Combined Treatment with Troglitazone and Lovastatin Inhibited Epidermal Growth Factor-Induced Migration through the Downregulation of Cysteine-Rich Protein 61 in Human Anaplastic Thyroid Cancer Cells
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{"title"=>"Combined treatment with troglitazone and lovastatin inhibited epidermal growth factor-induced migration through the downregulation of cysteine-rich protein 61 in human anaplastic thyroid cancer cells", "type"=>"journal", "authors"=>[{"first_name"=>"Li Han", "last_name"=>"Chin", "scopus_author_id"=>"56540870400"}, {"first_name"=>"Sung Po", "last_name"=>"Hsu", "scopus_author_id"=>"35082050700"}, {"first_name"=>"Wen Bin", "last_name"=>"Zhong", "scopus_author_id"=>"57141520200"}, {"first_name"=>"Yu Chih", "last_name"=>"Liang", "scopus_author_id"=>"55950217200"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84924301380", "doi"=>"10.1371/journal.pone.0118674", "sgr"=>"84924301380", "pmid"=>"25742642", "issn"=>"19326203", "pui"=>"602684969"}, "id"=>"c851e972-2514-38b8-8bec-591824351ee8", "abstract"=>"Our previous studies have demonstrated that epidermal growth factor (EGF) can induce cell migration through the induction of cysteine-rich protein 61 (Cyr61) in human anaplastic thyroid cancer (ATC) cells. The aim of the present study was to determine the inhibitory effects of combined treatment with the peroxisome proliferator-activated receptor-γ (PPARγ) ligand troglitazone and the cholesterol-lowering drug lovastatin at clinically achievable concentrations on ATC cell migration. Combined treatment with 5 μM troglitazone and 1 μM lovastatin exhibited no cytotoxicity but significantly inhibited EGF-induced migration, as determined using wound healing and Boyden chamber assays. Cotreatment with troglitazone and lovastatin altered the epithelial-to-mesenchymal-transition (EMT) -related marker gene expression of the cells; specifically, E-cadherin expression increased and vimentin expression decreased. In addition, cotreatment reduced the number of filopodia, which are believed to be involved in migration, and significantly inhibited EGF-induced Cyr61 mRNA and protein expression as well as Cyr61 secretion. Moreover, the phosphorylation levels of 2 crucial signal molecules for EGF-induced Cyr61 expression, the cAMP response element-binding protein (CREB) and extracellular signal-regulated kinase (ERK), were decreased in cells cotreated with troglitazone and lovastatin. Performing a transient transfection assay revealed that the combined treatment significantly suppressed Cyr61 promoter activity. These results suggest that combined treatment with low doses of troglitazone and lovastatin effectively inhibits ATC cell migration and may serve as a novel therapeutic strategy for metastatic ATC.", "link"=>"http://www.mendeley.com/research/combined-treatment-troglitazone-lovastatin-inhibited-epidermal-growth-factorinduced-migration-throug", "reader_count"=>7, "reader_count_by_academic_status"=>{"Student > Doctoral Student"=>1, "Researcher"=>1, "Student > Postgraduate"=>1, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Student > Doctoral Student"=>1, "Researcher"=>1, "Student > Postgraduate"=>1, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>1, "Medicine and Dentistry"=>1, "Agricultural and Biological Sciences"=>2, "Veterinary Science and Veterinary Medicine"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Unspecified"=>{"Unspecified"=>1}, "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"=>{"Netherlands"=>1}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1941740"], "description"=>"<p>EGF treatment increases Cyr61 expression through the ERK/CREB signal pathways and then promotes cell migration. Combined treatment with troglitazone and lovastatin downregulates CREB activity by inhibiting the ERK and other unknown signaling pathways, eventually reducing Cyr61 expression and cell migration in ATC cells. Trog, troglitazone; Lova, lovastatin.