The lymphocyte secretome from young adults enhances skeletal muscle proliferation and migration, but effects are attenuated in the secretome of older adults
Older people experience skeletal muscle wasting, in part due to impaired proliferative capacity of quiescent skeletal muscle satellite cells which can be reversed by exposure to young blood. To investigate the role of immune cells in muscle regeneration, we isolated lymphocytes from whole blood of y...
Gespeichert in:
| Veröffentlicht in: | Physiological reports 2015-11, Vol.3 (11), p.e12518-n/a |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | n/a |
|---|---|
| container_issue | 11 |
| container_start_page | e12518 |
| container_title | Physiological reports |
| container_volume | 3 |
| creator | Al‐Dabbagh, Sarah McPhee, Jamie S. Murgatroyd, Christopher Butler‐Browne, Gillian Stewart, Claire E. Al‐Shanti, Nasser |
| description | Older people experience skeletal muscle wasting, in part due to impaired proliferative capacity of quiescent skeletal muscle satellite cells which can be reversed by exposure to young blood. To investigate the role of immune cells in muscle regeneration, we isolated lymphocytes from whole blood of young and older healthy volunteers and cultured them with, or without, anti‐CD3/CD28 activators to induce release of cytokines, interleukins, and growth factors into the media. The secreted proteins were collected to prepare a conditioned media, which was subsequently used to culture C2C12 myoblasts. The conditioned media from the activated young lymphocytes increased the rate of proliferation of myoblasts by around threefold (P |
| doi_str_mv | 10.14814/phy2.12518 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4673618</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2289906420</sourcerecordid><originalsourceid>FETCH-LOGICAL-c4858-2b4af147e07aba5a594109d3fe773b8a8bbd4288111535477b5802d2261c32613</originalsourceid><addsrcrecordid>eNqNklFr1TAUx4s43Nj25LsEfBH0zpwkbdMXYQx1wmB7mKBPIU1PbzvTpCap0u_ih7Wu13H1QXw5OSE_ficJ_yx7CvQMhATxeuxmdgYsB_koO2I0h42E8tPjvf4wO43xjlIKlPOKiifZISsKyoWojrIftx0SOw9j582ckEQ0AZMfkLTBD2T2k9sS3Uw2RYKu085gJPELWkzakmGKxiIZg7d9i0Gn3juiXUOGfrvuXpF6SgTbFs1i0AGJTgndpBM2pHckdfszfUu8bTDsJp5kB622EU9363H28d3b24vLzdX1-w8X51cbI2QuN6wWugVRIi11rXOdVwJo1fAWy5LXUsu6bgSTEgBynouyrHNJWcNYAYYvhR9nb1bvONUDNgZdCtqqMfSDDrPyuld_nri-U1v_TYmi5AXIRfBiJwj-64QxqaGPBq3VDv0UFZQ5MOAVh_9ARVVQYIwv6PO_0Ds_Bbf8hGJMVhUtBKML9XKlTPAxBmwf7g1U3WdE_cqIus_IQj_bf-oD-zsRC8BW4Htvcf6XS91cfmar9SetecmB</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2289906420</pqid></control><display><type>article</type><title>The lymphocyte secretome from young adults enhances skeletal muscle proliferation and migration, but effects are attenuated in the secretome of older adults</title><source>DOAJ Directory of Open Access Journals</source><source>Wiley-Blackwell Open Access Titles</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Wiley Online Library All Journals</source><source>PubMed Central</source><creator>Al‐Dabbagh, Sarah ; McPhee, Jamie S. ; Murgatroyd, Christopher ; Butler‐Browne, Gillian ; Stewart, Claire E. ; Al‐Shanti, Nasser</creator><creatorcontrib>Al‐Dabbagh, Sarah ; McPhee, Jamie S. ; Murgatroyd, Christopher ; Butler‐Browne, Gillian ; Stewart, Claire E. ; Al‐Shanti, Nasser</creatorcontrib><description>Older people experience skeletal muscle wasting, in part due to impaired proliferative capacity of quiescent skeletal muscle satellite cells which can be reversed by exposure to young blood. To investigate the role of immune cells in muscle regeneration, we isolated lymphocytes from whole blood of young and older healthy volunteers and cultured them with, or without, anti‐CD3/CD28 activators to induce release of cytokines, interleukins, and growth factors into the media. The secreted proteins were collected to prepare a conditioned media, which was subsequently used to culture C2C12 myoblasts. The conditioned media from the activated young lymphocytes increased the rate of proliferation of myoblasts by around threefold (P < 0.005) and caused an approximate fourfold (P < 0.005) increase in migration compared with nonactivated lymphocyte control media. These responses were characterized by minimal myotube formation (2%), low fusion index (5%), low myosin heavy chain content, and substantial migration. In contrast, myoblasts treated with conditioned media from activated old lymphocytes exhibited a high degree of differentiation, and multi‐nucleated myotube formation that was comparable to control conditions, thus showing no effect on proliferation or migration of myoblasts. These results indicate that secreted proteins from lymphocytes of young people enhance the muscle cell proliferation and migration, whereas secreted proteins from lymphocytes of older people may contribute to the attenuated skeletal muscle satellite cell proliferation and migration.
