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Subject: ASPM-associated stem cell proliferation is involved in malignant progression of gliomas and constitutes an attractive therapeutic target
Date: Sat, 3 Apr 2010 18:53:51 +0200
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A>=20
      =C2=A92010 Bikeye et al; licensee BioMed Central Ltd.</DIV>
      <DIV class=3Dfm-title>ASPM-associated stem cell proliferation is =
involved in=20
      malignant progression of gliomas and constitutes an attractive =
therapeutic=20
      target</DIV>
      <DIV class=3D"contrib-group fm-author">Sandra-Nadia Ngwabyt=20
      Bikeye,<SUP>1</SUP> Carole Colin,<SUP>2</SUP> Yannick =
Marie,<SUP>1</SUP>=20
      Rapha=C3=ABl Vampouille,<SUP>4</SUP> Philippe =
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      Idbaih,<SUP>1,</SUP><SUP>3</SUP> Charles F=C3=A9lix =
Calvo,<SUP>1</SUP> Pascal=20
      Leuraud,<SUP>6</SUP> Myriam Lassalle,<SUP>6</SUP> Soufiane El=20
      Hallani,<SUP>1</SUP> Jean-Yves Delattre,<SUP>1,</SUP><SUP>3</SUP> =
and Marc=20
      Sanson<SUP><IMG alt=3D"corresponding author"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/corehtml/pmc/pmcgifs/corrauth.gif"></S=
UP><SUP>1,</SUP><SUP>3</SUP></DIV>
      <DIV class=3Dfm-affl><SUP>1</SUP>UMR975 INSERM-UPMC, GH =
Piti=C3=A9- Salp=C3=AAtri=C3=A8re,=20
      Paris</DIV>
      <DIV class=3Dfm-affl><SUP>2</SUP>UMR911-CRO2, Facult=C3=A9 de =
M=C3=A9decine Timone,=20
      Universit=C3=A9 de la M=C3=A9diterran=C3=A9e, Marseille</DIV>
      <DIV class=3Dfm-affl><SUP>3</SUP>Service de Neurologie Mazarin, GH =
Piti=C3=A9-=20
      Salp=C3=AAtri=C3=A8re, APHP, Paris</DIV>
      <DIV class=3Dfm-affl><SUP>4</SUP>Biotechnology &amp; Biotherapy =
laboratory,=20
      Centre de Recherche de l'Institut du Cerveau et de la Moelle, CNRS =

      UMR9225, INSERM UMRS 975 Universit=C3=A9 Pierre et Marie Curie, =
H=C3=B4pital de la=20
      Salp=C3=AAtri=C3=A8re, Paris</DIV>
      <DIV class=3Dfm-affl><SUP>5</SUP>Service de Neuropathologie =
Raymond=20
      Escourolle, GH Piti=C3=A9- Salp=C3=AAtri=C3=A8re, Paris</DIV>
      <DIV class=3Dfm-affl><SUP>6</SUP>XenTech, Evry, France</DIV>
      <DIV class=3Dfm-footnote><SUP><IMG alt=3D"corresponding author"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/corehtml/pmc/pmcgifs/corrauth.gif"></S=
UP>Corresponding=20
      author.</DIV>
      <DIV class=3Dfm-footnote><SPAN class=3Dcontrib-email =
id=3DA1>Sandra-Nadia=20
      Ngwabyt Bikeye: <SPAN class=3De_id703161>nmjfr/at/yahoo.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id703161", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.oohay/ta/rfjmn:otliam', '/at/', '@') + '">' =
+ reverseAndReplaceString('rf.oohay/ta/rfjmn', '/at/','@') + =
'</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA2>Carole Colin: <SPAN=20
      class=3De_id429433>carole.colin/at/univmed.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id429433", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.demvinu/ta/niloc.elorac:otliam', '/at/', =
'@') + '">' + reverseAndReplaceString('rf.demvinu/ta/niloc.elorac', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA3>Yannick Marie: <SPAN=20
      class=3De_id429459>yannick.marie/at/upmc.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id429459", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.cmpu/ta/eiram.kcinnay:otliam', '/at/', '@') =
+ '">' + reverseAndReplaceString('rf.cmpu/ta/eiram.kcinnay', '/at/','@') =
+ '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA4>Rapha=C3=ABl =
Vampouille: <SPAN=20
      class=3De_id139749>raphael.vampouille/at/upmc.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id139749", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.cmpu/ta/elliuopmav.leahpar:otliam', '/at/', =
'@') + '">' + reverseAndReplaceString('rf.cmpu/ta/elliuopmav.leahpar', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA5>Philippe Ravassard: =
<SPAN=20
      class=3De_id139774>ravassar/at/chups.jussieu.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id139774", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.ueissuj.spuhc/ta/rassavar:otliam', '/at/', =
'@') + '">' + reverseAndReplaceString('rf.ueissuj.spuhc/ta/rassavar', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA6>Audrey Rousseau: =
<SPAN=20
      class=3De_id139801>audrey.rousseau/at/psl.aphp.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id139801", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.phpa.lsp/ta/uaessuor.yerdua:otliam', '/at/', =
'@') + '">' + reverseAndReplaceString('rf.phpa.lsp/ta/uaessuor.yerdua', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA7>Blandine Boisselier: =
<SPAN=20
      class=3De_id705519>boisselierblandine/at/yahoo.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id705519", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.oohay/ta/enidnalbreilessiob:otliam', '/at/', =
'@') + '">' + reverseAndReplaceString('rf.oohay/ta/enidnalbreilessiob', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA8>Ahmed Idbaih: <SPAN=20
      class=3De_id705554>ahmed.idbaih/at/gmail.com</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id705554", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('moc.liamg/ta/hiabdi.demha:otliam', '/at/', '@') =
+ '">' + reverseAndReplaceString('moc.liamg/ta/hiabdi.demha', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA9>Charles F=C3=A9lix =
Calvo: <SPAN=20
      =
class=3De_id705579>charles-felix.calvo/at/college-de-france.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id705579", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.ecnarf\-ed\-egelloc/ta/ovlac.xilef\-selrahc:o=
tliam', '/at/', '@') + '">' + =
reverseAndReplaceString('rf.ecnarf\-ed\-egelloc/ta/ovlac.xilef\-selrahc',=
 '/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA10>Pascal Leuraud: =
<SPAN=20
      class=3De_id705606>Pascal.leuraud/at/hotmail.com</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id705606", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('moc.liamtoh/ta/duaruel.lacsaP:otliam', '/at/', =
'@') + '">' + reverseAndReplaceString('moc.liamtoh/ta/duaruel.lacsaP', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA11>Myriam Lassalle: =
<SPAN=20
      class=3De_id783213>myriam.lassalle/at/hotmail.com</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id783213", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('moc.liamtoh/ta/ellassal.mairym:otliam', '/at/', =
'@') + '">' + reverseAndReplaceString('moc.liamtoh/ta/ellassal.mairym', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA12>Soufiane El Hallani: =
<SPAN=20
      class=3De_id783238>elhallanisoufiane/at/yahoo.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id783238", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.oohay/ta/enaifuosinallahle:otliam', '/at/', =
'@') + '">' + reverseAndReplaceString('rf.oohay/ta/enaifuosinallahle', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA13>Jean-Yves Delattre: =
<SPAN=20
      class=3De_id783273>jean-yves.delattre/at/psl.aphp.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id783273", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.phpa.lsp/ta/erttaled.sevy\-naej:otliam', =
'/at/', '@') + '">' + =
reverseAndReplaceString('rf.phpa.lsp/ta/erttaled.sevy\-naej', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      ; </SPAN><SPAN class=3Dcontrib-email id=3DA14>Marc Sanson: <SPAN=20
      class=3De_id783310>marc.sanson/at/psl.aphp.fr</SPAN>
      <SCRIPT language=3DJavaScript type=3Dtext/javascript><!--=0A=
                                    try{initUnObscureEmail =
("e_id783310", '<a class=3D"ext-reflink" href=3D"' + =
reverseAndReplaceString('rf.phpa.lsp/ta/nosnas.cram:otliam', '/at/', =
'@') + '">' + reverseAndReplaceString('rf.phpa.lsp/ta/nosnas.cram', =
'/at/','@') + '</a>')}catch(e){}=0A=
                                //--></SCRIPT>
      </SPAN> </DIV>
      <DIV class=3Dfm-pubdate>Received September 13, 2009; Accepted =
January 11,=20
      2010.</DIV>
      <DIV class=3Dfm-copyright>This is an Open Access article =
distributed under=20
      the terms of the Creative Commons Attribution License (<SPAN=20
      class=3Dext-reflink><A class=3Dext-reflink =
onclick=3D"focuswin('pmc_ext')"=20
      href=3D"http://creativecommons.org/licenses/by/2.0" =
target=3Dpmc_ext=20
      =
ref=3D"reftype=3Dextlink&amp;article-id=3D2817685&amp;issue-id=3D184705&a=
mp;journal-id=3D96&amp;FROM=3DArticle%7CFront%20Matter&amp;TO=3DExternal%=
7CLink%7CURI&amp;rendering-type=3Dnormal">http://creativecommons.org/lice=
nses/by/2.0</A></SPAN>),=20
      which permits unrestricted use, distribution, and reproduction in =
any=20
      medium, provided the original work is properly cited.</DIV></DIV>
      <DIV class=3Dsec id=3D__abstractid683998>
      <DIV class=3D"head1 section-title" =
id=3D__abstractid683998titletitle=20
      style=3D"TEXT-TRANSFORM: none">
      <DIV class=3Dother-sections>
      <UL class=3D"noext-menu noext-menu-hidden">
        <LI><A class=3Dfirst-link=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">&nbsp;Other=20
        Sections=E2=96=BC</A>
        <UL class=3D"submenu head1">
          <LI class=3D"submenu-item current-item"><A class=3D""=20
          style=3D"TEXT-TRANSFORM: none" onclick=3Dreturn(false)=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">Abstract</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid684043">Introduction</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid834380">Materials=20
          and methods</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid828904">Results</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid704257">Discussion</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839339">Competing=20
          interests</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839346">Authors'=20
          contributions</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839355">Supplementary=20
          Material</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__ref-listid731707">References</A></LI></UL></LI></UL></DIV>
      <DIV>Abstract</DIV></DIV>
      <DIV class=3Dsection-content =
id=3D__abstractid683998content><!--article-meta-->
      <DIV class=3D"sec sec-first" id=3D__secid683999>
      <DIV class=3D"abs-head2 head-separate">Background</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid684003>ASPM (<EM>Abnormal =
Spindle-like=20
      Microcephaly associated</EM>) over-expression was recently =
implicated in=20
      the development of malignant gliomas.</DIV></DIV>
      <DIV class=3Dsec id=3D__secid684010>
      <DIV class=3D"abs-head2 head-separate">Results</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid684014>To better =
characterize the=20
      involvement of ASPM in gliomas, we investigated the mRNA =
expression in 175=20
      samples, including 8 WHO Grade II, 75 WHO Grade III and 92 WHO =
Grade IV=20
      tumors. <EM>Aspm </EM>expression was strongly correlated with =
tumor grade=20
      and increased at recurrence when compared to the initial lesion, =
whatever=20
      the initial grade of the primary tumor. ASPM expression also =
increased=20
      over serial passages in gliomaspheres <EM>in vitro </EM>and in =
mouse=20
      xenografts <EM>in vivo</EM>. Lentivirus-mediated shRNA silencing =
of ASPM=20
      resulted in dramatic proliferation arrest and cell death in two =
different=20
      gliomasphere models.</DIV></DIV>
      <DIV class=3D"sec sec-last" id=3D__secid684033>
      <DIV class=3D"abs-head2 head-separate">Conclusion</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid684037>These data suggest =
that ASPM is=20
      involved in the malignant progression of gliomas, possibly through =

      expansion of a cancer stem cell compartment, and is an attractive=20
      therapeutic target in glioblastoma =
multiforme.</DIV></DIV></DIV></DIV>
      <DIV class=3Dsec id=3D__secid684043>
      <DIV class=3D"head1 section-title" id=3D__secid684043titletitle=20
      style=3D"TEXT-TRANSFORM: none">
      <DIV class=3Dother-sections>
      <UL class=3D"noext-menu noext-menu-hidden">
        <LI><A class=3Dfirst-link=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">&nbsp;Other=20
        Sections=E2=96=BC</A>
        <UL class=3D"submenu head1">
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__abstractid683998">Abstract</A>
          <LI class=3D"submenu-item current-item"><A class=3D""=20
          style=3D"TEXT-TRANSFORM: none" onclick=3Dreturn(false)=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">Introduction</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid834380">Materials=20
          and methods</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid828904">Results</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid704257">Discussion</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839339">Competing=20
          interests</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839346">Authors'=20
          contributions</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839355">Supplementary=20
          Material</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__ref-listid731707">References</A></LI></UL></LI></UL></DIV>
      <DIV>Introduction</DIV></DIV>
      <DIV class=3Dsection-content id=3D__secid684043content>
      <DIV class=3D"p p-first" id=3D__pid684047>Gliomas are the most =
common type of=20
      primary tumors in the central nervous system (CNS), and their =
usual fate=20
      is to recur, either as a similar or more frequently as a higher =
grade=20
      lesion, explaining their grim prognosis. The cellular origin of =
gliomas=20
      remains unknown, but accumulating data suggest that transformed, =
stem-like=20
      precursors (CSC) may represent the cells that drive their growth =
[<A=20
      class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/15466194"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B1">1</A>,<A class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/15549107"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B2">2</A>].</DIV>
      <DIV class=3Dp id=3D__pid684075>The product <EM>of ASPM </EM>gene=20
      (<EM>Abnormal Spindle-like Microcephaly associated</EM>) localizes =
to=20
      centrosomes, spindle poles, and the midbody [<A=20
      class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/16798874"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B3">3</A>]. By promoting neuroblast proliferation and =
driving the=20
      orientation of mitotic cleavage, it allows for symmetric, =
proliferative=20
      division of neuroepithelial cells during brain development, and=20
      consequently brain surface expansion [<A class=3D"cite-reflink =
bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/16798874"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B3">3</A>]. Mutations within this gene produce primary =
microcephaly=20
      [<A class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/12355089"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B4">4</A>]. In addition to its role in embryonic =
development, ASPM is=20
      highly expressed in many tumor cell lines [<A=20
      class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/15972725"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B5">5</A>], suggesting an important involvement of this =
gene during=20
      tumorigenesis. Recent evidence suggests that ASPM is also =
overexpressed in=20
      malignant gliomas and that its knockdown inhibits tumor =
proliferation [<A=20
      class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/17090670"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B6">6</A>,<A class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18636190"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B7">7</A>].</DIV>
      <DIV class=3D"p p-last" id=3D__pid834367>In this study, we report =
that ASPM=20
      expression is correlated with tumor grade and increases at =
recurrence or=20
      after consecutive passages <EM>in vitro </EM>and <EM>in vivo</EM>. =

      Furthermore, we found that shRNA-mediated inhibition of ASPM =
results in=20
      dramatic growth inhibition and extensive cell death in=20
      gliomaspheres.</DIV></DIV></DIV>
      <DIV class=3Dsec id=3D__secid834380>
      <DIV class=3D"head1 section-title" id=3D__secid834380titletitle=20
      style=3D"TEXT-TRANSFORM: none">
      <DIV class=3Dother-sections>
      <UL class=3D"noext-menu noext-menu-hidden">
        <LI><A class=3Dfirst-link=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">&nbsp;Other=20
        Sections=E2=96=BC</A>
        <UL class=3D"submenu head1">
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__abstractid683998">Abstract</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid684043">Introduction</A>
          <LI class=3D"submenu-item current-item"><A class=3D""=20
          style=3D"TEXT-TRANSFORM: none" onclick=3Dreturn(false)=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">Materials=20
          and methods</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid828904">Results</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid704257">Discussion</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839339">Competing=20
          interests</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839346">Authors'=20
          contributions</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839355">Supplementary=20
          Material</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__ref-listid731707">References</A></LI></UL></LI></UL></DIV>
      <DIV>Materials and methods</DIV></DIV>
      <DIV class=3Dsection-content id=3D__secid834380content>
      <DIV class=3D"sec sec-first" id=3D__secid834386>
      <DIV class=3D"head2 head-separate">Patients and tissue =
samples</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid834390>Tumor samples =
frozen in liquid=20
      nitrogen were obtained from patients who underwent surgery for a =
glioma at=20
      our institution and gave informed consent for molecular research =
studies.=20
      Histological diagnoses were made according to the World Health=20
      Organization (WHO). BAC-array-based Comparative Genomic =
Hybridization=20
      analysis (CGH-array) was performed as previously described [<A=20
      class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18076069"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B8">8</A>]. Non-tumoral (NT) control brain (cerebellum and =
white=20
      matter) was obtained from injured or epileptic patients =
(cryptogenetic=20
      epilepsy) who underwent surgery.</DIV></DIV>
      <DIV class=3Dsec id=3D__secid452819>
      <DIV class=3D"head2 head-separate">Total RNA isolation, reverse=20
      transcription (RT) and quantitative real-time PCR (qPCR)</DIV>
      <DIV class=3D"p p-first" id=3D__pid452823>Total RNA extraction was =
performed=20
      using RNAble blue solution (Eurobio, France) according to the=20
      manufacturer's instructions. Total RNA was systematically analyzed =
on both=20
      a spectrophotometer and an Agilent 2100 bioanalyzer (Agilent =
Technologies,=20
      FRANCE). cDNA was prepared from 1 =CE=BCg RNA using a combination =
of random=20
      primers (48190-011; Invitrogen, France) and MMLV reverse =
transcriptase=20
      (28025-013; Invitrogen, France).</DIV>
      <DIV class=3D"p p-last" id=3D__pid452829>Gene expression was =
quantified by=20
      Syber Green real-time qPCR analysis (Absolute solute Syber Green =
Rox Mix,=20
      Abgene, Paris, France). Real-time qPCR cycles were as follows: =
initial=20
      denaturation at 95=C2=B0C for 15 minutes, 95=C2=B0C for 30 seconds =
and annealing at=20
      60=C2=B0C for 1 min. Cycles were repeated 40 times. Primers were =
as follows:=20
      ASPM, Quantitect Primers Assay (ref: QT00020209 Qiagen). The =
amount of=20
      cDNA was normalized to the amount of the <EM>ALAS =
</EM>housekeeping gene=20
      (Human Erythroid 5-Aminolevulinate Synthase) using primers F:=20
      5'-TGCAGTCCTCAGCGCAGTCT-3' and R: 5'-TGGCCCCAACTTCCATCAT-3'. The=20
      2<SUP>-=CE=94=CE=94CT </SUP>equation was applied to calculate the =
relative copy=20
      number of tumor samples <EM>versus </EM>non-neoplastic=20
tissues.</DIV></DIV>
      <DIV class=3Dsec id=3D__secid452848>
      <DIV class=3D"head2 head-separate">Cell Isolation and =
culture</DIV>
      <DIV class=3D"p p-first" id=3D__pid452852>Briefly, tumors were =
immediately=20
      collected after surgery in HBSS (14175, Invitrogen) and dissected =
after=20
      removal of gross necrosis, subjected to a trypsin digestion (5 =
mg/mL) for=20
      10 min at 37=C2=B0C, and then trituration and filtering. The cell =
suspension=20
      obtained was then plated at a density of 5.10<SUP>5 </SUP>cells/75 =

      cm<SUP>2 </SUP>flask (Corning, FRANCE) in 10 mL of DMEM/F12 medium =

      supplemented with Insulin (5 mg/mL), Penicillin - Streptomycin (50 =
mg/mL)=20
      and growth factor (bFGF; 20 ng/mL), epidermal growth factor (EGF; =
20=20
      ng/mL) and B27 (1/50) as a stem cell-permissive medium and =
maintained in a=20
      5.0% CO2/95% O2 atmosphere with 100% humidity. Cultures were fed =
every=20
      three days by changing half of the medium. Primary gliomaspheres =
were=20
      obtained after 5-10 days.</DIV>
      <DIV class=3D"p p-last" id=3D__pid452869>The U87 and U251 =
glioblastoma cell=20
      lines and the HEK293T kidney cell line were purchased from ATCC =
(American=20
      Type Culture Collection, USA), grown on DMEM 10% FCS- 1%=20
      Penicillin-Streptomycin, and used as controls for <EM>in vitro=20
      </EM>experiments.</DIV></DIV>
      <DIV class=3Dsec id=3D__secid452877>
      <DIV class=3D"head2 head-separate">LV-shRNA transduction and =
viability=20
      Fluorescence-activated cell sorter analysis of Gliomaspheres</DIV>
      <DIV class=3D"p p-first" id=3D__pid452882>We used pGIPZ (RHS4287, =
Biovalley=20
      FRANCE) lentiviral vectors expressing a short hairpin RNA (LV- =
shRNAmir,=20
      Biovalley, FRANCE) for ASPM silencing (sequence:=20
      =
TGCTGTTGACAGTGAGCGCGGCATTGGCGTGCTTATTTAATAGTGAAGCCACAGATGTATTAAATAAGCACGC=
CAATGCCTTGCCTACTGCCTCGGA=20
      or a scrambled non-targeted shRNA (RHS 4346; Biovalley FRANCE.) as =
a=20
      control.</DIV>
      <DIV class=3Dp id=3D__pid452887>Lentiviral vector stocks were =
produced by=20
      transient transfection of HEK293T cells with the p8.91 =
encapsidation=20
      plasmid [<A class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/9306402"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B9">9</A>], the VSV glycoprotein-G-encoding pHCMV-G plasmid =
[<A=20
      class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/7937806"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B10">10</A>], and the lentiviral recombinant vector as =
previously=20
      described [<A class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/10786833"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B11">11</A>]. Supernatants were treated with DNAse I (Roche =

      Diagnostic) prior to ultracentrifugation, and the resulting pellet =
was=20
      resuspended in Phosphate Buffered Saline, separated into aliquots =
and=20
      frozen at -80=C2=B0C until use. The transduction efficiency of =
each vector=20
      stock was determined by FACS analysis as previously described [<A=20
      class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/15759109"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B12">12</A>].</DIV>
      <DIV class=3D"p p-last" id=3D__pid828813>Gliomasphere cultures =
were=20
      mechanically disaggregated in order to increase the efficiency and =

      uniformity of infection and then transduced at 3-5.10<SUP>6 =
</SUP>cell/mL=20
      with LV-sh-miR of ASPM and non-targeted shRNA at a multiplicity of =

      infection of 2.5 (MOI: number of transducing units per cell) =
supplemented=20
      with DEAE dextran (20 =CE=BCg/mL). After expansion of the culture, =
GFP-positive=20
      viable cells were separated by cell sorting and flow cytometry =
FACS (FACS=20
      Aria=E2=84=A2 PQ1300010, BD Biosciences) to ensure knockdown =
efficiency. After 7=20
      days, ASPM RNA expression was measured in glioma stem cells =
infected with=20
      lentiviruses by real-time qPCR.</DIV></DIV>
      <DIV class=3Dsec id=3D__secid828825>
      <DIV class=3D"head2 head-separate">Proliferation and cell death=20
      quantification</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid828829>Proliferation was =
assessed by=20
      EDU incorporation (Click iT=E2=84=A2 EdU Flow Cytometry Assay =
Kits, A10202=20
      Invitrogen), and cell death by 7-AAD<SUP>+</SUP>cell staining. The =

      percentage of labeled cells was quantified by cell sorting on flow =

      cytometry FACS (FACS Aria=E2=84=A2 PQ1300010, BD =
Biosciences).</DIV></DIV>
      <DIV class=3Dsec id=3D__secid828838>
      <DIV class=3D"head2 head-separate">Subcutaneous Xenograft =
formation=20
      assay</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid828842>2 to 5 =C3=97 =
10<SUP>5 </SUP>cells=20
      suspended in 100 =CE=BCl of HBSS medium were injected =
subcutaneously into=20
      <EM>nude </EM>mice. Before xenograft establishment, tumors were =
passed=20
      four times and genomic stability was checked by CGH array=20
      analysis.</DIV></DIV>
      <DIV class=3Dsec id=3D__secid828854>
      <DIV class=3D"head2 head-separate">ASPM Immunofluorescence</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid828858>To detect the =
endogenous ASPM=20
      protein, cultured cells were fixed for 15 min in 4% =
paraformaldehyde,=20
      washed twice in PBS, treated for 30 min with 0.1 Triton X-100 -5% =
FCS, and=20
      incubated for 30 minutes at room temperature with Rabbit-anti-ASPM =
(IHC=20
      Antibody Affinity purified Cat IHC_00058 BETHYL, dilution 1:50) in =
0.1=20
      Triton X-100 -1% FCS. After two washes in 0.1 Triton X-100 -1% =
FCS,=20
      samples were incubated for 30 minutes at room temperature with =
goat=20
      anti-rabbit IgG-FITC (dilution 1:1000) in 0.1 Triton X-100 -1% =
FCS. After=20
      two washes, samples were stained with Hoechst solution and rinsed =
with=20
      PBS. Samples were mounted with Fluoromount<SUP>=C2=AE =
</SUP>(0100-01,=20
      SouthernBiotech, France). Images were captured with a Zeiss =
microscope=20
      (Axiophoto) equipped with a cooled CCD camera (Cool SNAP HQ, =
Photometrics,=20
      Tucson, AZ, USA and analyzed by IPLab software).</DIV></DIV>
      <DIV class=3Dsec id=3D__secid828872>
      <DIV class=3D"head2 head-separate">Protein extraction and ASPM =
Western=20
      blot</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid828876>Total protein, =
obtained by=20
      lysing tumor tissue or cells with RIPA 1=C3=97 buffer (SDS 20%, =
NP-40 20%,=20
      Deoxycholate acid 10%, Tris-HCl 2 M pH: 8, NaCl 5 M for 10=C3=97 =
buffer) was=20
      quantified using a Pierce BCA=E2=84=A2 Protein Assay Kit (ref =
23225 Pierce, USA),=20
      separated on Nu-Page 3-8% Tris-acetate polyacrylamide gels =
(Invitrogen),=20
      and electro-transferred to nitrocellulose membranes (Bio-Rad =
Laboratories,=20
      Inc.) in 25 mM Tris, 192 mM glycine, 20% ethanol, and 0.01% SDS =
for 2.5 -=20
      3 hours at 80-150 volts. After blocking in 5% milk solution, the =
membranes=20
      were probed with anti- ASPM (K-17 sc-48883) and anti- GAPDH (V-18=20
      sc-20357) both from Santa Cruz Biotechnology, Inc., U.S.A. =
Antibodies were=20
      diluted 1:500 for ASPM and 1:700 for GAPDH in TBST (50 mM Tris =
Base, 150=20
      mM NaCl, 0.1% (v/v) Tween-20) containing 5% milk powder. After =
washing in=20
      TBST, the membrane was incubated with appropriate horseradish =
peroxidase=20
      (HRP)-conjugated secondary antibodies diluted 1:5000 in TBST. The=20
      immunoreactive signals were detected using an ECL reagent Advance =
Western=20
      Blotting Detection Kit (GE Healthcare) and an Image Reader =
<EM>LAS-1000=20
      Pro ver. 2 </EM>(KODAK PHOTO FILM 4000 mm, FRANCE).</DIV></DIV>
      <DIV class=3D"sec sec-last" id=3D__secid828892>
      <DIV class=3D"head2 head-separate">Statistical analysis</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid828896>The =CF=87<SUP>2 =
</SUP>test was used=20
      to compare qualitative variable distribution, and the Mann-Whitney =
test=20
      was used for continuous variables. A two-sided p-value &lt; 0.05 =
was=20
      considered to be significant.</DIV></DIV></DIV></DIV>
      <DIV class=3Dsec id=3D__secid828904>
      <DIV class=3D"head1 section-title" id=3D__secid828904titletitle=20
      style=3D"TEXT-TRANSFORM: none">
      <DIV class=3Dother-sections>
      <UL class=3D"noext-menu noext-menu-hidden">
        <LI><A class=3Dfirst-link=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">&nbsp;Other=20
        Sections=E2=96=BC</A>
        <UL class=3D"submenu head1">
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__abstractid683998">Abstract</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid684043">Introduction</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid834380">Materials=20
          and methods</A>
          <LI class=3D"submenu-item current-item"><A class=3D""=20
          style=3D"TEXT-TRANSFORM: none" onclick=3Dreturn(false)=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">Results</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid704257">Discussion</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839339">Competing=20
          interests</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839346">Authors'=20
          contributions</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839355">Supplementary=20
          Material</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__ref-listid731707">References</A></LI></UL></LI></UL></DIV>
      <DIV>Results</DIV></DIV>
      <DIV class=3Dsection-content id=3D__secid828904content>
      <DIV class=3D"sec sec-first" id=3D__secid828908>
      <DIV class=3D"head2 head-separate">1. ASPM expression increases =
with glioma=20
      grade</DIV>
      <DIV class=3D"p p-first" id=3D__pid828912>We measured ASPM =
expression via=20
      qRT-PCR in 175 gliomas (8 Grade II, 75 Grade III and 92 Grade IV) =
and in=20
      three non-neoplastic brain tissues (NT). ASPM expression level =
increased=20
      between NT tissue and Grade II gliomas (p &lt; 0.05), between =
Grade II and=20
      Grade III (p &lt; 0.005), and between Grade III and Grade IV (p =
=3D 0.0001).=20
      ASPM expression did not differ between astrocytic (2 AII, 36 =
AIII),=20
      oligodendroglial (1 OII, 13 OIII) or mixed glioma samples (5 OAII, =
26 OA=20
      III) (Figure <A class=3D"fig-table-link fig figpopup"=20
      style=3D"TEXT-DECORATION: none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F1/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3.5em">(Figure1a</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">1a</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 1"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 1"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-1.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-1.jpg"></SPAN></SPAN=
></A>).</DIV>
      <DIV class=3Dcanvas-figure-ref-outer>
      <DIV class=3Dcanvas-figure-ref-inner><A id=3DF1 name=3DF1></A>
      <TABLE class=3Dthumb-caption style=3D"CLEAR: both; WIDTH: 100%" =
cellSpacing=3D0=20
      cellPadding=3D0 border=3D0>
        <TBODY>
        <TR vAlign=3Dtop align=3Dleft>
          <TD class=3Dthumb-cell><A class=3D"icon-reflink figpopup"=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F1/">
            <DIV class=3Dthumb-ph><SPAN class=3Dlarge-thumb-canvas><SPAN =

            class=3Dlarge-thumb-canvas-1><IMG class=3D"icon-reflink =
large-thumb"=20
            title=3D"Figure 1"=20
            style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
            alt=3D"Figure 1"=20
            =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-1.gif"=20
            =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-1.jpg"></SPAN></SPAN=
>
            <DIV class=3Dsmall-thumb-canvas>
            <DIV class=3Dsmall-thumb-canvas-1><IMG=20
            class=3D"icon-reflink small-thumb" title=3D"Figure 1" =
alt=3D"Figure 1"=20
            =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-1.gif"></DIV></DIV></DIV></A></TD>
          <TD class=3Dcaption-cell>
            <DIV class=3Dcaption-ph><A class=3Dside-caption=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F1/"><=
STRONG>Figure=20
            1</STRONG></A>
            <DIV =
class=3Dfigure-table-caption-in-article><SPAN><STRONG><EM>Aspm=20
            </EM>expression correlates with glioma grade and increases =
at=20
            recurrence</STRONG>. <STRONG>(a) </STRONG><EM>Aspm </EM>is=20
            up-regulated in gliomas compared to non-tumor brain and =
increases=20
            with the tumor grade;<STRONG>(b) ASPM </STRONG>expression =
increases=20
            between the initial tumor and recurrence at a =
higher</SPAN><A=20
            class=3Dside-caption style=3D"FONT-SIZE: 100%"=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F1/"> =

