Humanized HLA-DR4 Mice Fed with the Protozoan Pathogen of Oysters Perkinsus Marinus (Dermo) Do Not Develop Noticeable Pathology but Elicit Systemic Immunity
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{"title"=>"Humanized HLA-DR4 mice fed with the protozoan pathogen of oysters Perkinsus marinus (Dermo) do not develop noticeable pathology but elicit systemic immunity", "type"=>"journal", "authors"=>[{"first_name"=>"Wathsala", "last_name"=>"Wijayalath", "scopus_author_id"=>"25634765800"}, {"first_name"=>"Sai", "last_name"=>"Majji", "scopus_author_id"=>"15754086500"}, {"first_name"=>"Yuliya", "last_name"=>"Kleschenko", "scopus_author_id"=>"55972044300"}, {"first_name"=>"Luis", "last_name"=>"Pow-Sang", "scopus_author_id"=>"56154605300"}, {"first_name"=>"Teodor D.", "last_name"=>"Brumeanu", "scopus_author_id"=>"6701775123"}, {"first_name"=>"Eileen Franke", "last_name"=>"Villasante", "scopus_author_id"=>"55649232700"}, {"first_name"=>"Gerardo R.", "last_name"=>"Vasta", "scopus_author_id"=>"35466274800"}, {"first_name"=>"José Antonio", "last_name"=>"Fernández-Robledo", "scopus_author_id"=>"23008530200"}, {"first_name"=>"Sofia", "last_name"=>"Casares", "scopus_author_id"=>"6701821265"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"373059532", "sgr"=>"84900318484", "doi"=>"10.1371/journal.pone.0087435", "scopus"=>"2-s2.0-84900318484", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"24498105"}, "id"=>"340d042f-3f5c-34d0-bd19-f4958c672047", "abstract"=>"Perkinsus marinus (Phylum Perkinsozoa) is a marine protozoan parasite responsible for \"Dermo\" disease in oysters, which has caused extensive damage to the shellfish industry and estuarine environment. The infection prevalence has been estimated in some areas to be as high as 100%, often causing death of infected oysters within 1-2 years post-infection. Human consumption of the parasites via infected oysters is thus likely to occur, but to our knowledge the effect of oral consumption of P. marinus has not been investigated in humans or other mammals. To address the question we used humanized mice expressing HLA-DR4 molecules and lacking expression of mouse MHC-class II molecules (DR4.EA(0)) in such a way that CD4 T cell responses are solely restricted by the human HLA-DR4 molecule. The DR4.EA(0) mice did not develop diarrhea or any detectable pathology in the gastrointestinal tract or lungs following single or repeated feedings with live P. marinus parasites. Furthermore, lymphocyte populations in the gut associated lymphoid tissue and spleen were unaltered in the parasite-fed mice ruling out local or systemic inflammation. Notably, naïve DR4.EA(0) mice had antibodies (IgM and IgG) reacting against P. marinus parasites whereas parasite specific T cell responses were undetectable. Feeding with P. marinus boosted the antibody responses and stimulated specific cellular (IFNγ) immunity to the oyster parasite. Our data indicate the ability of P. marinus parasites to induce systemic immunity in DR4.EA(0) mice without causing noticeable pathology, and support rationale grounds for using genetically engineered P. marinus as a new oral vaccine platform to induce systemic immunity against infectious agents.", "link"=>"http://www.mendeley.com/research/humanized-hladr4-mice-fed-protozoan-pathogen-oysters-perkinsus-marinus-dermo-not-develop-noticeable", "reader_count"=>19, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Student > Doctoral Student"=>3, "Researcher"=>1, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>1, "Student > Master"=>3, "Other"=>2, "Student > Bachelor"=>2, "Professor"=>3}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Student > Doctoral Student"=>3, "Researcher"=>1, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>1, "Student > Master"=>3, "Other"=>2, "Student > Bachelor"=>2, "Professor"=>3}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Environmental Science"=>1, "Agricultural and Biological Sciences"=>13, "Medicine and Dentistry"=>2, "Neuroscience"=>1, "Economics, Econometrics and Finance"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Neuroscience"=>{"Neuroscience"=>1}, "Economics, Econometrics and Finance"=>{"Economics, Econometrics and Finance"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>13}, "Environmental