Changes to the immune system in older age may contribute to impaired muscle regeneration and subsequent loss of skeletal muscle mass. Lymphocytes were collected from young and older adults. The proteins secreted by the lymphocytes after activation were harvested and added to skeletal muscle myoblasts in culture. The proteins secreted by activated lymphocytes from young improved myoblast proliferation by around three‐fold and myoblast migration by around four‐fold. The secreted by activated lymphocytes from older people had no effect on myoblasts. These results demonstrate that factors secreted from young lymphocytes are a mechanism for immune‐myoblast cell interactions and muscle regeneration, but the effects were impaired in lymphocytes from old people.</description><identifier>ISSN: 2051-817X</identifier><identifier>EISSN: 2051-817X</identifier><identifier>DOI: 10.14814/phy2.12518</identifier><identifier>PMID: 26603449</identifier><language>eng</language><publisher>United States: John Wiley & Sons, Inc</publisher><subject>Aging ; CD28 antigen ; CD3 antigen ; Cell culture ; Cell growth ; Cell proliferation ; Cytokines ; Differentiation ; Growth factors ; Immune system ; Interleukins ; Leukocyte migration ; Lymphocytes ; Media ; Musculoskeletal system ; Myoblasts ; Myosin ; Older people ; Original Research ; Penicillin ; Physiology ; Plasma ; proliferation ; Proteins ; Satellite cells ; Secretome ; Skeletal muscle ; Transplants & implants ; Trauma ; Young adults</subject><ispartof>Physiological reports, 2015-11, Vol.3 (11), p.e12518-n/a</ispartof><rights>2015 The Authors. published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.</rights><rights>2015 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.</rights><rights>2015. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2015 The Authors. published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society. 2015</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4858-2b4af147e07aba5a594109d3fe773b8a8bbd4288111535477b5802d2261c32613</citedby><cites>FETCH-LOGICAL-c4858-2b4af147e07aba5a594109d3fe773b8a8bbd4288111535477b5802d2261c32613</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4673618/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4673618/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,1417,11562,27924,27925,45574,45575,46052,46476,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26603449$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Al‐Dabbagh, Sarah</creatorcontrib><creatorcontrib>McPhee, Jamie S.</creatorcontrib><creatorcontrib>Murgatroyd, Christopher</creatorcontrib><creatorcontrib>Butler‐Browne, Gillian</creatorcontrib><creatorcontrib>Stewart, Claire E.</creatorcontrib><creatorcontrib>Al‐Shanti, Nasser</creatorcontrib><title>The lymphocyte secretome from young adults enhances skeletal muscle proliferation and migration, but effects are attenuated in the secretome of older adults</title><title>Physiological reports</title><addtitle>Physiol Rep</addtitle><description>Older people experience skeletal muscle wasting, in part due to impaired proliferative capacity of quiescent skeletal muscle satellite cells which can be reversed by exposure to young blood. To investigate the role of immune cells in muscle regeneration, we isolated lymphocytes from whole blood of young and older healthy volunteers and cultured them with, or without, anti‐CD3/CD28 activators to induce release of cytokines, interleukins, and growth factors into the media. The secreted proteins were collected to prepare a conditioned media, which was subsequently used to culture C2C12 myoblasts. The conditioned media from the activated young lymphocytes increased the rate of proliferation of myoblasts by around threefold (P < 0.005) and caused an approximate fourfold (P < 0.005) increase in migration compared with nonactivated lymphocyte control media. These responses were characterized by minimal myotube formation (2%), low fusion index (5%), low myosin heavy chain content, and substantial migration. In contrast, myoblasts treated with conditioned media from activated old lymphocytes exhibited a high degree of differentiation, and multi‐nucleated myotube formation that was comparable to control conditions, thus showing no effect on proliferation or migration of myoblasts. These results indicate that secreted proteins from lymphocytes of young people enhance the muscle cell proliferation and migration, whereas secreted proteins from lymphocytes of older people may contribute to the attenuated skeletal muscle satellite cell proliferation and migration.
