Comprehensive analysis of gene expression in the junctional epithelium by laser microdissection and microarray analysis
Hayashi Y, Matsunaga T, Yamamoto G, Nishii K, Usui M, Yamamoto M, Tachikawa T. Comprehensive analysis of gene expression in the junctional epithelium by laser microdissection and microarray analysis. J Periodont Res 2010; 45: 618–625. © 2010 John Wiley & Sons A/S Background and Objective: The j...
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| creator | Hayashi, Y. Matsunaga, T. Yamamoto, G. Nishii, K. Usui, M. Yamamoto, M. Tachikawa, T. |
| description | Hayashi Y, Matsunaga T, Yamamoto G, Nishii K, Usui M, Yamamoto M, Tachikawa T. Comprehensive analysis of gene expression in the junctional epithelium by laser microdissection and microarray analysis. J Periodont Res 2010; 45: 618–625. © 2010 John Wiley & Sons A/S
Background and Objective: The junctional epithelium attaches to the tooth enamel at the dentogingival junction. The attachment mechanisms of the junctional epithelium have been studied histologically, but the molecular functions of the junctional epithelium have not been elucidated.
The aim of this study was to perform a comprehensive analysis of gene expression in the junctional epithelium and to search for specific genetic markers of the junctional epithelium.
Material and Methods: A comprehensive analysis of genes expressed in the mouse junctional epithelium and oral gingival epithelium was performed using laser microdissection and microarray analysis. To extract high‐quality RNA from these tissues, we made frozen sections using a modified film method. Confirmation of the differential expression of selected genes was performed by quantitative real‐time PCR and immunohistochemistry.
Results: The modified method produced RNA of sufficient quality for microarray analysis. The result of microarray analysis showed that 841 genes were up‐regulated in the junctional epithelium compared with the oral gingival epithelium, and five were increased more than 50‐fold in the junctional epithelium. These five genes were secretory leukocyte protease inhibitor (Slpi), keratin 17 (Krt17), annexin A1 (Anxa1), myosin light peptide 6 (Myl6) and endoplasmic reticulum protein 29 (Erp29). In particular, Slpi expression in the junctional epithelium was approximately 100‐fold higher than in the oral gingival epithelium by real‐time PCR. Additionally, immunohistochemistry indicated that the Slpi protein is highly expressed in the junctional epithelium.
Conclusion: We developed a method for generating fresh‐frozen tissue sections suitable for extraction of good‐quality RNA. We determined that Slpi is characteristically expressed in the junctional epithelium. Our results provide a substantial advance in the analysis of gene expression in the junctional epithelium. |
| doi_str_mv | 10.1111/j.1600-0765.2010.01276.x |
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Background and Objective: The junctional epithelium attaches to the tooth enamel at the dentogingival junction. The attachment mechanisms of the junctional epithelium have been studied histologically, but the molecular functions of the junctional epithelium have not been elucidated.
The aim of this study was to perform a comprehensive analysis of gene expression in the junctional epithelium and to search for specific genetic markers of the junctional epithelium.
Material and Methods: A comprehensive analysis of genes expressed in the mouse junctional epithelium and oral gingival epithelium was performed using laser microdissection and microarray analysis. To extract high‐quality RNA from these tissues, we made frozen sections using a modified film method. Confirmation of the differential expression of selected genes was performed by quantitative real‐time PCR and immunohistochemistry.
Results: The modified method produced RNA of sufficient quality for microarray analysis. The result of microarray analysis showed that 841 genes were up‐regulated in the junctional epithelium compared with the oral gingival epithelium, and five were increased more than 50‐fold in the junctional epithelium. These five genes were secretory leukocyte protease inhibitor (Slpi), keratin 17 (Krt17), annexin A1 (Anxa1), myosin light peptide 6 (Myl6) and endoplasmic reticulum protein 29 (Erp29). In particular, Slpi expression in the junctional epithelium was approximately 100‐fold higher than in the oral gingival epithelium by real‐time PCR. Additionally, immunohistochemistry indicated that the Slpi protein is highly expressed in the junctional epithelium.
