. Loading controls: actin mRNA and GAPDH mRNA.</p>", "links"=>[], "tags"=>["inhibits", "activation"], "article_id"=>264556, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042989.g007", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_WF_A_inhibits_culture_activation_of_primary_rat_HSC_/264556", "title"=>"WF-A inhibits culture activation of primary rat HSC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-08-10 01:15:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/593397"], "description"=>"<p>A. Disruption of vimentin filaments in human fibroblasts treated with WF-A. Primary human lung fibroblasts (HLF, upper panels) and scleroderma fibroblasts (SCL, lower panels) were treated for 2 h with DMSO or with 1.0 µM of WF-A and immunostained with anti-vimentin antibody. Nuclear staining was by DAPI (blue). Bars, 1 µm. B. Collapse of vimentin filaments in rodent fibroblasts treated with WF-A. Experiment as in A, except primary rat cardiac fibroblasts (RCF, upper panels) and mouse embryonic fibroblasts (MEF, lower panels) were treated with DMSO or 1.0 µM of WF-A. Bars, 1 µm. C. Degradation of soluble vimentin by WF-A. Soluble vimentin was extracted with high salt buffer from HLF (left) and MEF (right) after treatment with the indicated concentrations of WF-A for 2 h. Vimentin (VIM) was analyzed by Western blot. Loading control: actin.</p>", "links"=>[], "tags"=>["wf-a", "vimentin", "filaments"], "article_id"=>263885, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042989.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_WF_A_on_integrity_of_vimentin_filaments_in_fibroblasts_/263885", "title"=>"Effect of WF-A on integrity of vimentin filaments in fibroblasts.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-08-10 01:04:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/593747"], "description"=>"<p>A. Expression of collagen polypeptides in vimentin knock-out fibroblasts treated with WF-A. Wild type mouse embryonic fibroblasts (VIM+/+ MEFs, upper right panel) and vimentin knock-out (VIM−/− MEFs, upper left panel) were treated with WF-A and collagen α1(I) and α2(I) polypeptides analyzed by Western blotting. Loading control: actin. Bottom panel: the expression of collagen α1(I) and α2(I) polypeptides from VIM+/+ MEFs (open bars) and VIM−/− MEFs (black bars) was normalized to actin expression andresults from three independent experiments were plotted. The error bars represent ±1SEM. B. Effect of WF-A on expression of collagen mRNAs in VIM+/+ and VIM−/− MEFs. Experiment as in A, except mRNAs were analyzed by RT-PCR. C. WF-A destabilizes collagen mRNAs in VIM+/+ MEFs. VIM+/+ MEFs were treated with DMSO (left panel) or 1.0 µM WF-A (right panel) for 24 h. Transcription was then blocked by Act-D and collagen α1(I) and α2(I) mRNAs were measured at the indicated times after the block by RT-PCR. Actin mRNA was measured as a loading control. Bottom panel: VIM+/+ MEFs treated with WF-A (full lines) or DMSO (broken lines) and the levels of collagen α1(I) mRNA (circles) and α2(I) mRNA (squares) were normalized to actin mRNA and plotted for the indicated time points after the transcription block. The level at time point 0 was arbitrarily set as 1. The error bars represent ±1 SEM, estimated from three independent experiments. D. WF-A has no effect on stability of collagen mRNAs in VIM−/− MEFs. Experiment as in C, except in VIM−/− MEFs were used. * represents p<0.05.</p>", "links"=>[], "tags"=>["wf-a", "collagen", "vimentin", "knock-out"], "article_id"=>264238, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042989.g004", "stats"=>{"downloads"=>1, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_WF_A_on_collagen_expression_in_vimentin_knock_out_fibroblasts_/264238", "title"=>"Effects of WF-A on collagen expression in vimentin knock-out fibroblasts.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-08-10 01:10:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/593845"], "description"=>"<p>A. WF-A inhibits TGF-β1 induced increase in collagen polypeptides. HLFs were treated with DMSO (lanes 1 and 2) or 1.5 µM WF-A (lane 3) for 1 h and then treated with TGF-β1 (lanes 2 and 3) for 24 h. Expression of collagen α1(I) (COLα1) and collagen α2(I) (COLα2) polypeptides in cell lysates was measured by Western blot. Loading control: actin. Bottom panel: levels of collagen α1(I) and α2(I) polypeptides were normalized to that of actin and plotted from three independent experiments. The error bars represent ±1 SEM. B. WF-A inhibits induction of collagen mRNAs by TGF-β1. Experiment as in A, except collagen mRNAs were analyzed by RT-PCR. Loading control: actin. Bottom panel: levels of collagen α1(I) and α2(I) mRNAs