Changes to the immune system in older age may contribute to impaired muscle regeneration and subsequent loss of skeletal muscle mass. Lymphocytes were collected from young and older adults. The proteins secreted by the lymphocytes after activation were harvested and added to skeletal muscle myoblasts in culture. The proteins secreted by activated lymphocytes from young improved myoblast proliferation by around three‐fold and myoblast migration by around four‐fold. The secreted by activated lymphocytes from older people had no effect on myoblasts. These results demonstrate that factors secreted from young lymphocytes are a mechanism for immune‐myoblast cell interactions and muscle regeneration, but the effects were impaired in lymphocytes from old people.</description><subject>Aging</subject><subject>CD28 antigen</subject><subject>CD3 antigen</subject><subject>Cell culture</subject><subject>Cell growth</subject><subject>Cell proliferation</subject><subject>Cytokines</subject><subject>Differentiation</subject><subject>Growth factors</subject><subject>Immune system</subject><subject>Interleukins</subject><subject>Leukocyte migration</subject><subject>Lymphocytes</subject><subject>Media</subject><subject>Musculoskeletal system</subject><subject>Myoblasts</subject><subject>Myosin</subject><subject>Older people</subject><subject>Original Research</subject><subject>Penicillin</subject><subject>Physiology</subject><subject>Plasma</subject><subject>proliferation</subject><subject>Proteins</subject><subject>Satellite cells</subject><subject>Secretome</subject><subject>Skeletal muscle</subject><subject>Transplants & implants</subject><subject>Trauma</subject><subject>Young adults</subject><issn>2051-817X</issn><issn>2051-817X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNklFr1TAUx4s43Nj25LsEfBH0zpwkbdMXYQx1wmB7mKBPIU1PbzvTpCap0u_ih7Wu13H1QXw5OSE_ficJ_yx7CvQMhATxeuxmdgYsB_koO2I0h42E8tPjvf4wO43xjlIKlPOKiifZISsKyoWojrIftx0SOw9j582ckEQ0AZMfkLTBD2T2k9sS3Uw2RYKu085gJPELWkzakmGKxiIZg7d9i0Gn3juiXUOGfrvuXpF6SgTbFs1i0AGJTgndpBM2pHckdfszfUu8bTDsJp5kB622EU9363H28d3b24vLzdX1-w8X51cbI2QuN6wWugVRIi11rXOdVwJo1fAWy5LXUsu6bgSTEgBynouyrHNJWcNYAYYvhR9nb1bvONUDNgZdCtqqMfSDDrPyuld_nri-U1v_TYmi5AXIRfBiJwj-64QxqaGPBq3VDv0UFZQ5MOAVh_9ARVVQYIwv6PO_0Ds_Bbf8hGJMVhUtBKML9XKlTPAxBmwf7g1U3WdE_cqIus_IQj_bf-oD-zsRC8BW4Htvcf6XS91cfmar9SetecmB</recordid><startdate>201511</startdate><enddate>201511</enddate><creator>Al‐Dabbagh, Sarah</creator><creator>McPhee, Jamie S.</creator><creator>Murgatroyd, Christopher</creator><creator>Butler‐Browne, Gillian</creator><creator>Stewart, Claire E.</creator><creator>Al‐Shanti, Nasser</creator><general>John Wiley & Sons, Inc</general><general>John Wiley & Sons, Ltd</general><scope>24P</scope><scope>WIN</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QP</scope><scope>7T5</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>201511</creationdate><title>The lymphocyte secretome from young adults enhances skeletal muscle proliferation and migration, but effects are attenuated in the secretome of older adults</title><author>Al‐Dabbagh, Sarah ; McPhee, Jamie S. ; Murgatroyd, Christopher ; Butler‐Browne, Gillian ; Stewart, Claire E. ; Al‐Shanti, Nasser</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4858-2b4af147e07aba5a594109d3fe773b8a8bbd4288111535477b5802d2261c32613</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Aging</topic><topic>CD28 antigen</topic><topic>CD3 antigen</topic><topic>Cell culture</topic><topic>Cell growth</topic><topic>Cell proliferation</topic><topic>Cytokines</topic><topic>Differentiation</topic><topic>Growth factors</topic><topic>Immune system</topic><topic>Interleukins</topic><topic>Leukocyte migration</topic><topic>Lymphocytes</topic><topic>Media</topic><topic>Musculoskeletal system</topic><topic>Myoblasts</topic><topic>Myosin</topic><topic>Older people</topic><topic>Original Research</topic><topic>Penicillin</topic><topic>Physiology</topic><topic>Plasma</topic><topic>proliferation</topic><topic>Proteins</topic><topic>Satellite cells</topic><topic>Secretome</topic><topic>Skeletal muscle</topic><topic>Transplants & implants</topic><topic>Trauma</topic><topic>Young adults</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Al‐Dabbagh, Sarah</creatorcontrib><creatorcontrib>McPhee, Jamie S.</creatorcontrib><creatorcontrib>Murgatroyd, Christopher</creatorcontrib><creatorcontrib>Butler‐Browne, Gillian</creatorcontrib><creatorcontrib>Stewart, Claire E.</creatorcontrib><creatorcontrib>Al‐Shanti, Nasser</creatorcontrib><collection>Wiley-Blackwell Open Access Titles</collection><collection>Wiley Free Content</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</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 Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Biological Science Database</collection><collection>Publicly Available Content 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>PubMed Central (Full Participant titles)</collection><jtitle>Physiological reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Al‐Dabbagh, Sarah</au><au>McPhee, Jamie S.