Biochemical, pharmacological and structural characterization of two PLA2 isoforms Cdr-12 and Cdr-13 from Crotalus durissus ruruima snake venom
Cdr-12 and Cdr-13 isoforms of PLA2, a D49 protein, were purified from Crotalus durissus ruruima venom after one chromatographic step, reverse phase HPLC on micro-Bondapack C-18. The molecular mass by SDS-PAGE of Cdr-12 and Cdr-13 isoforms of PLA2 was 14333.49 Da and 14296.42 Da, respectively and con...
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| Veröffentlicht in: | The Protein Journal 2007-01, Vol.26 (1), p.39-49 |
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| creator | Ponce-Soto, Luis Alberto Baldasso, Paulo Aparecido Romero-Vargas, Frey Francisco Winck, Flávia V Novello, José Camillo Marangoni, Sergio |
| description | Cdr-12 and Cdr-13 isoforms of PLA2, a D49 protein, were purified from Crotalus durissus ruruima venom after one chromatographic step, reverse phase HPLC on micro-Bondapack C-18. The molecular mass by SDS-PAGE of Cdr-12 and Cdr-13 isoforms of PLA2 was 14333.49 Da and 14296.42 Da, respectively and confirmed by MALDI-TOF mass spectrometry. The amino acid composition showed that both isoforms Cdr-12 and Cdr-13 have a high content of Lys, Tyr, Gly, Arg, and 14 half-Cys residues, typical of a basic PLA2. The isoforms Cdr-12 and Cdr-13 had a sequence of amino acids of 122 amino acid residues, being Cdr-12: SLLQFNKMIK FETRKNAIPF YAFYGCYCGW GGQGRPKDAT DRCCIVHDCC YGKLAKCNTK WDFYRYSLRS GYFQCGKGTW CEQQICECDR VAAECLRRSL STYRYGYMIY PDSRCREPSE TC and pI value 8.37 and Cdr-13: SLVQFEKMIK EETGKNAVPF YAFYGCYCGW GGRGRPKDAT DRCCIVHDCC YEKLVKCNTK WDFYRYSLRS GYFQCGKGTW CEQQICECDR VAAECLRRSL STYRYGKMIY PDSRCREPSE TC with a pI value of 8.13 This sequence shows high identity values when compared to other D49 PLA2s isolated from venoms of crotalics snakes. Skeletal muscle preparations from the young chicken have been previously used in order to study the effects of toxins on neuromuscular transmission, providing an important opportunity to study the differentiated behavior of a toxin before more than one model, because it shows differences in its sensibilities. In mice, the PLA2 isoforms Cdr-12 and Cdr-13 induced myonecrosis and edema, upon intramuscular or subcutaneous injections, respectively. In vitro, Cdr-12 and Cdr-13 isoforms of PLA2, caused a potent blockade of neuromuscular transmission in young chicken biventer cervicis preparation and produced cytotoxicity in murine C2C12 skeletal muscle myotubes and lack cytolytic activity upon myoblasts in vitro. Thus, the combined structural and functional information obtained identify Cdr-12 and Cdr-13 isoforms as members of the PLA2 family, which presents the typical bioactivities described for such proteins. |
| doi_str_mv | 10.1007/s10930-006-9042-3 |
| format | Article |
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The molecular mass by SDS-PAGE of Cdr-12 and Cdr-13 isoforms of PLA2 was 14333.49 Da and 14296.42 Da, respectively and confirmed by MALDI-TOF mass spectrometry. The amino acid composition showed that both isoforms Cdr-12 and Cdr-13 have a high content of Lys, Tyr, Gly, Arg, and 14 half-Cys residues, typical of a basic PLA2. The isoforms Cdr-12 and Cdr-13 had a sequence of amino acids of 122 amino acid residues, being Cdr-12: SLLQFNKMIK FETRKNAIPF YAFYGCYCGW GGQGRPKDAT DRCCIVHDCC YGKLAKCNTK WDFYRYSLRS GYFQCGKGTW CEQQICECDR VAAECLRRSL STYRYGYMIY PDSRCREPSE TC and pI value 8.37 and Cdr-13: SLVQFEKMIK EETGKNAVPF YAFYGCYCGW GGRGRPKDAT DRCCIVHDCC YEKLVKCNTK WDFYRYSLRS GYFQCGKGTW CEQQICECDR VAAECLRRSL STYRYGKMIY PDSRCREPSE TC with a pI value of 8.13 This sequence shows high identity values when compared to other D49 PLA2s isolated from venoms of crotalics snakes. Skeletal muscle preparations from the young chicken have been previously used in order to study the effects of toxins on neuromuscular transmission, providing an important opportunity to study the differentiated behavior of a toxin