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Subject: PLoS ONE: Mutations in H5N1 Influenza Virus Hemagglutinin that Confer Binding to Human Tracheal Airway Epithelium
Date: Thu, 19 Nov 2009 16:34:14 -0800
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<H1 property=3D"dc:title" xpathLocation=3D"noSelect"=20
href=3D"http://purl.org/dc/dcmitype/Text" rel=3D"dc:type" =
datatype=3D"">Mutations in=20
H5N1 Influenza Virus Hemagglutinin that Confer Binding to Human Tracheal =
Airway=20
Epithelium</H1>
<DIV style=3D"DISPLAY: none; VISIBILITY: hidden" =
property=3D"dc:description"=20
datatype=3D"">
<P>The emergence in 2009 of a swine-origin H1N1 influenza virus as the =
first=20
pandemic of the 21st Century is a timely reminder of the international =
public=20
health impact of influenza viruses, even those associated with mild =
disease. The=20
widespread distribution of highly pathogenic H5N1 influenza virus in the =
avian=20
population has spawned concern that it may give rise to a human =
influenza=20
pandemic. The mortality rate associated with occasional human infection =
by H5N1=20
virus approximates 60%, suggesting that an H5N1 pandemic would be =
devastating to=20
global health and economy. To date, the H5N1 virus has not acquired the=20
propensity to transmit efficiently between humans. The reasons behind =
this are=20
unclear, especially given the high mutation rate associated with =
influenza virus=20
replication. Here we used a panel of recombinant H5 hemagglutinin (HA) =
variants=20
to demonstrate the potential for H5 HA to bind human airway epithelium, =
the=20
predominant target tissue for influenza virus infection and spread. =
While=20
parental H5 HA exhibited limited binding to human tracheal epithelium,=20
introduction of selected mutations converted the binding profile to that =
of a=20
current human influenza strain HA. Strikingly, these amino-acid changes =
required=20
multiple simultaneous mutations in the genomes of naturally occurring H5 =

isolates. Moreover, H5 HAs bearing intermediate sequences failed to bind =
airway=20
tissues and likely represent mutations that are an evolutionary =
=E2=80=9Cdead end.=E2=80=9D We=20
conclude that, although genetic changes that adapt H5 to human airways =
can be=20
demonstrated, they may not readily arise during natural virus =
replication. This=20
genetic barrier limits the likelihood that current H5 viruses will =
originate a=20
human pandemic.</P></DIV><SPAN content=3D"2009-11-18" =
property=3D"dc:date"=20
href=3D"http://dx.doi.org/10.1371/journal.pone.0007836" =
rel=3D"dc:identifier"=20
datatype=3D"xsd:date"></SPAN><SPAN content=3D"Infectious Diseases"=20
property=3D"dc:subject"></SPAN><SPAN content=3D"Respiratory Infections"=20
property=3D"dc:subject"></SPAN><SPAN content=3D"Respiratory Medicine"=20
property=3D"dc:subject"></SPAN><SPAN content=3D"Viral Infections"=20
property=3D"dc:subject"></SPAN><SPAN content=3D"Emerging Viral Diseases" =

property=3D"dc:subject"></SPAN><SPAN content=3D"Host Invasion and Cell =
Entry"=20
property=3D"dc:subject"></SPAN><SPAN content=3D"Virology"=20
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<P class=3Dauthors xpathLocation=3D"noSelect"=20
xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema"><SPAN=20
property=3D"dc:creator">Guadalupe Ayora-Talavera</SPAN><SUP><A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#aff1">1</A></SUP><SUP><A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#equal-contrib">#</A></SUP>,=20
<SPAN property=3D"dc:creator">Holly Shelton</SPAN><SUP><A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#aff1">1</A></SUP><SUP><A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#equal-contrib">#</A></SUP>,=20
<SPAN property=3D"dc:creator">Margaret A. Scull</SPAN><SUP><A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#aff2">2</A></SUP>,=20
<SPAN property=3D"dc:creator">Junyuan Ren</SPAN><SUP><A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#aff3">3</A></SUP>,=20
<SPAN property=3D"dc:creator">Ian M. Jones</SPAN><SUP><A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#aff3">3</A></SUP>,=20
<SPAN property=3D"dc:creator">Raymond J. Pickles</SPAN><SUP><A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#aff2">2</A></SUP>,=20
<SPAN property=3D"dc:creator">Wendy S. Barclay</SPAN><SUP><A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#aff1">1</A></SUP><SUP><A=20
class=3Dfnoteref=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#cor1">*</A></SUP></P>
<P class=3Daffiliations xpathLocation=3D"noSelect"=20
xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema"><A id=3Daff1=20
name=3Daff1></A><STRONG>1</STRONG> Department of Virology, Imperial =
College=20
London, London, United Kingdom, <A id=3Daff2 =
name=3Daff2></A><STRONG>2</STRONG>=20
Cystic Fibrosis/Pulmonary Research and Treatment Center, Department of=20
Microbiology and Immunology, University of North Carolina at Chapel =
Hill, Chapel=20
Hill, North Carolina, United States of America, <A id=3Daff3=20
name=3Daff3></A><STRONG>3</STRONG> School of Biological Sciences, =
University of=20
Reading, Whiteknights, Reading, United Kingdom</P>
<DIV class=3Dabstract=20
xpathLocation=3D"/article[1]/front[1]/article-meta[1]/abstract[1]"=20
xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema"><A id=3Dabstract0 =
title=3DAbstract=20
name=3Dabstract0 toc=3D"abstract0"></A>
<H2 xpathLocation=3D"noSelect">Abstract&nbsp;<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#top">Top</A></H2>
<P =
xpathLocation=3D"/article[1]/front[1]/article-meta[1]/abstract[1]/p[1]">T=
he=20
emergence in 2009 of a swine-origin H1N1 influenza virus as the first =
pandemic=20
of the 21st Century is a timely reminder of the international public =
health=20
impact of influenza viruses, even those associated with mild disease. =
The=20
widespread distribution of highly pathogenic H5N1 influenza virus in the =
avian=20
population has spawned concern that it may give rise to a human =
influenza=20
pandemic. The mortality rate associated with occasional human infection =
by H5N1=20
virus approximates 60%, suggesting that an H5N1 pandemic would be =
devastating to=20
global health and economy. To date, the H5N1 virus has not acquired the=20
propensity to transmit efficiently between humans. The reasons behind =
this are=20
unclear, especially given the high mutation rate associated with =
influenza virus=20
replication. Here we used a panel of recombinant H5 hemagglutinin (HA) =
variants=20
to demonstrate the potential for H5 HA to bind human airway epithelium, =
the=20
predominant target tissue for influenza virus infection and spread. =
While=20
parental H5 HA exhibited limited binding to human tracheal epithelium,=20
introduction of selected mutations converted the binding profile to that =
of a=20
current human influenza strain HA. Strikingly, these amino-acid changes =
required=20
multiple simultaneous mutations in the genomes of naturally occurring H5 =

isolates. Moreover, H5 HAs bearing intermediate sequences failed to bind =
airway=20
tissues and likely represent mutations that are an evolutionary =
=E2=80=9Cdead end.=E2=80=9D We=20
conclude that, although genetic changes that adapt H5 to human airways =
can be=20
demonstrated, they may not readily arise during natural virus =
replication. This=20
genetic barrier limits the likelihood that current H5 viruses will =
originate a=20
human pandemic.</P></DIV>
<DIV class=3Darticleinfo xpathLocation=3D"noSelect"=20
xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema">
<P><STRONG>Citation: </STRONG>Ayora-Talavera G, Shelton H, Scull MA, Ren =
J,=20
Jones IM, et al. (2009) Mutations in H5N1 Influenza Virus Hemagglutinin =
that=20
Confer Binding to Human Tracheal Airway Epithelium. PLoS ONE 4(11): =
e7836.=20
doi:10.1371/journal.pone.0007836</P>
<P><STRONG>Editor: </STRONG>Cheryl A. Stoddart, University of California =
San=20
Francisco, United States of America</P>
<P></P>
<P><STRONG>Received:</STRONG> April 16, 2009; <STRONG>Accepted:</STRONG> =
October=20
13, 2009; <STRONG>Published:</STRONG> November 18, 2009</P>
<P><STRONG>Copyright:</STRONG> =C2=A9 2009 Ayora-Talavera et al. This is =
an=20
open-access article distributed under the terms of the Creative Commons=20
Attribution License, which permits unrestricted use, distribution, and=20
reproduction in any medium, provided the original author and source are=20
credited.</P>
<P><STRONG>Funding:</STRONG> This work was supported by Medical Research =
Council=20
award G0500597 to I. M. Jones and W. S. Barclay and by the National =
Institutes=20
of Health (NIH) Molecular Biology of Viral Diseases Training Grant=20
5-T32-AI007419, NIH R01 HL77844-1, NIH R21 HL080098-01, SCCOR 5 P50 =
HL084934=20
(RJP). G. Ayora-Talvera was funded by PROMEP-Secretaria de Educacion =
Publica,=20
Mexico. Margaret Scull (nee Hennessey) is a recipient of the George H. =
Hitchings=20
Fund for Health Research and Science Education of the Triangle Community =

Foundation. W.S. Barclay is a member of the Wellcome Trust-funded Centre =
for=20
Respiratory Infection Research. The funders had no role in study design, =
data=20
collection and analysis, decision to publish, or preparation of the=20
manuscript.</P>
<P><STRONG>Competing interests:</STRONG> The authors have declared that =
no=20
competing interests exist.</P>
<P><A name=3Dcor1></A>* E-mail: <A=20
href=3D"mailto:w.barclay@imperial.ac.uk">w.barclay@imperial.ac.uk</A></P>=

<P><A name=3Dequal-contrib></A># These authors contributed equally to =
this work.=20
</P></DIV>
<DIV id=3Dsection1 xpathLocation=3D"/article[1]/body[1]/sec[1]"=20
xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema"><A id=3Ds1 =
title=3DIntroduction name=3Ds1=20
toc=3D"s1"></A>
<H3 xpathLocation=3D"noSelect">Introduction&nbsp;<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#top">Top</A></H3>
<P xpathLocation=3D"/article[1]/body[1]/sec[1]/p[1]">Influenza pandemics =
occur=20
periodically and their significant effect on society is aptly =
demonstrated by=20
the current ongoing pandemic caused by a swine origin H1N1 subtype =
influenza=20
virus that has thus far claimed nearly 2000 lives in the span of 6 =
months in mid=20
2009 <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-WHO1">[1]</A>.=20
Avian influenza H5N1 viruses have infected more than 400 people since =
2003 but=20
no sustained human-to-human transmission has occurred to date <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-WHO2">[2]</A>.=20
The reasons why H5N1 fails to transmit among humans remain unclear =
especially=20
since high viral titres have been isolated from the nose and throat of =
H5N1=20
infected individuals <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-deJong1">[3]</A>.=20
The host barrier for avian influenza virus spread in humans is =
multigenic. Many=20
H5N1 isolates already contain mutations in viral genes such as the PB2 =
component=20
of the polymerase complex that confer enhanced replication capacity in =
human=20
cells, but still this has not rendered them transmissible. Other viral =
genes=20
such as the viral hemagglutintin protein (HA) are undoubtedly key to=20
understanding the H5N1 virus' pandemic potential.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[1]/p[2]">The viral =
hemagglutinin=20
protein (HA) in part dictates host range and cellular tropism of =
influenza virus=20
through its preference for different conformers of sialic acid (SA) on =
host cell=20
glycans that mediate virus attachment and facilitate infection (reviewed =
by <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Kuiken1">[4]</A>).=20
In previous studies, the distribution of SA in the human airway =
epithelium has=20
been investigated using linkage-specific lectins as SA probes. These =
studies=20
showed =CE=B12-6 SA expression was abundant in the upper respiratory =
tract whereas=20
=CE=B12-3 SA was scarce <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-vanRiel1">[5]</A>.=20
In contrast, in the distal lung regions both =CE=B12-3 and 2=E2=80=936 =
SA expression was=20
significant <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Gagneux1">[6]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Shinya1">[7]</A>.=20
While the precise nature of the molecule(s) used by influenza virus as a =

receptor are not known, sialic acid residues in the =CE=B12-6 conformer =
are=20
preferentially used by the HA proteins of human influenza viruses while =
avian=20
isolate HAs prefer SA with =CE=B12-3 linkages <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Matrosovich1">[8]</A>.=20
Significant evidence exists that a switch in SA preference of the HA, =
from =CE=B12-3=20
to =CE=B12-6 linked SA, is a necessary adaptation step for viruses with =
avian HAs to=20
gain the ability for human-to-human transmission. Previous influenza =
pandemics=20
of 1918, 1957 and 1968 support this requirement as each of the =
respective HA=20
genes, originating from avian-like viruses, mutated to increase binding=20
preference for =CE=B12-6 SA <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Matrosovich1">[8]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Tumpey1">[9]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Reid1">[10]</A>.=20
Notably two amino acid mutations in the HA receptor binding site (RBS) =
were=20
important during the evolution of each pandemic virus. Viral sequences =
from=20
early isolates in the pandemics suggest adaptation occurred in a =
stepwise manner=20
as isolates with just one of the two mutations have occasionally been =
found <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Reid1">[10]</A>.=20
Further, transmissibility of the 1918 influenza virus between ferrets =
was lost=20
after conversion of two amino acids in the receptor binding site of HA =
to those=20
of avian H1N1 viruses, whereas mutation of a single amino acid produced =
a viable=20
virus with intermediate transmissibility <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Tumpey1">[9]</A>.=20
Recovered sequences from other sources of 1918 influenza viruses =
indicate that=20
the intermediate mutations also circulated during that time <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Reid1">[10]</A>.=20
Whether or not the current H5N1 viruses are capable of progressive =
mutations=20
that switch their SA linkage preference is central to an accurate risk=20
assessment of the potential of H5N1 to be the progenitor of a human =
influenza=20
virus pandemic.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[1]/p[3]">A wide range of =
mutations=20
that switch receptor preference of HA proteins have been identified and =
include=20
changes throughout the HA gene. Key amino acids at positions 226 and 228 =
(H3 HA=20
numbering) affect SA preference of several subtypes including H2, H3, H4 =
and H9=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Matrosovich1">[8]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Bateman1">[11]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Wan1">[12]</A>.=20
On the other hand the H1 HAs that are human adapted bear changes at =
positions=20
190 and 225 relative to their avian precursors <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Tumpey1">[9]</A>.=20
Mutations of the H3 HA at position 190, 193 and 194 were all previously =
shown to=20
alter SA preference <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Ibricevic1">[13]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Medeiros1">[14]</A>.=20
Changes in HA that affect the extent of its glycosylation can also =
profoundly=20
affect the SA binding affinity and in vivo this may contribute to =
changes in=20
tropism and pathogenicity <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Thompson1">[15]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Owen1">[16]</A>.=20
A selection of characterized mutations were previously engineered into=20
recombinant H5 HA proteins and their effects on SA binding studied with =
red=20
blood cells from different species <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Yang1">[17]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Harvey1">[18]</A>=20
or using synthetic glycans which act as receptor mimetics <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Ilyushina1">[19]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Stevens1">[20]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Stevens2">[21]</A>.=20
None of these mutations when present in H5 HA conferred a switch from =
=CE=B12-3 SA=20
binding to =CE=B12-6 SA binding using these assays. However, SA =
presentation on red=20
blood cells or as glycan mimetics may not accurately represent SA =
expressed on=20
the human airway epithelium luminal surface. In addition, the use of =
lectins as=20
probes for SA that are utilized by influenza viruses has recently been=20
questioned <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Qi1">[22]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Nicholls1">[23]</A>.=20
Although lectins are highly specific for terminal linkages in =
glycopolymer=20
chains they do not interact with adjacent carbohydrates. Conversely, =
influenza=20
HA proteins have more complex interactions with sialylated glycans and =
binding=20
of HA to SA are affected by glycan topology at the cell surface <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Chandrasekaran1">[24]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Srinivasan1">[25]</A>.=20
Thus, lectins may detect SA that cannot be utilized as receptors for =
influenza=20
viruses and HA proteins may bind more avidly to some sialylated =
receptors not=20
readily detected by lectins. Tests to assess binding of H5 and mutants =
thereof=20
in relation to host range receptor switching should therefore strive to =
use=20
models that reflect the microenvironment of the luminal surface of the =
human=20
airway epithelium.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[1]/p[4]">The human airway =
epithelium=20
is a well-differentiated tissue consisting of ciliated cells and =
mucin-secreting=20
cells that overlay a basal cell layer. <I>In vitro</I> models of human =
ciliated=20
airway epithelium (HAE) that recapitulates the morphology, biochemistry =
and=20
physiology of the epithelium are available. Such models have been used =
to study=20
virus-host cell interactions for a number of respiratory virus including =
human=20
and avian influenza viruses <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Thompson1">[15]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Bartlett1">[26]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Zhang1">[27]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Zhang2">[28]</A>.=20
In particular, we and others have shown that human and avian influenza =
viruses=20
target different cell types in this model, a tropism related to the =
specific SA=20
linkage displayed on these cells <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Thompson1">[15]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Matrosovich2">[29]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Wan2">[30]</A>.=20
Recent data also suggest that the differential tropism in HAE cultures =
may=20
account for the compromised spread of avian influenza viruses in the =
cooler=20
conditions of the human upper respiratory tract<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Scull1">[31]</A>.=20
In this model, avian influenza viruses exclusively infect ciliated cells =
whereas=20
human influenza viruses that showed a switch in receptor preference to=20
predominately =CE=B12-6 SA displayed an expanded cell tropism where both =
ciliated=20
cells and non-ciliated columnar cells were infected. In addition, =
preferential=20
binding of the H1 HA of the 1918 human influenza virus to non-ciliated =
cells of=20
human tracheal tissue sections was recently reported <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Srinivasan1">[25]</A>.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[1]/p[5]">Here, we use HAE =
cultures and=20
tissue sections of human tracheal epithelium to determine the binding=20
characteristics of wild-type and select mutations of H5 HA to human =
airway=20
epithelium. In contrast to results published with short synthetic =
sialylated=20
ligands, we show that mutations in H5 HA at residue 228 effectively =
switch the=20
binding pattern from an avian-like HA to that shown by the HA of a human =

influenza H3 virus. Detailed sequence analysis of codon usage at this =
site by H5=20
revealed that although this mutation is possible, currently circulating =
H5N1=20
viruses are not poised to acquire this amino acid change easily. Our =
findings=20
confirm the potential for H5 to adapt to human airway epithelial cell =
receptors=20
but suggest that it is unlikely to occur within the quasi-species =
present in an=20
infected individual. Our data offers a rationale for why H5N1 viruses =
have not=20
yet evolved into human transmissible strains with pandemic =
potential.</P></DIV>
<DIV id=3Dsection2 xpathLocation=3D"/article[1]/body[1]/sec[2]"=20
xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema"><A id=3Ds2 title=3DResults =
name=3Ds2=20
toc=3D"s2"></A>
<H3 xpathLocation=3D"noSelect">Results&nbsp;<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#top">Top</A></H3>
<H4 =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/title[1]">Expression =
and=20
glycan binding of synthetic H5 HA proteins</H4>
<P xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/p[1]">We generated =

