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Subject: =?Windows-1252?Q?Worse_Outcome_in_Primary_Glioblastoma_Multiforme_With_Co?=
	=?Windows-1252?Q?ncurrent_Epidermal_Growth_Factor_Receptor_and_p53_Altera?=
	=?Windows-1252?Q?tion_=97_American_Journal_of_Clinical_Pathology?=
Date: Tue, 10 Feb 2009 21:19:51 +0100
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Primary Glioblastoma Multiforme With Concurrent Epidermal Growth Factor =
Receptor and p53 Alteration =E2=80=94 American Journal of Clinical =
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content=3D"Worse Outcome in Primary Glioblastoma Multiforme With =
Concurrent Epidermal Growth Factor Receptor and p53 Alteration">
<META name=3DDC.Identifier content=3D10.1309/AJCP64YBDVCTIRWV>
<META name=3DDC.Date content=3D2009-02-01>
<META name=3DDC.Publisher content=3D"American Society for Clinical =
Pathology">
<META name=3DDC.Contributor content=3D"Yolanda Ruano">
<META name=3DDC.Contributor content=3D"Teresa Ribalta">
<META name=3DDC.Contributor content=3D"=C3=81ngel Rodr=C3=ADguez de =
Lope">
<META name=3DDC.Contributor content=3D"Yolanda Campos-Mart=C3=ADn">
<META name=3DDC.Contributor content=3D"Concepci=C3=B3n Fia=C3=B1o">
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<META name=3DDC.Contributor content=3D"Jos=C3=A9-Luis =
Hern=C3=A1ndez-Moneo">
<META name=3DDC.Contributor content=3D"Manuela Mollejo">
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content=3D"Ruano, Yolanda; Ribalta, Teresa; de Lope, =C3=81ngel =
Rodr=C3=ADguez; Campos-Mart=C3=ADn, Yolanda; Fia=C3=B1o, =
Concepci=C3=B3n; P=C3=A9rez-Mag=C3=A1n, Elisa; Hern=C3=A1ndez-Moneo, =
Jos=C3=A9-Luis; Mollejo, Manuela; Mel=C3=A9ndez, B=C3=A1rbara">
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Concurrent Epidermal Growth Factor Receptor and p53 Alteration">
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<H1 id=3Darticle-title-1>Worse Outcome in Primary Glioblastoma =
Multiforme With=20
Concurrent Epidermal Growth Factor Receptor and p53 Alteration</H1>
<DIV class=3Dcontributors>
<OL id=3Dcontrib-group-1 class=3Dcontributor-list>
  <LI id=3Dcontrib-1><SPAN class=3Dname><A class=3Dname-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?author1=3DYolanda+Ruano&amp;s=
ortspec=3Ddate&amp;submit=3DSubmit">Yolanda=20
  Ruano</A></SPAN><A id=3Dxref-aff-1-1 class=3Dxref-aff=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#aff-1">1</A>,=
=20
  <LI id=3Dcontrib-2><SPAN class=3Dname><A class=3Dname-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?author1=3DTeresa+Ribalta&amp;=
sortspec=3Ddate&amp;submit=3DSubmit">Teresa=20
  Ribalta</A></SPAN><SPAN class=3Dcontrib-degrees>, PhD, MD</SPAN><A=20
  id=3Dxref-aff-4-1 class=3Dxref-aff=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#aff-4">2</A>,=
=20
  <LI id=3Dcontrib-3><SPAN class=3Dname><A class=3Dname-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?author1=3D%C3%81ngel+Rodr%C3%=
ADguez+de+Lope&amp;sortspec=3Ddate&amp;submit=3DSubmit">=C3=81ngel=20
  Rodr=C3=ADguez de Lope</A></SPAN><SPAN class=3Dcontrib-degrees>, PhD, =
MD</SPAN><A=20
  id=3Dxref-aff-2-1 class=3Dxref-aff=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#aff-2">3</A>,=
=20
  <LI id=3Dcontrib-4><SPAN class=3Dname><A class=3Dname-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?author1=3DYolanda+Campos-Mart=
%C3%ADn&amp;sortspec=3Ddate&amp;submit=3DSubmit">Yolanda=20
  Campos-Mart=C3=ADn</A></SPAN><SPAN class=3Dcontrib-degrees>, =
PhD</SPAN><A=20
  id=3Dxref-aff-1-2 class=3Dxref-aff=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#aff-1">1</A>,=
=20
  <LI id=3Dcontrib-5><SPAN class=3Dname><A class=3Dname-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?author1=3DConcepci%C3%B3n+Fia=
%C3%B1o&amp;sortspec=3Ddate&amp;submit=3DSubmit">Concepci=C3=B3n=20
  Fia=C3=B1o</A></SPAN><SPAN class=3Dcontrib-degrees>, MD</SPAN><A =
id=3Dxref-aff-5-1=20
  class=3Dxref-aff=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#aff-5">4</A>,=
=20
  <LI id=3Dcontrib-6><SPAN class=3Dname><A class=3Dname-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?author1=3DElisa+P%C3%A9rez-Ma=
g%C3%A1n&amp;sortspec=3Ddate&amp;submit=3DSubmit">Elisa=20
  P=C3=A9rez-Mag=C3=A1n</A></SPAN><A id=3Dxref-aff-1-3 class=3Dxref-aff=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#aff-1">1</A>,=
=20
  <LI id=3Dcontrib-7><SPAN class=3Dname><A class=3Dname-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?author1=3DJos%C3%A9-Luis+Hern=
%C3%A1ndez-Moneo&amp;sortspec=3Ddate&amp;submit=3DSubmit">Jos=C3=A9-Luis =

  Hern=C3=A1ndez-Moneo</A></SPAN><SPAN class=3Dcontrib-degrees>, =
MD</SPAN><A=20
  id=3Dxref-aff-2-2 class=3Dxref-aff=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#aff-2">3</A>,=
=20
  <LI id=3Dcontrib-8><SPAN class=3Dname><A class=3Dname-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?author1=3DManuela+Mollejo&amp=
;sortspec=3Ddate&amp;submit=3DSubmit">Manuela=20
  Mollejo</A></SPAN><SPAN class=3Dcontrib-degrees>, PhD, MD</SPAN><A=20
  id=3Dxref-aff-1-4 class=3Dxref-aff=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#aff-1">1</A><=
A=20
  id=3Dxref-aff-3-1 class=3Dxref-aff=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#aff-3">5</A> =
and=20
  <LI id=3Dcontrib-9><SPAN class=3Dname><A class=3Dname-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?author1=3DB%C3%A1rbara+Mel%C3=
%A9ndez&amp;sortspec=3Ddate&amp;submit=3DSubmit">B=C3=A1rbara=20
  Mel=C3=A9ndez</A></SPAN><SPAN class=3Dcontrib-degrees>, PhD</SPAN><A =
id=3Dxref-aff-1-5=20
  class=3Dxref-aff=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#aff-1">1</A> =

</LI></OL>
<OL class=3Daffiliation-list>
  <LI class=3Daff><A id=3Daff-1 name=3Daff-1></A>
  <ADDRESS><SUP>1</SUP>From the Molecular Pathology Research Unit and =
</ADDRESS>
  <LI class=3Daff><A id=3Daff-2 name=3Daff-2></A>
  <ADDRESS><SUP>3</SUP>the Departments of Neurosurgery and </ADDRESS>
  <LI class=3Daff><A id=3Daff-3 name=3Daff-3></A>
  <ADDRESS><SUP>5</SUP>Pathology, Virgen de la Salud Hospital, Toledo, =
Spain;=20
  </ADDRESS>
  <LI class=3Daff><A id=3Daff-4 name=3Daff-4></A>
  <ADDRESS><SUP>2</SUP>Cl=C3=ADnic Hospital, Barcelona University, =
August Pi i Sunyer=20
  Biomedical Research Unit, Barcelona, Spain; and </ADDRESS>
  <LI class=3Daff><A id=3Daff-5 name=3Daff-5></A>
  <ADDRESS><SUP>4</SUP>Department of Pathology, Xeral-Cies Hospital =
Complex,=20
  Vigo, Spain </ADDRESS></LI></OL></DIV>
<DIV id=3Dabstract-1 class=3D"section abstract">
<DIV class=3Dsection-nav>
<DIV class=3Dnav-placeholder></DIV><A class=3Dnext-section-link=20
title=3D"Materials and Methods"=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#sec-1"><SPAN>=
Next=20
Section</SPAN></A></DIV>
<H2>Abstract</H2>
<P id=3Dp-1>Primary glioblastoma multiforme (GBM), in contrast with =
secondary GBM,=20
has been associated with the presence of <EM>EGFR</EM> amplification and =
absence=20
of <EM>p53</EM> mutation. In this study, we analyzed relevant molecular =
and=20
clinical variables in 194 primary GBMs and tested them for survival =
analysis.=20
Although most of the tumors showed a mutually exclusive pattern, =
concurrent=20
alterations of EGFR and p53 were detected. Survival analysis of =
<EM>CDK4</EM>=20
amplification revealed a highly significant association with a worse =
clinical=20
outcome (<EM>P</EM> =3D .01), whereas MDM2, CDK6, PTEN, and p21 were not =

associated with patient survival. Multivariate analysis including the=20
significant clinical and molecular variables revealed <EM>CDK4</EM>=20
amplification, age, and radiotherapy to be markers with independent =
prognostic=20
value. In addition, the primary GBM tumors showing simultaneous EGFR and =
p53=20
alterations were significantly associated with worse survival =
(<EM>P</EM> &lt;=20
.01). These results highlight the prognostic value of <EM>CDK4</EM>=20
amplification and of simultaneous EGFR-p53 alterations in the clinical =
outcome=20
of patients with primary GBM. </P></DIV><SPAN =
class=3Dkwd-group-title>Key=20
Words:</SPAN>
<UL class=3Dkwd-group>
  <LI class=3Dkwd><SPAN><A class=3Dkwd-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?fulltext=3DGlioblastoma&amp;s=
ortspec=3Ddate&amp;submit=3DSubmit&amp;andorexactfulltext=3Dphrase">Gliob=
lastoma</A></SPAN>=20

  <LI class=3Dkwd><SPAN><A class=3Dkwd-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?fulltext=3DEGFR&amp;sortspec=3D=
date&amp;submit=3DSubmit&amp;andorexactfulltext=3Dphrase">EGFR</A></SPAN>=
=20

  <LI class=3Dkwd><SPAN><A class=3Dkwd-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?fulltext=3Dp53&amp;sortspec=3D=
date&amp;submit=3DSubmit&amp;andorexactfulltext=3Dphrase">p53</A></SPAN> =


  <LI class=3Dkwd><SPAN><A class=3Dkwd-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?fulltext=3DCDK4&amp;sortspec=3D=
date&amp;submit=3DSubmit&amp;andorexactfulltext=3Dphrase">CDK4</A></SPAN>=
=20

  <LI class=3Dkwd><SPAN><A class=3Dkwd-search=20
  =
href=3D"http://ajcp.ascpjournals.org/search?fulltext=3DSurvival&amp;sorts=
pec=3Ddate&amp;submit=3DSubmit&amp;andorexactfulltext=3Dphrase">Survival<=
/A></SPAN>=20
  </LI></UL>
<P id=3Dp-2>Glioblastoma multiforme (GBM) is the most frequent primary =
brain tumor=20
in adults. GBMs are defined by the histopathologic features of cellular =
atypia,=20
mitotic figures, necrotic foci with peripheral cellular =
pseudopalisading, and=20
microvascular hyperplasia, which distinguish them from lower-grade =
astrocytic=20
tumors.<SUP><A id=3Dxref-ref-1-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-1">1</A><=
/SUP></P>
<P id=3Dp-3>Scherer<SUP><A id=3Dxref-ref-2-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-2">2</A><=
/SUP>=20
first classified GBMs into primary and secondary tumors in 1940. Primary =
GBMs=20
occur de novo without a recognizable precursor lesion. The clinical =
history of=20
the disease is usually short, with a median survival of approximately 1 =
year=20
following diagnosis. In contrast, secondary GBMs develop slowly and =
arise from=20
preexisting, lower-grade astrocytomas.<SUP><A id=3Dxref-ref-2-2 =
class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-2">2</A>=E2=
=80=93<A=20
id=3Dxref-ref-4-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-4">4</A><=
/SUP></P>
<P id=3Dp-4>Although these GBM subtypes are histologically =
indistinguishable,=20
molecular genetic analyses suggest that there are at least 2 distinct =
pathways=20
contributing to the tumorigenesis of GBM. Primary GBMs frequently show=20
amplification and/or overexpression of the epidermal growth factor =
receptor=20
(EGFR), <EM>PTEN</EM> mutations, and partial or complete loss of =
chromosome 10.=20
Otherwise, secondary GBMs generally feature mutations of the =
<EM>p53</EM> gene,=20
over-expression of platelet-derived growth factor receptors, and loss of =