            (more ...)</A></DIV>
            <DIV=20
      =
class=3Dfigure-table-caption-in-article></DIV></DIV></TD></TR></TBODY></T=
ABLE>
      <DIV class=3Dfigure-table-caption-in-article></DIV></DIV></DIV>
      <DIV style=3D"CLEAR: both"></DIV>
      <DIV class=3D"p p-last" id=3D__pid740702>Preliminary results with =
western blot=20
      analysis showed that ASPM was also expressed at the protein level =
in=20
      gliomas and overexpressed in Grade IV gliomas as compared to =
Grades III=20
      and II (additional file <A class=3D"side-supplink =
supplementary-material"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#S1"=20
      rid=3D"S1">1</A>, Figure S1).</DIV></DIV>
      <DIV class=3Dsec id=3D__secid740716>
      <DIV class=3D"head2 head-separate">2. ASPM is upregulated in =
recurrent=20
      gliomas relative to the initial tumor</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid740721>ASPM mRNA =
expression was=20
      measured in 11 recurrent tumors and compared to the initial tumor =
(3 Grade=20
      II, 1 Grade III and 7 Grade IV). As shown in Figure <A=20
      class=3D"fig-table-link fig figpopup" style=3D"TEXT-DECORATION: =
none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F1/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3em">Figure1b,</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">1b</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 1"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 1"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-1.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-1.jpg"></SPAN></SPAN=
></A>,=20
      increased malignancy at recurrence was identified in all 4=20
      non-glioblastoma patients with concomitant increase of ASPM =
expression in=20
      all of them. In the 7 patients whose initial tumor was a GBM =
(Figure <A=20
      class=3D"fig-table-link fig figpopup" style=3D"TEXT-DECORATION: =
none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F1/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3.5em">(Figure1c),</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">1c</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 1"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 1"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-1.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-1.jpg"></SPAN></SPAN=
></A>),=20
      ASPM expression was 3.5-fold higher in the recurrent tumor =
although the=20
      grade did not change (p &lt; 0.005).</DIV></DIV>
      <DIV class=3Dsec id=3D__secid740747>
      <DIV class=3D"head2 head-separate">3. ASPM expression increases =
with the=20
      number of passages in gliomaspheres and in xenografted =
tumors</DIV>
      <DIV class=3D"p p-first" id=3D__pid740752>Because ASPM expression =
increases=20
      with the recurrence of glioma, and because ASPM is involved in =
tumor stem=20
      cells, we investigated ASPM expression over time <EM>in vitro =
</EM>in=20
      spheroid cultures and <EM>in vivo </EM>on xenografts</DIV>
      <DIV class=3Dp id=3D__pid740764>We generated three gliomaspheres =
cultures,=20
      GBM1, GBM2, GBM3, all characterized by EGFR amplification and 10q =
loss on=20
      CGH-array analysis; this genetic profile remained stable =
throughout the=20
      passages (as shown in additional file <A=20
      class=3D"side-supplink supplementary-material"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#S2"=20
      rid=3D"S2">2</A>, Figure S2 for GBM1). We also verified the =
capacity of=20
      gliomaspheres to differentiate in the presence of fetal bovine =
serum and=20
      to express multilineage markers (additional file 2, Figure S2). =
After=20
      dissociation of gliomaspheres, ASPM was detected during mitosis =
and=20
      located at the spindle poles of dividing cells (Figure <A=20
      class=3D"fig-table-link fig figpopup" style=3D"TEXT-DECORATION: =
none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F2/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3.5em">(Figure2a).</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">2a</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 2"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 2"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-2.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-2.jpg"></SPAN></SPAN=
></A>).=20
      ASPM transcript expression significantly increased throughout the =
passages=20
      for all gliomaspheres studied (Figure <A=20
      class=3D"fig-table-link fig figpopup" style=3D"TEXT-DECORATION: =
none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F2/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3.5em">(Figure2b).</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">2b</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 2"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 2"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-2.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-2.jpg"></SPAN></SPAN=
></A>).=20
      For example, in GBM1 gliomaspheres, we observed a 6.5-fold =
increase in=20
      ASPM mRNA level between passages 8 and 28.</DIV>
      <DIV class=3Dcanvas-figure-ref-outer>
      <DIV class=3Dcanvas-figure-ref-inner><A id=3DF2 name=3DF2></A>
      <TABLE class=3Dthumb-caption style=3D"CLEAR: both; WIDTH: 100%" =
cellSpacing=3D0=20
      cellPadding=3D0 border=3D0>
        <TBODY>
        <TR vAlign=3Dtop align=3Dleft>
          <TD class=3Dthumb-cell><A class=3D"icon-reflink figpopup"=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F2/">
            <DIV class=3Dthumb-ph><SPAN class=3Dlarge-thumb-canvas><SPAN =

            class=3Dlarge-thumb-canvas-1><IMG class=3D"icon-reflink =
large-thumb"=20
            title=3D"Figure 2"=20
            style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
            alt=3D"Figure 2"=20
            =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-2.gif"=20
            =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-2.jpg"></SPAN></SPAN=
>
            <DIV class=3Dsmall-thumb-canvas>
            <DIV class=3Dsmall-thumb-canvas-1><IMG=20
            class=3D"icon-reflink small-thumb" title=3D"Figure 2" =
alt=3D"Figure 2"=20
            =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-2.gif"></DIV></DIV></DIV></A></TD>
          <TD class=3Dcaption-cell>
            <DIV class=3Dcaption-ph><A class=3Dside-caption=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F2/"><=
STRONG>Figure=20
            2</STRONG></A>
            <DIV =
class=3Dfigure-table-caption-in-article><SPAN><STRONG>ASPM=20
            expression in gliomaspheres increases with successive =
passages=20
            <EM>in vitro </EM>and <EM>in vivo</EM></STRONG>. <STRONG>(a) =

            </STRONG>Metaphase staining of ASPM protein at both poles of =
the=20
            spindle. ASPM protein (green) is detected in gliomaspheres =
(arrow).=20
            Nuclei are stained with DAPI (blue);</SPAN><A =
class=3Dside-caption=20
            style=3D"FONT-SIZE: 100%"=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F2/"> =

            (more ...)</A></DIV>
            <DIV=20
      =
class=3Dfigure-table-caption-in-article></DIV></DIV></TD></TR></TBODY></T=
ABLE>
      <DIV class=3Dfigure-table-caption-in-article></DIV></DIV></DIV>
      <DIV style=3D"CLEAR: both"></DIV>
      <DIV class=3D"p p-last" id=3D__pid752526>After four passages and =
after=20
      checking genomic stability via CGH-a (not shown), 500,000 =
dissociated=20
      cells from GBM1 gliomaspheres were injected subcutaneously into=20
      immunodeficient nude mice. ASPM mRNA, measured by RT-PCR, =
increased=20
      progressively with successive tumor passages (Figure <A=20
      class=3D"fig-table-link fig figpopup" style=3D"TEXT-DECORATION: =
none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F2/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3.5em">(Figure2c</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">2c</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 2"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 2"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-2.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-2.jpg"></SPAN></SPAN=
></A>).</DIV></DIV>
      <DIV class=3Dsec id=3D__secid752540>
      <DIV class=3D"head2 head-separate">4. ASPM silencing inhibits cell =

      proliferation/gliomasphere formation and promotes cell death</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid733580>To determine the =
role of ASPM,=20
      we infected gliomaspheres with lentivirus expressing a sh-miR-RNA =
that=20
      specifically target <EM>Aspm</EM>. In GBM1, GBM2 and GBM3 =
gliomasphere=20
      models, ~80% of cells were successfully transduced with the =
sh-miR-RNA=20
      specific to <EM>Aspm</EM>. In transduced cells, ASPM expression =
measured=20
      by RT-PCR was dramatically reduced to less than 5% in GBM2 and =
less than=20
      0.02% in GBM1 and GBM3 (additional file <A=20
      class=3D"side-supplink supplementary-material"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#S3"=20
      rid=3D"S3">3</A>, Figure S3). We then analyzed cell kinetics in =
three=20
      different groups: non-transduced cells, cells transduced with the=20
      non-targeted sh-miR-RNA, and cells transduced with =
sh-miR-<EM>Aspm</EM>.=20
      After day 5, cells transduced with the non-silencing sh-miR-RNA=20
      proliferated again while the sh-miR-<EM>Aspm </EM>cells died =
(Figure <A=20
      class=3D"fig-table-link fig figpopup" style=3D"TEXT-DECORATION: =
none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F3/"><=
SPAN=20
      style=3D"TEXT-DECORATION: underline">3a-b</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 3"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 3"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-3.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-3.jpg"></SPAN></SPAN=
></A>).=20
      The morphology of the cells is shown at day 33 in Figure <A=20
      class=3D"fig-table-link fig figpopup" style=3D"TEXT-DECORATION: =
none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F3/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3em">Figure3c</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">3c</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 3"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 3"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-3.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-3.jpg"></SPAN></SPAN=
></A>=20
      and <A class=3D"fig-table-link fig figpopup" =
style=3D"TEXT-DECORATION: none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F3/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -1.5em">and3d.</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">3d</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 3"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 3"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-3.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-3.jpg"></SPAN></SPAN=
></A>.=20
      We then quantified the effects of <EM>Aspm </EM>silencing on =
proliferation=20
      (assessed by 5-Ethyl -2'-Deoxyuridine (EdU) incorporation) and =
cell death=20
      (assessed by 7-AAD<SUP>+ </SUP>labeling). We noted an accumulation =
of dead=20
      cells in sh-miR-<EM>Aspm </EM>populations (80% of =
7-AAD<SUP>+</SUP>GBM1,=20
      as shown in Figure <A class=3D"fig-table-link fig figpopup"=20
      style=3D"TEXT-DECORATION: none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F4/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3em">Figure4b,</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">4b</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 4"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 4"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-4.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-4.jpg"></SPAN></SPAN=
></A>,=20
      and 85% of 7-AAD<SUP>+ </SUP>GBM2) compared to control (Figure <A=20
      class=3D"fig-table-link fig figpopup" style=3D"TEXT-DECORATION: =
none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F4/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3.5em">(Figure4a).</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">4a</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 4"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 4"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-4.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-4.jpg"></SPAN></SPAN=
></A>).=20
      In addition to cell death, ASPM inhibition resulted in a decrease =
in=20
      proliferation: in sh-miR-<EM>Aspm </EM>GBM1 gliomaspheres, only =
25% of the=20
      viable cells (i.e., 5% of total cells) were EdU+ cells (Figure <A=20
      class=3D"fig-table-link fig figpopup" style=3D"TEXT-DECORATION: =
none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F4/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3.5em">(Figure4d),</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">4d</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 4"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 4"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-4.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-4.jpg"></SPAN></SPAN=
></A>),=20
      compared to 64% in the non-silenced cell population (Figure <A=20
      class=3D"fig-table-link fig figpopup" style=3D"TEXT-DECORATION: =
none"=20
      onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F4/"><=
SPAN=20
      style=3D"POSITION: relative; TEXT-DECORATION: none">=E2=80=8B<SPAN =

      class=3Dfigpopup-sensitive-area=20
      style=3D"LEFT: -3.5em">(Figure4c).</SPAN></SPAN><SPAN=20
      style=3D"TEXT-DECORATION: underline">4c</SPAN><SPAN=20
      class=3Dlarge-thumb-canvas><SPAN class=3Dlarge-thumb-canvas-1><IMG =

      class=3D"icon-reflink large-thumb" title=3D"Figure 4"=20
      style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
      alt=3D"Figure 4"=20
      =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-4.gif"=20
      =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-4.jpg"></SPAN></SPAN=
></A>).=20
      For GBM2, the results were similar: 4% of 15% of viable cells were =
EDU+ in=20
      sh-miR-<EM>Aspm </EM>GBM2 gliomaspheres <EM>vs</EM>. 81% in =
control GBM2=20
      (data not shown).</DIV>
      <DIV class=3Dcanvas-figure-ref-outer>
      <DIV class=3Dcanvas-figure-ref-inner><A id=3DF3 name=3DF3></A>
      <TABLE class=3Dthumb-caption style=3D"CLEAR: both; WIDTH: 100%" =
cellSpacing=3D0=20
      cellPadding=3D0 border=3D0>
        <TBODY>
        <TR vAlign=3Dtop align=3Dleft>
          <TD class=3Dthumb-cell><A class=3D"icon-reflink figpopup"=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F3/">
            <DIV class=3Dthumb-ph><SPAN class=3Dlarge-thumb-canvas><SPAN =

            class=3Dlarge-thumb-canvas-1><IMG class=3D"icon-reflink =
large-thumb"=20
            title=3D"Figure 3"=20
            style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
            alt=3D"Figure 3"=20
            =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-3.gif"=20
            =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-3.jpg"></SPAN></SPAN=
>
            <DIV class=3Dsmall-thumb-canvas>
            <DIV class=3Dsmall-thumb-canvas-1><IMG=20
            class=3D"icon-reflink small-thumb" title=3D"Figure 3" =
alt=3D"Figure 3"=20
            =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-3.gif"></DIV></DIV></DIV></A></TD>
          <TD class=3Dcaption-cell>
            <DIV class=3Dcaption-ph><A class=3Dside-caption=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F3/"><=
STRONG>Figure=20
            3</STRONG></A>
            <DIV =
class=3Dfigure-table-caption-in-article><SPAN><STRONG>ASPM=20
            knockdown in glioblastoma gliomaspheres</STRONG>. Cell =
kinetics of=20
            GBM1 <STRONG>(a) </STRONG>and GBM2 <STRONG>(b)=20
            </STRONG>gliomaspheres. The data were generated from three=20
            independent experiments. Gliomaspheres treated with =
sh-miR-<EM>Aspm=20
            </EM>do not grow as compared to non-transduced=20
            gliomaspheres</SPAN><A class=3Dside-caption =
style=3D"FONT-SIZE: 100%"=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F3/"> =

            (more ...)</A></DIV>
            <DIV=20
      =
class=3Dfigure-table-caption-in-article></DIV></DIV></TD></TR></TBODY></T=
ABLE>
      <DIV class=3Dfigure-table-caption-in-article></DIV></DIV></DIV>
      <DIV style=3D"CLEAR: both"></DIV>
      <DIV class=3Dcanvas-figure-ref-outer>
      <DIV class=3Dcanvas-figure-ref-inner><A id=3DF4 name=3DF4></A>
      <TABLE class=3Dthumb-caption style=3D"CLEAR: both; WIDTH: 100%" =
cellSpacing=3D0=20
      cellPadding=3D0 border=3D0>
        <TBODY>
        <TR vAlign=3Dtop align=3Dleft>
          <TD class=3Dthumb-cell><A class=3D"icon-reflink figpopup"=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F4/">
            <DIV class=3Dthumb-ph><SPAN class=3Dlarge-thumb-canvas><SPAN =

            class=3Dlarge-thumb-canvas-1><IMG class=3D"icon-reflink =
large-thumb"=20
            title=3D"Figure 4"=20
            style=3D"BORDER-RIGHT: 1px solid; BORDER-TOP: 1px solid; =
BORDER-LEFT: 1px solid; BORDER-BOTTOM: 1px solid"=20
            alt=3D"Figure 4"=20
            =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-4.gif"=20
            =
hires=3D"/pmc/articles/PMC2817685/bin/1475-2867-10-1-4.jpg"></SPAN></SPAN=
>
            <DIV class=3Dsmall-thumb-canvas>
            <DIV class=3Dsmall-thumb-canvas-1><IMG=20
            class=3D"icon-reflink small-thumb" title=3D"Figure 4" =
alt=3D"Figure 4"=20
            =
src=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867-=
10-1-4.gif"></DIV></DIV></DIV></A></TD>
          <TD class=3Dcaption-cell>
            <DIV class=3Dcaption-ph><A class=3Dside-caption=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F4/"><=
STRONG>Figure=20
            4</STRONG></A>
            <DIV =
class=3Dfigure-table-caption-in-article><SPAN><STRONG>Cell death=20
            and proliferation inhibition induced by ASPM silencing, =
evaluated by=20
            7-Actinomycin D (7-AAD) labeling and 5- Ethyl -2'- =
Deoxyuridine=20
            (EdU) incorporation</STRONG>. Compared to control =
<STRONG>(a),=20
            </STRONG>transduction of GBM1 with sh-miR-<EM>Aspm =
</EM>results in=20
            massive cell</SPAN><A class=3Dside-caption =
style=3D"FONT-SIZE: 100%"=20
            onclick=3D"startTarget(this, 'figure', 1024, 800)"=20
            =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/figure/F4/"> =

            (more ...)</A></DIV>
            <DIV=20
      =
class=3Dfigure-table-caption-in-article></DIV></DIV></TD></TR></TBODY></T=
ABLE>
      <DIV class=3Dfigure-table-caption-in-article></DIV></DIV></DIV>
      <DIV style=3D"CLEAR: both"></DIV></DIV>
      <DIV class=3D"sec sec-last" id=3D__secid704211>
      <DIV class=3D"head2 head-separate">5. ASPM silencing inhibited =
gliomasphere=20
      formation of GBM cancer stems cells</DIV>
      <DIV class=3D"p p-first-last" id=3D__pid704214>In order to =
evaluate more=20
      specifically the impact of <EM>Aspm </EM>silencing on GBM cancer =
stem=20
      cells, we conducted a secondary gliomasphere-forming assay in =
GBM1. Seven=20
      days after silencing, GFP<SUP>+ </SUP>cells were sorted and =
re-incubated=20
      at 1,000 cell/10 mL. We observed a dramatic decrease in cell =
number in=20
      GBM1 sh-miR-<EM>Aspm </EM>compared to the non-silenced control =
(1000 cells=20
      <EM>vs</EM>. 2.8.10<SUP>5 </SUP>cells at day 9 and 500 cells =
<EM>vs</EM>.=20
      3.7.10<SUP>6 </SUP>cells at day 15). At D15, no gliomaspheres were =

      observed (<EM>vs</EM>. 91 gliomaspheres in the control), =
indicating that=20
      the knockdown of <EM>Aspm </EM>completely abrogated the =
development of=20
      secondary gliomaspheres.</DIV></DIV></DIV></DIV>
      <DIV class=3Dsec id=3D__secid704257>
      <DIV class=3D"head1 section-title" id=3D__secid704257titletitle=20
      style=3D"TEXT-TRANSFORM: none">
      <DIV class=3Dother-sections>
      <UL class=3D"noext-menu noext-menu-hidden">
        <LI><A class=3Dfirst-link=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">&nbsp;Other=20
        Sections=E2=96=BC</A>
        <UL class=3D"submenu head1">
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__abstractid683998">Abstract</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid684043">Introduction</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid834380">Materials=20
          and methods</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid828904">Results</A>
          <LI class=3D"submenu-item current-item"><A class=3D""=20
          style=3D"TEXT-TRANSFORM: none" onclick=3Dreturn(false)=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">Discussion</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839339">Competing=20
          interests</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839346">Authors'=20
          contributions</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839355">Supplementary=20
          Material</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__ref-listid731707">References</A></LI></UL></LI></UL></DIV>
      <DIV>Discussion</DIV></DIV>
      <DIV class=3Dsection-content id=3D__secid704257content>
      <DIV class=3D"p p-first" id=3D__pid704261>This study indicates =
that ASPM is=20
      tightly associated with the malignant progression of gliomas and =
that its=20
      knockdown produces extensive death of glioma cells <EM>in=20
vitro</EM>.</DIV>
      <DIV class=3Dp id=3D__pid704269>As previously described [<A=20
      class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18636190"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B7">7</A>], we found that ASPM expression was strongly =
correlated=20
      with the grade of gliomas. In recurrent tumors, ASPM expression =
increased=20
      not only in patients who progressed to a higher grade, but also in =

      recurrent glioblastomas, suggesting that a progressive increase in =
ASPM=20
      mRNA expression is a marker for the inexorable malignant =
progression and=20
      aggressiveness of gliomas, continuing even when the tumors have =
already=20
      reached their most anaplastic histological stage (Grade IV). =
Experimental=20
      evidence supports this finding since we found a progressive =
increase in=20
      ASPM mRNA expression in serial passages of gliomaspheres <EM>in =
vitro=20
      </EM>and in mouse glioma xenografts <EM>in vivo</EM>. To explain =
such a=20
      tight correlation between ASPM expression and the malignant =
progression of=20
      tumors, an attractive hypothesis is that symmetric division of=20
      CSC-expressing ASPM leads to progressive proliferative expansion =
of this=20
      undifferentiated compartment. Although we found that ASPM was also =

      expressed at the protein level in all glioma grades and =
overexpressed in=20
      Grade IV gliomas, it is worth noting that the results were less =
striking.=20
      This finding has been previously noted and led to the suggestion =
that ASPM=20
      could be in part regulated at the translational level [<A=20
      class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18636190"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B7">7</A>]. However, the dramatic inhibition of =
gliomasphere=20
      proliferation obtained with shRNA-mediated ASPM knockdown =
highlights the=20
      potential importance of this gene in malignant gliomas. Sphere =
formation=20
      is a key behavior of neural and brain tumor stem cells and is used =
to test=20
      stem cell capacity for self-renewal. A previous study also showed =
that=20
      siRNA-mediated inhibition of ASPM resulted in cell proliferation=20
      inhibition, thought to be mediated by a G1-phase cell cycle arrest =
[<A=20
      class=3D"cite-reflink bibr popnode"=20
      href=3D"http://www.ncbi.nlm.nih.gov/pubmed/17090670"=20
      =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle%7CBody&amp;TO=3DEntrez%7CPubMed%7CRe=
cord&amp;rendering-type=3Dnormal"=20
      rid=3D"B6">6</A>]. The effect that we observed here by stable =
transduction=20
      of shRNA expression in CSC-enriched cells appears even more =
drastic,=20
      possibly because of a more efficient and longer lasting inhibition =
of=20
      <EM>ASPM</EM>. Whereas siRNA transfection results in a transient =
effect,=20
      lentiviral-mediated expression of shRNA allows the transgene to =
integrate=20
      into the genome of the host cell, resulting in long-term =
expression.=20
      Production of secondary spheres was also inhibited after ASPM =
knockdown, a=20
      finding compatible with a sustained inhibition of CSC =
proliferation.=20
      Interestingly, stable inhibition of ASPM not only prevented =
further growth=20
      but also produced rapid and extensive cellular apoptotic death in =
our=20
      model. This finding may indicate that, following ASPM inhibition, =
CSC=20
      underwent active maturation and differentiation but could not =
survive in=20
      our experimental setting (serum-free medium without appropriate =
growth=20
      factors). However, experiments by Horvath et <EM>al</EM>. =
conducted in a=20
      similar serum-free medium resulted in growth inhibition and not =
massive=20
      cell death. On the other hand, the function of <EM>ASPM </EM>in =
neoplastic=20
      cells, particularly gliomas, has not been completely elucidated. =
In=20
      particular, the links between ASPM and key molecular alterations =
of=20
      malignant gliomas, such as EGFR amplification or mutation =
(EGFRvIII) (an=20
      almost constant feature in gliomaspheres - 80% in our experience), =
remain=20
      to be investigated. Nevertheless, some data suggest that these =
alterations=20
      are closely related. ASPM mRNA expression was increased in an=20
      EGFRvIII-expressing U87 glioblatoma cell line as compared to an =
isogenic=20
      U87 line, while ASPM expression was inhibited following the =
addition of=20
      erlotinib (an EGFR tyrosine kinase inhibitor) to the =
EGFRvIII-expressing=20
      U87 line. This observation led Horvath et <EM>al </EM>to suggest =
that ASPM=20
      (and genes from the same module) is involved downstream of the =
mutant=20
      EGFR. Thus, inhibition of ASPM could prove lethal by severely =
hampering=20
      one of the key signaling pathways of glioblastomas.</DIV>
      <DIV class=3D"p p-last" id=3D__pid839326>Because ASPM is absent or =
expressed=20
      at very low levels in normal brain, the considerable differential=20
      expression levels between GBM and normal parenchyma and the =
extreme <EM>in=20
      vitro </EM>sensitivity of tumor cells to ASPM knockdown support =
continued=20
      research to specifically target ASPM from a therapeutic =
perspective,=20
      focusing on <EM>in vivo </EM>studies.</DIV></DIV></DIV>
      <DIV class=3Dsec id=3D__secid839339>
      <DIV class=3D"head1 section-title" id=3D__secid839339titletitle=20
      style=3D"TEXT-TRANSFORM: none">
      <DIV>Competing interests</DIV></DIV>
      <DIV class=3Dsection-content id=3D__secid839339content>
      <DIV class=3D"p p-first-last" id=3D__pid839343>The authors declare =
that they=20
      have no competing interests.</DIV></DIV></DIV>
      <DIV class=3Dsec id=3D__secid839346>
      <DIV class=3D"head1 section-title" id=3D__secid839346titletitle=20
      style=3D"TEXT-TRANSFORM: none">
      <DIV>Authors' contributions</DIV></DIV>
      <DIV class=3Dsection-content id=3D__secid839346content>
      <DIV class=3D"p p-first-last" id=3D__pid839350>SNNB, CC and YM =
carried out=20
      most of the in vitro work. RV and PR produced the lentivirus. =
SNNB. AR and=20
      BB performed the immunohistochemistry experiments. AI performed =
the=20
      CGH-array experiments. PL and ML did the in vivo experiments. SEH=20
      contributed to the in vitro experiments. MS conceived the study. =
JYD and=20
      MS wrote the manuscript. All authors read and approved the final=20
      manuscript.</DIV></DIV></DIV>
      <DIV class=3Dsec id=3D__secid839355>
      <DIV class=3D"head1 section-title" id=3D__secid839355titletitle=20
      style=3D"TEXT-TRANSFORM: none">
      <DIV class=3Dother-sections>
      <UL class=3D"noext-menu noext-menu-hidden">
        <LI><A class=3Dfirst-link=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">&nbsp;Other=20
        Sections=E2=96=BC</A>
        <UL class=3D"submenu head1">
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__abstractid683998">Abstract</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid684043">Introduction</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid834380">Materials=20
          and methods</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid828904">Results</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid704257">Discussion</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839339">Competing=20
          interests</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid839346">Authors'=20
          contributions</A>
          <LI class=3D"submenu-item current-item"><A class=3D""=20
          style=3D"TEXT-TRANSFORM: none" onclick=3Dreturn(false)=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">Supplementary=20
          Material</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__ref-listid731707">References</A></LI></UL></LI></UL></DIV>
      <DIV><A id=3Dsupplementary-material-sec></A>Supplementary=20
      Material</DIV></DIV>
      <DIV class=3Dsection-content id=3D__secid839355content><!--body-->
      <DIV class=3Dp id=3DS1>
      <DIV class=3Dtitle><STRONG>Additional file 1</STRONG></DIV>
      <DIV class=3Dp id=3D__pid839375><STRONG>Figure S1 - ASPM protein=20
      expression</STRONG>. ASPM western blot analysis of grade II, III =
and IV=20
      gliomas (n =3D 4 in each group). Relative protein expression is =
presented as=20
      the ratio of density value for ASPM over GAPDH signal (mean +/-=20
SEM).</DIV>
      <DIV class=3Dsupplementary-material-media-label-caption><A =
class=3Dint-reflink=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867=
-10-1-S1.PPT">Click=20
      here for file</A><SUP>(185K, PPT)</SUP></DIV></DIV>
      <DIV class=3Dp id=3DS2>
      <DIV class=3Dtitle><STRONG>Additional file 2</STRONG></DIV>
      <DIV class=3Dp id=3D__pid731638><STRONG>Figure S2 - Tumor spheroid =