recombinant wild-type and mutant H5 HAs and used them as probes to =
determine=20
their ability to bind to SA on the luminal surface of human airway =
epithelium.=20
For our wild-type H5 HA we chose the A/Vietnam/1194/04 H5N1 isolate =
(accession=20
number: AY651333), as this virus was isolated from an infected human =
lung but=20
was not associated with human-to-human transmission. Wild-type H5 HA was =
cloned=20
as a fusion protein with the Fc domain of human immunoglobulin heavy =
chain as a=20
immunological tag and expressed as a soluble molecule in the supernatant =
of=20
recombinant baculovirus-infected insect cells (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g001">Figure=20
1A</A> and <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Barclay1">[32]</A>).=20
We then introduced either single or double mutations at Q226L and G228S =
in the=20
SA binding site of H5 HA <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Stevens2">[21]</A>=20
equivalent to those previously shown to switch receptor specificity in =
H2 and H3=20
HA subtypes <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Matrosovich1">[8]</A>.=20
Our previous work has shown these mutations alter red blood cell binding =
by H5=20
HA <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Harvey1">[18]</A>.=20
Equivalent mutations have not been documented in H5N1 viruses isolated =
in=20
nature. For comparison, we also generated a S227N mutant identified in =
several=20
human H5N1 virus isolates and reported to show changes in SA binding =
profiles in=20
synthetic glycan assays <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Gambaryan1">[33]</A>.=20
All HA mutants were similarly expressed using recombinant baculoviruses =
and were=20
secreted from infected <I>Sf</I>9 cells at similar level to the =
wild-type H5 HA=20
protein <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Barclay1">[32]</A>=20
(<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g001">Figure=20
1B</A>). A solid phase binding assay was used to evaluate the binding of =
the=20
HA-Fc proteins to short synthetic receptor mimics bearing a single copy =
of=20
lactosamine with terminal SA in the =CE=B12-3 (3SLN) and =CE=B12-6 =
linkage (6SLN) (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g002">Figure=20
2</A>) <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Barclay1">[32]</A>.=20
As expected, H5 HA bound to 3SLN and not to 6SLN whereas H3 HA cloned =
from the=20
recently circulating human H3N2 virus (A/Panama/2007/99, accession =
number:=20
CY034100) showed the opposite preference. Most of the H5 HA mutants =
retained=20
affinity for the 3SLN glycopolymer. Interestingly, mutant Q226L did not =
bind to=20
either of the sialyated lactosamine ligands tested in this assay. We =
therefore=20
assayed Q226L H5 HA mutant protein for binding to the conventional =
=CE=B12-3SA=20
lactose ligand, 3SL, since this has been reported by others <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Gambaryan1">[33]</A>.=20
In agreement with the previous report, we found that both wild type H5 =
HA=20
protein and the Q226L mutant bound efficiently to the 3SL ligand =
confirming that=20
the Q226L protein was correctly folded and represented a relevant ligand =
for our=20
further studies. Of all the H5 HA mutants tested, only S227N acquired =
affinity=20
for 6SLN in accordance with previous findings <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Gambaryan1">[33]</A>.=20
This mutant also retained 3SLN affinity, although it was reduced =
compared to=20
that of the wild-type H5 HA.</P>
<DIV class=3Dfigure =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/fig[1]"><A=20
id=3Dpone-0007836-g001 title=3D"Click for larger image "=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g001"=20
name=3Dpone-0007836-g001><IMG class=3Dthumbnail border=3D1 =
alt=3Dthumbnail align=3Dleft=20
src=3D"http://www.plosone.org/article/fetchObject.action?uri=3Dinfo:doi/1=
0.1371/journal.pone.0007836.g001&amp;representation=3DPNG_S"=20
xpathLocation=3D"noSelect"></A>
<P><STRONG =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/fig[1]/label[1]"><A=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g001"><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/fig[1]/label[1]">Figur=
e 1.=20
</SPAN></A><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/fig[1]/caption[1]/titl=
e[1]">Expression=20
of recombinant HA-Fc proteins.</SPAN></STRONG></P>
<P =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/fig[1]/caption[1]/p[1]=
">(A)=20
Recombinant baculoviruses encoding HA from a recent H3N2 human virus,=20
(A/Panama/2007/99) or from a highly pathogenic H5N1 avian influenza =
virus=20
(A/Vietnam/1194/04), were generated as previously described <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Barclay1">[32]</A>.=20
The HA proteins were expressed as soluble proteins secreted from =
infected=20
<I>Sf9</I> cells by removing the HA transmembrane (TM) and cytoplasmic =
tail (CT)=20
portions of the protein and replacing the HA signal peptide (SP) with =
the signal=20
peptide of the baculovirus envelope protein gp64 (SP gp64). The HA =
proteins were=20
tagged at the C-terminus by a human Fc (HuFc) and hexa-histidine (His6) =
tags.=20
(B) All recombinant HA proteins were expressed at similar levels as =
determined=20
by western blot analysis using an anti-human Fc antibody.</P><SPAN=20
xpathLocation=3D"noSelect">doi:10.1371/journal.pone.0007836.g001</SPAN>
<DIV class=3Dclearer></DIV></DIV>
<DIV class=3Dfigure =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/fig[2]"><A=20
id=3Dpone-0007836-g002 title=3D"Click for larger image "=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g002"=20
name=3Dpone-0007836-g002><IMG class=3Dthumbnail border=3D1 =
alt=3Dthumbnail align=3Dleft=20
src=3D"http://www.plosone.org/article/fetchObject.action?uri=3Dinfo:doi/1=
0.1371/journal.pone.0007836.g002&amp;representation=3DPNG_S"=20
xpathLocation=3D"noSelect"></A>
<P><STRONG =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/fig[2]/label[1]"><A=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g002"><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/fig[2]/label[1]">Figur=
e 2.=20
</SPAN></A><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/fig[2]/caption[1]/titl=
e[1]">Binding=20
of recombinant HA proteins to synthetic receptor ligands 3SLN (avian =
receptor)=20
and 6SLN (human receptor) in a solid phase assay.</SPAN></STRONG></P>
<P =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[1]/fig[2]/caption[1]/p[1]=
">(A)=20
Synthetic glycans on polyacrylamide linkers were immobilized on 96-well =
plates=20
following UV treatment. Recombinant HA proteins were preformed into =
higher order=20
complexes by incubation with anti-human Fc before incubation on the =
plate and=20
detection with goat anti-human IgG conjugated to horse-radish peroxidise =
(HRP).=20
(B) Recombinant HA-Fc proteins were adsorbed on 96 well plates coated =
with=20
anti-human Fc antibody. Synthetic glycans on polyacrylamide linkers with =
biotin=20
tags (6SLN and 3SL) were incubated on the plates and detected with =
Streptavidin=20
=E2=80=93 HRP conjugate.</P><SPAN=20
xpathLocation=3D"noSelect">doi:10.1371/journal.pone.0007836.g002</SPAN>
<DIV class=3Dclearer></DIV></DIV>
<H4 xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/title[1]">Binding =
of HA=20
proteins to an in vitro model of human airway epithelium</H4>
<P xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/p[1]">The human =
airway=20
epithelium, the predominant site of human influenza virus infection, =
contains a=20
mixture of ciliated and non-ciliated epithelial cell types. Ciliated =
cells are=20
abundant and can be identified morphologically by the presence of cilia =
and=20
immunologically by antibodies directed against their expression of =
acetylated=20
=CE=B1-tubulin. Amongst the non-ciliated epithelial cell populations, =
pre-ciliated=20
cells, mucin-secreting (Goblet cells), and other secretory cells may be =
present=20
depending on the anatomical site examined (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g003">Figure=20
3A</A> and <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g004">Figure=20
4A</A>). We and others have shown that using <I>in vitro</I> models of =
human=20
ciliated airway epithelium (HAE), human influenza viruses infect both =
ciliated=20
and non-ciliated cells, while avian influenza viruses infect only =
ciliated cells=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Thompson1">[15]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Matrosovich3">[34]</A>.=20
In HAE, =CE=B12-3 SA expression detected by MAA lectin binding was =
evident at the=20
tips of cilia and on the apical surface of ciliated cells likely at the =
level of=20
the microvilli. MAA binding was only rarely seen on cells that lacked =
obvious=20
cilia. In contrast, SNA lectin binding detected =CE=B12-6 SA on the =
apical surface of=20
both ciliated and non-ciliated cells <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Thompson1">[15]</A>.</P>
<DIV class=3Dfigure =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/fig[1]"><A=20
id=3Dpone-0007836-g003 title=3D"Click for larger image "=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g003"=20
name=3Dpone-0007836-g003><IMG class=3Dthumbnail border=3D1 =
alt=3Dthumbnail align=3Dleft=20
src=3D"http://www.plosone.org/article/fetchObject.action?uri=3Dinfo:doi/1=
0.1371/journal.pone.0007836.g003&amp;representation=3DPNG_S"=20
xpathLocation=3D"noSelect"></A>
<P><STRONG =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/fig[1]/label[1]"><A=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g003"><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/fig[1]/label[1]">Figur=
e 3.=20
</SPAN></A><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/fig[1]/caption[1]/titl=
e[1]">Recombinant=20
HA proteins bind to human airway epithelial (HAE) =
cultures.</SPAN></STRONG></P>
<P =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/fig[1]/caption[1]/p[1]=
">(A)=20
Morphological features of HAE cultures visualized by H&amp;E =
counterstain. Cell=20
types present include ciliated cells (C), mucin-secreting cells (MS) and =

non-ciliated cells (NC). HAE cultures were probed with recombinant HA =
proteins=20
from human H3 (A/Panama/2007/99) (B,C) or avian H5 (A/Vietnam/1194/04) =
(D,E)=20
viruses either directly to the apical surface of fixed cultures (B,D) or =
to=20
histological sections (C,E). Receptor binding site mutants of H5 HA =
(F=E2=80=93J) were=20
also analysed for binding to histological sections. Ciliated cells were=20
identified using anti acetylated =CE=B1-tubulin (green) and the HA-Fc =
proteins were=20
visualized with anti human-Fc (red). Images are representative of =
cell-type=20
binding seen in experiments. (K) Numbers and types of epithelial cells =
bound by=20
HA proteins were quantified by counting 100=E2=80=93200 total cells from =
five different=20
fields of view.</P><SPAN=20
xpathLocation=3D"noSelect">doi:10.1371/journal.pone.0007836.g003</SPAN>
<DIV class=3Dclearer></DIV></DIV>
<DIV class=3Dfigure =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/fig[2]"><A=20
id=3Dpone-0007836-g004 title=3D"Click for larger image "=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g004"=20
name=3Dpone-0007836-g004><IMG class=3Dthumbnail border=3D1 =
alt=3Dthumbnail align=3Dleft=20
src=3D"http://www.plosone.org/article/fetchObject.action?uri=3Dinfo:doi/1=
0.1371/journal.pone.0007836.g004&amp;representation=3DPNG_S"=20
xpathLocation=3D"noSelect"></A>
<P><STRONG =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/fig[2]/label[1]"><A=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g004"><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/fig[2]/label[1]">Figur=
e 4.=20
</SPAN></A><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/fig[2]/caption[1]/titl=
e[1]">Recombinant=20
HA proteins bind to human tracheal airway =
epithelium.</SPAN></STRONG></P>
<P =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/fig[2]/caption[1]/p[1]=
">(A)=20
Morphological features of human tracheal airway epithelium ex vivo =
visualized by=20
H&amp;E counterstain indicating cell-types present: (C) ciliated cell, =
(NC)=20
non-ciliated cell, (MS) mucin secreting cell. Histological sections of =
<I>ex=20
vivo</I> human tracheal tissue were probed with recombinant HA proteins =
cloned=20
from human H3 (B), or avian H5 (C) influenza viruses or with H5 HA =
receptor=20
binding mutants (D=E2=80=93G). Ciliated cells are identified with anti =
acetylated=20
=CE=B1-tubulin (green) and the HA-Fc proteins detected with anti =
human-Fc (red).=20
White arrows indicate cells that exhibit HA binding. Open arrowheads =
indicate=20
ciliated cells with HA binding, solid arrowheads indicate non-ciliated =
cells=20
with HA binding and the small arrow indicates binding to tops of cilia. =
These=20
are representative images and similar patterns were seen in repeated=20
experiments. (H) Numbers and types of epithelial cells bound by HA =
proteins were=20
quantified by counting 100=E2=80=93200 total cells from five different =
fields of=20
view.</P><SPAN=20
xpathLocation=3D"noSelect">doi:10.1371/journal.pone.0007836.g004</SPAN>
<DIV class=3Dclearer></DIV></DIV>
<P xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/p[2]">Since the =
recombinant=20
HA-Fc proteins should represent more biologically relevant ligands than =
lectins=20
for airway surface SAs utilized by influenza viruses, we used the HA-Fc =
fusion=20
proteins to probe the apical surfaces of HAE, detecting HA binding with =
an=20
anti-human IgG antibody conjugate. The dimeric HAs were pre-complexed =
into=20
larger multimers by mixing with anti-human Fc antibodies before =
application to=20
the tissue. When applied <I>en face</I>, H3 HA-Fc predominately bound to =

non-ciliated cells (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g003">Figure=20
3B</A>), a finding that was confirmed when histological sections of HAE =
were=20
similarly probed and immuno-reactivity for H3 HA-Fc localised =
predominately to=20
the apical surface of non-ciliated cells (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g003">Figure=20
3C</A>). In contrast, avian H5 HA-Fc bound sporadically to the apical =
surfaces=20
of HAE and was only detected in association with cells that co-stained =
with=20
acetylated =CE=B1-tubulin, i.e., ciliated cells (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g003">Figure=20
3D and 3E</A>). Binding of H5 HA was concentrated at the levels of the=20
microvilli on ciliated cells as opposed to the shafts of cilia (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g003">Figure=20
3E</A>), similar to the binding of the MAA lectin previously described =
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Thompson1">[15]</A>.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/p[3]">H5 HA =
mutants Q226L=20
and S227N bound to ciliated cells at the level of the microvilli similar =
to the=20
parental avian H5 HA (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g003">Figure=20
3F and 3G</A>). However, the G228S mutant showed an expanded binding =
profile=20
that included binding to non-ciliated cell types in addition to ciliated =
cells=20
(<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g003">Figure=20
3H</A>). The HA mutant bearing G228S in combination with the Q226L =
mutation also=20
bound to both ciliated and non-ciliated cells (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g003">Figure=20
3I and 3J</A>) indicating a partial switch in cell tropism.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[2]/p[4]">The binding =
of each=20
mutant to the different cell types was quantified by counting at least =
five=20
different fields of view containing a total of 100=E2=80=93200 cells. <A =

href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g003">Figure=20
3K</A> shows that the H3 HA protein bound to approximately 60% of the =
HAE cells=20
and the majority of those bound were of the non-ciliated type. In =
contrast all=20
of the H5 HA proteins bound to fewer cells and of the cells bound the =
majority=20
were ciliated although the mutants containing the G228S change bound to =
some=20
cells that were non-ciliated (dark blue).</P>
<H4 xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[3]/title[1]">Binding =
of HA=20
proteins to human airway epithelium tissue sections</H4>
<P xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[3]/p[1]">To determine =
if HA=20
interaction with HAE cultures is predictive for human airway epithelium =
<I>in=20
vivo</I> we next probed histological sections of freshly excised human =
tracheal=20
airway tissue with recombinant HA-Fc. Similar cell types were present in =
the=20
tracheal airway tissues as observed in HAE cultures confirming published =
data=20
that HAE cultures recapitulate the morphology of the tracheal airway =
epithelium=20
(<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g004">Figure=20
4A</A>). When HA-Fc were used to probe the human tracheal airway =
sections, H3=20
HA-Fc bound extensively to the luminal surface of the tissue and was =
localized=20
at the apical surfaces of cells. H3 HA bound predominantly to =
non-ciliated cells=20
in the tracheal sections, but also bound to some ciliated cells both at =
the=20
level of microvilli as well as, in some cases, to the tips of the cilia =
(<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g004">Figure=20
4B</A>). In contrast, H5 HA-Fc bound only sporadically to ciliated cells =
at the=20
level of microvilli (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g004">Figure=20
4C</A>). The single mutants Q226L and S227N bound to ciliated cell =
microvillus=20
regions (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g004">Figure=20
4D and 4E</A>), similar to wild-type H5 HA (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g003">Figure=20
3E</A>). In addition, some binding of the S227N mutant to cilia was =
observed (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g004">Figure=20
4E</A>). In contrast, HA with the G228S single mutation bound to both =
ciliated=20
cells and non-ciliated cells (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g004">Figure=20
4G</A>). The combined Q226L/G228S mutation exhibited a strikingly =
similar=20
binding pattern to that shown by the H3 HA, with binding predominately =
localized=20
to the apical surface of non-ciliated cells (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g004">Figure=20
4G versus 4B</A>).</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[3]/p[2]">Once again =
the extent=20
of binding of each mutant to the different cell types was quantified by =
counting=20
cells in at least five different fields of view (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g004">Figure=20
4H</A>). This clearly demonstrated that the double mutant Q226L/G228S =
bound both=20
to more cells in the section and showed an altered preference for the=20
non-ciliated cell type in comparison with the wild type H5 HA. The =
extent of=20
binding by G228S and the double mutant approached but did not reach the =
55% of=20
cells bound by the human adapted H3HA. Moreover although the S227N HA =
mutant=20
showed some binding to the cilia tips (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g004">Figure=20
4E</A>), analysis of several fields of cells obtained from different =
patients=20
did not reveal any binding to the non-ciliated cell type, in contrast to =
the=20
G228S containing mutants.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[3]/p[3]">To confirm =
that the=20
binding of HA-Fc observed in these studies was SA dependent, tracheal =
airway=20
sections were exposed to recombinant neuraminidase before probing with =
HA-Fc.=20
Treatment with neuraminidase (NA) cloned from <I>Salmonella =
typhimurium</I> LT2,=20
which displays a 260-fold substrate preference for =CE=B12-3 SA over =
=CE=B12-6 SA <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Hoyer1">[35]</A>=20
abrogated binding of H5 HA-Fc to human trachea epithelium (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g005">Figure=20
5B, middle panel</A>). Binding of H3 HA-Fc to the tops of cilia was also =

