Rebeccamycin Attenuates TNF-α-Induced Intestinal Epithelial Barrier Dysfunction by Inhibiting Myosin Light Chain Kinase Production
Background/Aims: Although proinflammatory cytokine–induced disruption of intestinal epithelial barrier integrity is associated with intestinal inflammatory disease, effective treatment for barrier dysfunction is lacking. Previously, we demonstrated that rebeccamycin alleviates epithelial barrier dys...
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| Veröffentlicht in: | Cellular physiology and biochemistry 2017-01, Vol.41 (5), p.1924-1934 |
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| container_title | Cellular physiology and biochemistry |
| container_volume | 41 |
| creator | Watari, Akihiro Sakamoto, Yuta Hisaie, Kota Iwamoto, Kazuki Fueta, Miho Yagi, Kiyohito Kondoh, Masuo |
| description | Background/Aims: Although proinflammatory cytokine–induced disruption of intestinal epithelial barrier integrity is associated with intestinal inflammatory disease, effective treatment for barrier dysfunction is lacking. Previously, we demonstrated that rebeccamycin alleviates epithelial barrier dysfunction induced by inflammatory cytokines in Caco-2 cell monolayers; however, the underlying mechanism remained unclear. Here, we investigated the mechanism by which rebeccamycin protects the epithelial barrier function of Caco-2 cells exposed to TNF-α. Methods: To confirm the epithelial barrier function of Caco-2 cell monolayers, transepithelial electrical resistance (TER) and paracellular permeability were measured. Production levels and localization of tight junction (TJ) proteins were analyzed by immunoblot and immunofluorescence, respectively. Phosphorylated myosin light chain (pMLC) and MLC kinase (MLCK) mRNA expression levels were determined by immunoblot and quantitative RT-PCR, respectively. Results: Rebeccamycin attenuated the TNF-α-induced reduction in TER and increase in paracellular permeability. Rebeccamycin increased claudin-5 expression, but not claudin-1, -2, -4, occludin or ZO-1 expression, and prevented the TNF-α-induced changes in ZO-1 and occludin localization. Rebeccamycin suppressed the TNF-α-induced increase in MLCK mRNA expression, thus suppressing MLC phosphorylation. The rebeccamycin-mediated reduction in MLCK production and protection of epithelial barrier function were alleviated by Chk1 inhibition. Conclusion: Rebeccamycin attenuates TNF-α-induced disruption of intestinal epithelial barrier integrity by inducing claudin-5 expression and suppressing MLCK production via Chk1 activation. |
| doi_str_mv | 10.1159/000472367 |
| format | Article |
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Previously, we demonstrated that rebeccamycin alleviates epithelial barrier dysfunction induced by inflammatory cytokines in Caco-2 cell monolayers; however, the underlying mechanism remained unclear. Here, we investigated the mechanism by which rebeccamycin protects the epithelial barrier function of Caco-2 cells exposed to TNF-α. Methods: To confirm the epithelial barrier function of Caco-2 cell monolayers, transepithelial electrical resistance (TER) and paracellular permeability were measured. Production levels and localization of tight junction (TJ) proteins were analyzed by immunoblot and immunofluorescence, respectively. Phosphorylated myosin light chain (pMLC) and MLC kinase (MLCK) mRNA expression levels were determined by immunoblot and quantitative RT-PCR, respectively. Results: Rebeccamycin attenuated the TNF-α-induced reduction in TER and increase in paracellular permeability. Rebeccamycin increased claudin-5 expression, but not claudin-1, -2, -4, occludin or ZO-1 expression, and prevented the TNF-α-induced changes in ZO-1 and occludin localization. Rebeccamycin suppressed the TNF-α-induced increase in MLCK mRNA expression, thus suppressing MLC phosphorylation. The rebeccamycin-mediated reduction in MLCK production and protection of epithelial barrier function were alleviated by Chk1 inhibition. Conclusion: Rebeccamycin attenuates TNF-α-induced disruption of intestinal epithelial barrier integrity by inducing claudin-5 expression and suppressing MLCK production via Chk1 activation.