mTORC1 inhibition with rapamycin exacerbates adipose tissue inflammation in obese mice and dissociates macrophage phenotype from function

Abstract Genetic- and diet-induced obesity and insulin resistance are associated with an increase in mechanistic target of rapamycin complex (mTORC) 1 activity in adipose tissue. We investigated herein the effects of pharmacological mTORC1 inhibition in the development of adipose tissue inflammation...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Immunobiology (1979) 2017-02, Vol.222 (2), p.261-271
Hauptverfasser: Paschoal, Vivian A, Amano, Mariane T, Belchior, Thiago, Magdalon, Juliana, Chimin, Patricia, Andrade, Maynara L, Ortiz-Silva, Milene, Castro, Érique, Yamashita, Alex S, Rosa Neto, José Cesar, Câmara, Niels O, Festuccia, William T
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 271
container_issue 2
container_start_page 261
container_title Immunobiology (1979)
container_volume 222
creator Paschoal, Vivian A
Amano, Mariane T
Belchior, Thiago
Magdalon, Juliana
Chimin, Patricia
Andrade, Maynara L
Ortiz-Silva, Milene
Castro, Érique
Yamashita, Alex S
Rosa Neto, José Cesar
Câmara, Niels O
Festuccia, William T
description Abstract Genetic- and diet-induced obesity and insulin resistance are associated with an increase in mechanistic target of rapamycin complex (mTORC) 1 activity in adipose tissue. We investigated herein the effects of pharmacological mTORC1 inhibition in the development of adipose tissue inflammation induced by high-fat diet (HFD) feeding, as well as in the polarization, metabolism and function of bone marrow-derived macrophages (BMDM). For this, C57BL/6J mice fed with a standard chow diet or a HFD (60% of calories from fat) and treated with either vehicle (0.1% Me2 SO, 0.2% methylcellulose) or rapamycin (2 mg/ kg/ day, gavage) during 30 days were evaluated for body weight, adiposity, glucose tolerance and adipose tissue inflammation. Although rapamycin did not affect the increase in body weight and adiposity, it exacerbated the glucose intolerance and adipose tissue inflammation induced by HFD feeding, as evidenced by the increased adipose tissue percentage of M1 macrophages, naive and activated cytotoxic T lymphocytes, and mRNA levels of proinflammatory molecules, such as TNF-α, IL-6 and MCP-1. In BMDM in vitro , pharmacological mTORC1 inhibition induced phosphorylation of NFκB p65 and spontaneous polarization of macrophages to a proinflammatory M1 profile, while it impaired M2 polarization induced by IL-4 + IL-13, glycolysis and phagocytosis. Altogether, these findings indicate that mTORC1 activity is an important determinant of adipose tissue inflammatory profile and macrophage plasticity, metabolism and function.
doi_str_mv 10.1016/j.imbio.2016.09.014
format Article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1835351799</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0171298516303813</els_id><sourcerecordid>1835351799</sourcerecordid><originalsourceid>FETCH-LOGICAL-c414t-8e0c43fdc35150d98b649adaf0accca759d57af15beacf6e926493084269f1573</originalsourceid><addsrcrecordid>eNqFks-OFCEQxonRuLOrT2BiOHrpFpr-x0ETM1HXZJNNdD2Taqh2GBtooVudR_CtZWZWD148Eaq-XxXUV4Q846zkjLcv96V1gw1llS8lkyXj9QOy4X3XF6Lq5EOyYbzjRSX75oJcprRnjMuq6x-Ti6prZY5XG_LL3d1-3HJq_c4OdrHB0x922dEIM7iDtp7iT9AYB1gwUTB2DgnpYlNaMUPjBM7BCcvSMGBOOquRgjfUZFXQ9kQ60DHMO_iCdN6hD8thRjrG4Oi4en0s8IQ8GmFK-PT-vCKf3729214XN7fvP2zf3BS65vVS9Mh0LUajRcMbZmQ_tLUEAyMDrTV0jTRNByNvBgQ9tiirnBesr6tW5mgnrsiLc905hm8rpkU5mzROE3gMa1K8F02u3UmZpeIszW9PKeKo5mgdxIPiTB09UHt18kAdPVBMquxBpp7fN1gHh-Yv82foWfDqLMD8ze8Wo0raotdobES9KBPsfxq8_ofXk_VWw_QVD5j2YY0-T1BxlSrF1KfjGhy3gLeCiZ4L8RsQ67Fj</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1835351799</pqid></control><display><type>article</type><title>mTORC1 inhibition with rapamycin exacerbates adipose tissue inflammation in obese mice and dissociates macrophage phenotype from function</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>Paschoal, Vivian