markedly reduced after treatment with the =CE=B12-3 specific NA, but =
binding to the=20
apical surface of cells lacking cilia persisted (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g005">Figure=20
5A, middle panel</A>). However, treatment with a broad specificity NA =
isolated=20
from <I>Clostridium perfringens</I> abolished both the H3 and H5 HA =
binding (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g005">Figure=20
5A and 5B, right hand panel</A>). The binding of the single or double =
mutants=20
containing the G228S mutation on ciliated cells was abolished by =
treatment with=20
=CE=B12-3-specific NA (compare <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g005">Figure=20
5C and 5D, left panel</A>, open triangles, with <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g005">Figure=20
5C and 5D, middle panel</A>). However, binding of these HA proteins to=20
non-ciliated cells (solid triangles) remained unless the broad =
specificity NA=20
from <I>C. perfringens</I> was used (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g005">Figure=20
5C and 5D, right panel</A>).</P>
<DIV class=3Dfigure =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[3]/fig[1]"><A=20
id=3Dpone-0007836-g005 title=3D"Click for larger image "=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g005"=20
name=3Dpone-0007836-g005><IMG class=3Dthumbnail border=3D1 =
alt=3Dthumbnail align=3Dleft=20
src=3D"http://www.plosone.org/article/fetchObject.action?uri=3Dinfo:doi/1=
0.1371/journal.pone.0007836.g005&amp;representation=3DPNG_S"=20
xpathLocation=3D"noSelect"></A>
<P><STRONG =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[3]/fig[1]/label[1]"><A=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g005"><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[3]/fig[1]/label[1]">Figur=
e 5.=20
</SPAN></A><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[3]/fig[1]/caption[1]/titl=
e[1]">Recombinant=20
HA proteins from human and avian influenza viruses bind <I>ex vivo</I> =
human=20
tracheal epithelium in a sialic acid dependant =
manner.</SPAN></STRONG></P>
<P=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[3]/fig[1]/caption[1]/p[1]=
">Histological=20
sections of <I>ex vivo</I> human tracheal epithelium were probed with =
human H3=20
(A), avian H5 (B) or H5 mutants, (C) G228S, and (D) Q226L/G228S, with =
and=20
without prior neuraminidase (NA) treatment. Slides were pre-incubated =
for 3=20
hours with 5 units of recombinant NA cloned from <I>Salmonella =
typhimurium</I>=20
LT2 (NEBL) that shows a 260-fold preference for =CE=B12-3 over =CE=B12-6 =
linked SA or with=20
15 units of recombinant NA cloned from <I>Clostridium perfringens</I> =
(NEBL)=20
that cleaves both =CE=B12-3 and =CE=B12-6 SA linkages. HA was detected =
as before using=20
anti-human Fc (red) and anti-acetylated =CE=B1-tubulin was used to =
indicate ciliated=20
cells (green). White arrows indicate cells that exhibit HA binding. Open =

arrowheads indicate ciliated cells with HA binding; solid arrowheads =
indicate=20
non-ciliated cells with HA binding.</P><SPAN=20
xpathLocation=3D"noSelect">doi:10.1371/journal.pone.0007836.g005</SPAN>
<DIV class=3Dclearer></DIV></DIV>
<H4 =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[4]/title[1]">Analysis of =
H5 HA=20
codon usage at amino acid 228</H4>
<P xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[4]/p[1]">Our data =
show a=20
specific amino acid mutation in H5 HA is sufficient to switch an avian=20
virus-like binding profile (ciliated cells) to a human virus-like =
binding=20
profile (ciliated and non-ciliated cells) in tracheal airway epithelium =
<I>in=20
vitro</I> and <I>ex vivo</I>. The mutation enabling this event is a =
change in=20
amino acid position 228 from G to S which likely increases the affinity =
of HA=20
for the long chain =CE=B12-6 SA lactosaminoglycans in the previously =
reported=20
=E2=80=98umbrella=E2=80=99 topology <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Stevens2">[21]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Chandrasekaran1">[24]</A>.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[4]/p[2]">While change =
of H5 HA=20
specificity is experimentally possible, the propensity for the requisite =

mutations to occur <I>in vivo</I> depends on the feasibility of the =
circulating=20
strains of H5N1 virus acquiring the necessary nucleotide changes. We =
assessed=20
whether principles of codon usage would enable such a mutation to occur =
in the=20
context of natural virus. An amino acid switch from glycine to serine at =
residue=20
228 requires either one or two nucleotide changes depending on the =
nature of the=20
third =E2=80=98wobble=E2=80=99 nucleotide in the glycine codon. All of =
the 1374 influenza H5 HA=20
gene sequences deposited in the available databases show that H5 viruses =
encode=20
G228 using codons GGA (93%) or GGG (7%) (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g006">Figure=20
6A</A>). Therefore, generation of a G228S mutation in H5 HA must involve =
one of=20
three possibilities: 1) Simultaneous mutation of two nucleotides, the =
frequency=20
of which is calculated to be lower than the maximum viral load of =
10<SUP>8</SUP>=20
copies of vRNA would allow <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-deJong1">[3]</A>,=20
assuming an error rate of 10<SUP>=E2=88=925</SUP> mutations per site <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Parvin1">[36]</A>;=20
2) Transversion at the third nucleotide of the codon to generate a =
neutral=20
intermediate that is unlikely to become enriched within the =
quasispecies; or, 3)=20
Passage through an intermediate in which either the first nucleotide of =
the=20
codon mutates initially to encode arginine by transition, or the second=20
nucleotide of the codon mutates by transversion to encode alanine (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g006">Figure=20
6A</A>). To assess the latter possibilities, we generated H5 HA-Fcs =
representing=20
the G228R and G228A intermediates and tested their binding to synthetic=20
glycopolymers, HAE and human tracheal airway tissue (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g006">Figure=20
6B=E2=80=936G</A>). Mutant G228A was unaffected in synthetic ligand =
binding compared to=20
wild-type H5 HA (<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g006">Figure=20
6E</A> versus <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g002">Figure=20
2</A>) and bound sporadically to ciliated cells in HAE and tracheal =
tissue.=20
Thus, this amino acid change would confer no advantage to a virus in the =
human=20
respiratory tract. Mutant G228R failed to bind any of the synthetic =
ligands and=20
did not bind any cells in HAE or tracheal tissue. Moreover, when the =
mutated H5=20
HA genes were used in attempts to rescue live influenza virus using =
reverse=20
genetics we were unable to rescue viruses containing the G228R mutation. =
This is=20
consistent with the notion that a G228R H5 HA is not capable of =
generating a=20
viable intermediate during the evolution of a receptor switched virus. =
In=20
contrast, viruses with wild-type H5 HA RBS or bearing other amino acid =
changes=20
at residue 228 such as G228A or G228S were rescued (data not shown).</P>
<DIV class=3Dfigure =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[4]/fig[1]"><A=20
id=3Dpone-0007836-g006 title=3D"Click for larger image "=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g006"=20
name=3Dpone-0007836-g006><IMG class=3Dthumbnail border=3D1 =
alt=3Dthumbnail align=3Dleft=20
src=3D"http://www.plosone.org/article/fetchObject.action?uri=3Dinfo:doi/1=
0.1371/journal.pone.0007836.g006&amp;representation=3DPNG_S"=20
xpathLocation=3D"noSelect"></A>
<P><STRONG =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[4]/fig[1]/label[1]"><A=20
onclick=3D"window.open(this.href,'plosSlideshow','directories=3Dno,locati=
on=3Dno,menubar=3Dno,resizable=3Dyes,status=3Dno,scrollbars=3Dyes,toolbar=
=3Dno,height=3D600,width=3D850');return false;"=20
href=3D"http://www.plosone.org/article/slideshow.action?uri=3Dinfo:doi/10=
.1371/journal.pone.0007836&amp;imageURI=3Dinfo:doi/10.1371/journal.pone.0=
007836.g006"><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[4]/fig[1]/label[1]">Figur=
e 6.=20
</SPAN></A><SPAN=20
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[4]/fig[1]/caption[1]/titl=
e[1]">Nucleotide=20
sequence changes required to switch H5 HA receptor binding=20
preference.</SPAN></STRONG></P>
<P =
xpathLocation=3D"/article[1]/body[1]/sec[2]/sec[4]/fig[1]/caption[1]/p[1]=
">(A)=20
Analysis of codon usage by 1374 H5 influenza viruses indicates that =
codon 228 is=20
GGA in 93% strains and GGG in 7%. The table illustrates transversions =
and=20
transitions by which coding capacity at this residue might change from =
glycine=20
to serine. The ability of the two intermediate mutants 228R (B) and 228A =
(E)=20
recombinant HA proteins to bind synthetic receptor ligands 3SLN (avian =
receptor)=20
and 6SLN (human receptor) in a solid phase assay was assessed. The =
binding=20
phenotype on HAE (C,F) or human tracheal epithelium (D,G) by recombinant =

proteins with each amino acid sequence changed at residue 228 are shown =
(C,D)=20
G228R and (F,G) G228A.</P><SPAN=20
xpathLocation=3D"noSelect">doi:10.1371/journal.pone.0007836.g006</SPAN>
<DIV class=3Dclearer></DIV></DIV></DIV>
<DIV id=3Dsection3 xpathLocation=3D"/article[1]/body[1]/sec[3]"=20
xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema"><A id=3Ds3 =
title=3DDiscussion name=3Ds3=20
toc=3D"s3"></A>
<H3 xpathLocation=3D"noSelect">Discussion&nbsp;<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#top">Top</A></H3>
<P xpathLocation=3D"/article[1]/body[1]/sec[3]/p[1]">We address =
potential reasons=20
why H5N1 avian influenza virus has not yet adapted for human-to-human=20
transmission in spite of the unprecedented distribution and frequent =
zoonoses of=20
this avian influenza virus. Text book explanations of the origins of =
influenza=20
virus pandemics describe that reassortment of genes from avian strains =
and human=20
strains give rise to pandemic viruses. One possible explanation why this =
event=20
has not occurred for H5N1 could be an incompatibility between the H5 HA =
gene and=20
other genes of currently circulating human influenza viruses. However,=20
laboratory-generated recombinant viruses have shown that surface antigen =
genes=20
from South East Asian H5N1 viruses are compatible with internal viral =
genes from=20
currently circulating H3N2 human strains and coinfection of ferret in a=20
laboratory setting produced <I>in vivo</I> reassortants with mixtures of =
genes=20
derived from human circulating virus and H5N1 <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Chen1">[37]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Jackson1">[38]</A>.=20
Whether there is an impediment to the reassortment happening in nature =
is not=20
resolved. In addition, an increasing body of work supports the notion =
that=20
reassortment <I>per se</I> is not sufficient to generate a pandemic.=20
Importantly, both unmodified H5N1 avian influenza viruses and laboratory =

generated chimeric human: avian reassortant viruses with unmodified H5N1 =
surface=20
antigens failed to transmit between ferrets, the most relevant animal =
model for=20
studies of human-like transmissibility of influenza viruses <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Yen1">[39]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Maines1">[40]</A>.=20
Moreover, similar work with other avian subtypes of influenza virus =
indicate=20
that mutations in the receptor binding site of HA that switch the SA =
preference=20
are required for efficient transmission in the ferret model. For =
example, avian=20
influenza viruses of the H7 subtype North American lineage that have =
undergone=20
such a receptor switch showed transmission between ferrets in a direct =
contact=20
model <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Belser1">[41]</A>.=20
Similarly, H9N2 viruses whose HA genes display enhanced preference for =
=CE=B12-6SA=20
binding were transmitted between co-housed ferrets whereas viruses with =
HAs that=20
bound to =CE=B12-3SA were not <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Wan1">[12]</A>.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[3]/p[2]">By analogy with =
these other=20
avian subtypes and in light of the experiences of previous pandemics <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Matrosovich1">[8]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Tumpey1">[9]</A>=20
we judge that adaptation of H5 HA at the RBS will be required for =
human-to-human=20
transmissibility. Our data, summarized in <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone-0007836-g006">Figure=20
6</A>, provide a rational explanation of why H5 HA has not yet adapted =
in this=20
way. By analysis of codon usage required to generate mutants capable of =
SA=20
switching, we found that two or more simultaneous nucleotide changes are =

required to adapt the H5 HA gene for efficient binding to cells of the =
human=20
tracheal epithelium. Moreover, our data indicate that single nucleotide =
change=20
intermediates would be predicted to reduce viral fitness in the =
environment of=20
the human airways. Indeed, attempts to generate such single nucleotide =
mutants=20
in the context of infectious H5N1 virus indicate they are either =
non-viable or=20
display attenuated growth kinetics (unpublished work).</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[3]/p[3]">Although some =
naturally=20
occurring H5 HA mutants with increased affinity for synthetic human =
receptor=20
mimics, including the S227N mutation considered here, have been detected =
within=20
the quasispecies of viruses isolated from H5N1 infected patients <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Gambaryan1">[33]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Auewarakul1">[42]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Yamada1">[43]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Kongchanagul1">[44]</A>,=20
none of them have resulted in documented human-to-human transmission of =
the=20
virus suggesting that acquisition of binding to synthetic ligands is not =
an=20
adequate predictor of transmissibility. Mutations spread throughout the =
HA=20
protein have been found to affect receptor binding affinity and =
preference in=20
the context of other HA subtypes <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Matrosovich1">[8]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Tumpey1">[9]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Ibricevic1">[13]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Medeiros1">[14]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Thompson1">[15]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Stevens1">[20]</A>.=20
Although we cannot claim that the mutations we describe are the only =
ones that=20
will switch receptor binding preference of H5 HA, extensive =
investigation of=20
pandemic evolution <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Matrosovich4">[45]</A>=20
coupled with the cumulative research of several groups <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Harvey1">[18]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Stevens1">[20]</A>=20
suggest they are the most likely candidates. Moreover, the failure of =
H5N1 to=20
emerge as a pandemic strain despite the wide variety of species infected =
since=20
2003 suggests that there is a significant natural barrier to the =
evolution of H5=20
HA receptor binding specificity.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[3]/p[4]">The reasons why HA =
of avian=20
influenza virus has to adapt its RBS before acquiring human =
transmissibility are=20
still not explained in detail. Studies utilizing lectin probes suggest a =
paucity=20
of =CE=B12-3SA in the upper respiratory tract <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Tumpey1">[9]</A>.=20
Our data shows that H5 HA protein can bind to =CE=B12-3SA on human =
airway epithelium=20
but that the cell specificity of the binding differs from that shown by =
a human=20
influenza HA. Interestingly the binding of H5 HA for synthetic =
=CE=B12-3sialylated=20
glycans showed a different pattern than that shown for the human =
para-influenza=20
viruses that also bind to the ciliated cell in human airway and transmit =

effectively between humans <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Zhang1">[27]</A>=20
Although replication of H5N1 virus in human nasal tissue <I>ex vivo</I> =
has been=20
demonstrated <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Nicholls1">[23]</A>,=20
the scenario during actual transmission events between human hosts may =
be more=20
limiting. In these cases, virus attachment might only occur to certain =
cell=20
types that are in a specific anatomical location or that most abundantly =
display=20
the relevant ligand.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[3]/p[5]">Our findings that =
mutations=20
in the H5 HA gene can alter receptor binding in the human airway =
epithelium may=20
have implications for the generation of effective live attenuated =
vaccines.=20
Although they are unlikely to be used during the pre-pandemic period due =
to the=20
risk that reassortment with circulating strains might generate a novel =
virus=20
with pandemic potential itself, live attenuated H5N1 vaccines may =
represent an=20
important dose sparing strategy for wide scale vaccination of the =
population in=20
the face of an ongoing pandemic with highly pathogenic H5N1 virus. =
Indeed, cold=20
adapted influenza viruses with wild-type H5N1 surface antigens have been =
tested,=20
but their immunogenicity in ferrets and in humans have been =
disappointing to=20
date <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Lee1">[46]</A>.=20
Using reverse genetics, the Q226L and G228S mutations could be =
artificially=20
engineered in recombinant H5 HA within the cold adapted backbone. This =
may=20
increase the vaccine virus' ability to replicate in the human upper =
respiratory=20
tract and thus induce a more substantive immune response leading to the=20
effective immunological memory required for good vaccine efficacy.</P>
<P xpathLocation=3D"/article[1]/body[1]/sec[3]/p[6]">H5N1 continues to =
circulate=20
and diversify <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-WHO2">[2]</A>=20
and some mutations accrued since 2004 may enhance the virus' capacity =
for=20
receptor switching <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Stevens2">[21]</A>.=20
It is also noteworthy that lesser studied subtypes of avian influenza =
may be=20
evolutionarily closer to receptor switching than previously assumed <A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Bateman1">[11]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Wan1">[12]</A>,=20
<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#pone.0007836-Belser1">[41]</A>.=20
Tissue binding assays such as those described here will be useful in =
assessing=20
the pandemic potential of such viruses. The widespread preparedness for =
an=20
influenza pandemic that has resulted from the prevalence of H5N1 =
therefore=20
remains appropriate even in light of the data we present.</P></DIV>
<DIV id=3Dsection4 xpathLocation=3D"/article[1]/body[1]/sec[4]"=20
xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema"><A id=3Ds4=20
title=3D"Materials and Methods" name=3Ds4 toc=3D"s4"></A>
<H3 xpathLocation=3D"noSelect">Materials and Methods&nbsp;<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#top">Top</A></H3>
<H4 =
xpathLocation=3D"/article[1]/body[1]/sec[4]/sec[1]/title[1]">Expression =
of=20
influenza HA protein in insect cells</H4>
<P =
xpathLocation=3D"/article[1]/body[1]/sec[4]/sec[1]/p[1]"><I>Spodoptera=20
frugiperda</I> cells (Sf9) were grown in suspension in BioWhittaker=20
Insect-Xpress serum-free media (Cambrex) supplemented with 2% fetal calf =
serum.=20
They were infected with recombinant baculovirus expressing H3-Fc, H5-Fc =
or H5-Fc=20
receptor binding mutants and incubated at 28=C2=B0C. After 72 h the =
supernatant was=20
harvested by centrifugation at 4000 rpm for 10 min to remove the =
infected cells.=20
The soluble protein was concentrated by centrifugation at 4500 rpm for 4 =
h using=20
Vivaspin columns (Viva Science, Sartorius Group). The yield of =
concentrated=20
protein ranged from 500=E2=80=93700 =C2=B5g/mL. Protein was aliquoted =
and stored at=20
=E2=88=9280=C2=B0C.</P>
<H4 xpathLocation=3D"/article[1]/body[1]/sec[4]/sec[2]/title[1]">Binding =
of HA-Fc=20
proteins to human airway epithelial culture and tracheal airway tissue=20
sections</H4>
<P xpathLocation=3D"/article[1]/body[1]/sec[4]/sec[2]/p[1]">Paraffin =
embedded=20
sections of cultured human airway epithelium or normal human tracheal =
sections=20
were obtained from UNC Cystic Fibrosis Center Core facility. HAE derived =
from at=20
least 3 codes and tracheal sections from 3 individuals were used in this =
study=20
and representative images are shown. There was no notable difference =
between=20
cultures or sections from different individuals. The sections were=20
deparaffinized and rehydrated. Non-specific binding was blocked using 3% =
bovine=20
serum albumin (BSA) in phosphate-buffered saline (PBS) for 1 hour at =
room=20
temperature (RT) before incubation with ligands. Concentrated =
baculovirus=20
supernatants containing H3, H5 or H5 HA and mutant proteins were =
pre-complexed=20
with goat anti-human IgG (Fc specific) antibody (Invitrogen) at a ratio =
of 5 =C2=B5g=20
of HA-Fc: 0.6 =C2=B5g of antibody, for 1 hour at 4=C2=B0C. Sections were =
incubated with=20
pre-complexed HA-Fc in 1% BSA in PBS ON at 4=C2=B0C or RT for 3 hours. =
After washing=20
in PBS-T, slides were incubated with rabbit anti-acetylated =
=CE=B1-tubulin in 1% BSA=20
in PBS for 3 hours RT or overnight (ON) at 4=C2=B0C. Following washing =
with PBS-T=20
(PBS with 0.05% Tween-20), antibody-incubated slides were immersed in 1% =
BSA in=20
PBS containing donkey anti-goat FITC (Immunologicals Direct) and donkey=20
anti-rabbit Alexafluor 647 (Invitrogen) for 1 hour at RT. Slides were =
washed=20
with PBS-T and viewed under a confocal microscope. Images were collected =
on a=20
Zeiss Pascal LSM5 laser scanning microscope using Axioplan2 imaging =
software=20
using a Plan-Apochromat 63=C3=971.4 oil Ph3 lens. The binding of each =
mutant to the=20
different cell types was quantified by counting at least five different =
fields=20
of view containing a total of 100=E2=80=93200 cells.</P>
<H4 =
xpathLocation=3D"/article[1]/body[1]/sec[4]/sec[3]/title[1]">Sialidase=20
treatment of tissue sections</H4>
<P xpathLocation=3D"/article[1]/body[1]/sec[4]/sec[3]/p[1]">Five units =
of =CE=B12-3=20
sialic acid-specific neuraminidase from <I>Salmonella typhimurium</I> or =
15=20
units of a =CE=B12-3/=CE=B12-6 dual specificity enzyme from =
<I>Clostridium perfingens</I>=20
were incubated with deparaffinized sections for 3 hours at 37=C2=B0C. =
Sections were=20
then washed in PBS-T before being probed with recombinant HA proteins as =