Conclusion: We developed a method for generating fresh‐frozen tissue sections suitable for extraction of good‐quality RNA. We determined that Slpi is characteristically expressed in the junctional epithelium. Our results provide a substantial advance in the analysis of gene expression in the junctional epithelium.</description><identifier>ISSN: 0022-3484</identifier><identifier>EISSN: 1600-0765</identifier><identifier>DOI: 10.1111/j.1600-0765.2010.01276.x</identifier><identifier>PMID: 20546111</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Animals ; Annexin A1 - biosynthesis ; Annexin A1 - genetics ; Antibiotics. Antiinfectious agents. Antiparasitic agents ; Antiviral agents ; Biological and medical sciences ; Dentistry ; Endoplasmic Reticulum ; Epithelial Attachment - enzymology ; Epithelial Attachment - metabolism ; Frozen Sections ; Gene Expression Profiling - methods ; Gingiva - metabolism ; Heat-Shock Proteins - biosynthesis ; Heat-Shock Proteins - genetics ; junctional epithelium ; Keratin-17 - biosynthesis ; Keratin-17 - genetics ; laser microdissection ; Lasers, Gas ; Medical sciences ; Mice ; microarray ; Microdissection - methods ; Myosin Light Chains - biosynthesis ; Myosin Light Chains - genetics ; Oligonucleotide Array Sequence Analysis ; Otorhinolaryngology. Stomatology ; Pharmacology. Drug treatments ; Secretory Leukocyte Peptidase Inhibitor - biosynthesis ; Secretory Leukocyte Peptidase Inhibitor - genetics ; secretory leukocyte protease inhibitor</subject><ispartof>Journal of periodontal research, 2010-10, Vol.45 (5), p.618-625</ispartof><rights>2010 John Wiley & Sons A/S</rights><rights>2015 INIST-CNRS</rights><rights>(c) 2010 John Wiley & Sons A/S.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4536-2331dda8df7a4ae3537b29e63a2f489f612068f30359fa4b0441fb567fccda7c3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1600-0765.2010.01276.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1600-0765.2010.01276.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>315,782,786,1419,27931,27932,45581,45582</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=23217124$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20546111$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hayashi, Y.</creatorcontrib><creatorcontrib>Matsunaga, T.</creatorcontrib><creatorcontrib>Yamamoto, G.</creatorcontrib><creatorcontrib>Nishii, K.</creatorcontrib><creatorcontrib>Usui, M.</creatorcontrib><creatorcontrib>Yamamoto, M.</creatorcontrib><creatorcontrib>Tachikawa, T.</creatorcontrib><title>Comprehensive analysis of gene expression in the junctional epithelium by laser microdissection and microarray analysis</title><title>Journal of periodontal research</title><addtitle>J Periodontal Res</addtitle><description>Hayashi Y, Matsunaga T, Yamamoto G, Nishii K, Usui M, Yamamoto M, Tachikawa T. Comprehensive analysis of gene expression in the junctional epithelium by laser microdissection and microarray analysis. J Periodont Res 2010; 45: 618–625. © 2010 John Wiley & Sons A/S
Background and Objective: The junctional epithelium attaches to the tooth enamel at the dentogingival junction. The attachment mechanisms of the junctional epithelium have been studied histologically, but the molecular functions of the junctional epithelium have not been elucidated.
The aim of this study was to perform a comprehensive analysis of gene expression in the junctional epithelium and to search for specific genetic markers of the junctional epithelium.
Material and Methods: A comprehensive analysis of genes expressed in the mouse junctional epithelium and oral gingival epithelium was performed using laser microdissection and microarray analysis. To extract high‐quality RNA from these tissues, we made frozen sections using a modified film method. Confirmation of the differential expression of selected genes was performed by quantitative real‐time PCR and immunohistochemistry.
Results: The modified method produced RNA of sufficient quality for microarray analysis. The result of microarray analysis showed that 841 genes were up‐regulated in the junctional epithelium compared with the oral gingival epithelium, and five were increased more than 50‐fold in the junctional epithelium. These five genes were secretory leukocyte protease inhibitor (Slpi), keratin 17 (Krt17), annexin A1 (Anxa1), myosin light peptide 6 (Myl6) and endoplasmic reticulum protein 29 (Erp29). In particular, Slpi expression in the junctional epithelium was approximately 100‐fold higher than in the oral gingival epithelium by real‐time PCR. Additionally, immunohistochemistry indicated that the Slpi protein is highly expressed in the junctional epithelium.