were normalized to that of actin mRNA and plotted from three independent experiments. The error bars represent ±1 SEM. C. WF-A inhibits transcriptional activation of collagen α2(I) promoter. Luciferase reporter gene containing wild type collagen α2(I) promoter (open bars) or collagen α2(I) promoter with the mutated TGF-β1-responsive element (black bars) were transfected into HLF. After treatment with WF-A and TGF-β1, luciferase activity was normalized to the co-transfected internal control gene β-GAL. The plot is from three independent transfections performed in duplicate. Error bars represent ±1 SEM. D. WF-A inhibits transcriptional activation of a SMAD responsive promoter. Experiment as in C, except reporter gene containing (CAGA)<sub>12</sub>MLP promoter was used and WF-A was used at concentrations 0.75 µM (gray bars) and 1.5 µM (black bars). * represents p<0.05 and # represents p<0.01.</p>", "links"=>[], "tags"=>["inhibits", "induced"], "article_id"=>264336, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042989.g005", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_WF_A_inhibits_TGF_946_1_induced_expression_of_type_I_collagen_/264336", "title"=>"WF-A inhibits TGF-β1 induced expression of type I collagen.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-08-10 01:12:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/593523"], "description"=>"<p>A. WF-A reduces collagen type I protein in human fibroblasts. HLF (upper left panel) and SCL fibroblasts (upper right panel) were treated WF-A and cellular levels of collagen α1(I) (COLα1) and α2(I) (COLα1) polypeptides were analyzed by Western blotting. Loading control: actin. Bottom panels; the expression of collagen polypeptides were normalized to actin and plotted from three independent experiments. Open bars: HLF, black bars: SCL. Low concentrations of WF-A: (0.75 µM for HLF) and (0.5 µM for SCL). High concentrations of WF-A: (1.5 µM for HLF) and (1.0 µM for SCL). The error bars: ±1 SEM are shown. B. WF-A decreases the steady-state levels of collagen α1 and α2 mRNAs in human fibroblasts. Experiment as in A, mRNAs were analyzed by RT-PCR (upper panels). Middle panel is quantification of RT-PCR from three independent experiments, and the bottom panel show quantification by real time (qRT-PCR). C. WF-A reduces collagen protein in primary rat hepatic stellate cells (HSC) and primary rat cardiac fibroblasts (RCF). HSC and RCF were isolated from rat liver and heart, respectively, and treated with the indicated amounts of WF-A. Experiment as in A. Loading control: actin. Bottom panels: levels of collagen α1(I) (COLα1) and collagen α2(I) (COLα2) polypeptides were normalized to actin and plotted from three independent experiments. Open bars: HSC, black bars: RCF. Low concentrations of WF-A: (0.75 µM for HSC) and (0.5 µM for RCF). High concentrations: (1.5 µM for HSC) and (1.0 µM for RCF). The error bars: ±1 SEM are shown. D. WF-A decreases the steady-state levels of collagen mRNAs in HSC and RCF. Experiment as in C, mRNAs were analyzed by RT-PCR (upper panels, with quantification from three experiments shown in middle panel) and qRT-PCR (bottom panel), * represents p<0.05.</p>", "links"=>[], "tags"=>["collagen", "polypeptides", "mrnas", "rodent"], "article_id"=>264017, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042989.g002", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_WF_A_reduces_expression_of_type_I_collagen_945_1_and_945_2_polypeptides_and_their_mRNAs_in_human_and_rodent_fibroblasts_/264017", "title"=>"WF-A reduces expression of type I collagen α1 and α2 polypeptides and their mRNAs in human and rodent fibroblasts.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-08-10 01:06:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/594365"], "description"=>"<p>A. WF-A decreased expression of collagen α1(I) and α2(I) mRNAs in experimental cardiac fibrosis. Total RNA was extracted from the heart tissue of the study groups and analyzed by RT-PCR for expression of collagen α1(I) mRNA (COLα1), collagen α2(I) mRNA (COLα2) and α-smooth muscle actin mRNA (α-SMA). Loading control: actin mRNA. The numbers indicate individual animals in each experimental group. Bottom panel: Expression of collagen α1(I) mRNA (open bars) and collagen α2(I) mRNA (black bars) was normalized to actin mRNA and plotted for each group. Error bars represent ±1SEM. B. WF-A inhibits expression of collagen α1(III) and α-smooth muscle actin mRNAs in isoproterenol induced cardiac fibrosis. Experiment as in A, except collagen α1(III) mRNA (COLIII) and α-smooth muscle actin mRNAs (α-SMA) mRNA were analyzed. C. WF-A reduces collagen α1(I) and α1(III) polypeptides and α-smooth muscle actin protein. Total protein extracted from the hearts of the study groups was analyzed for collagen α1(I) polypeptide, collagen α1(III) polypeptide and α-SMA protein by Western blot. Loading control: actin. The numbers represent individual animals in the experimental groups. Bottom panel: protein expression was normalized to actin expression and plotted for the groups. Collagen α1(I) polypeptide; open bars, collagen α1(III) polypeptide; black bars and α-SMA protein; gray bars. Error bars: ±1SEM. * represents p<0.05 and # represents p<0.01.