      characterization</STRONG>. Genomic stability was examined with =
CGHa=20
      analysis (left =3D DNA profile from initial tumor; right =3D DNA =
profile from=20
      gliomasphere at passage p28). The chromosomes are indicated on the =
x axis=20
      and copy number is on y axis. Yellow indicates the normal genomic =
copy=20
      number, while green indicates a loss and red indicates a gain in =
copy=20
      number. As shown here, GBM1 carries EGFR amplification and loss of =

      chromosome 10q. After CSC isolation in stem cell-permissive =
medium,=20
      gliomaspheres were differentiated in presence of fetal bovine =
serum and=20
      stained by Sox2 and O4 for stemness markers and by GFAP and Tuji1 =
to=20
      characterize the potential of multi lineage differentiation of =
GBM1.</DIV>
      <DIV class=3Dsupplementary-material-media-label-caption><A =
class=3Dint-reflink=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867=
-10-1-S2.PPT">Click=20
      here for file</A><SUP>(889K, PPT)</SUP></DIV></DIV>
      <DIV class=3Dp id=3DS3>
      <DIV class=3Dtitle><STRONG>Additional file 3</STRONG></DIV>
      <DIV class=3Dp id=3D__pid731676><STRONG>Figure S3 - Knock-down of =
Aspm=20
      gene</STRONG>. Tumor spheroids (GBM1, GBM2, GBM3) were transfected =
either=20
      with non silencing (scrambled ShRNA) or with ASPM ShRNA expressing =

      lentivirus, resulting in a dramatic drop of ASPM RNA in GBM1, GBM3 =
and to=20
      a lesser extent GBM2.</DIV>
      <DIV class=3Dsupplementary-material-media-label-caption><A =
class=3Dint-reflink=20
      =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/bin/1475-2867=
-10-1-S3.PPT">Click=20
      here for file</A><SUP>(109K, PPT)</SUP></DIV></DIV></DIV></DIV>
      <DIV class=3Dsec id=3D__secid731698>
      <DIV class=3D"head1 section-title" id=3D__secid731698titletitle=20
      style=3D"TEXT-TRANSFORM: none">
      <DIV>Acknowledgements</DIV></DIV>
      <DIV class=3Dsection-content id=3D__secid731698content><!--back-->
      <DIV class=3D"p p-first-last" id=3D__pid731702>This work was =
supported by the=20
      Institut National du Cancer (PL 046) and the Ligue Nationale =
contre le=20
      Cancer. SNNB is supported by a grant from the ARTC (Association =
pour la=20
      Recherche des Tumeurs C=C3=A9r=C3=A9brales).</DIV></DIV></DIV>
      <DIV class=3Dsec id=3D__ref-listid731707>
      <DIV class=3D"head1 section-title" =
id=3D__ref-listid731707titletitle=20
      style=3D"TEXT-TRANSFORM: none">
      <DIV class=3Dother-sections>
      <UL class=3D"noext-menu noext-menu-hidden">
        <LI><A class=3Dfirst-link=20
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href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#">&nbsp;Other=20
        Sections=E2=96=BC</A>
        <UL class=3D"submenu head1">
          <LI class=3Dsubmenu-item><A class=3D"" =
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          =
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d#__abstractid683998">Abstract</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid684043">Introduction</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
style=3D"TEXT-TRANSFORM: none"=20
          =
href=3D"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817685/?tool=3Dpubme=
d#__secid834380">Materials=20
          and methods</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
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          =
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          <LI class=3Dsubmenu-item><A class=3D"" =
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          contributions</A>
          <LI class=3Dsubmenu-item><A class=3D"" =
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d#">References</A></LI></UL></LI></UL></DIV>
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rch|Author%20Search&amp;rendering-type=3Dnormal">Marie,=20
        Y.</A> </LI>
        <LI class=3Ditem><A=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=3Dsearch&amp;db=3DPu=
bMed&amp;term=3D%20Sanson%2BM[auth]"=20
        =
ref=3D"reftype=3Dauthsrch&amp;article-id=3D2817685&amp;issue-id=3D184705&=
amp;journal-id=3D96&amp;FROM=3DArticle|ByTheseAuthors&amp;TO=3DEntrez|Sea=
rch|Author%20Search&amp;rendering-type=3Dnormal">Sanson,=20
        M.</A> </LI></UL></DIV></DIV></DIV>
      <DIV class=3D"port norender brieflink" id=3DNCBIPageSection=20
      config=3D"collapsed: false, closeable: false">
      <H2>PubMed related articles</H2>
      <DIV class=3DPPMCPubmedRA>
      <UL class=3Dpop1>
        <LI class=3D"item popnode note"><A=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18636190?ordinalpos=3D1&amp;it=
ool=3DPPMCLayout.PPMCAppController.PPMCArticlePage.PPMCPubmedRA&amp;linkp=
os=3D1"=20
        =
ref=3D"reftype=3Drelart&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|RelatedArticles&amp;TO=3DEntrez|Pubm=
ed|Record&amp;rendering-type=3Dnormal">Expression=20
        analysis of the autosomal recessive primary microcephaly genes =
MCPH1=20
        (microcephalin) and MCPH5 (ASPM, abnormal spindle-like, =
microcephaly=20
        associated) in human malignant gliomas.</A>=20
        <P class=3Ddesc><EM class=3Dcit>Oncol Rep. 2008 Aug; =
20(2):301-8. </EM></P>
        <P class=3Dnote>[Oncol Rep. 2008]</P></LI>
        <LI class=3D"item popnode note"><A=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18676753?ordinalpos=3D1&amp;it=
ool=3DPPMCLayout.PPMCAppController.PPMCArticlePage.PPMCPubmedRA&amp;linkp=
os=3D2"=20
        =
ref=3D"reftype=3Drelart&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|RelatedArticles&amp;TO=3DEntrez|Pubm=
ed|Record&amp;rendering-type=3Dnormal">ASPM=20
        is a novel marker for vascular invasion, early recurrence, and =
poor=20
        prognosis of hepatocellular carcinoma.</A>=20
        <P class=3Ddesc><EM class=3Dcit>Clin Cancer Res. 2008 Aug 1; =
14(15):4814-20.=20
        </EM></P>
        <P class=3Dnote>[Clin Cancer Res. 2008]</P></LI>
        <LI class=3D"item popnode note"><A=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18577219?ordinalpos=3D1&amp;it=
ool=3DPPMCLayout.PPMCAppController.PPMCArticlePage.PPMCPubmedRA&amp;linkp=
os=3D3"=20
        =
ref=3D"reftype=3Drelart&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|RelatedArticles&amp;TO=3DEntrez|Pubm=
ed|Record&amp;rendering-type=3Dnormal">miR-124=20
        and miR-137 inhibit proliferation of glioblastoma multiforme =
cells and=20
        induce differentiation of brain tumor stem cells.</A>=20
        <P class=3Ddesc><EM class=3Dcit>BMC Med. 2008 Jun 24; 6:14. Epub =
2008 Jun=20
        24.</EM> </P>
        <P class=3Dnote>[BMC Med. 2008]</P></LI>
        <LI class=3D"item popnode note"><A=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pubmed/17005465?ordinalpos=3D1&amp;it=
ool=3DPPMCLayout.PPMCAppController.PPMCArticlePage.PPMCPubmedRA&amp;linkp=
os=3D4"=20
        =
ref=3D"reftype=3Drelart&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|RelatedArticles&amp;TO=3DEntrez|Pubm=
ed|Record&amp;rendering-type=3Dnormal"><SPAN=20
        class=3Dreviewflag>Review</SPAN>Molecular biology of malignant=20
        gliomas.</A>=20
        <P class=3Ddesc><EM class=3Dcit>Clin Transl Oncol. 2006 Sep; =
8(9):635-41.=20
        </EM></P>
        <P class=3Dnote>[Clin Transl Oncol. 2006]</P></LI>
        <LI class=3D"item popnode note"><A=20
        =
href=3D"http://www.ncbi.nlm.nih.gov/pubmed/19368079?ordinalpos=3D1&amp;it=
ool=3DPPMCLayout.PPMCAppController.PPMCArticlePage.PPMCPubmedRA&amp;linkp=
os=3D5"=20
        =
ref=3D"reftype=3Drelart&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|RelatedArticles&amp;TO=3DEntrez|Pubm=
ed|Record&amp;rendering-type=3Dnormal"><SPAN=20
        class=3Dreviewflag>Review</SPAN>Present and potential future =
adjuvant=20
        issues in high-grade astrocytic glioma treatment.</A>=20
        <P class=3Ddesc><EM class=3Dcit>Adv Tech Stand Neurosurg. 2009; =
34:3-35.=20
        </EM></P>
        <P class=3Dnote>[Adv Tech Stand Neurosurg. 2009]</P>
        <LI class=3Dmore>=C2=BB <A=20
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        Tree</A> </LI></UL></DIV></DIV></DIV></TD></TR></TBODY></TABLE>
<DIV class=3DArticleRefDocsums id=3DArticleRefDocsums>
<DIV class=3Dcited-ref id=3Dcite-__pid684047 rid=3D"__pid684047"=20
pmids=3D"15466194 15549107">
<UL>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/15466194"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">Isolation=20
  and characterization of tumorigenic, stem-like neural precursors from =
human=20
  glioblastoma.</A>
  <P class=3Ddesc><EM class=3Dcit>Cancer Res. 2004 Oct 1;=20
  64(19):7011-21.</EM></P><SPAN class=3Dpub>[Cancer Res. =
2004]</SPAN></LI>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/15549107"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">Identification=20
  of human brain tumour initiating cells.</A>
  <P class=3Ddesc><EM class=3Dcit>Nature. 2004 Nov 18;=20
  432(7015):396-401.</EM></P><SPAN class=3Dpub>[Nature. =
2004]</SPAN></LI></UL><A=20
class=3Dall-articles=20
href=3D"http://www.ncbi.nlm.nih.gov/pubmed/15466194,15549107/?report=3Dsu=
mmary"=20
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">See=20
more articles cited in this paragraph</A></DIV>
<DIV class=3Dcited-ref id=3Dcite-__pid684075 rid=3D"__pid684075"=20
pmids=3D"16798874 12355089 15972725 17090670 18636190">
<UL>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/16798874"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">Aspm=20
  specifically maintains symmetric proliferative divisions of =
neuroepithelial=20
  cells.</A>
  <P class=3Ddesc><EM class=3Dcit>Proc Natl Acad Sci U S A. 2006 Jul 5;=20
  103(27):10438-43.</EM></P><SPAN class=3Dpub>[Proc Natl Acad Sci U S A. =

  2006]</SPAN></LI>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/12355089"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">ASPM=20
  is a major determinant of cerebral cortical size.</A>
  <P class=3Ddesc><EM class=3Dcit>Nat Genet. 2002 Oct; =
32(2):316-20.</EM></P><SPAN=20
  class=3Dpub>[Nat Genet. 2002]</SPAN></LI>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/15972725"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">The=20
  microcephaly ASPM gene is expressed in proliferating tissues and =
encodes for a=20
  mitotic spindle protein.</A>
  <P class=3Ddesc><EM class=3Dcit>Hum Mol Genet. 2005 Aug 1;=20
  14(15):2155-65.</EM></P><SPAN class=3Dpub>[Hum Mol Genet. =
2005]</SPAN></LI>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/17090670"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">Analysis=20
  of oncogenic signaling networks in glioblastoma identifies ASPM as a =
molecular=20
  target.</A>
  <P class=3Ddesc><EM class=3Dcit>Proc Natl Acad Sci U S A. 2006 Nov 14; =

  103(46):17402-7.</EM></P><SPAN class=3Dpub>[Proc Natl Acad Sci U S A.=20
  2006]</SPAN></LI>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18636190"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">Expression=20
  analysis of the autosomal recessive primary microcephaly genes MCPH1=20
  (microcephalin) and MCPH5 (ASPM, abnormal spindle-like, microcephaly=20
  associated) in human malignant gliomas.</A>
  <P class=3Ddesc><EM class=3Dcit>Oncol Rep. 2008 Aug; =
20(2):301-8.</EM></P><SPAN=20
  class=3Dpub>[Oncol Rep. 2008]</SPAN></LI></UL><A class=3Dall-articles=20
href=3D"http://www.ncbi.nlm.nih.gov/pubmed/16798874,12355089,15972725,170=
90670,18636190/?report=3Dsummary"=20
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">See=20
more articles cited in this paragraph</A></DIV>
<DIV class=3Dcited-ref id=3Dcite-__pid834390 rid=3D"__pid834390" =
pmids=3D"18076069">
<UL>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18076069"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">BAC=20
  array CGH distinguishes mutually exclusive alterations that define=20
  clinicogenetic subtypes of gliomas.</A>
  <P class=3Ddesc><EM class=3Dcit>Int J Cancer. 2008 Apr 15;=20
  122(8):1778-86.</EM></P><SPAN class=3Dpub>[Int J Cancer. =
2008]</SPAN></LI></UL><A=20
class=3Dall-articles=20
href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18076069/?report=3Dsummary"=20
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">See=20
more articles cited in this paragraph</A></DIV>
<DIV class=3Dcited-ref id=3Dcite-__pid452887 rid=3D"__pid452887"=20
pmids=3D"9306402 7937806 10786833 15759109">
<UL>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/9306402"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">Multiply=20
  attenuated lentiviral vector achieves efficient gene delivery in =
vivo.</A>
  <P class=3Ddesc><EM class=3Dcit>Nat Biotechnol. 1997 Sep;=20
  15(9):871-5.</EM></P><SPAN class=3Dpub>[Nat Biotechnol. =
1997]</SPAN></LI>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/7937806"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">A=20
  general method for the generation of high-titer, pantropic retroviral =
vectors:=20
  highly efficient infection of primary hepatocytes.</A>
  <P class=3Ddesc><EM class=3Dcit>Proc Natl Acad Sci U S A. 1994 Sep 27; =

  91(20):9564-8.</EM></P><SPAN class=3Dpub>[Proc Natl Acad Sci U S A.=20
  1994]</SPAN></LI>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/10786833"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">HIV-1=20
  genome nuclear import is mediated by a central DNA flap.</A>
  <P class=3Ddesc><EM class=3Dcit>Cell. 2000 Apr 14; =
101(2):173-85.</EM></P><SPAN=20
  class=3Dpub>[Cell. 2000]</SPAN></LI>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/15759109"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">Efficient=20
  restricted gene expression in beta cells by lentivirus-mediated gene =
transfer=20
  into pancreatic stem/progenitor cells.</A>
  <P class=3Ddesc><EM class=3Dcit>Diabetologia. 2005 Apr;=20
  48(4):709-19.</EM></P><SPAN class=3Dpub>[Diabetologia. =
2005]</SPAN></LI></UL><A=20
class=3Dall-articles=20
href=3D"http://www.ncbi.nlm.nih.gov/pubmed/9306402,7937806,10786833,15759=
109/?report=3Dsummary"=20
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">See=20
more articles cited in this paragraph</A></DIV>
<DIV class=3Dcited-ref id=3Dcite-__pid704269 rid=3D"__pid704269"=20
pmids=3D"18636190 17090670">
<UL>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18636190"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">Expression=20
  analysis of the autosomal recessive primary microcephaly genes MCPH1=20
  (microcephalin) and MCPH5 (ASPM, abnormal spindle-like, microcephaly=20
  associated) in human malignant gliomas.</A>
  <P class=3Ddesc><EM class=3Dcit>Oncol Rep. 2008 Aug; =
20(2):301-8.</EM></P><SPAN=20
  class=3Dpub>[Oncol Rep. 2008]</SPAN></LI>
  <LI class=3Dovfl><A class=3D"pl popnode"=20
  href=3D"http://www.ncbi.nlm.nih.gov/pubmed/17090670"=20
  =
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
|Record&amp;rendering-type=3Dnormal">Analysis=20
  of oncogenic signaling networks in glioblastoma identifies ASPM as a =
molecular=20
  target.</A>
  <P class=3Ddesc><EM class=3Dcit>Proc Natl Acad Sci U S A. 2006 Nov 14; =

  103(46):17402-7.</EM></P><SPAN class=3Dpub>[Proc Natl Acad Sci U S A.=20
  2006]</SPAN></LI></UL><A class=3Dall-articles=20
href=3D"http://www.ncbi.nlm.nih.gov/pubmed/18636190,17090670/?report=3Dsu=
mmary"=20
ref=3D"reftype=3Dpubmed&amp;article-id=3D2817685&amp;issue-id=3D184705&am=
p;journal-id=3D96&amp;FROM=3DArticle|CitedRefBlock&amp;TO=3DEntrez|Pubmed=
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0px; LINE-HEIGHT: 1; PADDING-TOP: 0px; LIST-STYLE-TYPE: none; =
TEXT-ALIGN: left
}
.noext-menu A {
	DISPLAY: block; CURSOR: pointer; TEXT-DECORATION: underline; _display: =
inline; outline: none
}
.noext-menu LI {
=09
}
.noext-menu A.first-link {
	FONT-SIZE: 0.65em; TEXT-TRANSFORM: none; WHITE-SPACE: nowrap; =
TEXT-DECORATION: none
}
.noext-menu A.first-link:hover {
	TEXT-DECORATION: none! important
}
UL.noext-menu LI SPAN.delta {
=09
}
.noext-menu LI UL.submenu {
	DISPLAY: none; FONT-SIZE: 0.75em; POSITION: absolute
}
.noext-menu LI UL UL {
	MARGIN: -1em 0px 0px 10em
}
UL.noext-menu LI.submenu-item UL {
	DISPLAY: none; POSITION: absolute
}
UL.noext-menu LI.active-item UL {
	LEFT: 3px
}
UL.noext-menu LI UL.opened {
	BORDER-RIGHT: transparent 1px solid; PADDING-RIGHT: 0.25em; BORDER-TOP: =
transparent 1px solid; MARGIN-TOP: 0.25em; MIN-WIDTH: 200px; DISPLAY: =
block; PADDING-LEFT: 0.25em; Z-INDEX: 5; RIGHT: -0.1em; MAX-WIDTH: =
300px; PADDING-BOTTOM: 0.25em; BORDER-LEFT: transparent 1px solid; ; =
WIDTH: expression(this.clientWidth > 300 ? "300px" : "200px"); =
PADDING-TOP: 0.25em; BORDER-BOTTOM: transparent 1px solid; TOP: 1.4em; =
_top: 1.6em
}
UL.noext-menu LI UL.opened LI {
	MARGIN-BOTTOM: 3px
}
UL.noext-menu LI UL.opened LI.active-item {
	PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; =
PADDING-TOP: 0px; BACKGROUND-COLOR: #aaaaaa
}
UL.opened LI.active-item A {
	COLOR: #ffffff
}
UL.noext-menu LI UL A {
	PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 2px; MARGIN: 0px =
3px; COLOR: #222222; PADDING-TOP: 2px; TEXT-DECORATION: none
}
.noext-menu-hidden {
	COLOR: transparent! important; BACKGROUND-COLOR: transparent! important
}
.noext-menu-hidden A.first-link {
	COLOR: transparent! important; BACKGROUND-COLOR: transparent! important
}

------=_NextPart_000_0000_01CAD35F.003A3910
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.ncbi.nlm.nih.gov/core/extjs/ext-2.2/resources/css/panel.css

.x-panel {
	BORDER-RIGHT: #99bbe8 0px solid; BORDER-TOP: #99bbe8 0px solid; =
BORDER-LEFT: #99bbe8 0px solid; BORDER-BOTTOM: #99bbe8 0px solid
}
.x-panel-header {
	BORDER-RIGHT: #99bbe8 1px solid; PADDING-RIGHT: 3px; BORDER-TOP: =
#99bbe8 1px solid; PADDING-LEFT: 5px; BACKGROUND: =
url(../images/default/panel/white-top-bottom.gif) repeat-x 0px -1px; =
PADDING-BOTTOM: 4px; FONT: bold 11px/15px =
tahoma,arial,verdana,sans-serif; OVERFLOW: hidden; BORDER-LEFT: #99bbe8 =
1px solid; COLOR: #15428b; PADDING-TOP: 5px; BORDER-BOTTOM: #99bbe8 1px =
solid; ZOOM: 1
}
.x-panel-body {
	BORDER-RIGHT: #99bbe8 1px solid; BORDER-TOP: #99bbe8 0px; BACKGROUND: =
white; OVERFLOW: hidden; BORDER-LEFT: #99bbe8 1px solid; BORDER-BOTTOM: =
#99bbe8 1px solid; POSITION: relative
}
.x-panel-bbar .x-toolbar {
	BORDER-RIGHT: #99bbe8 1px solid; PADDING-RIGHT: 2px; BORDER-TOP: =
#99bbe8 0px; PADDING-LEFT: 2px; PADDING-BOTTOM: 2px; OVERFLOW: hidden; =
BORDER-LEFT: #99bbe8 1px solid; PADDING-TOP: 2px; BORDER-BOTTOM: #99bbe8 =
1px solid
}
.x-panel-tbar .x-toolbar {
	BORDER-RIGHT: #99bbe8 1px solid; PADDING-RIGHT: 2px; BORDER-TOP: =
#99bbe8 0px; PADDING-LEFT: 2px; PADDING-BOTTOM: 2px; OVERFLOW: hidden; =
BORDER-LEFT: #99bbe8 1px solid; PADDING-TOP: 2px; BORDER-BOTTOM: #99bbe8 =
1px solid
}
.x-panel-tbar-noheader .x-toolbar {
	BORDER-TOP: #99bbe8 1px solid; BORDER-BOTTOM: 0px
}
.x-panel-mc .x-panel-tbar .x-toolbar {
	BORDER-TOP: #99bbe8 1px solid; BORDER-BOTTOM: 0px
}
.x-panel-body-noheader {
	BORDER-TOP: #99bbe8 1px solid
}
.x-panel-mc .x-panel-body {
	BORDER-TOP: #99bbe8 1px solid
}
.x-panel-header {
	OVERFLOW: hidden; ZOOM: 1
}
.x-panel-tl .x-panel-header {
	BORDER-RIGHT: 0px; PADDING-RIGHT: 0px; BORDER-TOP: 0px; PADDING-LEFT: =
0px; BACKGROUND: none transparent scroll repeat 0% 0%; PADDING-BOTTOM: =
4px; FONT: bold 11px tahoma,arial,verdana,sans-serif; BORDER-LEFT: 0px; =
COLOR: #15428b; PADDING-TOP: 5px; BORDER-BOTTOM: 0px
}
.x-panel-tl .x-panel-icon {
	BACKGROUND-POSITION: 0px 4px; PADDING-LEFT: 20px! important; =
BACKGROUND-REPEAT: no-repeat; ZOOM: 1
}
.x-window-tl .x-panel-icon {
	BACKGROUND-POSITION: 0px 4px; PADDING-LEFT: 20px! important; =
BACKGROUND-REPEAT: no-repeat; ZOOM: 1
}
.x-panel-inline-icon {
	BACKGROUND-POSITION: 0px 0px; MARGIN-TOP: -1px; MARGIN-BOTTOM: -1px; =
VERTICAL-ALIGN: middle; WIDTH: 16px; MARGIN-RIGHT: 4px; =
BACKGROUND-REPEAT: no-repeat; HEIGHT: 16px
}
.x-panel-tc {
	BACKGROUND: url(../images/default/panel/top-bottom.gif) repeat-x 0px =
0px; OVERFLOW: hidden
}
.ext-strict .ext-ie7 .x-panel-tc {
	OVERFLOW: visible
}
.x-panel-tl {
	PADDING-LEFT: 6px; BACKGROUND: =
url(../images/default/panel/corners-sprite.gif) no-repeat 0px 0px; =
BORDER-BOTTOM: #99bbe8 1px solid; ZOOM: 1
}
.x-panel-tr {
	PADDING-RIGHT: 6px; BACKGROUND: =
url(../images/default/panel/corners-sprite.gif) no-repeat right 0px; =
ZOOM: 1
}
.x-panel-bc {
	BACKGROUND: url(../images/default/panel/top-bottom.gif) repeat-x 0px =
bottom; ZOOM: 1
}
.x-panel-bc .x-panel-footer {
	ZOOM: 1
}
.x-panel-bl {
	PADDING-LEFT: 6px; BACKGROUND: =
url(../images/default/panel/corners-sprite.gif) no-repeat 0px bottom; =
ZOOM: 1
}
.x-panel-br {
	PADDING-RIGHT: 6px; BACKGROUND: =
url(../images/default/panel/corners-sprite.gif) no-repeat right bottom; =
ZOOM: 1
}
.x-panel-mc {
	BORDER-RIGHT: 0px; PADDING-RIGHT: 0px; BORDER-TOP: 0px; PADDING-LEFT: =
0px; BACKGROUND: #dfe8f6; PADDING-BOTTOM: 0px; MARGIN: 0px; FONT: 11px =
tahoma,arial,helvetica,sans-serif; BORDER-LEFT: 0px; PADDING-TOP: 6px; =
BORDER-BOTTOM: 0px
}
.x-panel-mc .x-panel-body {
	BORDER-RIGHT: 0px; BORDER-TOP: 0px; BACKGROUND: none transparent scroll =
repeat 0% 0%; BORDER-LEFT: 0px; BORDER-BOTTOM: 0px
}
.x-panel-ml {
	PADDING-LEFT: 6px; BACKGROUND: =
url(../images/default/panel/left-right.gif) #fff repeat-y 0px 0px; ZOOM: =
1
}
.x-panel-mr {
	PADDING-RIGHT: 6px; BACKGROUND: =
url(../images/default/panel/left-right.gif) repeat-y right 0px; ZOOM: 1
}
.x-panel-bc .x-panel-footer {
	PADDING-BOTTOM: 6px
}
.x-panel-nofooter .x-panel-bc {
	FONT-SIZE: 0px; LINE-HEIGHT: 0; HEIGHT: 6px
}
.x-panel-nofooter .x-window-bc {
	FONT-SIZE: 0px; LINE-HEIGHT: 0; HEIGHT: 6px
}
.x-panel-bwrap {
	LEFT: 0px; OVERFLOW: hidden; ZOOM: 1; TOP: 0px
}
.x-panel-body {
	OVERFLOW: hidden; ZOOM: 1
}
.x-panel-collapsed .x-resizable-handle {
	DISPLAY: none
}
.ext-gecko .x-panel-animated DIV {
	OVERFLOW: hidden! important
}
.x-plain-body {
	OVERFLOW: hidden
}
.x-plain-bbar .x-toolbar {
	PADDING-RIGHT: 2px; PADDING-LEFT: 2px; PADDING-BOTTOM: 2px; OVERFLOW: =
hidden; PADDING-TOP: 2px
}
.x-plain-tbar .x-toolbar {
	PADDING-RIGHT: 2px; PADDING-LEFT: 2px; PADDING-BOTTOM: 2px; OVERFLOW: =
hidden; PADDING-TOP: 2px
}
.x-plain-bwrap {
	OVERFLOW: hidden; ZOOM: 1
}
.x-plain {
	OVERFLOW: hidden
}
.x-tool {
	BACKGROUND: url(../images/default/panel/tool-sprites.gif) no-repeat; =
FLOAT: right; MARGIN-LEFT: 2px; OVERFLOW: hidden; WIDTH: 15px; CURSOR: =
pointer; HEIGHT: 15px
}
.x-tool-toggle {
	BACKGROUND-POSITION: 0px -60px
}
.x-tool-toggle-over {
	BACKGROUND-POSITION: -15px -60px
}
.x-panel-collapsed .x-tool-toggle {
	BACKGROUND-POSITION: 0px -75px
}
.x-panel-collapsed .x-tool-toggle-over {
	BACKGROUND-POSITION: -15px -75px
}
.x-tool-close {
	BACKGROUND-POSITION: 0px 0px
}
.x-tool-close-over {
	BACKGROUND-POSITION: -15px 0px
}
.x-tool-minimize {
	BACKGROUND-POSITION: 0px -15px
}
.x-tool-minimize-over {
	BACKGROUND-POSITION: -15px -15px
}
.x-tool-maximize {
	BACKGROUND-POSITION: 0px -30px
}
.x-tool-maximize-over {
	BACKGROUND-POSITION: -15px -30px
}
.x-tool-restore {
	BACKGROUND-POSITION: 0px -45px
}
.x-tool-restore-over {
	BACKGROUND-POSITION: -15px -45px
}
.x-tool-gear {
	BACKGROUND-POSITION: 0px -90px
}
.x-tool-gear-over {
	BACKGROUND-POSITION: -15px -90px
}
.x-tool-pin {
	BACKGROUND-POSITION: 0px -135px
}
.x-tool-pin-over {
	BACKGROUND-POSITION: -15px -135px
}
.x-tool-unpin {
	BACKGROUND-POSITION: 0px -150px
}
.x-tool-unpin-over {
	BACKGROUND-POSITION: -15px -150px
}
.x-tool-right {
	BACKGROUND-POSITION: 0px -165px
}
.x-tool-right-over {
	BACKGROUND-POSITION: -15px -165px
}
.x-tool-left {
	BACKGROUND-POSITION: 0px -180px
}
.x-tool-left-over {
	BACKGROUND-POSITION: -15px -180px
}
.x-tool-up {
	BACKGROUND-POSITION: 0px -210px
}
.x-tool-up-over {
	BACKGROUND-POSITION: -15px -210px
}
.x-tool-down {
	BACKGROUND-POSITION: 0px -195px
}
.x-tool-down-over {
	BACKGROUND-POSITION: -15px -195px
}
.x-tool-refresh {
	BACKGROUND-POSITION: 0px -225px
}
.x-tool-refresh-over {
	BACKGROUND-POSITION: -15px -225px
}
.x-tool-minus {
	BACKGROUND-POSITION: 0px -255px
}
.x-tool-minus-over {
	BACKGROUND-POSITION: -15px -255px
}
.x-tool-plus {
	BACKGROUND-POSITION: 0px -240px
}
.x-tool-plus-over {
	BACKGROUND-POSITION: -15px -240px
}
.x-tool-search {
	BACKGROUND-POSITION: 0px -270px
}
.x-tool-search-over {
	BACKGROUND-POSITION: -15px -270px
}
.x-tool-save {
	BACKGROUND-POSITION: 0px -285px
}
.x-tool-save-over {
	BACKGROUND-POSITION: -15px -285px
}
.x-tool-help {
	BACKGROUND-POSITION: 0px -300px
}
.x-tool-help-over {
	BACKGROUND-POSITION: -15px -300px
}
.x-tool-print {
	BACKGROUND-POSITION: 0px -315px
}
.x-tool-print-over {
	BACKGROUND-POSITION: -15px -315px
}
.x-panel-ghost {
	Z-INDEX: 12000; BACKGROUND: #cbddf3; FILTER: alpha(opacity=3D65); LEFT: =
0px; OVERFLOW: hidden; POSITION: absolute; TOP: 0px; opacity: .65; =
-moz-opacity: .65
}
.x-panel-ghost UL {
	BORDER-RIGHT: #99bbe8 1px solid; PADDING-RIGHT: 0px; BORDER-TOP: =
#99bbe8 0px; DISPLAY: block; PADDING-LEFT: 0px; FONT-SIZE: 0px; =
PADDING-BOTTOM: 0px; MARGIN: 0px; OVERFLOW: hidden; BORDER-LEFT: #99bbe8 =
1px solid; LINE-HEIGHT: 0; PADDING-TOP: 0px; BORDER-BOTTOM: #99bbe8 1px =
solid
}
.x-panel-ghost * {
	CURSOR: move! important
}
.x-panel-dd-spacer {
	BORDER-RIGHT: #99bbe8 2px dashed; BORDER-TOP: #99bbe8 2px dashed; =
BORDER-LEFT: #99bbe8 2px dashed; BORDER-BOTTOM: #99bbe8 2px dashed
}
.x-panel-btns-ct {
	PADDING-RIGHT: 5px; PADDING-LEFT: 5px; PADDING-BOTTOM: 5px; =
PADDING-TOP: 5px
}
.x-panel-btns-ct .x-btn {
	CLEAR: none; FLOAT: right
}
.x-panel-btns-ct .x-panel-btns TD {
	BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; =
BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; =
PADDING-TOP: 0px; BORDER-RIGHT-WIDTH: 0px
}
.x-panel-btns-ct .x-panel-btns-right TABLE {
	CLEAR: none; FLOAT: right
}
.x-panel-btns-ct .x-panel-btns-left TABLE {
	CLEAR: none; FLOAT: left
}
.x-panel-btns-ct .x-panel-btns-center {
	TEXT-ALIGN: center
}
.x-panel-btns-ct .x-panel-btns-center TABLE {
	MARGIN: 0px auto
}
.x-panel-btns-ct TABLE TD.x-panel-btn-td {
	PADDING-RIGHT: 3px; PADDING-LEFT: 3px; PADDING-BOTTOM: 3px; =
PADDING-TOP: 3px
}
.x-panel-btns-ct .x-btn-focus .x-btn-left {
	BACKGROUND-POSITION: 0px -147px
}
.x-panel-btns-ct .x-btn-focus .x-btn-right {
	BACKGROUND-POSITION: 0px -168px
}
.x-panel-btns-ct .x-btn-focus .x-btn-center {
	BACKGROUND-POSITION: 0px -189px
}
.x-panel-btns-ct .x-btn-over .x-btn-left {
	BACKGROUND-POSITION: 0px -63px
}
.x-panel-btns-ct .x-btn-over .x-btn-right {
	BACKGROUND-POSITION: 0px -84px
}
.x-panel-btns-ct .x-btn-over .x-btn-center {
	BACKGROUND-POSITION: 0px -105px
}
.x-panel-btns-ct .x-btn-click .x-btn-center {
	BACKGROUND-POSITION: 0px -126px
}
.x-panel-btns-ct .x-btn-click .x-btn-right {
	BACKGROUND-POSITION: 0px -84px
}
.x-panel-btns-ct .x-btn-click .x-btn-left {
	BACKGROUND-POSITION: 0px -63px
}

