Is rat an appropriate animal model to study the involvement of d‐serine catabolism in schizophrenia? insights from characterization of d‐amino acid oxidase
d‐Amino acid oxidase (DAAO; EC1.4.3.3) has been proposed to play a main role in the degradation of d‐serine, an allosteric activator of the N‐methyl‐d‐aspartate‐type glutamate receptor in the human brain, and to be associated with the onset of schizophrenia. To prevent excessive d‐serine degradation...
Gespeichert in:
| Veröffentlicht in: | The FEBS journal 2011-11, Vol.278 (22), p.4362-4373 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 4373 |
|---|---|
| container_issue | 22 |
| container_start_page | 4362 |
| container_title | The FEBS journal |
| container_volume | 278 |
| creator | Frattini, Luca F. Piubelli, Luciano Sacchi, Silvia Molla, Gianluca Pollegioni, Loredano |
| description | d‐Amino acid oxidase (DAAO; EC1.4.3.3) has been proposed to play a main role in the degradation of d‐serine, an allosteric activator of the N‐methyl‐d‐aspartate‐type glutamate receptor in the human brain, and to be associated with the onset of schizophrenia. To prevent excessive d‐serine degradation, novel drugs for schizophrenia treatment based on DAAO inhibition were designed and tested on rats. However, the properties of rat DAAO are unknown and various in vivo trials have demonstrated the effects of DAAO inhibitors on d‐serine concentration in rats. In the present study, rat DAAO was efficiently expressed in Escherichia coli. The recombinant enzyme was purified as an active, 40 kDa monomeric flavoenzyme showing the basic properties of the dehydrogenase‐oxidase class of flavoproteins. Rat DAAO differs significantly from the human counterpart because: (a) it possesses a different substrate specificity; (b) it shows a lower kinetic efficiency, mainly as a result of a low substrate affinity; (c) it differs in affinity for the binding of classical inhibitors; (d) it is a stable monomer in the absence of an active site ligand; and (e) it interacts with the mammalian protein modulator pLG72 yielding a ∼ 100 kDa complex in addition to the ∼ 200 kDa one, as formed by the human DAAO. Furthermore, the concentration of endogenous d‐serine in U87 glioblastoma cells was not affected by transfection with rat DAAO, whereas it was significantly decreased when expressing the human homologue. These results raise doubt on the use of the rat as a model system for testing new drugs against schizophrenia and indicate a different physiological function of DAAO in rodents and humans.
Structured digital
•
pLG72 binds rDAAO by molecular sieving (View interaction)
Novel drugs for schizophrenia treatment based on D‐amino acid oxidase (DAAO) inhibition were designed and tested on rats but the properties of rat DAAO are unknown. We demonstrated that rat DAAO differs from the human counterpart because it possesses a different substrate specificity; shows a lower substrate and ligand affinity; is a stable monomer and binds the modulator pLG72 yielding a different complex |
| doi_str_mv | 10.1111/j.1742-4658.2011.08354.x |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_901005566</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1221138255</sourcerecordid><originalsourceid>FETCH-LOGICAL-c5454-1036d3c27eddcf6d7d4f6d6d9082bbbed7d97f0d9da09dbf0c2613c5bf5f502e3</originalsourceid><addsrcrecordid>eNqNkcFuFCEYxydGY2v1FQzxopcdgRlmhoumNq02aeJBTbwRBr5x2AywAlN3e_IRfAPfzSeRcds9eDBygA_4ff8EfkWBCC5JHi_XJWlruqob1pUUE1LirmJ1ub1XHB8u7h_q-vNR8SjGNcYZ4vxhcUQJ7whu2-Pi52VEQSYkHZKbTfCbYGSCvDVWTsh6DRNKHsU06x1KIyDjrv10DRZcQn5A-tf3HxGCcYCUTLL3k4k2Qyiq0dz4zRjAGfk6n0TzZUwRDcFbpEYZpEq570Ym491dkrTGeSSV0chvjZYRHhcPBjlFeHK7nhSfLs4_nr1bXb1_e3l2erVSrGb1iuCq0ZWiLWithka3us5zoznuaN_3kA94O2DNtcRc9wNWtCGVYv3ABoYpVCfF831u_oOvM8QkrIkKpkk68HMUHBOMGWuaTL74J0koJaTqKGMZffYXuvZzcPkdS17b8aarM9TtIRV8jAEGkR1YGXaCYLHYFmuxiBSLVLHYFn9si21ufXqbP_cW9KHxTm8GXu2Bb2aC3X8Hi4vzNx-WsvoN2bO-Yw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>901789684</pqid></control><display><type>article</type><title>Is