above.</P></DIV>
<DIV xpathLocation=3D"noSelect" =
xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema"><A id=3Dack =
title=3DAcknowledgments=20
name=3Dack toc=3D"ack"></A>
<H3 xpathLocation=3D"noSelect">Acknowledgments&nbsp;<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#top">Top</A></H3>
<P xpathLocation=3D"/article[1]/back[1]/ack[1]/p[1]">All confocal work =
was=20
undertaken in the Henry Wellcome Imaging centre at St Marys, Imperial =
College=20
London. We thank Michael Hollinshead for technical assistance in the =
Imaging=20
Centre. We are grateful to the Directors and teams of the UNC Cystic =
Fibrosis=20
Center Core Facilities (Tissue Culture Core, Morphology and Morphometry =
Core and=20
the Michael Hooker Microscopy Facility) for supplying reagents and =
technical=20
expertise.</P></DIV>
<DIV class=3Dcontributions xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema"><A id=3Dauthcontrib=20
title=3D"Author Contributions" name=3Dauthcontrib =
toc=3D"authcontrib"></A>
<H3 xpathLocation=3D"noSelect">Author Contributions&nbsp;<A=20
href=3D"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007=
836#top">Top</A></H3>
<P xpathLocation=3D"noSelect"><SPAN class=3Dcapture-id>Conceived and =
designed the=20
experiments: GAT HS MAS IMJ RJP WSB. Performed the experiments: GAT HS =
MAS.=20
Analyzed the data: GAT HS MAS RJP WSB. Contributed =
reagents/materials/analysis=20
tools: MAS JR IMJ RJP. Wrote the paper: GAT HS MAS IMJ RJP =
WSB.</SPAN></P></DIV>
<DIV xpathLocation=3D"noSelect" =
xmlns:aml=3D"http://topazproject.org/aml/"=20
xmlns:mml=3D"http://www.w3.org/1998/Math/MathML"=20
xmlns:xlink=3D"http://www.w3.org/1999/xlink"=20
xmlns:xs=3D"http://www.w3.org/2001/XMLSchema"><A id=3Dreferences =
title=3DReferences=20
name=3Dreferences toc=3D"references"></A>
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}
.dijitSpinner .dijitInputLayoutContainer .dijitDownArrowButton {
	BOTTOM: 0px; BORDER-TOP-WIDTH: 1px !important
}
.dijitSpinner .dijitInputLayoutContainer .dijitUpArrowButton {
	TOP: 0px
}
.dijitComboButton TABLE {
	PADDING-BOTTOM: 0px; BORDER-RIGHT-WIDTH: 0px; MARGIN: 0px; =
PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BORDER-COLLAPSE: separate; =
BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; =
PADDING-TOP: 0px
}
.dijitComboButton .dijitButtonContents {
	BORDER-RIGHT-WIDTH: 0px !important
}
.dj_ie BUTTON.dijitButtonNode {
	OVERFLOW: visible
}
TABLE .dijitButton .dijitButtonNode {
	OVERFLOW: hidden
}
TABLE .dijitComboButton .dijitButtonNode {
	OVERFLOW: hidden
}
.dijitButtonNode IMG {
	MARGIN-BOTTOM: 0.2em; VERTICAL-ALIGN: middle
}
.dijitTextBox {
	BORDER-BOTTOM: black 1px solid; BORDER-LEFT: black 1px solid; WIDTH: =
15em; OVERFLOW: hidden; BORDER-TOP: black 1px solid; BORDER-RIGHT: black =
1px solid
}
.dijitComboBox {
	BORDER-BOTTOM: black 1px solid; BORDER-LEFT: black 1px solid; WIDTH: =
15em; OVERFLOW: hidden; BORDER-TOP: black 1px solid; BORDER-RIGHT: black =
1px solid
}
.dijitSpinner {
	BORDER-BOTTOM: black 1px solid; BORDER-LEFT: black 1px solid; WIDTH: =
15em; OVERFLOW: hidden; BORDER-TOP: black 1px solid; BORDER-RIGHT: black =
1px solid
}
.dijitTextBox INPUT:focus {
	OUTLINE-STYLE: none
}
.dijitComboBox INPUT:focus {
	OUTLINE-STYLE: none
}
.dijitSpinner INPUT:focus {
	OUTLINE-STYLE: none
}
.dijitTextBoxFocused {
=09
}
.dijitComboBoxFocused {
=09
}
.dijitSpinnerFocused {
=09
}
.dijitTextBox INPUT {
	BACKGROUND-IMAGE: none !important; POSITION: static !important; =
BORDER-LEFT: black 0px solid; PADDING-BOTTOM: 0px; BORDER-RIGHT-WIDTH: =
0px !important; BACKGROUND-COLOR: transparent !important; MARGIN: 0px; =
PADDING-LEFT: 0px; WIDTH: 100% !important; PADDING-RIGHT: 0px; DISPLAY: =
inline; BORDER-TOP-WIDTH: 0px !important; BORDER-BOTTOM-WIDTH: 0px =
!important; VISIBILITY: visible !important; VERTICAL-ALIGN: 0em =
!important; PADDING-TOP: 0px
}
.dijitComboBox INPUT {
	BACKGROUND-IMAGE: none !important; POSITION: static !important; =
BORDER-LEFT: black 0px solid; PADDING-BOTTOM: 0px; BORDER-RIGHT-WIDTH: =
0px !important; BACKGROUND-COLOR: transparent !important; MARGIN: 0px; =
PADDING-LEFT: 0px; WIDTH: 100% !important; PADDING-RIGHT: 0px; DISPLAY: =
inline; BORDER-TOP-WIDTH: 0px !important; BORDER-BOTTOM-WIDTH: 0px =
!important; VISIBILITY: visible !important; VERTICAL-ALIGN: 0em =
!important; PADDING-TOP: 0px
}
.dijitSpinner INPUT {
	BACKGROUND-IMAGE: none !important; POSITION: static !important; =
BORDER-LEFT: black 0px solid; PADDING-BOTTOM: 0px; BORDER-RIGHT-WIDTH: =
0px !important; BACKGROUND-COLOR: transparent !important; MARGIN: 0px; =
PADDING-LEFT: 0px; WIDTH: 100% !important; PADDING-RIGHT: 0px; DISPLAY: =
inline; BORDER-TOP-WIDTH: 0px !important; BORDER-BOTTOM-WIDTH: 0px =
!important; VISIBILITY: visible !important; VERTICAL-ALIGN: 0em =
!important; PADDING-TOP: 0px
}
.dijitValidationIcon {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 2px; PADDING-RIGHT: 2px; DISPLAY: =
block; FLOAT: right; HEIGHT: auto; VISIBILITY: hidden; PADDING-TOP: 0px
}
.dijitValidationIconText {
	LINE-HEIGHT: 190%; FONT-STYLE: italic; DISPLAY: none; FONT-FAMILY: =
sans-serif; FLOAT: right; VISIBILITY: hidden; FONT-SIZE: 0.45em
}
.dijit_a11y .dijitValidationIcon {
	DISPLAY: none !important
}
.dijit_a11y .dijitValidationIconText {
	DISPLAY: block !important
}
.dijitError .dijitValidationIcon {
	VISIBILITY: visible
}
.dijitError .dijitValidationIconText {
	VISIBILITY: visible
}
.dijitTextBox .dijitArrowButton {
	DISPLAY: none
}
.dijitCheckBox {
	PADDING-BOTTOM: 0px; BORDER-RIGHT-WIDTH: 0px; PADDING-LEFT: 0px; WIDTH: =
16px; PADDING-RIGHT: 0px; BACKGROUND-REPEAT: no-repeat; =
BACKGROUND-POSITION: center center; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; HEIGHT: 16px; OVERFLOW: hidden; =
BORDER-LEFT-WIDTH: 0px; PADDING-TOP: 0px
}
.dijitRadio {
	PADDING-BOTTOM: 0px; BORDER-RIGHT-WIDTH: 0px; PADDING-LEFT: 0px; WIDTH: =
16px; PADDING-RIGHT: 0px; BACKGROUND-REPEAT: no-repeat; =
BACKGROUND-POSITION: center center; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; HEIGHT: 16px; OVERFLOW: hidden; =
BORDER-LEFT-WIDTH: 0px; PADDING-TOP: 0px
}
.dijitCheckBoxInput {
	PADDING-BOTTOM: 0px; BORDER-RIGHT-WIDTH: 0px; PADDING-LEFT: 0px; WIDTH: =
16px; PADDING-RIGHT: 0px; BACKGROUND-REPEAT: no-repeat; =
BACKGROUND-POSITION: center center; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; HEIGHT: 16px; OVERFLOW: hidden; =
BORDER-LEFT-WIDTH: 0px; PADDING-TOP: 0px
}
.dijitCheckBox INPUT {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; DISPLAY: block; PADDING-TOP: 0px
}
.dijitRadio INPUT {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; DISPLAY: block; PADDING-TOP: 0px
}
.dijitCheckBoxInput {
	opacity: 0.01
}
.dj_ie .dijitCheckBoxInput {
	FILTER: alpha(opacity=3D0)
}
.dijit_a11y .dijitCheckBox {
	WIDTH: auto; HEIGHT: auto
}
.dijit_a11y .dijitRadio {
	WIDTH: auto; HEIGHT: auto
}
.dijit_a11y .dijitCheckBoxInput {
	FILTER: none; WIDTH: auto; HEIGHT: auto; opacity: 1
}
.dijitProgressBarEmpty {
	Z-INDEX: 0; BORDER-BOTTOM: black 1px solid; POSITION: relative; =
BORDER-LEFT: black 1px solid; OVERFLOW: hidden; BORDER-TOP: black 1px =
solid; BORDER-RIGHT: black 1px solid
}
.dijitProgressBarFull {
	Z-INDEX: -1; POSITION: absolute; WIDTH: 100%; HEIGHT: 100%; OVERFLOW: =
hidden; TOP: 0px
}
.dijitProgressBarTile {
	POSITION: absolute; PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #aaa; =
MARGIN: 0px; PADDING-LEFT: 0px; WIDTH: auto; BOTTOM: 0px; =
BACKGROUND-ATTACHMENT: fixed; PADDING-RIGHT: 0px; HEIGHT: auto; =
OVERFLOW: hidden; TOP: 0px; RIGHT: 0px; PADDING-TOP: 0px; LEFT: 0px
}
.dijit_a11y .dijitProgressBarTile {
	BORDER-BOTTOM: 4px solid; BORDER-LEFT: 4px solid; BACKGROUND-COLOR: =
transparent !important; BORDER-TOP: 4px solid; BORDER-RIGHT: 4px solid
}
.dj_iequirks .dijitProgressBarTile {
	WIDTH: 100%; HEIGHT: 100%
}
.dj_ie6 .dijitProgressBarTile {
	POSITION: static; HEIGHT: 100%
}
.dijitProgressBarIndeterminate .dijitProgressBarLabel {
	VISIBILITY: hidden
}
.dijitProgressBarIndeterminate .dijitProgressBarTile {
=09
}
.dijitProgressBarIndeterminateHighContrastImage {
	DISPLAY: none
}
.dijit_a11y .dijitProgressBarIndeterminate =
.dijitProgressBarIndeterminateHighContrastImage {
	POSITION: absolute; PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: =
0px; WIDTH: 100%; BOTTOM: 0px; PADDING-RIGHT: 0px; DISPLAY: block; =
HEIGHT: auto; TOP: 0px; PADDING-TOP: 0px
}
.dijitProgressBarLabel {
	POSITION: static; TEXT-ALIGN: center; BACKGROUND-COLOR: transparent =
!important; WIDTH: 100%; DISPLAY: block
}
.dijitProgressBarV .dijitProgressBarFull {
	BOTTOM: 0px
}
.dj_ie6 .dijitProgressBarV .dijitProgressBarTile {
	POSITION: absolute; WIDTH: 100%
}
.dijitTooltip {
	Z-INDEX: 2000; POSITION: absolute; DISPLAY: block; OVERFLOW: visible; =
TOP: -10000px; LEFT: 50%
}
.dijitTooltipContainer {
	BORDER-BOTTOM: black 2px solid; BORDER-LEFT: black 2px solid; =
BACKGROUND: #b8b5b5; COLOR: black; FONT-SIZE: small; BORDER-TOP: black =
2px solid; BORDER-RIGHT: black 2px solid
}
.dijitTooltipFocusNode {
	PADDING-BOTTOM: 2px; PADDING-LEFT: 2px; PADDING-RIGHT: 2px; =
PADDING-TOP: 2px
}
.dijitTooltipConnector {
	POSITION: absolute
}
.dijitLayoutContainer {
	POSITION: relative; DISPLAY: block; OVERFLOW: hidden
}
BODY .dijitAlignTop {
	POSITION: absolute; OVERFLOW: hidden
}
BODY .dijitAlignBottom {
	POSITION: absolute; OVERFLOW: hidden
}
BODY .dijitAlignLeft {
	POSITION: absolute; OVERFLOW: hidden
}
BODY .dijitAlignRight {
	POSITION: absolute; OVERFLOW: hidden
}
BODY .dijitAlignClient {
	POSITION: absolute
}
.dijitBorderContainer {
	POSITION: relative; OVERFLOW: hidden
}
.dijitBorderContainer > .dijitTextArea {
	resize: none
}
.dijitSplitter {
	Z-INDEX: 10 !important; BORDER-BOTTOM: gray 0px solid; POSITION: =
absolute; BORDER-LEFT: gray 0px solid; BACKGROUND-COLOR: #fff; OVERFLOW: =
hidden; BORDER-TOP: gray 0px solid; BORDER-RIGHT: gray 0px solid
}
.dijitSplitterCover {
	Z-INDEX: 1; POSITION: absolute; WIDTH: 100%; HEIGHT: 100%; TOP: 0px; =
LEFT: 0px
}
.dijitSplitterH {
	BORDER-TOP-WIDTH: 1px; BORDER-BOTTOM-WIDTH: 1px; HEIGHT: 7px; CURSOR: =
ns-resize
}
.dijitSplitterV {
	BORDER-RIGHT-WIDTH: 1px; WIDTH: 7px; BORDER-LEFT-WIDTH: 1px; CURSOR: =
ew-resize
}
.dijitSplitContainer {
	POSITION: relative; DISPLAY: block; OVERFLOW: hidden
}
.dijitSplitPane {
	POSITION: absolute
}
.dijitSplitContainerSizerH {
	BORDER-BOTTOM: buttonshadow 1px solid; POSITION: absolute; BORDER-LEFT: =
buttonhighlight 1px solid; BACKGROUND-COLOR: buttonface; MARGIN: 0px; =
FONT-SIZE: 1px; BORDER-TOP: buttonhighlight 1px solid; CURSOR: w-resize; =
BORDER-RIGHT: buttonshadow 1px solid
}
.dijitSplitContainerSizerV {
	BORDER-BOTTOM: buttonshadow 1px solid; POSITION: absolute; BORDER-LEFT: =
buttonhighlight 1px solid; BACKGROUND-COLOR: buttonface; MARGIN: 0px; =
FONT-SIZE: 1px; BORDER-TOP: buttonhighlight 1px solid; CURSOR: w-resize; =
BORDER-RIGHT: buttonshadow 1px solid
}
.dijitSplitContainerSizerH .thumb {
	POSITION: absolute; OVERFLOW: hidden; TOP: 49%
}
.dijitSplitterV .dijitSplitterThumb {
	POSITION: absolute; OVERFLOW: hidden; TOP: 49%
}
.dijitSplitContainerSizerV .thumb {
	POSITION: absolute; LEFT: 49%
}
.dijitSplitterH .dijitSplitterThumb {
	POSITION: absolute; LEFT: 49%
}
.dijitSplitterShadow {
	FILTER: Alpha(Opacity=3D50); BACKGROUND-COLOR: buttonshadow; MARGIN: =
0px; FONT-SIZE: 1px; opacity: 0.5; -moz-opacity: 0.5
}
.dijitSplitContainerVirtualSizerH {
	FILTER: Alpha(Opacity=3D50); BACKGROUND-COLOR: buttonshadow; MARGIN: =
0px; FONT-SIZE: 1px; opacity: 0.5; -moz-opacity: 0.5
}
.dijitSplitContainerVirtualSizerV {
	FILTER: Alpha(Opacity=3D50); BACKGROUND-COLOR: buttonshadow; MARGIN: =
0px; FONT-SIZE: 1px; opacity: 0.5; -moz-opacity: 0.5
}
.dj_ie .dijitSplitterV {
	CURSOR: w-resize
}
.dijitSplitContainerVirtualSizerH {
	CURSOR: w-resize
}
.dj_ie .dijitSplitterH {
	CURSOR: n-resize
}
.dijitSplitContainerSizerV {
	CURSOR: n-resize
}
.dijitSplitContainerVirtualSizerV {
	CURSOR: n-resize
}
.dijit_a11y .dijitSplitterH {
	BORDER-BOTTOM: #d3d3d3 1px solid; BORDER-TOP: #d3d3d3 1px solid
}
.dijit_a11y .dijitSplitterV {
	BORDER-LEFT: #d3d3d3 1px solid; BORDER-RIGHT: #d3d3d3 1px solid
}
.dijitContentPane {
	DISPLAY: block; OVERFLOW: auto
}
.dijitTitlePane {
	DISPLAY: block; OVERFLOW: hidden
}
.dj_ie7 .dijitTitlePaneTextNode {
	DISPLAY: inline
}
.dijitColorPalette {
	BORDER-BOTTOM: #999 1px solid; BORDER-LEFT: #999 1px solid; BACKGROUND: =
#fff; BORDER-TOP: #999 1px solid; BORDER-RIGHT: #999 1px solid; =
-moz-border-radius: 3pt
}
IMG.dijitColorPaletteUnder {
	POSITION: absolute; BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: =
none; BORDER-TOP-STYLE: none; BORDER-LEFT-STYLE: none; TOP: 0px; LEFT: =
0px
}
.dijitColorPaletteInner {
	POSITION: relative; OUTLINE-WIDTH: 0px; OVERFLOW: hidden
}
.dijitPaletteCell {
	Z-INDEX: 10; POSITION: absolute; OUTLINE-WIDTH: 0px; WIDTH: 20px; =
HEIGHT: 20px; OVERFLOW: hidden
}
.dijitPaletteImg {
	BORDER-BOTTOM: #999 1px solid; POSITION: absolute; BORDER-LEFT: #999 =
1px solid; WIDTH: 16px; HEIGHT: 14px; OVERFLOW: hidden; BORDER-TOP: #999 =
1px solid; TOP: 1px; CURSOR: default; BORDER-RIGHT: #999 1px solid; =
LEFT: 1px
}
.dijitPaletteCellHighlight IMG {
	BORDER-BOTTOM: #000 2px solid; POSITION: absolute; BORDER-LEFT: #000 =
2px solid; OUTLINE-STYLE: solid; OUTLINE-COLOR: #dedede; OUTLINE-WIDTH: =
1px; WIDTH: 14px; HEIGHT: 12px; OVERFLOW: hidden; BORDER-TOP: #000 2px =
solid; TOP: 1px; CURSOR: default; BORDER-RIGHT: #000 2px solid; LEFT: =
1px
}
.dijit_a11y .dijitPaletteCell {
	BACKGROUND-COLOR: transparent !important
}
.dijit_a11y .dijitPaletteImg {
	BACKGROUND-COLOR: transparent !important
}
.dijitAccordionPane {
	OVERFLOW: hidden !important
}
.dijitAccordionPane .dijitAccordionBody {
	OVERFLOW: auto
}
.dijitAccordionContainer {
	BORDER-BOTTOM: #b7b7b7 1px solid; BORDER-LEFT: #b7b7b7 1px solid; =
BORDER-TOP: #b7b7b7 0px solid; BORDER-RIGHT: #b7b7b7 1px solid
}
.dijitAccordionTitle:hover {
	CURSOR: pointer
}
.dijitAccordionTitle .dijitAccordionArrow {
	FLOAT: right
}
.dijitAccordionPane .dijitAccordionTitle .arrowTextUp {
	DISPLAY: none; FLOAT: right; FONT-SIZE: 0.65em; FONT-WEIGHT: normal =
!important
}
.dijitAccordionPane .dijitAccordionTitle .arrowTextDown {
	DISPLAY: none; FLOAT: right; FONT-SIZE: 0.65em; FONT-WEIGHT: normal =
!important
}
.dijit_a11y .dijitAccordionPane .dijitAccordionTitle .arrowTextUp {
	DISPLAY: inline
}
.dijit_a11y .dijitAccordionTitle-selected .arrowTextDown {
	DISPLAY: inline
}
.dijit_a11y .dijitAccordionTitle-selected .arrowTextUp {
	DISPLAY: none
}
.dijitCalendarContainer THEAD TR TH {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
PADDING-TOP: 0px
}
.dijitCalendarContainer THEAD TR TD {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
PADDING-TOP: 0px
}
.dijitCalendarContainer TBODY TR TD {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
PADDING-TOP: 0px
}
.dijitCalendarContainer TFOOT TR TD {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
PADDING-TOP: 0px
}
.dijitCalendarNextYear {
	MARGIN: 0px 0px 0px 0.55em
}
.dijitCalendarPreviousYear {
	MARGIN: 0px 0.55em 0px 0px
}
.dijitCalendarIncrementControl {
	WIDTH: 1em; CURSOR: hand
}
.dijitCalendarDisabledDate {
	COLOR: gray !important
}
.dijitCalendarBodyContainer TBODY TR TD {
	CURSOR: hand
}
.dijitCalendarPreviousMonthDisabled {
	CURSOR: default !important
}
.dijitCalendarCurrentMonthDisabled {
	CURSOR: default !important
}
.dijitCalendarNextMonthDisabled {
	CURSOR: default !important
}
.dijitCalendarDateTemplate {
	CURSOR: pointer
}
.dijitCalendarSelectedYear {
	CURSOR: pointer
}
.dijitCalendarNextYear {
	CURSOR: pointer
}
.dijitCalendarPreviousYear {
	CURSOR: pointer
}
.dijitCalendarMonthLabelSpacer {
	POSITION: relative; HEIGHT: 1px; VISIBILITY: hidden; OVERFLOW: hidden
}
.dijitMenu {
	BORDER-BOTTOM: black 1px solid; BORDER-LEFT: black 1px solid; =
BACKGROUND-COLOR: white; BORDER-TOP: black 1px solid; BORDER-RIGHT: =
black 1px solid
}
.dijitMenuTable {
	BORDER-RIGHT-WIDTH: 0px; BACKGROUND-COLOR: white; MARGIN: 1px 0px; =
BORDER-COLLAPSE: collapse; BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: =
0px; BORDER-LEFT-WIDTH: 0px
}
.dijitMenuItem {
	PADDING-BOTTOM: 0.1em; PADDING-LEFT: 0.2em; PADDING-RIGHT: 0.2em; =
WHITE-SPACE: nowrap; PADDING-TOP: 0.1em
}
.dijitMenuItemHover {
	BACKGROUND-COLOR: black; COLOR: white; CURSOR: hand
}
.dijitMenuItemIcon {
	POSITION: relative; BACKGROUND-REPEAT: no-repeat
}
.dijitMenuItemDisabled * {
	CURSOR: default; opacity: 0.3
}
.dj_ie .dijit_a11y .dijitMenuItemDisabled TD {
	FILTER: alpha(opacity=3D35); COLOR: gray !important
}
.dj_ie .dijitMenuItemDisabled * {
	FILTER: alpha(opacity=3D35); COLOR: gray !important
}
.dj_ie .dijitMenuItemDisabled TD {
	FILTER: alpha(opacity=3D35); COLOR: gray !important
}
.dijitMenuItemLabel {
	POSITION: relative; VERTICAL-ALIGN: middle
}
.dijit_a11y .dijitMenuItemHover .dijitMenuItemLabel {
	BORDER-BOTTOM: 1px solid; BORDER-LEFT: 1px solid; BORDER-TOP: 1px =
solid; BORDER-RIGHT: 1px solid
}
.dijit_a11y .dijitMenuItemHover {
	BORDER-BOTTOM: #fff 1px dotted; BORDER-LEFT: #fff 1px dotted; =
BORDER-TOP: #fff 1px dotted; BORDER-RIGHT: #fff 1px dotted
}
.dijit_a11y .dijitMenuExpandInner {
	DISPLAY: block !important
}
.dijitMenuSeparatorTop {
	MARGIN: 3px 0px 0px; HEIGHT: 50%; FONT-SIZE: 1px
}
.dijitMenuSeparatorBottom {
	MARGIN: 0px 0px 3px; HEIGHT: 50%; FONT-SIZE: 1px
}
.dijitTabLabels-top {
	Z-INDEX: 10; TOP: 1px !important
}
.dijitTabLabels-bottom {
	Z-INDEX: 10; MARGIN-TOP: -1px !important
}
.dijitTabLabels-left-h {
	Z-INDEX: 10; MARGIN-RIGHT: -1px !important
}
.dijitTabLabels-right-h {
	Z-INDEX: 10; MARGIN-LEFT: -1px !important
}
.dijitTabPaneWrapper {
	Z-INDEX: 0; CLEAR: both; OVERFLOW: hidden
}
.dijitTab {
	Z-INDEX: 3; POSITION: relative; WHITE-SPACE: nowrap; FLOAT: left; =
CURSOR: pointer
}
.dijitTabContainer .dijitAlignLeft .dijitTab {
	FLOAT: none
}
.dijitTabContainer .dijitAlignRight .dijitTab {
	FLOAT: none
}
.dijitTabInnerDiv {
	POSITION: relative
}
.dijitTab .close {
	DISPLAY: inline-block; FONT-SIZE: small; CURSOR: default
}
.dijitTab .closeText {
	PADDING-BOTTOM: 0px; MARGIN: 0px 2px; PADDING-LEFT: 2px; PADDING-RIGHT: =
2px; DISPLAY: none; PADDING-TOP: 0px
}
.dijit_a11y .dijitTab .closeImage {
	PADDING-BOTTOM: 0px !important; MARGIN: 0px; PADDING-LEFT: 0px =
!important; BOTTOM: 0px !important; PADDING-RIGHT: 0px !important; TOP: =
0px !important; PADDING-TOP: 0px !important
}
.dijit_a11y .closeText {
	DISPLAY: inline; MARGIN-LEFT: 6px
}
.dijit_a11y .closeText:hover {
	BORDER-BOTTOM: thin solid; BORDER-LEFT: thin solid; BORDER-TOP: thin =
solid; BORDER-RIGHT: thin solid
}
.dijit_a11y .dijitTabChecked {
	BORDER-BOTTOM-STYLE: dashed !important; BORDER-RIGHT-STYLE: dashed =
!important; BORDER-TOP-STYLE: dashed !important; BORDER-LEFT-STYLE: =
dashed !important
}
.dijit_a11y .dijitTabInnerDiv {
	BORDER-LEFT-STYLE: none !important
}
.dijitInlineEditor {
	POSITION: relative; VERTICAL-ALIGN: bottom
}
.dj_ie .dijitInlineEditor {
	VERTICAL-ALIGN: middle
}
.dijitInlineValue {
=09
}
.dijitInlineEditor .dijitButtonContainer {
	POSITION: absolute; OVERFLOW: visible; RIGHT: 0px
}
.dijitInlineEditor .saveButton {
=09
}
.dijitInlineEditor .cancelButton {
=09
}
.dijitTreeExpando {
	DISPLAY: inline; FLOAT: left
}
.dijitTreeExpand {
	DISPLAY: inline; FLOAT: left
}
.dijitTreeExpando {
	CLEAR: both
}
.dijitTreeContent {
	CURSOR: default
}
.dijitExpandoText {
	DISPLAY: none
}
.dijit_a11y .dijitExpandoText {
	BORDER-BOTTOM: thin solid; BORDER-LEFT: thin solid; PADDING-LEFT: 10px; =
PADDING-RIGHT: 10px; DISPLAY: inline; FONT-FAMILY: monospace; FLOAT: =
left; BORDER-TOP: thin solid; CURSOR: pointer; BORDER-RIGHT: thin solid
}
.dijitDialog {
	Z-INDEX: 999; POSITION: absolute; PADDING-BOTTOM: 1px; PADDING-LEFT: =
1px; PADDING-RIGHT: 1px; PADDING-TOP: 1px
}
.dijitDialogUnderlayWrapper {
	Z-INDEX: 998; POSITION: absolute; DISPLAY: none; BACKGROUND: none =
transparent scroll repeat 0% 0%; TOP: 0px; LEFT: 0px
}
.dijitDialogUnderlay {
	BACKGROUND: #333333; opacity: 0.6
}
.dj_ie .dijitDialogUnderlay {
	FILTER: alpha(opacity=3D60)
}
#CommentDialog_underlay {
	FILTER: alpha(opacity=3D10); opacity: .1
}
#CommentDialogMultiple_underlay {
	FILTER: alpha(opacity=3D10); opacity: .1
}
.dijit_a11y .dijitInputLayoutContainer {
	BACKGROUND-COLOR: white !important; opacity: 1
}
.dijit_a11y .dijitDialog {
	BACKGROUND-COLOR: white !important; opacity: 1
}
.dijitDialog .closeText {
	POSITION: absolute; DISPLAY: none
}
.dijit_a11y .dijitDialog .closeText {
	DISPLAY: inline
}
.dijitSliderMoveable {
	Z-INDEX: 99; POSITION: absolute !important; DISPLAY: block; =
VERTICAL-ALIGN: middle
}
.dijitSliderMoveableH {
	RIGHT: 0px
}
.dijit_a11y DIV.dijitSliderImageHandle {
	BORDER-BOTTOM: gray 8px solid; POSITION: absolute !important; =
BORDER-LEFT: gray 8px solid; PADDING-BOTTOM: 0px; MARGIN: 0px; =
PADDING-LEFT: 0px; WIDTH: 0px; PADDING-RIGHT: 0px; HEIGHT: 0px; =
BORDER-TOP: gray 8px solid; BORDER-RIGHT: gray 8px solid; PADDING-TOP: =
0px
}
.dijitSliderImageHandle {
	BORDER-BOTTOM: gray 8px solid; POSITION: absolute !important; =
BORDER-LEFT: gray 8px solid; PADDING-BOTTOM: 0px; MARGIN: 0px; =
PADDING-LEFT: 0px; WIDTH: 0px; PADDING-RIGHT: 0px; HEIGHT: 0px; =
BORDER-TOP: gray 8px solid; BORDER-RIGHT: gray 8px solid; PADDING-TOP: =
0px
}
.dijit_a11y .dijitSliderFocused .dijitSliderImageHandle {
	BORDER-BOTTOM: #000 4px solid; BORDER-LEFT: #000 4px solid; WIDTH: 8px; =
HEIGHT: 8px; BORDER-TOP: #000 4px solid; BORDER-RIGHT: #000 4px solid
}
.dijitSliderImageHandleV {
	TOP: -8px; LEFT: -6px
}
.dijitSliderImageHandleH {
	VERTICAL-ALIGN: top; TOP: -5px; LEFT: -8px
}
.dijitSliderBar {
	BORDER-BOTTOM-STYLE: solid; BORDER-BOTTOM-COLOR: black; =
BORDER-RIGHT-STYLE: solid; BORDER-TOP-COLOR: black; BORDER-TOP-STYLE: =
solid; BORDER-RIGHT-COLOR: black; BORDER-LEFT-STYLE: solid; =
BORDER-LEFT-COLOR: black
}
.dijitSliderBarH {
	BORDER-RIGHT-WIDTH: 0px; BORDER-TOP-WIDTH: 1px; BORDER-BOTTOM-WIDTH: =
1px; HEIGHT: 4px; BORDER-LEFT-WIDTH: 0px
}
.dijitSliderBarV {
	BORDER-RIGHT-WIDTH: 1px; WIDTH: 4px; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; BORDER-LEFT-WIDTH: 1px
}
.dijitSliderProgressBar {
	Z-INDEX: 0; BACKGROUND-COLOR: red
}
.dijitSliderProgressBarV {
	POSITION: static !important; TEXT-ALIGN: left; HEIGHT: 0%; =
VERTICAL-ALIGN: top
}
.dijitSliderProgressBarH {
	POSITION: absolute !important; WIDTH: 0%; VERTICAL-ALIGN: middle; =
OVERFLOW: visible
}
.dijitSliderRemainingBar {
	Z-INDEX: -1; BACKGROUND-COLOR: transparent; OVERFLOW: hidden
}
.dijitSliderRemainingBarV {
	TEXT-ALIGN: left; HEIGHT: 100%
}
.dijitSliderRemainingBarH {
	WIDTH: 100% !important
}
.dijitSliderBumper {
	Z-INDEX: -1; OVERFLOW: hidden
}
.dijitSliderBumperV {
	BORDER-RIGHT-WIDTH: 1px; WIDTH: 4px; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; HEIGHT: 8px; BORDER-LEFT-WIDTH: 1px
}
.dijitSliderBumperH {
	BORDER-RIGHT-WIDTH: 0px; WIDTH: 8px; BORDER-TOP-WIDTH: 1px; =
BORDER-BOTTOM-WIDTH: 1px; HEIGHT: 4px; BORDER-LEFT-WIDTH: 0px
}
.dijitSliderBottomBumper {
	BACKGROUND-COLOR: red
}
.dijitSliderLeftBumper {
	BACKGROUND-COLOR: red
}
.dijitSliderTopBumper {
	BACKGROUND-COLOR: transparent
}
.dijitSliderRightBumper {
	BACKGROUND-COLOR: transparent
}
.dijitSliderDecorationH {
	TEXT-ALIGN: center
}
.dijitSlider .dijitSliderButton {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; DISPLAY: block; FONT-FAMILY: monospace; PADDING-TOP: 0px
}
.dijit_a11y .dijitSliderButtonInner {
	VISIBILITY: visible !important
}
.dijitSliderButtonContainer {
	TEXT-ALIGN: center; HEIGHT: 0px
}
.dijitSlider .dijitButtonNode {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; DISPLAY: =
block; PADDING-TOP: 0px
}
.dj_ie .dijitRuleContainer {
	Z-INDEX: -1
}
.dijitRuleContainer {
	POSITION: relative; OVERFLOW: visible
}
.dijitRuleContainerV {
	TEXT-ALIGN: left; LINE-HEIGHT: 0; FLOAT: left; HEIGHT: 100%
}
.dj_opera .dijitRuleContainerV {
	LINE-HEIGHT: 2%
}
.dj_ie .dijitRuleContainerV {
	LINE-HEIGHT: normal
}
.dj_gecko .dijitRuleContainerV {
	MARGIN: 0px 0px 1px
}
.dijitRuleMark {
	BORDER-BOTTOM: black 1px solid; POSITION: absolute; BORDER-LEFT: black =
1px solid; LINE-HEIGHT: 0; HEIGHT: 100%; BORDER-TOP: black 1px solid; =
BORDER-RIGHT: black 1px solid
}
.dijitRuleMarkH {
	WIDTH: 0px; BORDER-TOP-WIDTH: 0px !important; BORDER-BOTTOM-WIDTH: 0px =
!important; BORDER-LEFT-WIDTH: 0px !important
}
.dijitRuleLabelContainer {
	POSITION: absolute
}
.dijitRuleLabelContainerH {
	TEXT-ALIGN: center; DISPLAY: inline-block
}
.dijitRuleLabelH {
	POSITION: relative; LEFT: -50%
}
.dijitRuleMarkV {
	BORDER-RIGHT-WIDTH: 0px !important; WIDTH: 100%; BORDER-BOTTOM-WIDTH: =
0px !important; HEIGHT: 0px; BORDER-LEFT-WIDTH: 0px !important; LEFT: =
0px
}
.dj_ie .dijitRuleLabelContainerV {
	MARGIN-TOP: -0.55em
}
.dijit_a11y .dijitButtonContents .dijitButtonText {
	DISPLAY: inline !important
}
.dijitTextArea {
	WIDTH: 100%; OVERFLOW: auto
}
.dj_ie .dijitTextArea P {
	LINE-HEIGHT: normal !important; MARGIN-TOP: 0px; MARGIN-BOTTOM: 0px
}
.dijitToolbarSeparator {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 1px; WIDTH: 5px; =
PADDING-RIGHT: 1px; HEIGHT: 18px; PADDING-TOP: 0px
}
.dijitIEFixedToolbar {
	POSITION: absolute; ; TOP: =
expression(eval((document.documentElement||document.body).scrollTop))
}
.dijitTimePickerItemInner {
	TEXT-ALIGN: center; PADDING-BOTTOM: 2px; BORDER-RIGHT-WIDTH: 0px; =
PADDING-LEFT: 8px; PADDING-RIGHT: 8px; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; PADDING-TOP: 2px
}
.dijitTimePickerTick {
	BORDER-BOTTOM: #dedede 1px solid; POSITION: relative; COLOR: #dedede; =
BORDER-TOP: #dedede 1px solid
}
.dijitTimePickerTick .dijitTimePickerItemInner {
	FONT-SIZE: 0.25em
}
.dijitTimePickerMarker {
	BORDER-BOTTOM: #999 1px solid; BACKGROUND-COLOR: #ededed; BORDER-TOP: =
#999 1px solid
}
.dijitTimePickerItemHover {
	BORDER-BOTTOM: #333 1px solid; BACKGROUND-COLOR: #808080; COLOR: #fff; =
BORDER-TOP: #333 1px solid; CURSOR: pointer; opacity: 1
}
.dijitTimePickerItemHover.dijitTimePickerMarker {
	FONT-SIZE: 1.3em
}
.dijitTimePickerItemHover .dijitTimePickerItemInner {
	BACKGROUND-COLOR: #808080; DISPLAY: block; FONT-SIZE: 1em; OVERFLOW: =
visible
}
.dijitTimePickerItemSelected {
	BACKGROUND-COLOR: #b7cdee !important; COLOR: #333; FONT-WEIGHT: bold
}
.dijit_a11y .dijitTimePickerItem {
	BORDER-BOTTOM: #333 1px solid
}
.dijitToggleButtonIconChar {
	DISPLAY: none !important
}
.dijit_a11y .dijitToggleButtonIconChar {
	DISPLAY: inline !important
}
.dijit_a11y .dijitToggleButtonIconChar {
	VISIBILITY: hidden
}
.dijit_a11y .dijitToggleButtonChecked .dijitToggleButtonIconChar {
	VISIBILITY: visible !important
}
.dijitArrowButtonChar {
	DISPLAY: none !important
}
.dijit_a11y .dijitArrowButtonChar {
	DISPLAY: inline !important
}
.dijitInputLayoutContainer {
	POSITION: relative; OVERFLOW: hidden
}
.dijitSpinnerButtonContainer {
	POSITION: relative; WIDTH: 18px; FLOAT: right; OVERFLOW: hidden
}
.dijitSpinner .dijitInputLayoutContainer .dijitArrowButton {
	WIDTH: 16px; HEIGHT: 50%; OVERFLOW: hidden
}
.dj_ie .dijitSpinner .dijitInputLayoutContainer .dijitArrowButton {
	; BEHAVIOR: expression(=0A=
		function(self){=0A=
			var e; =0A=
			try{=0A=
				var h =3D self.parentNode.offsetHeight/2;=0A=
				if(h > 1){=0A=
					self.style.height =3D h;=0A=
					self.style.behavior =3D null; =0A=
				}=0A=
			}catch(e){} =0A=
		}(this)=0A=
	)
}
.dijitSpinner .dijitInputLayoutContainer .dijitArrowButtonInner {
	LINE-HEIGHT: 50%; OVERFLOW: hidden
}
.dijit_a11y .dijitSpinner .dijitInputLayoutContainer .dijitArrowButton {
	WIDTH: 100%
}
.dijit_a11y .dijitSpinner .dijitArrowButton .dijitArrowButtonChar {
	FONT-SIZE: 0.4em; VERTICAL-ALIGN: top
}
.dijit_a11y .dijitSpinnerButtonContainer {
	WIDTH: 0.5em; MARGIN-LEFT: 2px; OVERFLOW: visible
}
.dijit_a11y .dijitSpinnerButtonContainer .dijitButtonNode {
	BORDER-BOTTOM: 0px solid; BORDER-LEFT: 1px solid; BORDER-TOP: 1px =
solid; BORDER-RIGHT: 0px solid
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Common.css