heterozygosity at 17p, 19q, and 10q. In addition, they rarely =
overexpress=20
EGFR.<SUP><A id=3Dxref-ref-5-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-5">5</A>=E2=
=80=93<A=20
id=3Dxref-ref-8-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-8">8</A><=
/SUP>=20
Therefore, EGFR amplification and/or overexpression and <EM>p53</EM> =
mutation=20
are 2 genetic events that seem to molecularly differentiate these =
clinical=20
subtypes of GBM. Nevertheless, while the association of these 2 =
hallmarks has=20
been considered to be almost mutually exclusive, fluorescence in situ=20
hybridization (FISH) and reverse transcription=E2=80=93polymerase chain =
reaction=20
expression studies have shown coexistence of <EM>p53</EM> mutation and=20
<EM>EGFR</EM> amplification in subsets of primary GBM tumors.<SUP><A=20
id=3Dxref-ref-9-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-9">9</A>=E2=
=80=93<A=20
id=3Dxref-ref-11-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-11">11</A=
></SUP>=20
To our knowledge, the prognostic significance of these concurrent =
alterations=20
has not been studied. </P>
<P id=3Dp-5>On the other hand, frequent alterations of genes coding for =
proteins=20
involved in G<SUB>1</SUB>-S cell-cycle transition control have been =
reported in=20
primary and secondary GBMs. This is the case for the human homologue of =
the=20
mouse double minute 2 (<EM>MDM2</EM>), cyclin-dependent kinases 4 and 6=20
(<EM>CDK4</EM> and <EM>CDK6</EM>), and the inhibitor of cyclin-dependent =
kinases=20
<EM>p21.</EM><SUP><A id=3Dxref-ref-12-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-12">12</A=
>=E2=80=93<A=20
id=3Dxref-ref-16-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-16">16</A=
></SUP>=20
While coamplification of <EM>MDM2</EM> and <EM>CDK4</EM> has been =
observed in=20
gliomas, MDM2 amplification and/or over-expression is more frequent in =
primary=20
than in secondary GBMs. Conversely, <EM>CDK4</EM> amplification is =
considered a=20
late event during astrocytoma progression and is frequently found in =
secondary=20
GBMs.<SUP><A id=3Dxref-ref-17-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-17">17</A=
>,<A=20
id=3Dxref-ref-18-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-18">18</A=
></SUP></P>
<P id=3Dp-6>This study was undertaken to explore the prognostic =
relevance of EGFR=20
and p53 and other growth-control molecules such as MDM2, CDK4, CDK6, =
PTEN, and=20
p21 in a large series of primary GBMs. </P>
<DIV id=3Dsec-1 class=3D"section materials-methods">
<DIV class=3Dsection-nav><A class=3Dprev-section-link title=3DAbstract=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#abstract-1"><=
SPAN>Previous=20
Section</SPAN></A><A class=3Dnext-section-link title=3DResults=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#sec-6"><SPAN>=
Next=20
Section</SPAN></A></DIV>
<H2>Materials and Methods</H2>
<H3>Cases</H3>
<P id=3Dp-7 class=3Dh-lead>The study involved 194 GBM samples obtained =
between 1999=20
and 2008 from 3 hospitals in Spain: Virgen de la Salud (Toledo), Clinic=20
(Barcelona), and Xeral-Cies (Vigo). All cases were reviewed by 3 of us =
(T.R.,=20
C.F., and M.M.) to verify tumor viability and to confirm the diagnosis =
according=20
to the 2007 World Health Organization diagnostic criteria.<SUP><A=20
id=3Dxref-ref-17-2 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-17">17</A=
></SUP>=20
The selected patients did not have a history of brain tumor, so the =
clinical and=20
genetic profiles of the cases are characteristic of a primary (de novo) =
GBM=20
subtype. The patients received conventional therapy consisting of =
surgical=20
resection of as much of the tumor as clinically and technically =
possible,=20
followed by radiotherapy and/or chemotherapy. The median age of the =
patients at=20
the time of diagnosis was 59.5 years (range, 16=E2=80=9381 years), the =
median overall=20
survival was 10 months (range, 1=E2=80=9372 months), and the median =
postsurgical=20
Karnofsky performance score (P-KPS) was 80 (range, 50=E2=80=93100). =
Clinical information=20
is summarized in <A id=3Dxref-table-wrap-1-1 class=3Dxref-table=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#T1">Table =
1</A>. The=20
investigation was performed following the approval of the institutional =
review=20
board. </P>
<DIV id=3DT1 class=3Dtable>
<DIV class=3Dtable-inline>
<DIV class=3Dcallout><SPAN>View this table:</SPAN>
<UL class=3Dcallout-links>
  <LI><A=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/T1.expansion.html"=
>In=20
  this window</A>=20
  <LI><A class=3Din-nw=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/T1.expansion.html"=
>In a=20
  new window</A> </LI></UL></DIV></DIV>
<DIV class=3Dtable-caption><SPAN class=3Dtable-label>Table 1</SPAN>=20
<P id=3Dp-8 class=3Dfirst-child>Summary of Clinical, =
Immunohistochemical, and=20
Molecular Data for 194 Patients With Glioblastoma Multiforme*</P><BR=20
class=3Dcaption-clear></DIV></DIV>
<H3>FISH and Immunohistochemical Analysis on Tissue Microarrays</H3>
<P id=3Dp-11 class=3Dh-lead>Tissue microarrays (TMAs) were constructed =
using=20
formalin-fixed, paraffin-embedded archival tissue blocks as reported=20
elsewhere.<SUP><A id=3Dxref-ref-15-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-15">15</A=
></SUP>=20
We included 5 nontumoral control samples (tissue from 4 normal brains =
and 1=20
tonsil). H&amp;E-stained full sections from each donor block were used =
for=20
morphologic selection of the representative areas of each case. </P>
<P id=3Dp-12>FISH and immunohistochemical analyses were performed on 194 =
GBMs=20
arrayed on TMAs, although the number of tissue cores (samples) valid for =

assessment varied on each specific analysis. </P>
<P id=3Dp-13>FISH assays on TMAs were performed using <EM>EGFR</EM>,=20
<EM>MDM2</EM>, and <EM>CDK4</EM> gene-specific bacterial artificial =
chromosome=20
clones (RP11-339F13 for <EM>EGFR</EM>, RP11-611O2 and RP11-450G15 for=20
<EM>MDM2</EM>, and RP11-571M6 for <EM>CDK4</EM>). In addition, control =
clones=20
for each of the genes tested were used: p275=CE=B17a (centromere of =
chromosome 7) and=20
RP11-467M14/RP11-467M14 (12p13). BAC clones were selected from the =
EnsEMBL (<A=20
href=3D"http://www.ensembl.org/">http://www.ensembl.org/</A>) and UCSC =
(<A=20
href=3D"http://www.genome.ucsc.edu/">http://www.genome.ucsc.edu/</A>) =
databases.=20
The specificity and location of the probes were confirmed by FISH on =
normal=20
metaphases before hybridization on the TMA. Gene and control probes were =
labeled=20
in red and green, respectively, and hybridized as previously =
described.<SUP><A=20
id=3Dxref-ref-15-2 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-15">15</A=
></SUP></P>
<P id=3Dp-14>Fluorescence signals were scored by two of us (Y.R. and =
B.M.) in=20
accordance with the criteria described in previous reports.<SUP><A=20
id=3Dxref-ref-19-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-19">19</A=
>,<A=20
id=3Dxref-ref-20-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-20">20</A=
></SUP>=20
For each sample, only well-defined nuclei were analyzed, and the numbers =
of=20
single-copy gene and control probe signals were scored. Gene =
amplification was=20
considered positive when 5 or more unbalanced gene copies were detected =
in more=20
than 5% of tumor cells. </P>
<P id=3Dp-15>Immunohistochemical assays were performed on =
depar-affinized TMA=20
sections as reported elsewhere.<SUP><A id=3Dxref-ref-15-3 =
class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-15">15</A=
></SUP>=20
The primary antibodies used were EGFR (Oncogene Research Products, =
Boston, MA),=20
p53 (Novocastra Laboratories, Newcastle upon Tyne, England), CDK6 (BD=20
Biosciences Pharmingen, San Diego, CA), PTEN (DAKO Denmark A/S, =
Glostrup,=20
Denmark), and p21 (Oncogene Research Products). </P>
<DIV class=3Dh-p-wrap>
<H3><EM>p53</EM> Mutational Analysis </H3>
<P id=3Dp-16 class=3Dh-lead>Genomic DNA was extracted from =
paraffin-embedded=20
microdissected sections as previously described.<SUP><A =
id=3Dxref-ref-21-1=20
class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-21">21</A=
></SUP>=20
Mutations in exons 5 to 8 of the <EM>p53</EM> gene were screened by =
direct=20
sequencing in an ABI PRISM 310 DNA Analyzer (Applied Biosystems, Foster =
City,=20
CA) according to the manufacturer=E2=80=99s instructions. Primers and =
polymerase chain=20
reaction conditions were the same as reported previously.<SUP><A=20
id=3Dxref-ref-22-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-22">22</A=
></SUP></P></DIV>
<DIV class=3Dh-p-wrap>
<H3>Statistical Analysis</H3>
<P id=3Dp-17 class=3Dh-lead>Univariate and multivariate Cox regression =
analyses were=20
performed to identify the clinical and molecular variables related to =
survival.=20
Hazard ratios (HRs) and their associated 95% confidence intervals were =
computed=20
for selected variables using standard options for the Cox regression =
analysis.=20
Kaplan-Meier survival curves and Pearson correlation coefficients were =
also=20
calculated. All analyses were done using SPSS, version 11.0 (SPSS, =
Chicago, IL).=20
</P></DIV></DIV>
<DIV id=3Dsec-6 class=3D"section results">
<DIV class=3Dsection-nav><A class=3Dprev-section-link title=3D"Materials =
and Methods"=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#sec-1"><SPAN>=
Previous=20
Section</SPAN></A><A class=3Dnext-section-link title=3DDiscussion=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#sec-9"><SPAN>=
Next=20
Section</SPAN></A></DIV>
<H2>Results</H2>
<H3>Molecular Findings</H3>
<P id=3Dp-18 class=3Dh-lead><EM>EGFR</EM> gene amplification was =
detected in 75=20
(51.0%) of 147 GBMs, and strong EGFR overexpression was detected in 120 =
(65.9%)=20
of 182 GBMs. There was a significant correlation between the FISH and=20
immunophenotypic results for EGFR (=CF=81 =3D 0.526; <EM>P</EM> &lt; =
.01). Therefore,=20
taking into account any of these techniques, EGFR was altered in 127 =
(76.0%) of=20
167 GBMs. Strong and extensive nuclear immunostaining for p53 protein =
was found=20
in 21 (11.2%) of 187 GBMs <STRONG><A id=3Dxref-fig-1-1 class=3Dxref-fig=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#F1">Image=20
1</A></STRONG> (<A id=3Dxref-table-wrap-1-2 class=3Dxref-table=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#T1">Table =
1</A>).=20
Concurrent alteration of EGFR and p53 was found in 12 of 21 =
p53-immunopositive=20
samples. A subsequent mutational analysis confirmed <EM>p53</EM> =
mutation in 8=20
of the tumors available for analysis <A id=3Dxref-table-wrap-2-1 =
class=3Dxref-table=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#T2">Table =
2</A> (<A=20
id=3Dxref-fig-1-2 class=3Dxref-fig=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#F1">Image =
1</A>). </P>
<DIV id=3DT2 class=3Dtable>
<DIV class=3Dtable-inline>
<DIV class=3Dcallout><SPAN>View this table:</SPAN>
<UL class=3Dcallout-links>
  <LI><A=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/T2.expansion.html"=
>In=20
  this window</A>=20
  <LI><A class=3Din-nw=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/T2.expansion.html"=
>In a=20
  new window</A> </LI></UL></DIV></DIV>
<DIV class=3Dtable-caption><SPAN class=3Dtable-label>Table 2</SPAN>=20
<P id=3Dp-19 class=3Dfirst-child><EM>p53</EM> Mutations in Glioblastoma =
Multiforme=20
Samples With Concurrent EGFR-p53 Alteration </P><BR=20
class=3Dcaption-clear></DIV></DIV>
<DIV id=3DF1 class=3D"fig pos-float">
<DIV class=3Dfig-inline><A=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F1.expansion.html"=
><IMG=20
alt=3D"Image 1"=20
src=3D"http://ajcp.ascpjournals.org/content/131/2/257/F1.small.gif"></A>
<DIV class=3Dcallout><SPAN>View larger version:</SPAN>
<UL class=3Dcallout-links>
  <LI><A=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F1.expansion.html"=
>In=20
  this window</A>=20
  <LI><A class=3Din-nw=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F1.expansion.html"=
>In a=20
  new window</A> </LI></UL></DIV></DIV>
<DIV class=3Dfig-caption><SPAN class=3Dfig-label>Image 1</SPAN>=20
<P id=3Dp-21 class=3Dfirst-child><STRONG>A</STRONG>, Primary =
glioblastoma multiforme=20
(GBM) sample with strong positive expression of epidermal growth factor =
receptor=20
(EGFR; =C3=97400). <STRONG>B</STRONG>, The same primary GBM sample with =
positive=20
expression of p53 (=C3=97400). <STRONG>C</STRONG>, Mutational analysis =
of p53, exon=20
7. </P><BR class=3Dcaption-clear></DIV></DIV>
<P id=3Dp-22>To characterize the possible abnormalities of cell =
cycle=E2=80=93related=20
molecules involved in the EGFR and p53 pathways, we performed molecular =
analyses=20
of MDM2, CDK4, CDK6, p21, and PTEN in our series of primary GBM. We =
found=20
overexpression and/or amplification of <EM>MDM2</EM> and CDK4 in 20 =
(11.4%) of=20
175 tumors and 23 (19.8%) of 116 tumors, respectively. =
CDK6-immunopositive=20
expression was found in 56 (51.4%) of 109 primary GBMs, and there was =
absence of=20
expression of p21 and PTEN in 34 (37.0%) of 92 tumors and 68 (38.6%) of =
176=20
tumors, respectively (<A id=3Dxref-table-wrap-1-3 class=3Dxref-table=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#T1">Table =
1</A>). </P>
<H3>Survival Studies</H3>
<P id=3Dp-23 class=3Dh-lead>Univariate Cox regression analyses of =
clinical variables=20
in these series showed that age, P-KPS, extent of surgical resection,=20
radiotherapy, and chemotherapy were significantly associated with the =
clinical=20
outcome of the GBM patients <A id=3Dxref-table-wrap-3-1 =
class=3Dxref-table=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#T3">Table =
3</A>.=20
Subsequent univariate Cox regression analyses of the molecular variables =

revealed <EM>CDK4</EM> amplification to be a significant predictor of =
clinical=20
outcome, whereas none of the other molecular variables tested were =
independently=20
associated with patient survival (<A id=3Dxref-table-wrap-3-2 =
class=3Dxref-table=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#T3">Table =
3</A>). The=20
Kaplan-Meier survival curves indicated that <EM>CDK4</EM> amplification =
was=20
significantly associated with the worse overall survival of patients =
with GBM=20
(<EM>P</EM> =3D .01; log rank) <STRONG><A id=3Dxref-fig-2-1 =
class=3Dxref-fig=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#F2">Image=20
2</A></STRONG> and <A id=3Dxref-fig-3-1 class=3Dxref-fig=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#F3">Figure =
1</A>. </P>
<DIV id=3DT3 class=3Dtable>
<DIV class=3Dtable-inline>
<DIV class=3Dcallout><SPAN>View this table:</SPAN>
<UL class=3Dcallout-links>
  <LI><A=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/T3.expansion.html"=
>In=20
  this window</A>=20
  <LI><A class=3Din-nw=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/T3.expansion.html"=
>In a=20
  new window</A> </LI></UL></DIV></DIV>
<DIV class=3Dtable-caption><SPAN class=3Dtable-label>Table 3</SPAN>=20
<P id=3Dp-24 class=3Dfirst-child>Univariate Cox Regression Analyses in =
194 Primary=20
Glioblastoma Multiforme Cases</P><BR class=3Dcaption-clear></DIV></DIV>
<DIV id=3DF2 class=3D"fig pos-float">
<DIV class=3Dfig-inline><A=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F2.expansion.html"=
><IMG=20
alt=3D"Image 2"=20
src=3D"http://ajcp.ascpjournals.org/content/131/2/257/F2.small.gif"></A>
<DIV class=3Dcallout><SPAN>View larger version:</SPAN>
<UL class=3Dcallout-links>
  <LI><A=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F2.expansion.html"=
>In=20
  this window</A>=20
  <LI><A class=3Din-nw=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F2.expansion.html"=
>In a=20
  new window</A> </LI></UL></DIV></DIV>
<DIV class=3Dfig-caption><SPAN class=3Dfig-label>Image 2</SPAN>=20
<P id=3Dp-25 class=3Dfirst-child><EM>CDK4</EM> amplification observed in =
1 primary=20
glioblastoma multiforme. A gene-specific bacterial artificial chromosome =
(BAC)=20
clone containing <EM>CDK4</EM> (12q14) and a control BAC clone (12p13) =
were=20
labeled in red and green, respectively. </P><BR =
class=3Dcaption-clear></DIV></DIV>
<DIV id=3DF3 class=3D"fig pos-float">
<DIV class=3Dfig-inline><A=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F3.expansion.html"=
><IMG=20
alt=3D"Figure 1"=20
src=3D"http://ajcp.ascpjournals.org/content/131/2/257/F3.small.gif"></A>
<DIV class=3Dcallout><SPAN>View larger version:</SPAN>
<UL class=3Dcallout-links>
  <LI><A=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F3.expansion.html"=
>In=20
  this window</A>=20
  <LI><A class=3Din-nw=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F3.expansion.html"=
>In a=20
  new window</A> </LI></UL></DIV></DIV>
<DIV class=3Dfig-caption><SPAN class=3Dfig-label>Figure 1</SPAN>=20
<P id=3Dp-26 class=3Dfirst-child>Kaplan-Meier survival curve comparing =
positive and=20
negative gene amplification for <EM>CDK4</EM> with survival in patients =
with=20
glioblastoma multiforme. The median overall survival times were 5.5 and =
10=20
months for cases with positive and negative <EM>CDK4</EM>-amplified =
samples,=20
respectively. <EM>P</EM> =3D .01; log-rank. </P><BR=20
class=3Dcaption-clear></DIV></DIV>
<P id=3Dp-27>In addition, multivariate analysis involving the =
significant clinical=20
and molecular variables associated with survival probability revealed =
that age=20
(<EM>P</EM> &lt; .05; HR, 2.89; 95% confidence interval [CI], =
1.01=E2=80=938.23),=20
radiotherapy (<EM>P</EM> &lt; .05; HR, 6.93; 95% CI, =
1.59=E2=80=9330.20), and=20
<EM>CDK4</EM> amplification (<EM>P</EM> &lt; .05; HR, 3.15; 95% CI, =
1.04=E2=80=939.56)=20
continued to be independent prognostic factors. </P>
<P id=3Dp-28>To analyze the influence of concomitant p53 and EGFR =
alterations=20
studied by FISH and/or immunohistochemical analysis on outcome, we used=20
Kaplan-Meier survival curves. The analysis of the immunophenotypic =
pattern of=20
p53 in the EGFR=E2=80=93 and EGFR+ GBM subgroups revealed that p53 =
expression was=20
significantly associated with poorer survival in the EGFR+ subgroup =
(amplified=20
and/or overexpressed; <EM>P</EM> &lt; .01; log-rank test). Likewise, the =

