Plate ethanol-screening assay for selection of the Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability for xylose alcoholic fermentation

A new method for the selection of Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability to ferment xylose to ethanol was developed. The method is based on the ability of P. stipitis and H. polymorpha colonies to grow and produce ethanol on agar plates with xylose as the sole...

Full description

Saved in:
Bibliographic Details
Published in:Journal of industrial microbiology & biotechnology 2006-11, Vol.33 (11), p.934-940
Main Authors: Grabek-Lejko, D, Ryabova, O.B, Oklejewicz, B, Voronovsky, A.Y, Sibirny, A.A
Format: Article
Language:English
Subjects:
alcoholic fermentation
Bacteria
biofuels
Biological and medical sciences
Biotechnology
Culture Media
Energy sources
Enzymatic activity
enzyme activity
Ethanol
Ethanol - metabolism
ethanol production
Fermentation
Fundamental and applied biological sciences. Psychology
Hansenula polymorpha
lignocellulose
Microbiological Techniques
Microbiology
Mutagenesis, Insertional
Mutants
Mutation
Pichia - enzymology
Pichia - genetics
Pichia - growth & development
Pichia - metabolism
Pichia stipitis
Saccharomyces cerevisiae
xylose
Xylose - metabolism
Yeast
Yeasts
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-c412t-2d7e789f7664cc64f9486c0f0fa8117eeb2b2072eb40d80875d964895cd344433
cites cdi_FETCH-LOGICAL-c412t-2d7e789f7664cc64f9486c0f0fa8117eeb2b2072eb40d80875d964895cd344433
container_end_page 940
container_issue 11
container_start_page 934
container_title Journal of industrial microbiology & biotechnology
container_volume 33
creator Grabek-Lejko, D
Ryabova, O.B
Oklejewicz, B
Voronovsky, A.Y
Sibirny, A.A
description A new method for the selection of Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability to ferment xylose to ethanol was developed. The method is based on the ability of P. stipitis and H. polymorpha colonies to grow and produce ethanol on agar plates with xylose as the sole carbon and energy source. Secreted ethanol, in contrast to xylose, supports growth of cells of the indicator xylose-negative strains (the wild-type strain of Saccharomyces cerevisiae or Δxyl1 mutant of H. polymorpha) mixed with agar medium. The size of the tester culture-growth zone around xylose-grown colonies appeared to be dependent on the amount of secreted ethanol. Mutants with altered (decreased or elevated) ethanol production in xylose medium have been isolated using this method. The mutants exhibited pleiotropic alterations in enzymatic activities of the intermediary xylose metabolism.
doi_str_mv 10.1007/s10295-006-0147-7
format article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_754567811</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>754567811</sourcerecordid><originalsourceid>FETCH-LOGICAL-c412t-2d7e789f7664cc64f9486c0f0fa8117eeb2b2072eb40d80875d964895cd344433</originalsourceid><addsrcrecordid>eNpd0d1qFTEQB_BFFFurD-CNBkG8Wk2y-di9LEWtULCgvV7mZCfdlGyyJln0PI8vag7nQMGrCeQ3Mwn_pnnN6EdGqf6UGeWDbClVLWVCt_pJc16raqXs5NN67pRupejkWfMi5wdKqdSaP2_OmNJaql6dN39vPRQkWGYI0bfZJMTgwj2BnGFPbEwko0dTXAwkWlJmJLfOzA5ILm51xWUCYSLXEDKGzQNZo98vMa0zkD1CLmTZCoSSyW9XZgK-YMKJGFhh57wrxx1_9j5mrLcmztE7QyymBUOBw96XzTMLPuOrU71o7r58_nl13d58__rt6vKmNYLx0vJJo-4Hq5USxihhB9ErQy210DOmEXd8x6nmuBN06mmv5TQo0Q_STJ0Qousumg_HuWuKvzbMZVxcNug9BIxbHrUUUuk6q8p3_8mHuKVQHzcqxlWFWlbEjsikmHNCO67JLZD2I6PjIb_xmN9Y8xsP-Y269rw5Dd52C06PHafAKnh_ApANeJsgGJcfXX_4IT-4t0dnIY5wn6q5-8Ep6ygdWD_wofsHmtOvIw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>612675475</pqid></control><display><type>article</type><title>Plate ethanol-screening assay for selection of the Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability for xylose alcoholic fermentation</title><source>Open Access: Oxford University Press Open Journals</source><source>ABI/INFORM Global</source><creator>Grabek-Lejko, D ; Ryabova, O.B ; Oklejewicz, B ; Voronovsky, A.Y ; Sibirny, A.A</creator><creatorcontrib>Grabek-Lejko, D ; Ryabova, O.B ; Oklejewicz, B ; Voronovsky, A.Y ; Sibirny, A.A</creatorcontrib><description>A new method for the selection of Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability to ferment xylose to ethanol was developed. The method is based on the ability of P. stipitis and H. polymorpha colonies to grow and produce ethanol on agar plates with xylose as the sole carbon and energy source. Secreted ethanol, in contrast to xylose, supports growth of cells of the indicator xylose-negative strains (the wild-type strain of Saccharomyces cerevisiae or Δxyl1 mutant of H. polymorpha) mixed with agar medium. The size of the tester culture-growth zone around xylose-grown colonies appeared to be dependent on the amount of secreted ethanol. Mutants with altered (decreased or elevated) ethanol production in xylose medium have been isolated using this method. The mutants exhibited pleiotropic alterations in enzymatic activities of the intermediary xylose metabolism.