.tundra .dojoDndItemBefore {
	BORDER-TOP: #369 2px solid
}
.tundra .dojoDndItemAfter {
	BORDER-BOTTOM: #369 2px solid
}
.tundra .dojoDndItemOver {
	CURSOR: pointer
}
.tundra TABLE.dojoDndAvatar {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; =
BACKGROUND-COLOR: #fff; BORDER-COLLAPSE: collapse; COLOR: black; =
FONT-SIZE: 75%; BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #ccc 1px =
solid; -moz-border-radius: 0
}
.tundra .dojoDndAvatar TD {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; BORDER-LEFT-STYLE: none
}
.tundra .dojoDndAvatar TR {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; BORDER-LEFT-STYLE: none
}
.tundra .dojoDndAvatarHeader TD {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 21px; PADDING-RIGHT: 0px; HEIGHT: =
20px; PADDING-TOP: 0px
}
.tundra .dojoDndAvatarItem TD {
	PADDING-BOTTOM: 2px; PADDING-LEFT: 2px; PADDING-RIGHT: 2px; =
PADDING-TOP: 2px
}
.dojoDndMove.tundra .dojoDndAvatarHeader {
	BACKGROUND-IMAGE: url(images/dndNoMove.png); BACKGROUND-COLOR: #f58383; =
BACKGROUND-REPEAT: no-repeat; BACKGROUND-POSITION: 2px center
}
.dojoDndCopy.tundra .dojoDndAvatarHeader {
	BACKGROUND-IMAGE: url(images/dndNoCopy.png); BACKGROUND-COLOR: #f58383; =
BACKGROUND-REPEAT: no-repeat; BACKGROUND-POSITION: 2px center
}
.dojoDndMove.tundra .dojoDndAvatarCanDrop .dojoDndAvatarHeader {
	BACKGROUND-IMAGE: url(images/dndMove.png); BACKGROUND-COLOR: #97e68d; =
BACKGROUND-REPEAT: no-repeat; BACKGROUND-POSITION: 2px center
}
.dojoDndCopy.tundra .dojoDndAvatarCanDrop .dojoDndAvatarHeader {
	BACKGROUND-IMAGE: url(images/dndCopy.png); BACKGROUND-COLOR: #97e68d; =
BACKGROUND-REPEAT: no-repeat; BACKGROUND-POSITION: 2px center
}
.tundra .dijitContentPaneLoading {
	PADDING-LEFT: 25px; BACKGROUND: url(images/loading.gif) no-repeat left =
center
}
.tundra .dijitContentPaneError {
	PADDING-LEFT: 25px; BACKGROUND: url(images/warning.png) no-repeat left =
center
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/layout/ContentPane.css