</p>", "links"=>[], "tags"=>["inhibits", "iii", "collagens", "isoproterenol", "induced", "cardiac"], "article_id"=>264853, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042989.g009", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_WF_A_inhibits_expression_of_945_SMA_type_I_and_type_III_collagens_in_isoproterenol_induced_cardiac_fibrosis_/264853", "title"=>"WF-A inhibits expression of α-SMA, type I, and type III collagens in isoproterenol induced cardiac fibrosis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-08-10 01:20:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/310836", "https://ndownloader.figshare.com/files/310884", "https://ndownloader.figshare.com/files/310899"], "description"=>"<div><p>Type I collagen is the most abundant protein in the human body. Its excessive synthesis results in fibrosis of various organs. Fibrosis is a major medical problem without an existing cure. Excessive synthesis of type I collagen in fibrosis is primarily due to stabilization of collagen mRNAs. We recently reported that intermediate filaments composed of vimentin regulate collagen synthesis by stabilizing collagen mRNAs. Vimentin is a primary target of Withaferin-A (WF-A). Therefore, we hypothesized that WF-A may reduce type I collagen production by disrupting vimentin filaments and decreasing the stability of collagen mRNAs. This study is to determine if WF-A exhibits anti-fibrotic properties <em>in vitro</em> and <em>in vivo</em> and to elucidate the molecular mechanisms of its action. In lung, skin and heart fibroblasts WF-A disrupted vimentin filaments at concentrations of 0.5–1.5 µM and reduced 3 fold the half-lives of collagen α1(I) and α2(I) mRNAs and protein expression. In addition, WF-A inhibited TGF-β1 induced phosphorylation of TGF-β1 receptor I, Smad3 phosphorylation and transcription of collagen genes. WF-A also inhibited <em>in vitro</em> activation of primary hepatic stellate cells and decreased their type I collagen expression. In mice, administration of 4 mg/kg WF-A daily for 2 weeks reduced isoproterenol-induced myocardial fibrosis by 50%. Our findings provide strong evidence that Withaferin-A could act as an anti-fibrotic compound against fibroproliferative diseases, including, but not limited to, cardiac interstitial fibrosis.</p> </div>", "links"=>[], "tags"=>["withaferin-a", "reduces", "collagen", "vitro", "inhibits", "myocardial", "fibrosis", "vivo"], "article_id"=>121321, "categories"=>["Molecular Biology", "Biochemistry", "Cell Biology"], "users"=>["Azariyas A. Challa", "Milica Vukmirovic", "John Blackmon", "Branko Stefanovic"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0042989.s001", "https://dx.doi.org/10.1371/journal.pone.0042989.s002", "https://dx.doi.org/10.1371/journal.pone.0042989.s003"], "stats"=>{"downloads"=>2, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Withaferin_A_Reduces_Type_I_Collagen_Expression_In_Vitro_and_Inhibits_Development_of_Myocardial_Fibrosis_In_Vivo/121321", "title"=>"Withaferin-A Reduces Type I Collagen Expression In Vitro and Inhibits Development of Myocardial Fibrosis In Vivo", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-08-10 00:22:01"}

PMC Usage Stats | Further Information

  • {"unique-ip"=>"27", "full-text"=>"25", "pdf"=>"14", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"8"}
  • {"unique-ip"=>"53", "full-text"=>"50", "pdf"=>"41", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"20", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"9"}
  • {"unique-ip"=>"37", "full-text"=>"33", "pdf"=>"18", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"22", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"10"}
  • {"unique-ip"=>"26", "full-text"=>"28", "pdf"=>"11", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"12"}
  • {"unique-ip"=>"21", "full-text"=>"23", "pdf"=>"3", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"20", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"1"}
  • {"unique-ip"=>"18", "full-text"=>"23", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"13", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"3"}