before more than one model, because it shows differences in its sensibilities. In mice, the PLA2 isoforms Cdr-12 and Cdr-13 induced myonecrosis and edema, upon intramuscular or subcutaneous injections, respectively. In vitro, Cdr-12 and Cdr-13 isoforms of PLA2, caused a potent blockade of neuromuscular transmission in young chicken biventer cervicis preparation and produced cytotoxicity in murine C2C12 skeletal muscle myotubes and lack cytolytic activity upon myoblasts in vitro. Thus, the combined structural and functional information obtained identify Cdr-12 and Cdr-13 isoforms as members of the PLA2 family, which presents the typical bioactivities described for such proteins.</description><identifier>ISSN: 1572-3887</identifier><identifier>EISSN: 1875-8355</identifier><identifier>EISSN: 1573-4943</identifier><identifier>DOI: 10.1007/s10930-006-9042-3</identifier><identifier>PMID: 17203396</identifier><language>eng</language><publisher>Netherlands: Springer Nature B.V</publisher><subject>Amino acid composition ; Amino Acid Sequence ; Amino acids ; Animals ; Biocompatibility ; Cells, Cultured ; Chemical bonds ; Chemical Fractionation ; Chickens ; Crotalid Venoms - chemistry ; Crotalid Venoms - enzymology ; Crotalid Venoms - isolation & purification ; Crotalid Venoms - toxicity ; Crotalus ; Crotalus durissus ruruima ; Cytolytic activity ; Cytotoxicity ; Diaphragm - drug effects ; Edema ; Gel electrophoresis ; High performance liquid chromatography ; Isoelectric Point ; Isoenzymes ; Isoforms ; Liquid chromatography ; Mass spectrometry ; Mass spectroscopy ; Mice ; Molecular Weight ; Muscle, Skeletal - pathology ; Muscles ; Myoblasts ; Myoblasts - drug effects ; Myoblasts - metabolism ; Myonecrosis ; Myotubes ; Necrosis - pathology ; Neuromuscular junctions ; Neuromuscular transmission ; Phospholipase A2 ; Phospholipases A - chemistry ; Phospholipases A - isolation & purification ; Phospholipases A - toxicity ; Phospholipases A2 ; Phrenic Nerve - drug effects ; Proteins ; Residues ; Sequence Alignment ; Skeletal muscle ; Snakes ; Sodium lauryl sulfate ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Structural analysis ; Structure-function relationships ; Toxicity ; Toxins ; Venom</subject><ispartof>The Protein Journal, 2007-01, Vol.26 (1), p.39-49</ispartof><rights>Springer Science+Business Media, LLC 2007.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c241t-90418fd7eebaac5d5e6a3331e3ae290b35107783a349fc8eb44810767d8a6a863</citedby><cites>FETCH-LOGICAL-c241t-90418fd7eebaac5d5e6a3331e3ae290b35107783a349fc8eb44810767d8a6a863</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17203396$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ponce-Soto, Luis Alberto</creatorcontrib><creatorcontrib>Baldasso, Paulo Aparecido</creatorcontrib><creatorcontrib>Romero-Vargas, Frey Francisco</creatorcontrib><creatorcontrib>Winck, Flávia V</creatorcontrib><creatorcontrib>Novello, José Camillo</creatorcontrib><creatorcontrib>Marangoni, Sergio</creatorcontrib><title>Biochemical, pharmacological and structural characterization of two PLA2 isoforms Cdr-12 and Cdr-13 from Crotalus durissus ruruima snake venom</title><title>The Protein Journal</title><addtitle>Protein J</addtitle><description>Cdr-12 and Cdr-13 isoforms of PLA2, a D49 protein, were purified from Crotalus durissus ruruima venom after one chromatographic step, reverse phase HPLC on micro-Bondapack C-18. The molecular mass by SDS-PAGE of Cdr-12 and Cdr-13 isoforms of PLA2 was 14333.49 Da and 14296.42 Da, respectively and confirmed by MALDI-TOF mass spectrometry. The amino acid composition showed that both isoforms Cdr-12 and Cdr-13 have a high content of Lys, Tyr, Gly, Arg, and 14 half-Cys residues, typical of a basic PLA2. The isoforms Cdr-12 and Cdr-13 had a sequence of amino acids of 122 amino acid residues, being Cdr-12: SLLQFNKMIK FETRKNAIPF YAFYGCYCGW GGQGRPKDAT DRCCIVHDCC YGKLAKCNTK WDFYRYSLRS GYFQCGKGTW CEQQICECDR VAAECLRRSL STYRYGYMIY PDSRCREPSE TC and