A ; Amano, Mariane T ; Belchior, Thiago ; Magdalon, Juliana ; Chimin, Patricia ; Andrade, Maynara L ; Ortiz-Silva, Milene ; Castro, Érique ; Yamashita, Alex S ; Rosa Neto, José Cesar ; Câmara, Niels O ; Festuccia, William T</creator><creatorcontrib>Paschoal, Vivian A ; Amano, Mariane T ; Belchior, Thiago ; Magdalon, Juliana ; Chimin, Patricia ; Andrade, Maynara L ; Ortiz-Silva, Milene ; Castro, Érique ; Yamashita, Alex S ; Rosa Neto, José Cesar ; Câmara, Niels O ; Festuccia, William T</creatorcontrib><description>Abstract Genetic- and diet-induced obesity and insulin resistance are associated with an increase in mechanistic target of rapamycin complex (mTORC) 1 activity in adipose tissue. We investigated herein the effects of pharmacological mTORC1 inhibition in the development of adipose tissue inflammation induced by high-fat diet (HFD) feeding, as well as in the polarization, metabolism and function of bone marrow-derived macrophages (BMDM). For this, C57BL/6J mice fed with a standard chow diet or a HFD (60% of calories from fat) and treated with either vehicle (0.1% Me2 SO, 0.2% methylcellulose) or rapamycin (2 mg/ kg/ day, gavage) during 30 days were evaluated for body weight, adiposity, glucose tolerance and adipose tissue inflammation. Although rapamycin did not affect the increase in body weight and adiposity, it exacerbated the glucose intolerance and adipose tissue inflammation induced by HFD feeding, as evidenced by the increased adipose tissue percentage of M1 macrophages, naive and activated cytotoxic T lymphocytes, and mRNA levels of proinflammatory molecules, such as TNF-α, IL-6 and MCP-1. In BMDM in vitro , pharmacological mTORC1 inhibition induced phosphorylation of NFκB p65 and spontaneous polarization of macrophages to a proinflammatory M1 profile, while it impaired M2 polarization induced by IL-4 + IL-13, glycolysis and phagocytosis. Altogether, these findings indicate that mTORC1 activity is an important determinant of adipose tissue inflammatory profile and macrophage plasticity, metabolism and function.</description><identifier>ISSN: 0171-2985</identifier><identifier>EISSN: 1878-3279</identifier><identifier>DOI: 10.1016/j.imbio.2016.09.014</identifier><identifier>PMID: 27692982</identifier><language>eng</language><publisher>Netherlands: Elsevier GmbH</publisher><subject>Adipose tissue inflammation ; Advanced Basic Science ; Allergy and Immunology ; Animals ; Biomarkers ; Cytokines ; Cytokines - metabolism ; Glucose - metabolism ; Immunophenotyping ; Inflammation Mediators - metabolism ; Leukocytes - immunology ; Leukocytes - metabolism ; Leukocytes - pathology ; Macrophages ; Macrophages - drug effects ; Macrophages - immunology ; Macrophages - metabolism ; Macrophages, Peritoneal - drug effects ; Macrophages, Peritoneal - immunology ; Macrophages, Peritoneal - metabolism ; Mechanistic Target of Rapamycin Complex 1 - antagonists &amp; inhibitors ; Mice ; Mice, Inbred C57BL ; Mice, Obese ; mTOR ; Obesity ; Obesity - immunology ; Obesity - metabolism ; Obesity - pathology ; Panniculitis - immunology ; Panniculitis - metabolism ; Panniculitis - pathology ; Phenotype ; Rapamycin ; Sirolimus - pharmacology</subject><ispartof>Immunobiology (1979), 2017-02, Vol.222 (2), p.261-271</ispartof><rights>Elsevier GmbH</rights><rights>2016 Elsevier GmbH</rights><rights>Copyright © 2016 Elsevier GmbH. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c414t-8e0c43fdc35150d98b649adaf0accca759d57af15beacf6e926493084269f1573</citedby><cites>FETCH-LOGICAL-c414t-8e0c43fdc35150d98b649adaf0accca759d57af15beacf6e926493084269f1573</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.imbio.2016.09.014$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,782,786,3552,27931,27932,46002</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27692982$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Paschoal, Vivian A</creatorcontrib><creatorcontrib>Amano, Mariane T</creatorcontrib><creatorcontrib>Belchior, Thiago</creatorcontrib><creatorcontrib>Magdalon, Juliana</creatorcontrib><creatorcontrib>Chimin, Patricia</creatorcontrib><creatorcontrib>Andrade, Maynara L</creatorcontrib><creatorcontrib>Ortiz-Silva, Milene</creatorcontrib><creatorcontrib>Castro, Érique</creatorcontrib><creatorcontrib>Yamashita, Alex S</creatorcontrib><creatorcontrib>Rosa Neto, José Cesar</creatorcontrib><creatorcontrib>Câmara, Niels O</creatorcontrib><creatorcontrib>Festuccia, William T</creatorcontrib><title>mTORC1 inhibition with rapamycin exacerbates adipose tissue inflammation in obese mice and dissociates macrophage phenotype from function</title><title>Immunobiology (1979)</title><addtitle>Immunobiology</addtitle><description>Abstract Genetic- and diet-induced obesity and insulin resistance are associated with an increase in mechanistic target of rapamycin complex (mTORC) 1 activity in adipose tissue. We investigated herein the effects of pharmacological mTORC1 inhibition in the development of adipose tissue inflammation induced by high-fat diet (HFD) feeding, as well as in the polarization, metabolism and function of bone marrow-derived macrophages (BMDM). For this, C57BL/6J mice fed with a standard chow diet or a HFD (60% of calories from fat) and treated with either vehicle (0.1% Me2 SO, 0.2% methylcellulose) or rapamycin (2 mg/ kg/ day, gavage) during 30 days were evaluated for body weight, adiposity, glucose tolerance and adipose tissue inflammation. Although rapamycin did not affect the increase in body weight and adiposity, it exacerbated the glucose intolerance and adipose tissue inflammation induced by HFD feeding, as evidenced by the increased adipose tissue percentage of M1 macrophages, naive and activated cytotoxic T lymphocytes, and mRNA levels of proinflammatory molecules, such as TNF-α, IL-6 and MCP-1. In BMDM in vitro , pharmacological mTORC1 inhibition induced phosphorylation of NFκB p65 and spontaneous polarization of macrophages to a proinflammatory M1 profile, while it impaired M2 polarization induced by IL-4 + IL-13, glycolysis and phagocytosis. Altogether, these findings indicate that mTORC1 activity is an important determinant of adipose tissue inflammatory profile and macrophage plasticity, metabolism and function.</description><subject>Adipose tissue inflammation</subject><subject>Advanced Basic Science</subject><subject>Allergy and Immunology</subject><subject>Animals</subject><subject>Biomarkers</subject><subject>Cytokines</subject><subject>Cytokines - metabolism</subject><subject>Glucose - metabolism</subject><subject>Immunophenotyping</subject><subject>Inflammation Mediators - metabolism</subject><subject>Leukocytes - immunology</subject><subject>Leukocytes - metabolism</subject><subject>Leukocytes - pathology</subject><subject>Macrophages</subject><subject>Macrophages - drug effects</subject><subject>Macrophages - immunology</subject><subject>Macrophages - metabolism</subject><subject>Macrophages, Peritoneal - drug effects</subject><subject>Macrophages, Peritoneal - immunology</subject><subject>Macrophages, Peritoneal - metabolism</subject><subject>Mechanistic Target of Rapamycin Complex 1 - antagonists &amp; inhibitors</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Mice, Obese</subject><subject>mTOR</subject><subject>Obesity</subject><subject>Obesity - immunology</subject><subject>Obesity - metabolism</subject><subject>Obesity - pathology</subject><subject>Panniculitis - immunology</subject><subject>Panniculitis - metabolism</subject><subject>Panniculitis - pathology</subject><subject>Phenotype</subject><subject>Rapamycin</subject><subject>Sirolimus - pharmacology</subject><issn>0171-2985</issn><issn>1878-3279</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFks-OFCEQxonRuLOrT2BiOHrpFpr-x0ETM1HXZJNNdD2Taqh2GBtooVudR_CtZWZWD148Eaq-XxXUV4Q846zkjLcv96V1gw1llS8lkyXj9QOy4X3XF6Lq5EOyYbzjRSX75oJcprRnjMuq6x-Ti6prZY5XG_LL3d1-3HJq_c4OdrHB0x922dEIM7iDtp7iT9AYB1gwUTB2DgnpYlNaMUPjBM7BCcvSMGBOOquRgjfUZFXQ9kQ60DHMO_iCdN6hD8thRjrG4Oi4en0s8IQ8GmFK-PT-vCKf3729214XN7fvP2zf3BS65vVS9Mh0LUajRcMbZmQ_tLUEAyMDrTV0jTRNByNvBgQ9tiirnBesr6tW5mgnrsiLc905hm8rpkU5mzROE3gMa1K8F02u3UmZpeIszW9PKeKo5mgdxIPiTB09UHt18kAdPVBMquxBpp7fN1gHh-Yv82foWfDqLMD8ze8Wo0raotdobES9KBPsfxq8_ofXk_VWw_QVD5j2YY0-T1BxlSrF1KfjGhy3gLeCiZ4L8RsQ67Fj</recordid><startdate>20170201</startdate><enddate>20170201</enddate><creator>Paschoal, Vivian A</creator><creator>Amano, Mariane T</creator><creator>Belchior, Thiago</creator><creator>Magdalon, Juliana</creator><creator>Chimin, Patricia</creator><creator>Andrade, Maynara L</creator><creator>Ortiz-Silva, Milene</creator><creator>Castro, Érique</creator><creator>Yamashita, Alex S</creator><creator>Rosa Neto, José Cesar</creator><creator>Câmara, Niels O</creator><creator>Festuccia, William T</creator><general>Elsevier GmbH</general><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>7X8</scope></search><sort><creationdate>20170201</creationdate><title>mTORC1 inhibition with rapamycin exacerbates adipose tissue inflammation in obese mice and dissociates macrophage phenotype from function</title><author>Paschoal, Vivian A ; Amano, Mariane T ; Belchior, Thiago ; Magdalon, Juliana ; Chimin, Patricia ; Andrade, Maynara L ; Ortiz-Silva, Milene ; Castro, Érique ; Yamashita, Alex S ; Rosa Neto, José Cesar ; Câmara, Niels O ; Festuccia, William T</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c414t-8e0c43fdc35150d98b649adaf0accca759d57af15beacf6e926493084269f1573</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Adipose tissue inflammation</topic><topic>Advanced Basic Science</topic><topic>Allergy and Immunology</topic><topic>Animals</topic><topic>Biomarkers</topic><topic>Cytokines</topic><topic>Cytokines - metabolism</topic><topic>Glucose - metabolism</topic><topic>Immunophenotyping</topic><topic>Inflammation Mediators - metabolism</topic><topic>Leukocytes - immunology</topic><topic>Leukocytes - metabolism</topic><topic>Leukocytes - pathology</topic><topic>Macrophages</topic><topic>Macrophages - drug effects</topic><topic>Macrophages - immunology</topic><topic>Macrophages - metabolism</topic><topic>Macrophages, Peritoneal - drug effects</topic><topic>Macrophages, Peritoneal - immunology</topic><topic>Macrophages, Peritoneal - metabolism</topic><topic>Mechanistic Target of Rapamycin Complex 1 - antagonists &amp; inhibitors</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Mice, Obese</topic><topic>mTOR</topic><topic>Obesity</topic><topic>Obesity - immunology</topic><topic>Obesity - metabolism</topic><topic>Obesity - pathology</topic><topic>Panniculitis - immunology</topic><topic>Panniculitis - metabolism</topic><topic>Panniculitis - pathology</topic><topic>Phenotype</topic><topic>Rapamycin</topic><topic>Sirolimus - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Paschoal, Vivian A</creatorcontrib><creatorcontrib>Amano, Mariane T</creatorcontrib><creatorcontrib>Belchior, Thiago</creatorcontrib><creatorcontrib>Magdalon, Juliana</creatorcontrib><creatorcontrib>Chimin, Patricia</creatorcontrib><creatorcontrib>Andrade, Maynara L</creatorcontrib><creatorcontrib>Ortiz-Silva, Milene</creatorcontrib><creatorcontrib>Castro, Érique</creatorcontrib><creatorcontrib>Yamashita, Alex S</creatorcontrib><creatorcontrib>Rosa Neto, José Cesar</creatorcontrib><creatorcontrib>Câmara, Niels O</creatorcontrib><creatorcontrib>Festuccia, William T</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Immunobiology (1979)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Paschoal, Vivian A</au><au>Amano, Mariane T</au><au>Belchior, Thiago</au><au>Magdalon, Juliana</au><au>Chimin, Patricia</au><au>Andrade, Maynara L</au><au>Ortiz-Silva, Milene</au><au>Castro, Érique</au><au>Yamashita, Alex S</au><au>Rosa Neto, José Cesar</au><au>Câmara, Niels O</au><au>Festuccia, William T</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>mTORC1 inhibition with