------=_NextPart_000_0000_01CAD35F.003A3910
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.ncbi.nlm.nih.gov/core/ext.ncbi/1/css/themes/xtheme-oldentrez.css

.x-panel {
	BORDER-LEFT-COLOR: #d0d0d0; BORDER-BOTTOM-COLOR: #d0d0d0; =
BORDER-TOP-STYLE: solid; BORDER-TOP-COLOR: #d0d0d0; BORDER-RIGHT-STYLE: =
solid; BORDER-LEFT-STYLE: solid; BORDER-RIGHT-COLOR: #d0d0d0; =
BORDER-BOTTOM-STYLE: solid
}
.x-panel-header {
	BORDER-RIGHT: #d0d0d0 1px solid; BORDER-TOP: #d0d0d0 1px solid; =
BACKGROUND-IMAGE: url(../../img/oldentrez/panel/white-top-bottom.gif); =
BORDER-LEFT: #d0d0d0 1px solid; COLOR: #333; BORDER-BOTTOM: #d0d0d0 1px =
solid
}
.x-panel-body {
	BORDER-LEFT-COLOR: #d0d0d0; BORDER-BOTTOM-COLOR: #d0d0d0; =
BORDER-TOP-COLOR: #d0d0d0; BORDER-RIGHT-COLOR: #d0d0d0
}
.x-panel-bbar .x-toolbar {
	BORDER-LEFT-COLOR: #d0d0d0; BORDER-BOTTOM-COLOR: #d0d0d0; =
BORDER-TOP-COLOR: #d0d0d0; BORDER-RIGHT-COLOR: #d0d0d0
}
.x-panel-tbar .x-toolbar {
	BORDER-LEFT-COLOR: #d0d0d0; BORDER-BOTTOM-COLOR: #d0d0d0; =
BORDER-TOP-COLOR: #d0d0d0; BORDER-RIGHT-COLOR: #d0d0d0
}
.x-panel-tbar-noheader .x-toolbar {
	BORDER-LEFT-COLOR: #d0d0d0; BORDER-BOTTOM-COLOR: #d0d0d0; =
BORDER-TOP-COLOR: #d0d0d0; BORDER-RIGHT-COLOR: #d0d0d0
}
.x-panel-mc .x-panel-tbar .x-toolbar {
	BORDER-LEFT-COLOR: #d0d0d0; BORDER-BOTTOM-COLOR: #d0d0d0; =
BORDER-TOP-COLOR: #d0d0d0; BORDER-RIGHT-COLOR: #d0d0d0
}
.x-panel-body-noheader {
	BORDER-LEFT-COLOR: #d0d0d0; BORDER-BOTTOM-COLOR: #d0d0d0; =
BORDER-TOP-COLOR: #d0d0d0; BORDER-RIGHT-COLOR: #d0d0d0
}
.x-panel-mc .x-panel-body {
	BORDER-LEFT-COLOR: #d0d0d0; BORDER-BOTTOM-COLOR: #d0d0d0; =
BORDER-TOP-COLOR: #d0d0d0; BORDER-RIGHT-COLOR: #d0d0d0
}
.x-panel-tl .x-panel-header {
	COLOR: #333
}
.x-panel-tc {
	BACKGROUND-IMAGE: url(../../img/oldentrez/panel/top-bottom.gif)
}
.x-panel-tl {
	BORDER-LEFT-COLOR: #d0d0d0; BACKGROUND-IMAGE: =
url(../../img/oldentrez/panel/corners-sprite.gif); BORDER-BOTTOM-COLOR: =
#d0d0d0; BORDER-TOP-COLOR: #d0d0d0; BORDER-RIGHT-COLOR: #d0d0d0
}
.x-panel-tr {
	BACKGROUND-IMAGE: url(../../img/oldentrez/panel/corners-sprite.gif)
}
.x-panel-bc {
	BACKGROUND-IMAGE: url(../../img/oldentrez/panel/top-bottom.gif)
}
.x-panel-bl {
	BACKGROUND-IMAGE: url(../../img/oldentrez/panel/corners-sprite.gif)
}
.x-panel-br {
	BACKGROUND-IMAGE: url(../../img/oldentrez/panel/corners-sprite.gif)
}
.x-panel-mc {
	BACKGROUND: #f1f1f1
}
.x-panel-mc .x-panel-body {
	BORDER-RIGHT: 0px; BORDER-TOP: 0px; BACKGROUND: none transparent scroll =
repeat 0% 0%; BORDER-LEFT: 0px; BORDER-BOTTOM: 0px
}
.x-panel-ml {
	BACKGROUND-IMAGE: url(../../img/oldentrez/panel/left-right.gif)
}
.x-panel-mr {
	BACKGROUND-IMAGE: url(../../img/oldentrez/panel/left-right.gif)
}
.x-tool {
	BACKGROUND-IMAGE: url(../../img/oldentrez/panel/tool-sprites.gif)
}
.x-panel-ghost {
	BACKGROUND: #e0e0e0
}
.x-panel-ghost UL {
	BORDER-LEFT-COLOR: #b0b0b0; BORDER-BOTTOM-COLOR: #b0b0b0; =
BORDER-TOP-COLOR: #b0b0b0; BORDER-RIGHT-COLOR: #b0b0b0
}
.x-grid-panel .x-panel-mc .x-panel-body {
	BORDER-RIGHT: #d0d0d0 1px solid; BORDER-TOP: #d0d0d0 1px solid; =
BORDER-LEFT: #d0d0d0 1px solid; BORDER-BOTTOM: #d0d0d0 1px solid
}
.x-btn-left {
	BACKGROUND-IMAGE: url(../../img/oldentrez/button/btn-sprite.gif)
}
.x-btn-right {
	BACKGROUND-IMAGE: url(../../img/oldentrez/button/btn-sprite.gif)
}
.x-btn-center {
	BACKGROUND-IMAGE: url(../../img/oldentrez/button/btn-sprite.gif)
}
.x-border-layout-ct {
	BACKGROUND: #f0f0f0
}
.x-accordion-hd {
	BACKGROUND-IMAGE: url(../../img/oldentrez/panel/light-hd.gif)
}
.x-layout-collapsed {
	BORDER-LEFT-COLOR: #e0e0e0; BORDER-BOTTOM-COLOR: #e0e0e0; =
BORDER-TOP-COLOR: #e0e0e0; BACKGROUND-COLOR: #eee; BORDER-RIGHT-COLOR: =
#e0e0e0
}
.x-layout-collapsed-over {
	BACKGROUND-COLOR: #fbfbfb
}
.x-tip .x-tip-top {
	BACKGROUND-IMAGE: url(../../img/oldentrez/qtip/tip-sprite.gif)
}
.x-tip .x-tip-top-left {
	BACKGROUND-IMAGE: url(../../img/oldentrez/qtip/tip-sprite.gif)
}
.x-tip .x-tip-top-right {
	BACKGROUND-IMAGE: url(../../img/oldentrez/qtip/tip-sprite.gif)
}
.x-tip .x-tip-ft {
	BACKGROUND-IMAGE: url(../../img/oldentrez/qtip/tip-sprite.gif)
}
.x-tip .x-tip-ft-left {
	BACKGROUND-IMAGE: url(../../img/oldentrez/qtip/tip-sprite.gif)
}
.x-tip .x-tip-ft-right {
	BACKGROUND-IMAGE: url(../../img/oldentrez/qtip/tip-sprite.gif)
}
.x-tip .x-tip-bd-left {
	BACKGROUND-IMAGE: url(../../img/oldentrez/qtip/tip-sprite.gif)
}
.x-tip .x-tip-bd-right {
	BACKGROUND-IMAGE: url(../../img/oldentrez/qtip/tip-sprite.gif)
}
.x-toolbar {
	BORDER-LEFT-COLOR: #d0d0d0; BACKGROUND: =
url(../../img/oldentrez/toolbar/bg.gif) #f0f4f5 repeat-x left top; =
BORDER-BOTTOM-COLOR: #d0d0d0; BORDER-TOP-COLOR: #d0d0d0; =
BORDER-RIGHT-COLOR: #d0d0d0
}
.x-toolbar BUTTON {
	COLOR: #444
}
.x-toolbar .x-btn-menu-arrow-wrap .x-btn-center BUTTON {
	BACKGROUND-IMAGE: url(../../img/oldentrez/toolbar/btn-arrow.gif)
}
.x-toolbar .x-btn-text-icon .x-btn-menu-arrow-wrap .x-btn-center BUTTON =
{
	BACKGROUND-IMAGE: url(../../img/oldentrez/toolbar/btn-arrow.gif)
}
.x-toolbar .x-btn-over .x-btn-left {
	BACKGROUND-IMAGE: url(../../img/oldentrez/toolbar/tb-btn-sprite.gif)
}
.x-toolbar .x-btn-over .x-btn-right {
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------=_NextPart_000_0000_01CAD35F.003A3910
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BODY UL.jig-ncbimenu A:focus {
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------=_NextPart_000_0000_01CAD35F.003A3910
Content-Type: text/css;
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.port H2.x-panel-header-text {
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.port H3.x-panel-header-text {
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.port H4.x-panel-header-text {
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.port H5.x-panel-header-text {
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.port .x-panel-header {
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}
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}
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}
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}
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}
.x-panel-mc .x-panel-body {
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}
.x-panel-tl .x-panel-header {
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}
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}
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BORDER-BOTTOM-COLOR: #d0d0d0; BORDER-TOP-COLOR: #d0d0d0; =
BORDER-RIGHT-COLOR: #d0d0d0
}
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}
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}
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}
.x-panel-br {
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}
.x-panel-mc {
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}
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DIV#HTDisplay DIV.action {
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}
DIV#HTDisplay DIV.action A {
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DIV#HTDisplay P {
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}
DIV#HTDisplay UL LI .thumb {
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}
DIV#HTDisplay P {
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}
DIV#HTDisplay .action A.HTOff {
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}
DIV.HTOff#HTDisplay .action A.HTOn {
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DIV.HTOff#HTDisplay P.HTOff {
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DIV.HTOn#HTDisplay .action A.HTOn {
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}
DIV#HTDisplay UL.hide {
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}
IMG.htbpoparrow {
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}
DIV#htbpopper {
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FONT-FAMILY: Verdana; POSITION: absolute; BACKGROUND-COLOR: #f0f8ff; =
opacity: 100
}
DIV.popper P {
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}
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P#htbpcit {
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}
DIV#HTDisplay UL LI SPAN.thumb {
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}
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}
#content LI.DiscoveryItem A {
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UNKNOWN {
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}
#content LI.DiscoveryAllLinks {
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}
#content LI.DiscoveryAllLinks A {
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}
DIV.popper EM.cit {
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}
DIV.popper EM.author {
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}
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FONT-WEIGHT: 500; FONT-SIZE: 0.9em; FLOAT: left; PADDING-BOTTOM: 0px; =
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}
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DIV.popper SPAN.reviewflag {
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}
BODY #col1 {
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#00c 1px solid; BORDER-BOTTOM: #00c 1px solid
}
BODY #col2 {
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}
BODY #subheader {
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#00c 1px solid; BORDER-BOTTOM: #00c 1px solid
}
BODY #subfooter {
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}
BODY #subfooter {
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}
#Table2 {
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}
TD.pmc-watermark {
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}
TD.format-menu {
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}
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PADDING-BOTTOM: 0px; MARGIN: 0px 0px 0px 0.2em; PADDING-TOP: 0px
}
TD.format-menu H2 {
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TD.format-menu UL LI {
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}
TD.format-menu UL LI A {
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}
TD.format-menu UL LI.selected {
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}
TD.content-cell > DIV TD.content-cell > DIV > DIV {
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}
TD.sidebar-cell {
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}
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}
DIV.cited-ref {
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POSITION: absolute
}
DIV.section-content > DIV.p {
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}
IMG.equation {
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DIV.ArticleRefDocsums {
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}
DIV.pmlinks UL.links LI.ovfl {
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DIV.cited-ref LI.ovfl {
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}
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}
DIV.cited-ref LI.ovfl SPAN.flag {
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P.desc {
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DIV.cited-ref UL {
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}
DIV.cited-ref A.all-articles {
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}
DIV.cited-ref A.all-articles:hover {
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}
DIV.port {
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DIV.x-panel-body {
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}
DIV.sameauthor-seemore {
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}
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}
SPAN.review {
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}

@media Print   =20
{
HTML BODY.pmc-body {
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}
TD.sidebar-cell DIV.port {
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}
#ArticleRefDocsums {
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}
.format-menu {
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}
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}
TD.content-cell {
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}
DIV.section-content > DIV.p {
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}
DIV.sec {
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}
#Table3 TD.sidebar-cell {
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}
TD.content-cell {
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}
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}
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}
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0px; LINE-HEIGHT: 1; PADDING-TOP: 0px; LIST-STYLE-TYPE: none
}
.mb A {
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underline; outline: none
}
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}
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}
UL.mb LI SPAN.delta {
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}
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}
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}
UL.mb LI.submenu-item UL {
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BORDER-LEFT: red 1px solid; BORDER-BOTTOM: red 1px solid; POSITION: =
absolute
}
UL.mb LI.active-item UL {
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}
UL.mb LI UL.opened {
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PADDING-BOTTOM: 2px; BORDER-LEFT: #ffffff 1px solid; PADDING-TOP: 2px; =
BORDER-BOTTOM: #ffffff 1px solid; BACKGROUND-COLOR: #ffffff
}
UL.mb LI UL.opened LI {
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}
UL.mb LI UL.opened LI.active-item {
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PADDING-BOTTOM: 0px; PADDING-TOP: 0px
}
UL.opened LI.active-item A {
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}
UL.mb LI UL A {
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3px; COLOR: #222222; PADDING-TOP: 2px; TEXT-DECORATION: none
}
TD.content-cell {
	MIN-WIDTH: 550px; WIDTH: 550px
}
#Table3 {
	TABLE-LAYOUT: fixed
}
.col-watermark {
	MIN-WIDTH: 20px; WIDTH: 20px
}
.col-content {
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}
.col-sidebar {
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------=_NextPart_000_0000_01CAD35F.003A3910
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.ncbi.nlm.nih.gov/corehtml/pmc/js/jquery.hoverIntent.min.js