pI value 8.37 and Cdr-13: SLVQFEKMIK EETGKNAVPF YAFYGCYCGW GGRGRPKDAT DRCCIVHDCC YEKLVKCNTK WDFYRYSLRS GYFQCGKGTW CEQQICECDR VAAECLRRSL STYRYGKMIY PDSRCREPSE TC with a pI value of 8.13 This sequence shows high identity values when compared to other D49 PLA2s isolated from venoms of crotalics snakes. Skeletal muscle preparations from the young chicken have been previously used in order to study the effects of toxins on neuromuscular transmission, providing an important opportunity to study the differentiated behavior of a toxin before more than one model, because it shows differences in its sensibilities. In mice, the PLA2 isoforms Cdr-12 and Cdr-13 induced myonecrosis and edema, upon intramuscular or subcutaneous injections, respectively. In vitro, Cdr-12 and Cdr-13 isoforms of PLA2, caused a potent blockade of neuromuscular transmission in young chicken biventer cervicis preparation and produced cytotoxicity in murine C2C12 skeletal muscle myotubes and lack cytolytic activity upon myoblasts in vitro. Thus, the combined structural and functional information obtained identify Cdr-12 and Cdr-13 isoforms as members of the PLA2 family, which presents the typical bioactivities described for such proteins.</description><subject>Amino acid composition</subject><subject>Amino Acid Sequence</subject><subject>Amino acids</subject><subject>Animals</subject><subject>Biocompatibility</subject><subject>Cells, Cultured</subject><subject>Chemical bonds</subject><subject>Chemical Fractionation</subject><subject>Chickens</subject><subject>Crotalid Venoms - chemistry</subject><subject>Crotalid Venoms - enzymology</subject><subject>Crotalid Venoms - isolation & purification</subject><subject>Crotalid Venoms - toxicity</subject><subject>Crotalus</subject><subject>Crotalus durissus ruruima</subject><subject>Cytolytic activity</subject><subject>Cytotoxicity</subject><subject>Diaphragm - drug effects</subject><subject>Edema</subject><subject>Gel electrophoresis</subject><subject>High performance liquid chromatography</subject><subject>Isoelectric Point</subject><subject>Isoenzymes</subject><subject>Isoforms</subject><subject>Liquid chromatography</subject><subject>Mass spectrometry</subject><subject>Mass spectroscopy</subject><subject>Mice</subject><subject>Molecular Weight</subject><subject>Muscle, Skeletal - pathology</subject><subject>Muscles</subject><subject>Myoblasts</subject><subject>Myoblasts - drug effects</subject><subject>Myoblasts - metabolism</subject><subject>Myonecrosis</subject><subject>Myotubes</subject><subject>Necrosis - pathology</subject><subject>Neuromuscular junctions</subject><subject>Neuromuscular transmission</subject><subject>Phospholipase A2</subject><subject>Phospholipases A - chemistry</subject><subject>Phospholipases A - isolation & purification</subject><subject>Phospholipases A - toxicity</subject><subject>Phospholipases A2</subject><subject>Phrenic Nerve - drug effects</subject><subject>Proteins</subject><subject>Residues</subject><subject>Sequence Alignment</subject><subject>Skeletal muscle</subject><subject>Snakes</subject><subject>Sodium lauryl sulfate</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</subject><subject>Structural analysis</subject><subject>Structure-function relationships</subject><subject>Toxicity</subject><subject>Toxins</subject><subject>Venom</subject><issn>1572-3887</issn><issn>1875-8355</issn><issn>1573-4943</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNpFUctOwzAQtBCI8voALsjiTMCPJHaOEPGSKsEBztbWcSClics6BsFH8M24tBKnnR3vzNoeQo45O-eMqYvAWSVZxliZVSwXmdwie1yrItOyKLYTLlQitVYTsh_CnDGhKyV2yYQrwaSsyj3yc9V5--r6zsLijC5fAXuwfuFfVgSFoaFhxGjHiKm16Rjs6LD7hrHzA_UtHT89fZxeCtoF33rsA60bzLj40_5BSVv0Pa3Rj7CIgTYRuxASwIix64GGAd4c_XCD7w_JTguL4I429YA831w_1XfZ9OH2vr6cZlbkfFy9luu2Uc7NAGzRFK4EKSV3Epyo2EwWnCmlJci8aq12szzXiSlVo6EEXcoDcrr2XaJ_jy6MZu4jDmmlUYViKs9FlYb4esiiDwFda5aY7otfhjOzCsCsAzApALMKwMikOdkYx1nvmn_F5sflL2nkggM</recordid><startdate>200701</startdate><enddate>200701</enddate><creator>Ponce-Soto, 