Conclusion: We developed a method for generating fresh‐frozen tissue sections suitable for extraction of good‐quality RNA. We determined that Slpi is characteristically expressed in the junctional epithelium. Our results provide a substantial advance in the analysis of gene expression in the junctional epithelium.</description><subject>Animals</subject><subject>Annexin A1 - biosynthesis</subject><subject>Annexin A1 - genetics</subject><subject>Antibiotics. Antiinfectious agents. Antiparasitic agents</subject><subject>Antiviral agents</subject><subject>Biological and medical sciences</subject><subject>Dentistry</subject><subject>Endoplasmic Reticulum</subject><subject>Epithelial Attachment - enzymology</subject><subject>Epithelial Attachment - metabolism</subject><subject>Frozen Sections</subject><subject>Gene Expression Profiling - methods</subject><subject>Gingiva - metabolism</subject><subject>Heat-Shock Proteins - biosynthesis</subject><subject>Heat-Shock Proteins - genetics</subject><subject>junctional epithelium</subject><subject>Keratin-17 - biosynthesis</subject><subject>Keratin-17 - genetics</subject><subject>laser microdissection</subject><subject>Lasers, Gas</subject><subject>Medical sciences</subject><subject>Mice</subject><subject>microarray</subject><subject>Microdissection - methods</subject><subject>Myosin Light Chains - biosynthesis</subject><subject>Myosin Light Chains - genetics</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>Otorhinolaryngology. Stomatology</subject><subject>Pharmacology. Drug treatments</subject><subject>Secretory Leukocyte Peptidase Inhibitor - biosynthesis</subject><subject>Secretory Leukocyte Peptidase Inhibitor - genetics</subject><subject>secretory leukocyte protease inhibitor</subject><issn>0022-3484</issn><issn>1600-0765</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkU1vEzEQhi1ERdPCX0C-IE4b_LV29sABotIWVUXiQ83Nmt0dU4f9CHa2zf57vE0afBnPzDPvSPMSQjmb8_Q-rOdcM5Yxo_O5YKnKuDB6vntBZsfGSzJjTIhMqoU6JWcxrlnKtSlekVPBcqWT0Iw8Lvt2E_Aeu-gfkEIHzRh9pL2jv7FDirvUjdH3HfUd3d4jXQ9dtU05NBQ3PlUaP7S0HGkDEQNtfRX62seIT1RSrPc1CAHG44LX5MRBE_HNIZ6TX18ufi6vsptvl9fLTzdZpXKpMyElr2tY1M6AApS5NKUoUEsQTi0Kp7lgeuEkk3nhQJVMKe7KXBtXVTWYSp6T93vdTej_Dhi3tvWxwqaBDvshWqOKdJWkm8i3B3IoW6ztJvgWwmifb5WAdwcAYgWNC9BVPv7npOCGC5W4j3vu0Tc4Hvuc2ck7u7aTRXayyE7e2Sfv7M5-_X4x_dJ8tp_3cYu74zyEP1YbaXJ7d3tpf9x9XqnV6jat_QepyZ4U</recordid><startdate>201010</startdate><enddate>201010</enddate><creator>Hayashi, Y.</creator><creator>Matsunaga, T.</creator><creator>Yamamoto, G.</creator><creator>Nishii, K.</creator><creator>Usui, M.</creator><creator>Yamamoto, M.</creator><creator>Tachikawa, T.</creator><general>Blackwell Publishing Ltd</general><general>Blackwell</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>201010</creationdate><title>Comprehensive analysis of gene expression in the junctional epithelium by laser microdissection and microarray analysis</title><author>Hayashi, Y. ; Matsunaga, T. ; Yamamoto, G. ; Nishii, K. ; Usui, M. ; Yamamoto, M. ; Tachikawa, T.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4536-2331dda8df7a4ae3537b29e63a2f489f612068f30359fa4b0441fb567fccda7c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Animals</topic><topic>Annexin A1 - biosynthesis</topic><topic>Annexin A1 - genetics</topic><topic>Antibiotics. Antiinfectious agents. Antiparasitic agents</topic><topic>Antiviral agents</topic><topic>Biological and medical sciences</topic><topic>Dentistry</topic><topic>Endoplasmic Reticulum</topic><topic>Epithelial Attachment - enzymology</topic><topic>Epithelial Attachment - metabolism</topic><topic>Frozen Sections</topic><topic>Gene Expression Profiling - methods</topic><topic>Gingiva - metabolism</topic><topic>Heat-Shock Proteins - biosynthesis</topic><topic>Heat-Shock Proteins - genetics</topic><topic>junctional epithelium</topic><topic>Keratin-17 - biosynthesis</topic><topic>Keratin-17 - genetics</topic><topic>laser microdissection</topic><topic>Lasers, Gas</topic><topic>Medical sciences</topic><topic>Mice</topic><topic>microarray</topic><topic>Microdissection - methods</topic><topic>Myosin Light Chains - biosynthesis</topic><topic>Myosin Light Chains - genetics</topic><topic>Oligonucleotide Array Sequence Analysis</topic><topic>Otorhinolaryngology. Stomatology</topic><topic>Pharmacology. Drug treatments</topic><topic>Secretory Leukocyte Peptidase Inhibitor - biosynthesis</topic><topic>Secretory Leukocyte Peptidase Inhibitor - genetics</topic><topic>secretory leukocyte protease inhibitor</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hayashi, Y.