</p>", "links"=>[], "tags"=>["vimentin expression", "Epidermal growth factor", "egf", "cells cotreated", "anaplastic thyroid cancer", "Boyden chamber assays", "phosphorylation levels", "Human Anaplastic Thyroid Cancer Cells", "signal molecules", "Cyr 61 promoter activity", "emt", "metastatic ATC", "troglitazone", "wound healing", "creb", "marker gene expression", "ppar", "ATC cell migration", "transfection assay", "erk", "cell migration", "drug lovastatin", "Protein expression", "Cyr 61 secretion"], "article_id"=>1329984, "categories"=>["Biological Sciences"], "users"=>["Li-Han Chin", "Sung-Po Hsu", "Wen-Bin Zhong", "Yu-Chih Liang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0118674.g008", "stats"=>{"downloads"=>0, "page_views"=>23, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Proposed_model_of_the_effects_of_combined_treatment_with_troglitazone_and_lovastatin_on_the_inhibition_of_EGF_induced_cell_migration_in_ATC_cells_/1329984", "title"=>"Proposed model of the effects of combined treatment with troglitazone and lovastatin on the inhibition of EGF-induced cell migration in ATC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-05 11:03:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1941738"], "description"=>"<p><b>A–C.</b> SW1736 cells were treated with troglitazone (5 μM) and lovastatin (1 μM) for 4 h and subsequently treated wtih EGF (10 ng/mL) for (A) 15 min, (B) 30 min, or (C) 2 min. Total protein was collected to detect the phosphorylation levels of (A) ERK, (B) CREB, and (C) EGFR by performing western blot analysis. Trog, troglitazone; Lova, lovastatin; and T/L, combined treatment with troglitazone and lovastatin.</p>", "links"=>[], "tags"=>["vimentin expression", "Epidermal growth factor", "egf", "cells cotreated", "anaplastic thyroid cancer", "Boyden chamber assays", "phosphorylation levels", "Human Anaplastic Thyroid Cancer Cells", "signal molecules", "Cyr 61 promoter activity", "emt", "metastatic ATC", "troglitazone", "wound healing", "creb", "marker gene expression", "ppar", "ATC cell migration", "transfection assay", "erk", "cell migration", "drug lovastatin", "Protein expression", "Cyr 61 secretion"], "article_id"=>1329982, "categories"=>["Biological Sciences"], "users"=>["Li-Han Chin", "Sung-Po Hsu", "Wen-Bin Zhong", "Yu-Chih Liang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0118674.g006", "stats"=>{"downloads"=>7, "page_views"=>89, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_combined_treatment_with_troglitazone_and_lovastatin_on_EGF_induced_phosphorylation_of_ERK_CREB_and_EGFR_in_ATC_cells_/1329982", "title"=>"Effects of combined treatment with troglitazone and lovastatin on EGF-induced phosphorylation of ERK, CREB, and EGFR in ATC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-05 11:03:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1941735"], "description"=>"<p><b>A–B.</b> SW1736 cells were cotreated with troglitazone (5 μM) and lovastatin (1 μM) for 3.5 h and subsequently treated with EGF (10 ng/mL) for 1 h. The cells were fixed, and immunocytochemistry staining was performed using phalloidin (red) to detect actin polymerization and DAPI staining was performed to detect nuclei (blue). <b>A.</b> The average number of filopodia per cell was determined by counting 30 cells. <b>B.</b> The figure depicts representative images. The arrows indicate the membrane ruffles and filopodia, and the representative regular membrane (open arrow) and filopodia (closed arrow) are shown. Con, control group and T/L, combined treatment with troglitazone and lovastatin.</p>", "links"=>[], "tags"=>["vimentin expression", "Epidermal growth factor", "egf", "cells cotreated", "anaplastic thyroid cancer", "Boyden chamber assays", "phosphorylation levels", "Human Anaplastic Thyroid Cancer Cells", "signal molecules", "Cyr 61 promoter activity", "emt", "metastatic ATC", "troglitazone", "wound healing", "creb", "marker gene expression", "ppar", "ATC cell migration", "transfection assay", "erk", "cell migration", "drug lovastatin", "Protein expression", "Cyr 61 secretion"], "article_id"=>1329979, "categories"=>["Biological Sciences"], "users"=>["Li-Han Chin", "Sung-Po Hsu", "Wen-Bin Zhong", "Yu-Chih Liang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0118674.g003", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_combined_treatment_with_troglitazone_and_lovastatin_on_EGF_induced_stress_fiber_formation_in_ATC_cells_/1329979", "title"=>"Effects of combined treatment with troglitazone and lovastatin on EGF-induced stress-fiber formation in ATC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-05 11:03:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1941734"], "description"=>"<p><b>A.