Cleavage of Host Cytokeratin-6 by Lysine-Specific Gingipain Induces Gingival Inflammation in Periodontitis Patients
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{"title"=>"Cleavage of host cytokeratin-6 by lysine-specificgingipain induces gingival inflammation in periodontitis patients", "type"=>"journal", "authors"=>[{"first_name"=>"Salunya", "last_name"=>"Tancharoen", "scopus_author_id"=>"8357402700"}, {"first_name"=>"Takashi", "last_name"=>"Matsuyama", "scopus_author_id"=>"35311501600"}, {"first_name"=>"Ko Ichi", "last_name"=>"Kawahara", "scopus_author_id"=>"55684333700"}, {"first_name"=>"Kenji", "last_name"=>"Tanaka", "scopus_author_id"=>"55628544555"}, {"first_name"=>"Lyang Ja", "last_name"=>"Lee", "scopus_author_id"=>"7404389541"}, {"first_name"=>"Miho", "last_name"=>"Machigashira", "scopus_author_id"=>"6505779304"}, {"first_name"=>"Kazuyuki", "last_name"=>"Noguchi", "scopus_author_id"=>"56622177100"}, {"first_name"=>"Takashi", "last_name"=>"Ito", "scopus_author_id"=>"55554509800"}, {"first_name"=>"Takahisa", "last_name"=>"Imamura", "scopus_author_id"=>"7402482550"}, {"first_name"=>"Jan", "last_name"=>"Potempa", "scopus_author_id"=>"7102884964"}, {"first_name"=>"Kiyoshi", "last_name"=>"Kikuchi", "scopus_author_id"=>"55308440500"}, {"first_name"=>"Ikuro", "last_name"=>"Maruyama", "scopus_author_id"=>"55931098000"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"604166023", "sgr"=>"84928914572", "doi"=>"10.1371/journal.pone.0117775", "scopus"=>"2-s2.0-84928914572", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"25688865"}, "id"=>"00a35421-3783-3b10-bf12-2599c00ce260", "abstract"=>"BACKGROUND/PURPOSE: Lysine-specific gingipain (Kgp) is a virulence factor secreted from Porphyromonas gingivalis (P. gingivalis), a major etiological bacterium of periodontal disease. Keratin intermediate filaments maintain the structural integrity of gingival epithelial cells, but are targeted by Kgp to produce a novel cytokeratin 6 fragment (K6F). We investigated the release of K6F and its induction of cytokine secretion. METHODS: K6F present in the gingival crevicular fluid of periodontal disease patients and in gingipain-treated rat gingival epithelial cell culture supernatants was measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometer-based rapid quantitative peptide analysis using BLOTCHIP. K6F in gingival tissues was immunostained, and cytokeratin 6 protein was analyzed by immunofluorescence staining and flow cytometry. Activation of MAPK in gingival epithelial cells was evaluated by immunoblotting. ELISA was used to measure K6F and the cytokines release induced by K6F. Human gingival fibroblast migration was assessed using a Matrigel invasion chamber assay. RESULTS: We identified K6F, corresponding to the C-terminus region of human cytokeratin 6 (amino acids 359-378), in the gingival crevicular fluid of periodontal disease patients and in the supernatant from gingival epithelial cells cultured with Kgp. K6F antigen was distributed from the basal to the spinous epithelial layers in gingivae from periodontal disease patients. Cytokeratin 6 on gingival epithelial cells was degraded by Kgp, but not by Arg-gingipain, P. gingivalis lipopolysaccharide or Actinobacillus actinomycetemcomitans lipopolysaccharide. K6F, but not a scrambled K6F peptide, induced human gingival fibroblast migration and secretion of interleukin (IL)-6, IL-8 and monocyte chemoattractant protein-1. These effects of K6F were mediated by activation of p38 MAPK and Jun N-terminal kinase, but not p42/44 MAPK or p-Akt. CONCLUSION: Kgp degrades gingival epithelial cell cytokeratin 6 to K6F that, on release, induces invasion and cytokine secretion by human gingival fibroblasts. Thus, Kgp may contribute to the development of periodontal disease.", "link"=>"http://www.mendeley.com/research/cleavage-host-cytokeratin6-lysinespecificgingipain-induces-gingival-inflammation-periodontitis-patie", "reader_count"=>21, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Student > Doctoral Student"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>3, "Student > Postgraduate"=>3, "Student > Master"=>3, "Student > Bachelor"=>4, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Student > Doctoral Student"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>3, "Student > Postgraduate"=>3, "Student > Master"=>3, "Student > Bachelor"=>4, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>1, "Medicine and Dentistry"=>10, "Agricultural and Biological Sciences"=>8}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>10}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>8}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"United States"=>1}, "group_count"=>8}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1908658"], "description"=>"<p>A: Cells were either untreated (control, top), treated with 50 nM Kgp for 6 h or treated with Kgp inactivated by TLCK. After treatment, cells were stained with anti-cytokeratin 6 antibody (FITC, green) and Concanavalin A (ConA-Alexa 543, red), followed by DNA staining with DAPI (blue). Images were obtained by a fluorescence microscope at 400× magnification. Arrowhead indicates degraded cytokeratin-6 protein in rat GECs. Scale bar = 30 μm. B: Images were captured and fluorescence intensity was quantitated using Image J software. The percentage fluorescence intensity relative to control is shown and expressed as mean ± SD (n = 3). C: Flow cytometry of cytokeratin-6 protein is shown as the geometric mean of the fluorescence intensity (x-axis). The shaded histograms denote fluorescence in Kgp-treated cells before the assay. Data are representative of three independent experiments. *<i>P</i><0.05.</p>", "links"=>[], "tags"=>["p 38 MAPK", "K 6F Human gingival fibroblast migration", "Actinobacillus actinomycetemcomitans lipopolysaccharide", "cytokeratin 6 protein", "gingival fibroblast migration", "novel cytokeratin 6 fragment", "gingival crevicular fluid", "K 6F peptide", "K 6F", "k 6F antigen", "kgp", "disease patients", "elisa", "gingival epithelial cells", "Matrigel invasion chamber assay.ResultsWe", "spinous epithelial layers", "measure K 6F", "cytokine secretion.MethodsK 6F", "il", "BLOTCHIP"], "article_id"=>1310761, "categories"=>["Biological Sciences"], "users"=>["Salunya Tancharoen", "Takashi Matsuyama", "Ko-ichi Kawahara", "Kenji Tanaka", "Lyang-Ja Lee", "Miho Machigashira", "Kazuyuki Noguchi", "Takashi Ito", "Takahisa Imamura", "Jan Potempa", "Kiyoshi Kikuchi", "Ikuro Maruyama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117775.g004", "stats"=>{"downloads"=>1, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Rat_GEC_cytokeratin_6_degradation_by_Kgp_/1310761", "title"=>"Rat GEC cytokeratin-6 degradation by Kgp.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-17 02:57:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/1908650"], "description"=>"<p>A: Cytoskeletal protein was extracted from rat GECs treated with 10 nM Kgp, and cytokeratin-6 levels were measured by immunoblotting. Bands were quantified by densitometry. B: Cells were treated with <i>P</i>. <i>gingivalis</i> (<i>P</i>.<i>g</i>), <i>A</i>. <i>actinomycetemcomitans</i> (<i>A</i>.<i>a</i>) LPS or RgpB for 6 h before measurement of cytokeratin-6 levels in the cytoskeleton fraction. Vimentin was used as a loading control. Data are representative of three independent experiments. C: K6F in rat GEC supernatant after incubation with 10 nM Kgp as measured by ELISA. D: <i>P</i>.<i>g</i>., <i>A</i>.<i>a</i>. LPS or RgpB were used instead of Kgp. Data are the mean ± SD (n = 3).<i>*P<</i>0.05.</p>", "links"=>[], "tags"=>["p 38 MAPK", "K 6F Human gingival fibroblast migration", "Actinobacillus actinomycetemcomitans lipopolysaccharide", "cytokeratin 6 protein", "gingival fibroblast migration", "novel cytokeratin 6 fragment", "gingival crevicular fluid", "K 6F peptide", "K 6F", "k 6F antigen", "kgp", "disease patients", "elisa", "gingival epithelial cells", "Matrigel invasion chamber assay.ResultsWe", "spinous epithelial layers", "measure K 6F", "cytokine secretion.MethodsK 6F", "il", "BLOTCHIP"], "article_id"=>1310759, "categories"=>["Biological Sciences"], "users"=>["Salunya Tancharoen", "Takashi Matsuyama", "Ko-ichi