</description><identifier>ISSN: 1367-5435</identifier><identifier>EISSN: 1476-5535</identifier><identifier>DOI: 10.1007/s10295-006-0147-7</identifier><identifier>PMID: 16775686</identifier><language>eng</language><publisher>Heidelberg: Springer</publisher><subject>alcoholic fermentation ; Bacteria ; biofuels ; Biological and medical sciences ; Biotechnology ; Culture Media ; Energy sources ; Enzymatic activity ; enzyme activity ; Ethanol ; Ethanol - metabolism ; ethanol production ; Fermentation ; Fundamental and applied biological sciences. Psychology ; Hansenula polymorpha ; lignocellulose ; Microbiological Techniques ; Microbiology ; Mutagenesis, Insertional ; Mutants ; Mutation ; Pichia - enzymology ; Pichia - genetics ; Pichia - growth &amp; development ; Pichia - metabolism ; Pichia stipitis ; Saccharomyces cerevisiae ; xylose ; Xylose - metabolism ; Yeast ; Yeasts</subject><ispartof>Journal of industrial microbiology &amp; biotechnology, 2006-11, Vol.33 (11), p.934-940</ispartof><rights>2006 INIST-CNRS</rights><rights>Society for Industrial Microbiology 2006</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c412t-2d7e789f7664cc64f9486c0f0fa8117eeb2b2072eb40d80875d964895cd344433</citedby><cites>FETCH-LOGICAL-c412t-2d7e789f7664cc64f9486c0f0fa8117eeb2b2072eb40d80875d964895cd344433</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/612675475/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$H</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/612675475?pq-origsite=primo$$EHTML$$P50$$Gproquest$$H</linktohtml><link.rule.ids>314,780,784,11688,27924,27925,36060,36061,44363,74895</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=18207226$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16775686$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Grabek-Lejko, D</creatorcontrib><creatorcontrib>Ryabova, O.B</creatorcontrib><creatorcontrib>Oklejewicz, B</creatorcontrib><creatorcontrib>Voronovsky, A.Y</creatorcontrib><creatorcontrib>Sibirny, A.A</creatorcontrib><title>Plate ethanol-screening assay for selection of the Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability for xylose alcoholic fermentation</title><title>Journal of industrial microbiology &amp; biotechnology</title><addtitle>J Ind Microbiol Biotechnol</addtitle><description>A new method for the selection of Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability to ferment xylose to ethanol was developed. The method is based on the ability of P. stipitis and H. polymorpha colonies to grow and produce ethanol on agar plates with xylose as the sole carbon and energy source. Secreted ethanol, in contrast to xylose, supports growth of cells of the indicator xylose-negative strains (the wild-type strain of Saccharomyces cerevisiae or Δxyl1 mutant of H. polymorpha) mixed with agar medium. The size of the tester culture-growth zone around xylose-grown colonies appeared to be dependent on the amount of secreted ethanol. Mutants with altered (decreased or elevated) ethanol production in xylose medium have been isolated using this method. The mutants exhibited pleiotropic alterations in enzymatic activities of the intermediary xylose metabolism.</description><subject>alcoholic fermentation</subject><subject>Bacteria</subject><subject>biofuels</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Culture Media</subject><subject>Energy sources</subject><subject>Enzymatic activity</subject><subject>enzyme activity</subject><subject>Ethanol</subject><subject>Ethanol - metabolism</subject><subject>ethanol production</subject><subject>Fermentation</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hansenula polymorpha</subject><subject>lignocellulose</subject><subject>Microbiological Techniques</subject><subject>Microbiology</subject><subject>Mutagenesis, Insertional</subject><subject>Mutants</subject><subject>Mutation</subject><subject>Pichia - enzymology</subject><subject>Pichia - genetics</subject><subject>Pichia - growth &amp; development</subject><subject>Pichia - metabolism</subject><subject>Pichia stipitis</subject><subject>Saccharomyces