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------=_NextPart_000_0000_01CAD35F.003A3910
Content-Type: application/octet-stream
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'x0017a':'z',=0A=
'x0017b':'Z',=0A=
'x0017c':'z',=0A=
'x0017d':'Z',=0A=
'x0017e':'z',=0A=
'x001f5':'g',=0A=
'x00405':'S',=0A=
'x00406':'I',=0A=
'x00408':'J',=0A=
'x00455':'s',=0A=
'x00456':'i',=0A=
'x00458':'j',=0A=
'x02102':'C',=0A=
'x0210b':'H',=0A=
'x0210c':'H',=0A=
'x02110':'I',=0A=
'x02111':'I',=0A=
'x02112':'L',=0A=
'x02115':'N',=0A=
'x02119':'P',=0A=
'x0211a':'Q',=0A=
'x0211c':'R',=0A=
'x0211d':'R',=0A=
'x0211e':'Rx',=0A=
'x02124':'Z',=0A=
'x0212b':'A',=0A=
'x0212c':'B',=0A=
'x0212f':'e',=0A=
'x02130':'E',=0A=
'x02131':'F',=0A=
'x02133':'M',=0A=
'x02134':'o',=0A=
'x0e05d':'h',=0A=
'x0e05f':'k',=0A=
'x0e3b2':'fj',=0A=
'x0e500':'A',=0A=
'x0e501':'B',=0A=
'x0e503':'D',=0A=
'x0e504':'E',=0A=
'x0e505':'F',=0A=
'x0e506':'G',=0A=
'x0e507':'H',=0A=
'x0e508':'I',=0A=
'x0e509':'J',=0A=
'x0e50a':'K',=0A=
'x0e50b':'L',=0A=
'x0e50c':'M',=0A=
'x0e50e':'O',=0A=
'x0e512':'S',=0A=
'x0e513':'T',=0A=
'x0e514':'U',=0A=
'x0e515':'V',=0A=
'x0e516':'W',=0A=
'x0e517':'X',=0A=
'x0e518':'Y',=0A=
'x0e520':'A',=0A=
'x0e522':'C',=0A=
'x0e523':'D',=0A=
'x0e526':'G',=0A=
'x0e529':'J',=0A=
'x0e52a':'K',=0A=
'x0e52d':'N',=0A=
'x0e52e':'O',=0A=
'x0e52f':'P',=0A=
'x0e530':'Q',=0A=
'x0e531':'R',=0A=
'x0e532':'S',=0A=
'x0e533':'T',=0A=
'x0e534':'U',=0A=
'x0e535':'V',=0A=
'x0e536':'W',=0A=
'x0e537':'X',=0A=
'x0e538':'Y',=0A=
'x0e539':'Z',=0A=
'x0e540':'a',=0A=
'x0e541':'b',=0A=
'x0e542':'c',=0A=
'x0e543':'d',=0A=
'x0e545':'f',=0A=
'x0e546':'g',=0A=
'x0e547':'h',=0A=
'x0e548':'i',=0A=
'x0e549':'j',=0A=
'x0e54a':'k',=0A=
'x0e54b':'l',=0A=
'x0e54c':'m',=0A=
'x0e54d':'n',=0A=
'x0e54f':'p',=0A=
'x0e550':'q',=0A=
'x0e551':'r',=0A=
'x0e552':'s',=0A=
'x0e553':'t',=0A=
'x0e554':'u',=0A=
'x0e555':'v',=0A=
'x0e556':'w',=0A=
'x0e557':'x',=0A=
'x0e558':'y',=0A=
'x0e559':'z',=0A=
'x0e560':'A',=0A=
'x0e561':'B',=0A=
'x0e562':'C',=0A=
'x0e563':'D',=0A=
'x0e564':'E',=0A=
'x0e565':'F',=0A=
'x0e566':'G',=0A=
'x0e569':'J',=0A=
'x0e56a':'K',=0A=
'x0e56b':'L',=0A=
'x0e56c':'M',=0A=
'x0e56d':'N',=0A=
'x0e56e':'O',=0A=
'x0e56f':'P',=0A=
'x0e570':'Q',=0A=
'x0e572':'S',=0A=
'x0e573':'T',=0A=
'x0e574':'U',=0A=
'x0e575':'V',=0A=
'x0e576':'W',=0A=
'x0e577':'X',=0A=
'x0e578':'Y',=0A=
'x0e579':'Z',=0A=
'x0e580':'a',=0A=
'x0e581':'b',=0A=
'x0e582':'c',=0A=
'x0e583':'d',=0A=
'x0e584':'e',=0A=
'x0e585':'f',=0A=
'x0e586':'g',=0A=
'x0e587':'h',=0A=
'x0e588':'i',=0A=
'x0e589':'j',=0A=
'x0e58a':'k',=0A=
'x0e58b':'l',=0A=
'x0e58c':'m',=0A=
'x0e58d':'n',=0A=
'x0e58e':'o',=0A=
'x0e58f':'p',=0A=
'x0e590':'q',=0A=
'x0e591':'r',=0A=
'x0e592':'s',=0A=
'x0e593':'t',=0A=
'x0e594':'u',=0A=
'x0e595':'v',=0A=
'x0e596':'w',=0A=
'x0e597':'x',=0A=
'x0e598':'y',=0A=
'x0e599':'z',=0A=
'x0ea00':'1',=0A=
'x0ea01':'1',=0A=
'x0ea02':'2',=0A=
'x0ea03':'2',=0A=
'x0ea04':'3',=0A=
'x0ea05':'3',=0A=
'x0ea06':'4',=0A=
'x0ea07':'a',=0A=
'x0ea08':'a',=0A=
'x0ea09':'A',=0A=
'x0ea0a':'a',=0A=
'x0ea0b':'A',=0A=
'x0ea0c':'AB',=0A=
'x0ea10':'b',=0A=
'x0ea11':'B',=0A=
'x0ea12':'b',=0A=
'x0ea13':'B',=0A=
'x0ea14':'B',=0A=
'x0ea15':'B',=0A=
'x0ea16':'BC',=0A=
'x0ea19':'c',=0A=
'x0ea1a':'c',=0A=
'x0ea1b':'c',=0A=
'x0ea1c':'C',=0A=
'x0ea1d':'c',=0A=
'x0ea1e':'c',=0A=
'x0ea1f':'c',=0A=
'x0ea20':'c',=0A=
'x0ea21':'c',=0A=
'x0ea25':'d',=0A=
'x0ea26':'D',=0A=
'x0ea27':'d',=0A=
'x0ea28':'D',=0A=
'x0ea29':'d',=0A=
'x0ea2a':'D',=0A=
'x0ea2b':'D',=0A=
'x0ea2f':'dl',=0A=
'x0ea30':'e',=0A=
'x0ea31':'e',=0A=
'x0ea32':'l',=0A=
'x0ea33':'l',=0A=
'x0ea34':'f',=0A=
'x0ea35':'f',=0A=
'x0ea36':'F',=0A=
'x0ea37':'f',=0A=
'x0ea38':'F',=0A=
'x0ea39':'f',=0A=
'x0ea3a':'F',=0A=
'x0ea3d':'g',=0A=
'x0ea3e':'g',=0A=
'x0ea3f':'G',=0A=
'x0ea40':'g',=0A=
'x0ea41':'G',=0A=
'x0ea46':'H',=0A=
'x0ea47':'h',=0A=
'x0ea48':'h',=0A=
'x0ea49':'i',=0A=
'x0ea4a':'I',=0A=
'x0ea4b':'j',=0A=
'x0ea4c':'J',=0A=
'x0ea4d':'J',=0A=
'x0ea4e':'J',=0A=
'x0ea4f':'k',=0A=
'x0ea50':'k',=0A=
'x0ea51':'k',=0A=
'x0ea52':'K',=0A=
'x0ea53':'K',=0A=
'x0ea54':'k',=0A=
'x0ea55':'l',=0A=
'x0ea56':'L',=0A=
'x0ea57':'l',=0A=
'x0ea5b':'lnV',=0A=
'x0ea5c':'m',=0A=
'x0ea5d':'M',=0A=
'x0ea5e':'m',=0A=
'x0ea5f':'m',=0A=
'x0ea63':'n',=0A=
'x0ea64':'n',=0A=
'x0ea65':'N',=0A=
'x0ea66':'n',=0A=
'x0ea67':'N',=0A=
'x0ea68':'n',=0A=
'x0ea69':'n',=0A=
'x0ea6a':'N',=0A=
'x0ea6d':'nv',=0A=
'x0ea6e':'O',=0A=
'x0ea6f':'O',=0A=
'x0ea70':'O',=0A=
'x0ea72':'F',=0A=
'x0ea73':'p',=0A=
'x0ea74':'P',=0A=
'x0ea75':'p',=0A=
'x0ea76':'P',=0A=
'x0ea77':'p',=0A=
'x0ea78':'P',=0A=
'x0ea79':'p',=0A=
'x0ea7a':'P',=0A=
'x0ea8d':'q',=0A=
'x0ea8e':'Q',=0A=
'x0ea8f':'q',=0A=
'x0ea90':'q',=0A=
'x0ea91':'Q',=0A=
'x0ea92':'q',=0A=
'x0ea93':'Q',=0A=
'x0ea94':'q',=0A=
'x0ea95':'q',=0A=
'x0ea96':'q',=0A=
'x0ea97':'Q',=0A=
'x0ea98':'r',=0A=
'x0ea99':'R',=0A=
'x0ea9a':'R',=0A=
'x0ea9b':'r',=0A=
'x0ea9c':'R',=0A=
'x0ea9d':'R',=0A=
'x0ea9e':'r',=0A=
'x0ea9f':'R',=0A=
'x0eaa0':'r',=0A=
'x0eaa1':'ri',=0A=
'x0eaa2':'R',=0A=
'x0eaa3':'r',=0A=
'x0eaa4':'r1',=0A=
'x0eaa5':'RE',=0A=
'x0eaa9':'ri',=0A=
'x0eaaa':'rj',=0A=
'x0eaab':'rN',=0A=
'x0eaac':'s',=0A=
'x0eaad':'S',=0A=
'x0eaae':'S',=0A=
'x0eaaf':'s',=0A=
'x0eab0':'S',=0A=
'x0eab1':'s',=0A=
'x0eab2':'S',=0A=
'x0eab3':'S',=0A=
'x0eab4':'B',=0A=
'x0eab5':'E',=0A=
'x0eab6':'G',=0A=
'x0eab7':'P',=0A=
'x0eab8':'Q',=0A=
'x0eab9':'t',=0A=
'x0eaba':'T',=0A=
'x0eabb':'T',=0A=
'x0eabc':'t',=0A=
'x0eabd':'T',=0A=
'x0eabe':'t',=0A=
'x0eabf':'T',=0A=
'x0eac5':'TT',=0A=
'x0eac6':'u',=0A=
'x0eac7':'u',=0A=
'x0eac8':'U',=0A=
'x0eac9':'u',=0A=
'x0eaca':'u',=0A=
'x0eacc':'V',=0A=
'x0eacd':'v',=0A=
'x0eace':'v',=0A=
'x0eacf':'V',=0A=
'x0ead0':'v',=0A=
'x0ead1':'V',=0A=
'x0ead2':'v',=0A=
'x0ead3':'v',=0A=
'x0ead4':'V',=0A=
'x0ead5':'v',=0A=
'x0eadc':'w',=0A=
'x0eadd':'w',=0A=
'x0eade':'w',=0A=
'x0eadf':'x',=0A=
'x0eae0':'X',=0A=
'x0eae1':'x',=0A=
'x0eae2':'X',=0A=
'x0eae3':'x',=0A=
'x0eae4':'x',=0A=
'x0eae5':'X',=0A=
'x0eae6':'x',=0A=
'x0eae8':'y',=0A=
'x0eae9':'Y',=0A=
'x0eaea':'y',=0A=
'x0eaeb':'Y',=0A=
'x0eaec':'y',=0A=
'x0eaed':'y',=0A=
'x0eaee':'z',=0A=
'x0eaef':'Z',=0A=
'x0eaf0':'z',=0A=
'x0eaf1':'z',=0A=
'x0eaf2':'Z',=0A=
'x0eaf3':'z',=0A=
'x0eaf4':'z',=0A=
'x0eafc':'B',=0A=
'x0eaff':'A',=0A=
'x0eb04':'w',=0A=
'x0eb05':'m',=0A=
'x0eb06':'M',=0A=
'x0eb07':'E',=0A=
'x0eb08':'W',=0A=
'x0eb0a':'h',=0A=
'x0eb0b':'n',=0A=
'x0eb0d':'H',=0A=
'x0eb0f':'M',=0A=
'x0eb10':'m',=0A=
'x0eb11':'Z',=0A=
'x0eb12':'g',=0A=
'x0eb14':'C',=0A=
'x0eb15':'E',=0A=
'x0eb19':'W',=0A=
'x0eb1a':'a',=0A=
'x0eb1b':'b',=0A=
'x0eb1c':'S',=0A=
'x0eb1e':'J',=0A=
'x0eb1f':'b',=0A=
'x0eb20':'Z',=0A=
'x0eb21':'L',=0A=
'x0eb22':'g',=0A=
'x0fb00':'ffi',=0A=
'x0fb01':'fi',=0A=
'x0fb02':'fl',=0A=
'x0fb03':'ffi',=0A=
'x0fb04':'ffl'=0A=
    }=0A=
=0A=
// Called from many places to handle links.=0A=
// Params:=0A=
//     link:         URL. 'this' to reuse same window, else ""=0A=
//     windowname:   Reference name for window=0A=
//     additional:   Attributes for window.open, from:=0A=
//        width, height, resizable, scrollbars, toolbar, location, =
directories,=0A=
//        status, menubar, copyhistory.=0A=
// Notes: Book version always provides menubar, toolbar, etc.=0A=
//=0A=
function startTargetBook(link,windowname,width,height,additional)=0A=
{=0A=
    startTarget(link,windowname,width,height,additional)=0A=
}=0A=
=0A=
=0A=
//=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=0A=
function startTarget(link,windowname,width,height, additional)=0A=
{=0A=
=0A=
    if(! window.focus)=0A=
        return;=0A=
=0A=
    var sizestring =3D ",width=3D" + width + ",height=3D" + height;=0A=
	var opt =3D =
"menubar=3Dno,toolbar=3Dno,status=3Dno,scrollbars=3Dyes,resizable=3Dyes,d=
ependent=3Dyes,location=3Dno";=0A=
    var allOptions =3D opt + sizestring;=0A=
    if (additional) { allOptions =3D allOptions + ',' + additional; }=0A=
=0A=
    windowname =3D String(windowname).replace(/-/g, "_")=0A=
    var moveToXDefault =3D 75=0A=
    var moveToYDefault =3D 50=0A=
=0A=
    var moveToX =3D moveToXDefault=0A=
    var moveToY =3D moveToYDefault=0A=
    var yAdj =3D 50=0A=
    var yAdjDelta =3D 25=0A=
=0A=
    var barNames =3D ['menubar', 'toolbar', 'statusbar']=0A=
    for (var idx in barNames)=0A=
    {=0A=
	if (String(allOptions).lastIndexOf(barNames[idx] + '=3Dyes') >=0A=
	    String(allOptions).lastIndexOf(barNames[idx] + '=3Dno'))=0A=
		yAdj +=3D yAdjDelta=0A=
    }=0A=
=0A=
    try {=0A=
        if (window.screen.width && width > 0)=0A=
            moveToX =3D Math.ceil((window.screen.width - width)) - 15=0A=
        moveToX =3D (moveToX > 0 ? moveToX : moveToXDefault)=0A=
=0A=
        if (window.screen.height && height > 0)=0A=
            moveToY =3D Math.ceil((window.screen.height - height)) - yAdj=0A=
        moveToY =3D (moveToY > 0 ? moveToY : moveToYDefault)=0A=
    }=0A=
    catch (e) {}=0A=
=0A=
    var wLeftTopCornerOptions =3D ',left=3D' + moveToX + ',top=3D' + =
moveToY + ',screenX=3D' + moveToX + ',screenY=3D' + moveToY=0A=
=0A=
    allOptions =3D allOptions + wLeftTopCornerOptions=0A=
=0A=
    w =3D window.open (link, windowname, allOptions)=0A=
    w.focus()=0A=
=0A=
    link.target=3Dwindowname=0A=
=0A=
    return false;=0A=
}=0A=
=0A=
=0A=
//=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=0A=
function focuswin(windowname)=0A=
{=0A=
    windowname =3D String(windowname).replace(/-/g, "_")=0A=
    w =3D =
window.open("",windowname,"menubar=3Dyes,scrollbars=3Dyes,toolbar=3Dyes,l=
ocation=3Dyes,status=3Dyes,directories=3Dyes,resizable=3Dyes");=0A=
    w.focus();=0A=
=0A=
    return true;=0A=
}=0A=
=0A=
=0A=
//=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=0A=
function reverseString (inStr)=0A=
{=0A=
    var outStr =3D ''=0A=
=0A=
    for (i =3D0; i <=3D inStr.length; i++)=0A=
    {=0A=
	outStr =3D inStr.charAt(i) + outStr=0A=
    }=0A=
=0A=
    return outStr=0A=
}=0A=
=0A=
=0A=
//=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=0A=
function reverseAndReplaceString (inStr, findStr, newStr)=0A=
{=0A=
    return reverseString(inStr).replace(findStr, newStr)=0A=
}=0A=
=0A=
//=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=0A=
function initUnObscureEmail (className, innerHTML)=0A=
{=0A=
=0A=
    try{=0A=
        if (window.addEventListener)=0A=
            window.addEventListener('load', function() {unObscureEmail =
(className, innerHTML)}, false)=0A=
        else if (window.attachEvent)=0A=
            window.attachEvent('onload', function() {unObscureEmail =
(className, innerHTML)})=0A=
    }catch(e){=0A=
    }=0A=
}=0A=
=0A=
//=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=0A=
function unObscureEmail (className, innerHTML)=0A=
{=0A=
    try {=0A=
        if (typeof(unObscuredEmails) =3D=3D 'undefined')=0A=
            unObscuredEmails =3D new Array()=0A=
=0A=
        if (! unObscuredEmails[className])=0A=
        {=0A=
=0A=
            var elmnts =3D document.getElementsByTagName("span")=0A=
            for (var i =3D 0, len =3D elmnts.length; i < len; i++)=0A=
            {=0A=
                if (elmnts.item(i).className.indexOf(className, " ") =
!=3D -1)=0A=
                {=0A=
                    elmnts.item(i).innerHTML     =3D innerHTML;=0A=
                    unObscuredEmails[className]  =3D true=0A=
=0A=
                }=0A=
            }=0A=
        }=0A=
    }=0A=
    catch(e){=0A=
    }=0A=
}=0A=
=0A=
//=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=0A=
function pubMedDbLinkSubmit(control) {=0A=
    try{=0A=
        if (control.pubmedOption)=0A=
            control =3D control.pubmedOption=0A=
        if (control.options)=0A=
            location.href =3D =
control.options[control.options.selectedIndex].value;=0A=
    }=0A=
    catch(e){}=0A=
    return false;=0A=
}=0A=
=0A=
//=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=0A=
// elements "a", "area" with "ref" attribute report to redirect utility =
before dereferencing link=0A=
//=0A=
function initRedirectClicks(report_url) {=0A=
=0A=
    if (report_url && typeof jQuery !=3D 'undefined')=0A=
        jQuery(document).click(function(e) {=0A=
=0A=
            //=0A=
            if (!e) return true;=0A=
            //=0A=
            if (e.shiftKey || e.altKey || e.ctrlKey) return true;=0A=
            //=0A=
            var target =3D jQuery(e.target).closest("a[ref], area[ref]");=0A=
            if (!target || target.length =3D=3D 0) return true;=0A=
            //=0A=
            var ref  =3D target.attr("ref");=0A=
            var href =3D target.attr("href")=0A=
=0A=
            // special processing of href element for pubmed's authors=0A=
            if (typeof (href) !=3D 'undefined' && =
href.indexOf('sites/entrez') && href.indexOf('%5Bauth%5D'))=0A=
            {=0A=
                var newHref =3D replacePubMedUnsupportedChars(href);=0A=
                if (newHref !=3D href)=0A=
                    target.attr("href", newHref)=0A=
            }=0A=
=0A=
            // append ref with value of href if 'redirect-to-url' is =
missing in ref.=0A=
            if (typeof (ref) !=3D 'undefined' && =
ref.indexOf('redirect-to-url') =3D=3D -1)=0A=
            {=0A=
                ref =3D ref + '&redirect-to-url=3D' + =
encodeURIComponent(href)=0A=
                //=0A=
                jQuery.ajax({=0A=
                   type:    "HEAD",=0A=
                   url:     report_url + '?' + ref,=0A=
                   timeout: 1000,=0A=
                   async:   false=0A=
                });=0A=
            }=0A=
=0A=
            return true;=0A=
        });=0A=
};=0A=
=0A=
// function to replace unsupported charachters in author names to be =
searchable in PubMed database.=0A=
function replacePubMedUnsupportedChars (str)=0A=
{=0A=
    // replace low 128 chars automatically=0A=
    str =3D str.replace(/%26%23(x000[0-7][0-9a-f])%3b/ig,=0A=
        function($0, $1){=0A=
            return (String.fromCharCode('0' + $1))=0A=
        }=0A=
    )=0A=
    // replace others based on map and if it is not in the map replace =
with empty string.=0A=
    str =3D =
str.replace(/%26%23(x[0-9a-f][0-9a-f][0-9a-f][0-9a-f][0-9a-f])%3b/ig,=0A=
        function($0, $1){=0A=
            return (pubmedUnsupportedCharMap[$1] ? =
pubmedUnsupportedCharMap[$1] : '')=0A=
        }=0A=
    )=0A=
    return str;=0A=
}=0A=
//=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=0A=
// Based on script written by Dean Edwards, 2005=0A=
// http://dean.edwards.name/=0A=
=0A=
var pmc_script =3D {=0A=
	addEvent: function(element, type, handler) {=0A=
		// assign each event handler a unique ID=0A=
		if (!handler.$$guid) handler.$$guid =3D this.addEvent_guid++;=0A=
		// create a hash table of event types for the element=0A=
		if (!element.events) element.events =3D {};=0A=
		// create a hash table of event handlers for each element/event pair=0A=
		var handlers =3D element.events[type];=0A=
		if (!handlers) {=0A=
			handlers =3D element.events[type] =3D {};=0A=
			// store the existing event handler (if there is one)=0A=
			if (element["on" + type]) {=0A=
				handlers[0] =3D element["on" + type];=0A=
			}=0A=
		}=0A=
		// store the event handler in the hash table=0A=
		handlers[handler.$$guid] =3D handler;=0A=
		// assign a global event handler to do all the work=0A=
		element["on" + type] =3D handleEvent;=0A=
=0A=
=0A=
		function handleEvent(event) {=0A=
			var returnValue =3D true;=0A=
			// grab the event object (IE uses a global event object)=0A=
			event =3D event || fixEvent(window.event);=0A=
			// get a reference to the hash table of event handlers=0A=
			var handlers =3D this.events[event.type];=0A=
			// execute each event handler=0A=
			for (var i in handlers) {=0A=
				this.$$handleEvent =3D handlers[i];=0A=
				if (this.$$handleEvent(event) =3D=3D=3D false) {=0A=
					returnValue =3D false;=0A=
				}=0A=
			}=0A=
			return returnValue;=0A=
		};=0A=
=0A=
		function fixEvent(event) {=0A=
			// add W3C standard event methods=0A=
			event.preventDefault =3D fixEvent.preventDefault;=0A=
			event.stopPropagation =3D fixEvent.stopPropagation;=0A=
			return event;=0A=
		};=0A=
=0A=
		fixEvent.preventDefault =3D function() {=0A=
			this.returnValue =3D false;=0A=
		};=0A=
=0A=
		fixEvent.stopPropagation =3D function() {=0A=
			this.cancelBubble =3D true;=0A=
		};=0A=
	},=0A=
=0A=
	// a counter used to create unique IDs=0A=
	addEvent_guid: 1,=0A=
=0A=
	removeEvent: function(element, type, handler) {=0A=
		// delete the event handler from the hash table=0A=
		if (element.events && element.events[type]) {=0A=
			delete element.events[type][handler.$$guid];=0A=
		}=0A=
	}=0A=
}=0A=
=0A=
=0A=
// =
*************************************************************************=
***=0A=
//  implementation of figure's popup using jQuery and =
jQuery.fn.hoverIntent lib=0A=
//  it set appropriate handlers for all "a" elements with "figpopup" =
class name.=0A=
// =
*************************************************************************=
***=0A=
if (typeof jQuery !=3D 'undefined' && typeof jQuery.fn.hoverIntent !=3D =
'undefined') {=0A=
    (function($j) {=0A=
        // =
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D  hoverHandlers =
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=3D=3D=0A=
        var hoverHandlers =3D function  (opts) {=0A=
            this.config =3D {}=0A=
            this.self =3D this;=0A=
            this.__init(opts)=0A=
        }=0A=
=0A=
         $j.fn.extend(hoverHandlers.prototype,  {=0A=
                //=0A=
                __init: function(opts) {=0A=
                    $j.fn.extend(this.config, opts)=0A=
                },=0A=
                //=0A=
                over: function (el) {=0A=
                    var canvas =3D $j('.large-thumb-canvas', el)=0A=
                    var prnt =3D  canvas.parent();=0A=
                    var win =3D $j(window)=0A=
=0A=
                    //=0A=
                    if (canvas && prnt && win)=0A=
                    {=0A=
=0A=
                        // collect necessary coordinates from all =
intresting objects=0A=
                        canvas.coords =3D this._x_coords(canvas, =
'canvas')=0A=
                        prnt.coords   =3D this._x_coords(prnt,   =
'parent')=0A=
                        win.coords    =3D this._x_coords(win,    =
'window')=0A=
=0A=
                        // replace @src with value of @hires, because =
href hold high resolution image and src low one.=0A=
                        $j('img', canvas).attr('src',=0A=
                                function(){=0A=
                                    var hires =3D $j(this).attr("hires");=0A=
                                    return (typeof (hires) !=3D =
'undefined' && hires !=3D this.src)=0A=
                                            ? hires : this.src=0A=
                                }=0A=
                        )=0A=
=0A=
                        // make arrangement of canvas in the view port =
nearby parent element.=0A=
                        var cnvsRltvToPrnt =3D =
this._x_arrange_canvas(canvas.coords, prnt.coords, win.coords)=0A=
=0A=
                        // visualize the canvas with images.=0A=
                        canvas.css('top', cnvsRltvToPrnt.top + 'px')=0A=
                        canvas.css('left', cnvsRltvToPrnt.left + 'px')=0A=
                        canvas.css('display', 'block')=0A=
                    } // canvas=0A=
                }, // over=0A=
                //=0A=
                out: function (el) {=0A=
                    var canvas =3D $j('.large-thumb-canvas', el)=0A=
                    canvas.css('display', 'none')=0A=
                },=0A=
                //=0A=
                _x_offset:                     function(el, elname) {=0A=
                    var c =3D {}=0A=
                    if (el[0] =3D=3D window )=0A=
                        c =3D this._x_scroll(el, elname)=0A=
                    else=0A=
                        c =3D el.offset()=0A=
                    //console.info('%s.offset {top,left} =3D {%s, %s}', =
elname, c.top, c.left)=0A=
                    return c=0A=
                },=0A=
                _x_scroll:                     function(el, elname) {=0A=
                    var c =3D {top: el.scrollTop(), left: =
el.scrollLeft()}=0A=
                    //console.info('%s.scroll {top,left} =3D {%s, %s}', =
elname, c.top, c.left)=0A=
                    return c=0A=
                },=0A=
                _x_width:                     function(el, elname) {=0A=
                    var w =3D el.outerWidth() || el.width()=0A=
                    //console.info('%s.width      =3D %s; outerWidth =3D =
%s; width =3D %s', elname, w, el.outerWidth(), el.width())=0A=
                    //IE: alert(elname + '.width      =3D ' + w + '; =
outerWidth =3D ' + el.outerWidth() + 'width =3D ' + el.width())=0A=
                    return w=0A=
                },=0A=
                _x_height:                     function(el, elname) {=0A=
                    var h =3D el.outerHeight() || el.height()=0A=
                    // console.info('%s.height =3D %s', elname, h)=0A=
                    return h=0A=
                },=0A=
                _x_coords:                    function(el, elname) {=0A=
                    var size =3D { width: this._x_width (el, elname), =
height:  this._x_height(el, elname)}=0A=
                    var coords =3D {=0A=
                            scroll:  this._x_scroll(el, elname),=0A=
                            width:   size.width,=0A=
                            height:  size.height,=0A=
                            offset:  this._x_offset(el, elname),=0A=
                            size:    size=0A=
                        }=0A=
                    return coords=0A=
                },=0A=
                _x_get_rltv_coord:            function(trgtObj, baseObj, =
label) {=0A=
                    var c =3D {top: trgtObj.top - baseObj.top, left: =
trgtObj.left - baseObj.left}=0A=
                    //console.info("%s {top: %s, left: %s}", label, =
c.top, c.left)=0A=
                    return c=0A=
                },=0A=
                //=0A=
                _x_get_empty_space:            function(objCoords, =
objSize, viewPortCoords, viewPortSize, label) {=0A=
                    var objRelToViewPort =3D =
this._x_get_rltv_coord(objCoords, viewPortCoords, '_x_get_empty_space: =
for ' + label);=0A=
                    var c =3D {above: objRelToViewPort.top,=0A=
                             below: viewPortSize.height - =
(objRelToViewPort.top + objSize.height),=0A=
                             left: objRelToViewPort.left,=0A=
                             right: viewPortSize.width - =
(objRelToViewPort.left + objSize.width)}=0A=
                    $j.each(c, function(k, v){c[k] =3D (v < 0 ? 0 : v)})=0A=
                    //console.info("%s {above: %s, below: %s, left: %s, =
right: %s}", label, c.above, c.below, c.left, c.right)=0A=
                    return c=0A=
                },=0A=
                //=0A=
                _x_get_bigest_space_area_name: function(spaces, label) {=0A=
                    var sArr =3D [];=0A=
                    for (k in spaces){ sArr[sArr.length] =3D {k: k, v: =
spaces[k]} }=0A=
                    // sort in reverse order=0A=
                    sArr.sort (function(a,b){return ( a.v < b.v ? 1 : ( =
a.v > b.v ?  - 1 : 0 ) )})=0A=
                    // console.info('%s: %s', label, sArr[0].k)=0A=
                    return sArr[0].k=0A=
                },=0A=
                //=0A=
                _x_get_space_ratios:             function(spaces, =
viewPortSize, label) {=0A=
                    var ratios =3D {above: 0, left: 0}=0A=
                    if (viewPortSize.height > 0)=0A=
                        ratios.above =3D spaces.above / =
viewPortSize.height=0A=
                    if (viewPortSize.width > 0)=0A=
                        ratios.left =3D spaces.left / viewPortSize.width=0A=
                    // console.info('ratio: %s {above: %s, left: %s}', =
label, ratios.above, ratios.left)=0A=
                    return ratios=0A=
                },=0A=
                _x_arrange_canvas:                function(c_c, prnt_c, =
viewp_c) {=0A=
                // then calculate empty space areas around parent =
element of the canvas, which is ususally an anchor ("a" element)=0A=
                    var emptySpacesArndPrntInViewPort =3D =
this._x_get_empty_space (prnt_c.offset, prnt_c.size, viewp_c.offset, =
viewp_c.size, 'emptySpacesArndPrntInWinPort')=0A=
=0A=
                    // get the area name, where the most linear space =
avaiable=0A=
                    var areaName =3D =
this._x_get_bigest_space_area_name(emptySpacesArndPrntInViewPort, =
viewp_c.size, 'spacios areaName')=0A=
=0A=
                    // calculate the proportions of area to be taken by =
canvas in empty space area=0A=
                    // relativly to the parent element.=0A=
                    var ratios =3D =
this._x_get_space_ratios(emptySpacesArndPrntInViewPort, viewp_c.size, =
'ratios')=0A=
=0A=
                    // calculate parent's coordinates relative to view =
port=0A=
                    var prntRltvToViewPort =3D =
this._x_get_rltv_coord(prnt_c.offset, viewp_c.offset, =
'prntRltvToViewPort')=0A=
=0A=
                    // declare canvas relative coordinates=0A=
                    var cnvsRltvToPrnt =3D {top: 0, left: 0}=0A=
=0A=
                    // choose appropriate location for placing canvas.=0A=
                    // and choose the 1st element of an array as the one =
with the biggest area around parent of canvas.=0A=
                    // console.info ('place it at the %s', =
bigestEmptySpaceAreaName)=0A=
                    // calculations are relative to the left corner of =
the parent element of canvas. (it has to have the css property =
position:relative)=0A=
                    switch (areaName)=0A=
                    {=0A=
                        case 'above':=0A=
                            cnvsRltvToPrnt.top  =3D - c_c.height=0A=
                            cnvsRltvToPrnt.left =3D prnt_c.width * 0.5 - =
c_c.width * ratios.left=0A=
                        break;=0A=
                        case 'below':=0A=
                            cnvsRltvToPrnt.top  =3D prnt_c.height=0A=
                            cnvsRltvToPrnt.left =3D prnt_c.width * 0.5 - =
c_c.width * ratios.left=0A=
                        break;=0A=
                        case 'left':=0A=
                            cnvsRltvToPrnt.left =3D - c_c.width=0A=
                            cnvsRltvToPrnt.top  =3D prnt_c.height * 0.5 =
- c_c.height * ratios.above=0A=
                        break;=0A=
                        case 'right':=0A=
                            cnvsRltvToPrnt.left =3D prnt_c.width=0A=
                            cnvsRltvToPrnt.top  =3D prnt_c.height * 0.5 =
- c_c.height * ratios.above=0A=
                        break;=0A=
                    }=0A=
=0A=
                    // recalculate canvas' coordinates relative to =
parent into coordinates relative to view port.=0A=
                    var cnvsRltvToViewPort =3D {=0A=
                        top:  prntRltvToViewPort.top + =
cnvsRltvToPrnt.top,=0A=
                        left: prntRltvToViewPort.left + =
cnvsRltvToPrnt.left=0A=
                    }=0A=
                    //console.info("cnvsRltvToPrnt {top:%s, left: %s}", =
cnvsRltvToPrnt.top, cnvsRltvToPrnt.left)=0A=
                    //console.info("cnvsRltvToViewPort {top:%s, left: =
%s}", cnvsRltvToViewPort.top, cnvsRltvToViewPort.left)=0A=
=0A=
                    // adjust relative position of canvas to the window =
if calculated values=0A=
                    // are forcing canvas go out of view port=0A=
                    // next order should be preserved to keep left top =
corner visible always=0A=
=0A=
                    // adjust right side and move a bit to the left=0A=
                    if (cnvsRltvToViewPort.left + c_c.width > =
viewp_c.width)=0A=
                    {=0A=
                         var horAdj =3D c_c.width - (viewp_c.width - =
cnvsRltvToViewPort.left)=0A=
                         cnvsRltvToPrnt.left     -=3D horAdj=0A=
                         cnvsRltvToViewPort.left -=3D horAdj=0A=
                         //console.log('adjust right side and move to =
the left %spx', horizontalAdjustment)=0A=
                    }=0A=
=0A=
                    // adjust below side and move to the top=0A=
                    if (cnvsRltvToViewPort.top + c_c.height > =
viewp_c.height)=0A=
                    {=0A=
                        var vertAdj =3D c_c.height - (viewp_c.height - =
cnvsRltvToViewPort.top)=0A=
                        cnvsRltvToPrnt.top     -=3D vertAdj=0A=
                        cnvsRltvToViewPort.top -=3D vertAdj=0A=
                        // console.log('adjust below side and move to =
the top %spx', verticalAdjustment)=0A=
                    }=0A=
=0A=
                    // adjust left side and move to the right=0A=
                    if (cnvsRltvToViewPort.left < 0)=0A=
                    {=0A=
                        // console.log('adjust left side and move to the =
right %spx', -(cnvsLeftRelativeToWindowPort))=0A=
                        cnvsRltvToPrnt.left    -=3D =
cnvsRltvToViewPort.left=0A=
                        cnvsRltvToViewPort.left =3D 0=0A=
                    }=0A=
                    // adjust above side and move to the bottom=0A=
                    if (cnvsRltvToViewPort.top < 0)=0A=
                    {=0A=
                        // console.log('adjust above side and move to =
the bottom %spx', -(cnvsTopRelativeToWindowPort))=0A=
                        cnvsRltvToPrnt.top          -=3D =
cnvsRltvToViewPort.top=0A=
                        cnvsRltvToViewPort.top =3D 0=0A=
                    }=0A=
                    // console.info("cnvsRltvToPrnt adjusted {top: %s, =
left: %s}", cnvsRltvToPrnt.top, cnvsRltvToPrnt.left)=0A=
                    // console.info("cnvsRltvToViewPort adjusted {top: =
%s, left: %s}", cnvsRltvToViewPort.top, cnvsRltvToViewPort.left)=0A=
                    return cnvsRltvToPrnt=0A=
                }=0A=
        })=0A=
         // =
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D  end of hoverHandlers =
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=0A=
=0A=
=0A=
         // =
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D  $j.fn.figPopup =
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D=3D=3D=3D=3D=0A=
        function  hoverOverHandler  () {=0A=
            var h =3D new hoverHandlers();=0A=
            h.over(this)=0A=
        }=0A=
=0A=
        function hoverOutHandler () {=0A=
            var h =3D new hoverHandlers();=0A=
            h.out(this)=0A=
        }=0A=
=0A=
        $j.fn.figPopup =3D function(opts){=0A=
            var config =3D {=0A=
                interval:      500,=0A=
                timeout:       0,=0A=
                over:          hoverOverHandler, // handler onMouseOver=0A=
                out:           hoverOutHandler    // handler  onMouseOut=0A=
            }=0A=
            $j.extend(config, opts)=0A=
            return  config=0A=
        }=0A=
=0A=
        // =
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
=3D  end of $j.fn.figPopup =
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=0A=
=0A=
    }) (jQuery) // end of extention of jQuery with figPopup=0A=
=0A=
=0A=
=0A=
    jQuery(document).ready( function () {=0A=
        var fp_cfg =3D new jQuery.fn.figPopup()=0A=
        jQuery('a.figpopup').hoverIntent(fp_cfg)=0A=
    });=0A=
}=0A=
=0A=
// end of hoverIntent routines for figures' popups=0A=
// =
*************************************************************************=
***=0A=
//  implementation of ....=0A=
// =
*************************************************************************=
***=0A=

------=_NextPart_000_0000_01CAD35F.003A3910
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.ncbi.nlm.nih.gov/corehtml/pmc/js/noext.menu.js

if (typeof(noext) =3D=3D=3D 'undefined') {
   noext =3D {};=20
}

if (typeof jQuery =3D=3D=3D 'undefined') {
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}