Kawahara", "Kenji Tanaka", "Lyang-Ja Lee", "Miho Machigashira", "Kazuyuki Noguchi", "Takashi Ito", "Takahisa Imamura", "Jan Potempa", "Kiyoshi Kikuchi", "Ikuro Maruyama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117775.g003", "stats"=>{"downloads"=>0, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_Kgp_on_cytokeratin_6_in_the_cytoskeleton_fraction_and_K6F_in_rat_GEC_culture_supernatants_/1310759", "title"=>"Effects of Kgp on cytokeratin-6 in the cytoskeleton fraction and K6F in rat GEC culture supernatants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-17 02:57:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/1908649"], "description"=>"<p>Tissues from PD and healthy volunteers were snap-frozen and incubated with polyclonal antibody against (A) K6F or (B) ScK6F (5 μg/ml). C: Negative immunostaining for K6F in healthy and PD patients by isotype-matched control IgG. Arrow and arrowhead in Panel A(c) indicate K6F immunoreactivity on endothelial cells and the basal cell layer, respectively. Data are representative of three independent experiments. Scale bar, 50 micron.</p>", "links"=>[], "tags"=>["p 38 MAPK", "K 6F Human gingival fibroblast migration", "Actinobacillus actinomycetemcomitans lipopolysaccharide", "cytokeratin 6 protein", "gingival fibroblast migration", "novel cytokeratin 6 fragment", "gingival crevicular fluid", "K 6F peptide", "K 6F", "k 6F antigen", "kgp", "disease patients", "elisa", "gingival epithelial cells", "Matrigel invasion chamber assay.ResultsWe", "spinous epithelial layers", "measure K 6F", "cytokine secretion.MethodsK 6F", "il", "BLOTCHIP"], "article_id"=>1310758, "categories"=>["Biological Sciences"], "users"=>["Salunya Tancharoen", "Takashi Matsuyama", "Ko-ichi Kawahara", "Kenji Tanaka", "Lyang-Ja Lee", "Miho Machigashira", "Kazuyuki Noguchi", "Takashi Ito", "Takahisa Imamura", "Jan Potempa", "Kiyoshi Kikuchi", "Ikuro Maruyama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117775.g002", "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemistry_of_K6F_in_human_gingival_tissues_/1310758", "title"=>"Immunohistochemistry of K6F in human gingival tissues.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-17 02:57:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/1908664"], "description"=>"<p>A: Cells were seeded into the upper chamber of a transwell apparatus equipped with a 6.5-mm polycarbonate filter (8-μm pore) layered with 40 μg of Matrigel. Lower wells contained 500 μl of control medium supplemented with 1 μg/ml of K6F or ScK6F. The Matrigel chambers were incubated for 12 or 24 h in the presence or absence of 1 μM SB203580 or 1 μM SP600125 in the upper chamber. Cells in the top well were completely removed and cells on the underside of the membrane were counterstained with Mayer’s hematoxylin. B: Images were captured and cells were counted at ×40 magnification. The migration-enhancing effect is shown as the relative invasion capacity of the number of migratory cells in the sample versus that in control media. Data are the mean ± SD (n = 3) of two independent experiments. ‘NT’ denotes no treatment. <i>*P<</i>0.05 <i>vs</i>. control. <i>**P</i><0.05 <i>vs</i>. K6F alone.</p>", "links"=>[], "tags"=>["p 38 MAPK", "K 6F Human gingival fibroblast migration", "Actinobacillus actinomycetemcomitans lipopolysaccharide", "cytokeratin 6 protein", "gingival fibroblast migration", "novel cytokeratin 6 fragment", "gingival crevicular fluid", "K 6F peptide", "K 6F", "k 6F antigen", "kgp", "disease patients", "elisa", "gingival epithelial cells", "Matrigel invasion chamber assay.ResultsWe", "spinous epithelial layers", "measure K 6F", "cytokine secretion.MethodsK 6F", "il", "BLOTCHIP"], "article_id"=>1310767, "categories"=>["Biological Sciences"], "users"=>["Salunya Tancharoen", "Takashi Matsuyama", "Ko-ichi Kawahara", "Kenji Tanaka", "Lyang-Ja Lee", "Miho Machigashira", "Kazuyuki Noguchi", "Takashi Ito", "Takahisa Imamura", "Jan Potempa", "Kiyoshi Kikuchi", "Ikuro Maruyama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117775.g007", "stats"=>{"downloads"=>2, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_K6F_induced_HGF_migration_and_its_inhibition_by_p38_MAPK_and_JNK1_2_inhibitors_/1310767", "title"=>"K6F-induced HGF migration and its inhibition by p38 MAPK and JNK1/2 inhibitors.