.tundra .dijitContentPane {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
PADDING-TOP: 0px
}
.tundra .dijitTabContainerTop-dijitContentPane {
	PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #fff; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; PADDING-TOP: 5px
}
.tundra .dijitTabContainerLeft-dijitContentPane {
	PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #fff; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; PADDING-TOP: 5px
}
.tundra .dijitTabContainerBottom-dijitContentPane {
	PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #fff; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; PADDING-TOP: 5px
}
.tundra .dijitTabContainerRight-dijitContentPane {
	PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #fff; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; PADDING-TOP: 5px
}
.tundra .dijitAccordionContainer-dijitContentPane {
	PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #fff; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; PADDING-TOP: 5px
}
.tundra .dijitSplitContainer-dijitContentPane {
	PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #fff; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; PADDING-TOP: 5px
}
.tundra .dijitBorderContainer-dijitContentPane {
	PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #fff; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; PADDING-TOP: 5px
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/layout/TabContainer.css

.tundra .dijitTabPaneWrapper {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; =
PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
BACKGROUND: #fff; BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #ccc 1px =
solid; PADDING-TOP: 0px
}
.tundra .dijitTab {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; =
PADDING-BOTTOM: 0px; LINE-HEIGHT: normal; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BACKGROUND: url(../images/tabEnabled.png) #e2e2e2 =
repeat-x; BORDER-TOP: #ccc 1px solid; MARGIN-RIGHT: 4px; BORDER-RIGHT: =
#ccc 1px solid; PADDING-TOP: 0px
}
.tundra .dijitTabInnerDiv {
	PADDING-BOTTOM: 5px; PADDING-LEFT: 9px; PADDING-RIGHT: 8px; =
PADDING-TOP: 6px
}
.dijitTabSpacer {
	FONT-SIZE: 1px
}
.tundra .dijitTabChecked {
	BACKGROUND-IMAGE: none; BORDER-BOTTOM-COLOR: #ccc; BACKGROUND-COLOR: =
#fff; BORDER-TOP-COLOR: #ccc; BORDER-RIGHT-COLOR: #ccc; =
BORDER-LEFT-COLOR: #ccc
}
.tundra .dijitTabHover {
	BORDER-TOP-COLOR: #92a0b3; BACKGROUND: url(../images/tabHover.gif) =
#e2e2e2 repeat-x; COLOR: #243c5f; BORDER-RIGHT-COLOR: #92a0b3; =
BORDER-LEFT-COLOR: #92a0b3
}
.tundra .dijitTabCheckedHover {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; BACKGROUND: =
#fff; BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #ccc 1px solid
}
.tundra .dijitTab .dijitClosable .closeImage {
	WIDTH: 12px; BACKGROUND: url(../images/tabClose.png) no-repeat right =
top; HEIGHT: 12px
}
.tundra .dijitTab .dijitTabButtonSpacer {
	WIDTH: 1px; HEIGHT: 12px
}
.tundra .dijitTabContainerTabListNested .dijitTab {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; BACKGROUND: none transparent scroll repeat 0% 0%; =
BORDER-LEFT-STYLE: none; TOP: 0px
}
.tundra .dijitTabContainerTabListNested .dijitTab .dijitTabContent {
=09
}
.tundra .dijitTabContainerTabListNested .dijitTabHover .dijitTabContent =
.tabLabel {
	TEXT-DECORATION: underline
}
.tundra .dijitTabContainerTabListNested .dijitTabChecked =
.dijitTabContent .tabLabel {
	FONT-WEIGHT: bold; TEXT-DECORATION: underline
}
.tundra .dijitTabContainer .dijitTabPaneWrapperNested {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; BORDER-LEFT-STYLE: none
}
.tundra .dijitTab .closeImage {
	BACKGROUND-IMAGE: url(../images/tabClose.png)
}
.dj_ie6 .tundra .dijitTab .closeImage {
	BACKGROUND-IMAGE: url(../images/tabClose.gif)
}
.tundra .dijitTab .closeButton-hover .closeImage {
	BACKGROUND-IMAGE: url(../images/tabCloseHover.png)
}
.dj_ie6 .tundra .dijitTab .closeButton-hover .closeImage {
	BACKGROUND-IMAGE: url(../images/tabCloseHover.gif)
}
.tundra .dijitTabContainerTop-tabs {
	BORDER-BOTTOM-COLOR: #ccc; BORDER-TOP-COLOR: #ccc; MARGIN-BOTTOM: -1px; =
BORDER-RIGHT-COLOR: #ccc; BORDER-LEFT-COLOR: #ccc
}
.tundra .dijitTabContainerTop-container {
	BORDER-TOP-STYLE: none
}
.tundra .dijitTabContainerTop-tabs .dijitTabChecked {
	BORDER-BOTTOM-COLOR: white
}
.tundra .dijitTabContainerTopStrip {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; =
PADDING-LEFT: 3px; BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #ccc 1px =
solid; PADDING-TOP: 2px
}
.tundra .dijitTabContainerTopStrip {
	BACKGROUND: #f2f2f2
}
.tundra .dijitTabContainerBottom-tabs {
	BORDER-BOTTOM-COLOR: #ccc; MARGIN-TOP: -1px; BORDER-TOP-COLOR: #ccc; =
BORDER-RIGHT-COLOR: #ccc; BORDER-LEFT-COLOR: #ccc
}
.tundra .dijitTabContainerBottom-container {
	BORDER-BOTTOM-STYLE: none
}
.tundra .dijitTabContainerBottom-tabs .dijitTabChecked {
	BORDER-TOP-COLOR: white
}
.tundra .dijitTabContainerBottomStrip {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; =
PADDING-BOTTOM: 2px; PADDING-LEFT: 3px; BORDER-TOP: #ccc 1px solid; =
BORDER-RIGHT: #ccc 1px solid
}
.tundra .dijitTabContainerBottomStrip {
	BACKGROUND: #f2f2f2
}
.tundra .dijitTabContainerBottom-spacer {
	BACKGROUND: #fff; HEIGHT: 0px; BORDER-TOP: #ccc 1px solid
}
.tundra .dijitTabContainerTop-spacer {
	BACKGROUND: #fff; HEIGHT: 0px; BORDER-TOP: #ccc 1px solid
}
.tundra .dijitTabContainerRight-tabs {
	BORDER-BOTTOM-COLOR: #ccc; BORDER-TOP-COLOR: #ccc; BORDER-RIGHT-COLOR: =
#ccc; MARGIN-LEFT: -1px; BORDER-LEFT-COLOR: #ccc
}
.tundra .dijitTabContainerRight-container {
	BORDER-RIGHT-STYLE: none
}
.tundra .dijitTabContainerRight-tabs .dijitTabChecked {
	BORDER-LEFT-COLOR: white
}
.tundra .dijitTabContainerRightStrip {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; =
PADDING-RIGHT: 2px; BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #ccc 1px =
solid; PADDING-TOP: 3px
}
.tundra .dijitTabContainerRightStrip {
	BACKGROUND: #f2f2f2
}
.tundra .dijitTabContainerLeft-tabs {
	BORDER-BOTTOM-COLOR: #ccc; BORDER-TOP-COLOR: #ccc; BORDER-RIGHT-COLOR: =
#ccc; BORDER-LEFT-COLOR: #ccc; MARGIN-RIGHT: -1px
}
.tundra .dijitTabContainerLeft-container {
	BORDER-LEFT-STYLE: none
}
.tundra .dijitTabContainerLeft-tabs .dijitTabChecked {
	BORDER-RIGHT-COLOR: white
}
.tundra .dijitTabContainerLeftStrip {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; =
PADDING-LEFT: 2px; BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #ccc 1px =
solid; PADDING-TOP: 3px
}
.tundra .dijitTabContainerLeftStrip {
	BACKGROUND: #f2f2f2
}
.tundra .dijitTabContainerLeft-tabs .dijitTab {
	MARGIN-BOTTOM: 4px; MARGIN-RIGHT: 0px
}
.tundra .dijitTabContainerRight-tabs .dijitTab {
	MARGIN-BOTTOM: 4px; MARGIN-RIGHT: 0px
}
.tundra .dijitTabContainerLeft-spacer {
	BORDER-LEFT: #ccc 1px solid; WIDTH: 0px; BACKGROUND: #fff
}
.tundra .dijitTabContainerRight-spacer {
	BORDER-LEFT: #ccc 1px solid; WIDTH: 0px; BACKGROUND: #fff
}
.tundra .dijitTabContainerTop-dijitContentPane =
.dijitTabContainerTop-tabs {
	BORDER-LEFT: #ccc 0px solid; PADDING-LEFT: 0px; BORDER-TOP: #ccc 0px =
solid; BORDER-RIGHT: #ccc 0px solid; PADDING-TOP: 0px
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/layout/AccordionContainer.css

.tundra .dijitAccordionContainer {
	BORDER-BOTTOM-COLOR: #ccc; BACKGROUND-COLOR: #fff; BORDER-TOP-COLOR: =
#ccc; BORDER-RIGHT-COLOR: #ccc; BORDER-LEFT-COLOR: #ccc
}
.tundra .dijitAccordionTitle {
	PADDING-BOTTOM: 4px; PADDING-LEFT: 8px; PADDING-RIGHT: 4px; BACKGROUND: =
url(../images/titleBar.png) #fafafa repeat-x left bottom; BORDER-TOP: =
#bfbfbf 1px solid; CURSOR: pointer; PADDING-TOP: 4px
}
.tundra .dijitAccordionTitle-hover {
	BACKGROUND: url(../images/accordionItemHover.gif) #f8fafd repeat-x 50% =
bottom
}
.tundra .dijitAccordionTitle-selected {
	BORDER-BOTTOM: #bfbfbf 1px solid; PADDING-BOTTOM: 4px; PADDING-LEFT: =
8px; PADDING-RIGHT: 4px; BACKGROUND: =
url(../images/accordionItemActive.gif) #f9f9f9 repeat-x 50% bottom; =
BORDER-TOP: #aaaaaa 1px solid; CURSOR: default; FONT-WEIGHT: bold; =
PADDING-TOP: 4px
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/layout/SplitContainer.css

.tundra .dijitSplitContainerSizerH {
	BORDER-LEFT: #bfbfbf 1px solid; WIDTH: 7px; BACKGROUND: =
url(../images/splitContainerSizerV.png) #fff repeat-y; BORDER-TOP-WIDTH: =
0px; BORDER-BOTTOM-WIDTH: 0px; BORDER-RIGHT: #bfbfbf 1px solid
}
.tundra .dijitSplitContainerSizerH .thumb {
	WIDTH: 3px; BACKGROUND: url(../images/splitContainerSizerV-thumb.png) =
#ccc no-repeat; HEIGHT: 19px; OVERFLOW: hidden; LEFT: 1px
}
.tundra .dijitSplitContainerSizerV {
	BORDER-BOTTOM: #bfbfbf 1px solid; BORDER-RIGHT-WIDTH: 0px; BACKGROUND: =
url(../images/splitContainerSizerH.png) #fff repeat-x; HEIGHT: 7px; =
BORDER-TOP: #bfbfbf 1px solid; BORDER-LEFT-WIDTH: 0px
}
.tundra .dijitSplitContainerSizerV .thumb {
	WIDTH: 19px; BACKGROUND: url(../images/splitContainerSizerH-thumb.png) =
#ccc no-repeat; HEIGHT: 3px; OVERFLOW: hidden; TOP: 1px
}
.tundra .dijitSplitter {
	BORDER-BOTTOM-COLOR: #bfbfbf; BORDER-TOP-COLOR: #bfbfbf; =
BORDER-RIGHT-COLOR: #bfbfbf; BORDER-LEFT-COLOR: #bfbfbf
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/layout/BorderContainer.css