analysis of EGFR in the p53-immunopositive and p53-immunonegative =
subgroups=20
revealed that EGFR alteration (amplified and/or overexpressed) was =
significantly=20
associated with a worse clinical outcome in the p53-immunopositive =
subgroup=20
(<EM>P</EM> &lt; .01; log-rank test) <A id=3Dxref-fig-4-1 =
class=3Dxref-fig=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#F4">Figure =
2</A>. </P>
<DIV id=3DF4 class=3D"fig pos-float">
<DIV class=3Dfig-inline><A=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F4.expansion.html"=
><IMG=20
alt=3D"Figure 2"=20
src=3D"http://ajcp.ascpjournals.org/content/131/2/257/F4.small.gif"></A>
<DIV class=3Dcallout><SPAN>View larger version:</SPAN>
<UL class=3Dcallout-links>
  <LI><A=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F4.expansion.html"=
>In=20
  this window</A>=20
  <LI><A class=3Din-nw=20
  =
href=3D"http://ajcp.ascpjournals.org/content/131/2/257/F4.expansion.html"=
>In a=20
  new window</A> </LI></UL></DIV></DIV>
<DIV class=3Dfig-caption><SPAN class=3Dfig-label>Figure 2</SPAN>=20
<P id=3Dp-29 class=3Dfirst-child>Survival analysis in glioblastoma =
multiforme.=20
<STRONG>A</STRONG>, Kaplan-Meier analysis comparing positive and =
negative=20
protein expression of p53 with survival in epidermal growth factor =
receptor=20
(EGFR)+ samples (amplified and/or overexpressed). <EM>P</EM> &lt; .01; =
log-rank.=20
<STRONG>B</STRONG>, Kaplan-Meier survival curve comparing EGFR status,=20
determined by using fluorescence in situ hybridization and/or=20
immunohistochemical analysis, with survival in p53-immunopositive =
samples.=20
<EM>P</EM> &lt; .01; log-rank. </P><BR =
class=3Dcaption-clear></DIV></DIV></DIV>
<DIV id=3Dsec-9 class=3D"section discussion">
<DIV class=3Dsection-nav><A class=3Dprev-section-link title=3DResults=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#sec-6"><SPAN>=
Previous=20
Section</SPAN></A><A class=3Dnext-section-link title=3DAcknowledgments=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ack-1"><SPAN>=
Next=20
Section</SPAN></A></DIV>
<H2>Discussion</H2>
<P id=3Dp-30>GBM can be classified as a primary or secondary tumor on =
the basis of=20
clinical features. This distinction is supported by the finding of =
mutually=20
exclusive EGFR and p53 alterations. However, coexistence of <EM>p53</EM> =

mutation and <EM>EGFR</EM> amplification in subsets of primary GBM =
tumors has=20
been described.<SUP><A id=3Dxref-ref-9-2 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-9">9</A>=E2=
=80=93<A=20
id=3Dxref-ref-11-2 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-11">11</A=
></SUP>=20
Moreover, Thomas and coworkers<SUP><A id=3Dxref-ref-23-1 =
class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-23">23</A=
></SUP>=20
reported that the glioma cell line SKMG-3 had an unusual genotype in =
which a=20
<EM>p53</EM> gene mutation coexisted with an amplified <EM>EGFR</EM> =
gene. In=20
the present study, we detected EGFR alteration in most of the tumors of =
this=20
series (76.0%), and p53 nuclear immunostaining was found in 11.2% of the =
tumors.=20
In addition, concurrent EGFR and p53 alterations were found in =
approximately=20
half of the p53-immunopositive tumors. Therefore, our results are in =
agreement=20
with the mutually exclusive pattern of EGFR and p53 in primary GBMs, but =
also=20
support the existence of a minority subgroup with concurrence of EGFR =
and p53=20
aberrations.<SUP><A id=3Dxref-ref-10-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-10">10</A=
>,<A=20
id=3Dxref-ref-11-3 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-11">11</A=
></SUP></P>
<P id=3Dp-31>The series of 194 primary GBMs analyzed in this study was =
tested and=20
validated by performing univariate Cox regression analysis using =
clinical data,=20
such as age, KPS, or extent of surgical resection, that were previously=20
associated with better outcome.<SUP><A id=3Dxref-ref-24-1 =
class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-24">24</A=
>=E2=80=93<A=20
id=3Dxref-ref-27-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-27">27</A=
></SUP>=20
Survival analysis of molecular data revealed <EM>CDK4</EM> amplification =
to be=20
associated with worse survival in GBM. It is important to note that =
multivariate=20
analysis including significant variables pointed out <EM>CDK4</EM> =
amplification=20
as an independent marker with prognostic value. An aberrant function of=20
<EM>CDK4</EM> would be involved in the abnormalities of the p16/RB/CDK4 =
pathway,=20
which have been previously correlated with astrocytic differentiation =
and the=20
poorer survival of patients with glioma.<SUP><A id=3Dxref-ref-28-1 =
class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-28">28</A=
>,<A=20
id=3Dxref-ref-29-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-29">29</A=
></SUP></P>
<P id=3Dp-32>The prognostic significance of EGFR and p53 alteration in=20
astrocytomas has always been a cause of controversy. Several reports =
have=20
concluded that EGFR amplification and/or overexpression predicts shorter =

survival,<SUP><A id=3Dxref-ref-30-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-30">30</A=
>=E2=80=93<A=20
id=3Dxref-ref-33-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-33">33</A=
></SUP>=20
whereas others have found no statistically significant =
relationship.<SUP><A=20
id=3Dxref-ref-34-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-34">34</A=
>,<A=20
id=3Dxref-ref-35-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-35">35</A=
></SUP>=20
As with EGFR, no consensus has been reached about the prognostic value =
of p53.=20
Some groups claim no association with survival,<SUP><A =
id=3Dxref-ref-32-1=20
class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-32">32</A=
>,<A=20
id=3Dxref-ref-34-2 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-34">34</A=
>,<A=20
id=3Dxref-ref-35-2 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-35">35</A=
></SUP>=20
whereas others propose that p53 alteration is a negative or positive =
prognostic=20
factor.<SUP><A id=3Dxref-ref-30-2 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-30">30</A=
>,<A=20
id=3Dxref-ref-36-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-36">36</A=
></SUP>=E2=80=93<SUP><A=20
id=3Dxref-ref-38-1 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-38">38</A=
></SUP>=20
In our study, EGFR and p53 were not factors independently associated =
with=20
survival. However, we detected for the first time that the GBM subgroup =
showing=20
simultaneous alteration of EGFR and p53 pathways was characterized by a =
worse=20
clinical outcome. Thus, our findings may support the hypothesis that =
mutant=20
<EM>p53</EM> induces genomic instability in this fraction of tumors and, =

consequently, <EM>EGFR</EM> gene amplification.<SUP><A =
id=3Dxref-ref-10-2=20
class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-10">10</A=
></SUP>=20
The simultaneous deregulation of the EGFR and p53 pathways may indicate =
a=20
relevant cell cycle deregulation that leads to more aggressive GBM =
formation.=20
</P>
<P id=3Dp-33>In contrast with our results, Simmons and coworkers<SUP><A=20
id=3Dxref-ref-33-2 class=3Dxref-bibr=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-33">33</A=
></SUP>=20
reported that younger patients with GBM (&lt;55 years, median series =
age), with=20
EGFR overexpression and p53 immunonegativity had a statistically =
significant=20
worse survival. This contradictory result may be due to the higher =
percentage of=20
p53 immunopositivity in the younger patients (57% vs 16%) or in the =
overall=20
series (49% vs 11.2%), when both data sets are compared. In this sense, =
our=20
series showed high EGFR alteration (76.0%) and low p53 expression =
(11.2%), which=20
are suggestive of a primary GBM pattern. </P>
<P id=3Dp-34><EM>CDK4</EM> amplification is associated with a =
significantly worse=20
survival in patients with GBM, and it is a molecular variable showing=20
independent prognostic value. Likewise, although EGFR and p53 are =
mutually=20
exclusive, there is a primary GBM subgroup exhibiting concurrent EGFR =
and p53=20
alterations that is associated with a poorer clinical outcome. These =
findings=20
may help in the search for useful prognostic and therapeutic markers. =
However,=20
additional studies will almost certainly highlight further the genetic=20
complexity and importance of the EGFR-p53 positive subgroup in the =
biology of=20
primary GBMs. </P></DIV>
<DIV id=3Dack-1 class=3D"section ack">
<DIV class=3Dsection-nav><A class=3Dprev-section-link title=3DDiscussion =

href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#sec-9"><SPAN>=
Previous=20
Section</SPAN></A><A class=3Dnext-section-link title=3DFootnotes=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#fn-group-1"><=
SPAN>Next=20
Section</SPAN></A></DIV>
<H2>Acknowledgments</H2>
<P id=3Dp-35>We acknowledge the assistance of Lydia Sanchez (Spanish =
National=20
Research Center, Madrid) and Juan Fernando Garc=C3=ADa (M.D. Anderson =
International,=20
Madrid) with the immunohistochemical studies and of Yolanda Vicente =
(supported=20
by Fondo de Investigaciones Sanitarias, Madrid, FIS CA06/0006) and Elena =
G=C3=B3mez=20
(Virgen de la Salud Hospital, Toledo) for their technical assistance. =
</P></DIV>
<DIV id=3Dfn-group-1 class=3D"section fn-group">
<DIV class=3Dsection-nav><A class=3Dprev-section-link =
title=3DAcknowledgments=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ack-1"><SPAN>=
Previous=20
Section</SPAN></A><A class=3Dnext-section-link title=3DReferences=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#ref-list-1"><=
SPAN>Next=20
Section</SPAN></A></DIV>
<H2>Footnotes</H2>
<UL>
  <LI id=3Dfn-4 class=3Dfn-supported-by>
  <P id=3Dp-36>Supported in part by grants FIS-02/3006, PI070662, =
CA07/00119, and=20
  INT 07/028 from the Fondo de Investigaciones Sanitarias, Madrid; and =
JI05012,=20
  FISCAM PI-2006/29, and SESCAM 04032 from the Consejer=C3=ADa de =
Sanidad Junta de=20
  Comunidades de Castilla-La Mancha, Toledo. </P></LI>
  <LI id=3Dcopyright-statement-1 =
class=3Dcopyright-statement>Copyright=C2=A9 by the=20
  American Society for Clinical Pathology </LI></UL></DIV>
<DIV id=3Dref-list-1 class=3D"section ref-list">
<DIV class=3Dsection-nav><A class=3Dprev-section-link title=3DFootnotes=20
href=3D"http://ajcp.ascpjournals.org/content/131/2/257.full#fn-group-1"><=
SPAN>Previous=20
Section</SPAN></A>
<DIV class=3Dnav-placeholder></DIV></DIV>
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Content-Location: http://ajcp.ascpjournals.org/content/131/2/hw-global-dynamic-elements.css

DIV#hovering-abs-close {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
DIV#hovering-abs-close A {
	COLOR: #444; FONT-WEIGHT: bold; TEXT-DECORATION: none
}
DIV#hovering-abs-close A:hover {
	COLOR: black; FONT-WEIGHT: bold; TEXT-DECORATION: none
}
DIV#hovering-abs-close A SPAN {
	FONT-SIZE: 0.66em
}
DIV#hovering-abs H1 {
	PADDING-BOTTOM: 0.4em; MARGIN: 0.5em; PADDING-LEFT: 0.4em; =
PADDING-RIGHT: 0.4em; FONT-SIZE: 1em; PADDING-TOP: 0.4em
}
DIV#hovering-abs {
	Z-INDEX: 99; BORDER-BOTTOM: #333 1px solid; BORDER-LEFT: #333 1px =
solid; PADDING-BOTTOM: 0.4em; LINE-HEIGHT: 1.3; BACKGROUND-COLOR: =
#d8d8d8; MARGIN: 0.5em; PADDING-LEFT: 0.4em; WIDTH: 405px; =
PADDING-RIGHT: 0.4em; FONT-SIZE: 0.66em; OVERFLOW: auto; BORDER-TOP: =
#333 1px solid; BORDER-RIGHT: #333 1px solid; PADDING-TOP: 0.4em; LEFT: =
140px
}
DIV#hovering-abs DIV {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
DIV#hovering-abs P {
	PADDING-BOTTOM: 0.4em; MARGIN: 0px; PADDING-LEFT: 1em; PADDING-RIGHT: =
1em; PADDING-TOP: 0.4em
}
DIV#hovering-abs-ptr {
	BACKGROUND-IMAGE: url(../../shared/img/misc/abs-ptr.gif); Z-INDEX: 99; =
POSITION: absolute; PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; =
WIDTH: 32px; PADDING-RIGHT: 0px; BACKGROUND-POSITION: right bottom; =
HEIGHT: 32px; TOP: 1200px; PADDING-TOP: 0px; LEFT: 510px
}
DIV#hovering-abs OL {
	DISPLAY: none
}
DIV#hovering-abs UL {
	DISPLAY: none
}
DIV#hovering-abs DIV.fn-group {
	DISPLAY: none
}
DIV.hw-gen-page .view-more {
	BORDER-BOTTOM: #dcdcdc 1px solid; TEXT-ALIGN: center; BORDER-LEFT: =
#dcdcdc 1px solid; PADDING-BOTTOM: 0.4em; BACKGROUND-COLOR: white; =
WIDTH: 1.3em; DISPLAY: block; FLOAT: left; HEIGHT: 0.9em; MARGIN-LEFT: =
-2px; FONT-SIZE: 0.83em; BORDER-TOP: #dcdcdc 1px solid; CURSOR: pointer; =
FONT-WEIGHT: bold; MARGIN-RIGHT: 5px; BORDER-RIGHT: #dcdcdc 1px solid; =
TEXT-DECORATION: none
}
DIV.hw-gen-page .view-more:hover {
	BACKGROUND-COLOR: #383838; COLOR: white
}

------=_NextPart_000_0000_01C98BC5.4FC41B00
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: 7bit
Content-Location: http://ajcp.ascpjournals.org/shared/css/hw-global.css

@import url( hw-global-sidebars.css );
@import url( hw-global-elements.css );
@import url( hw-global-citation.css );
@import url( hw-global-classes.css );
@import url( hw-global-dynamic-elements.css );

------=_NextPart_000_0000_01C98BC5.4FC41B00
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://ajcp.ascpjournals.org/content/131/2/ui.css