</b> SW1736 cells were treated with troglitazone (5 μM) and/or lovastatin (1 μM) for 7 h, and the migrated cells were detected using a wound healing assay. <b>B.</b> The SW1736 cells were preincubated with troglitazone (5 μM) and/or lovastatin (1 μM) for 0.5 h before EGF treatment. After 7 h, the migrated cells were detected using the wound healing assay. <b>C.</b> The SW1736 cells were cotreated with troglitazone (5 μM) and lovastatin (1 μM) for 4 h, and the cells were then seeded in the upper Transwell chamber and EGF (20 ng/mL) was added to the lower chamber as a chemoattractant for cell migration. After 18 h of incubation, the transmigrated cells were stained and counted. All data are presented as the mean ± SE of 3 independent experiments. *<i>p</i> < 0.05, compared with the control group; <sup>#</sup>, <i>p</i> < 0.05. Con, control group; Trog, troglitazone; Lova, lovastatin; and T/L, combined treatment with troglitazone and lovastatin.</p>", "links"=>[], "tags"=>["vimentin expression", "Epidermal growth factor", "egf", "cells cotreated", "anaplastic thyroid cancer", "Boyden chamber assays", "phosphorylation levels", "Human Anaplastic Thyroid Cancer Cells", "signal molecules", "Cyr 61 promoter activity", "emt", "metastatic ATC", "troglitazone", "wound healing", "creb", "marker gene expression", "ppar", "ATC cell migration", "transfection assay", "erk", "cell migration", "drug lovastatin", "Protein expression", "Cyr 61 secretion"], "article_id"=>1329978, "categories"=>["Biological Sciences"], "users"=>["Li-Han Chin", "Sung-Po Hsu", "Wen-Bin Zhong", "Yu-Chih Liang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0118674.g002", "stats"=>{"downloads"=>0, "page_views"=>45, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_combined_treatment_with_troglitazone_and_lovastatin_on_EGF_induced_cell_migration_in_ATC_cells_/1329978", "title"=>"Effects of combined treatment with troglitazone and lovastatin on EGF-induced cell migration in ATC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-05 11:03:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1941737"], "description"=>"<p><b>A–B</b>. SW1736 cells were cotreated with troglitazone (5 μM) and lovastatin (1 μM) for 4 h and subsequently treated with EGF (10 ng/mL) for the indicated duration. <b>A.</b> Total RNA was collected for detecting the mRNA levels of Cyr61 by using real-time RT-PCR, and <b>B.</b> the total cellular protein was collected to detect the protein levels of Cyr61 by performing western blot analysis. <b>C.</b> The SW1736 cells were cotreated with troglitazone (5 μM) and lovastatin (1 μM) for 0.5 h and subsequently treated with EGF (20 ng/mL) for an additional 1 h. The cultured medium was collected to determine the Cyr61 protein levels by using an ELISA kit. The data are presented as the mean ± SE of 3 independent experiments. *<i>p</i> < 0.05, compared with the control group. <sup>#</sup><i>p</i> < 0.05. T/L, combined treatment with troglitazone and lovastatin.</p>", "links"=>[], "tags"=>["vimentin expression", "Epidermal growth factor", "egf", "cells cotreated", "anaplastic thyroid cancer", "Boyden chamber assays", "phosphorylation levels", "Human Anaplastic Thyroid Cancer Cells", "signal molecules", "Cyr 61 promoter activity", "emt", "metastatic ATC", "troglitazone", "wound healing", "creb", "marker gene expression", "ppar", "ATC cell migration", "transfection assay", "erk", "cell migration", "drug lovastatin", "Protein expression", "Cyr 61 secretion"], "article_id"=>1329981, "categories"=>["Biological Sciences"], "users"=>["Li-Han Chin", "Sung-Po Hsu", "Wen-Bin Zhong", "Yu-Chih Liang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0118674.g005", "stats"=>{"downloads"=>0, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_combined_treatment_with_troglitazone_and_lovastatin_on_EGF_induced_Cyr61_expression_in_ATC_cells_/1329981", "title"=>"Effects of combined treatment with troglitazone and lovastatin on EGF-induced Cyr61 expression in ATC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-05 11:03:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1941736"], "description"=>"<p><b>A–C</b>. SW1736 cells were cotreated with troglitazone (5 μM) and lovastatin (1 μM) for 1 h and subsequently treated with EGF (10 ng/mL) for an additional 24 h. <b>A.