</au><au>Murgatroyd, Christopher</au><au>Butler‐Browne, Gillian</au><au>Stewart, Claire E.</au><au>Al‐Shanti, Nasser</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The lymphocyte secretome from young adults enhances skeletal muscle proliferation and migration, but effects are attenuated in the secretome of older adults</atitle><jtitle>Physiological reports</jtitle><addtitle>Physiol Rep</addtitle><date>2015-11</date><risdate>2015</risdate><volume>3</volume><issue>11</issue><spage>e12518</spage><epage>n/a</epage><pages>e12518-n/a</pages><issn>2051-817X</issn><eissn>2051-817X</eissn><abstract>Older people experience skeletal muscle wasting, in part due to impaired proliferative capacity of quiescent skeletal muscle satellite cells which can be reversed by exposure to young blood. To investigate the role of immune cells in muscle regeneration, we isolated lymphocytes from whole blood of young and older healthy volunteers and cultured them with, or without, anti‐CD3/CD28 activators to induce release of cytokines, interleukins, and growth factors into the media. The secreted proteins were collected to prepare a conditioned media, which was subsequently used to culture C2C12 myoblasts. The conditioned media from the activated young lymphocytes increased the rate of proliferation of myoblasts by around threefold (P < 0.005) and caused an approximate fourfold (P < 0.005) increase in migration compared with nonactivated lymphocyte control media. These responses were characterized by minimal myotube formation (2%), low fusion index (5%), low myosin heavy chain content, and substantial migration. In contrast, myoblasts treated with conditioned media from activated old lymphocytes exhibited a high degree of differentiation, and multi‐nucleated myotube formation that was comparable to control conditions, thus showing no effect on proliferation or migration of myoblasts. These results indicate that secreted proteins from lymphocytes of young people enhance the muscle cell proliferation and migration, whereas secreted proteins from lymphocytes of older people may contribute to the attenuated skeletal muscle satellite cell proliferation and migration.
Changes to the immune system in older age may contribute to impaired muscle regeneration and subsequent loss of skeletal muscle mass. Lymphocytes were collected from young and older adults. The proteins secreted by the lymphocytes after activation were harvested and added to skeletal muscle myoblasts in culture. The proteins secreted by activated lymphocytes from young improved myoblast proliferation by around three‐fold and myoblast migration by around four‐fold. The secreted by activated lymphocytes from older people had no effect on myoblasts. These results demonstrate that factors secreted from young lymphocytes are a mechanism for immune‐myoblast cell interactions and muscle regeneration, but the effects were impaired in lymphocytes from old people.</abstract><cop>United States</cop><pub>John Wiley & Sons, Inc</pub><pmid>26603449</pmid><doi>10.14814/phy2.12518</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 2051-817X |
| ispartof | Physiological reports, 2015-11, Vol.3 (11), p.e12518-n/a |
| issn | 2051-817X 2051-817X |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4673618 |
| source | DOAJ Directory of Open Access Journals; Wiley-Blackwell Open Access Titles; EZB-FREE-00999 freely available EZB journals; Wiley Online Library All Journals; PubMed Central |
| subjects | Aging CD28 antigen CD3 antigen Cell culture Cell growth Cell proliferation Cytokines Differentiation Growth factors Immune system Interleukins Leukocyte migration Lymphocytes Media Musculoskeletal system Myoblasts Myosin Older people Original Research Penicillin Physiology Plasma proliferation Proteins Satellite cells Secretome Skeletal muscle Transplants & implants Trauma Young adults |
| title | The lymphocyte secretome from young adults enhances skeletal muscle proliferation and migration, but effects are attenuated in the secretome of older adults |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-07T22%3A16%3A13IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=The%20lymphocyte%20secretome%20from%20young%20adults%20enhances%20skeletal%20muscle%20proliferation%20and%20migration,%20but%20effects%20are%20attenuated%20in%20the%20secretome%20of%20older%20adults&rft.jtitle=Physiological%20reports&rft.au=Al%E2%80%90Dabbagh,%20Sarah&rft.date=2015-11&rft.volume=3&rft.issue=11&rft.spage=e12518&rft.epage=n/a&rft.pages=e12518-n/a&rft.issn=2051-817X&rft.eissn=2051-817X&rft_id=info:doi/10.14814/phy2.12518&rft_dat=%3Cproquest_pubme%3E2289906420%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2289906420&rft_id=info:pmid/26603449&rfr_iscdi=true |