BODY {
	BACKGROUND-COLOR: #cccccc; MARGIN: 0px auto; WIDTH: 1000px; FONT-SIZE: =
100%
}
DIV.hw-gen-page {
	POSITION: relative; PADDING-BOTTOM: 0px; BACKGROUND-COLOR: white; =
MARGIN: 10px auto 0px; PADDING-LEFT: 0px; WIDTH: 960px; PADDING-RIGHT: =
0px; FONT-FAMILY: "Lucida Sans Unicode", Arial, "Lucida Grande", Tahoma, =
Verdana, Helvetica, sans-serif; COLOR: #403838; PADDING-TOP: 0px
}
DIV.hw-gen-page A {
	COLOR: #202088; TEXT-DECORATION: none
}
DIV.hw-gen-page A:link {
	COLOR: #202088; TEXT-DECORATION: none
}
DIV.hw-gen-page A:visited {
	COLOR: #581858; TEXT-DECORATION: none
}
DIV.hw-gen-page A:hover {
	BORDER-BOTTOM: #202088 1px dotted; COLOR: #202088; TEXT-DECORATION: =
none
}
DIV.hw-gen-page A:active {
	COLOR: #202088; TEXT-DECORATION: none
}
#header A {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; COLOR: #404040; BORDER-LEFT-STYLE: none
}
#header A:link {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; COLOR: #404040; BORDER-LEFT-STYLE: none
}
#footer A {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; COLOR: #404040; BORDER-LEFT-STYLE: none
}
#footer A:link {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; COLOR: #404040; BORDER-LEFT-STYLE: none
}
#header A:visited {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; COLOR: #404040; BORDER-LEFT-STYLE: none
}
#footer A:visited {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; COLOR: #404040; BORDER-LEFT-STYLE: none
}
#header UL.button-list A:hover {
	BORDER-BOTTOM: #404040 1px dotted; COLOR: #404040; TEXT-DECORATION: =
none
}
#header DIV.header-ac-elements A:hover {
	BORDER-BOTTOM: #404040 1px dotted; COLOR: #404040; TEXT-DECORATION: =
none
}
#header DIV.header-qs A:hover {
	BORDER-BOTTOM: #404040 1px dotted; COLOR: #404040; TEXT-DECORATION: =
none
}
#footer A:hover {
	BORDER-BOTTOM: #404040 1px dotted; COLOR: #404040; TEXT-DECORATION: =
none
}
#header A:active {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; COLOR: #404040; BORDER-LEFT-STYLE: none
}
#footer A:active {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; COLOR: #404040; BORDER-LEFT-STYLE: none
}
#col-2 A {
	COLOR: #404040
}
#col-2 A:link {
	COLOR: #404040
}
#col-3 A {
	COLOR: #404040
}
#col-3 A:link {
	COLOR: #404040
}
#col-2 A:visited {
	COLOR: #581858
}
#col-3 A:visited {
	COLOR: #581858
}
#col-2 A:hover {
	BORDER-BOTTOM: #404040 1px dotted; COLOR: #404040; TEXT-DECORATION: =
none
}
#col-3 A:hover {
	BORDER-BOTTOM: #404040 1px dotted; COLOR: #404040; TEXT-DECORATION: =
none
}
#col-2 A:active {
	COLOR: #404040
}
#col-3 A:active {
	COLOR: #404040
}
#col-3 UL.cover-announce A {
	COLOR: #2f5085
}
#col-3 UL.cover-announce A:link {
	COLOR: #2f5085
}
#col-3 UL.cover-announce A:visited {
	COLOR: #2f5085
}
#col-3 UL.cover-announce A:hover {
	BORDER-BOTTOM: #2f5085 1px dotted; COLOR: #2f5085; TEXT-DECORATION: =
none
}
#col-3 UL.cover-announce A:active {
	COLOR: #2f5085
}
DIV.sidebar A:hover {
	TEXT-DECORATION: none
}
DIV#header {
	BACKGROUND-IMAGE: url(/shared/img/standard-design/design2/corner.gif); =
BORDER-BOTTOM-STYLE: none; MARGIN-TOP: 0px; WIDTH: 960px; =
BACKGROUND-REPEAT: no-repeat; BACKGROUND-POSITION: right top
}
#header #main-logo {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; MARGIN: 15px 0px =
0px 24px; BORDER-TOP-STYLE: none; BORDER-LEFT-STYLE: none
}
#header UL.home-page-ads {
	POSITION: absolute; PADDING-BOTTOM: 5px; LIST-STYLE-TYPE: none; MARGIN: =
1.5em 0px 35px; PADDING-LEFT: 10px; WIDTH: 250px; PADDING-RIGHT: 10px; =
TOP: 0px; PADDING-TOP: 5px; LEFT: 450px
}
#header UL.home-page-ads LI {
	TEXT-ALIGN: center; PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: =
0px; WIDTH: 250px; PADDING-RIGHT: 0px; HEIGHT: 40px; PADDING-TOP: 0px
}
#header UL.home-page-ads LI IMG {
	BORDER-BOTTOM: #aaaaaa 1px solid; BORDER-LEFT: #aaaaaa 1px solid; =
BACKGROUND-COLOR: #e0e0e0; WIDTH: 250px; DISPLAY: block; HEIGHT: 40px; =
BORDER-TOP: #aaaaaa 1px solid; BORDER-RIGHT: #aaaaaa 1px solid
}
#header .banner-ads {
	POSITION: absolute; PADDING-BOTTOM: 5px; LIST-STYLE-TYPE: none; MARGIN: =
1.5em 0px 35px; PADDING-LEFT: 10px; WIDTH: 250px; PADDING-RIGHT: 10px; =
TOP: 0px; PADDING-TOP: 5px; LEFT: 450px
}
.banner-ads#header UL {
	LIST-STYLE-TYPE: none
}
#header .banner-ads LI {
	TEXT-ALIGN: center; PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: =
0px; PADDING-RIGHT: 0px; PADDING-TOP: 0px
}
#header .banner-ads LI IMG {
	BORDER-BOTTOM: #aaaaaa 1px solid; BORDER-LEFT: #aaaaaa 1px solid; =
BACKGROUND-COLOR: #e0e0e0; DISPLAY: block; BORDER-TOP: #aaaaaa 1px =
solid; BORDER-RIGHT: #aaaaaa 1px solid
}
#header .header-buttons {
	PADDING-BOTTOM: 0px; LIST-STYLE-TYPE: none; MARGIN: 5px 0px 8px 24px; =
PADDING-LEFT: 0px; PADDING-RIGHT: 0px; PADDING-TOP: 0px
}
#header .header-buttons {
	COLOR: #0083a8
}
#header .header-buttons A {
	COLOR: #0083a8
}
#header .header-buttons LI {
	BORDER-LEFT: #666666 1px solid; PADDING-BOTTOM: 0px; MARGIN: 0px; =
PADDING-LEFT: 0.4em; PADDING-RIGHT: 0.4em; DISPLAY: inline; PADDING-TOP: =
0px
}
#header .header-buttons LI SPAN {
	FONT-SIZE: 87.5%; FONT-WEIGHT: bold
}
#header .header-buttons LI.first {
	PADDING-LEFT: 0px; BORDER-LEFT-STYLE: none
}
#header .corner {
	POSITION: absolute; TOP: 0px; RIGHT: 0px
}
#header .inst-branding {
	BORDER-LEFT: #c8c8c8 1px dashed
}
#header #hdr-login {
	BORDER-LEFT: #c8c8c8 1px dashed
}
#header .header-qs {
	BORDER-LEFT: #c8c8c8 1px dashed
}
#header .inst-branding {
	POSITION: absolute; PADDING-LEFT: 5px; HEIGHT: 30px; TOP: 0px; LEFT: =
756px
}
#header #authstring {
	POSITION: absolute; TEXT-ALIGN: center; WIDTH: 756px; DISPLAY: block; =
PADDING-TOP: 5px; LEFT: 0%
}
#header #authstring {
	COLOR: white
}
#header #authstring LI {
	BORDER-BOTTOM-STYLE: none; PADDING-BOTTOM: 2px; BORDER-RIGHT-STYLE: =
none; LIST-STYLE-TYPE: none; PADDING-LEFT: 2px; PADDING-RIGHT: 2px; =
BORDER-TOP-STYLE: none; FONT-SIZE: 87.5%; BORDER-LEFT-STYLE: none; =
PADDING-TOP: 2px
}
#header #authstring A {
	COLOR: white
}
#header #hdr-login {
	POSITION: absolute; PADDING-BOTTOM: 5px; PADDING-LEFT: 10px; =
PADDING-RIGHT: 0px; TOP: 30px; PADDING-TOP: 0px; LEFT: 756px
}
#header #hdr-login FORM LABEL {
	WHITE-SPACE: nowrap; FONT-SIZE: 87.5%
}
#header #hdr-login INPUT {
	WHITE-SPACE: nowrap; FONT-SIZE: 87.5%
}
#header #hdr-login {
	COLOR: #0083a8
}
#header #hdr-login LABEL {
	PADDING-BOTTOM: 2px; MARGIN: 0px; PADDING-LEFT: 0px; WIDTH: 80px; =
PADDING-RIGHT: 0px; DISPLAY: block; FONT-SIZE: 87.5%; PADDING-TOP: 5px
}
#header #hdr-login INPUT {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; WIDTH: 110px; PADDING-RIGHT: =
0px; HEIGHT: 15px; FONT-SIZE: 87.5%; PADDING-TOP: 0px
}
#header .header-qs INPUT {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; WIDTH: 110px; PADDING-RIGHT: =
0px; HEIGHT: 15px; FONT-SIZE: 87.5%; PADDING-TOP: 0px
}
#header #hdr-login INPUT#hdr-login-signin {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 5px; WIDTH: auto; PADDING-RIGHT: =
0px; HEIGHT: auto; VERTICAL-ALIGN: bottom; PADDING-TOP: 0px
}
#header .bar {
	PADDING-LEFT: 20px; WIDTH: 980px; HEIGHT: 60px; MARGIN-LEFT: -20px
}
#footer .bar {
	PADDING-LEFT: 20px; WIDTH: 980px; HEIGHT: 60px; MARGIN-LEFT: -20px
}
#header .bar {
	BACKGROUND-COLOR: #0083a8
}
#footer .bar {
	BACKGROUND-COLOR: #0083a8
}
#header .bar-inner {
	WIDTH: 960px; HEIGHT: 60px; COLOR: white
}
#header .bar-inner {
	BACKGROUND-COLOR: #00a0cc
}
#footer .footer-group {
	BACKGROUND-COLOR: #00a0cc
}
#header .header-qs {
	Z-INDEX: 1; POSITION: absolute; PADDING-BOTTOM: 7px; MARGIN: 0px; =
PADDING-LEFT: 10px; WIDTH: 195px; PADDING-RIGHT: 5px; DISPLAY: block; =
HEIGHT: 40px; TOP: 112px; PADDING-TOP: 10px; LEFT: 756px
}
#header .header-qs LABEL {
	POSITION: absolute; LEFT: -9999px
}
#header #hdr-login-signin-label {
	POSITION: absolute; LEFT: -9999px
}
#header #header-qs-search-label {
	POSITION: absolute; LEFT: -9999px
}
#header .header-qs A {
	LINE-HEIGHT: 1.5; WHITE-SPACE: nowrap; COLOR: white; FONT-SIZE: 87.5%
}
#header DIV.adv-search-link A:hover {
	BORDER-BOTTOM: white 1px dotted; COLOR: white
}
#header DIV.adv-search-link A:visited {
	COLOR: white
}
#header DIV.adv-search-link A:active {
	COLOR: white
}
#header DIV.adv-search-link A:link {
	COLOR: white
}
#header .header-qs INPUT#header-qs-search-go {
	WIDTH: 27px; HEIGHT: auto; MARGIN-LEFT: 0.5em; VERTICAL-ALIGN: bottom
}
#content-block {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 5px; WIDTH: 518px; PADDING-RIGHT: =
20px; FLOAT: left; PADDING-TOP: 0px
}
#col-2 {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
#col-3 {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
#col-2 {
	MARGIN: 0px; WIDTH: 200px; PADDING-RIGHT: 13px; FLOAT: left; OVERFLOW: =
hidden; BORDER-RIGHT: #aaaaaa 1px dashed
}
#content-block {
	BACKGROUND-COLOR: white; COLOR: #403838
}
.pagetype-proxied #content-block {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; WIDTH: 756px; PADDING-RIGHT: =
0px; BORDER-RIGHT: #aaaaaa 1px dashed; PADDING-TOP: 0px
}
.proxied-column-display#proxied-contents #col-main {
	WIDTH: 518px; PADDING-RIGHT: 10px; FLOAT: left; OVERFLOW: hidden
}
.proxied-column-display#proxied-contents #col-2 {
	FLOAT: left; BORDER-RIGHT: 0px solid
}
#proxied-contents {
	PADDING-LEFT: 15px; PADDING-RIGHT: 10px
}
.proxied-column-display#proxied-contents {
	PADDING-RIGHT: 0px
}
.proxied-column-display#proxied-contents .search-nav A {
	COLOR: black; FONT-WEIGHT: normal
}
.pagetype-proxied #header .banner-ads UL {
	MARGIN: 1em 5%
}
.pagetype-proxied #col-3 .content-box H3 {
	PADDING-BOTTOM: 0.3em; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; FONT-SIZE: 1.2em; PADDING-TOP: 0.3em
}
#proxied-contents TH {
	FONT-WEIGHT: bold
}
#proxied-contents TABLE {
	LINE-HEIGHT: 110%
}
#proxied-contents TD {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
TR {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
TH {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
P {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
OL {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
UL {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
#proxied-contents H1 {
	TEXT-ALIGN: left; LINE-HEIGHT: 120%
}
H3 {
	TEXT-ALIGN: left; LINE-HEIGHT: 120%
}
DL {
	TEXT-ALIGN: left; LINE-HEIGHT: 120%
}
DT {
	TEXT-ALIGN: left; LINE-HEIGHT: 120%
}
DD {
	TEXT-ALIGN: left; LINE-HEIGHT: 120%
}
#proxied-contents .proxied-collections H1 {
	MARGIN-TOP: 0.5em
}
#proxied-contents H1 {
	TEXT-ALIGN: left; LINE-HEIGHT: 100%
}
H3 {
	TEXT-ALIGN: left; LINE-HEIGHT: 100%
}
DL {
	TEXT-ALIGN: left; LINE-HEIGHT: 100%
}
DT {
	TEXT-ALIGN: left; LINE-HEIGHT: 100%
}
DD {
	TEXT-ALIGN: left; LINE-HEIGHT: 100%
}
#proxied-contents #col-3 P {
	MARGIN: 1em 5%
}
PRE {
	MARGIN: 1em 5%
}
LABEL {
	MARGIN: 1em 5%
}
UL {
	MARGIN: 1em 5%
}
OL {
	MARGIN: 1em 5%
}
DL {
	MARGIN: 1em 5%
}
FIELDSET {
	MARGIN: 1em 5%
}
ADDRESS {
	MARGIN: 1em 5%
}
#col-2 #cb-art-nav LI {
	MARGIN-TOP: 0.4em
}
#col-2 DIV.sidebar-issue-nav {
	MARGIN: 5px 0px 0px 5px
}
#col-2 DIV.article-nav {
	MARGIN: 5px 0px 0px 5px
}
#col-2 DIV.search-nav {
	MARGIN: 5px 0px 0px 5px
}
#col-3 {
	TEXT-ALIGN: left; PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; =
WIDTH: 203px; PADDING-RIGHT: 0px; FLOAT: left; OVERFLOW: visible; =
PADDING-TOP: 0px
}
#col-3 {
	BACKGROUND-COLOR: white
}
#col-3 * {
	POSITION: static
}
#col-3 DIV {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
DIV.content-box A:link {
	COLOR: #404040; TEXT-DECORATION: none
}
DIV.content-box A:visited {
	COLOR: #404040; TEXT-DECORATION: none
}
DIV.content-box A:hover {
	COLOR: #404040; TEXT-DECORATION: none
}
DIV.content-box A:active {
	COLOR: #404040; TEXT-DECORATION: none
}
#col-3 UL.pub-links LI A:link {
	BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none
}
#col-3 UL.pub-links LI A:visited {
	BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none
}
#col-3 UL.pub-links LI A:hover {
	BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none
}
#col-3 UL.pub-links LI A:active {
	BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none
}
#col-3 UL.ex-pub-links LI A:link {
	BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none
}
#col-3 UL.ex-pub-links LI A:visited {
	BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none
}
#col-3 UL.ex-pub-links LI A:hover {
	BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none
}
#col-3 UL.ex-pub-links LI A:active {
	BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none
}
#col-3 UL.pub-links {
	POSITION: relative; PADDING-BOTTOM: 0px; MARGIN: 10px 0px; =
PADDING-LEFT: 0px; PADDING-RIGHT: 0px; OVERFLOW: hidden; PADDING-TOP: =
0px; LEFT: -7px
}
#col-3 UL.pub-links {
	LIST-STYLE-TYPE: none
}
#col-3 UL.pub-links {
	COLOR: #0083a8
}
#col-3 UL.pub-links LI A {
	COLOR: #0083a8
}
#col-3 UL.pub-links LI {
	BORDER-BOTTOM: #aaaaaa 1px; POSITION: static; BORDER-LEFT: #aaaaaa 1px =
dashed; PADDING-BOTTOM: 10px; BACKGROUND-COLOR: white; MARGIN: 0px; =
PADDING-LEFT: 4px; PADDING-RIGHT: 5px; FONT-SIZE: 0.85em; BORDER-TOP: =
#aaaaaa 1px dashed; FONT-WEIGHT: bold; BORDER-RIGHT: #aaaaaa 1px dashed; =
PADDING-TOP: 10px
}
#col-3 UL.pub-links LI A {
	BACKGROUND-IMAGE: =
url(/shared/img/standard-design/design2/jnl-link-bullet.gif); =
LINE-HEIGHT: 1.2; PADDING-LEFT: 1em; DISPLAY: block; BACKGROUND-REPEAT: =
no-repeat; BACKGROUND-POSITION: left top
}
#col-3 UL.pub-links LI.last {
	BORDER-BOTTOM: #aaaaaa 1px dashed
}
#col-3 .sidebar-icon-group {
	PADDING-BOTTOM: 20px; PADDING-LEFT: 20px; PADDING-RIGHT: 20px; =
PADDING-TOP: 20px
}
#col-3 .sidebar-icon-group A:hover {
	BORDER-BOTTOM: 0px solid
}
#col-3 UL.ex-pub-links {
	POSITION: relative; PADDING-BOTTOM: 0px; MARGIN: 10px 0px; =
PADDING-LEFT: 0px; PADDING-RIGHT: 0px; OVERFLOW: hidden; PADDING-TOP: =
0px; LEFT: -7px
}
#col-3 UL.ex-pub-links {
	LIST-STYLE-TYPE: none
}
#col-3 UL.ex-pub-links {
	COLOR: #0083a8
}
#col-3 UL.ex-pub-links LI A {
	COLOR: #0083a8
}
#col-3 UL.ex-pub-links LI {
	BORDER-BOTTOM: #aaaaaa 1px; POSITION: static; BORDER-LEFT: #aaaaaa 1px =
dashed; PADDING-BOTTOM: 10px; BACKGROUND-COLOR: white; MARGIN: 0px; =
PADDING-LEFT: 4px; PADDING-RIGHT: 5px; FONT-SIZE: 75%; BORDER-TOP: =
#aaaaaa 1px dashed; FONT-WEIGHT: bold; BORDER-RIGHT: #aaaaaa 1px dashed; =
PADDING-TOP: 10px
}
#col-3 UL.ex-pub-links LI A {
	BACKGROUND-IMAGE: =
url(/shared/img/standard-design/design2/jnl-link-bullet.gif); =
LINE-HEIGHT: 1.2; PADDING-LEFT: 1em; DISPLAY: block; BACKGROUND-REPEAT: =
no-repeat; BACKGROUND-POSITION: left top
}
#col-3 UL.ex-pub-links LI.last {
	BORDER-BOTTOM: #aaaaaa 1px dashed
}
#col-3 UL.tower-ads {
	PADDING-BOTTOM: 0em; LIST-STYLE-TYPE: none; MARGIN: 0em; PADDING-LEFT: =
0em; PADDING-RIGHT: 0em; LIST-STYLE-IMAGE: none; PADDING-TOP: 0em
}
#col-3 UL.tower-ads LI IMG {
	PADDING-BOTTOM: 0px; MARGIN: 0em auto 0.5em; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; DISPLAY: block; PADDING-TOP: 0px
}
#col-3 .publisher-icon {
=09
}
#footer {
	TEXT-ALIGN: left; WIDTH: 960px
}
#footer {
	BACKGROUND-IMAGE: url(/shared/img/standard-design/design2/dash.gif); =
PADDING-BOTTOM: 20px; BACKGROUND-COLOR: white; BACKGROUND-REPEAT: =
repeat-y; BACKGROUND-POSITION: 756px 0px; CLEAR: both
}
#footer UL {
	LIST-STYLE-TYPE: none; FONT-SIZE: 80%
}
#footer .bar {
	POSITION: static; CLEAR: both
}
#footer .footer-group {
	HEIGHT: 60px; COLOR: white; FONT-WEIGHT: bold
}
#footer .footer-group P {
	PADDING-BOTTOM: 10px; PADDING-LEFT: 10px; PADDING-RIGHT: 10px; =
PADDING-TOP: 10px
}
#footer .footer-col-left {
	WIDTH: 756px; FLOAT: left; BORDER-RIGHT: #c8c8c8 1px dashed
}
#footer .footer-col-right {
	WIDTH: 203px; FLOAT: left
}
#footer .footer-col-right UL {
	PADDING-BOTTOM: 0px; LIST-STYLE-TYPE: none; MARGIN: 0px; PADDING-LEFT: =
0px; PADDING-RIGHT: 0px; DISPLAY: block; PADDING-TOP: 8px
}
#footer .footer-col-right LI {
	BORDER-BOTTOM-STYLE: none; TEXT-ALIGN: left; PADDING-BOTTOM: 4px; =
BORDER-RIGHT-STYLE: none; PADDING-LEFT: 10px; PADDING-RIGHT: 10px; =
DISPLAY: block; BORDER-TOP-STYLE: none; BORDER-LEFT-STYLE: none; =
PADDING-TOP: 4px
}
#footer .copyright {
	MARGIN: 1em auto; FONT-SIZE: 80%
}
#footer A:hover {
	BORDER-BOTTOM: white 1px dotted; COLOR: white
}
#skip-link {
	POSITION: absolute; LEFT: -9999px
}
#header #hdr-login {
	HEIGHT: 75px
}
DIV.hw-gen-page .view-more {
	BORDER-BOTTOM: gray 1px solid; BORDER-LEFT: gray 1px solid; BORDER-TOP: =
gray 1px solid; BORDER-RIGHT: gray 1px solid
}
#header #hdr-login LABEL#hdr-login-signin-label {
	DISPLAY: inline
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://ajcp.ascpjournals.org/content/131/2/home-page.css