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pharmacological and structural characterization of two PLA2 isoforms Cdr-12 and Cdr-13 from Crotalus durissus ruruima snake venom</title><author>Ponce-Soto, Luis Alberto ; Baldasso, Paulo Aparecido ; Romero-Vargas, Frey Francisco ; Winck, Flávia V ; Novello, José Camillo ; Marangoni, Sergio</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c241t-90418fd7eebaac5d5e6a3331e3ae290b35107783a349fc8eb44810767d8a6a863</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Amino acid composition</topic><topic>Amino Acid Sequence</topic><topic>Amino acids</topic><topic>Animals</topic><topic>Biocompatibility</topic><topic>Cells, Cultured</topic><topic>Chemical bonds</topic><topic>Chemical Fractionation</topic><topic>Chickens</topic><topic>Crotalid Venoms - chemistry</topic><topic>Crotalid Venoms - enzymology</topic><topic>Crotalid Venoms - isolation & purification</topic><topic>Crotalid Venoms - toxicity</topic><topic>Crotalus</topic><topic>Crotalus durissus ruruima</topic><topic>Cytolytic activity</topic><topic>Cytotoxicity</topic><topic>Diaphragm - drug effects</topic><topic>Edema</topic><topic>Gel electrophoresis</topic><topic>High performance liquid chromatography</topic><topic>Isoelectric Point</topic><topic>Isoenzymes</topic><topic>Isoforms</topic><topic>Liquid chromatography</topic><topic>Mass spectrometry</topic><topic>Mass spectroscopy</topic><topic>Mice</topic><topic>Molecular Weight</topic><topic>Muscle, Skeletal - pathology</topic><topic>Muscles</topic><topic>Myoblasts</topic><topic>Myoblasts - drug effects</topic><topic>Myoblasts - metabolism</topic><topic>Myonecrosis</topic><topic>Myotubes</topic><topic>Necrosis - pathology</topic><topic>Neuromuscular junctions</topic><topic>Neuromuscular transmission</topic><topic>Phospholipase A2</topic><topic>Phospholipases A - chemistry</topic><topic>Phospholipases A - isolation & purification</topic><topic>Phospholipases A - toxicity</topic><topic>Phospholipases A2</topic><topic>Phrenic Nerve - drug effects</topic><topic>Proteins</topic><topic>Residues</topic><topic>Sequence Alignment</topic><topic>Skeletal muscle</topic><topic>Snakes</topic><topic>Sodium lauryl sulfate</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</topic><topic>Structural analysis</topic><topic>Structure-function relationships</topic><topic>Toxicity</topic><topic>Toxins</topic><topic>Venom</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ponce-Soto, Luis Alberto</creatorcontrib><creatorcontrib>Baldasso, Paulo Aparecido</creatorcontrib><creatorcontrib>Romero-Vargas, Frey Francisco</creatorcontrib><creatorcontrib>Winck, Flávia V</creatorcontrib><creatorcontrib>Novello, José Camillo</creatorcontrib><creatorcontrib>Marangoni, 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Edition</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><jtitle>The Protein Journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ponce-Soto, Luis Alberto</au><au>Baldasso, Paulo Aparecido</au><au>Romero-Vargas, Frey Francisco</au><au>Winck, Flávia V</au><au>Novello, José Camillo</au><au>Marangoni, Sergio</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Biochemical, pharmacological and structural characterization of two PLA2 isoforms Cdr-12 and Cdr-13 from Crotalus durissus ruruima snake venom</atitle><jtitle>The Protein Journal</jtitle><addtitle>Protein J</addtitle><date>2007-01</date><risdate>2007</risdate><volume>26</volume><issue>1</issue><spage>39</spage><epage>49</epage><pages>39-49</pages><issn>1572-3887</issn><eissn>1875-8355</eissn><eissn>1573-4943</eissn><abstract>Cdr-12 and Cdr-13 isoforms of PLA2, a D49 protein, were purified from Crotalus durissus ruruima venom after one chromatographic step, reverse phase HPLC on micro-Bondapack C-18. The molecular mass by SDS-PAGE of Cdr-12 and Cdr-13 isoforms of PLA2 was 14333.49 Da and 14296.42 Da, respectively and confirmed by MALDI-TOF mass spectrometry. The amino acid