Science"=>{"Environmental Science"=>1}}, "reader_count_by_country"=>{"Italy"=>1, "South Africa"=>1}, "group_count"=>2}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1371698"], "description"=>"<p><b>Panel A, </b><i>P. marinus</i> parasites (7×10<sup>5</sup> to 9×10<sup>5</sup>) were cultured for 10 minutes in media adjusted to pH values ranging from 2.0–6.2 and cell viability was measured by trypan blue exclusion. Data represent mean ± SD of two independent experiments. P values are indicated over the plots. <b>Panel B,</b> DR4.EA<sup>0</sup> mice (n = 10) were fed with 10<sup>5</sup> live <i>P. marinus</i> and set in 4 clean cages (2–3 mice per cage). Feces were collected at 24 h and 48 h post-feeding. DNA (10 ng) extracted from fecal samples was amplified with a pair of primers targeting NTS domain located between 5S and SSU rRNA genes from <i>P. marinus</i> (307 bp amplicon). Upper panel shows absence of parasite DNA in feces at 24 h and 48 h post-feeding. Lower panel shows that increasing concentration of fecal DNA (10 ng/µl) did not result in detectable PCR signal. Mixture of fecal DNA and purified parasite DNA (2∶1) resulted in positive PCR signal (lower panel) ruling out that potential inhibitory components in fecal material could have led to false negative results. PC, positive control; NC, negative control; MW, DNA molecular markers; Mix, Mixture of fecal DNA and purified <i>P. marinus</i> genomic DNA.</p>", "links"=>[], "tags"=>["immunology", "Immune cells", "Antibody-producing cells", "b cells", "t cells", "Immune system", "Lymphoid organs", "immunity", "Adaptive immunity", "Humoral immunity", "immunizations", "Immune response", "immunoglobulins", "Major histocompatibility complex", "Marine biology", "Fisheries science", "Model organisms", "Animal models", "mouse", "gastric", "ph"], "article_id"=>922444, "categories"=>["Biological Sciences"], "users"=>["Wathsala Wijayalath", "Sai Majji", "Yuliya Kleschenko", "Luis Pow-Sang", "Teodor D. Brumeanu", "Eileen Franke Villasante", "Gerardo R. Vasta", "José-Antonio Fernández-Robledo", "Sofia Casares"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087435.g001", "stats"=>{"downloads"=>0, "page_views"=>23, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_P_marinus_is_sensitive_to_gastric_pH_and_does_not_shed_from_the_intestine_/922444", "title"=>"<i>P. marinus</i> is sensitive to gastric pH and does not shed from the intestine.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-31 03:38:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/1371718"], "description"=>"<div><p><i>Perkinsus marinus</i> (Phylum Perkinsozoa) is a marine protozoan parasite responsible for “Dermo” disease in oysters, which has caused extensive damage to the shellfish industry and estuarine environment. The infection prevalence has been estimated in some areas to be as high as 100%, often causing death of infected oysters within 1–2 years post-infection. Human consumption of the parasites <i>via</i> infected oysters is thus likely to occur, but to our knowledge the effect of oral consumption of <i>P. marinus</i> has not been investigated in humans or other mammals. To address the question we used humanized mice expressing HLA-DR4 molecules and lacking expression of mouse MHC-class II molecules (DR4.EA<sup>0</sup>) in such a way that CD4 T cell responses are solely restricted by the human HLA-DR4 molecule. The DR4.EA<sup>0</sup> mice did not develop diarrhea or any detectable pathology in the gastrointestinal tract or lungs following single or repeated feedings with live <i>P. marinus</i> parasites. Furthermore, lymphocyte populations in the gut associated lymphoid tissue and spleen were unaltered in the parasite-fed mice ruling out local or systemic inflammation. Notably, naïve DR4.EA<sup>0</sup> mice had antibodies (IgM and IgG) reacting against <i>P. marinus</i> parasites whereas parasite specific T cell responses were undetectable. Feeding with <i>P. marinus</i> boosted the antibody responses and stimulated specific cellular (IFNγ) immunity to the oyster parasite. Our data indicate the ability of <i>P. marinus</i> parasites to induce systemic immunity in DR4.EA<sup>0</sup> mice without causing noticeable pathology, and support rationale grounds for using genetically engineered <i>P. marinus</i> as a new oral vaccine platform to induce systemic immunity against infectious agents.