</description><identifier>ISSN: 1015-8987</identifier><identifier>EISSN: 1421-9778</identifier><identifier>DOI: 10.1159/000472367</identifier><identifier>PMID: 28391269</identifier><language>eng</language><publisher>Basel, Switzerland: S. Karger AG</publisher><subject>Caco-2 Cells ; Carbazoles - pharmacology ; Checkpoint kinase 1 ; Checkpoint Kinase 1 - metabolism ; Claudin-5 - biosynthesis ; Enzyme Activation - drug effects ; Epithelial barrier ; Gene Expression Regulation, Enzymologic - drug effects ; Humans ; Inflammatory bowel disease ; Intestinal cells ; Intestinal Mucosa - enzymology ; Kinases ; Myosin light chain kinase ; Myosin Light Chains - metabolism ; Myosin-Light-Chain Kinase - biosynthesis ; Original Paper ; Phosphorylation - drug effects ; Small intestine ; Tight junction ; Tight Junctions - enzymology ; TNF-α ; Tumor Necrosis Factor-alpha - pharmacology ; Tumor necrosis factor-TNF</subject><ispartof>Cellular physiology and biochemistry, 2017-01, Vol.41 (5), p.1924-1934</ispartof><rights>2017 The Author(s)Published by S. Karger AG, Basel</rights><rights>2017 The Author(s)Published by S. Karger AG, Basel.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c463t-6d553dcf19c1eabcf044e935e37283a52404797d9ca38eeae8f81c1585e8c3573</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,860,2096,27614,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28391269$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Watari, Akihiro</creatorcontrib><creatorcontrib>Sakamoto, Yuta</creatorcontrib><creatorcontrib>Hisaie, Kota</creatorcontrib><creatorcontrib>Iwamoto, Kazuki</creatorcontrib><creatorcontrib>Fueta, Miho</creatorcontrib><creatorcontrib>Yagi, Kiyohito</creatorcontrib><creatorcontrib>Kondoh, Masuo</creatorcontrib><title>Rebeccamycin Attenuates TNF-α-Induced Intestinal Epithelial Barrier Dysfunction by Inhibiting Myosin Light Chain Kinase Production</title><title>Cellular physiology and biochemistry</title><addtitle>Cell Physiol Biochem</addtitle><description>Background/Aims: Although proinflammatory cytokine–induced disruption of intestinal epithelial barrier integrity is associated with intestinal inflammatory disease, effective treatment for barrier dysfunction is lacking. Previously, we demonstrated that rebeccamycin alleviates epithelial barrier dysfunction induced by inflammatory cytokines in Caco-2 cell monolayers; however, the underlying mechanism remained unclear. Here, we investigated the mechanism by which rebeccamycin protects the epithelial barrier function of Caco-2 cells exposed to TNF-α. Methods: To confirm the epithelial barrier function of Caco-2 cell monolayers, transepithelial electrical resistance (TER) and paracellular permeability were measured. Production levels and localization of tight junction (TJ) proteins were analyzed by immunoblot and immunofluorescence, respectively. Phosphorylated myosin light chain (pMLC) and MLC kinase (MLCK) mRNA expression levels were determined by immunoblot and quantitative RT-PCR, respectively. Results: Rebeccamycin attenuated the TNF-α-induced reduction in TER and increase in paracellular permeability. Rebeccamycin increased claudin-5 expression, but not claudin-1, -2, -4, occludin or ZO-1 expression, and prevented the TNF-α-induced changes in ZO-1 and occludin localization. Rebeccamycin suppressed the TNF-α-induced increase in MLCK mRNA expression, thus suppressing MLC phosphorylation. The rebeccamycin-mediated reduction in MLCK production and protection of epithelial barrier function were alleviated by Chk1 inhibition. Conclusion: Rebeccamycin attenuates TNF-α-induced disruption of intestinal epithelial barrier integrity by inducing claudin-5 expression and suppressing MLCK production via Chk1 activation.