Responses of Human Endothelial Cells to Pathogenic and Non-Pathogenic Leptospira Species
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Mendeley | Further Information

{"title"=>"Responses of human endothelial cells to pathogenic and non-pathogenic Leptospira species", "type"=>"journal", "authors"=>[{"first_name"=>"Denise G.", "last_name"=>"Martinez-Lopez", "scopus_author_id"=>"56549028300"}, {"first_name"=>"Mark", "last_name"=>"Fahey", "scopus_author_id"=>"57197338388"}, {"first_name"=>"Jenifer", "last_name"=>"Coburn", "scopus_author_id"=>"35744632800"}], "year"=>2010, "source"=>"PLoS Neglected Tropical Diseases", "identifiers"=>{"sgr"=>"78650714951", "doi"=>"10.1371/journal.pntd.0000918", "pui"=>"361011524", "pmid"=>"21179504", "scopus"=>"2-s2.0-78650714951", "issn"=>"1935-2735", "isbn"=>"1935-2727"}, "id"=>"7ecb9b39-3dc4-3842-bce9-43af3b2b077d", "abstract"=>"Leptospirosis is a widespread zoonotic infection that primarily affects residents of tropical regions, but causes infections in animals and humans in temperate regions as well. The agents of leptospirosis comprise several members of the genus Leptospira, which also includes non-pathogenic, saprophytic species. Leptospirosis can vary in severity from a mild, non-specific illness to severe disease that includes multi-organ failure and widespread endothelial damage and hemorrhage. To begin to investigate how pathogenic leptospires affect endothelial cells, we compared the responses of two endothelial cell lines to infection by pathogenic versus non-pathogenic leptospires. Microarray analyses suggested that pathogenic L. interrogans and non-pathogenic L. biflexa triggered changes in expression of genes whose products are involved in cellular architecture and interactions with the matrix, but that the changes were in opposite directions, with infection by L. biflexa primarily predicted to increase or maintain cell layer integrity, while L. interrogans lead primarily to changes predicted to disrupt cell layer integrity. Neither bacterial strain caused necrosis or apoptosis of the cells even after prolonged incubation. The pathogenic L. interrogans, however, did result in significant disruption of endothelial cell layers as assessed by microscopy and the ability of the bacteria to cross the cell layers. This disruption of endothelial layer integrity was abrogated by addition of the endothelial protective drug lisinopril at physiologically relevant concentrations. These results suggest that, through adhesion of L. interrogans to endothelial cells, the bacteria may disrupt endothelial barrier function, promoting dissemination of the bacteria and contributing to severe disease manifestations. In addition, supplementing antibiotic therapy with lisinopril or derivatives with endothelial protective activities may decrease the severity of leptospirosis.", "link"=>"http://www.mendeley.com/research/responses-human-endothelial-cells-pathogenic-nonpathogenic-leptospira-species-6", "reader_count"=>30, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>5, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>5, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>1, "Environmental Science"=>1, "Agricultural and Biological Sciences"=>17, "Medicine and Dentistry"=>5, "Veterinary Science and Veterinary Medicine"=>2, "Chemistry"=>1, "Immunology and Microbiology"=>2}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>5}, "Chemistry"=>{"Chemistry"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>17}, "Unspecified"=>{"Unspecified"=>1}, "Environmental Science"=>{"Environmental Science"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>2}}, "reader_count_by_country"=>{"United States"=>1, "Brazil"=>2}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/812806"], "description"=>"<p>Ea.hy926 endothelial cell layers were infected with <i>L. interrogans</i> sv. Copenhageni as described in <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000918#s2\" target=\"_blank\">Materials and Methods</a>, except that just prior to the addition of the bacteria the drugs lisinopril or telmisartan were added to the concentrations indicated. The micrographs were taken at the 3 hour time point. Lisinopril at 10 µM, 1 µM and 100 nM blocked endothelial disruption by <i>L. interrogans</i>; lisinopril at 10 nM or below did not.