</p></div>", "links"=>[], "tags"=>["immunology", "Immune cells", "Antibody-producing cells", "b cells", "t cells", "Immune system", "Lymphoid organs", "immunity", "Adaptive immunity", "Humoral immunity", "immunizations", "Immune response", "immunoglobulins", "Major histocompatibility complex", "Marine biology", "Fisheries science", "Model organisms", "Animal models", "mouse", "humanized", "hla-dr4", "mice", "fed", "protozoan", "pathogen", "oysters", "noticeable", "pathology", "elicit", "systemic"], "article_id"=>922458, "categories"=>["Biological Sciences"], "users"=>["Wathsala Wijayalath", "Sai Majji", "Yuliya Kleschenko", "Luis Pow-Sang", "Teodor D. Brumeanu", "Eileen Franke Villasante", "Gerardo R. Vasta", "José-Antonio Fernández-Robledo", "Sofia Casares"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087435", "stats"=>{"downloads"=>2, "page_views"=>160, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Humanized_HLA_DR4_Mice_Fed_with_the_Protozoan_Pathogen_of_Oysters_Perkinsus_Marinus_Dermo_Do_Not_Develop_Noticeable_Pathology_but_Elicit_Systemic_Immunity/922458", "title"=>"Humanized HLA-DR4 Mice Fed with the Protozoan Pathogen of Oysters <i>Perkinsus Marinus</i> (Dermo) Do Not Develop Noticeable Pathology but Elicit Systemic Immunity", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-31 03:38:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/1371707"], "description"=>"<p><b>Panel A,</b> groups DR4.EA<sup>0</sup> mice were fed with <i>P. marinus</i> twice at two-week interval and euthanized six days later. Controls were unfed mice. Spleen cells were stimulated with ConA (2 days) or <i>P. marinus</i> protein extract (4 days) and the levels of IFNγ in cell culture supernatants were measured by ELISA. Data represent mean ± SD of triplicate samples from three pooled spleens. <b>Panel B,</b> in an independent second experiment mice were fed with <i>P. marinus</i> as in panel A or unfed (control). Splenic cells were stimulated in triplicated samples (2 days) with <i>P. marinus</i> protein extracts or ConA and analyzed by ELISPOT. Control cultures were non-stimulated. Data represent mean spot forming units (sfu)/10<sup>6</sup> cells ± SD of triplicated samples from control (n = 3) and <i>P. marinus</i>-fed mice (n = 5) analyzed individually. T cells from control mice did not produce IFNγ upon stimulation with <i>P. marinus</i> protein extracts (p = 0.07, paired <i>t</i>-test) while T cells from <i>P. marinus</i>-fed responded to stimulation (p = 0.001, paired <i>t</i>-test). The T cell response to polyclonal stimulation with ConA was significantly higher in spleens of <i>P. marinus</i>-fed mice as compared to control mice (p = 0.0005, unpaired t-test). <b>Panel C&D,</b> Frequencies and numbers of B cells (CD19<sup>+</sup>), T cells (CD3<sup>+</sup>), CD3<sup>+</sup>CD4<sup>+</sup> and CD3<sup>+</sup>CD8<sup>+</sup> T cell subsets in spleens of mice fed once with <i>P. marinus</i> and examined at day 5 (1× d5) or day 14 (1× d14) post-feeding, mice fed twice with <i>P. marinus</i> (at two-week interval) and examined at day 6 post-feeding (2× d6) or control (unfed) mice. <b>Panel E,</b> Frequency of regulatory Foxp3<sup>+</sup> T cells in spleens on <i>P. marinus</i>-fed and control mice. Data in panels C–E represent mean ± SD of six mice fed with <i>P. marinus</i> and seven control (unfed) mice analyzed individually. NS, not significant (p>0.05) determined by unpaired <i>t</i>-test.</p>", "links"=>[], "tags"=>["immunology", "Immune cells", "Antibody-producing cells", "b cells", "t cells", "Immune system", "Lymphoid organs", "immunity", "Adaptive immunity", "Humoral immunity", "immunizations", "Immune response", "immunoglobulins", "Major histocompatibility complex", "Marine biology", "Fisheries science", "Model organisms", "Animal models", "mouse", "mice", "fed", "elicit", "cellular"], "article_id"=>922453, "categories"=>["Biological Sciences"], "users"=>["Wathsala Wijayalath", "Sai Majji", "Yuliya Kleschenko", "Luis Pow-Sang", "Teodor D. Brumeanu", "Eileen Franke Villasante", "Gerardo R. Vasta", "José-Antonio Fernández-Robledo", "Sofia