#pageid-home #content-block DIV.main-block {
	WIDTH: 350px; MARGIN-BOTTOM: 10px; FLOAT: left
}
#pageid-home #content-block .main-block LI {
	MARGIN: 0px
}
#pageid-home #content-block .main-block UL {
	MARGIN: 0px
}
#pageid-home #content-block H2 {
	FONT-FAMILY: Arial, "Lucida Grande", "Lucida Sans Unicode", Tahoma, =
Verdana, Helvetica, sans-serif; FONT-SIZE: 1.2em
}
#pageid-home #content-block H3 {
	FONT-FAMILY: Arial, "Lucida Grande", "Lucida Sans Unicode", Tahoma, =
Verdana, Helvetica, sans-serif; FONT-SIZE: 1.2em
}
#pageid-home #content-block H2 {
	COLOR: #0083a8
}
#pageid-home #content-block H3 {
	COLOR: #0083a8
}
#pageid-home #content-block H2 A {
	COLOR: #0083a8
}
#pageid-home #content-block DIV.home-announce A {
	COLOR: #0083a8
}
#content-block DIV.home-announce A:hover {
	BORDER-RIGHT-WIDTH: 0px; BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: =
0px; BORDER-LEFT-WIDTH: 0px; TEXT-DECORATION: underline
}
#pageid-home #content-block H2.current-issue {
	PADDING-BOTTOM: 20px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 20px
}
#pageid-home #content-block A.current-issue-link {
	BORDER-BOTTOM: #0083a8 1px dotted
}
#pageid-home #content-block .cover-img-wrap {
	BACKGROUND-IMAGE: =
url(/shared/img/standard-design/design2/cover_shadow.png); POSITION: =
relative; BACKGROUND-REPEAT: no-repeat; TOP: -25px; LEFT: 20px
}
#pageid-home #cover-image {
	POSITION: relative; LEFT: 5px
}
#pageid-home #cover-image IMG {
	BORDER-BOTTOM-STYLE: none; PADDING-BOTTOM: 5px; BORDER-RIGHT-STYLE: =
none; BACKGROUND-COLOR: white; PADDING-LEFT: 5px; PADDING-RIGHT: 5px; =
BORDER-TOP-STYLE: none; BORDER-LEFT-STYLE: none; PADDING-TOP: 5px
}
#pageid-home DIV.cover-img-wrap A:hover {
	BORDER-BOTTOM: 0px solid
}
#pageid-home #content-block DIV.home-announce-wrapper {
	BACKGROUND-IMAGE: =
url(/shared/img/standard-design/design2/announce_shadow_top.2.png); =
POSITION: relative; MARGIN: 0px; WIDTH: 282px; BACKGROUND-REPEAT: =
no-repeat; LEFT: 20px
}
#pageid-home #content-block DIV.home-announce-wrapper-bottomTRYWITHOUT {
	BACKGROUND-IMAGE: =
url(/shared/img/standard-design/design2/announce_shadow_bottom.png); =
WIDTH: 282px; BACKGROUND-REPEAT: no-repeat; BACKGROUND-POSITION: left =
bottom; HEIGHT: 10px
}
#pageid-home #content-block DIV.home-announce-bottom {
	POSITION: relative; PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; WIDTH: =
282px; PADDING-RIGHT: 0px; MARGIN-BOTTOM: 20px; BACKGROUND: =
url(/shared/img/standard-design/design2/announce_shadow_bottom.3.png) =
#e0e0e0 no-repeat left bottom; HEIGHT: 10px; PADDING-TOP: 0px; LEFT: =
20px
}
#pageid-home #content-block DIV.home-announce {
	BORDER-BOTTOM: white 0px solid; POSITION: relative; BORDER-LEFT: white =
5px solid; BACKGROUND-COLOR: #e0e0e0; PADDING-LEFT: 10px; WIDTH: 242px; =
PADDING-RIGHT: 10px; BORDER-TOP: white 0px solid; BORDER-RIGHT: white =
5px solid; PADDING-TOP: 10px; LEFT: 5px
}
#pageid-home #content-block DIV.home-announce-spaceTRYWITHOUT {
	BORDER-BOTTOM: white 0px solid; POSITION: relative; BORDER-LEFT: white =
5px solid; PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #e0e0e0; PADDING-LEFT: =
0px; WIDTH: 262px; PADDING-RIGHT: 0px; BORDER-TOP: white 0px solid; =
BORDER-RIGHT: white 5px solid; PADDING-TOP: 0px; LEFT: 5px
}
#pageid-home #content-block DIV.home-announce {
	FONT-WEIGHT: bold
}
#pageid-home #content-block UL {
	LIST-STYLE-TYPE: none
}
#pageid-home #content-block UL.jnl-links {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
MARGIN-LEFT: 0px; PADDING-TOP: 0px
}
#pageid-home #content-block UL.jnl-links P {
	MARGIN: 0px 0px 20px; FONT-SIZE: 1.2em
}
#pageid-home #content-block UL.jnl-links LI.empty H3 {
	MARGIN: 0px 0px 20px
}
#pageid-home #content-block .in-this-issue H2 {
	BORDER-TOP: #0083a8 1px dotted; PADDING-TOP: 10px
}
#pageid-home #content-block .in-this-issue UL {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
#pageid-home #content-block .in-this-issue UL LI {
	PADDING-BOTTOM: 5px; LINE-HEIGHT: 1.3; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; PADDING-TOP: 0px
}
#pageid-home #content-block .in-this-issue UL A {
	BORDER-LEFT: #0083a8 5px solid; PADDING-LEFT: 0.5em
}
#pageid-home #content-block .content-block-clear {
	WIDTH: 100%; HEIGHT: 2px; CLEAR: both
}
#pageid-home #content-block {
	PADDING-BOTTOM: 0px; BACKGROUND-COLOR: white; MARGIN: 0px; =
PADDING-LEFT: 0px; WIDTH: 756px; PADDING-RIGHT: 0px; FONT-FAMILY: =
arial,sans-serif; FLOAT: left; COLOR: #666666; OVERFLOW: visible; =
BORDER-RIGHT: #aaaaaa 1px dashed; PADDING-TOP: 0px
}
#pageid-home #col-3 {
	TEXT-ALIGN: left; PADDING-BOTTOM: 0px; BACKGROUND-COLOR: white; MARGIN: =
0px; PADDING-LEFT: 0px; WIDTH: 203px; PADDING-RIGHT: 0px; FLOAT: left; =
OVERFLOW: visible; PADDING-TOP: 0px
}
#pageid-home #intl-content {
	TEXT-ALIGN: center; FONT-SIZE: 1em
}
#pageid-home #intl-content A {
	COLOR: #333333
}
#pageid-home #intl-content SPAN.intl-help {
	FONT-SIZE: 0.75em
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: 7bit
Content-Location: http://ajcp.ascpjournals.org/shared/css/standard-designs/design2/main.css

@import url( ui.css );
@import url( home-page.css );

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://ajcp.ascpjournals.org/content/131/2/hw-ascpjnls-sidebars.css