cerevisiae</subject><subject>xylose</subject><subject>Xylose - metabolism</subject><subject>Yeast</subject><subject>Yeasts</subject><issn>1367-5435</issn><issn>1476-5535</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>M0C</sourceid><recordid>eNpd0d1qFTEQB_BFFFurD-CNBkG8Wk2y-di9LEWtULCgvV7mZCfdlGyyJln0PI8vag7nQMGrCeQ3Mwn_pnnN6EdGqf6UGeWDbClVLWVCt_pJc16raqXs5NN67pRupejkWfMi5wdKqdSaP2_OmNJaql6dN39vPRQkWGYI0bfZJMTgwj2BnGFPbEwko0dTXAwkWlJmJLfOzA5ILm51xWUCYSLXEDKGzQNZo98vMa0zkD1CLmTZCoSSyW9XZgK-YMKJGFhh57wrxx1_9j5mrLcmztE7QyymBUOBw96XzTMLPuOrU71o7r58_nl13d58__rt6vKmNYLx0vJJo-4Hq5USxihhB9ErQy210DOmEXd8x6nmuBN06mmv5TQo0Q_STJ0Qousumg_HuWuKvzbMZVxcNug9BIxbHrUUUuk6q8p3_8mHuKVQHzcqxlWFWlbEjsikmHNCO67JLZD2I6PjIb_xmN9Y8xsP-Y269rw5Dd52C06PHafAKnh_ApANeJsgGJcfXX_4IT-4t0dnIY5wn6q5-8Ep6ygdWD_wofsHmtOvIw</recordid><startdate>20061101</startdate><enddate>20061101</enddate><creator>Grabek-Lejko, D</creator><creator>Ryabova, O.B</creator><creator>Oklejewicz, B</creator><creator>Voronovsky, A.Y</creator><creator>Sibirny, A.A</creator><general>Springer</general><general>Oxford University Press</general><scope>FBQ</scope><scope>IQODW</scope><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>3V.</scope><scope>7QL</scope><scope>7QR</scope><scope>7T7</scope><scope>7WY</scope><scope>7WZ</scope><scope>7X7</scope><scope>7XB</scope><scope>87Z</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8FL</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BEZIV</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FRNLG</scope><scope>FYUFA</scope><scope>F~G</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>HCIFZ</scope><scope>K60</scope><scope>K6~</scope><scope>K9.</scope><scope>L.-</scope><scope>LK8</scope><scope>M0C</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>P64</scope><scope>PQBIZ</scope><scope>PQBZA</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7QO</scope></search><sort><creationdate>20061101</creationdate><title>Plate ethanol-screening assay for selection of the Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability for xylose alcoholic fermentation</title><author>Grabek-Lejko, D ; Ryabova, O.B ; Oklejewicz, B ; Voronovsky, A.Y ; Sibirny, A.A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c412t-2d7e789f7664cc64f9486c0f0fa8117eeb2b2072eb40d80875d964895cd344433</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>alcoholic fermentation</topic><topic>Bacteria</topic><topic>biofuels</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Culture Media</topic><topic>Energy sources</topic><topic>Enzymatic activity</topic><topic>enzyme activity</topic><topic>Ethanol</topic><topic>Ethanol - metabolism</topic><topic>ethanol production</topic><topic>Fermentation</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hansenula polymorpha</topic><topic>lignocellulose</topic><topic>Microbiological Techniques</topic><topic>Microbiology</topic><topic>Mutagenesis, Insertional</topic><topic>Mutants</topic><topic>Mutation</topic><topic>Pichia - enzymology</topic><topic>Pichia - genetics</topic><topic>Pichia - growth &amp; development</topic><topic>Pichia - metabolism</topic><topic>Pichia stipitis</topic><topic>Saccharomyces cerevisiae</topic><topic>xylose</topic><topic>Xylose - metabolism</topic><topic>Yeast</topic><topic>Yeasts</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Grabek-Lejko, D</creatorcontrib><creatorcontrib>Ryabova, O.B</creatorcontrib><creatorcontrib>Oklejewicz, B</creatorcontrib><creatorcontrib>Voronovsky, A.Y</creatorcontrib><creatorcontrib>Sibirny, A.A</creatorcontrib><collection>AGRIS</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>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Chemoreception Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>ABI/INFORM Collection</collection><collection>ABI/INFORM Global (PDF only)</collection><collection>Health &amp; Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ABI/INFORM Collection</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</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>ABI/INFORM Collection (Alumni Edition)</collection><collection>Research Library (Alumni Edition)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>Business Premium Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central</collection><collection>Engineering Research Database</collection><collection>Business Premium Collection (Alumni)</collection><collection>Health Research Premium Collection</collection><collection>ABI/INFORM Global (Corporate)</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>Research