.tundra .dijitBorderContainer {
	PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #fcfcfc; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; PADDING-TOP: 5px
}
.tundra .dijitSplitContainer-child {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; BORDER-TOP: =
#ccc 1px solid; BORDER-RIGHT: #ccc 1px solid
}
.tundra .dijitBorderContainer-child {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; BORDER-TOP: =
#ccc 1px solid; BORDER-RIGHT: #ccc 1px solid
}
.tundra .dijitBorderContainer-dijitTabContainerTop {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; BORDER-LEFT-STYLE: none
}
.tundra .dijitBorderContainer-dijitTabContainerBottom {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; BORDER-LEFT-STYLE: none
}
.tundra .dijitBorderContainer-dijitTabContainerLeft {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; BORDER-LEFT-STYLE: none
}
.tundra .dijitBorderContainer-dijitTabContainerRight {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; BORDER-TOP-STYLE: =
none; BORDER-LEFT-STYLE: none
}
.tundra .dijitBorderContainer-dijitBorderContainer {
	BORDER-BOTTOM-STYLE: none; PADDING-BOTTOM: 0px; BORDER-RIGHT-STYLE: =
none; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BORDER-TOP-STYLE: none; =
BORDER-LEFT-STYLE: none; PADDING-TOP: 0px
}
.tundra .dijitSplitterH {
	BORDER-RIGHT-WIDTH: 0px; BACKGROUND: #fcfcfc; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; HEIGHT: 5px; BORDER-LEFT-WIDTH: 0px
}
.tundra .dijitGutterH {
	BORDER-RIGHT-WIDTH: 0px; BACKGROUND: #fcfcfc; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; HEIGHT: 5px; BORDER-LEFT-WIDTH: 0px
}
.tundra .dijitSplitterH .dijitSplitterThumb {
	WIDTH: 19px; BACKGROUND: url(../images/splitContainerSizerH-thumb.png) =
no-repeat; HEIGHT: 2px; TOP: 1px
}
.tundra .dijitSplitterV {
	BORDER-RIGHT-WIDTH: 0px; WIDTH: 5px; BACKGROUND: #fcfcfc; =
BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px
}
.tundra .dijitGutterV {
	BORDER-RIGHT-WIDTH: 0px; WIDTH: 5px; BACKGROUND: #fcfcfc; =
BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px
}
.tundra .dijitSplitterV .dijitSplitterThumb {
	WIDTH: 5px; BACKGROUND: url(../images/splitContainerSizerV-thumb.png) =
no-repeat; HEIGHT: 19px; LEFT: 1px
}
.tundra .dijitSplitterActive {
	BACKGROUND-IMAGE: none; FILTER: Alpha(Opacity=3D60); BACKGROUND-COLOR: =
#aaa; MARGIN: 0px; FONT-SIZE: 1px; opacity: 0.6; -moz-opacity: 0.6
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/form/Common.css

.tundra .dijitInputField INPUT {
	MARGIN: 0em 0.1em
}
.tundra .dijitTextBox {
	MARGIN: 0em 0.1em
}
.tundra .dijitComboBox {
	MARGIN: 0em 0.1em
}
.tundra .dijitSpinner {
	MARGIN: 0em 0.1em
}
.tundra .dijitTextBox {
	FONT-FAMILY: sans-serif; FONT-SIZE: 100%
}
.tundra .dijitComboBox {
	FONT-FAMILY: sans-serif; FONT-SIZE: 100%
}
.tundra .dijitSpinner {
	FONT-FAMILY: sans-serif; FONT-SIZE: 100%
}
.tundra .dijitTextArea {
	FONT-FAMILY: sans-serif; FONT-SIZE: 100%
}
.tundra .dijitTextArea {
	PADDING-BOTTOM: 3px; PADDING-LEFT: 3px; PADDING-RIGHT: 3px; =
PADDING-TOP: 3px
}
.tundra .dijitTextBox {
	BORDER-BOTTOM: #b3b3b3 1px solid; BORDER-LEFT: #b3b3b3 1px solid; =
LINE-HEIGHT: normal; BACKGROUND: url(../images/validationInputBg.gif) =
#fff repeat-x left top; BORDER-TOP: #b3b3b3 1px solid; BORDER-RIGHT: =
#b3b3b3 1px solid
}
.tundra .dijitComboBox {
	BORDER-BOTTOM: #b3b3b3 1px solid; BORDER-LEFT: #b3b3b3 1px solid; =
LINE-HEIGHT: normal; BACKGROUND: url(../images/validationInputBg.gif) =
#fff repeat-x left top; BORDER-TOP: #b3b3b3 1px solid; BORDER-RIGHT: =
#b3b3b3 1px solid
}
.tundra .dijitSpinner {
	BORDER-BOTTOM: #b3b3b3 1px solid; BORDER-LEFT: #b3b3b3 1px solid; =
LINE-HEIGHT: normal; BACKGROUND: url(../images/validationInputBg.gif) =
#fff repeat-x left top; BORDER-TOP: #b3b3b3 1px solid; BORDER-RIGHT: =
#b3b3b3 1px solid
}
.tundra .dijitTextArea {
	BORDER-BOTTOM: #b3b3b3 1px solid; BORDER-LEFT: #b3b3b3 1px solid; =
LINE-HEIGHT: normal; BACKGROUND: url(../images/validationInputBg.gif) =
#fff repeat-x left top; BORDER-TOP: #b3b3b3 1px solid; BORDER-RIGHT: =
#b3b3b3 1px solid
}
.tundra .dijitInlineEditor INPUT {
	BORDER-BOTTOM: #b3b3b3 1px solid; BORDER-LEFT: #b3b3b3 1px solid; =
LINE-HEIGHT: normal; BACKGROUND: url(../images/validationInputBg.gif) =
#fff repeat-x left top; BORDER-TOP: #b3b3b3 1px solid; BORDER-RIGHT: =
#b3b3b3 1px solid
}
.tundra .dijitTextBoxDisabled {
	COLOR: gray
}
.tundra .dijitComboBoxDisabled {
	COLOR: gray
}
.tundra .dijitSpinnerDisabled {
	COLOR: gray
}
.tundra .dijitTextAreaDisabled {
	COLOR: gray
}
.dj_webkit .tundra .dijitTextBoxDisabled {
	COLOR: #eee
}
.dj_webkit .tundra .dijitComboBoxDisabled {
	COLOR: #eee
}
.dj_webkit .tundra .dijitSpinnerDisabled {
	COLOR: #eee
}
.dj_webkit .tundra .dijitTextAreaDisabled {
	COLOR: #eee
}
.tundra .dijitComboBox .dijitInputField {
	PADDING-BOTTOM: 1px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
PADDING-TOP: 1px
}
.tundra .dijitComboBox .dijitButtonNode {
	PADDING-BOTTOM: 1px; PADDING-LEFT: 0.2em; PADDING-RIGHT: 0.2em; =
PADDING-TOP: 1px
}
.tundra .dijitComboBox .dijitButtonNode {
	BORDER-BOTTOM: #9b9b9b 0px solid; BORDER-LEFT: #9b9b9b 1px solid; =
BORDER-TOP: #9b9b9b 0px solid; BORDER-RIGHT: #9b9b9b 0px solid
}
.tundra .dijitSpinner .dijitButtonNode {
	BORDER-BOTTOM: #9b9b9b 0px solid; BORDER-LEFT: #9b9b9b 1px solid; =
BORDER-TOP: #9b9b9b 0px solid; BORDER-RIGHT: #9b9b9b 0px solid
}
.tundra .dijitTextBoxFocused {
	BORDER-BOTTOM-COLOR: #406b9b; BORDER-TOP-COLOR: #406b9b; =
BORDER-RIGHT-COLOR: #406b9b; BORDER-LEFT-COLOR: #406b9b
}
.tundra .dijitComboBoxFocused {
	BORDER-BOTTOM-COLOR: #406b9b; BORDER-TOP-COLOR: #406b9b; =
BORDER-RIGHT-COLOR: #406b9b; BORDER-LEFT-COLOR: #406b9b
}
.tundra .dijitSpinnerFocused {
	BORDER-BOTTOM-COLOR: #406b9b; BORDER-TOP-COLOR: #406b9b; =
BORDER-RIGHT-COLOR: #406b9b; BORDER-LEFT-COLOR: #406b9b
}
.tundra .dijitSpinnerUpArrowActive {
	BORDER-BOTTOM-COLOR: #406b9b; BORDER-TOP-COLOR: #406b9b; =
BORDER-RIGHT-COLOR: #406b9b; BORDER-LEFT-COLOR: #406b9b
}
.tundra .dijitSpinnerDownArrowActive {
	BORDER-BOTTOM-COLOR: #406b9b; BORDER-TOP-COLOR: #406b9b; =
BORDER-RIGHT-COLOR: #406b9b; BORDER-LEFT-COLOR: #406b9b
}
.tundra .dijitTextAreaFocused {
	BORDER-BOTTOM-COLOR: #406b9b; BORDER-TOP-COLOR: #406b9b; =
BORDER-RIGHT-COLOR: #406b9b; BORDER-LEFT-COLOR: #406b9b
}
.tundra .dijitComboBoxFocused .dijitButtonNode {
	BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitSpinnerFocused .dijitButtonNode {
	BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitSpinnerUpArrowActive .dijitButtonNode {
	BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitSpinnerDownArrowActive .dijitButtonNode {
	BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitSpinnerFocused .dijitDownArrowButton {
	BORDER-TOP-COLOR: #366dba
}
.tundra .dijitSpinnerUpArrowActive .dijitDownArrowButton {
	BORDER-TOP-COLOR: #366dba
}
.tundra .dijitSpinnerDownArrowActive .dijitDownArrowButton {
	BORDER-TOP-COLOR: #366dba
}
.tundra .dijitError {
	BACKGROUND-IMAGE: none; BACKGROUND-COLOR: #f9f7ba
}
.dj_ie6 .tundra .dijitError INPUT {
	BACKGROUND-COLOR: #f9f7ba !important
}
.tundra .dijitErrorFocused {
	BACKGROUND-IMAGE: none; BACKGROUND-COLOR: #f9f999
}
.dj_ie6 .tundra .dijitErrorFocused INPUT {
	BACKGROUND-COLOR: #f9f999 !important
}
.tundra .dijitValidationIcon {
	WIDTH: 16px; BACKGROUND: url(../images/warning.png) no-repeat center =
center
}
.tundra .dijitInlineValue {
	PADDING-BOTTOM: 3px; MARGIN: 4px; PADDING-LEFT: 3px; PADDING-RIGHT: =
3px; PADDING-TOP: 3px
}
.tundra .dijitInlineEditor {
	DISPLAY: inline
}
.dijitInlineEditor .saveButton {
	MARGIN: 3px 3px 3px 0px
}
.dijitInlineEditor .cancelButton {
	MARGIN: 3px 3px 3px 0px
}
.RichTextEditable {
	BORDER-BOTTOM: #bfbfbf 1px solid; BORDER-LEFT: #bfbfbf 1px solid; =
BORDER-TOP: #bfbfbf 0px solid; BORDER-RIGHT: #bfbfbf 1px solid
}
.tundra .dijitComboBoxHighlightMatch {
	BACKGROUND-COLOR: #a5beda
}
.tundra .dijitFocusedLabel {
	OUTLINE-STYLE: dotted; OUTLINE-COLOR: #666666; OUTLINE-WIDTH: 1px
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/form/Button.css

.tundra .dijitButtonNode {
	BORDER-BOTTOM: #9b9b9b 1px solid; BORDER-LEFT: #c0c0c0 1px solid; =
PADDING-BOTTOM: 0.2em; PADDING-LEFT: 0.2em; PADDING-RIGHT: 0.2em; =
BACKGROUND: url(../images/buttonEnabled.png) #fff repeat-x left bottom; =
BORDER-TOP: #c0c0c0 1px solid; BORDER-RIGHT: #c0c0c0 1px solid; =
PADDING-TOP: 0.1em
}
.tundra .dijitButtonNode BUTTON {
	BORDER-RIGHT-WIDTH: 0px; BACKGROUND-COLOR: transparent; =
BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px
}
.tundra .dijitButtonText {
	TEXT-ALIGN: center; PADDING-BOTTOM: 0px; PADDING-LEFT: 0.3em; =
PADDING-RIGHT: 0.3em; PADDING-TOP: 0px
}
.tundra .dijitDisabled .dijitButtonText {
	COLOR: #7f7f7f
}
.tundra .dijitArrowButton {
	COLOR: #111
}
.tundra .dijitComboButton .dijitDownArrowButton {
	PADDING-RIGHT: 4px
}
.dj_ff2 .tundra .dijitComboButton .dijitButtonContentsFocused {
	BORDER-BOTTOM: transparent 1px solid; BORDER-LEFT: transparent 1px =
solid; BORDER-TOP: transparent 1px solid; BORDER-RIGHT: transparent 1px =
solid
}
.dj_ff2 .tundra .dijitComboButton .dijitDownArrowButtonFocused {
	BORDER-BOTTOM: transparent 1px solid; BORDER-LEFT: transparent 1px =
solid; BORDER-TOP: transparent 1px solid; BORDER-RIGHT: transparent 1px =
solid
}
.tundra .dijitButtonDisabled .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #bdbdbd; BORDER-TOP-COLOR: #d5d5d5; BACKGROUND: =
url(../images/buttonDisabled.png) #e4e4e4 repeat-x 50% top; =
BORDER-RIGHT-COLOR: #d5d5d5; BORDER-LEFT-COLOR: #d5d5d5
}
.tundra .dijitToggleButtonDisabled .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #bdbdbd; BORDER-TOP-COLOR: #d5d5d5; BACKGROUND: =
url(../images/buttonDisabled.png) #e4e4e4 repeat-x 50% top; =
BORDER-RIGHT-COLOR: #d5d5d5; BORDER-LEFT-COLOR: #d5d5d5
}
.tundra .dijitDropDownButtonDisabled .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #bdbdbd; BORDER-TOP-COLOR: #d5d5d5; BACKGROUND: =
url(../images/buttonDisabled.png) #e4e4e4 repeat-x 50% top; =
BORDER-RIGHT-COLOR: #d5d5d5; BORDER-LEFT-COLOR: #d5d5d5
}
.tundra .dijitComboButtonDisabled .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #bdbdbd; BORDER-TOP-COLOR: #d5d5d5; BACKGROUND: =
url(../images/buttonDisabled.png) #e4e4e4 repeat-x 50% top; =
BORDER-RIGHT-COLOR: #d5d5d5; BORDER-LEFT-COLOR: #d5d5d5
}
.tundra .dijitComboBoxDisabled {
	BORDER-BOTTOM-COLOR: #bdbdbd; BORDER-TOP-COLOR: #d5d5d5; BACKGROUND: =
url(../images/buttonDisabled.png) #e4e4e4 repeat-x 50% top; =
BORDER-RIGHT-COLOR: #d5d5d5; BORDER-LEFT-COLOR: #d5d5d5
}
.tundra .dijitSpinnerDisabled {
	BORDER-BOTTOM-COLOR: #bdbdbd; BORDER-TOP-COLOR: #d5d5d5; BACKGROUND: =
url(../images/buttonDisabled.png) #e4e4e4 repeat-x 50% top; =
BORDER-RIGHT-COLOR: #d5d5d5; BORDER-LEFT-COLOR: #d5d5d5
}
.tundra .dijitSpinnerDisabled .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #bdbdbd; BORDER-TOP-COLOR: #d5d5d5; BACKGROUND: =
url(../images/buttonDisabled.png) #e4e4e4 repeat-x 50% top; =
BORDER-RIGHT-COLOR: #d5d5d5; BORDER-LEFT-COLOR: #d5d5d5
}
.tundra .dijitButtonHover .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #5c7590; BORDER-TOP-COLOR: #a5beda; BACKGROUND: =
url(../images/buttonHover.png) #fcfdff repeat-x 50% bottom; COLOR: =
#243c5f; BORDER-RIGHT-COLOR: #a5beda; BORDER-LEFT-COLOR: #a5beda
}
.tundra .dijitButtonNodeHover {
	BORDER-BOTTOM-COLOR: #5c7590; BORDER-TOP-COLOR: #a5beda; BACKGROUND: =
url(../images/buttonHover.png) #fcfdff repeat-x 50% bottom; COLOR: =
#243c5f; BORDER-RIGHT-COLOR: #a5beda; BORDER-LEFT-COLOR: #a5beda
}
.tundra .dijitToggleButtonHover .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #5c7590; BORDER-TOP-COLOR: #a5beda; BACKGROUND: =
url(../images/buttonHover.png) #fcfdff repeat-x 50% bottom; COLOR: =
#243c5f; BORDER-RIGHT-COLOR: #a5beda; BORDER-LEFT-COLOR: #a5beda
}
.tundra .dijitDropDownButtonHover .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #5c7590; BORDER-TOP-COLOR: #a5beda; BACKGROUND: =
url(../images/buttonHover.png) #fcfdff repeat-x 50% bottom; COLOR: =
#243c5f; BORDER-RIGHT-COLOR: #a5beda; BORDER-LEFT-COLOR: #a5beda
}
.tundra .dijitComboButtonHover .dijitButtonContents {
	BORDER-BOTTOM-COLOR: #5c7590; BORDER-TOP-COLOR: #a5beda; BACKGROUND: =
url(../images/buttonHover.png) #fcfdff repeat-x 50% bottom; COLOR: =
#243c5f; BORDER-RIGHT-COLOR: #a5beda; BORDER-LEFT-COLOR: #a5beda
}
.tundra .dijitComboButtonDownArrowHover .dijitDownArrowButton {
	BORDER-BOTTOM-COLOR: #5c7590; BORDER-TOP-COLOR: #a5beda; BACKGROUND: =
url(../images/buttonHover.png) #fcfdff repeat-x 50% bottom; COLOR: =
#243c5f; BORDER-RIGHT-COLOR: #a5beda; BORDER-LEFT-COLOR: #a5beda
}
.tundra .dijitComboBoxHover .dijitDownArrowButton {
	BACKGROUND: url(../images/buttonHover.png) #fcfdff repeat-x 50% bottom; =
COLOR: #243c5f
}
.tundra .dijitSpinnerUpArrowHover .dijitUpArrowButton {
	BACKGROUND: url(../images/buttonHover.png) #fcfdff repeat-x 50% bottom; =
COLOR: #243c5f
}
.tundra .dijitSpinnerDownArrowHover .dijitDownArrowButton {
	BACKGROUND: url(../images/buttonHover.png) #fcfdff repeat-x 50% bottom; =
COLOR: #243c5f
}
.tundra .dijitSpinnerUpArrowActive .dijitUpArrowButton {
	BORDER-BOTTOM-COLOR: #366dba; BORDER-TOP-COLOR: #366dba; BACKGROUND: =
url(../images/buttonActive.png) #ededed repeat-x 50% bottom; =
BORDER-RIGHT-COLOR: #366dba; BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitSpinnerDownArrowActive .dijitDownArrowButton {
	BORDER-BOTTOM-COLOR: #366dba; BORDER-TOP-COLOR: #366dba; BACKGROUND: =
url(../images/buttonActive.png) #ededed repeat-x 50% bottom; =
BORDER-RIGHT-COLOR: #366dba; BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitButtonActive .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #366dba; BORDER-TOP-COLOR: #366dba; BACKGROUND: =
url(../images/buttonActive.png) #ededed repeat-x 50% bottom; =
BORDER-RIGHT-COLOR: #366dba; BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitToggleButtonActive .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #366dba; BORDER-TOP-COLOR: #366dba; BACKGROUND: =
url(../images/buttonActive.png) #ededed repeat-x 50% bottom; =
BORDER-RIGHT-COLOR: #366dba; BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitDropDownButtonActive .dijitButtonNode {
	BORDER-BOTTOM-COLOR: #366dba; BORDER-TOP-COLOR: #366dba; BACKGROUND: =
url(../images/buttonActive.png) #ededed repeat-x 50% bottom; =
BORDER-RIGHT-COLOR: #366dba; BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitComboButtonActive .dijitButtonContents {
	BORDER-BOTTOM-COLOR: #366dba; BORDER-TOP-COLOR: #366dba; BACKGROUND: =
url(../images/buttonActive.png) #ededed repeat-x 50% bottom; =
BORDER-RIGHT-COLOR: #366dba; BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitDownArrowActive .dijitDownArrowButton {
	BORDER-BOTTOM-COLOR: #366dba; BORDER-TOP-COLOR: #366dba; BACKGROUND: =
url(../images/buttonActive.png) #ededed repeat-x 50% bottom; =
BORDER-RIGHT-COLOR: #366dba; BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitComboBoxActive .dijitDownArrowButton {
	BORDER-BOTTOM-COLOR: #366dba; BORDER-TOP-COLOR: #366dba; BACKGROUND: =
url(../images/buttonActive.png) #ededed repeat-x 50% bottom; =
BORDER-RIGHT-COLOR: #366dba; BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitStackController .dijitToggleButtonChecked .dijitButtonNode =
{
	BORDER-BOTTOM-COLOR: #366dba; BORDER-TOP-COLOR: #366dba; BACKGROUND: =
url(../images/buttonActive.png) #ededed repeat-x 50% bottom; =
BORDER-RIGHT-COLOR: #366dba; BORDER-LEFT-COLOR: #366dba
}
.tundra .dijitArrowButtonInner {
	BACKGROUND-IMAGE: url(../images/spriteArrows.png); MARGIN: 0px 4px 0px =
5px; WIDTH: 7px; BACKGROUND-POSITION: left top; HEIGHT: 7px; FONT-SIZE: =
1px
}
.dj_ie6 .tundra .dijitArrowButtonInner {
	BACKGROUND-IMAGE: url(../images/spriteArrows.gif)
}
.tundra .dijitDisabled .dijitArrowButtonInner {
	BACKGROUND-POSITION: -28px top
}
.tundra .dijitUpArrowButton .dijitArrowButtonInner {
	BACKGROUND-POSITION: -21px top
}
.tundra .dijitDisabled .dijitUpArrowButton .dijitArrowButtonInner {
	BACKGROUND-POSITION: -49px top
}
.tundra .dijitComboBox .dijitArrowButton .dijitArrowButtonInner {
	BACKGROUND-IMAGE: url(../images/comboArrowDown.gif); =
BACKGROUND-ATTACHMENT: scroll; BACKGROUND-REPEAT: no-repeat; =
BACKGROUND-POSITION: center center; HEIGHT: auto; FONT-SIZE: 100%
}
.tundra .dijitComboBoxHover .dijitArrowButtonInner {
=09
}
.tundra .dijitSpinner .dijitArrowButtonInner {
	POSITION: absolute; LINE-HEIGHT: 2px; MARGIN-TOP: -4px; DISPLAY: block; =
MARGIN-LEFT: -4px; TOP: 50%; LEFT: 50%
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/form/Checkbox.css