</p>", "links"=>[], "tags"=>["concentrations", "endothelial", "layers"], "article_id"=>483176, "categories"=>["Infectious Diseases"], "users"=>["Denise G. Martinez-Lopez", "Mark Fahey", "Jenifer Coburn"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000918.g006", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Lisinopril_concentrations_effective_in_protection_of_endothelial_cell_layers_from_damage_due_to_L_interrogans_/483176", "title"=>"Lisinopril concentrations effective in protection of endothelial cell layers from damage due to <i>L. interrogans</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 21:51:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/812923"], "description"=>"1<p>KEGG Pathways: <a href=\"http://www.genome.jp/kegg/pathway.html\" target=\"_blank\">http://www.genome.jp/kegg/pathway.html</a>. The Table is arranged in order of descending p value for <i>L. interrogans</i>. The comparisons were made between cells infected with the indicated bacterial strain vs. uninfected controls at 1 hr. post infection. Similar trends were observed at the 3 hr. time point. Pathway abbreviations are provided for reference to <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000918#pntd-0000918-t002\" target=\"_blank\">Table 2</a>. The same pathways also showed significant changes in expression, in similar patterns, in a second endothelial cell line, HMEC (data not shown).</p>2<p>P values considered to be statistically significant were <0.05 by the Hypergeometric test. <i>(</i><a href=\"http://bioinfo.vanderbilt.edu/webgestalt\" target=\"_blank\">http://bioinfo.vanderbilt.edu/webgestalt</a><i>)</i>.</p>", "links"=>[], "tags"=>["genes", "changes", "endothelial", "cells", "infected", "serovar", "patoc"], "article_id"=>483284, "categories"=>["Infectious Diseases"], "users"=>["Denise G. Martinez-Lopez", "Mark Fahey", "Jenifer Coburn"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000918.t001", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Clusters_of_3_genes_with_statistically_significant_changes_in_expression_common_to_Ea_hy926_endothelial_cells_infected_with_L_biflexa_serovar_Patoc_and_L_interrogans_serovar_Canicola_/483284", "title"=>"Clusters of ≥3 genes with statistically significant changes in expression common to Ea.hy926 endothelial cells infected with <i>L. biflexa</i> serovar Patoc and <i>L. interrogans</i> serovar Canicola.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 21:52:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/812297"], "description"=>"<p>Ea.hy926 endothelial cells were plated in tissue culture treated glass chamber slides and allowed to reach near confluence (assessed visually). The bacteria were added at MOI = 10 and incubated with the endothelial cells for 1 or 3 hours at 37°C, then were washed and fixed. The slides were stained with phalloidin-FITC, which illuminates F actin, plus anti-<i>Leptospira</i> antibodies followed by TRITC- conjugated secondary antibody. Retraction of the cell bodies in response to <i>L. interrogans</i> Canicola, but not <i>L. biflexa</i> Patoc, is evident, particularly at 3 hr infection. The brighter staining of rounded and retracted cells with FITC-phalloidin may be due to disorganization of cellular architecture without complete depolymerization of the actin, which in the increased depth and decreased area of the cytoplasm would appear more concentrated and therefore brighter. Changes in endothelial cell morphology were most evident, and at earlier time points, in cells with which the <i>L. interrogans</i> bacteria were associated. One higher magnification micrograph of <i>L. interrogans</i> Canicola infected cells is included because the bacteria are small when viewing fields of endothelial cells that provide information on integrity of the monolayer. Micrographs are representative of multiple (>12) independent experiments. <i>L. interrogans</i> Copenhageni caused essentially the same changes in endothelial cell morphology as <i>L. interrogans</i> Canicola (data not shown).</p>", "links"=>[], "tags"=>["causes", "disruptions", "endothelial"], "article_id"=>482662, "categories"=>["Infectious Diseases"], "users"=>["Denise G. Martinez-Lopez", "Mark Fahey", "Jenifer Coburn"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000918.g001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_L_interrogans_causes_disruptions_in_endothelial_cell_monolayers_/482662", "title"=>"<i>L. interrogans</i> causes disruptions in endothelial cell monolayers.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 21:48:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/812397"], "description"=>"<p>Ea.hy926 cells were infected with <i>L. interrogans</i> Canicola at MOI = 10 for 24 hr, then stained with the TUNEL-based ApoAlert DNA fragmentation kit (Clontech Laboratories, Inc.) according to the manufacturer's instructions. As a positive control, the cells were permeabilized, then treated with DNase1 at 1 µg/ml for 10 min at ambient temperature prior to staining. TUNEL staining (green nuclei) indicates DNA fragmentation consistent with apoptosis. Nuclei are also stained with propidium iodide (red), and the F actin cytoskeleton is stained with phalloidin (blue). The micrographs do not demonstrate any indication of apoptosis in <i>L. interrogans</i>-infected cells, and are representative of multiple experiments using multiple different tests for apoptosis, each performed at least three times.