rat an appropriate animal model to study the involvement of d‐serine catabolism in schizophrenia? insights from characterization of d‐amino acid oxidase</title><source>Wiley Free Content</source><source>MEDLINE</source><source>IngentaConnect Free/Open Access Journals</source><source>Wiley Online Library Journals Frontfile Complete</source><source>Free Full-Text Journals in Chemistry</source><creator>Frattini, Luca F. ; Piubelli, Luciano ; Sacchi, Silvia ; Molla, Gianluca ; Pollegioni, Loredano</creator><creatorcontrib>Frattini, Luca F. ; Piubelli, Luciano ; Sacchi, Silvia ; Molla, Gianluca ; Pollegioni, Loredano</creatorcontrib><description>d‐Amino acid oxidase (DAAO; EC1.4.3.3) has been proposed to play a main role in the degradation of d‐serine, an allosteric activator of the N‐methyl‐d‐aspartate‐type glutamate receptor in the human brain, and to be associated with the onset of schizophrenia. To prevent excessive d‐serine degradation, novel drugs for schizophrenia treatment based on DAAO inhibition were designed and tested on rats. However, the properties of rat DAAO are unknown and various in vivo trials have demonstrated the effects of DAAO inhibitors on d‐serine concentration in rats. In the present study, rat DAAO was efficiently expressed in Escherichia coli. The recombinant enzyme was purified as an active, 40 kDa monomeric flavoenzyme showing the basic properties of the dehydrogenase‐oxidase class of flavoproteins. Rat DAAO differs significantly from the human counterpart because: (a) it possesses a different substrate specificity; (b) it shows a lower kinetic efficiency, mainly as a result of a low substrate affinity; (c) it differs in affinity for the binding of classical inhibitors; (d) it is a stable monomer in the absence of an active site ligand; and (e) it interacts with the mammalian protein modulator pLG72 yielding a ∼ 100 kDa complex in addition to the ∼ 200 kDa one, as formed by the human DAAO. Furthermore, the concentration of endogenous d‐serine in U87 glioblastoma cells was not affected by transfection with rat DAAO, whereas it was significantly decreased when expressing the human homologue. These results raise doubt on the use of the rat as a model system for testing new drugs against schizophrenia and indicate a different physiological function of DAAO in rodents and humans.
Structured digital
•
pLG72 binds rDAAO by molecular sieving (View interaction)
Novel drugs for schizophrenia treatment based on D‐amino acid oxidase (DAAO) inhibition were designed and tested on rats but the properties of rat DAAO are unknown. We demonstrated that rat DAAO differs from the human counterpart because it possesses a different substrate specificity; shows a lower substrate and ligand affinity; is a stable monomer and binds the modulator pLG72 yielding a different complex</description><identifier>ISSN: 1742-464X</identifier><identifier>EISSN: 1742-4658</identifier><identifier>DOI: 10.1111/j.1742-4658.2011.08354.x</identifier><identifier>PMID: 21981077</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Amino acids ; Animals ; Biochemistry ; Brain research ; Carrier Proteins - metabolism ; D-Amino-Acid Oxidase - genetics ; D-Amino-Acid Oxidase - metabolism ; Disease Models, Animal ; drug design ; d‐serine ; Enzyme Inhibitors - metabolism ; Flavin-Adenine Dinucleotide - metabolism ; flavoproteins ; Glioblastoma - metabolism ; Humans ; Inhibitor drugs ; ligand binding ; Protein Binding ; Protein Conformation ; Rats ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; Schizophrenia ; Serine - metabolism ; Substrate Specificity ; Tumor Cells, Cultured</subject><ispartof>The FEBS journal, 2011-11, Vol.278 (22), p.4362-4373</ispartof><rights>2011 The Authors Journal compilation © 2011 FEBS</rights><rights>2011 The Authors Journal compilation © 2011 FEBS.