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-17 02:57:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/1908663"], "description"=>"<p>Cells were incubated with 1 μg/mL K6F or ScK6F before ELISA measurement of the levels of (A) MCP-1, (B) IL-6 and (C) IL-8 in the culture medium. D: Cells were incubated with 1 μg/mL K6F for 0–60 min, and the activation of p38 MAPK, JNK1/2, p42/p44 and Akt was determined by western blot analysis using antibodies that specifically recognize the activated forms of these kinases. Cells were pretreated with SB203580 (SB; a p38 inhibitor), SP600125 (SP; a JNK inhibitor) or U0126 (UO; a p42/p44 inhibitor) for 1 h, then incubated with K6F before measuring the levels of (E) MCP-1, (F) IL-6 and (G) IL-8in the culture medium by ELISA. Values denote mean ± SD (n = 3). <i>*P</i><0.05 <i>vs</i>. control.</p>", "links"=>[], "tags"=>["p 38 MAPK", "K 6F Human gingival fibroblast migration", "Actinobacillus actinomycetemcomitans lipopolysaccharide", "cytokeratin 6 protein", "gingival fibroblast migration", "novel cytokeratin 6 fragment", "gingival crevicular fluid", "K 6F peptide", "K 6F", "k 6F antigen", "kgp", "disease patients", "elisa", "gingival epithelial cells", "Matrigel invasion chamber assay.ResultsWe", "spinous epithelial layers", "measure K 6F", "cytokine secretion.MethodsK 6F", "il", "BLOTCHIP"], "article_id"=>1310766, "categories"=>["Biological Sciences"], "users"=>["Salunya Tancharoen", "Takashi Matsuyama", "Ko-ichi Kawahara", "Kenji Tanaka", "Lyang-Ja Lee", "Miho Machigashira", "Kazuyuki Noguchi", "Takashi Ito", "Takahisa Imamura", "Jan Potempa", "Kiyoshi Kikuchi", "Ikuro Maruyama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117775.g006", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_K6F_induced_cytokine_release_from_HGFs_/1310766", "title"=>"K6F-induced cytokine release from HGFs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-17 02:57:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/1908660"], "description"=>"<p>Cells were either untreated (control, top), or treated with FITC (green)-labeled K6F (middle) or FITC-labeled ScK6F antibody (bottom) (1 μg/ml). Cells were then counterstained with Alexa 543-labeled-ConA (red) and DAPI (blue). All images were obtained under an immunofluorescence microscope. Scale bar = 50 μm. Data are representative of three independent experiments.</p>", "links"=>[], "tags"=>["p 38 MAPK", "K 6F Human gingival fibroblast migration", "Actinobacillus actinomycetemcomitans lipopolysaccharide", "cytokeratin 6 protein", "gingival fibroblast migration", "novel cytokeratin 6 fragment", "gingival crevicular fluid", "K 6F peptide", "K 6F", "k 6F antigen", "kgp", "disease patients", "elisa", "gingival epithelial cells", "Matrigel invasion chamber assay.ResultsWe", "spinous epithelial layers", "measure K 6F", "cytokine secretion.MethodsK 6F", "il", "BLOTCHIP"], "article_id"=>1310763, "categories"=>["Biological Sciences"], "users"=>["Salunya Tancharoen", "Takashi Matsuyama", "Ko-ichi Kawahara", "Kenji Tanaka", "Lyang-Ja Lee", "Miho Machigashira", "Kazuyuki Noguchi", "Takashi Ito", "Takahisa Imamura", "Jan Potempa", "Kiyoshi Kikuchi", "Ikuro Maruyama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117775.g005", "stats"=>{"downloads"=>3, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Presence_of_K6F_on_human_gingival_fibroblasts_/1310763", "title"=>"Presence of K6F on human gingival fibroblasts.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-17 02:57:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/1908687", "https://ndownloader.figshare.com/files/1908688", "https://ndownloader.figshare.com/files/1908689"], "description"=>"<div><p>Background/Purpose</p><p>Lysine-specific gingipain (Kgp) is a virulence factor secreted from Porphyromonas gingivalis (P. gingivalis), a major etiological bacterium of periodontal disease. Keratin intermediate filaments maintain the structural integrity of gingival epithelial cells, but are targeted by Kgp to produce a novel cytokeratin 6 fragment (K6F). We investigated the release of K6F and its induction of cytokine secretion.