Casares"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087435.g006", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DR4_EA_0_mice_fed_with_P_marinus_elicit_cellular_immunity_/922453", "title"=>"DR4.EA<sup>0</sup> mice fed with <i>P. marinus</i> elicit cellular immunity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-31 03:38:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/1371706"], "description"=>"<p><b>Panel A, </b><i>P. marinus</i> proteins extracted by thawing-freezing/ultrasonication as described in material and methods were separated in 4–15% SDS-PAGE gradient gel and silver-stained (lane 1); shown are the MW markers in lanes 2 (MagicMark) and 3 (Odyssey); <b>Panel B</b> show the same <i>P. marinus</i> protein sample after probing with sera from naïve mice (left) and from <i>P. marinus</i>-fed mice (right). The arrows indicate the major <i>P. marinus</i> protein bands recognized by IgG serum antibodies from naïve mice (arrows in panel B left, lane 1) and the most abundant <i>P. marinus</i> protein of approximately 60 kDa recognized by IgG serum antibodies from mice fed with <i>P. marinus</i> (arrow in panel B right, lane 1).</p>", "links"=>[], "tags"=>["immunology", "Immune cells", "Antibody-producing cells", "b cells", "t cells", "Immune system", "Lymphoid organs", "immunity", "Adaptive immunity", "Humoral immunity", "immunizations", "Immune response", "immunoglobulins", "Major histocompatibility complex", "Marine biology", "Fisheries science", "Model organisms", "Animal models", "mouse", "blot", "igg", "antibodies"], "article_id"=>922452, "categories"=>["Biological Sciences"], "users"=>["Wathsala Wijayalath", "Sai Majji", "Yuliya Kleschenko", "Luis Pow-Sang", "Teodor D. Brumeanu", "Eileen Franke Villasante", "Gerardo R. Vasta", "José-Antonio Fernández-Robledo", "Sofia Casares"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087435.g005", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Western_blot_analysis_of_IgG_antibodies_to_P_marinus_/922452", "title"=>"Western blot analysis of IgG antibodies to <i>P. marinus</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-31 03:38:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/1371703"], "description"=>"<p>DR4.EA<sup>0</sup> mice were fed by gavage with 10<sup>5</sup> live <i>P. marinus</i> and euthanized at 24 h, 48 h, or 7 days post-feeding (n = 3 mice per time point) for histological examination. Unfed age-matched mice were used as controls (n = 3). Organs fixed in formaline were processed and stained with H&E. Representative images were acquired using light microscopy under 10× objective (100× total visual magnification).</p>", "links"=>[], "tags"=>["immunology", "Immune cells", "Antibody-producing cells", "b cells", "t cells", "Immune system", "Lymphoid organs", "immunity", "Adaptive immunity", "Humoral immunity", "immunizations", "Immune response", "immunoglobulins", "Major histocompatibility complex", "Marine biology", "Fisheries science", "Model organisms", "Animal models", "mouse", "induce", "gastrointestinal", "pathology"], "article_id"=>922449, "categories"=>["Biological Sciences"], "users"=>["Wathsala Wijayalath", "Sai Majji", "Yuliya Kleschenko", "Luis Pow-Sang", "Teodor D. Brumeanu", "Eileen Franke Villasante", "Gerardo R. Vasta", "José-Antonio Fernández-Robledo", "Sofia Casares"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087435.g002", "stats"=>{"downloads"=>0, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Oral_administration_of_P_marinus_does_not_induce_gastrointestinal_or_lung_pathology_in_DR4_EA_0_mice_/922449", "title"=>"Oral administration of <i>P. marinus</i> does not induce gastrointestinal or lung pathology in DR4.EA<sup>0</sup> mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-31 03:38:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/1371708"], "description"=>"<p>PP, Peyer’s patches (PP); IEL, intraepithelial lymphocytes, LPL lamina propria lymphocytes. There were no significant differences between control (unfed) and <i>P. marinus</i>-fed mice (p>0.05 unpaired two-sample <i>t</i>-test).