#col-2 DIV#docked-slugline {
	FONT-WEIGHT: normal
}
#col-2 DIV#slugline {
	FONT-WEIGHT: normal
}
#col-2 {
	BACKGROUND-COLOR: #eeeeee
}
#col-2 SPAN.slug-vol {
	FONT-STYLE: italic
}
#col-2 ABBR.slug-jnl-abbrev {
	FONT-STYLE: italic
}
DIV#docked-slugline .slug-pub-date {
	FONT-WEIGHT: normal
}
DIV#docked-slugline .slug-pages {
	FONT-WEIGHT: normal
}
DIV#slugline CITE .slug-pub-date {
	FONT-WEIGHT: normal
}
DIV#slugline CITE .slug-pages {
	FONT-WEIGHT: normal
}
#col-2 DIV.sidebar {
	BORDER-BOTTOM: #553a37 1px solid; PADDING-BOTTOM: 5px; =
BORDER-RIGHT-WIDTH: 0px; MARGIN: 10px 7px 5px; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; BORDER-TOP-WIDTH: 0px; FONT-SIZE: 90%; =
BORDER-LEFT-WIDTH: 0px; PADDING-TOP: 5px
}
#col-2 .issue-sections {
	BORDER-TOP: #553a37 0px solid
}
#col-2 SPAN.open-access-note {
	COLOR: #f88a00
}
#col-2 DIV.sidebar H4 {
	PADDING-BOTTOM: 0.3em; MARGIN: 0pt; PADDING-LEFT: 0pt; PADDING-RIGHT: =
0pt; FONT-SIZE: 1em; FONT-WEIGHT: bold; PADDING-TOP: 0.3em
}
#col-2 DIV.sidebar P {
	LINE-HEIGHT: 1.2; MARGIN: 0.5em 5%
}
#col-2 DIV.sidebar UL {
	LINE-HEIGHT: 1.2; MARGIN: 0.5em 5%
}
#col-2 DIV.sidebar OL {
	LINE-HEIGHT: 1.2; MARGIN: 0.5em 5%
}
#col-2 DIV.sidebar-qs {
	BORDER-BOTTOM: #553a37 1px solid; BORDER-LEFT: #553a37 1px solid; =
PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #cccccc; MARGIN: 0px 0px 5px 7px; =
PADDING-LEFT: 5px; PADDING-RIGHT: 5px; FONT-SIZE: 88%; BORDER-TOP: =
#553a37 1px solid; BORDER-RIGHT: #553a37 1px solid; PADDING-TOP: 5px
}
#col-2 P.qs-instructions {
	MARGIN: 3px 0px
}
#col-2 DIV.sidebar-qs FORM.searchbox {
	MARGIN: 0px
}
#col-2 DIV.sidebar-qs DIV {
	MARGIN: 0px
}
#col-2 DIV.sidebar-qs IMG {
	PADDING-BOTTOM: 0px; MARGIN: 5px 0px 0px 5px; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; PADDING-TOP: 0px
}
#col-2 FORM.searchbox #issue-search-go {
	BORDER-BOTTOM-STYLE: solid; BORDER-RIGHT-STYLE: solid; =
BACKGROUND-COLOR: #608e3a; WIDTH: 60px; BORDER-TOP-STYLE: solid; COLOR: =
#ffffff; MARGIN-LEFT: 5px; FONT-SIZE: 0.8em; BORDER-LEFT-STYLE: solid; =
FONT-WEIGHT: bold
}
#col-2 DIV.content-box DIV.cb-section SPAN.access {
	COLOR: #cc0000
}
#col-2 DIV.content-box {
	BACKGROUND-COLOR: #96ad83
}
DIV.sidebar A:active {
	COLOR: #608e3a; TEXT-DECORATION: none
}
DIV.sidebar A:link {
	COLOR: #608e3a; TEXT-DECORATION: none
}
DIV.sidebar A:visited {
	COLOR: #608e3a; TEXT-DECORATION: none
}
DIV#col-3 A {
	COLOR: #608e3a; TEXT-DECORATION: none
}
DIV#col-3 #sidebar-section A.hover {
	BORDER-BOTTOM: #608e3a 1px dotted
}
DIV#col-3 #sidebar-global-nav A.hover {
	BORDER-BOTTOM: #608e3a 1px dotted
}
DIV.sidebar A:hover {
	TEXT-DECORATION: none
}
#col-3 #sidebar-current-issue {
	BORDER-BOTTOM: #96ad83 2px solid; BORDER-LEFT: #96ad83 2px solid; =
PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #eaeaea; MARGIN: 5px 10px; =
PADDING-LEFT: 5px; PADDING-RIGHT: 5px; BORDER-TOP: #96ad83 2px solid; =
BORDER-RIGHT: #96ad83 2px solid; PADDING-TOP: 5px
}
#col-3 #sidebar-section {
	BORDER-BOTTOM: #96ad83 4px solid; BORDER-LEFT: #96ad83 4px solid; =
PADDING-BOTTOM: 5px; LINE-HEIGHT: 1.5em; BACKGROUND-COLOR: #eaeaea; =
LIST-STYLE-TYPE: none; MARGIN: 5px 10px; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; FONT-SIZE: 83.33%; BORDER-TOP: #96ad83 4px solid; =
BORDER-RIGHT: #96ad83 4px solid; PADDING-TOP: 5px
}
#col-3 #sidebar-current-issue DIV.cb-contents {
	PADDING-BOTTOM: 0px
}
#col-3 DIV.content-box .cb-contents LI {
	TEXT-ALIGN: left
}
#col-3 DIV.content-box DIV.current-issue {
	TEXT-ALIGN: center; PADDING-BOTTOM: 5px; MARGIN: 0px; PADDING-LEFT: =
0px; PADDING-RIGHT: 0px; PADDING-TOP: 5px
}
#col-3 DIV.content-box DIV.current-issue IMG {
	BORDER-BOTTOM: #333300 1px solid; BORDER-LEFT: #333300 1px solid; =
BORDER-TOP: #333300 1px solid; BORDER-RIGHT: #333300 1px solid
}
#col-3 DIV.content-box DIV.current-issue A:hover {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; BORDER-LEFT-STYLE: none
}
#col-3 DIV#sidebar-global-nav {
	POSITION: relative; PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: =
0px; PADDING-RIGHT: 0px; PADDING-TOP: 0px
}
#col-3 UL.pub-links {
	COLOR: #666666
}
#col-3 UL.pub-links LI A {
	COLOR: #666666
}
#col-3 .sidebar-icon-group IMG {
	DISPLAY: block; MARGIN-LEFT: auto; MARGIN-RIGHT: auto; PADDING-TOP: =
10px
}
#col-3 UL.ex-pub-links LI {
	BACKGROUND-COLOR: #cddfee
}
#col-3 UL.ex-pub-links LI A {
	BACKGROUND-IMAGE: url(/local/img/ccjm-jnl-link-bullet.gif); COLOR: =
#4f5862
}
#col-3 DIV.cb-section {
	BORDER-BOTTOM: #553a37 1px solid; PADDING-BOTTOM: 3px; MARGIN: 0pt; =
PADDING-LEFT: 0pt; PADDING-RIGHT: 10px; PADDING-TOP: 3px
}
.article-nav {
	BORDER-BOTTOM: #5f907b 1px dotted; BORDER-LEFT: #5f907b 1px dotted; =
BACKGROUND-COLOR: #d6e0c7; BORDER-TOP: #5f907b 1px dotted; BORDER-RIGHT: =
#5f907b 1px dotted
}
.search-nav {
	BORDER-BOTTOM: #5f907b 1px dotted; BORDER-LEFT: #5f907b 1px dotted; =
BACKGROUND-COLOR: #d6e0c7; BORDER-TOP: #5f907b 1px dotted; BORDER-RIGHT: =
#5f907b 1px dotted
}
.article-nav A:hover {
	COLOR: #333333; TEXT-DECORATION: none
}
.search-nav A:hover {
	COLOR: #333333; TEXT-DECORATION: none
}
#col-2 A.prev-results-link {
	BACKGROUND-COLOR: #eeec83
}
#col-2 A.next-results-link {
	BACKGROUND-COLOR: #eeec83
}
DIV.content-box DIV.cb-section .notice A {
	BACKGROUND-COLOR: #eeec83
}
DIV.content-box DIV.cb-section A.pdf-direct-link {
	BACKGROUND-COLOR: #eeec83
}
FORM.searchbox #sidebar-search-input {
	WIDTH: 90px; MARGIN-LEFT: 0px
}
FORM.searchbox #issue-search-input {
	WIDTH: 90px; MARGIN-LEFT: 0px
}
#proxied-contents #col-2 #col-3 H1 {
	TEXT-ALIGN: left; LINE-HEIGHT: 100%
}
H3 {
	TEXT-ALIGN: left; LINE-HEIGHT: 100%
}
DL {
	TEXT-ALIGN: left; LINE-HEIGHT: 100%
}
DT {
	TEXT-ALIGN: left; LINE-HEIGHT: 100%
}
DD {
	TEXT-ALIGN: left; LINE-HEIGHT: 100%
}
#proxied-contents #col-2 #col-3p {
	MARGIN: 1em 5%
}
PRE {
	MARGIN: 1em 5%
}
LABEL {
	MARGIN: 1em 5%
}
UL {
	MARGIN: 1em 5%
}
OL {
	MARGIN: 1em 5%
}
DL {
	MARGIN: 1em 5%
}
FIELDSET {
	MARGIN: 1em 5%
}
ADDRESS {
	MARGIN: 1em 5%
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://ajcp.ascpjournals.org/publisher/css/hw-ascpjnls-global.css

@import url( hw-ascpjnls-sidebars.css );
DIV.hw-gen-page {
	COLOR: black
}
#content-block {
	COLOR: black
}
#header .header-buttons {
	COLOR: #000000
}
#header .header-buttons A {
	COLOR: #000000
}
#header #authstring A {
	COLOR: black
}
#header #authstring {
	COLOR: black
}
#header #authstring LI {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 5px; PADDING-RIGHT: 5px; =
PADDING-TOP: 0px
}
#header .header-qs A {
	COLOR: black
}
#header DIV.adv-search-link A:hover {
	BORDER-BOTTOM: black 1px dotted; COLOR: black
}
#footer .footer-group {
	COLOR: black
}
#header .header-buttons {
	COLOR: #000000
}
#header .header-buttons A {
	COLOR: #000000
}
#header #hdr-login {
	COLOR: #000000
}
#header .header-buttons LI SPAN {
	FONT-SIZE: 80%
}
#header #hdr-login {
	COLOR: #27258d
}
#footer .footer-group {
	COLOR: #666666
}
#footer .footer-group A {
	COLOR: #666666
}
#header FORM.searchbox DIV.adv-search-link A {
	COLOR: #666666
}
#col-3 .sidebar-icon-group {
	PADDING-BOTTOM: 20px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
PADDING-TOP: 20px
}
#col-3 UL.pub-links {
	COLOR: #666666
}
#col-3 UL.pub-links LI A {
	COLOR: #666666
}
FORM#adv-search H1 {
	COLOR: #608e3a
}
FORM#adv-search .search-date-range {
	COLOR: black
}
#content-block LEGEND {
	PADDING-BOTTOM: 0.1em; BACKGROUND-COLOR: #591b5e; MARGIN: 0pt; =
PADDING-LEFT: 0.4em; PADDING-RIGHT: 0.4em; FONT-SIZE: 0.95em; =
PADDING-TOP: 0.1em
}
#content-block DIV.article {
	COLOR: #000000
}
DIV.hw-gen-page {
	COLOR: black
}
#content-block {
	COLOR: black
}
#header .bar {
	BACKGROUND-COLOR: #dedede
}
#footer .bar {
	BACKGROUND-COLOR: #dedede
}
#header .bar-inner {
	BACKGROUND-COLOR: #ededed
}
#footer .footer-group {
	BACKGROUND-COLOR: #ededed
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://ajcp.ascpjournals.org/content/131/2/hw-ajcp-sidebars.css

.article-nav {
	BORDER-BOTTOM: #5f907b 1px dotted; BORDER-LEFT: #5f907b 1px dotted; =
BACKGROUND-COLOR: #f5aec2; BORDER-TOP: #5f907b 1px dotted; BORDER-RIGHT: =
#5f907b 1px dotted
}
.search-nav {
	BORDER-BOTTOM: #5f907b 1px dotted; BORDER-LEFT: #5f907b 1px dotted; =
BACKGROUND-COLOR: #f5aec2; BORDER-TOP: #5f907b 1px dotted; BORDER-RIGHT: =
#5f907b 1px dotted
}
#col-2 DIV.content-box {
	BACKGROUND-COLOR: #ac0031
}
#col-3 #sidebar-current-issue {
	BORDER-BOTTOM: #ac0031 2px solid; BORDER-LEFT: #ac0031 2px solid; =
BACKGROUND-COLOR: #eaeaea; BORDER-TOP: #ac0031 2px solid; BORDER-RIGHT: =
#ac0031 2px solid
}
#col-2 FORM.searchbox #issue-search-go {
	BACKGROUND-COLOR: #ac0031
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://ajcp.ascpjournals.org/local/css/hw-ajcp-global.css