</creatorcontrib><creatorcontrib>Matsunaga, T.</creatorcontrib><creatorcontrib>Yamamoto, G.</creatorcontrib><creatorcontrib>Nishii, K.</creatorcontrib><creatorcontrib>Usui, M.</creatorcontrib><creatorcontrib>Yamamoto, M.</creatorcontrib><creatorcontrib>Tachikawa, T.</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of periodontal research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hayashi, Y.</au><au>Matsunaga, T.</au><au>Yamamoto, G.</au><au>Nishii, K.</au><au>Usui, M.</au><au>Yamamoto, M.</au><au>Tachikawa, T.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comprehensive analysis of gene expression in the junctional epithelium by laser microdissection and microarray analysis</atitle><jtitle>Journal of periodontal research</jtitle><addtitle>J Periodontal Res</addtitle><date>2010-10</date><risdate>2010</risdate><volume>45</volume><issue>5</issue><spage>618</spage><epage>625</epage><pages>618-625</pages><issn>0022-3484</issn><eissn>1600-0765</eissn><abstract>Hayashi Y, Matsunaga T, Yamamoto G, Nishii K, Usui M, Yamamoto M, Tachikawa T. Comprehensive analysis of gene expression in the junctional epithelium by laser microdissection and microarray analysis. J Periodont Res 2010; 45: 618–625. © 2010 John Wiley & Sons A/S
Background and Objective: The junctional epithelium attaches to the tooth enamel at the dentogingival junction. The attachment mechanisms of the junctional epithelium have been studied histologically, but the molecular functions of the junctional epithelium have not been elucidated.
The aim of this study was to perform a comprehensive analysis of gene expression in the junctional epithelium and to search for specific genetic markers of the junctional epithelium.
Material and Methods: A comprehensive analysis of genes expressed in the mouse junctional epithelium and oral gingival epithelium was performed using laser microdissection and microarray analysis. To extract high‐quality RNA from these tissues, we made frozen sections using a modified film method. Confirmation of the differential expression of selected genes was performed by quantitative real‐time PCR and immunohistochemistry.
Results: The modified method produced RNA of sufficient quality for microarray analysis. The result of microarray analysis showed that 841 genes were up‐regulated in the junctional epithelium compared with the oral gingival epithelium, and five were increased more than 50‐fold in the junctional epithelium. These five genes were secretory leukocyte protease inhibitor (Slpi), keratin 17 (Krt17), annexin A1 (Anxa1), myosin light peptide 6 (Myl6) and endoplasmic reticulum protein 29 (Erp29). In particular, Slpi expression in the junctional epithelium was approximately 100‐fold higher than in the oral gingival epithelium by real‐time PCR. Additionally, immunohistochemistry indicated that the Slpi protein is highly expressed in the junctional epithelium.
Conclusion: We developed a method for generating fresh‐frozen tissue sections suitable for extraction of good‐quality RNA. We determined that Slpi is characteristically expressed in the junctional epithelium. Our results provide a substantial advance in the analysis of gene expression in the junctional epithelium.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>20546111</pmid><doi>10.1111/j.1600-0765.2010.01276.x</doi><tpages>8</tpages></addata></record> |
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| subjects | Animals Annexin A1 - biosynthesis Annexin A1 - genetics Antibiotics. Antiinfectious agents. Antiparasitic agents Antiviral agents Biological and medical sciences Dentistry Endoplasmic Reticulum Epithelial Attachment - enzymology Epithelial Attachment - metabolism Frozen Sections Gene Expression Profiling - methods Gingiva - metabolism Heat-Shock Proteins - biosynthesis Heat-Shock Proteins - genetics junctional epithelium Keratin-17 - biosynthesis Keratin-17 - genetics laser microdissection Lasers, Gas Medical sciences Mice microarray Microdissection - methods Myosin Light Chains - biosynthesis Myosin Light Chains - genetics Oligonucleotide Array Sequence Analysis Otorhinolaryngology. Stomatology Pharmacology. Drug treatments Secretory Leukocyte Peptidase Inhibitor - biosynthesis Secretory Leukocyte Peptidase Inhibitor - genetics secretory leukocyte protease inhibitor |
| title | Comprehensive analysis of gene expression in the junctional epithelium by laser microdissection and microarray analysis |
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