composition showed that both isoforms Cdr-12 and Cdr-13 have a high content of Lys, Tyr, Gly, Arg, and 14 half-Cys residues, typical of a basic PLA2. The isoforms Cdr-12 and Cdr-13 had a sequence of amino acids of 122 amino acid residues, being Cdr-12: SLLQFNKMIK FETRKNAIPF YAFYGCYCGW GGQGRPKDAT DRCCIVHDCC YGKLAKCNTK WDFYRYSLRS GYFQCGKGTW CEQQICECDR VAAECLRRSL STYRYGYMIY PDSRCREPSE TC and pI value 8.37 and Cdr-13: SLVQFEKMIK EETGKNAVPF YAFYGCYCGW GGRGRPKDAT DRCCIVHDCC YEKLVKCNTK WDFYRYSLRS GYFQCGKGTW CEQQICECDR VAAECLRRSL STYRYGKMIY PDSRCREPSE TC with a pI value of 8.13 This sequence shows high identity values when compared to other D49 PLA2s isolated from venoms of crotalics snakes. Skeletal muscle preparations from the young chicken have been previously used in order to study the effects of toxins on neuromuscular transmission, providing an important opportunity to study the differentiated behavior of a toxin before more than one model, because it shows differences in its sensibilities. In mice, the PLA2 isoforms Cdr-12 and Cdr-13 induced myonecrosis and edema, upon intramuscular or subcutaneous injections, respectively. In vitro, Cdr-12 and Cdr-13 isoforms of PLA2, caused a potent blockade of neuromuscular transmission in young chicken biventer cervicis preparation and produced cytotoxicity in murine C2C12 skeletal muscle myotubes and lack cytolytic activity upon myoblasts in vitro. Thus, the combined structural and functional information obtained identify Cdr-12 and Cdr-13 isoforms as members of the PLA2 family, which presents the typical bioactivities described for such proteins.</abstract><cop>Netherlands</cop><pub>Springer Nature B.V</pub><pmid>17203396</pmid><doi>10.1007/s10930-006-9042-3</doi><tpages>11</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1572-3887 |
| ispartof | The Protein Journal, 2007-01, Vol.26 (1), p.39-49 |
| issn | 1572-3887 1875-8355 1573-4943 |
| language | eng |
| recordid | cdi_proquest_journals_757074429 |
| source | MEDLINE; SpringerNature Journals |
| subjects | Amino acid composition Amino Acid Sequence Amino acids Animals Biocompatibility Cells, Cultured Chemical bonds Chemical Fractionation Chickens Crotalid Venoms - chemistry Crotalid Venoms - enzymology Crotalid Venoms - isolation & purification Crotalid Venoms - toxicity Crotalus Crotalus durissus ruruima Cytolytic activity Cytotoxicity Diaphragm - drug effects Edema Gel electrophoresis High performance liquid chromatography Isoelectric Point Isoenzymes Isoforms Liquid chromatography Mass spectrometry Mass spectroscopy Mice Molecular Weight Muscle, Skeletal - pathology Muscles Myoblasts Myoblasts - drug effects Myoblasts - metabolism Myonecrosis Myotubes Necrosis - pathology Neuromuscular junctions Neuromuscular transmission Phospholipase A2 Phospholipases A - chemistry Phospholipases A - isolation & purification Phospholipases A - toxicity Phospholipases A2 Phrenic Nerve - drug effects Proteins Residues Sequence Alignment Skeletal muscle Snakes Sodium lauryl sulfate Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Structural analysis Structure-function relationships Toxicity Toxins Venom |
| title | Biochemical, pharmacological and structural characterization of two PLA2 isoforms Cdr-12 and Cdr-13 from Crotalus durissus ruruima snake venom |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-22T12%3A53%3A22IST&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=Biochemical,%20pharmacological%20and%20structural%20characterization%20of%20two%20PLA2%20isoforms%20Cdr-12%20and%20Cdr-13%20from%20Crotalus%20durissus%20ruruima%20snake%20venom&rft.jtitle=The%20Protein%20Journal&rft.au=Ponce-Soto,%20Luis%20Alberto&rft.date=2007-01&rft.volume=26&rft.issue=1&rft.spage=39&rft.epage=49&rft.pages=39-49&rft.issn=1572-3887&rft.eissn=1875-8355&rft_id=info:doi/10.1007/s10930-006-9042-3&rft_dat=%3Cproquest_cross%3E2158060751%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=757074429&rft_id=info:pmid/17203396&rfr_iscdi=true |