</description><subject>Caco-2 Cells</subject><subject>Carbazoles - pharmacology</subject><subject>Checkpoint kinase 1</subject><subject>Checkpoint Kinase 1 - metabolism</subject><subject>Claudin-5 - biosynthesis</subject><subject>Enzyme Activation - drug effects</subject><subject>Epithelial barrier</subject><subject>Gene Expression Regulation, Enzymologic - drug effects</subject><subject>Humans</subject><subject>Inflammatory bowel disease</subject><subject>Intestinal cells</subject><subject>Intestinal Mucosa - enzymology</subject><subject>Kinases</subject><subject>Myosin light chain kinase</subject><subject>Myosin Light Chains - metabolism</subject><subject>Myosin-Light-Chain Kinase - biosynthesis</subject><subject>Original Paper</subject><subject>Phosphorylation - drug effects</subject><subject>Small intestine</subject><subject>Tight junction</subject><subject>Tight Junctions - enzymology</subject><subject>TNF-α</subject><subject>Tumor Necrosis Factor-alpha - pharmacology</subject><subject>Tumor necrosis factor-TNF</subject><issn>1015-8987</issn><issn>1421-9778</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>M--</sourceid><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNptkcFuEzEQhlcIREvhwB2hlbjAYcFee9f2sQ0tRASoUDlbXnucOGzWqb17yJkn4kV4JqYkBAlx8tj6_I09f1E8peQ1pY16QwjhomatuFecUl7TSgkh72NNaFNJJcVJ8SjnNcGtUPXD4qSWTNG6VafF9y_QgbVms7NhKM_HEYbJjJDLm09X1c8f1XxwkwVXzgc8HMNg-vJyG8YV9AHLC5NSgFS-3WU_DXYMcSi7HcKr0AWkl-XHXcwoXoTlaixnK4P1B7RkKK9TRPXdlcfFA2_6DE8O61nx9eryZva-Wnx-N5-dLyrLWzZWrWsa5qynylIwnfWEc1CsASbwP6apOU5BCaesYRLAgPSSWtrIBqRljWBnxXzvddGs9TaFjUk7HU3Qvw9iWmqTxmB70NYo2TrjhO8k945I3lmiwHai81wagq6Xe9c2xdsJR6M3IVvoezNAnLKmUraMt4QpRF_8g67jlHCSSClFJG0xO6Re7SmbYs4J_PGBlOi7lPUxZWSfH4xTtwF3JP_E-rflN5OWkI7A7Ppir9Bb55F69l_q0OUXwo24bg</recordid><startdate>20170101</startdate><enddate>20170101</enddate><creator>Watari, Akihiro</creator><creator>Sakamoto, Yuta</creator><creator>Hisaie, Kota</creator><creator>Iwamoto, Kazuki</creator><creator>Fueta, Miho</creator><creator>Yagi, Kiyohito</creator><creator>Kondoh, Masuo</creator><general>S. Karger AG</general><general>Cell Physiol Biochem Press GmbH & Co KG</general><scope>M--</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>DOA</scope></search><sort><creationdate>20170101</creationdate><title>Rebeccamycin Attenuates TNF-α-Induced Intestinal Epithelial Barrier Dysfunction by Inhibiting Myosin Light Chain Kinase Production</title><author>Watari, Akihiro ; Sakamoto, Yuta ; Hisaie, Kota ; Iwamoto, Kazuki ; Fueta, Miho ; Yagi, Kiyohito ; Kondoh, Masuo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c463t-6d553dcf19c1eabcf044e935e37283a52404797d9ca38eeae8f81c1585e8c3573</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Caco-2 Cells</topic><topic>Carbazoles - pharmacology</topic><topic>Checkpoint kinase 1</topic><topic>Checkpoint Kinase 1 - metabolism</topic><topic>Claudin-5 - biosynthesis</topic><topic>Enzyme Activation - drug effects</topic><topic>Epithelial barrier</topic><topic>Gene Expression Regulation, Enzymologic - drug effects</topic><topic>Humans</topic><topic>Inflammatory bowel disease</topic><topic>Intestinal cells</topic><topic>Intestinal Mucosa - enzymology</topic><topic>Kinases</topic><topic>Myosin light chain kinase</topic><topic>Myosin Light Chains - metabolism</topic><topic>Myosin-Light-Chain Kinase - biosynthesis</topic><topic>Original Paper</topic><topic>Phosphorylation - drug effects</topic><topic>Small intestine</topic><topic>Tight junction</topic><topic>Tight Junctions - enzymology</topic><topic>TNF-α</topic><topic>Tumor Necrosis Factor-alpha - pharmacology</topic><topic>Tumor