</p><p>Methods</p><p>K6F present in the gingival crevicular fluid of periodontal disease patients and in gingipain-treated rat gingival epithelial cell culture supernatants was measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometer-based rapid quantitative peptide analysis using BLOTCHIP. K6F in gingival tissues was immunostained, and cytokeratin 6 protein was analyzed by immunofluorescence staining and flow cytometry. Activation of MAPK in gingival epithelial cells was evaluated by immunoblotting. ELISA was used to measure K6F and the cytokines release induced by K6F. Human gingival fibroblast migration was assessed using a Matrigel invasion chamber assay.</p><p>Results</p><p>We identified K6F, corresponding to the C-terminus region of human cytokeratin 6 (amino acids 359–378), in the gingival crevicular fluid of periodontal disease patients and in the supernatant from gingival epithelial cells cultured with Kgp. K6F antigen was distributed from the basal to the spinous epithelial layers in gingivae from periodontal disease patients. Cytokeratin 6 on gingival epithelial cells was degraded by Kgp, but not by Arg-gingipain, P. gingivalis lipopolysaccharide or Actinobacillus actinomycetemcomitans lipopolysaccharide. K6F, but not a scrambled K6F peptide, induced human gingival fibroblast migration and secretion of interleukin (IL)-6, IL-8 and monocyte chemoattractant protein-1. These effects of K6F were mediated by activation of p38 MAPK and Jun N-terminal kinase, but not p42/44 MAPK or p-Akt.</p><p>Conclusion</p><p>Kgp degrades gingival epithelial cell cytokeratin 6 to K6F that, on release, induces invasion and cytokine secretion by human gingival fibroblasts. Thus, Kgp may contribute to the development of periodontal disease.</p></div>", "links"=>[], "tags"=>["p 38 MAPK", "K 6F Human gingival fibroblast migration", "Actinobacillus actinomycetemcomitans lipopolysaccharide", "cytokeratin 6 protein", "gingival fibroblast migration", "novel cytokeratin 6 fragment", "gingival crevicular fluid", "K 6F peptide", "K 6F", "k 6F antigen", "kgp", "disease patients", "elisa", "gingival epithelial cells", "Matrigel invasion chamber assay.ResultsWe", "spinous epithelial layers", "measure K 6F", "cytokine secretion.MethodsK 6F", "il", "BLOTCHIP"], "article_id"=>1310780, "categories"=>["Biological Sciences"], "users"=>["Salunya Tancharoen", "Takashi Matsuyama", "Ko-ichi Kawahara", "Kenji Tanaka", "Lyang-Ja Lee", "Miho Machigashira", "Kazuyuki Noguchi", "Takashi Ito", "Takahisa Imamura", "Jan Potempa", "Kiyoshi Kikuchi", "Ikuro Maruyama"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0117775.s001", "https://dx.doi.org/10.1371/journal.pone.0117775.s002", "https://dx.doi.org/10.1371/journal.pone.0117775.s003"], "stats"=>{"downloads"=>5, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cleavage_of_Host_Cytokeratin_6_by_Lysine_Specific_Gingipain_Induces_Gingival_Inflammation_in_Periodontitis_Patients_/1310780", "title"=>"Cleavage of Host Cytokeratin-6 by Lysine-Specific Gingipain Induces Gingival Inflammation in Periodontitis Patients", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-02-17 02:57:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/1908642"], "description"=>"<p>A: The proteins in GCF were extracted from the strips into the assay buffer and analyzed by MALDI-TOF mass spectrometry in arbitrary units (a.u.). The profile of GCF from PD patients on K6F peptides (red line) was superimposed onto that of healthy subjects (blue line). B: Profile of K6F detected in rat GEC supernatants. Activated gingipains were incubated with primary cultures of rat GECs, and supernatants were collected. A peak of K6F was detected in supernatants from rat GECs treated with Kgp (red line) at a molecular weight of 2217. C: GCF samples were collected and K6F titers were quantitatively measured by ELISA. Chart shows a dot plot of K6F levels in individual patients. Experiments were performed two times in triplicate. *<i>P</i><0.001 versus PD.