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5454-1036d3c27eddcf6d7d4f6d6d9082bbbed7d97f0d9da09dbf0c2613c5bf5f502e3</citedby><cites>FETCH-LOGICAL-c5454-1036d3c27eddcf6d7d4f6d6d9082bbbed7d97f0d9da09dbf0c2613c5bf5f502e3</cites></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.1742-4658.2011.08354.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1742-4658.2011.08354.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,1427,27901,27902,45550,45551,46384,46808</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21981077$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Frattini, Luca F.</creatorcontrib><creatorcontrib>Piubelli, Luciano</creatorcontrib><creatorcontrib>Sacchi, Silvia</creatorcontrib><creatorcontrib>Molla, Gianluca</creatorcontrib><creatorcontrib>Pollegioni, Loredano</creatorcontrib><title>Is rat an appropriate animal model to study the involvement of d‐serine catabolism in schizophrenia? insights from characterization of d‐amino acid oxidase</title><title>The FEBS journal</title><addtitle>FEBS J</addtitle><description>d‐Amino acid oxidase (DAAO; EC1.4.3.3) has been proposed to play a main role in the degradation of d‐serine, an allosteric activator of the N‐methyl‐d‐aspartate‐type glutamate receptor in the human brain, and to be associated with the onset of schizophrenia. To prevent excessive d‐serine degradation, novel drugs for schizophrenia treatment based on DAAO inhibition were designed and tested on rats. However, the properties of rat DAAO are unknown and various in vivo trials have demonstrated the effects of DAAO inhibitors on d‐serine concentration in rats. In the present study, rat DAAO was efficiently expressed in Escherichia coli. The recombinant enzyme was purified as an active, 40 kDa monomeric flavoenzyme showing the basic properties of the dehydrogenase‐oxidase class of flavoproteins. Rat DAAO differs significantly from the human counterpart because: (a) it possesses a different substrate specificity; (b) it shows a lower kinetic efficiency, mainly as a result of a low substrate affinity; (c) it differs in affinity for the binding of classical inhibitors; (d) it is a stable monomer in the absence of an active site ligand; and (e) it interacts with the mammalian protein modulator pLG72 yielding a ∼ 100 kDa complex in addition to the ∼ 200 kDa one, as formed by the human DAAO. Furthermore, the concentration of endogenous d‐serine in U87 glioblastoma cells was not affected by transfection with rat DAAO, whereas it was significantly decreased when expressing the human homologue. These results raise doubt on the use of the rat as a model system for testing new drugs against schizophrenia and indicate a different physiological function of DAAO in rodents and humans.
Structured digital
•
pLG72 binds rDAAO by molecular sieving (View interaction)
Novel drugs for schizophrenia treatment based on D‐amino acid oxidase (DAAO) inhibition were designed and tested on rats but the properties of rat DAAO are unknown. We demonstrated that rat DAAO differs from the human counterpart because it possesses a different substrate specificity; shows a lower substrate and ligand affinity; is a stable monomer and binds the modulator pLG72 yielding a different complex</description><subject>Amino acids</subject><subject>Animals</subject><subject>Biochemistry</subject><subject>Brain research</subject><subject>Carrier Proteins - metabolism</subject><subject>D-Amino-Acid Oxidase - genetics</subject><subject>D-Amino-Acid Oxidase - metabolism</subject><subject>Disease Models, Animal</subject><subject>drug design</subject><subject>d‐serine</subject><subject>Enzyme Inhibitors - metabolism</subject><subject>Flavin-Adenine Dinucleotide - metabolism</subject><subject>flavoproteins</subject><subject>Glioblastoma - metabolism</subject><subject>Humans</subject><subject>Inhibitor