necrosis factor-TNF</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Watari, Akihiro</creatorcontrib><creatorcontrib>Sakamoto, Yuta</creatorcontrib><creatorcontrib>Hisaie, Kota</creatorcontrib><creatorcontrib>Iwamoto, Kazuki</creatorcontrib><creatorcontrib>Fueta, Miho</creatorcontrib><creatorcontrib>Yagi, Kiyohito</creatorcontrib><creatorcontrib>Kondoh, Masuo</creatorcontrib><collection>Karger Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>Cellular physiology and biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Watari, Akihiro</au><au>Sakamoto, Yuta</au><au>Hisaie, Kota</au><au>Iwamoto, Kazuki</au><au>Fueta, Miho</au><au>Yagi, Kiyohito</au><au>Kondoh, Masuo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rebeccamycin Attenuates TNF-α-Induced Intestinal Epithelial Barrier Dysfunction by Inhibiting Myosin Light Chain Kinase Production</atitle><jtitle>Cellular physiology and biochemistry</jtitle><addtitle>Cell Physiol Biochem</addtitle><date>2017-01-01</date><risdate>2017</risdate><volume>41</volume><issue>5</issue><spage>1924</spage><epage>1934</epage><pages>1924-1934</pages><issn>1015-8987</issn><eissn>1421-9778</eissn><abstract>Background/Aims: Although proinflammatory cytokine–induced disruption of intestinal epithelial barrier integrity is associated with intestinal inflammatory disease, effective treatment for barrier dysfunction is lacking. Previously, we demonstrated that rebeccamycin alleviates epithelial barrier dysfunction induced by inflammatory cytokines in Caco-2 cell monolayers; however, the underlying mechanism remained unclear. Here, we investigated the mechanism by which rebeccamycin protects the epithelial barrier function of Caco-2 cells exposed to TNF-α. Methods: To confirm the epithelial barrier function of Caco-2 cell monolayers, transepithelial electrical resistance (TER) and paracellular permeability were measured. Production levels and localization of tight junction (TJ) proteins were analyzed by immunoblot and immunofluorescence, respectively. Phosphorylated myosin light chain (pMLC) and MLC kinase (MLCK) mRNA expression levels were determined by immunoblot and quantitative RT-PCR, respectively. Results: Rebeccamycin attenuated the TNF-α-induced reduction in TER and increase in paracellular permeability. Rebeccamycin increased claudin-5 expression, but not claudin-1, -2, -4, occludin or ZO-1 expression, and prevented the TNF-α-induced changes in ZO-1 and occludin localization. Rebeccamycin suppressed the TNF-α-induced increase in MLCK mRNA expression, thus suppressing MLC phosphorylation. The rebeccamycin-mediated reduction in MLCK production and protection of epithelial barrier function were alleviated by Chk1 inhibition. Conclusion: Rebeccamycin attenuates TNF-α-induced disruption of intestinal epithelial barrier integrity by inducing claudin-5 expression and suppressing MLCK production via Chk1 activation.</abstract><cop>Basel, Switzerland</cop><pub>S. Karger AG</pub><pmid>28391269</pmid><doi>10.1159/000472367</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Caco-2 Cells Carbazoles - pharmacology Checkpoint kinase 1 Checkpoint Kinase 1 - metabolism Claudin-5 - biosynthesis Enzyme Activation - drug effects Epithelial barrier Gene Expression Regulation, Enzymologic - drug effects Humans Inflammatory bowel disease Intestinal cells Intestinal Mucosa - enzymology Kinases Myosin light chain kinase Myosin Light Chains - metabolism Myosin-Light-Chain Kinase - biosynthesis Original Paper Phosphorylation - drug effects Small intestine Tight junction Tight Junctions - enzymology TNF-α Tumor Necrosis Factor-alpha - pharmacology Tumor necrosis factor-TNF |
| title | Rebeccamycin Attenuates TNF-α-Induced Intestinal Epithelial Barrier Dysfunction by Inhibiting Myosin Light Chain Kinase Production |
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