// Constructor
noext.Menu =3D function (n, subMenuStyle) {
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: 'opened';
    this.firstLi =3D this.n.children('li');
    this.subMenu =3D this.firstLi.find('ul');
    this.subMenuItems =3D this.subMenu.find('li');
    this.init();
    noext.Menu.menuInstances.push(this);
};
noext.Menu.menuInstances =3D [];

noext.Menu.prototype =3D {
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        var that =3D this;
        this.subMenuItems.mouseover( this.hover); // do mouseovers for =
ie6
        this.subMenuItems.mouseout( this.unHover);
        this.n.click( function (event) { that.update(event);}); // =
update the menu
       =20
        jQuery(document).click( function (event) { =
that.closeAllSubMenus(event); }); // close other menus when we click one
    },
    update: function (e) {
            var t =3D jQuery(e.target);         =20
            // don't follow link if first item, whose target is '<u>'
            if (t.hasClass('first-link') ) {
                e.preventDefault();
            }

            var sm =3D this.subMenu;
            // close other menus
            this.closeOtherSubMenus(sm);
      =20
            var subMenuStyle =3D this.subMenuStyle;
           =20
            if ( !sm.hasClass(subMenuStyle)) {
                sm.addClass(subMenuStyle);
            } else {
                sm.removeClass(subMenuStyle);
            }
            e.stopPropagation(); // don't let default event occur =
(removeMenus from document.click)

           =20

    },
    hover: function () {
        jQuery(this).addClass('active-item');

    },
    unHover: function () {
        jQuery(this).removeClass('active-item');

    },
    closeOtherSubMenus: function (currentSubMenu) {
        // close all menu's submenus, that are different than currently =
clicked menu's submenu
        var subMenuStyle =3D this.subMenuStyle;
        currentSubMenu =3D this.subMenu;
        var menuInstances =3D noext.Menu.menuInstances;
        for (var i =3D 0; i < menuInstances.length; i++) {
            var menuInstance =3D menuInstances[i];
    =20
            if (currentSubMenu !=3D=3D menuInstance.subMenu && =
menuInstance.subMenu.hasClass(subMenuStyle)) {
                menuInstance.subMenu.removeClass(subMenuStyle);
            }=20
        }
    },
    closeAllSubMenus: function (event) {
        // for when user clicks on document
        var menuInstances =3D noext.Menu.menuInstances;
        for (var i =3D 0; i < menuInstances.length; i++) {
            var subMenu =3D this.subMenu;
            subMenu.removeClass(this.subMenuStyle);
        }
    }
};
// end of menu objecta


jQuery(document).ready( function () {
    jQuery('ul.noext-menu').removeClass('noext-menu-hidden').each( =
function () {
        noext.Menu.menuInstances.push( new noext.Menu( jQuery(this) ) );
    });
});
------=_NextPart_000_0000_01CAD35F.003A3910
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.ncbi.nlm.nih.gov/portal/js/portal.js

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.doSubmitAttribute,null);}=0A=
oThis.listenForEvents(domCtrl,sActionEvent);}}finally{}};},addRule:functi=
on(oThis,sEvent,sName,fFunc,oArg){var rules=3Dthis._rules;var =
ename=3DsName+"$"+sEvent;var =
i;if(typeof(rules[ename])!=3D'undefined'){for(i=3D0;i<rules[ename].length=
;i++){if(rules[ename][i].func=3D=3DfFunc){return;}}}else{rules[ename]=3D[=
];}=0A=
rules[ename][rules[ename].length]=3D{'thisptr':oThis,'func':fFunc,'arg':o=
Arg};},getRulesFor:function(sName,sEvent){return =
this._rules[sName+"$"+sEvent];},listenForEvents:function(domCtrl,sEventNa=
me){domCtrl._i_=3D(domCtrl._i_||[]);if(!domCtrl._i_[sEventName]){domCtrl.=
_i_[sEventName]=3D1;utils.addEvent(domCtrl,sEventName,this.handleAction);=
}},doDataExchange:function(eEvent,domTarget,oArg){var =
d=3DDispatcher.getInstance();var rules=3DoArg;if(!rules){return;}=0A=
for(var =
k=3D0;k<rules.length;k++){d.notify(domTarget,rules[k].m,rules[k]);}},doSu=
bmitAttribute:function(eEvent,domTarget,oArg){eEvent.preventDefault();eEv=
ent.stopPropagation();console&&console.info("Submitting request because =
submit attribute set");var =
realname=3DdomTarget.getAttributeNode("realname");realname=3Drealname?rea=
lname.value:null;if(domTarget.name||realname){this.setSubmitSource(realna=
me||domTarget.name);}=0A=
if(domTarget.name=3D=3D"p$ExL"){el=3Ddocument.getElementsByName('p$el');i=
f((typeof(el)!=3D'undefined')&&(el.length>0)){el[0].value=3DdomTarget.get=
Attribute("href");}}=0A=
Dispatcher.getInstance().requestSubmit();},handleAction:function(e){var =
d=3DDispatcher.getInstance();var t=3Dthis;var =
i;if(t.tagName&&t.tagName.toLowerCase()=3D=3D'a'){e.preventDefault();e.st=
opPropagation();}=0A=
var =
realname=3Dthis.getAttributeNode("realname");realname=3Drealname?realname=
.value:null;if(this.name||realname){d.setSubmitSource(realname||this.name=
);}=0A=
d.submitCheckBegin();try{console&&console.info("Executing rule =
"+t.name+"."+e.type);var =
rules=3Dd.getRulesFor(t.name,e.type);for(i=3D0;rules&&(i<rules.length);i+=
+){var =
rule=3Drules[i];try{rule.func.call(rule.thisptr,e,t,rule.arg);}catch(err)=
{console&&console.error("Dispatcher.handleAction: rule failed: =
"+t.name+"$"+e.type+": =
"+err);}}}catch(err){console&&console.error("Error: handleAction: =
"+err);}finally{d.submitCheckEnd();}},actionEventName:function(t){var =
ttype=3D(t.getAttribute("type")||'').toLowerCase();switch(t.tagName.toLow=
erCase()){case'select':case'textarea':return'change';case'form':return'';=
case'input':if(ttype=3D=3D'text'){return'change';}}=0A=
return'click';},initHierarchy:function(oTree){if(!oTree||!oTree.name){ret=
urn;}=0A=
var =
sname=3DoTree.shortname||oTree.name.substring(1+oTree.name.lastIndexOf(".=
"));try{var =
c=3DPortal.Component.create(oTree.type,oTree.name,sname,this.oNotifier);c=
.realname=3DoTree.realname;}catch(e){console&&console.group&&console.grou=
p("Exception creating portlet =
"+oTree.name);console&&console.info("Message: =
"+e.toString());if(typeof(e)!=3D'string'){console&&console.group&&console=
.group("Details");console&&console.dir&&console.dir(e);console&&console.g=
roupEnd&&console.groupEnd();}=0A=
console&&console.group&&console.group("Trace")=0A=
console&&console.trace&&console.trace();console&&console.groupEnd&&consol=
e.groupEnd();console&&console.groupEnd&&console.groupEnd();}=0A=
if(oTree.children){for(var =
i=3D0;i<oTree.children.length;i++){this.initHierarchy(oTree.children[i]);=
}}},submit:function(){if(this.submitCheck()){if(xmlHttpCall&&xmlHttpCall.=
abortCalls){xmlHttpCall.abortCalls();}=0A=
document.forms[0]._submit();}},submitCheck:function(){var =
oThis=3Dthis;var =
thisForm=3Ddocument.forms[0];this.submitOK=3Dtrue;this.vetoWhy=3Dnull;thi=
s.notify(this,"portal$beforesubmit",{form:thisForm},null);return =
this.submitOK;},veto:function(whydata){this.submitOK=3Dfalse;this.vetoWhy=
=3Dwhydata;},isVetoed:function(){return!this.submitOK;},setSubmitSource:f=
unction(sourceName){var =
el=3Ddocument.getElementsByName('p$a');if((typeof(el)!=3D'undefined')&&(e=
l.length>0)){if(typeof(sourceName)=3D=3D'undefined'){console&&console.war=
n("Warning: Can't identify submitter: using =
p$a=3D''");}else{el[0].value=3DsourceName;}}},getSrcDst:function(oRule,oN=
otifierObj){function x_FindObj(name,sid){var oResult=3D[];var =
oControls=3D$N(name);for(var =
i=3D0;i<oControls.length;i++){if("*"=3D=3Dsid||oControls[i].getAttribute(=
"sid")=3D=3Dsid){oResult[oResult.length]=3DoControls[i];}}=0A=
return oResult;}=0A=
var oThis=3Dthis;var name=3DoRule.sname;var =
sid=3DoRule.ssid;if("#"=3D=3Dname){name=3DoRule.ename;}=0A=
if("#"=3D=3Dsid){sid=3DoRule.esid;sid=3DoNotifierObj.getAttribute("sid");=
}=0A=
var =
oSrc=3Dx_FindObj(name,sid);name=3DoRule.dname;sid=3DoRule.dsid;if("#"=3D=3D=
name){name=3DoRule.ename;}=0A=
if("#"=3D=3Dsid){sid=3DoRule.esid;sid=3DoNotifierObj.getAttribute("sid");=
}=0A=
var =
oDst=3Dx_FindObj(name,sid);return{src:oSrc,dst:oDst};},setListener:functi=
on(oListener,sMessage,fFunction,oNotifier){return =
this.oNotifier.setListener(oListener,sMessage,fFunction,oNotifier);},noti=
fy:function(oNotifier,sMessage,oComment,oListener){return =
this.oNotifier.Notify(oNotifier,sMessage,oComment,oListener);},dataExchan=
geFunctions:{CopyValue:function(oListener,oRule,sMessage,oNotifierObj){va=
r dispatcher=3DoListener;var =
oSrcDst=3Ddispatcher.getSrcDst(oRule,oNotifierObj);for(var =
j=3D0;j<oSrcDst.dst.length;j++){for(var =
i=3D0;i<oSrcDst.src.length;i++){oSrcDst.dst[j].value=3DoSrcDst.src[i].val=
ue;}}},AddValue:function(oListener,oRule,sMessage,oNotifierObj){var =
dispatcher=3DoListener;var =
oSrcDst=3Ddispatcher.getSrcDst(oRule,oNotifierObj);for(var =
j=3D0;j<oSrcDst.dst.length;j++){for(var =
i=3D0;i<oSrcDst.src.length;i++){oSrcDst.dst[j].value+=3D(oSrcDst.dst[j].v=
alue>""?", =
":"")+oSrcDst.src[i].value;}}},SetValue:function(oListener,oRule,sMessage=
,oNotifierObj){var dispatcher=3DoListener;var =
oSrcDst=3Ddispatcher.getSrcDst(oRule,oNotifierObj);for(var =
j=3D0;j<oSrcDst.dst.length;j++){var =
dst=3DoSrcDst.dst[j];if(Dispatcher.isCheckable(dst)){dst.checked=3DoRule.=
p[0];}else{dst.value=3DoRule.p[0];}}},ItemToList:function(oListener,oRule=
,sMessage,oNotifierObj){var dispatcher=3DoListener;var =
oSrcDst=3Ddispatcher.getSrcDst(oRule,oNotifierObj);for(var =
j=3D0;j<oSrcDst.dst.length;j++){var dst=3DoSrcDst.dst[j];var =
dstval=3Ddst.value.replace(/^\s+/,"").replace(/\s+$/,"");var =
dstItems=3Ddstval.split(/\s*,\s*/).filter(function(s){return =
s!=3D=3D"";});for(var i=3D0;i<oSrcDst.src.length;i++){var =
src=3DoSrcDst.src[i];var sc=3DDispatcher.isCheckable(src);var =
checked=3D(sc?src.checked:(src.value!=3D=3D""));var =
position=3DdstItems.indexOf(src.value);if(checked&&position<0){dstItems[d=
stItems.length]=3Dsrc.value;}=0A=
if(!checked&&position>=3D0){dstItems.splice(position,1);}}=0A=
dst.value=3DdstItems.join(", =
");}},PropertyToValue:function(oListener,oRule,sMessage,oNotifierObj){var=
 dispatcher=3DoListener;var =
oSrcDst=3Ddispatcher.getSrcDst(oRule,oNotifierObj);for(var =
j=3D0;j<oSrcDst.dst.length;j++){for(var =
i=3D0;i<oSrcDst.src.length;i++){oSrcDst.dst[j].value=3DoSrcDst.src[i].get=
Attribute(oRule.p[0]);}}},PropertyToList:function(oListener,oRule,sMessag=
e,oNotifierObj){var dispatcher=3DoListener;var =
oSrcDst=3Ddispatcher.getSrcDst(oRule,oNotifierObj);for(var =
j=3D0;j<oSrcDst.dst.length;j++){var dst=3DoSrcDst.dst[j];var o=3D{};var =
values=3D[];var propname=3DoRule.p[0];for(var =
i=3D0;i<oSrcDst.src.length;i++){var src=3DoSrcDst.src[i];var =
prop=3Dsrc.getAttribute(propname);if(!prop){continue;}=0A=
var =
sc=3DDispatcher.isCheckable(src);if(sc?src.checked:(prop&&prop!=3D=3D""))=
{if(!o[prop]){values[values.length]=3Dprop;}=0A=
o[prop]=3D1;}}=0A=
dst.value=3Dvalues.join(", =
");}}},requestSubmit:function(src){console&&console.info("Request =
submitted");this.submitter=3Dsrc;this.submitRequested=3Dtrue;},submitChec=
kBegin:function(){if(this.submitChecking){throw"ERRINTERNAL: recursive =
submit check";}=0A=
this.submitChecking=3Dtrue;this.submitter=3Dnull;this.submitRequested=3Df=
alse;},submitCheckEnd:function(){if(this.submitRequested){this.submit();}=0A=
this.submitChecking=3Dfalse;this.submitter=3Dnull;this.submitRequested=3D=
false;}},{_instance:null,getInstance:function(){return =
Dispatcher._instance||(Dispatcher._instance=3Dnew =
Dispatcher());},onload:function(){var sc=3D"Create components";var =
si=3D"Initialize components";var =
d=3DDispatcher.getInstance();d.initHierarchy(objHierarchy);d.initDataExch=
ange(ObjectLinks);d.setListener(d,'portal$vetosubmit',function(dst,data,m=
essage,src){d.veto(data);},null);var =
theForm=3Ddocument.forms[0];if(theForm&&!theForm._submit){theForm._submit=
=3DtheForm.submit;theForm.submit=3Dfunction(){Dispatcher.getInstance().re=
questSubmit();};}},isCheckbox:function(node){var =
t=3Dnode.getAttribute("type");return =
t&&(t.toLowerCase()=3D=3D"checkbox");},isRadio:function(node){var =
t=3Dnode.getAttribute("type");return =
t&&(t.toLowerCase()=3D=3D"radio");},isCheckable:function(node){return =
this.isRadio(node)||this.isCheckbox(node);}});utils.addEvent(window,"load=
",Dispatcher.onload,false);Portal=3DBase.extend({constructor:function(){t=
his.init();},init:function(){}},{_instance:null,getInstance:function(){re=
turn Portal._instance||(Portal._instance=3Dnew =
Portal());},requestSubmit:function(){Dispatcher.getInstance().requestSubm=
it();},$send:function(sMessage,oData,oDst){var =
d=3DDispatcher.getInstance();d.submitCheckBegin();d.notify(null,sMessage,=
oData,oDst);d.submitCheckEnd();}});Portal.Component=3DBase.extend({constr=
uctor:function(path,name,notifier){this.init(path,name,notifier);},init:f=
unction(path,name,notifier){this.name=3Dname;this.path=3Dpath;this.notifi=
er=3Dnotifier;if(this.listen){var msg;var isEvent=3Dnew =
RegExp(/^([^<]+)<([^>]+)>/);var m;var =
d=3DDispatcher.getInstance();for(msg in =
this.listen){m=3DisEvent.exec(msg);if(m){this.addEvent(m[1],m[2],this.lis=
ten[msg],false);}else{this._listen(msg,this.listen[msg],null);}}}=0A=
if(this.send){for(msg in =
this.send){if(this.send[msg]=3D=3Dnull){this.send[msg]=3Dthis.makeSender(=
this,msg);}}}},makeSender:function(sender,msg){return =
function(obj){return =
sender._send(msg,obj,null);};},getValue:function(attr){var =
prop=3Dnull;var ix;if(typeof =
attr=3D=3D=3D"object"){prop=3Dattr.prop;attr=3Dattr.attr;}=0A=
else =
if((ix=3Dattr.indexOf(":"))>=3D0){prop=3Dattr.substring(ix+1);attr=3Dattr=
.substring(0,ix);}=0A=
var inp=3Dthis.getInputs(attr);if(!inp){return null;}=0A=
if(inp.length=3D=3D1){inp=3Dinp[0];return(prop&&prop.toLowerCase()!=3D=3D=
"value")?inp.getAttribute(prop):htmlutils.getValue(inp);}=0A=
var result=3D[];for(var =
i=3D0;i<inp.length;i++){if(prop&&prop.toLowerCase()!=3D=3D"value"){if(typ=
eof(inp[i][prop])!=3D'undefined'){result[result.length]=3Dinp[i][prop];}e=
lse{result[result.length]=3Dinp[i].getAttribute(prop);}}else{result[resul=
t.length]=3Dhtmlutils.getValue(inp[i]);}}=0A=
return result;},getList:function(attr){var ix;var =
prop=3Dnull;if((ix=3Dattr.indexOf(":"))>=3D0){prop=3Dattr.substring(ix+1)=
;attr=3Dattr.substring(0,ix);}=0A=
if(prop){throw"UNIMPLEMENTED: Component.getList: Getting list by =
property";}=0A=
var inp=3Dthis.getInputs(attr);if(!inp){return null;}=0A=
if(inp.length=3D=3D1){inp=3Dinp[0];return htmlutils.getList(inp);}=0A=
var result=3D[];for(var =
i=3D0;i<inp.length;i++){result[result.length]=3Dhtmlutils.getList(inp[i])=
;}=0A=
return result;},setValue:function(attr,value){var prop=3Dnull;var =
ix;if((ix=3Dattr.indexOf(":"))>=3D0){prop=3Dattr.substring(ix+1);attr=3Da=
ttr.substring(0,ix);}=0A=
var =
inp=3Dthis.getInputs(attr);if(inp.length=3D=3D1){inp=3Dinp[0];if(prop){if=
(typeof(inp[prop])!=3D'undefined'){inp[prop]=3Dvalue;}else{inp.setAttribu=
te(prop,value);}}else{inp.value=3Dvalue;}}else{throw"UNIMPLEMENTED: =
Cannot (yet) set vector values from =
scalar";}},getInputs:function(name){var =
inp=3Ddocument.getElementsByName(this.path+"."+name);if(!inp||inp.length=3D=
=3D=3D0){inp=3Dnull;}=0A=
return inp;},getInput:function(name){var =
inp=3Dthis.getInputs(name);return((inp&&inp.length>0)?inp[0]:null);},has:=
function(attrname){var =
inp=3Ddocument.getElementsByName(this.path+"."+attrname);return(inp&&(inp=
.length>0));},addEvent:function(inputName,eventName,f,flag){var =
inputs;var =
oThis=3Dthis;if(typeof(inputName)=3D=3D'string'){inputs=3Dthis.getInputs(=
inputName);}else =
if(utils.isArray(inputName)){inputs=3DinputName;}else{inputs=3D[inputName=
];}=0A=
if(!inputs){console&&console.warn("Can't find: "+inputName);return;}=0A=
var =
d=3DDispatcher.getInstance();for(i=3D0;i<inputs.length;i++){d.addRule(oTh=
is,eventName,inputs[i].name,f,null);d.listenForEvents(inputs[i],eventName=
);}=0A=
return;},removeEvent:function(inputName,eventName,f,flag){var =
inputs=3Dthis.getInputs(inputName);for(var =
i=3D0;inputs&&(i<inputs.length);i++){utils.removeEvent(inputs[i],eventNam=
e,f,flag);}},iListen:0,iNextListen:function(){return"L$"+this.iListen++;}=
,_listen:function(sMessage,fFunction,sSource){var oThis=3Dthis;var =
fName=3Dthis.iNextListen()+"$"+sMessage;this[fName]=3DfFunction;var =
d=3DDispatcher.getInstance();d.setListener(oThis,sMessage,function(dst,da=
ta,message,src){oThis[fName](message,data,src);},sSource);},_send:functio=
n(sMessage,oData,oDst){var =
d=3DDispatcher.getInstance();d.notify(this,sMessage,oData,oDst);},getMeta=
Data:function(){var =
result=3D{};result.listensFor=3D[];if(this.listen){for(var msg in =
this.listen){result.listensFor[result.listensFor.length]=3Dmsg;}}=0A=
result.sends=3D[];if(this.send){for(msg in =
this.send){result.sends[result.sends.length]=3Dmsg;}}=0A=
result.attributes=3D[];for(var attr in =
this.getAttributes()){result.attributes[result.attributes.length]=3Dattr;=
}=0A=
return =
result;}},{$i:{},$$i:{},create:function(type,path,cname,notifier){var =
p=3DPortal.Component.$i;var pelem=3Dpath.split(".");for(var =
i=3D0;i<pelem.length-1;i++){p=3Dp[pelem[i]];}=0A=
var =
sname=3Dpath.substring(1+path.lastIndexOf("."));if(typeof(Portal[type])=3D=
=3D'undefined'){throw("No such portlet type: "+type);}=0A=
if(typeof(Portal[type][sname])=3D=3D'undefined'){p[sname]=3Dnew =
Portal[type](path,cname,notifier);}else{try{p[sname]=3Dnew =
Portal[type][sname](path,cname,notifier);}catch(e){console&&console.error=
("Creating Portlet."+type+"."+sname+": "+e.message);throw e;}}=0A=
var =
l;if(!(l=3DPortal.Component.$$i[sname])){l=3DPortal.Component.$$i[sname]=3D=
[];}=0A=
l[l.length]=3Dp[cname];if(p[cname].beforesubmit){var =
d=3DDispatcher.getInstance();var =
oThis=3Dthis;d.setListener(oThis,"portal$beforesubmit",function(dst,data,=
message,src){var =
whydata=3Dp[cname].beforesubmit();if(typeof(whydata)=3D=3D'boolean'){retu=
rn whydata;}else =
if(typeof(whydata)=3D=3D'object'){d.notify(this,'portal$vetosubmit',whyda=
ta,null);return true;}=0A=
throw cname+".beforesubmit: returned "+typeof(whydata);},null);}=0A=
return p[cname];},get:function(path){return =
eval("Portal.Component.$i."+path);},getByShortName:function(name){return =
eval("Portal.Component.$$i."+name);},classExists:function(name){return(ty=
peof(Portal.Component[name])!=3D"undefined");},instanceExists:function(pa=
th){try{eval("Portal.Component.$i"+path);return true;}catch(e){}=0A=
return =
false;}});Portal.Layout=3DPortal.Component.extend({constructor:function(p=
ath,name,notifier){this.base(path,name,notifier);}});Portal.Cluster=3DPor=
tal.Component.extend({constructor:function(path,name,notifier){this.base(=
path,name,notifier);}});Portal.Portlet=3DPortal.Component.extend({getAttr=
ibutes:function(){var t=3Ddocument.getElementsByTagName("input");var =
result=3D{};var pathlen=3Dthis.path.length;for(var =
i=3D0;i<t.length;i++){var it=3Dt[i];var =
itn=3Dit.getAttribute("name");if(!itn){continue;}=0A=
var prefixlen=3Ditn.lastIndexOf(".");var =
samePrefix=3D(itn.substring(0,this.path.length)=3D=3Dthis.path);if((prefi=
xlen=3D=3Dpathlen)&&samePrefix){result[itn.substring(1+prefixlen)]=3Dit.v=
alue;}}=0A=
return result;}});$PI=3Dfunction(path){return =
Portal.Component.get(path);};$PN=3Dfunction(name){var =
c=3DPortal.Component.getByShortName(name);if(typeof(c)=3D=3D"undefined"){=
return null;}=0A=
return(c.length=3D=3D1)?c[0]:c;};htmlutils=3D{accessors:{select:{getValue=
:function(node){return this.getList(node).join(", =
");},getList:function(node){var opts=3Dnode.options;var r=3D[];for(var =
i=3D0;i<opts.length;i++){if(opts[i].selected){r[r.length]=3Dopts[i].value=
;}}=0A=
return r;}},button:{getValue:function(node){var =
attrNode=3Dnode.getAttributeNode('value');return(attrNode!=3D=3Dnull)?att=
rNode.nodeValue:null;}}},getValue:function(elem){var =
v=3Dthis.accessors[elem.tagName.toLowerCase()];return =
v&&v.getValue?v.getValue(elem):elem.value;},getList:function(elem){var =
v=3Dthis.accessors[elem.tagName.toLowerCase()];return =
v&&v.getList?v.getList(elem):(elem.value?[elem.value]:[]);}};utils.addEve=
nt(window,"load",function(){Portal.getInstance();},false);
------=_NextPart_000_0000_01CAD35F.003A3910
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.ncbi.nlm.nih.gov/portal/portalrc.fcgi/40595/js/15394/28968/29966/27166/30783/28669/27885/27590/29994/24519/29669/31945.js