@import url( hw-ajcp-sidebars.css );
#proxied-contents UL.supplement-list {
	LIST-STYLE-TYPE: none; MARGIN: 1em
}
#proxied-contents UL.supplement-list LI {
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------=_NextPart_000_0000_01C98BC5.4FC41B00
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		modClass($this, 'collapsible', 'collapsed');=0A=
	}=0A=
	else if ($this.hasClass('collapsible')) {=0A=
		modClass($this, 'collapsed', 'collapsible');=0A=
	}=0A=
	var $toToggle =3D $this.find(hideTagExpr);=0A=
	if ($toToggle.length) {=0A=
		$toToggle.toggle();=0A=
	}=0A=
	var $rewrite =3D $this.find(modTextTagExpr);=0A=
	if ($rewrite.length) {=0A=
		$rewrite.each(=0A=
			function(i, e) {=0A=
				var txt =3D $(e).text();=0A=
				if (txt =3D=3D '+') {=0A=
					$(e).empty().append('-');=0A=
				}=0A=
				else if (txt =3D=3D '-') {=0A=
					$(e).empty().append('+');=0A=
				}=0A=
			}=0A=
		);=0A=
	}=0A=
}=0A=
=0A=
=0A=
function updateFormInput(labelMatchString, inputMatchString, =
defaultColorString, textColorString) {=0A=
	if ((defaultColorString =3D=3D null) || (defaultColorString =3D=3D '')) =
{=0A=
		defaultColorString =3D "#A0A0A0";=0A=
	}=0A=
	if ((textColorString =3D=3D null) || (textColorString =3D=3D '')) {=0A=
		textColorString =3D "black";=0A=
	}=0A=
	var label =3D $(labelMatchString);=0A=
	var input =3D $(inputMatchString);=0A=
	if (input.length) {=0A=
		if ((label.length) && (input.val() =3D=3D '')) {=0A=
			input.val(label.text()).css("color",defaultColorString);=0A=
		}=0A=
		input.focus(=0A=
			function(e) {=0A=
				if ((label.length) &&  ($(this).val() =3D=3D label.text())) {=0A=
					$(this).val('').css("color",textColorString);=0A=
				}=0A=
			}=0A=
		);=0A=
		input.blur(=0A=
			function(e) {=0A=
				if ((label.length) && ($(this).val() =3D=3D '')) {=0A=
					$(this).val(label.text()).css("color",defaultColorString);=0A=
				}=0A=
			}=0A=
		);=0A=
	}=0A=
	var parentForm =3D label.parents("form").eq(0);=0A=
	if (parentForm.length) {=0A=
		parentForm.submit(=0A=
			function() {=0A=
				if ((label.length) && (input.length) && (input.val() =3D=3D =
label.text())) {=0A=
					input.val('');=0A=
				}=0A=
				return true;=0A=
			}=0A=
		);=0A=
	}=0A=
}=0A=
=0A=
function checkUnloadedImgs(lookupRule) {=0A=
	if (lookupRule =3D=3D null) {=0A=
		lookupRule =3D "img";=0A=
	}=0A=
	var imageNum =3D -1;=0A=
	var images =3D $(lookupRule);=0A=
	if (images.length) {=0A=
		imageNum =3D 0;=0A=
		images.each(=0A=
			function() {=0A=
				//if ($(this).height() =3D=3D 0) {=0A=
				if (this.offsetHeight =3D=3D 0) {=0A=
					imageNum++;=0A=
				}=0A=
			}=0A=
		);=0A=
	}=0A=
	return imageNum;=0A=
}=0A=
=0A=
function fixHeightForImages(iter, numImagesToLoad, lookupRule) {=0A=
	if (lookupRule =3D=3D null) {=0A=
		lookupRule =3D "img";=0A=
	}=0A=
	if (numImagesToLoad > 0) {=0A=
		var lastNumImagesToLoad =3D numImagesToLoad;=0A=
		var newNumImagesToLoad =3D checkUnloadedImgs(lookupRule);=0A=
		if (newNumImagesToLoad < lastNumImagesToLoad) {=0A=
			fixColHeights(1);=0A=
			if ((newNumImagesToLoad > 0) && (iter < 10)) {=0A=
				setTimeout("fixHeightForImages(" + (iter+1) + "," + =
newNumImagesToLoad + ",'" + lookupRule + "')", 1000);=0A=
			}=0A=
		}=0A=
	}=0A=
}=0A=
function getColInfo(colJQEl) {=0A=
	var colInfo =3D { valid: false, height: 0, bottom: 0, lastElBottom: 0, =
extra: 0, fix: false };=0A=
	if (colJQEl.length) {=0A=
		var c =3D colJQEl.children();=0A=
		if (c.length) {=0A=
			colInfo.height =3D getObjHeight(colJQEl);=0A=
			colInfo.bottom =3D colJQEl.offset().top + colInfo.height;=0A=
			var last =3D c.eq(c.length - 1);=0A=
			colInfo.lastElBottom =3D last.offset().top + getObjHeight(last);=0A=
			colInfo.extra =3D colInfo.bottom - colInfo.lastElBottom;=0A=
			colInfo.valid =3D true;=0A=
		}=0A=
	}=0A=
	return colInfo;=0A=
}=0A=
function checkColHeights() {=0A=
	var colToFix =3D 0;=0A=
	if (gIsMultiCol && ((gColOrigHeights.main + gColOrigHeights.col2 + =
gColOrigHeights.col3) > 0)) {=0A=
		var colMainInfo =3D getColInfo(gColMain);=0A=
		var col2Info =3D getColInfo(gCol2);=0A=
		var col3Info =3D getColInfo(gCol3);=0A=
		if (colMainInfo.valid && (gColLows.main =3D=3D 0)) { gColLows.main =3D =
colMainInfo.lastElBottom; }=0A=
		if (col2Info.valid && (gColLows.col2 =3D=3D 0)) { gColLows.col2 =3D =
col2Info.lastElBottom; }=0A=
		if (col3Info.valid && (gColLows.col3 =3D=3D 0)) { gColLows.col3 =3D =
col3Info.lastElBottom; }=0A=
		// check if low point in any column has changed=0A=
		if ((colMainInfo.valid) && (colMainInfo.lastElBottom !=3D =
gColLows.main)) {=0A=
			colMainInfo.fix =3D true;=0A=
		}=0A=
		if ((col2Info.valid) && (col2Info.lastElBottom !=3D gColLows.col2)) {=0A=
			col2Info.fix =3D true;=0A=
		}=0A=
		if ((col3Info.valid) && (col3Info.lastElBottom !=3D gColLows.col3)) {=0A=
			col3Info.fix =3D true;=0A=
		}=0A=
		// check for columns not having same bottom points=0A=
		if ((colMainInfo.valid) &&=0A=
			  (((col2Info.valid) && (colMainInfo.height < col2Info.height)) ||=0A=
			   ((col3Info.valid) && (colMainInfo.height < col3Info.height)))) {=0A=
			colMainInfo.fix =3D true;=0A=
		}=0A=
		if ((col2Info.valid) &&=0A=
			  (((colMainInfo.valid) && (col2Info.height < colMainInfo.height)) ||=0A=
			   ((col3Info.valid) && (col2Info.height < col3Info.height)))) {=0A=
			col2Info.fix =3D true;=0A=
		}=0A=
		if ((col3Info.valid) &&=0A=
			  (((colMainInfo.valid) && (col3Info.height < colMainInfo.height)) ||=0A=
			   ((col2Info.valid) && (col3Info.height < col2Info.height)))) {=0A=
			col3Info.fix =3D true;=0A=
		}=0A=
		=0A=
		if (colToFix > -1) {=0A=
			if (colMainInfo.valid && colMainInfo.fix) {=0A=
				if ((col2Info.valid && col2Info.fix) || (col3Info.valid && =
col3Info.fix)) {=0A=
					colToFix =3D -1;=0A=
				}=0A=
				else { colToFix =3D 1; }=0A=
			}=0A=
			else if (col2Info.valid && col2Info.fix) {=0A=
				if (col3Info.valid && col3Info.fix) {=0A=
					colToFix =3D -1;=0A=
				}=0A=
				else { colToFix =3D 2; }=0A=
			}=0A=
			else if (col3Info.valid && col3Info.fix) {=0A=
				colToFix =3D 3;=0A=
			}=0A=
		}=0A=
		=0A=
		if (colToFix !=3D 0) {=0A=
			fixColHeights(colToFix);=0A=
		}=0A=
		if (colMainInfo.valid) { gColLows.main =3D colMainInfo.lastElBottom; }=0A=
		if (col2Info.valid) { gColLows.col2 =3D col2Info.lastElBottom; }=0A=
		if (col3Info.valid) { gColLows.col3 =3D col3Info.lastElBottom; }=0A=
	}=0A=
	setTimeout("checkColHeights()", 5000);=0A=
}=0A=
function fixColHeights(colChanged) {=0A=
	if (gIsMultiCol) {=0A=
		var setAll =3D false;=0A=
		if (colChanged =3D=3D 1) {=0A=
			if (gColMain.length) {=0A=
				setObjHeight(gColMain,'auto');=0A=
				gColOrigHeights.main =3D getObjHeight(gColMain);=0A=
			}=0A=
		}=0A=
		else if (colChanged =3D=3D 2) {=0A=
			if (gCol2.length) {=0A=
				setObjHeight(gCol2,'auto');=0A=
				gColOrigHeights.col2 =3D getObjHeight(gCol2);=0A=
			}=0A=
		}=0A=
		else if (colChanged =3D=3D 3) {=0A=
			if (gCol3.length) {=0A=
				setObjHeight(gCol3,'auto');=0A=
				gColOrigHeights.col3 =3D getObjHeight(gCol3);=0A=
			}=0A=
		}=0A=
		else {=0A=
			if (colChanged =3D=3D -1) {=0A=
				// force resize all 3=0A=
				if (gColMain.length) { setObjHeight(gColMain,'auto'); }=0A=
				if (gCol2.length) { setObjHeight(gCol2,'auto'); }=0A=
				if (gCol3.length) { setObjHeight(gCol3,'auto'); }=0A=
			}=0A=
			gColOrigHeights.main =3D ((gColMain.length) ? =
(getObjHeight(gColMain)) : 0);=0A=
			gColOrigHeights.col2 =3D ((gCol2.length) ? (getObjHeight(gCol2)) : 0);=0A=
			gColOrigHeights.col3 =3D ((gCol3.length) ? (getObjHeight(gCol3)) : 0);=0A=
			setAll =3D true;=0A=
		}=0A=
		if ((gColOrigHeights.main + gColOrigHeights.col2 + =
gColOrigHeights.col3) > 0) {=0A=
			var colInfo;=0A=
			var maxH =3D Math.max(Math.max(gColOrigHeights.main, =
gColOrigHeights.col2), gColOrigHeights.col3);=0A=
			if ((gColMain.length) && ((gColOrigHeights.main < maxH) || =
(getObjHeight(gColMain) > maxH) || setAll)) {=0A=
				setObjHeight(gColMain, maxH);=0A=
				colInfo =3D getColInfo(gColMain);=0A=
				if (colInfo.valid) { gColLows.main =3D colInfo.lastElBottom; }=0A=
			}=0A=
			if ((gCol2.length) && ((gColOrigHeights.col2 < maxH) || =
(getObjHeight(gCol2) > maxH) || setAll)) {=0A=
				setObjHeight(gCol2, maxH);=0A=
				colInfo =3D getColInfo(gCol2);=0A=
				if (colInfo.valid) { gColLows.col2 =3D colInfo.lastElBottom; }=0A=
			}=0A=
			if ((gCol3.length) && ((gColOrigHeights.col3 < maxH) || =
(getObjHeight(gCol3) > maxH) || setAll)) {=0A=
				setObjHeight(gCol3, maxH);=0A=
				colInfo =3D getColInfo(gCol3);=0A=
				if (colInfo.valid) { gColLows.col3 =3D colInfo.lastElBottom; }=0A=
			}=0A=
		}=0A=
	}=0A=
}=0A=
=0A=
function getObjHeight(obj) {=0A=
	if (obj instanceof jQuery) {=0A=
		return obj.get(0).offsetHeight;=0A=
	}=0A=
	else if (obj instanceof Array) {=0A=
		return obj[0].offsetHeight;=0A=
	}=0A=
	else {=0A=
		return obj.offsetHeight;=0A=
	}=0A=
}=0A=
function getObjWidth(obj) {=0A=
	if (obj instanceof jQuery) {=0A=
		return obj.get(0).offsetWidth;=0A=
	}=0A=
	else if (obj instanceof Array) {=0A=
		return obj[0].offsetWidth;=0A=
	}=0A=
	else {=0A=
		return obj.offsetWidth;=0A=
	}=0A=
}=0A=
function setObjHeight(obj, h) {=0A=
	if (typeof(h) =3D=3D 'number') {=0A=
		h =3D "" + h + "px";=0A=
	}=0A=
	if (obj instanceof jQuery) {=0A=
		obj.get(0).style.height =3D h;=0A=
	}=0A=
	else if (obj instanceof Array) {=0A=
		obj[0].style.height =3D h;=0A=
	}=0A=
	else {=0A=
		obj.style.height =3D h;=0A=
	}=0A=
}=0A=
function setObjWidth(obj, w) {=0A=
	if (typeof(w) =3D=3D 'number') {=0A=
		w =3D "" + w + "px";=0A=
	}=0A=
	if (obj instanceof jQuery) {=0A=
		obj.get(0).style.width =3D w;=0A=
	}=0A=
	else if (obj instanceof Array) {=0A=
		obj[0].style.width =3D w;=0A=
	}=0A=
	else {=0A=
		obj.style.width =3D w;=0A=
	}=0A=
}=0A=
=0A=
/* num pixels top of page has been scrolled offscreen */=0A=
function getPageOffset() {=0A=
	var offset =3D new Object();=0A=
  if( typeof( window.pageXOffset ) =3D=3D 'number' ) {=0A=
    offset.x =3D window.pageXOffset;=0A=
    offset.y =3D window.pageYOffset;=0A=
  } else if( document.body && ( document.body.scrollTop ) ) {=0A=
    offset.x =3D document.body.scrollLeft;=0A=
    offset.y =3D document.body.scrollTop;=0A=
  } else if( document.documentElement && ( =
document.documentElement.scrollTop ) ) {=0A=
    offset.x =3D document.documentElement.scrollLeft;=0A=
    offset.y =3D document.documentElement.scrollTop;=0A=
  }=0A=
  return offset;=0A=
}=0A=
function getViewportDim() {=0A=
	var dim =3D new Object();=0A=
	// non-IE=0A=
	if (typeof window.innerWidth !=3D 'undefined') {=0A=
		dim.x =3D window.innerWidth;=0A=
		dim.y =3D window.innerHeight;=0A=
	}=0A=
	// IE6 in standards compliant mode=0A=
	else if (typeof document.documentElement !=3D 'undefined'=0A=
		&& typeof document.documentElement.clientWidth !=3D=0A=
		'undefined' && document.documentElement.clientWidth !=3D 0) {=0A=
		dim.x =3D document.documentElement.clientWidth;=0A=
		dim.y =3D document.documentElement.clientHeight;=0A=
	}=0A=
	// older versions of IE=0A=
	else {=0A=
		dim.x =3D document.getElementsByTagName('body')[0].clientWidth;=0A=
		dim.y =3D document.getElementsByTagName('body')[0].clientHeight;=0A=
	}=0A=
	return dim;=0A=
}=0A=
=0A=
function newWindowTargets() {=0A=
	var newWins =3D $("a.in-nw");=0A=
	if (newWins.length) {=0A=
		newWins.each(=0A=
			function() {=0A=
				var $this =3D $(this);=0A=
				$this.attr("target", "_blank");=0A=
				modClass($this, 'in-nw-vis', 'in-nw');=0A=
			}=0A=
		);=0A=
	}=0A=
}=0A=
=0A=
function addPartHeaders(req) {=0A=
	req.setRequestHeader('Accept', 'application/xhtml+xml');=0A=
	req.setRequestHeader('Range', 'part=3Dvariant-contents');=0A=
}=0A=
function addCommonHeaders(req) {=0A=
	if (typeof(callbackToken) !=3D 'undefined') {=0A=
		req.setRequestHeader('X-Token', callbackToken);=0A=
	}=0A=
}=0A=
function allowsCookies() {=0A=
	if (hasTestCookie()) {=0A=
		return true;=0A=
	}=0A=
	else {=0A=
		setTestCookie();=0A=
		return hasTestCookie();=0A=
	}=0A=
}=0A=
function setTestCookie() {=0A=
	setCookie('cks','allowed',7,'/',null,null);=0A=
}=0A=
function hasTestCookie() {=0A=
	var ckVal =3D getCookie('cks');=0A=
	if ((ckVal !=3D null) && (ckVal =3D=3D 'allowed')) {=0A=
		return true;=0A=
	}=0A=
	else {=0A=
		return false;=0A=
	}=0A=
}=0A=
=0A=
var UIPrefsCk=3D{=0A=
	name: "UIPrefs",=0A=
	expDays: 3652,=0A=
	path: '/'=0A=
};=0A=
=0A=
function prefDefined(name) {=0A=
	var val =3D getPrefValue(name);=0A=
	if (val =3D=3D null) {=0A=
		return false;=0A=
	}=0A=
	else {=0A=
		return true;=0A=
	}=0A=
}=0A=
function getPrefValue(name) {=0A=
	var prefArray =3D getPrefArray();=0A=
	if (prefArray) {=0A=
		name =3D convertPrefString(name);=0A=
		var match =3D name + ":";=0A=
		for (var i =3D 0; i < prefArray.length; i++) {=0A=
			if (prefArray[i].indexOf(match) =3D=3D 0) {=0A=
				return prefArray[i].substring(match.length);=0A=
			}=0A=
			else if (prefArray[i] =3D=3D name) {=0A=
				// name only, no value, return boolean true=0A=
				return true;=0A=
			}=0A=
		}=0A=
	}=0A=
	return null;=0A=
}=0A=
function convertPrefString(str) {=0A=
	if (str) {=0A=
		return str.replace(/[,:=3D]/g,"");=0A=
	}=0A=
	return str;=0A=
}=0A=
function removePref(name) {=0A=
	var prefArray =3D getPrefArray();=0A=
	name =3D convertPrefString(name);=0A=
	if (prefDefined(name)) {=0A=
		var newArray =3D new Array(prefArray.length - 1);=0A=
		var match =3D name + ":";=0A=
		var newArrIndex =3D 0;=0A=
		for (var i =3D 0; i < prefArray.length; i++) {=0A=
			if (!((prefArray[i].indexOf(match) =3D=3D 0) || (prefArray[i] =3D=3D =
name))) {=0A=
				newArray[newArrIndex++] =3D prefArray[i];=0A=
			}=0A=
		}=0A=
		setPrefCookie(newArray);=0A=
		if (prefArray.length =3D=3D 1) {=0A=
			return null;=0A=
		}=0A=
		else {=0A=
			return newArray;=0A=
		}=0A=
	}=0A=
	else {=0A=
		return prefArray;=0A=
	}=0A=
}=0A=
function setPref(name) {=0A=
	return setPref(name, null);=0A=
}=0A=
function setPref(name, value) {=0A=
	if (name) {=0A=
		name =3D convertPrefString(name);=0A=
		var prefArray =3D removePref(name);=0A=
		if (prefArray =3D=3D null) {=0A=
			prefArray =3D new Array;=0A=
		}=0A=
		var newArray =3D new Array(prefArray.length + 1);=0A=
		var newPref =3D name + (((value !=3D null) && (value !=3D '')) ? (':' =
+ value) : '');=0A=
		newArray[0] =3D newPref;=0A=
		for (var i =3D 0; i < prefArray.length; i++) {=0A=
			newArray[i+1] =3D prefArray[i];=0A=
		}=0A=
		setPrefCookie(newArray);=0A=
		return newArray;=0A=
	}=0A=
	else {=0A=
		return getPrefArray();=0A=
	}=0A=
}=0A=
function setPrefCookie(prefs) {=0A=
	if (prefs && (prefs.length > 0)) {=0A=
		var prefString;=0A=
		if (typeof(prefs) =3D=3D 'string') {=0A=
			prefString =3D prefs;=0A=
		}=0A=
		else {=0A=
			if (prefs.length =3D=3D 1) {=0A=
				prefString =3D prefs[0];=0A=
			}=0A=
			else {=0A=
				prefString =3D prefs.join(',');=0A=
			}=0A=
		}=0A=
		setCookie(UIPrefsCk.name, prefString, UIPrefsCk.expDays, =
UIPrefsCk.path, false, false);=0A=
	}=0A=
	else {=0A=
		deleteCookie(UIPrefsCk.name, UIPrefsCk.path, false);=0A=
	}=0A=
}=0A=
// individual pref items in the cookie are separated by ',' and=0A=
// nv pref pairs are joined by ':', i.e. n1:v1,n2:v2,n3:v3,etc.=0A=
// values are optional. (i.e. "n1,n2:v2,n3" is ok)=0A=
function getPrefArray() {=0A=
	var cookieVal =3D getCookie(UIPrefsCk.name);=0A=
	if (cookieVal !=3D null) {=0A=
		return cookieVal.split(',');=0A=
	}=0A=
	return null;=0A=
}=0A=
=0A=
// http://www.dustindiaz.com/top-ten-javascript/=0A=
function getCookie( name ) {=0A=
	var start =3D document.cookie.indexOf( name + "=3D" );=0A=
	var len =3D start + name.length + 1;=0A=
	if (((!start ) && (name !=3D document.cookie.substring( 0, name.length =
))) || (start =3D=3D -1)) {=0A=
		return null;=0A=
	}=0A=
	var end =3D document.cookie.indexOf( ';', len );=0A=
	if ( end =3D=3D -1 ) end =3D document.cookie.length;=0A=
	return unescape( document.cookie.substring( len, end ) );=0A=
}=0A=
function setCookie( name, value, expires, path, domain, secure ) {=0A=
	var today =3D new Date();=0A=
	today.setTime( today.getTime() );=0A=
	if ( expires ) {=0A=
		expires =3D expires * 1000 * 60 * 60 * 24;=0A=
	}=0A=
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------=_NextPart_000_0000_01C98BC5.4FC41B00
Content-Type: application/octet-stream
Content-Transfer-Encoding: quoted-printable
Content-Location: http://ajcp.ascpjournals.org/shared/js/design/hw-design1.js