  • {"unique-ip"=>"21", "full-text"=>"21", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"10", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"2"}
  • {"unique-ip"=>"16", "full-text"=>"14", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"12", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"4"}
  • {"unique-ip"=>"30", "full-text"=>"27", "pdf"=>"14", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"9", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"11"}
  • {"unique-ip"=>"18", "full-text"=>"16", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"5"}
  • {"unique-ip"=>"32", "full-text"=>"33", "pdf"=>"19", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"18", "supp-data"=>"6", "cited-by"=>"0", "year"=>"2013", "month"=>"6"}
  • {"unique-ip"=>"20", "full-text"=>"19", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2013", "month"=>"7"}
  • {"unique-ip"=>"23", "full-text"=>"23", "pdf"=>"12", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"8"}
  • {"unique-ip"=>"13", "full-text"=>"13", "pdf"=>"6", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"9"}
  • {"unique-ip"=>"14", "full-text"=>"13", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"10"}
  • {"unique-ip"=>"31", "full-text"=>"38", "pdf"=>"12", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"15", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"11"}
  • {"unique-ip"=>"9", "full-text"=>"11", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"12"}
  • {"unique-ip"=>"18", "full-text"=>"19", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"23", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"1"}
  • {"unique-ip"=>"18", "full-text"=>"15", "pdf"=>"12", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"2"}
  • {"unique-ip"=>"19", "full-text"=>"16", "pdf"=>"7", "abstract"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"3"}
  • {"unique-ip"=>"13", "full-text"=>"11", "pdf"=>"7", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"5"}
  • {"unique-ip"=>"15", "full-text"=>"20", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"6"}
  • {"unique-ip"=>"14", "full-text"=>"19", "pdf"=>"7", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"4"}
  • {"unique-ip"=>"10", "full-text"=>"14", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"17", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"4"}
  • {"unique-ip"=>"15", "full-text"=>"16", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"10", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2015", "month"=>"5"}
  • {"unique-ip"=>"17", "full-text"=>"17", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"10", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"6"}
  • {"unique-ip"=>"13", "full-text"=>"16", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2015", "month"=>"7"}
  • {"unique-ip"=>"15", "full-text"=>"17", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"3"}
  • {"unique-ip"=>"15", "full-text"=>"17", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"2"}
  • {"unique-ip"=>"11", "full-text"=>"14", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"8"}
  • {"unique-ip"=>"14", "full-text"=>"11", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"9"}
  • {"unique-ip"=>"12", "full-text"=>"13", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"10"}
  • {"unique-ip"=>"19", "full-text"=>"19", "pdf"=>"14", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"9", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"7"}
  • {"unique-ip"=>"11", "full-text"=>"6", "pdf"=>"5", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"8"}
  • {"unique-ip"=>"24", "full-text"=>"23", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"15", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"9"}
  • {"unique-ip"=>"13", "full-text"=>"10", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"10"}
  • {"unique-ip"=>"7", "full-text"=>"8", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"2"}
  • {"unique-ip"=>"12", "full-text"=>"9", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"11"}
  • {"unique-ip"=>"7", "full-text"=>"8", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"12"}
  • {"unique-ip"=>"7", "full-text"=>"14", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2015", "month"=>"1"}
  • {"unique-ip"=>"16", "full-text"=>"13", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"14", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2015", "month"=>"11"}
  • {"unique-ip"=>"9", "full-text"=>"12", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"12"}
  • {"unique-ip"=>"14", "full-text"=>"14", "pdf"=>"10", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"1"}
  • {"unique-ip"=>"12", "full-text"=>"15", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"3"}