drugs</subject><subject>ligand binding</subject><subject>Protein Binding</subject><subject>Protein Conformation</subject><subject>Rats</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>Schizophrenia</subject><subject>Serine - metabolism</subject><subject>Substrate Specificity</subject><subject>Tumor Cells, Cultured</subject><issn>1742-464X</issn><issn>1742-4658</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkcFuFCEYxydGY2v1FQzxopcdgRlmhoumNq02aeJBTbwRBr5x2AywAlN3e_IRfAPfzSeRcds9eDBygA_4ff8EfkWBCC5JHi_XJWlruqob1pUUE1LirmJ1ub1XHB8u7h_q-vNR8SjGNcYZ4vxhcUQJ7whu2-Pi52VEQSYkHZKbTfCbYGSCvDVWTsh6DRNKHsU06x1KIyDjrv10DRZcQn5A-tf3HxGCcYCUTLL3k4k2Qyiq0dz4zRjAGfk6n0TzZUwRDcFbpEYZpEq570Ym491dkrTGeSSV0chvjZYRHhcPBjlFeHK7nhSfLs4_nr1bXb1_e3l2erVSrGb1iuCq0ZWiLWithka3us5zoznuaN_3kA94O2DNtcRc9wNWtCGVYv3ABoYpVCfF831u_oOvM8QkrIkKpkk68HMUHBOMGWuaTL74J0koJaTqKGMZffYXuvZzcPkdS17b8aarM9TtIRV8jAEGkR1YGXaCYLHYFmuxiBSLVLHYFn9si21ufXqbP_cW9KHxTm8GXu2Bb2aC3X8Hi4vzNx-WsvoN2bO-Yw</recordid><startdate>201111</startdate><enddate>201111</enddate><creator>Frattini, Luca F.</creator><creator>Piubelli, Luciano</creator><creator>Sacchi, Silvia</creator><creator>Molla, Gianluca</creator><creator>Pollegioni, Loredano</creator><general>Blackwell Publishing Ltd</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>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>201111</creationdate><title>Is rat an appropriate animal model to study the involvement of d‐serine catabolism in schizophrenia? insights from characterization of d‐amino acid oxidase</title><author>Frattini, Luca F. ; Piubelli, Luciano ; Sacchi, Silvia ; Molla, Gianluca ; Pollegioni, Loredano</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5454-1036d3c27eddcf6d7d4f6d6d9082bbbed7d97f0d9da09dbf0c2613c5bf5f502e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Amino acids</topic><topic>Animals</topic><topic>Biochemistry</topic><topic>Brain research</topic><topic>Carrier Proteins - metabolism</topic><topic>D-Amino-Acid Oxidase - genetics</topic><topic>D-Amino-Acid Oxidase - metabolism</topic><topic>Disease Models, Animal</topic><topic>drug design</topic><topic>d‐serine</topic><topic>Enzyme Inhibitors - metabolism</topic><topic>Flavin-Adenine Dinucleotide - metabolism</topic><topic>flavoproteins</topic><topic>Glioblastoma - metabolism</topic><topic>Humans</topic><topic>Inhibitor drugs</topic><topic>ligand binding</topic><topic>Protein Binding</topic><topic>Protein Conformation</topic><topic>Rats</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - metabolism</topic><topic>Schizophrenia</topic><topic>Serine - metabolism</topic><topic>Substrate Specificity</topic><topic>Tumor Cells, Cultured</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Frattini, Luca F.</creatorcontrib><creatorcontrib>Piubelli, Luciano</creatorcontrib><creatorcontrib>Sacchi, Silvia</creatorcontrib><creatorcontrib>Molla, Gianluca</creatorcontrib><creatorcontrib>Pollegioni, Loredano</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The FEBS journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Frattini, Luca F.</au><au>Piubelli, Luciano</au><au>Sacchi, Silvia</au><au>Molla, Gianluca</au><au>Pollegioni, Loredano</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Is rat an appropriate animal model to study the involvement of d‐serine catabolism in schizophrenia? insights from characterization of d‐amino acid oxidase</atitle><jtitle>The FEBS journal</jtitle><addtitle>FEBS J</addtitle><date>2011-11</date><risdate>2011</risdate><volume>278</volume><issue>22</issue><spage>4362</spage><epage>4373</epage><pages>4362-4373</pages><issn>1742-464X</issn><eissn>1742-4658</eissn><abstract>d‐Amino acid oxidase (DAAO; EC1.4.3.3) has been proposed to play a main role in the degradation of d‐serine, an allosteric activator of the N‐methyl‐d‐aspartate‐type glutamate receptor in the human brain, and to be