</b> The cells were photographed, and representative pictures are shown. The (B) protein and (C) mRNA levels of E-cadherin and vimentin were determined by performing western blot analysis and real-time RT-PCR, respectively. All data are presented as the mean ± SE of 3 independent experiments. *, <i>p</i> < 0.05, compared with the control group; <sup>#</sup>, <i>p</i> < 0.05. T/L, combined treatment with troglitazone and lovastatin.</p>", "links"=>[], "tags"=>["vimentin expression", "Epidermal growth factor", "egf", "cells cotreated", "anaplastic thyroid cancer", "Boyden chamber assays", "phosphorylation levels", "Human Anaplastic Thyroid Cancer Cells", "signal molecules", "Cyr 61 promoter activity", "emt", "metastatic ATC", "troglitazone", "wound healing", "creb", "marker gene expression", "ppar", "ATC cell migration", "transfection assay", "erk", "cell migration", "drug lovastatin", "Protein expression", "Cyr 61 secretion"], "article_id"=>1329980, "categories"=>["Biological Sciences"], "users"=>["Li-Han Chin", "Sung-Po Hsu", "Wen-Bin Zhong", "Yu-Chih Liang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0118674.g004", "stats"=>{"downloads"=>2, "page_views"=>116, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_combined_treatment_with_troglitazone_and_lovastatin_on_EGF_induced_EMT_related_marker_protein_expression_in_ATC_cells_/1329980", "title"=>"Effects of combined treatment with troglitazone and lovastatin on EGF-induced EMT-related marker protein expression in ATC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-05 11:03:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1941733"], "description"=>"<p><b>A.</b> SW1736 human ATC cells were treated with the indicated concentrations of troglitazone and lovastatin for 24 h. <b>B.</b> WI-38 human fibroblast cells were cotreated with troglitazone (5 μM) and lovastatin (1 μM) for 24 h. Cell viability was determined using the crystal violet assay. The data are presented as the mean ± SEM of 3 independent experiments. *, <i>p</i> < 0.05, compared with the control group. Con, control group; Trog, troglitazone; Lova, lovastatin; and T/L, combined treatment with troglitazone and lovastatin.</p>", "links"=>[], "tags"=>["vimentin expression", "Epidermal growth factor", "egf", "cells cotreated", "anaplastic thyroid cancer", "Boyden chamber assays", "phosphorylation levels", "Human Anaplastic Thyroid Cancer Cells", "signal molecules", "Cyr 61 promoter activity", "emt", "metastatic ATC", "troglitazone", "wound healing", "creb", "marker gene expression", "ppar", "ATC cell migration", "transfection assay", "erk", "cell migration", "drug lovastatin", "Protein expression", "Cyr 61 secretion"], "article_id"=>1329977, "categories"=>["Biological Sciences"], "users"=>["Li-Han Chin", "Sung-Po Hsu", "Wen-Bin Zhong", "Yu-Chih Liang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0118674.g001", "stats"=>{"downloads"=>2, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_troglitazone_and_lovastatin_on_cell_viability_in_ATC_and_fibroblast_cells_/1329977", "title"=>"Effects of troglitazone and lovastatin on cell viability in ATC and fibroblast cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-05 11:03:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1941741"], "description"=>"<div><p>Our previous studies have demonstrated that epidermal growth factor (EGF) can induce cell migration through the induction of cysteine-rich protein 61 (Cyr61) in human anaplastic thyroid cancer (ATC) cells. The aim of the present study was to determine the inhibitory effects of combined treatment with the peroxisome proliferator-activated receptor-γ (PPARγ) ligand troglitazone and the cholesterol-lowering drug lovastatin at clinically achievable concentrations on ATC cell migration. Combined treatment with 5 μM troglitazone and 1 μM lovastatin exhibited no cytotoxicity but significantly inhibited EGF-induced migration, as determined using wound healing and Boyden chamber assays. Cotreatment with troglitazone and lovastatin altered the