rapamycin exacerbates adipose tissue inflammation in obese mice and dissociates macrophage phenotype from function</atitle><jtitle>Immunobiology (1979)</jtitle><addtitle>Immunobiology</addtitle><date>2017-02-01</date><risdate>2017</risdate><volume>222</volume><issue>2</issue><spage>261</spage><epage>271</epage><pages>261-271</pages><issn>0171-2985</issn><eissn>1878-3279</eissn><abstract>Abstract Genetic- and diet-induced obesity and insulin resistance are associated with an increase in mechanistic target of rapamycin complex (mTORC) 1 activity in adipose tissue. We investigated herein the effects of pharmacological mTORC1 inhibition in the development of adipose tissue inflammation induced by high-fat diet (HFD) feeding, as well as in the polarization, metabolism and function of bone marrow-derived macrophages (BMDM). For this, C57BL/6J mice fed with a standard chow diet or a HFD (60% of calories from fat) and treated with either vehicle (0.1% Me2 SO, 0.2% methylcellulose) or rapamycin (2 mg/ kg/ day, gavage) during 30 days were evaluated for body weight, adiposity, glucose tolerance and adipose tissue inflammation. Although rapamycin did not affect the increase in body weight and adiposity, it exacerbated the glucose intolerance and adipose tissue inflammation induced by HFD feeding, as evidenced by the increased adipose tissue percentage of M1 macrophages, naive and activated cytotoxic T lymphocytes, and mRNA levels of proinflammatory molecules, such as TNF-α, IL-6 and MCP-1. In BMDM in vitro , pharmacological mTORC1 inhibition induced phosphorylation of NFκB p65 and spontaneous polarization of macrophages to a proinflammatory M1 profile, while it impaired M2 polarization induced by IL-4 + IL-13, glycolysis and phagocytosis. Altogether, these findings indicate that mTORC1 activity is an important determinant of adipose tissue inflammatory profile and macrophage plasticity, metabolism and function.</abstract><cop>Netherlands</cop><pub>Elsevier GmbH</pub><pmid>27692982</pmid><doi>10.1016/j.imbio.2016.09.014</doi><tpages>11</tpages></addata></record>
fulltext fulltext
identifier ISSN: 0171-2985
ispartof Immunobiology (1979), 2017-02, Vol.222 (2), p.261-271
issn 0171-2985
1878-3279
language eng
recordid cdi_proquest_miscellaneous_1835351799
source MEDLINE; Access via ScienceDirect (Elsevier)
subjects Adipose tissue inflammation
Advanced Basic Science
Allergy and Immunology
Animals
Biomarkers
Cytokines
Cytokines - metabolism
Glucose - metabolism
Immunophenotyping
Inflammation Mediators - metabolism
Leukocytes - immunology
Leukocytes - metabolism
Leukocytes - pathology
Macrophages
Macrophages - drug effects
Macrophages - immunology
Macrophages - metabolism
Macrophages, Peritoneal - drug effects
Macrophages, Peritoneal - immunology
Macrophages, Peritoneal - metabolism
Mechanistic Target of Rapamycin Complex 1 - antagonists & inhibitors
Mice
Mice, Inbred C57BL
Mice, Obese
mTOR
Obesity
Obesity - immunology
Obesity - metabolism
Obesity - pathology
Panniculitis - immunology
Panniculitis - metabolism
Panniculitis - pathology
Phenotype
Rapamycin
Sirolimus - pharmacology
title mTORC1 inhibition with rapamycin exacerbates adipose tissue inflammation in obese mice and dissociates macrophage phenotype from function
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-08T15%3A17%3A55IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=mTORC1%20inhibition%20with%20rapamycin%20exacerbates%20adipose%20tissue%20inflammation%20in%20obese%20mice%20and%20dissociates%20macrophage%20phenotype%20from%20function&rft.jtitle=Immunobiology%20(1979)&rft.au=Paschoal,%20Vivian%20A&rft.date=2017-02-01&rft.volume=222&rft.issue=2&rft.spage=261&rft.epage=271&rft.pages=261-271&rft.issn=0171-2985&rft.eissn=1878-3279&rft_id=info:doi/10.1016/j.imbio.2016.09.014&rft_dat=%3Cproquest_cross%3E1835351799%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1835351799&rft_id=info:pmid/27692982&rft_els_id=S0171298516303813&rfr_iscdi=true