</p>", "links"=>[], "tags"=>["apoptosis", "endothelial"], "article_id"=>482772, "categories"=>["Infectious Diseases"], "users"=>["Denise G. Martinez-Lopez", "Mark Fahey", "Jenifer Coburn"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000918.g002", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_L_interrogans_infection_does_not_trigger_apoptosis_in_endothelial_cells_/482772", "title"=>"<i>L. interrogans</i> infection does not trigger apoptosis in endothelial cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 21:49:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/812477"], "description"=>"<p>Ea.hy926 and HMEC endothelial cells were placed in 3 µm transwell inserts (“upper chambers”) in 24 well dishes containing medium and allowed to reach complete confluence (assessed visually and with FITC-dextran 40,000). The bacteria were added at MOI = 50 (providing sufficient numbers for quantification) to wells with and without endothelial cells, and samples were taken for counting from both chambers of the transwell plates at the times indicated. The graphs show bacteria that have migrated from upper to lower chamber (i.e. transmigration) as the percent of the total in both the upper and lower chambers. Shown are the means ± standard deviations of 3 independent experiments. For <i>L. interrogans</i> sv. Copenhageni and Canicola, the rates of transmigration through membranes with and without cells were not statistically significant as assessed by repeated measures ANOVA followed by Bonferroni's multiple comparison test (P>0.05). For <i>L. biflexa</i>, the same comparison was significantly different, P<0.001. In addition, the <i>L. interrogans</i> strains were significantly different from <i>L. biflexa</i> in crossing the cell layers (P<0.05).</p>", "links"=>[], "tags"=>["transmigrates", "endothelial", "monolayers", "efficiently"], "article_id"=>482845, "categories"=>["Infectious Diseases"], "users"=>["Denise G. Martinez-Lopez", "Mark Fahey", "Jenifer Coburn"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000918.g003", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_L_interrogans_transmigrates_across_endothelial_cell_monolayers_more_efficiently_than_does_L_biflexa_/482845", "title"=>"<i>L. interrogans</i> transmigrates across endothelial cell monolayers more efficiently than does <i>L. biflexa</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 21:49:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/812957"], "description"=>"<p>The Table is arranged in order of descending differences between <i>L. interrogans</i> vs. <i>L. biflexa</i>-infected Ea.hy926 cells in comparison to the uninfected controls at I hr post-infection. For each gene, relevant KEGG pathway abbreviations are noted (see <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000918#pntd-0000918-t001\" target=\"_blank\">Table 1</a>).</p>", "links"=>[], "tags"=>["endothelial", "genes", "affected", "patoc", "canicola"], "article_id"=>483324, "categories"=>["Infectious Diseases"], "users"=>["Denise G. Martinez-Lopez", "Mark Fahey", "Jenifer Coburn"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000918.t002", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ea_hy926_endothelial_cell_genes_affected_differently_by_L_biflexa_Patoc_vs_L_interrogans_Canicola_infection_/483324", "title"=>"Ea.hy926 endothelial cell genes affected differently by <i>L. biflexa</i> Patoc vs. <i>L. interrogans</i> Canicola infection.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 21:52:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/812580"], "description"=>"<p>Ea.hy926 cell layers were treated with the proteoglycan synthesis inhibitor β-xyloside, or the control α-galactoside, as described in <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000918#pntd.0000918-Breiner1\" target=\"_blank\">[17]</a> prior to infection with <i>L. interrogans</i> sv. Copenhageni. After 3 hr, the cell layers were washed and fixed, then stained with phalloidin-FITC. Neither reagent significantly reduced disruption of the cell layers by <i>L. interrogans</i>, as alterations in cell morphology and significant gaps between cells were seen when <i>L. interrogans</i> was present, and trans-endothelial cell layer migration was not significantly affected (data not shown). Consistent with this result, chondroitin sulfates B and A, which do and do not inhibit <i>L. interrogans</i> attachment to mammalian cells, respectively <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000918#pntd.0000918-Breiner1\" target=\"_blank\">[17]</a>, also had no effect (not shown).