associated with the onset of schizophrenia. To prevent excessive d‐serine degradation, novel drugs for schizophrenia treatment based on DAAO inhibition were designed and tested on rats. However, the properties of rat DAAO are unknown and various in vivo trials have demonstrated the effects of DAAO inhibitors on d‐serine concentration in rats. In the present study, rat DAAO was efficiently expressed in Escherichia coli. The recombinant enzyme was purified as an active, 40 kDa monomeric flavoenzyme showing the basic properties of the dehydrogenase‐oxidase class of flavoproteins. Rat DAAO differs significantly from the human counterpart because: (a) it possesses a different substrate specificity; (b) it shows a lower kinetic efficiency, mainly as a result of a low substrate affinity; (c) it differs in affinity for the binding of classical inhibitors; (d) it is a stable monomer in the absence of an active site ligand; and (e) it interacts with the mammalian protein modulator pLG72 yielding a ∼ 100 kDa complex in addition to the ∼ 200 kDa one, as formed by the human DAAO. Furthermore, the concentration of endogenous d‐serine in U87 glioblastoma cells was not affected by transfection with rat DAAO, whereas it was significantly decreased when expressing the human homologue. These results raise doubt on the use of the rat as a model system for testing new drugs against schizophrenia and indicate a different physiological function of DAAO in rodents and humans.
Structured digital
•
pLG72 binds rDAAO by molecular sieving (View interaction)
Novel drugs for schizophrenia treatment based on D‐amino acid oxidase (DAAO) inhibition were designed and tested on rats but the properties of rat DAAO are unknown. We demonstrated that rat DAAO differs from the human counterpart because it possesses a different substrate specificity; shows a lower substrate and ligand affinity; is a stable monomer and binds the modulator pLG72 yielding a different complex</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>21981077</pmid><doi>10.1111/j.1742-4658.2011.08354.x</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1742-464X |
| ispartof | The FEBS journal, 2011-11, Vol.278 (22), p.4362-4373 |
| issn | 1742-464X 1742-4658 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_901005566 |
| source | Wiley Free Content; MEDLINE; IngentaConnect Free/Open Access Journals; Wiley Online Library Journals Frontfile Complete; Free Full-Text Journals in Chemistry |
| subjects | Amino acids Animals Biochemistry Brain research Carrier Proteins - metabolism D-Amino-Acid Oxidase - genetics D-Amino-Acid Oxidase - metabolism Disease Models, Animal drug design d‐serine Enzyme Inhibitors - metabolism Flavin-Adenine Dinucleotide - metabolism flavoproteins Glioblastoma - metabolism Humans Inhibitor drugs ligand binding Protein Binding Protein Conformation Rats Recombinant Proteins - genetics Recombinant Proteins - metabolism Schizophrenia Serine - metabolism Substrate Specificity Tumor Cells, Cultured |
| title | Is rat an appropriate animal model to study the involvement of d‐serine catabolism in schizophrenia? insights from characterization of d‐amino acid oxidase |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-12T17%3A01%3A58IST&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=Is%20rat%20an%20appropriate%20animal%20model%20to%20study%20the%20involvement%20of%20d%E2%80%90serine%20catabolism%20in%20schizophrenia?%20insights%20from%20characterization%20of%20d%E2%80%90amino%20acid%20oxidase&rft.jtitle=The%20FEBS%20journal&rft.au=Frattini,%20Luca%20F.&rft.date=2011-11&rft.volume=278&rft.issue=22&rft.spage=4362&rft.epage=4373&rft.pages=4362-4373&rft.issn=1742-464X&rft.eissn=1742-4658&rft_id=info:doi/10.1111/j.1742-4658.2011.08354.x&rft_dat=%3Cproquest_cross%3E1221138255%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=901789684&rft_id=info:pmid/21981077&rfr_iscdi=true |