$(document).ready(function() {=0A=
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defaultColorString, textColorString) {=0A=
	var inputSuggColor=3D"#A0A0A0"=0A=
	$("#hdr-login input").css("color",inputSuggColor);=0A=
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			if (mylabel.length && $(this).val() =3D=3D mylabel.text()) {=0A=
				$(this).val('').css("color","black");=0A=
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	);=0A=
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			if ($(this).val() =3D=3D '') {=0A=
				var myid =3D $(this).attr("id");=0A=
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					$(this).val(mylabel.text()).css("color",inputSuggColor);=0A=
				}=0A=
			}=0A=
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	);=0A=
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		function(e) {=0A=
			$("#hdr-login form").submit();=0A=
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	);=0A=
	=0A=
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------=_NextPart_000_0000_01C98BC5.4FC41B00
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	var gNavHeight =3D ((gNav.length) ? navOrigHeight : 0);=0A=
	var gNavTopOffset =3D  (gNavHeight > 0 ? gNav.offset().top : 0);=0A=
	var gScrollPos =3D 0;=0A=
	var gNavDocked =3D false;=0A=
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	var isIE6 =3D ($.browser.msie && ($.browser.version.substr(0,2) =3D=3D =
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	=0A=
	/* fig-expansion in page */=0A=
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	=0A=
	/* 'new window' fig expansions */=0A=
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}=0A=
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		error: ajaxErr,=0A=
		beforeSend: addFigHeaders,=0A=
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}=0A=
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=0A=
function addFig(xhtmlData, figWrapperEl) {=0A=
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lookupRule + ")", 1000);=0A=
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}=0A=
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=0A=
function dispRef(e) {=0A=
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		$("div#hovering-ref").remove();=0A=
		alert("hovering-ref div removed on new hover!");=0A=
	}	=0A=
	var newDiv =3D '<div id=3D"hovering-ref">' + =
($(link).next("div").children("div.cit-metadata").clone().html()) + =
'</div>';=0A=
	$("body").append(newDiv);=0A=
	var elH =3D getObjHeight($("div#hovering-ref"));=0A=
	$("div#hovering-ref").css("left", e.pageX+10).css("top", =
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=0A=
function hideRef(e) {=0A=
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		$("div#hovering-ref").remove();=0A=
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=0A=
function getHWCiting() {=0A=
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	if (citingA.length) {=0A=
		var newA =3D '<a id=3D"cb-loading-hw-cited" href=3D"#">Loading citing =
article data...</a>';=0A=
		citingA.replaceWith(newA);=0A=
		var href =3D citingA.attr("href");=0A=
		var id =3D '';=0A=
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					type: "GET",=0A=
					error: ajaxErr,=0A=
					success: addHWCiting,=0A=
					complete: ajaxComplete=0A=
				});=0A=
=0A=
			}=0A=
		}=0A=
	}=0A=
}=0A=
function getHWRelatedURLs() {=0A=
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			if (!(id =3D=3D '')) {=0A=
				var host =3D document.location.protocol + "//" + =
document.location.host;=0A=
				var ajaxUrl =3D host + '/related-web-pages/' + id.replace(/;/,'/');=0A=
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					url: ajaxUrl,=0A=
					dataType: "html",=0A=
					type: "GET",=0A=
					error: ajaxErr,=0A=
					success: addRelatedURLs,=0A=
					complete: ajaxComplete=0A=
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=0A=
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}=0A=
function getISIRelated() {=0A=
	var relatedA =3D $("#cb-isi-similar-articles");=0A=
	if (relatedA.length) {=0A=
		var newA =3D '<a id=3D"cb-isi-similar-articles" href=3D"#">Loading ISI =
article data...</a>';=0A=
		relatedA.replaceWith(newA);=0A=
		var href =3D relatedA.attr("href");=0A=
		var id =3D '';=0A=
		if (href) {=0A=
			var hrefDec =3D decodeURI(href);=0A=
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				var args =3D hrefDec.substring(hrefDec.indexOf('?') + 1).split('&');=0A=
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					var host =3D document.location.protocol + "//" + =
document.location.host;=0A=
					var ajaxUrl =3D host + '/isi-links/has-related/' + =
id.replace(/;/,'/');=0A=
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						url: ajaxUrl,=0A=
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						type: "GET",=0A=
						error: ajaxErr,=0A=
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						complete: ajaxComplete=0A=
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				}=0A=
			}=0A=
		}=0A=
	}=0A=
}=0A=
function getISICiting() {=0A=
	var citingA =3D $("#cb-isi-citing-articles");=0A=
	if (citingA.length) {=0A=
		var newA =3D '<a id=3D"cb-loading-isi-cited" href=3D"#">Loading ISI =
citing article data...</a>';=0A=
		citingA.replaceWith(newA);=0A=
		var href =3D citingA.attr("href");=0A=
		var id =3D '';=0A=
		if (href) {=0A=
			var hrefDec =3D decodeURI(href);=0A=
			if ((hrefDec.indexOf('?') > 0)) {=0A=
				var args =3D hrefDec.substring(hrefDec.indexOf('?') + 1).split('&');=0A=
				for (var i =3D 0; i < args.length; i++) {=0A=
					var argDec =3D decodeURIComponent(args[i]);=0A=
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					}=0A=
				}=0A=
				if (!(id =3D=3D '')) {=0A=
					var host =3D document.location.protocol + "//" + =
document.location.host;=0A=
					var ajaxUrl =3D host + '/isi-links/' + id.replace(/;/,'/');=0A=
					$.ajax({=0A=
						url: ajaxUrl,=0A=
						dataType: "html",=0A=
						type: "GET",=0A=
						error: ajaxErr,=0A=
						success: addISICiting,=0A=
						complete: ajaxComplete=0A=
					});=0A=
				}=0A=
			}=0A=
		}=0A=
	}=0A=
}=0A=
function getEntrezLinks() {=0A=
	var entrezDiv =3D $("#cb-entrez-links-placeholder");=0A=
	if (entrezDiv.length) {=0A=
		var entrezA =3D entrezDiv.children("a");=0A=
		if (entrezA) {=0A=
			var host =3D document.location.protocol + "//" + =
document.location.host;=0A=
			var ajaxUrl =3D host + entrezA.attr("href");=0A=
			$.ajax({=0A=
				url: ajaxUrl,=0A=
				dataType: "html",=0A=
				type: "GET",=0A=
				error: ajaxErr,=0A=
				success: addEntrezLinks,=0A=
				complete: ajaxComplete=0A=
			});=0A=
		}=0A=
	}=0A=
}=0A=
=0A=
function ajaxErr(req, msg, e) {=0A=
}=0A=
function ajaxComplete(req, msg) {=0A=
}=0A=
function updateCBItem(cbItem, newHTML, hasData) {=0A=
	var parentItem =3D cbItem.parents("li").eq(0);=0A=
	cbItem.replaceWith(newHTML);=0A=
	if (!hasData) {=0A=
		// hide the parent li=0A=
		if (parentItem.length) {=0A=
			modClass(parentItem,"nodata","");=0A=
			// check if there are any siblings still being displayed=0A=
			var otherItems =3D parentItem.siblings();=0A=
			var allItemsEmpty;=0A=
			if (otherItems.length) {=0A=
				if (otherItems.length =3D=3D otherItems.filter(".nodata").length) {=0A=
					allItemsEmpty =3D true=0A=
				}=0A=
				else {=0A=
					allItemsEmpty =3D false;=0A=
				}=0A=
			}=0A=
			else {=0A=
				allItemsEmpty =3D true;=0A=
			}=0A=
			if (allItemsEmpty) {=0A=
				var cbsection =3D parentItem.parents("div.cb-section").eq(0);=0A=
				if (cbsection.length) {=0A=
					modClass(cbsection,"nodata","");=0A=
				}=0A=
			}=0A=
		}=0A=
	}=0A=
	// in frameset fix targets on child links, forms=0A=
	fixFrameLinks(parentItem.find("a,form"));=0A=
	fixColHeights(2);=0A=
}=0A=
function fixFrameLinks(jqItems) {=0A=
	if ((gIsFrameset !=3D null) && gIsFrameset) {=0A=
		jqItems.each(=0A=
			function(i) {=0A=
				$(this).attr("target", "_top");=0A=
			}=0A=
		);=0A=
	}=0A=
}=0A=
function addRelatedURLs(xhtmlData) {=0A=
	var cbA =3D $("#cb-loading-related-urls");=0A=
	if (xhtmlData && !(xhtmlData.indexOf('<span') >=3D 0)) {=0A=
		$("#related-urls").replaceWith(xhtmlData);=0A=
		fixColHeights(1);=0A=
		if (cbA.length) {=0A=
			updateCBItem(cbA, '<a href=3D"#related-urls">Related Web Pages</a>', =
true);=0A=
		}=0A=
	}=0A=
	else {=0A=
		if (cbA.length) {=0A=
			updateCBItem(cbA, '<div id=3D"cb-loaded-related-urls-none">No related =
web pages</div>', false);=0A=
		}=0A=
	}=0A=
}=0A=
function addHWCiting(xhtmlData) {=0A=
	var cbA =3D $("#cb-loading-hw-cited");=0A=
	if (xhtmlData) {=0A=
		$("#content-block").append(xhtmlData);=0A=
		fixColHeights(1);=0A=
		if (cbA.length) {=0A=
			updateCBItem(cbA, '<a href=3D"#cited-by">View citing article =
information</a>', true);=0A=
		}=0A=
	}=0A=
	else {=0A=
		if (cbA.length) {=0A=
			updateCBItem(cbA, '<div id=3D"cb-loaded-hw-cited-none">No citing =
articles</div>', false);=0A=
		}=0A=
	}=0A=
}=0A=
function addISIRelated(xhtmlData) {=0A=
	var cbA =3D $("#cb-isi-similar-articles");=0A=
	if (xhtmlData && !(xhtmlData.indexOf('<span') >=3D 0)) {=0A=
		if (cbA.length) {=0A=
			updateCBItem(cbA, xhtmlData, true);=0A=
		}=0A=
	}=0A=
	else {=0A=
		if (cbA.length) {=0A=
			updateCBItem(cbA, '<div id=3D"cb-loaded-isi-related-none">No ISI =
related articles</div>', false);=0A=
		}=0A=
	}=0A=
}=0A=
function addISICiting(xhtmlData) {=0A=
	var cbA =3D $("#cb-loading-isi-cited");=0A=
	if (xhtmlData && !(xhtmlData.indexOf('<span') >=3D 0)) {=0A=
		if (cbA.length) {=0A=
			updateCBItem(cbA, xhtmlData, true);=0A=
		}=0A=
	}=0A=
	else {=0A=
		if (cbA.length) {=0A=
			updateCBItem(cbA, '<div id=3D"cb-loaded-isi-cited-none">No ISI citing =
articles</div>', false);=0A=
		}=0A=
	}=0A=
}=0A=
function addEntrezLinks(xhtmlData) {=0A=
	var entrezDiv =3D $("#cb-entrez-links-placeholder");=0A=
	if (xhtmlData) {=0A=
		entrezDiv.empty().append(xhtmlData);=0A=
		fixFrameLinks(entrezDiv.find("a,form"));=0A=
		fixColHeights(2);=0A=
	}=0A=
}=0A=
function addDockedNav() {=0A=
	var slugEl =3D $("#slugline");=0A=
	// get direct children li elements only=0A=
	var artViews =3D $("#article-cb-main .cb-section ol:has(li =
a[rel^=3D'view-']) > li").clone();=0A=
	var newDiv =3D '<div id=3D"docked-nav"></div>';=0A=
	$("#col-2").append(newDiv);=0A=
	$("#col-2 #docked-nav").hide();=0A=
	var newDivJQuery =3D $("#docked-nav");=0A=
	newDivJQuery.append('<div class=3D"content-box"><div =
class=3D"cb-contents"><h3>This Article</h3><div class=3D"cb-section =
cb-slug"><ol id=3D"docked-slug"><li></li></ol></div><div =
class=3D"cb-section"><ol =
id=3D"docked-nav-views"></ol></div></div></div>');=0A=
	$("#col-2 #docked-nav-views").append(artViews); /*.append(artSupp);*/=0A=
	$("#col-2 #docked-nav #docked-slug li").append(slugEl.clone());=0A=
	newDivJQuery.append($("#article-dyn-nav").clone());=0A=
	$("#col-2 #docked-nav").fadeIn(250);=0A=
}=0A=
=0A=
=0A=
function removeDockedNav() {=0A=
	var dockedNav =3D $("div#docked-nav");=0A=
	if(dockedNav.length) {=0A=
		dockedNav.fadeOut(250, function() { dockedNav.remove(); });=0A=
	}=0A=
}=0A=

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