//=0A=
//		 PAFAppResources: Initialize client side of Portlet Application =
Framework=0A=
//=0A=
=0A=
Portal.Portlet.PAFAppResources =3D Portal.Portlet.extend({=0A=
=0A=
   // In PAF, the form always posts to itself. It should update and =
interpret data, and=0A=
   // then *redirect* to another page if necessary.=0A=
   =0A=
   // This code fixes up the EntrezForm in PAF templates to =
automatically post=0A=
   // to the current window pathname.=0A=
   =0A=
   // If the programmer specifies a hidden input with =
id=3D"entrez-action",=0A=
   // that action (instead of POST) is used for the form.=0A=
   =0A=
   // If the programmer specifies a hidden input with =
id=3D"entrez-method",=0A=
   // that method is used for the form. =0A=
       =0A=
   // FIXME: This is a hack. EntrezForm should default to posting to =
same page,=0A=
   // and there should be some way of indicating how to post (or get) =
elsewhere.=0A=
   // The problem here is that we've mixed Entrez into PAF. This should =
be refactored.=0A=
   =0A=
   init: function(path, name, notifier) {=0A=
      var oThis =3D this;  // Use <oThis> instead of <this> for =
registering callbacks=0A=
      this.base(path, name, notifier);  // Call superclass constructor=0A=
      =0A=
      if (!document.forms[0]) return; // Do nothing if no form.=0A=
      =0A=
      var getNodeValue =3D function(id, dfl) {=0A=
         var node =3D $(id);=0A=
         return node ? node.value : dfl;=0A=
      };=0A=
      =0A=
      var entrezAction =3D getNodeValue('entrez-action', =
window.location.pathname);=0A=
      var entrezMethod =3D getNodeValue('entrez-method', null);=0A=
      var entrezSubmit =3D getNodeValue('entrez-submit', false);=0A=
      =0A=
      document.forms[0].action =3D entrezAction;=0A=
      =0A=
      // FIXME: We need to include query string here, too, I think... At =
least for POST...      =0A=
      if (entrezMethod) {=0A=
         document.forms[0].method =3D entrezMethod;=0A=
      }=0A=
=0A=
      // By default, entrez form ignores submits. Setting "entrezSubmit" =
to "true" means let Entrez form=0A=
      // actually process the submit. This probably will only work =
correctly for action=3D"get"       =0A=
      if (entrezSubmit) {=0A=
         document.forms[0].onsubmit=3D"return true;"=0A=
      }=0A=
   }=0A=
});=0A=
=0A=
// makes an ext js looking portlet w/o using extjs
if (typeof(noext) =3D=3D 'undefined') {
    noext =3D {};
}
noext.parseConfig =3D function(n) {
    var configStr =3D n.getAttribute('config') || '';
    try {
        var jsonObj =3D eval('({' + configStr + '})');
    }
    catch(e) {
        if (window.console) {
            console.error(e.message + ' in local config object in =
noext.Portlet: file: ' + e.fileName + ', line ' + e.lineNumber + '.' );
        }
    }
    return jsonObj;
};
noext.getClassArray =3D function(n) {
    var classNameStr =3D n.className;
    return classNameStr.split(' ');
};
noext.getFirstHeader =3D function(n) {
    var fc =3D utils.getFirstChild(n);
    if(fc.tagName.search(/^h\d/i) !=3D -1) {
        return fc;
    }
};
noext.Portlet =3D function(n, beenClassed) {
    this.n =3D n;=0A=
    this.beenClassed =3D beenClassed;=0A=
    if (!beenClassed) {
        this.headerNode =3D noext.getFirstHeader(this.n);
        this.headerDiv =3D document.createElement('div');
        this.bodyDiv =3D document.createElement('div');
        utils.addClass(this.bodyDiv, 'x-panel-body');=0A=
    }=0A=
    else {=0A=
        var divNodes =3D n.getElementsByTagName("div");=0A=
        for (var i =3D 0; i < divNodes.length; ++i) {=0A=
            var divNode=3D divNodes[i];=0A=
            if (utils.hasClass(divNode, 'x-panel-header')) {=0A=
                this.headerDiv =3D divNode;=0A=
            }=0A=
            else if (utils.hasClass(divNode, 'x-panel-body')) {=0A=
                this.bodyDiv =3D divNode;=0A=
            }=0A=
        }=0A=
    }
    var configObj =3D noext.parseConfig(this.n);
    this.collapsible =3D configObj.collapsible =3D=3D false ? false : =
true;=20
    this.closeable =3D configObj.closeable =3D=3D false ? false : true;=20
    this.collapsed =3D configObj.collapsed =3D=3D true ? true : false;
    if (this.collapsed =3D=3D true) {
        utils.addClass(this.n, 'x-panel-collapsed');
        this.bodyDiv.style.display =3D 'none';
    }
    this.make();
};

noext.Portlet.instances =3D [];
noext.Portlet.ready =3D function() {
    var nodes =3D $C('port', 'class', document, 'div');
    for (var i =3D 0; i < nodes.length; i++) {
        var n =3D nodes[i];
        if (utils.hasClass(n, 'norender')) {
            continue;
        }=0A=
        // CFM:  If the top-level div already has the 'x-panel' class, =
then =0A=
        // assume that it has been completely "classed up".=0A=
        var beenClassed =3D utils.hasClass(n, 'x-panel');
        portlet =3D new noext.Portlet(n, beenClassed);
    }
}
noext.Portlet.prototype =3D {
    make: function() {=0A=
        var beenClassed =3D this.beenClassed;=0A=
        =0A=
        var toolCloseA, toggleA;=0A=
        if (!beenClassed) {
            // put a parent around body content
            //var innerContainingDiv =3D document.createElement('div');
            utils.addClass(this.n, 'x-panel');
            //this.n.style.width =3D 'auto';
            utils.addClass(this.headerDiv, 'x-panel-header');
            utils.addClass(this.headerDiv, 'x-unselectable');
            if (this.headerNode) {=20
                this.n.insertBefore(this.headerDiv, this.headerNode);
                var header =3D this.n.removeChild(this.headerNode);
            }
            toolCloseA =3D document.createElement('a');
            toolCloseA.setAttribute('href', '#');
            utils.addClass(toolCloseA, 'x-tool');
            utils.addClass(toolCloseA, 'x-tool-close');
            toggleA =3D document.createElement('a');
            toggleA.setAttribute('href', '#');
            utils.addClass(toggleA, 'x-tool');
            utils.addClass(toggleA, 'x-tool-toggle');
            if (this.closeable =3D=3D true) {
                this.headerDiv.appendChild(toolCloseA);
            }
            if (this.collapsible) {
                this.headerDiv.appendChild(toggleA);
            }
            if (this.headerNode) {
                this.headerDiv.appendChild(header);
                utils.addClass(this.headerNode, 'x-panel-header-text');
            }=0A=
        }=0A=
        else {=0A=
            // Find the close and toggle <a> elements, if they exist.  =
To do this, we=0A=
            // iterate through all the <a> children, looking for the =
right classes.=0A=
            var aKids =3D this.n.getElementsByTagName("a");=0A=
            for (var i =3D 0; i < aKids.length; ++i) {=0A=
                var aKid =3D aKids[i];=0A=
                if (utils.hasClass(aKid, 'x-tool-close')) {=0A=
                    toolCloseA =3D aKid;=0A=
                }=0A=
                else if (utils.hasClass(aKid, 'x-tool-toggle')) {=0A=
                    toggleA =3D aKid;=0A=
                }=0A=
            }=0A=
        }=0A=
        this.toolCloseA =3D toolCloseA;
        this.toggleA =3D toggleA;=0A=
        if (!beenClassed) {
            this.reParent();=0A=
        }
        this.setEvents();
    },=20
    reParent: function() {
        var bodyElems =3D [];
        utils.insertAfter(this.n, this.bodyDiv, this.headerDiv);
        for (var i =3D this.n.childNodes.length -1; i >=3D 0; i--) {
            var fc =3D this.n.childNodes[i];
            if (fc !=3D this.headerDiv && fc !=3D this.bodyDiv) {=20
                bodyElems.push(this.n.removeChild(fc));
            }
            for (var j =3D 0; j < bodyElems.length; j++) {
                this.bodyDiv.appendChild(bodyElems[j]);
            }
        }
    },
    setEvents: function(toggleA) {
        var that =3D this;=0A=
        if (this.toggleA) {
            utils.addEvent(this.toggleA, 'click', function(e) { =
that.toggle(e)});=0A=
        }=0A=
        if (this.toolCloseA) {
            utils.addEvent(this.toolCloseA, 'click', function(e) { =
that.close(e)});=0A=
        }
    },=20
    toggle: function(e) {
        utils.preventDefault(e);
        if (utils.hasClass(this.n, 'x-panel-collapsed')) {
            utils.removeClass(this.n, 'x-panel-collapsed')
            this.bodyDiv.style.display =3D 'block';
        } else {
            utils.addClass(this.n, 'x-panel-collapsed');
            this.bodyDiv.style.display =3D 'none';
        }
    },
    close: function(e) {
        utils.preventDefault(e);=20
        this.n.parentNode.removeChild(this.n);
    }
};

utils.addEvent(window, 'load', noext.Portlet.ready);
=0A=
// This 2.x branch of this file was to add a brief delay to the pop-up, =
but it=0A=
// was decided not to merge this change into the main trunk.  I.e. this =
branch=0A=
// is PMC specific.  See PP-207.=0A=
=0A=
Portal.Portlet.BriefLinkPageSection =3D Portal.Portlet.extend({

	init: function(path, name, notifier) {=0A=
	    console.info("Created BriefLinkPageSection");
		this.base(path, name, notifier);
        =0A=
        this.SetPortletName();=0A=
        if (Portal.Portlet.BriefLinkPageSection.portletname !=3D ''){=0A=
            Portal.Portlet.BriefLinkPageSection.portletname =3D "." + =
Portal.Portlet.BriefLinkPageSection.portletname;=0A=
        }=0A=
        =0A=
        this.CreatePopNodes();=0A=
       =20
	},=0A=
	=0A=
	SetPortletName: function(){=0A=
	    // derived portlet should override and provide portlet name;=0A=
	},=0A=
	=0A=
    popNodeDelay: 0,
=0A=
    CreatePopNodes: function(){=0A=
        var pop =3D cssQuery("div.brieflink div" + =
Portal.Portlet.BriefLinkPageSection.portletname + " ul");
        var poptype =3D pop[0].className;
       =20
        // when value is pop0
        if (poptype.match(/pop0/)){
            // no popnodes need to be created
        }
        // for all other cases try to create popnode
        else{
            // Create ElementPopper for each li.item
    		var elemid =3D "div.brieflink div" + =
Portal.Portlet.BriefLinkPageSection.portletname + " li.item";
    		var elements =3D cssQuery(elemid);=20
    	=09
    		// go through the list of elements
    		for (var i =3D 0; i < elements.length; i++) {
                var elem =3D elements[i];
                var content =3D "";=20
               =20
                // when value is pop1
                if (poptype.match(/pop1/)){
                    // create pop with title + desc
                    var linktext =3D cssQuery("a", elem);
                    if (linktext && linktext.length > 0){
                        content +=3D linktext[0].innerHTML + "<br/>";
                    }
                }
                // default case - when value is pop2 or anything else =
but pop3
                else if (!poptype || !poptype.match(/pop3/)){
                    // if visible content length is larger than 105, =
then show the entire text in popup,
                    // followed by any included link description.
                    var linktext =3D cssQuery("a", elem);
                    var note =3D cssQuery("p.note", elem);
                    var content =3D "";=20
                    var linklength =3D 0;
                    if (linktext && linktext.length > 0){
                       linklength +=3D linktext[0].innerHTML.length;
                    }
                    if (note && note.length > 0){
                       linklength +=3D note[0].innerHTML.length;
                    }
                    if (linklength > 100){
                       content +=3D linktext[0].innerHTML + "<br/>";
                    }
                } // end default case
	          =20
	          =20
                // add Description to pop node when it exists for all =
popnode cases	          =20
	            var desc =3D cssQuery("p.desc", elem);
                if (desc && desc.length > 0){
                   content +=3D desc[0].innerHTML;
                }
               =20
                // create popnode when content is present
                if (content && content.length > 0) {
                  var elemclass =3D elem.className;
                  if (!elemclass.match(/popnode/)){
                      elem.className =3D elemclass + " popnode";
                  }
                  //new ElementPopper(elem, content);
                  new ElementPopper(elem, {=20
                      content: content,
                      delay: this.popNodeDelay=20
                  });
                }
               =20
            } // end for=20
        } // end else=0A=
	} // end CreatePopNodes function
},=0A=
{=0A=
portletname: '' =0A=
=0A=
});=0A=
//=0A=
// Java Module PopDiv_JS=0A=
//=0A=
=0A=
//=0A=
// Shows and hides a div at a specific location, either (x,y), or =
relative to=0A=
// an element in a specific direction.=0A=
//=0A=
// Limitations:=0A=
//  - height or width must be set on the popper div, or it may not =
position correctly.=0A=
//=0A=
// You're probably better off using ElementPopper instead of PopDiv for =
hover pops. Popper handles mouse events=0A=
// correctly.=0A=
//=0A=
// Arguments:=0A=
//   divId: the id of the div to use for the popper. You can also pass a =
DOM node, and PopDiv will use that node.=0A=
//       The default is "popperDiv".=0A=
//   className: Add this className to the popper div. Optional--no =
default. This is useful for setting up default state.=0A=
//=0A=
// It's preferable to use a single PopDiv to render popups all over=0A=
// a single page. You only need as many PopDivs as the maximum number of =
popups you may=0A=
// ever have open at once. See ElementPopper for details.=0A=
=0A=
var PopDiv =3D function(divId, className) {=0A=
=0A=
   var innerdiv =3D null;=0A=
   =0A=
   // default is a string=0A=
   if (typeof(divId) =3D=3D 'undefined') {=0A=
      divId =3D "popperDiv";=0A=
   }=0A=
   =0A=
   // Find or make popper div=0A=
   if ((this.div =3D document.getElementById(divId)) =3D=3D null) {=0A=
   =0A=
      this.div =3D document.createElement("div");=0A=
      this.div.id =3D divId;=0A=
      this.div.className =3D "popper";=0A=
      if (className) {=0A=
         this.div.className +=3D " " + className;=0A=
      }=0A=
      document.body.appendChild(this.div);=0A=
      this.div.innerHTML =3D "<div class=3D'popperInnerDiv'/>";=0A=
   } else {=0A=
      utils.addClass(this.div, "popper"); // Be sure it has this=0A=
   }=0A=
=0A=
   // this.innerdiv is first element of this.div=0A=
   for (this.innerdiv =3D this.div.firstChild;=0A=
      this.innerdiv && this.innerdiv.nodeType !=3D 1;=0A=
      this.innerdiv =3D this.innerdiv.nextSibling) {=0A=
   }=0A=
   =0A=
   // If caller provided a div with no inner divs, add one.=0A=
   if (!this.innerdiv) {=0A=
       this.innerdiv =3D document.createElement("div");=0A=
       this.innerdiv.className =3D "popperInnerDiv";=0A=
       this.div.appendChild(this.innerdiv);=0A=
   }=0A=
}=0A=
=0A=
PopDiv.POP_ABOVE =3D 0;=0A=
PopDiv.POP_RIGHT =3D 1;=0A=
PopDiv.POP_BELOW =3D 2;=0A=
PopDiv.POP_LEFT =3D 3;=0A=
PopDiv.POP_CLASSES =3D ['pop-above', 'pop-right', 'pop-below', =
'pop-left'];=0A=
=0A=
PopDiv.POPPED =3D 'popped';=0A=
PopDiv.POP_GETXY =3D 'pop-getxy';=0A=
=0A=
PopDiv.prototype =3D {=0A=
   // Show it at x, y=0A=
   showAt: function(html, x, y) {=0A=
=0A=
    //console.info("POP @ (" + x + ", " + y + ")");=0A=
    //console.info("----");=0A=
    =0A=
      // Set content if provided=0A=
      if (html) {=0A=
         this.innerdiv.innerHTML =3D html;=0A=
      }=0A=
=0A=
      this.div.style.left =3D x + "px";=0A=
      this.div.style.top =3D y + "px";=0A=
=0A=
      // Pop it=0A=
      utils.addClass(this.div, PopDiv.POPPED);=0A=
=0A=
   },=0A=
=0A=
   // Show it near element ("dir" indications direction)=0A=
   // Dimensions don't exist if element isn't shown.=0A=
   getXY: function() {=0A=
      var popped =3D utils.hasClass(this.div, PopDiv.POPPED);=0A=
=0A=
      if (!popped) {=0A=
         utils.addClass(this.div, PopDiv.POPPED);=0A=
      }=0A=
      var dim =3D utils.getXY(this.div);=0A=
      if (!popped) {=0A=
         utils.removeClass(this.div, PopDiv.POPPED);=0A=
      }=0A=
=0A=
      return dim;=0A=
   },=0A=
=0A=
   hide: function() {=0A=
      utils.removeClass(this.div, PopDiv.POPPED);=0A=
      if (this.popClass) {=0A=
         utils.removeClass(this.div, this.popClass);=0A=
         this.popClass =3D null;=0A=
         this.popnode =3D null;=0A=
      }=0A=
   },=0A=
=0A=
   // showBy shows a popper adjacent to another item.=0A=
   showBy: function(html, elem, dir) {=0A=
      var dx =3D 0;=0A=
      var dy =3D 0;=0A=
=0A=
      // Already popped over a particular popnode=0A=
      if (this.popnode && (this.popnode =3D=3D elem)) {=0A=
         return;=0A=
      }=0A=
      this.popnode =3D elem;=0A=
=0A=
      // Default direction=0A=
      if (typeof(dir) !=3D "number") {=0A=
         dir =3D PopDiv.POP_LEFT;=0A=
      }=0A=
=0A=
      // Set class "pop-<direction>": this determines margin=0A=
      this.popClass =3D PopDiv.POP_CLASSES[dir];=0A=
      //console.info("dir =3D " + dir +", popclass =3D " + =
this.popClass);=0A=
      utils.addClass(this.div, this.popClass);=0A=
      =0A=
      // Set text so that sizing is correct=0A=
      this.innerdiv.innerHTML =3D html;=0A=
=0A=
      // Get geometries of target and popper=0A=
      var thatdim =3D utils.getXY(elem);=0A=
      var popdim =3D this.getXY(this.div);=0A=
      //console.info("that =3D");=0A=
      //console.info(thatdim);=0A=
      //console.info("pop =3D ");=0A=
      //console.info(popdim);=0A=
=0A=
      // Calculate position=0A=
      switch (dir) {=0A=
      case PopDiv.POP_ABOVE: dy =3D -1*popdim.h;  break;=0A=
      case PopDiv.POP_LEFT:  dx =3D -1*popdim.w;  break;=0A=
      case PopDiv.POP_BELOW: dy =3D    thatdim.h; break;=0A=
      case PopDiv.POP_RIGHT: dx =3D    thatdim.w; break;=0A=
      default:=0A=
        throw "PopDiv: Invalid direction: " + dir;=0A=
      }=0A=
      //console.info("Popping at: [x=3D" + (thatdim.x + dx) + ", =
y=3D"+(thatdim.y + dy)+"]");=0A=
=0A=
      this.showAt(null, thatdim.x + dx, thatdim.y + dy);=0A=
   }=0A=
};=0A=
=0A=
=0A=
=0A=
// An ElementPopper shows a popup adjacent to a target element when the =
user mouses over the target.=0A=
// It handles browser events and manipulates a popper to show and hide =
detail data.=0A=
//=0A=
// When the mouse cursor enters the target, the content appears in the =
popper nearby.=0A=
//  When the mouse cursor leaves the target, the popper is hidden.=0A=
//=0A=
// This rather intense event handling code is necessary because =
mouseover and mouseout doesn't work=0A=
// as expected--mouseouts occur when moving over child nodes. The =
"relatedTarget" trick described=0A=
// by PPK (quirksmode.org) doesn't work either, because browsers =
sometimes drop events=0A=
// with related targets. So the only option is to capture =
document.onmousemove, and detect when=0A=
// the mouse either exits document.body, or moves over something that is =
neither the target nor=0A=
// one of its children.=0A=
//=0A=
// "popnode" is the target node=0A=
// "content" is either a string (the content to show), a DOM node (show =
its innerHTML), or a function.=0A=
// "config" is configuration; if it's just a number, then it's =
"direction" (PopDiv.POP_*); otherwise, it's a JS object with =0A=
//     attribute/value pairs=0A=
// "popperId" is the ID to assign to the popper; default is to make one =
up.=0A=
//=0A=
// When used with Ext.Ncbi, the popper should initialize itself on the =
page a few hundred=0A=
// ms after the page loads. IE apparently creates a race condition where =
popper and Ext.Ncbi initialization=0A=
// throw away the event handlers on which ElementPopper depends.=0A=
//=0A=
// So in that case, instead of new ElementPopper(a,b), you want to do:=0A=
//     setTimeout(function() {new ElementPopper(a,b);}), 500);=0A=
=0A=
//=0A=
var ElementPopper =3D function(popnode, content, config, popperId) {=0A=
=0A=
    var that =3D this;=0A=
=0A=
    // Inner functions=0A=
    // If target is popnode or one of its children, returns popnode; =
otherwise, null=0A=
    function getPopTarget(target) {=0A=
       while (target && !utils.hasClass(target, "popnode")) {=0A=
          target =3D target.parentNode;=0A=
       }=0A=
       return target;=0A=
    };=0A=
=0A=
    // Use inner functions here because removeEvent requires a reference =
to the=0A=
    // function object passed to addEvent, and we need to use closure to =
maintain scope.=0A=
=0A=
    // Show popper and start tracking mouse motions=0A=
    var handlePop =3D function(e) {=0A=
       var target =3D getPopTarget(utils.getTargetObj(e));=0A=
       =0A=
       if (target) {=0A=
	       // If Ajax URL is supplied, then fetch URL.=0A=
	       //   If AJAX URL is a function, the URL is the return value of =
the function=0A=
	       //   Otherwise AJAX URL must be a string=0A=
	       // When AJAX call succeeds, pop up the content. If there's a =
contentFunction, the content is=0A=
	       // the result of applying the contentFunction to the AJAX return =
data; otherwise, it's just=0A=
	       // the AJAX return data itself.=0A=
	       //=0A=
	       if (that.url) {=0A=
	          var url =3D (typeof(that.url) =3D=3D 'function') ? =
that.url(target) : that.url;=0A=
	=0A=
	          jQuery.ajax({=0A=
	             cache: false,=0A=
	             'url': url,=0A=
	             success: function(data, textStatus) {=0A=
	                doPop(target, data, textStatus);=0A=
	             }=0A=
	          });=0A=
	       } else {=0A=
	          doPop(target);=0A=
	       }=0A=
	       =0A=
	       // Set up to listen for mouse out=0A=
	       // This is the node currently being handled; it's cleared when=0A=
	       // the user mouses out.=0A=
	       that.handlingNode =3D target;=0A=
	    =0A=
           utils.removeEvent(target, "mousemove", handlePop);=0A=
           utils.addEvent(document.body, "mousemove", trackMouse);=0A=
           utils.addEvent(document.body, "mouseout", trackMouse);=0A=
       }=0A=
    }=0A=
    =0A=
    var doPop =3D function(target, data, status) {=0A=
       =0A=
       // Pop, shift listener to document body=0A=
       if (target) {=0A=
          that.timer =3D setTimeout(function() {=0A=
             // If we're still waiting for a pop, do the pop.=0A=
             // We're only waiting for a pop when handlingNode is set.=0A=
             if (target =3D=3D that.handlingNode) {=0A=
                that.thePopper.showBy(that.contentFunction(target, data, =
status), target, that.direction);=0A=
             } else {=0A=
               // Ignore the pop request, because we're no longer =
interested in this pop.=0A=
             }=0A=
             that.timer =3D null;             =0A=
          }, that.delay);=0A=
=0A=
       }=0A=
    };=0A=
=0A=
    // Unpop, shift listener back to popnode=0A=
    var trackMouse =3D function(e) {=0A=
       var target =3D getPopTarget(utils.getTargetObj(e));=0A=
=0A=
       // If mouse left document.body, or the target's not over the =
popnode, hide.=0A=
       if (e.type =3D=3D "mouseout" || target =3D=3D null) {=0A=
          // Clear pop timer if it hasn't popped=0A=
          if (that.timer) {=0A=
             window.clearTimeout(that.timer);=0A=
             that.timer =3D null;=0A=
          }=0A=
          that.thePopper.hide();=0A=
          that.handlingNode =3D null;=0A=
          =0A=
          utils.removeEvent(document.body, "mousemove", trackMouse);=0A=
          utils.removeEvent(document.body, "mouseout", trackMouse);=0A=
          utils.addEvent(popnode, "mousemove", handlePop);=0A=
       }=0A=
    };=0A=
=0A=
    // END inner functions=0A=
=0A=
    //=0A=
    // ctor main=0A=
    //=0A=
     =0A=
    // Support new signature ElementPopper(element, {config...}); in =
that case, "content" is a config object,=0A=
    // all other arguments are ignored, and config.content must be set =0A=
    if (typeof(content) =3D=3D 'object') {=0A=
       config =3D content;=0A=
       content =3D config.content;=0A=
    }=0A=
    =0A=
    // If a content URL is defined (AJAX URL string or function that =
returns URL), remember it=0A=
    if (config && config.url) {=0A=
       this.url =3D config.url;=0A=
    }=0A=
    =0A=
    // If content is a string, make it a function that returns the =
string.=0A=
    // If it's a dom node, make it a function that returns the dom =
node's innerHTML=0A=
    // If content is undefined, then assume data is coming from AJAX =
callback=0A=
    if (typeof(content) =3D=3D 'undefined') {=0A=
       this.contentFunction =3D function(target, data, status) { return =
data };=0A=
    } else if (typeof(content) =3D=3D 'string') {=0A=
       this.contentFunction =3D function() { return content; }=0A=
    } else if (content.innerHTML) {=0A=
       this.contentFunction =3D function() { return content.innerHTML; }=0A=
    } else if (typeof(content) =3D=3D 'function') {=0A=
       this.contentFunction =3D content; // Better be a function...=0A=
    }=0A=
    =0A=
    // Backward compatibility: "config" arg used to be "direction", so=0A=
    // interpret it as "direction" if config is a number.=0A=
    // Otherwise parse out config info=0A=
    if (typeof(config) =3D=3D=3D 'undefined') {=0A=
       config =3D {=0A=
          direction: PopDiv.POP_LEFT,=0A=
          delay: 0=0A=
       }=0A=
    }=0A=
    =0A=
    if (typeof(config) =3D=3D 'number') {=0A=
       this.direction =3D config;=0A=
    } else {=0A=
       this.direction =3D (typeof(config.direction) !=3D 'undefined') ? =
config.direction : PopDiv.POP_LEFT;=0A=
       this.delay =3D config.delay || 0;=0A=
    }=0A=
    this.popperId =3D config.popperId || popperId || ("popper" + =
(ElementPopper.popIndex++)); =0A=
=0A=
    // Create the popper=0A=
    this.thePopper =3D new PopDiv(this.popperId);=0A=
    var popdiv =3D $(this.popperId);=0A=
    =0A=
    // FIXME: Is this necessary? I think it's maybe wrong!=0A=
    if (popdiv) {=0A=
       utils.addClass(popdiv, "popnode");=0A=
    }=0A=
=0A=
    // Get popnode if it's an id string=0A=
    if (typeof(popnode) =3D=3D 'string') {=0A=
        popnode =3D $(popnode);=0A=
        if (!popnode) {=0A=
           throw "ElementPopper: id not found";=0A=
        }=0A=
    }=0A=
    this.popnode =3D popnode;=0A=
    this.timer =3D null; // Only exists when waiting for a pop=0A=
    =0A=
    // When the user mouses over target, pop it up=0A=
    utils.addEvent(popnode, "mousemove", handlePop);=0A=
}=0A=
=0A=
ElementPopper.popIndex =3D 0;=0A=
=0A=
ElementPopper.prototype =3D {=0A=
   setDelay: function(delay) {=0A=
      this.delay =3D delay;=0A=
   },=0A=
   getDelay: function() {=0A=
      return delay;=0A=
   }=0A=
};=0A=
=0A=
=0A=
=0A=
 utils.addEvent(window, "load", function(){     =0A=
     theDEP =3D new DeferredElementPopper("rapopper", { =0A=
          content: function(node) { =0A=
               var c =3D cssQuery(".hidden", node);               =0A=
               if (c && c.length > 0) {=0A=
                  var label =3D (c[0].textContent || c[0].innerText); =0A=
                  var cit =3D getCitation(node);=0A=
                  label =3D "<p id=3D'htbpoptext'>" + label + "</p>";=0A=
                  if (cit && cit > ""){ // avoid "undefined" value=0A=
                      label =3D label + "<p id=3D'htbpcit'>" + cit + =
"</p>";=0A=
                  } =0A=
                  return label;=0A=
               }                =0A=
          }, =0A=
          direction: PopDiv.POP_LEFT, =0A=
          popperId: "ppPopper", =0A=
          delay: 0 =0A=
     }); =0A=
});=0A=
=0A=
Portal.Portlet.HistoryDisplay =3D Portal.Portlet.extend({

	init: function(path, name, notifier) {
		console.info("Created History Ad...");
		this.base(path, name, notifier);   =20
	},
=09
	send: {
      'Cmd': null
      //'Recording': null
    },  =20
   =20
    receive: function(responseObject, userArgs) {  =0A=
         var cmd =3D userArgs.cmd;
         var rootNode =3D document.getElementById('HTDisplay'); =0A=
         var ul =3D document.getElementById('activity');=0A=
         var resp =3D responseObject.responseText;=0A=
             =0A=
         if (cmd =3D=3D 'HTOn') { =0A=
            rootNode.className =3D '';    // hide all msg and the turnOn =
link=0A=
            try {=0A=
                // Handle timeouts=0A=
                if (responseObject.status =3D=3D 408) { =0A=
                    rootNode.className =3D 'HTOn'; // so that the =
following msg will show up=0A=
                    rootNode.innerHTML =3D "<p class=3D'HTOn'>Your =
browsing activity is temporarily unavailable.</p>";=0A=
                    return;=0A=
                }=0A=
                   =0A=
                 // Looks like we got something...=0A=
                 resp =3D '(' + resp + ')';=0A=
                 var JSONobj =3D eval(resp);=0A=
                 =0A=
                 // Build new content (ul)=0A=
                 var newHTML =3D JSONobj.Activity;=0A=
                 var newContent =3D document.createElement('div');=0A=
                 newContent.innerHTML =3D newHTML;=0A=
                 var newUL =3D newContent.getElementsByTagName('ul')[0];=0A=
                 //alert(newHTML);=0A=
                 //alert(newContent.innerHTML);=0A=
                 //alert(newUL.innerHTML);=0A=
                 // Update content=0A=
                 rootNode.replaceChild(newUL, ul);=0A=
                 //XHR returns no activity (empty ul), e.g. activity =
cleared=0A=
                 if (newUL.className =3D=3D 'hide')                    =20
                     rootNode.className =3D 'HTOn';  // show "Your =
browsing activity is empty." message=0A=
                 =0A=
            }         =0A=
            catch (e) {=0A=
                //alert('error');=0A=
                rootNode.className =3D 'HTOn'; // so that the following =
msg will show up=0A=
                rootNode.innerHTML =3D "<p class=3D'HTOn'>Your browsing =
activity is temporarily unavailable.</p>";=0A=
           }=0A=
         }=0A=
         else if (cmd =3D=3D 'HTOff') {                         =0A=
             if (ul !=3D null) { =0A=
                 ul.className=3D'hide'; =0A=
                 ul.innerHTML =3D ''; // clear activity=0A=
             }=0A=
             rootNode.className =3D 'HTOff';    // make "Activity =
recording is turned off." and the turnOn link show up             =0A=
         }=0A=
         else if (cmd =3D=3D 'ClearHT') {     =0A=
             if ( rootNode.className =3D=3D '') { //                 =0A=
                 rootNode.className =3D 'HTOn';  // show "Your browsing =
activity is empty." message                                  =0A=
                 if (ul !=3D null) {=0A=
                     ul.className=3D'hide'; =0A=
                     ul.innerHTML =3D '';=0A=
                 }=0A=
             }            =0A=
         } =0A=
        =20
    },
   =20
	listen: {
	  'Cmd' : function(sMessage, oData, sSrc){
			console.info("Inside Cmd in HistoryDisplay: " + oData.cmd);
			this.setValue("Cmd", oData.cmd);
	  },	 =20
	=09
      "HistoryToggle<click>" : function(e, target, name){
         //alert(target.getAttribute("cmd"));
         this.send.Cmd({'cmd': target.getAttribute("cmd")});        =20
         console.info("Inside HistoryToggle in HistoryDisplay: " + =
target.getAttribute("cmd"));
        =20
         var site =3D document.forms[0]['p$st'].value;
         var cmd =3D  target.getAttribute("cmd");    =20
              =20
         // Issue asynchronous call to XHR service, callback is to =
update the portlet output
	     var resp =3D xmlHttpCall(site, this.path, cmd, {}, this.receive, =
{'cmd': target.getAttribute("cmd")}, this);
      },=20
     =20
      "HistoryOn<click>" : function(e, target, name){
         this.send.Cmd({'cmd': target.getAttribute("cmd")});
         =
//$PN('Pubmed_ResultsSearchController').getInput('RecordingHistory').valu=
e =3D 'yes';		=20
         console.info("Inside HistoryOn in HistoryDisplay: " + =
target.getAttribute("cmd"));
               =20
         var site =3D document.forms[0]['p$st'].value;  =20
	     var resp =3D xmlHttpCall(site, this.path, "HTOn", {}, =
this.receive, {'cmd': target.getAttribute("cmd")}, this);
	     //Portal.requestSubmit();
      },
     =20
      "ClearHistory<click>" : function(e, target, name){
         this.send.Cmd({'cmd': target.getAttribute("cmd")});=09
         var site =3D document.forms[0]['p$st'].value;  =20
	     var resp =3D xmlHttpCall(site, this.path, "ClearHT", {}, =
this.receive, {'cmd': target.getAttribute("cmd")}, this);	=20
      }
    }
});