</p>", "links"=>[], "tags"=>["immunology", "Immune cells", "Antibody-producing cells", "b cells", "t cells", "Immune system", "Lymphoid organs", "immunity", "Adaptive immunity", "Humoral immunity", "immunizations", "Immune response", "immunoglobulins", "Major histocompatibility complex", "Marine biology", "Fisheries science", "Model organisms", "Animal models", "mouse", "cells"], "article_id"=>922454, "categories"=>["Biological Sciences"], "users"=>["Wathsala Wijayalath", "Sai Majji", "Yuliya Kleschenko", "Luis Pow-Sang", "Teodor D. Brumeanu", "Eileen Franke Villasante", "Gerardo R. Vasta", "José-Antonio Fernández-Robledo", "Sofia Casares"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087435.t001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Frequency_of_regulatory_CD4_Fop3_T_cells_Tregs_in_the_gut_/922454", "title"=>"Frequency of regulatory CD4<sup>+</sup>Fop3<sup>+</sup> T cells (Tregs) in the gut.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-01-31 03:38:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/1371705"], "description"=>"<p><b>Panel A,</b> IFA antibody titers in naïve (unfed) mice and mice fed with <i>P. marinus</i> (twice at two-week interval) measured at two weeks post-second feeding. Data represent titers in ten mice analyzed individually. <b>Panel B</b> shows representative IFAs.</p>", "links"=>[], "tags"=>["immunology", "Immune cells", "Antibody-producing cells", "b cells", "t cells", "Immune system", "Lymphoid organs", "immunity", "Adaptive immunity", "Humoral immunity", "immunizations", "Immune response", "immunoglobulins", "Major histocompatibility complex", "Marine biology", "Fisheries science", "Model organisms", "Animal models", "mouse", "mice", "elicit", "igg", "igm", "responses", "feeding"], "article_id"=>922451, "categories"=>["Biological Sciences"], "users"=>["Wathsala Wijayalath", "Sai Majji", "Yuliya Kleschenko", "Luis Pow-Sang", "Teodor D. Brumeanu", "Eileen Franke Villasante", "Gerardo R. Vasta", "José-Antonio Fernández-Robledo", "Sofia Casares"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087435.g004", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Humanized_mice_elicit_IgG_and_IgM_responses_following_oral_feeding_with_P_marinus_/922451", "title"=>"Humanized mice elicit IgG and IgM responses following oral feeding with <i>P. marinus</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-31 03:38:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/1371704"], "description"=>"<p>Groups of DR4.EA<sup>0</sup> mice were fed with <i>P. marinus</i> once and examined at days 5 or 14 after the feeding (n = 6 mice per time point) or they were fed twice (at two week interval) and examined at day 6 after the second feeding (n = 6). Controls (n = 7) were unfed mice. Lymphocytes isolated from Peyer’s patches (PP), intraepithelial (IEL), and lamina propria (PPL) were stained with mouse CD3, CD4, CD8, and CD19 Abs and analyzed by FACS. <b>Panel A,</b> frequency of B (CD19<sup>+</sup>) and T (CD3<sup>+</sup>) cells in control and <i>P. marinus</i>-fed mice. <b>Panel B,</b> frequency of CD4<sup>+</sup>, CD8<sup>+</sup>, CD4<sup>+</sup>CD8<sup>+</sup>, and CD4<sup>−</sup>CD8<sup>−</sup> T cell subsets among gated CD3<sup>+</sup> T cells. Data represent mean ± SD of mice analyzed individually. There were no significant differences for the frequency of lymphocytes in the gut of control mice as compared to mice fed with <i>P. marinus</i> (p>0.05 determine by unpaired <i>t</i>-test).</p>", "links"=>[], "tags"=>["immunology", "Immune cells", "Antibody-producing cells", "b cells", "t cells", "Immune system", "Lymphoid organs", "immunity", "Adaptive immunity", "Humoral immunity", "immunizations", "Immune response", "immunoglobulins", "Major histocompatibility complex", "Marine biology", "Fisheries science", "Model organisms", "Animal models", "mouse", "cells"], "article_id"=>922450, "categories"=>["Biological Sciences"], "users"=>["Wathsala Wijayalath", "Sai Majji", "Yuliya Kleschenko", "Luis Pow-Sang", "Teodor D. Brumeanu", "Eileen Franke Villasante", "Gerardo R. Vasta", "José-Antonio Fernández-Robledo", "Sofia Casares"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087435.g003", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_P_marinus_does_not_alter_the_frequency_of_T_and_B_cells_in_the_gut_/922450", "title"=>"<i>P. marinus</i> does not alter the frequency of T and B cells in the gut.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-31 03:38:15"}

PMC Usage Stats | Further Information

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