</p>", "links"=>[], "tags"=>["proteoglycan", "binding", "disruption", "endothelial", "layers"], "article_id"=>482950, "categories"=>["Infectious Diseases"], "users"=>["Denise G. Martinez-Lopez", "Mark Fahey", "Jenifer Coburn"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000918.g004", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Interference_with_proteoglycan_binding_by_L_interrogans_does_not_prevent_disruption_of_endothelial_cell_layers_in_culture_/482950", "title"=>"Interference with proteoglycan binding by <i>L. interrogans</i> does not prevent disruption of endothelial cell layers in culture.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 21:50:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/812697"], "description"=>"<p>Panels A and B: Ea.hy926 endothelial cell layers were infected with <i>L. interrogans</i> sv. Copenhageni or <i>L. biflexa</i> sv. Patoc as described in <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000918#s2\" target=\"_blank\">Materials and Methods</a>, except that just prior to the addition of the bacteria the drugs lisinopril, telmisartan, dopamine, or furosemide were added to 1 µM. The micrographs shown in Panel A (representative of three experiments) were taken at the 6 hour time point; the graphs in Panel B show the transmigration of leptospires over the entire 72 hr. time course. Shown are the means and standard deviations of all data from three experiments. Statistical significance was determined using repeated measures ANOVA followed by Bonferroni's multiple comparison test. For wells with cells, <i>L. interrogans</i> vs. <i>L. biflexa</i>, p<0.001, <i>L. interrogans</i> with no additions vs. telmisartan p>0.05 (not significant), <i>L. interrogans</i> with no additions vs. lisinopril p<0.001, <i>L. interrogans</i> with telmisartan vs. lisinopril p<0.001. There were no significant differences in the absence of cells, and the drugs did not affect bacterial motility or attachment of <sup>35</sup>S-labeled leptospires to the cells (Panel C and data not shown).</p>", "links"=>[], "tags"=>["drugs", "endothelial", "caused"], "article_id"=>483071, "categories"=>["Infectious Diseases"], "users"=>["Denise G. Martinez-Lopez", "Mark Fahey", "Jenifer Coburn"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000918.g005", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_specific_drugs_that_protect_endothelial_barrier_function_on_damage_caused_by_L_interrogans_/483071", "title"=>"Effects of specific drugs that protect endothelial barrier function on damage caused by <i>L. interrogans</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 21:51:10"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"6", "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"}
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  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"5"}
  • {"unique-ip"=>"3", "full-text"=>"2", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"6"}
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  • {"unique-ip"=>"4", "full-text"=>"4", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"12"}
  • {"unique-ip"=>"6", "full-text"=>"5", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"1"}
  • {"unique-ip"=>"10", "full-text"=>"12", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"3"}
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  • {"unique-ip"=>"5", "full-text"=>"6", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"5"}
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  • {"unique-ip"=>"1", "full-text"=>"1", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
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  • {"unique-ip"=>"10", "full-text"=>"12", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"8", "full-text"=>"6", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
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  • {"unique-ip"=>"7", "full-text"=>"4", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"8", "full-text"=>"11", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"7", "full-text"=>"7", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}
  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"12"}
  • {"unique-ip"=>"10", "full-text"=>"10", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"2"}
  • {"unique-ip"=>"12", "full-text"=>"12", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"3"}
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Relative Metric

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