function getCitation(node) {
   // var withHistorys =3D cssQuery("*[title]", node); does not work!!!  =
=20
   var withTitles =3D $AN('title', node, '*');
   //alert(withTitles.length + " " + withTitles[0].tagName + " " + =
withTitles[withTitles.length-1].tagName);
   //alert(withTitles[1].getAttribute('title'));
   if (withTitles.length > 0) {     =20
      var p =3D withTitles[0];
      var s =3D p.getAttribute("title");
      if (s && s > "") {=20
          //alert(s);
          p.setAttribute("title", "");
          node._title =3D s;
          for (var i =3D 0; i < withTitles.length; i++) {
              withTitles[i].removeAttribute("title");   =20
          }
      }
      /*else { //fixed in utils.js        =20
          s =3D node._title; // required for IE b/c of the bug in $AN
      }*/
   } else {
      s =3D node._title;
      //alert("old s "+ s);
   }
   return s;
}=0A=
// Requires: ElementPopper_JS, EventDispatcher_JS=0A=
=0A=
//=0A=
// A DeferredElementPopper creates an ElementPopper on any node with a =
given className the first time the=0A=
// node receives a mouseover event. It collects all of the poppers it =
creates (in an array), and=0A=
// remembers the dispatcher rule that invokes the initialization.=0A=
//=0A=
// "content" is the same as "content" for ElementPopper (see which)=0A=
//=0A=
DeferredElementPopper =3D function(className, config) {=0A=
   var that =3D this;=0A=
   this.config =3D config;=0A=
   this.poppers =3D [];=0A=
   this.dispatcher =3D new EventDispatcher("mouseover", className, =
function(event, udata, dispatcher) {=0A=
         if (!this._popup_init) {=0A=
            this._popup_init =3D 1;=0A=
            that.poppers[that.poppers.length] =3D new =
ElementPopper(this, that.config.content, that.config);=0A=
         }=0A=
    });=0A=
    =0A=
}=0A=
DeferredElementPopper.prototype =3D {=0A=
   debug: function() {=0A=
     var s =3D "";=0A=
     for (var i =3D 0; i < this.poppers.length; i++) {=0A=
        var z =3D this.poppers[i];=0A=
        s +=3D i + ": ";=0A=
        s +=3D z.popnode.innerText + "(" + =
z.popnode.getAttribute("title") +"); ";=0A=
     }=0A=
     alert(s);=0A=
   }=0A=
};=0A=
=0A=
=0A=
//=0A=
// A EventDispatcher does a popup on demand based on a classname on an =
element.=0A=
// Every mouse motion on the page notifies EventDispatcher that an event =
occurred.=0A=
// Any time an event occurs that has one of EventDispatcher's classnames =
on it,=0A=
// the handlers associated with that className are executed.=0A=
//=0A=
// EventDispatcher is designed for popups, but can be used for anything; =
the handler=0A=
// is a generic function:=0A=
//    handler(event, udata, dispatchrule)=0A=
// <event> is the platform event object; use utils.getTargetObj() to get =
its target=0A=
// <udata> is the udata that was passed when that handler was set up.=0A=
// <dispatchrule> is the EventDispatcher object; it is the dictionary=0A=
// {eventType, className, handler, udata} describing the popper behavior.=0A=
//=0A=
// dispatchrule.remove() [TBD] removes the rule from the event dispatcher=0A=
//=0A=
// When the callback occurs, <this> is the target element. =0A=
=0A=
EventDispatcher =3D function(eventType, className, handler, udata) {=0A=
=0A=
   // Start PP listening for this event=0A=
   if (!EventDispatcher.events) {=0A=
      EventDispatcher.events =3D {};=0A=
   }=0A=
=0A=
    // If first definition for this event type, listen for it=0A=
   if (!EventDispatcher.events[eventType]) {=0A=
      EventDispatcher.events[eventType] =3D {};=0A=
      utils.addEvent(document.body, eventType, =
EventDispatcher.handleEvent);=0A=
   }=0A=
   =0A=
   // Index of all class names managed by PP=0A=
   if (!EventDispatcher.classNames[className]) {=0A=
      EventDispatcher.classNames[className] =3D 1;=0A=
   }=0A=
=0A=
   // ev.classNames are the classes that listen for this event=0A=
   var ev =3D EventDispatcher.events[eventType];=0A=
   if (!ev.classNames) {=0A=
      ev.classNames =3D {};=0A=
   }=0A=
   =0A=
   // Add handler for this class=0A=
   if (!ev.classNames[className]) {=0A=
      ev.classNames[className] =3D [];=0A=
   }=0A=
   =0A=
   this.eventType =3D eventType;=0A=
   this.className =3D className;=0A=
   this.handler =3D handler;=0A=
   this.udata =3D udata || null;=0A=
   =0A=
   var c =3D ev.classNames[className];=0A=
   c[c.length] =3D this;=0A=
}=0A=
=0A=
EventDispatcher.events =3D {}=0A=
EventDispatcher.classNames =3D {}=0A=
=0A=
EventDispatcher.handleEvent =3D function(e) {=0A=
=0A=
   if (!EventDispatcher.events[e.type]) { return; } // TODO: Remove =
event listener here=0A=
   =0A=
   var t =3D utils.getTargetObj(e);=0A=
   if (t.className) {=0A=
      var theClasses =3D t.className.split(/\s+/);=0A=
      for (var c in theClasses) {=0A=
         if (EventDispatcher.classNames[theClasses[c]]) {=0A=
            EventDispatcher.dispatch(e, t, theClasses[c]);=0A=
         }=0A=
      }=0A=
   }=0A=
}=0A=
=0A=
EventDispatcher.dispatch =3D function(e, t, className) {=0A=
   if (!EventDispatcher.events[e.type]) return;=0A=
   var ev =3D EventDispatcher.events[e.type];=0A=
   if (!ev.classNames || !ev.classNames[className]) return;=0A=
   var poppers =3D ev.classNames[className];=0A=
   for (var i =3D 0; i < poppers.length; i++) {=0A=
      poppers[i].handleEvent(t, e);=0A=
   }=0A=
}=0A=
=0A=
EventDispatcher.prototype =3D {=0A=
   handleEvent: function(target, event) {=0A=
      this.handler.call(target, event, this.udata, this);=0A=
   }=0A=
}=0A=
=0A=
=0A=
=0A=
// PPMCRecentActivity=0A=
//=20
=0A=
Portal.Portlet.PPMCRecentActivity =3D =
Portal.Portlet.HistoryDisplay.extend({

    init: function(path, name, notifier) {
        console.info("Created PPMCRecentActivity");
        this.base(path, name, notifier);
    }
});=0A=
=0A=
=0A=
Portal.Portlet.Pubmed_Discovery_RA =3D =
Portal.Portlet.BriefLinkPageSection.extend({
   =20
    init: function(path, name, notifier) {
        console.info("Created Pubmed_Discovery_RA");
		this.base(path, name, notifier);
	},
=09
	SetPortletName: function(){
	    Portal.Portlet.BriefLinkPageSection.portletname =3D =
'Pubmed_Discovery_RA';
	}
});=0A=
Portal.Portlet.PPMCPubmedRA =3D =
Portal.Portlet.BriefLinkPageSection.extend({

    init: function(path, name, notifier) {=0A=
        console.info("Created PPMCPubmedRA");=0A=
        this.base(path, name, notifier);=0A=
    },=0A=
=0A=
    SetPortletName: function(){=0A=
        Portal.Portlet.BriefLinkPageSection.portletname =3D =
'PPMCPubmedRA';=0A=
    },=0A=
=0A=
    popNodeDelay: 500=0A=
});=0A=
// PPMCArticlePageJS=0A=
// This script does the following:=0A=
// - Positions reference portlets in the discovery column to the top of =
each paragraph;=0A=
// - Calculates how many references will fit;=0A=
// - Calculates whether it needs a "see all" link;=0A=
// - Adds mouseovers to links in paragraphs and reference portlet links=0A=
=0A=
// We use a self executing anonymous function in order to protect the =
global namespace from=0A=
// two top-level vars: '$j', and 'main'.=0A=
=0A=
( function() {=0A=
    var $j =3D jQuery;=0A=
=0A=
    // This main function runs when the DOM is ready to be queried; it =
gets attached to=0A=
    // the event down at the bottom of this module.=0A=
    var main =3D function() {=0A=
        //var start =3D new Date().getTime();=0A=
        var POPPER_DELAY =3D 500;=0A=
=0A=
        =
//////////////////////////////////////////////////////////////////////=0A=
        // Helper functions=0A=
=0A=
        =
//-------------------------------------------------------------------=0A=
        // This function determines if two rectangles intersect.=0A=
        // Two rectangles intersect if any point along their lines share =
a =0A=
        // point in space.  Because these portlets will be lined up =
vertically, =0A=
        // we only worry about one dimension.  The 'dims' parameter is =
an =0A=
        // object that contains top and bottom dimensions of the two =
rectangles.=0A=
=0A=
        var rectIntersect =3D function(dims) {=0A=
            // L1t is 'Line'1'Top' etc.=0A=
            var L1t =3D dims.L1t;=0A=
            var L1b =3D dims.L1b;=0A=
            var L2t =3D dims.L2t;=0A=
            var L2b =3D dims.L2b;=0A=
            return (=0A=
                (L2b > L1t && L2b < L1b) ||=0A=
                (L2t < L1b && L2t > L1t) ||=0A=
                (L2t > L1t && L2t < L1b) ||=0A=
                (L2t < L1t && L2b > L1b)=0A=
            );=0A=
        };=0A=
=0A=
        =
//-------------------------------------------------------------------=0A=
        // This function adds a popup to the links within the main =
article body.=0A=
        // It adds the popup to one element, and then queues itself up =
to add the=0A=
        // popup to the next element after a short delay.=0A=
=0A=
        var addPopper =3D function(elems, curr) {=0A=
            var currElem =3D elems[curr];=0A=
            var jqCurrElem =3D jQuery(currElem);=0A=
=0A=
            var rid =3D jqCurrElem.attr('rid');=0A=
            //console.info("rid is '" + rid + "'");=0A=
=0A=
            if (rid) {=0A=
                // Find the text for this popup from the element pointed =
to by =0A=
                // @rid.  This will be in the reference section of the =
article.=0A=
                // Some id's from the backend have special characters, =
so we=0A=
                // need to escape them, see PMC-6384 and =
http://tinyurl.com/2qfqgc=0A=
=0A=
                var ridEscaped =3D rid.replace(/:/g, "\\:")=0A=
                                    .replace(/\./g, "\\.")=0A=
                                    .replace(/=C2=B7/g, "\\=C2=B7");=0A=
                var refElem =3D jQuery('#' + ridEscaped);=0A=
                =0A=
                // Check to make sure that we found something=0A=
                if (refElem.length > 0) {=0A=
=0A=
                    var citeText =3D refElem.text();=0A=
                    //console.info("citeText is '" + citeText + "'");=0A=
=0A=
                    var popText =3D jQuery.trim(citeText);=0A=
                    //console.info("popText is '" + popText + "'");=0A=
    =0A=
                    // If the @href element of the current <a> tag =
(currElem)=0A=
                    // has a '#' portion, meaning that it is a link to =
the=0A=
                    // reference list in this same article, then =
indicate =0A=
                    // that in the popup text.=0A=
                    if (currElem.hash) {=0A=
                        popText +=3D ' [Reference List]';=0A=
                    }=0A=
    =0A=
                    new ElementPopper(currElem,=0A=
                        { content: popText, =0A=
                          direction: PopDiv.POP_ABOVE, =0A=
                          popperId: 'popper-para-links', =0A=
                          delay: POPPER_DELAY });=0A=
                }=0A=
            }=0A=
=0A=
            // Now arrange to have ourselves called for the next element =
that needs a popup,=0A=
            // after one millisecond.=0A=
            curr++;=0A=
            if (curr < elems.length) {=0A=
                var fnc =3D function(){=0A=
                    addPopper(elems, curr);=0A=
                }=0A=
                window.setTimeout(fnc, 1);=0A=
            }=0A=
        }; // end addPopper function=0A=
=0A=
        =
//-------------------------------------------------------------------=0A=
        // This function adds a popup to the links in the =
cited-reference blocks.=0A=
=0A=
        var addPopper2 =3D function(elems, curr) {=0A=
            var aDomElement =3D elems[curr];=0A=
            var aKids =3D aDomElement.childNodes;=0A=
            var aNumKids =3D aKids.length;=0A=
            //console.info("aDomElement is a " + aDomElement.toString() =
+ "; num kids is " + aNumKids);=0A=
=0A=
            // Generate the text that will go in the popup.=0A=
            // Get the text children of the <a> element.  This loop =
drops any element nodes;=0A=
            // in particular, the <span class=3D'flag'>Review</span> =
element, if present.=0A=
            var txt =3D "";=0A=
            for (var i =3D 0; i < aNumKids; ++i) {=0A=
                var kid =3D aKids[i];=0A=
                if (kid.nodeType !=3D 1) {=0A=
                    txt +=3D kid.nodeValue;=0A=
                }=0A=
            }=0A=
=0A=
            var l =3D jQuery(elems[curr]);=0A=
            txt +=3D "<br/>" + l.next('p')[0].innerHTML;=0A=
            new ElementPopper(l[0],=0A=
                { content: txt, direction: PopDiv.POP_LEFT, popperId: =
'popper-port-links', delay: POPPER_DELAY });=0A=
=0A=
            // Now arrange to have ourselves called for the next element =
that needs a popup,=0A=
            // after one millisecond.=0A=
            curr++;=0A=
            if (curr < elems.length) {=0A=
                var fnc =3D function(){=0A=
                    addPopper2(elems, curr);=0A=
                }=0A=
                window.setTimeout(fnc, 1);=0A=
            }=0A=
        }; // end addPopper2 function=0A=
=0A=
        // end helper function(s)=0A=
=0A=
        =
//////////////////////////////////////////////////////////////////////=0A=
        // In this space, we define and set variables that only need to =
be set once, not per paragraph.=0A=
=0A=
        // See if there any reference portlets in the page, by getting =
at least two reference=0A=
        // portlets. Later we will use these two portlets to make sure =
they do not interfere with=0A=
        // existing portlets in the column. It is unlikely that a third =
portlet would interfere=0A=
        // with anything, so let's make this as efficient as possible=0A=
        var portNodes =3D =
$j('#ArticleRefDocsums').find('div.cited-ref:lt(2)');=0A=
=0A=
        // If there is at least one portlet in the page, we need to get =
some initial information,=0A=
        // like sample portlet elements' sizes, information about =
existing portlet in page etc.=0A=
        var portsInPage =3D false;=0A=
        if (portNodes.length > 0) { // if the page has ANY reference =
portlets=0A=
            // remember that there is something to do=0A=
            var portsInPage =3D true;=0A=
            var firstPortN =3D $j(portNodes[0]);=0A=
=0A=
            // Max shown per page is already decided by xslt that =
generates portlet server-side,=0A=
            // and should be the number of <li>s per portlet=0A=
            var MAXREFS =3D 5;=0A=
=0A=
            // get dimensions of elements inside sample portlet, account =
for padding by using outerHeight=0A=
            var lis =3D firstPortN.find('li.ovfl');=0A=
            var firstRefH =3D $j(lis[0]).outerHeight(true);=0A=
            var firstSeeAllH =3D =
firstPortN.find('a.all-articles').outerHeight(true);=0A=
            var firstUl =3D firstPortN.find('ul');=0A=
            // the ul might have padding or margins applied=0A=
            var ulPaddingMargin =3D parseInt(firstUl.css('padding-top')) =
+=0A=
                                  =
parseInt(firstUl.css('padding-bottom')) +=0A=
                                  parseInt(firstUl.css('margin-top')) +=0A=
                                  parseInt(firstUl.css('margin-bottom'));=0A=
            // same goes for the actual portlet div=0A=
            var portNPadding =3D parseInt(firstPortN.css('padding-top')) =
+=0A=
                               =
parseInt(firstPortN.css('padding-bottom'));=0A=
=0A=
            // The horizontal position of all cited ref blocks should =
line up with recent history=0A=
            // portlet, which should be above any reference portlets=0A=
            var leftPos =3D $j('#recent-activity').position().left;=0A=
            //console.info('Got leftPos of all portlets: ' + leftPos);=0A=
=0A=
            // Get positions of any existing portlets in side column, so =
we can later hide any=0A=
            // reference portlets that take up same space=0A=
            var sideBarCell =3D $j('td.sidebar-cell')[0];=0A=
            var portRanges =3D {};=0A=
            var ports =3D jQuery('div.port', sideBarCell);=0A=
            for (var i =3D 0; i < ports.length; i++) {=0A=
                var p =3D jQuery(ports[i]);=0A=
                var pos =3D p.position();=0A=
                var t =3D pos.top;=0A=
                portRanges[i] =3D {=0A=
                    't': t,=0A=
                    'b': t + p.height() // bottom is top + height of =
portlet=0A=
                };=0A=
            }=0A=
=0A=
        } // end if portNodes in doc=0A=
=0A=
        // Now, for each paragraph, get associated portlet and position =
appropriately, we still need=0A=
        // to iterate through each paragraph even if there are no =
reference links in entire document.=0A=
        // We are doing some weird stuff here.  We have to do iterate =
through each paragraph, but=0A=
        // for performance reasons, on large pages we need to pause a ms =
between iterations. So:=0A=
        // 1) we get all "paragraph" elements=0A=
        // 2) we execute the "loopdloop" function, which every ms passes =
all the paras to paraLoop=0A=
        // 3) paraLoop does all the positioning of the reference portlets=0A=
        // 4) When loopdloop is done, it calls hideInterfering, which =
hides any portlets =0A=
        //    that overlap existing ones=0A=
=0A=
        // First make a hash of all of the CRBs indexed by their @rid =
attributes.=0A=
        var crbs =3D $j('div.cited-ref');=0A=
        var numCrbs =3D crbs.length;=0A=
        //console.info("Number of crbs is " + numCrbs);=0A=
=0A=
        var crbsByRid =3D {};=0A=
        for (var i =3D 0; i < numCrbs; ++i) {=0A=
            var crb =3D crbs[i];=0A=
            //console.info("crb is " + crb.toString());=0A=
            var jcrb =3D $j(crb);=0A=
            //console.info("jcrb is " + jcrb.toString());=0A=
            var rid =3D jcrb.attr('rid');=0A=
            //console.info("@rid is '" + rid + "'");=0A=
            crbsByRid[rid] =3D jcrb;=0A=
        }=0A=
=0A=
        //var paraLoop =3D function(crb) {=0A=
        //    var jcrb =3D $j(crb);=0A=
        //    var paraRid =3D jcrb.attr('rid');=0A=
        //    var p =3D $j('#' + paraRid);=0A=
        var paraLoop =3D function(para) {=0A=
            //console.group('para');=0A=
            var p =3D $j(para);=0A=
            //console.info(p[0]);=0A=
=0A=
            // Get the @id attribute, and from that, look up the CRB.=0A=
            var paraId =3D p.attr('id');=0A=
            //console.info("paragraph @id is " + paraId);=0A=
            var jcrb =3D crbsByRid[paraId];=0A=
=0A=
            if (jcrb) {=0A=
                //console.info("jcrb is " + jcrb.toString());=0A=
=0A=
                // if a 'paragraph' has an associated portlet, it will =
have a 'cite' attribute on it=0A=
                // CFM:  taking this check out, the @cite attribute is =
going away, see PMC-5595:=0A=
                //var citeAttr =3D p.attr('cite');=0A=
                //if (citeAttr) {=0A=
=0A=
                // Get the number of unique references per paragraph.  =
This could be more than the=0A=
                // max shown in portlet.=0A=
                // CFM:  Now, getting this info from the crb, not the =
paragraph, because the @pmids=0A=
                // attribute will be going away from the paragraph, see =
PMC-5595.=0A=
//              var numRefs =3D p.attr('pmids').split(' ').length;=0A=
                var numRefs =3D jcrb.attr('pmids').split(' ').length;=0A=
                //console.info("numRefs is " + numRefs);=0A=
=0A=
                // get paragraph dims. We need these to position portlet =
to paragraph=0A=
                var pPos =3D p.position();=0A=
                var pPosX =3D pPos.left;=0A=
=0A=
                 // needs to be a number because we will add/subtract =
margin=0A=
                var pPosY =3D parseInt(pPos.top);=0A=
                var pPosH =3D p.height();=0A=
=0A=
                // Get paragraph top and bottom margins.  We need to =
account for these when calculating =0A=
                // reference portlet placement. Margin is not accounted =
for when getting an element's =0A=
                // height, but it does affect appearance of element's =
dimensions when there is no border.=0A=
                var pMarT =3D parseInt(p.css('marginTop'));=0A=
                var pMarB =3D parseInt(p.css('marginBottom'));=0A=
=0A=
                // Get associated portlet from 'cited' attribute, we can =
now safely overwrite portN =0A=
                // variable from beginning of script.=0A=
                // CFM:  change from using 'portN' to using the variable =
'jcrb', from above.=0A=
                //var portN =3D $j('#' + citeAttr);=0A=
                //var refNodes =3D portN.find('li');=0A=
                var refNodes =3D jcrb.find('li');=0A=
                var numRefsInP =3D refNodes.length;=0A=
                //console.info("numRefsInP is " + numRefsInP);=0A=
=0A=
                // Set styles on portlet node to align to paragraph=0A=
                // We set portlet node to position: absolute in the css =
file, because that is more=0A=
                // efficient than setting it in JavaScript, and that =
property does not change.=0A=
//              portN.css( {=0A=
                jcrb.css( {=0A=
                    'top': pPosY + pMarT + 2 + 'px', // Add an xtra 2px, =
otherwise it appears too low=0A=
                    'left': leftPos + 'px'=0A=
                });=0A=
=0A=
                // Calculate how many refs will fit per paragraph and if =
we need the =0A=
                // 'See more articles cited in this paragraph' link. =0A=
                var refsFit =3D null;=0A=
                var needAllLink =3D false;=0A=
                if (numRefs > MAXREFS || numRefs * firstRefH >=3D pPosH =
- pMarB) {=0A=
                    //console.info('more links than allowed, OR links =
allowed are higher than paragraph height');=0A=
                    refsFit =3D Math.floor( pPosH / (firstRefH + =
firstSeeAllH) );=0A=
                    needAllLink =3D true;=0A=
                    if (refsFit > MAXREFS) {=0A=
                        refsFit =3D MAXREFS;=0A=
                    }=0A=
                    //console.info('refsFit: ' + refsFit);=0A=
                } else {=0A=
                    //console.info('all refs in para fit');=0A=
                    refsFit =3D numRefs;=0A=
                    //console.info('refsFit: ' + refsFit);=0A=
                }=0A=
=0A=
                var aSeeArticles =3D jcrb.find('a.all-articles');=0A=
                if (!needAllLink) {=0A=
                    aSeeArticles.hide();=0A=
                }=0A=
                else if (refsFit =3D=3D 0) {=0A=
                    // If the visible number of links is zero, then =
change the text from=0A=
                    // "See more articles ..." to just "See articles =
...".=0A=
                    // See PMC-6076.=0A=
                    var seeMoreT =3D aSeeArticles.text();=0A=
                    aSeeArticles.text( seeMoreT.replace(/more /, "") );=0A=
                    // After all, we decided that we'd rather just not =
see it:=0A=
                    jcrb.hide();=0A=
                }=0A=
=0A=
                //console.info('need to take off refs:');=0A=
                // how many references (lis) from the portlet do we need =
to hide?=0A=
                var takeOff =3D numRefsInP - refsFit;=0A=
                //console.info(takeOff);=0A=
=0A=
                if ( takeOff > 0 ) {=0A=
                    // we need to take off some nodes, loop through li =
nodes in reverse order and hide them=0A=
                    for (var i =3D numRefsInP - 1; i >=3D refsFit; i--) {=0A=
                        $j(refNodes[i]).hide();=0A=
                    }=0A=
                } // end if we need to take off nodes=0A=
=0A=
                // we now need to add mouseovers on reference links that =
are not hidden. We get the text=0A=
                // from the title attribute of the link=0A=
                //var elems2 =3D portN.find('a.popnode:visible');=0A=
                var elems2 =3D jcrb.find('a.popnode:visible');=0A=
                //console.info(elems2);=0A=
=0A=
                if (elems2.length > 0) {=0A=
                    addPopper2(elems2, 0);=0A=
                }=0A=
=0A=
            } // end if jcrb=0A=
=0A=
            // Now, we are back touching on each paragraph, regardless =
whether there =0A=
            // are links with pmids or not we need to get all the =
reference links per =0A=
            // paragraph and do the mouseover which links to reference =
section at the=0A=
            // bottom of the page.  This is a little Eric trick for =
performance. We get =0A=
            // the links in the paragraph and create our own custom loop =
and setTimeout =0A=
            // for each link.  This allows "multithreading" and doesn't =
lock up the =0A=
            // browser.=0A=
            var elems =3D p.find('a.cite-reflink');=0A=
            if (elems.length > 0) {=0A=
                addPopper(elems, 0);=0A=
            }=0A=
=0A=
            //console.groupEnd('para');=0A=
        }; // end paraloop function=0A=
=0A=
        var hideInterfering =3D function(){=0A=
            // At this point all reference divs are positioned in =
document. We now check that =0A=
            // the first two reference portlets (which were already =
queried for in beginning =0A=
            // of the script don't interfere with any other portlets =
that may be at the top of =0A=
            // column.  Loop thru them and compare them against what =
exists in portRanges=0A=
            if (portsInPage =3D=3D=3D true) {=0A=
                portNodes.each( function() {=0A=
                    var citedRefJ =3D $j(this);=0A=
                    // We refer to portRanges, which is an obj in which =
we stored dims of existing=0A=
                    // portlets before positioning reference portlets=0A=
                    for (var i in portRanges){=0A=
                        var range =3D portRanges[i];=0A=
=0A=
                        var existingT =3D range.t;=0A=
                        var existingB =3D range.b;=0A=
=0A=
                        var citedRefT =3D citedRefJ.position().top;=0A=
=0A=
                        // This is the parameter to the rectIntersect =
helper func defined in the beginning=0A=
                        // of the script.  It contains top and bottom =
dims for two rectanges.  The=0A=
                        // rectIntersect returns true if they intersect.=0A=
                        // PMC-6175:  add a little buffer of ten pixels.=0A=
                        var dims =3D {=0A=
                            'L1t': existingT, // we are comparing =
against the existing portlet=0A=
                            'L1b': existingB + 10,  // little buffer to =
be sure.=0A=
                            'L2t': citedRefT,=0A=
                            'L2b': citedRefT + citedRefJ.height()=0A=
                        }=0A=
                        //console.info('L1t: ' + existingT);=0A=
                        //console.info('L1b: ' + existingB);=0A=
                        //console.info('L2t: ' + citedRefT);=0A=
                        //console.info('L2b: ' + (citedRefT + =
citedRefJ.height()));=0A=
=0A=
                        if ( rectIntersect(dims) =3D=3D=3D true) {=0A=
                            //console.info('interferes');=0A=
                            citedRefJ.css('display', 'none');=0A=
                        }=0A=
                    }=0A=
                }); // end iterating through first two existing portlets=0A=
            }=0A=
        }=0A=
=0A=
        // Get the list of all paragraphs in the document.=0A=
        var allParas =3D $j('div.p');=0A=
        var numParas =3D allParas.length;=0A=
=0A=
        var loopdloop =3D function(curr){=0A=
          paraLoop(allParas[curr++]);=0A=
          if (curr >=3D numParas) {=0A=
            hideInterfering();=0A=
          } else {=0A=
            setTimeout( function(){ loopdloop(curr); }, 1);=0A=
          }=0A=
        }=0A=
        //document.title =3D new Date().getTime() - start.toString() + =
'ms';=0A=
        loopdloop(0);=0A=
    };=0A=
=0A=
=0A=
    // We use window.load for Safari, not ready, because in Safari, we =
cannot be =0A=
    // sure all CSS has loaded, and we cannot be sure=0A=
    // dims of images are accounted for when getting top positions of =
paragraphs=0A=
    if ( $j.browser.safari) {=0A=
        $j(window).load(main);=0A=
    } else {=0A=
        $j(main);=0A=
    }=0A=
=0A=
})();=0A=
=0A=

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