  • {"unique-ip"=>"6", "full-text"=>"4", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"4"}
  • {"unique-ip"=>"8", "full-text"=>"7", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2016", "month"=>"5"}
  • {"unique-ip"=>"10", "full-text"=>"10", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2016", "month"=>"6"}
  • {"unique-ip"=>"11", "full-text"=>"14", "pdf"=>"11", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"7"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"8"}
  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"9"}
  • {"unique-ip"=>"16", "full-text"=>"17", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"10"}
  • {"unique-ip"=>"7", "full-text"=>"8", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"11"}
  • {"unique-ip"=>"7", "full-text"=>"7", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"12"}
  • {"unique-ip"=>"9", "full-text"=>"9", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2017", "month"=>"1"}
  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"2"}
  • {"unique-ip"=>"8", "full-text"=>"10", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"3"}
  • {"unique-ip"=>"7", "full-text"=>"7", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"12", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"4"}
  • {"unique-ip"=>"9", "full-text"=>"9", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"5"}
  • {"unique-ip"=>"8", "full-text"=>"8", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"6"}
  • {"unique-ip"=>"11", "full-text"=>"14", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"7"}
  • {"unique-ip"=>"7", "full-text"=>"8", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"8"}
  • {"unique-ip"=>"9", "full-text"=>"7", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"9"}
  • {"unique-ip"=>"10", "full-text"=>"10", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"10"}
  • {"unique-ip"=>"10", "full-text"=>"9", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"11"}
  • {"unique-ip"=>"8", "full-text"=>"10", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"12"}
  • {"unique-ip"=>"12", "full-text"=>"16", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"17", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"1"}
  • {"unique-ip"=>"1", "full-text"=>"1", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"2"}
  • {"unique-ip"=>"9", "full-text"=>"7", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"20", "supp-data"=>"0", "cited-by"=>"2", "year"=>"2018", "month"=>"3"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"9", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"1"}
  • {"unique-ip"=>"9", "full-text"=>"9", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"4"}
  • {"unique-ip"=>"5", "full-text"=>"6", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"5"}
  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"6"}
  • {"unique-ip"=>"13", "full-text"=>"15", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2018", "month"=>"7"}
  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"10"}
  • {"unique-ip"=>"5", "full-text"=>"6", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
  • {"unique-ip"=>"13", "full-text"=>"14", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"8"}
  • {"unique-ip"=>"14", "full-text"=>"15", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2018", "month"=>"9"}
  • {"unique-ip"=>"11", "full-text"=>"11", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
  • {"unique-ip"=>"79", "full-text"=>"79", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"17", "supp-data"=>"3", "cited-by"=>"1", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"7", "full-text"=>"7", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"5", "full-text"=>"7", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"8", "full-text"=>"9", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}

Relative Metric

{"start_date"=>"2012-01-01T00:00:00Z", "end_date"=>"2012-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences/Developmental biology", "average_usage"=>[313, 562, 690, 796, 893, 986, 1079, 1173, 1257, 1349, 1429, 1506, 1586, 1661, 1730, 1803, 1876, 1945, 2017, 2091, 2163, 2225, 2294, 2362, 2430]}, {"subject_area"=>"/Medicine and health sciences", "average_usage"=>[312, 547, 668, 769, 861, 953, 1046, 1135, 1224, 1307, 1388, 1464, 1536, 1606, 1676, 1744, 1812, 1882, 1954, 2020, 2089, 2155, 2218, 2282, 2344]}, {"subject_area"=>"/Medicine and health sciences/Anatomy", "average_usage"=>[307, 529, 649, 751, 845, 936, 1025, 1113, 1197, 1284, 1360, 1432, 1501, 1570, 1632, 1697, 1765, 1834, 1904, 1972, 2041, 2107, 2170, 2231, 2292]}]}
Loading … Spinner
There are currently no alerts