epithelial-to-mesenchymal-transition (EMT) -related marker gene expression of the cells; specifically, E-cadherin expression increased and vimentin expression decreased. In addition, cotreatment reduced the number of filopodia, which are believed to be involved in migration, and significantly inhibited EGF-induced Cyr61 mRNA and protein expression as well as Cyr61 secretion. Moreover, the phosphorylation levels of 2 crucial signal molecules for EGF-induced Cyr61 expression, the cAMP response element-binding protein (CREB) and extracellular signal-regulated kinase (ERK), were decreased in cells cotreated with troglitazone and lovastatin. Performing a transient transfection assay revealed that the combined treatment significantly suppressed Cyr61 promoter activity. These results suggest that combined treatment with low doses of troglitazone and lovastatin effectively inhibits ATC cell migration and may serve as a novel therapeutic strategy for metastatic ATC.</p></div>", "links"=>[], "tags"=>["vimentin expression", "Epidermal growth factor", "egf", "cells cotreated", "anaplastic thyroid cancer", "Boyden chamber assays", "phosphorylation levels", "Human Anaplastic Thyroid Cancer Cells", "signal molecules", "Cyr 61 promoter activity", "emt", "metastatic ATC", "troglitazone", "wound healing", "creb", "marker gene expression", "ppar", "ATC cell migration", "transfection assay", "erk", "cell migration", "drug lovastatin", "Protein expression", "Cyr 61 secretion"], "article_id"=>1329985, "categories"=>["Biological Sciences"], "users"=>["Li-Han Chin", "Sung-Po Hsu", "Wen-Bin Zhong", "Yu-Chih Liang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0118674", "stats"=>{"downloads"=>1, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Combined_Treatment_with_Troglitazone_and_Lovastatin_Inhibited_Epidermal_Growth_Factor_Induced_Migration_through_the_Downregulation_of_Cysteine_Rich_Protein_61_in_Human_Anaplastic_Thyroid_Cancer_Cells_/1329985", "title"=>"Combined Treatment with Troglitazone and Lovastatin Inhibited Epidermal Growth Factor-Induced Migration through the Downregulation of Cysteine-Rich Protein 61 in Human Anaplastic Thyroid Cancer Cells", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-05 11:03:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1941739"], "description"=>"<p>SW1736 cells were transfected with the Cyr61 reporter plasmid and phRL-TK plasmid as internal controls. The transfected SW1376 cells were then cotreated with troglitazone (5 μM) and lovastatin (1 μM) for 0.5 h and subsequently treated with EGF (10 ng/mL) for an additional 4 h. Total protein was collected to detect the Cyr61-promoter activity by using dual luciferase activity assay kits. The data are presented as the mean ± SE of 3 independent experiments. *, <i>p</i> < 0.05, compared with the control group; <sup>#</sup>, <i>p</i> < 0.05. T/L, combined treatment with troglitazone and lovastatin.</p>", "links"=>[], "tags"=>["vimentin expression", "Epidermal growth factor", "egf", "cells cotreated", "anaplastic thyroid cancer", "Boyden chamber assays", "phosphorylation levels", "Human Anaplastic Thyroid Cancer Cells", "signal molecules", "Cyr 61 promoter activity", "emt", "metastatic ATC", "troglitazone", "wound healing", "creb", "marker gene expression", "ppar", "ATC cell migration", "transfection assay", "erk", "cell migration", "drug lovastatin", "Protein expression", "Cyr 61 secretion"], "article_id"=>1329983, "categories"=>["Biological Sciences"], "users"=>["Li-Han Chin", "Sung-Po Hsu", "Wen-Bin Zhong", "Yu-Chih Liang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0118674.g007", "stats"=>{"downloads"=>1, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_combined_treatment_with_troglitazone_and_lovastatin_on_EGF_induced_Cyr61_gene_promoter_activity_in_ATC_cells_/1329983", "title"=>"Effects of combined treatment with troglitazone and lovastatin on EGF-induced Cyr61 gene promoter activity in ATC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-05 11:03:20"}

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  • {"unique-ip"=>"7", "full-text"=>"5", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}

Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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