Library Prep</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Business Collection (Alumni Edition)</collection><collection>ProQuest Business Collection</collection><collection>ProQuest Health &amp; Medical Complete (Alumni)</collection><collection>ABI/INFORM Professional Advanced</collection><collection>ProQuest Biological Science Collection</collection><collection>ABI/INFORM Global</collection><collection>Health &amp; Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>ProQuest research library</collection><collection>ProQuest Science Journals</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Research Library (Corporate)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>One Business</collection><collection>ProQuest One Business (Alumni)</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 Basic</collection><collection>Biotechnology Research Abstracts</collection><jtitle>Journal of industrial microbiology &amp; biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Grabek-Lejko, D</au><au>Ryabova, O.B</au><au>Oklejewicz, B</au><au>Voronovsky, A.Y</au><au>Sibirny, A.A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Plate ethanol-screening assay for selection of the Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability for xylose alcoholic fermentation</atitle><jtitle>Journal of industrial microbiology &amp; biotechnology</jtitle><addtitle>J Ind Microbiol Biotechnol</addtitle><date>2006-11-01</date><risdate>2006</risdate><volume>33</volume><issue>11</issue><spage>934</spage><epage>940</epage><pages>934-940</pages><issn>1367-5435</issn><eissn>1476-5535</eissn><abstract>A new method for the selection of Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability to ferment xylose to ethanol was developed. The method is based on the ability of P. stipitis and H. polymorpha colonies to grow and produce ethanol on agar plates with xylose as the sole carbon and energy source. Secreted ethanol, in contrast to xylose, supports growth of cells of the indicator xylose-negative strains (the wild-type strain of Saccharomyces cerevisiae or Δxyl1 mutant of H. polymorpha) mixed with agar medium. The size of the tester culture-growth zone around xylose-grown colonies appeared to be dependent on the amount of secreted ethanol. Mutants with altered (decreased or elevated) ethanol production in xylose medium have been isolated using this method. The mutants exhibited pleiotropic alterations in enzymatic activities of the intermediary xylose metabolism.</abstract><cop>Heidelberg</cop><pub>Springer</pub><pmid>16775686</pmid><doi>10.1007/s10295-006-0147-7</doi><tpages>7</tpages></addata></record>
fulltext fulltext
identifier ISSN: 1367-5435
ispartof Journal of industrial microbiology & biotechnology, 2006-11, Vol.33 (11), p.934-940
issn 1367-5435
1476-5535
language eng
recordid cdi_proquest_miscellaneous_754567811
source Open Access: Oxford University Press Open Journals; ABI/INFORM Global
subjects alcoholic fermentation
Bacteria
biofuels
Biological and medical sciences
Biotechnology
Culture Media
Energy sources
Enzymatic activity
enzyme activity
Ethanol
Ethanol - metabolism
ethanol production
Fermentation
Fundamental and applied biological sciences. Psychology
Hansenula polymorpha
lignocellulose
Microbiological Techniques
Microbiology
Mutagenesis, Insertional
Mutants
Mutation
Pichia - enzymology
Pichia - genetics
Pichia - growth & development
Pichia - metabolism
Pichia stipitis
Saccharomyces cerevisiae
xylose
Xylose - metabolism
Yeast
Yeasts
title Plate ethanol-screening assay for selection of the Pichia stipitis and Hansenula polymorpha yeast mutants with altered capability for xylose alcoholic fermentation
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-04T23%3A53%3A29IST&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=Plate%20ethanol-screening%20assay%20for%20selection%20of%20the%20Pichia%20stipitis%20and%20Hansenula%20polymorpha%20yeast%20mutants%20with%20altered%20capability%20for%20xylose%20alcoholic%20fermentation&rft.jtitle=Journal%20of%20industrial%20microbiology%20&%20biotechnology&rft.au=Grabek-Lejko,%20D&rft.date=2006-11-01&rft.volume=33&rft.issue=11&rft.spage=934&rft.epage=940&rft.pages=934-940&rft.issn=1367-5435&rft.eissn=1476-5535&rft_id=info:doi/10.1007/s10295-006-0147-7&rft_dat=%3Cproquest_cross%3E754567811%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c412t-2d7e789f7664cc64f9486c0f0fa8117eeb2b2072eb40d80875d964895cd344433%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=612675475&rft_id=info:pmid/16775686&rfr_iscdi=true