</p>", "links"=>[], "tags"=>["p 38 MAPK", "K 6F Human gingival fibroblast migration", "Actinobacillus actinomycetemcomitans lipopolysaccharide", "cytokeratin 6 protein", "gingival fibroblast migration", "novel cytokeratin 6 fragment", "gingival crevicular fluid", "K 6F peptide", "K 6F", "k 6F antigen", "kgp", "disease patients", "elisa", "gingival epithelial cells", "Matrigel invasion chamber assay.ResultsWe", "spinous epithelial layers", "measure K 6F", "cytokine secretion.MethodsK 6F", "il", "BLOTCHIP"], "article_id"=>1310751, "categories"=>["Biological Sciences"], "users"=>["Salunya Tancharoen", "Takashi Matsuyama", "Ko-ichi Kawahara", "Kenji Tanaka", "Lyang-Ja Lee", "Miho Machigashira", "Kazuyuki Noguchi", "Takashi Ito", "Takahisa Imamura", "Jan Potempa", "Kiyoshi Kikuchi", "Ikuro Maruyama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0117775.g001", "stats"=>{"downloads"=>0, "page_views"=>41, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MALDI_TOF_mass_spectrometry_profiles_of_GCF_and_rat_GEC_culture_supernatants_and_K6F_concentrations_in_GCF_from_PD_patients_and_healthy_volunteers_/1310751", "title"=>"MALDI-TOF mass spectrometry profiles of GCF and rat GEC culture supernatants and K6F concentrations in GCF from PD patients and healthy volunteers.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-02-17 02:57:30"}

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  • {"unique-ip"=>"15", "full-text"=>"18", "pdf"=>"7", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2018", "month"=>"5"}
  • {"unique-ip"=>"8", "full-text"=>"4", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"2", "cited-by"=>"1", "year"=>"2018", "month"=>"6"}
  • {"unique-ip"=>"13", "full-text"=>"9", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"2", "cited-by"=>"1", "year"=>"2018", "month"=>"7"}
  • {"unique-ip"=>"9", "full-text"=>"7", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2018", "month"=>"8"}
  • {"unique-ip"=>"10", "full-text"=>"12", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2018", "month"=>"10"}
  • {"unique-ip"=>"17", "full-text"=>"18", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2018", "month"=>"11"}
  • {"unique-ip"=>"61", "full-text"=>"59", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"9", "full-text"=>"6", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"10", "full-text"=>"6", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"1", "cited-by"=>"1", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"6", "full-text"=>"5", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"14", "full-text"=>"12", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"7", "full-text"=>"7", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2019", "month"=>"10"}
  • {"unique-ip"=>"8", "full-text"=>"4", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"2", "cited-by"=>"1", "year"=>"2019", "month"=>"12"}
  • {"unique-ip"=>"6", "full-text"=>"3", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"4", "cited-by"=>"1", "year"=>"2020", "month"=>"2"}
  • {"unique-ip"=>"12", "full-text"=>"10", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"5", "cited-by"=>"0", "year"=>"2020", "month"=>"3"}
  • {"unique-ip"=>"3", "full-text"=>"1", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2020", "month"=>"4"}
  • {"unique-ip"=>"8", "full-text"=>"8", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"5"}
  • {"unique-ip"=>"9", "full-text"=>"9", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"6"}
  • {"unique-ip"=>"11", "full-text"=>"9", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2020", "month"=>"7"}
  • {"unique-ip"=>"7", "full-text"=>"11", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"8"}
  • {"unique-ip"=>"12", "full-text"=>"10", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"9"}
  • {"unique-ip"=>"13", "full-text"=>"10", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"5", "cited-by"=>"0", "year"=>"2020", "month"=>"10"}

Relative Metric

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