.tundra .dijitToggleButton .dijitCheckBoxIcon {
	BACKGROUND-IMAGE: url(../images/checkmarkNoBorder.png)
}
.dj_ie6 .tundra .dijitToggleButton .dijitCheckBoxIcon {
	BACKGROUND-IMAGE: url(../images/checkmarkNoBorder.gif)
}
.tundra .dijitCheckBox {
	BACKGROUND-IMAGE: url(../images/checkmark.png); PADDING-BOTTOM: 0px; =
MARGIN: 0px 2px 0px 0px; PADDING-LEFT: 0px; WIDTH: 16px; PADDING-RIGHT: =
0px; BACKGROUND-REPEAT: no-repeat; HEIGHT: 16px; PADDING-TOP: 0px
}
.tundra .dijitCheckBoxIcon {
	BACKGROUND-IMAGE: url(../images/checkmark.png); PADDING-BOTTOM: 0px; =
MARGIN: 0px 2px 0px 0px; PADDING-LEFT: 0px; WIDTH: 16px; PADDING-RIGHT: =
0px; BACKGROUND-REPEAT: no-repeat; HEIGHT: 16px; PADDING-TOP: 0px
}
.dj_ie6 .tundra .dijitCheckBox {
	BACKGROUND-IMAGE: url(../images/checkmark.gif)
}
.dj_ie6 .tundra .dijitCheckBoxIcon {
	BACKGROUND-IMAGE: url(../images/checkmark.gif)
}
.tundra .dijitCheckBox {
	BACKGROUND-POSITION: -16px 50%
}
.tundra .dijitToggleButton .dijitCheckBoxIcon {
	BACKGROUND-POSITION: -16px 50%
}
.tundra .dijitCheckBoxChecked {
	BACKGROUND-POSITION: 0px 50%
}
.tundra .dijitToggleButtonChecked .dijitCheckBoxIcon {
	BACKGROUND-POSITION: 0px 50%
}
.tundra .dijitCheckBoxDisabled {
	BACKGROUND-POSITION: -48px 50%
}
.tundra .dijitCheckBoxCheckedDisabled {
	BACKGROUND-POSITION: -32px 50%
}
.tundra .dijitCheckBoxHover {
	BACKGROUND-POSITION: -80px 50%
}
.tundra .dijitCheckBoxCheckedHover {
	BACKGROUND-POSITION: -64px 50%
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/form/RadioButton.css

.tundra .dijitToggleButton .dijitRadio {
	BACKGROUND-IMAGE: url(../images/checkmarkNoBorder.png)
}
.tundra .dijitToggleButton .dijitRadioIcon {
	BACKGROUND-IMAGE: url(../images/checkmarkNoBorder.png)
}
.dj_ie6 .tundra .dijitToggleButton .dijitRadio {
	BACKGROUND-IMAGE: url(../images/checkmarkNoBorder.gif)
}
.dj_ie6 .tundra .dijitToggleButton .dijitRadioIcon {
	BACKGROUND-IMAGE: url(../images/checkmarkNoBorder.gif)
}
.tundra .dijitRadio {
	BACKGROUND-IMAGE: url(../images/checkmark.png); PADDING-BOTTOM: 0px; =
MARGIN: 0px; PADDING-LEFT: 0px; WIDTH: 16px; PADDING-RIGHT: 0px; =
BACKGROUND-REPEAT: no-repeat; HEIGHT: 16px; PADDING-TOP: 0px
}
.tundra .dijitRadioIcon {
	BACKGROUND-IMAGE: url(../images/checkmark.png); PADDING-BOTTOM: 0px; =
MARGIN: 0px; PADDING-LEFT: 0px; WIDTH: 16px; PADDING-RIGHT: 0px; =
BACKGROUND-REPEAT: no-repeat; HEIGHT: 16px; PADDING-TOP: 0px
}
.dj_ie6 .tundra .dijitRadio {
	BACKGROUND-IMAGE: url(../images/checkmark.gif)
}
.dj_ie6 .tundra .dijitRadioIcon {
	BACKGROUND-IMAGE: url(../images/checkmark.gif)
}
.tundra .dijitRadio {
	BACKGROUND-POSITION: -112px 50%
}
.tundra .dijitToggleButton .dijitRadioIcon {
	BACKGROUND-POSITION: -112px 50%
}
.tundra .dijitRadioDisabled {
	BACKGROUND-POSITION: -144px 50%
}
.tundra .dijitRadioHover {
	BACKGROUND-POSITION: -176px 50%
}
.tundra .dijitRadioChecked {
	BACKGROUND-POSITION: -96px 50%
}
.tundra .dijitRadioCheckedHover {
	BACKGROUND-POSITION: -96px 50%
}
.tundra .dijitToggleButtonChecked .dijitRadioIcon {
	BACKGROUND-POSITION: -96px 50%
}
.tundra .dijitRadioCheckedDisabled {
	BACKGROUND-POSITION: -128px 50%
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/form/Slider.css

.tundra .dijitSliderProgressBarH {
	BORDER-BOTTOM-COLOR: #aab0bb; BORDER-TOP-COLOR: #aab0bb; BACKGROUND: =
url(../images/sliderFull.png) #c0c2c5 repeat-x left top; =
BORDER-RIGHT-COLOR: #aab0bb; BORDER-LEFT-COLOR: #aab0bb
}
.tundra .dijitSliderProgressBarV {
	BORDER-BOTTOM-COLOR: #aab0bb; BORDER-TOP-COLOR: #aab0bb; BACKGROUND: =
url(../images/sliderFullVertical.png) #c0c2c5 repeat-y left bottom; =
BORDER-RIGHT-COLOR: #aab0bb; BORDER-LEFT-COLOR: #aab0bb
}
.tundra .dijitSliderFocused .dijitSliderProgressBarH {
	BACKGROUND-IMAGE: url(../images/sliderFullFocus.png)
}
.tundra .dijitSliderFocused .dijitSliderLeftBumper {
	BACKGROUND-IMAGE: url(../images/sliderFullFocus.png)
}
.tundra .dijitSliderFocused .dijitSliderProgressBarV {
	BACKGROUND-IMAGE: url(../images/sliderFullVerticalFocus.png)
}
.tundra .dijitSliderFocused .dijitSliderBottomBumper {
	BACKGROUND-IMAGE: url(../images/sliderFullVerticalFocus.png)
}
.tundra .dijitSliderRemainingBarV {
	BORDER-BOTTOM-COLOR: #b4b4b4; BORDER-TOP-COLOR: #b4b4b4; BACKGROUND: =
url(../images/sliderEmptyVertical.png) #dcdcdc repeat-y left bottom; =
BORDER-RIGHT-COLOR: #b4b4b4; BORDER-LEFT-COLOR: #b4b4b4
}
.tundra .dijitSliderRemainingBarH {
	BORDER-BOTTOM-COLOR: #b4b4b4; BORDER-TOP-COLOR: #b4b4b4; BACKGROUND: =
url(../images/sliderEmpty.png) #dcdcdc repeat-x left top; =
BORDER-RIGHT-COLOR: #b4b4b4; BORDER-LEFT-COLOR: #b4b4b4
}
.tundra .dijitSliderBar {
	BORDER-BOTTOM-STYLE: solid; BORDER-RIGHT-STYLE: solid; OUTLINE-WIDTH: =
1px; BORDER-TOP-STYLE: solid; BORDER-LEFT-STYLE: solid
}
.tundra .dijitSliderFocused .dijitSliderBar {
	BORDER-BOTTOM-COLOR: #888; BORDER-TOP-COLOR: #888; BORDER-RIGHT-COLOR: =
#888; BORDER-LEFT-COLOR: #888
}
.dijit_a11y .dijitSliderProgressBar {
	BACKGROUND-COLOR: #888 !important
}
.tundra .dijitSliderImageHandleH {
	BORDER-RIGHT-WIDTH: 0px; WIDTH: 16px; BACKGROUND: =
url(../images/preciseSliderThumb.png) no-repeat center top; =
BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; HEIGHT: 16px; =
BORDER-LEFT-WIDTH: 0px; CURSOR: pointer
}
.tundra .dijitSliderFocused .dijitSliderImageHandleH {
	BACKGROUND-IMAGE: url(../images/preciseSliderThumbFocus.gif)
}
.dj_ie6 .tundra .dijitSliderImageHandleH {
	BACKGROUND-IMAGE: url(../images/preciseSliderThumb.gif)
}
.tundra .dijitSliderLeftBumper {
	BORDER-BOTTOM-COLOR: #aab0bb; BORDER-TOP-COLOR: #aab0bb; BACKGROUND: =
url(../images/sliderFull.png) #c0c2c5 repeat-x left top; =
BORDER-RIGHT-COLOR: #aab0bb; BORDER-LEFT-COLOR: #aab0bb; =
BORDER-LEFT-WIDTH: 1px
}
.tundra .dijitSliderRightBumper {
	BORDER-BOTTOM-COLOR: #b4b4b4; BORDER-RIGHT-WIDTH: 1px; =
BORDER-TOP-COLOR: #b4b4b4; BACKGROUND: url(../images/sliderEmpty.png) =
#dcdcdc repeat-x left top; BORDER-RIGHT-COLOR: #b4b4b4; =
BORDER-LEFT-COLOR: #b4b4b4
}
.tundra .dijitSliderImageHandleV {
	BORDER-RIGHT-WIDTH: 0px; WIDTH: 16px; BACKGROUND: =
url(../images/sliderThumb.png) no-repeat center center; =
BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; HEIGHT: 16px; =
BORDER-LEFT-WIDTH: 0px; CURSOR: pointer
}
.tundra .dijitSliderFocused .dijitSliderImageHandleV {
	BACKGROUND-IMAGE: url(../images/sliderThumbFocus.png)
}
.dj_ie6 .tundra .dijitSliderFocused .dijitSliderImageHandleV {
	BACKGROUND-IMAGE: url(../images/sliderThumbFocus.gif)
}
.tundra .dijitSliderBottomBumper {
	BORDER-BOTTOM-COLOR: #aab0bb; BORDER-TOP-COLOR: #aab0bb; BACKGROUND: =
url(../images/sliderFullVertical.png) #c0c2c5 repeat-y left bottom; =
BORDER-BOTTOM-WIDTH: 1px; BORDER-RIGHT-COLOR: #aab0bb; =
BORDER-LEFT-COLOR: #aab0bb
}
.tundra .dijitSliderTopBumper {
	BORDER-BOTTOM-COLOR: #b4b4b4; BORDER-TOP-COLOR: #b4b4b4; BACKGROUND: =
url(../images/sliderEmptyVertical.png) #dcdcdc repeat-y left top; =
BORDER-TOP-WIDTH: 1px; BORDER-RIGHT-COLOR: #b4b4b4; BORDER-LEFT-COLOR: =
#b4b4b4
}
.tundra .dijitSliderDecrementIconH {
	BACKGROUND-IMAGE: url(../images/spriteArrows.png); MARGIN: 5px; WIDTH: =
7px; BACKGROUND-REPEAT: no-repeat; HEIGHT: 7px; FONT-SIZE: 1px; CURSOR: =
pointer
}
.tundra .dijitSliderDecrementIconV {
	BACKGROUND-IMAGE: url(../images/spriteArrows.png); MARGIN: 5px; WIDTH: =
7px; BACKGROUND-REPEAT: no-repeat; HEIGHT: 7px; FONT-SIZE: 1px; CURSOR: =
pointer
}
.tundra .dijitSliderIncrementIconH {
	BACKGROUND-IMAGE: url(../images/spriteArrows.png); MARGIN: 5px; WIDTH: =
7px; BACKGROUND-REPEAT: no-repeat; HEIGHT: 7px; FONT-SIZE: 1px; CURSOR: =
pointer
}
.tundra .dijitSliderIncrementIconV {
	BACKGROUND-IMAGE: url(../images/spriteArrows.png); MARGIN: 5px; WIDTH: =
7px; BACKGROUND-REPEAT: no-repeat; HEIGHT: 7px; FONT-SIZE: 1px; CURSOR: =
pointer
}
.dj_ie6 .tundra .dijitSliderDecrementIconH {
	BACKGROUND-IMAGE: url(../images/spriteArrows.gif)
}
.dj_ie6 .tundra .dijitSliderDecrementIconV {
	BACKGROUND-IMAGE: url(../images/spriteArrows.gif)
}
.dj_ie6 .tundra .dijitSliderIncrementIconH {
	BACKGROUND-IMAGE: url(../images/spriteArrows.gif)
}
.dj_ie6 .tundra .dijitSliderIncrementIconV {
	BACKGROUND-IMAGE: url(../images/spriteArrows.gif)
}
.tundra .dijitSliderDecrementIconH {
	BACKGROUND-POSITION: -7px 0px
}
.tundra .dijitSliderIncrementIconH {
	BACKGROUND-POSITION: -14px 0px
}
.tundra .dijitSliderDecrementIconV {
	BACKGROUND-POSITION: 0px 0px
}
.tundra .dijitSliderIncrementIconV {
	BACKGROUND-POSITION: -21px 0px
}
.tundra .dijitSliderButtonInner {
	VISIBILITY: hidden
}
.tundra .dijitSliderDisabled {
	opacity: 0.6
}
.dj_ie .tundra .dijitSliderDisabled {
	FILTER: gray() alpha(opacity=3D40)
}
.dj_ie .tundra .dijitSliderDisabled .dijitRuleContainer {
	FILTER: gray() alpha(opacity=3D40)
}
.dj_ie .tundra .dijitSliderDisabled .dijitSliderRemainingBar {
	FILTER: gray() alpha(opacity=3D40)
}
.dj_ie .tundra .dijitSliderDisabled .dijitSliderProgressBar {
	FILTER: gray() alpha(opacity=3D40)
}
.tundra .dijitSliderDisabled * {
	CURSOR: not-allowed !important
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Tree.css

.tundra .dijitTreeNode {
	BACKGROUND-IMAGE: url(images/i.gif); ZOOM: 1; BACKGROUND-REPEAT: =
repeat-y
}
.tundra .dijitTreeIsLast {
	BACKGROUND: url(images/i_half.gif) no-repeat
}
.tundra .dijitTreeIsRoot {
	BACKGROUND-IMAGE: none; MARGIN-LEFT: 0px
}
.tundra .dijitTreeExpando {
	WIDTH: 18px; HEIGHT: 18px
}
.tundra .dijitTreeRow {
	PADDING-BOTTOM: 2px
}
.tundra .dijitTreeContent {
	MIN-WIDTH: 18px; MIN-HEIGHT: 18px
}
.tundra .dijitTreeExpand {
	WIDTH: 18px; BACKGROUND-REPEAT: no-repeat; HEIGHT: 18px
}
.tundra .dijitTreeNodeEmphasized {
	BACKGROUND-COLOR: highlight; COLOR: highlighttext
}
.tundra .dijitTreeLabelFocused {
	OUTLINE-STYLE: dotted; OUTLINE-COLOR: invert; OUTLINE-WIDTH: 1px
}
.tundra .dijitTreeNodeSelected .dijitTreeLabel {
	BACKGROUND: #e2ebfe
}
.tundra .dijitTreeNodeHover {
	BACKGROUND-IMAGE: url(images/treeHover.png); BACKGROUND-REPEAT: repeat
}
.tundra .dijitTreeExpandoOpened {
	BACKGROUND-IMAGE: url(images/treeExpand_minus.gif)
}
.tundra .dijitTreeExpandoClosed {
	BACKGROUND-IMAGE: url(images/treeExpand_plus.gif)
}
.tundra .dijitTreeExpandoLeaf {
	BACKGROUND-IMAGE: url(images/treeExpand_leaf.gif)
}
.tundra .dijitTreeExpandoLoading {
	BACKGROUND-IMAGE: url(images/treeExpand_loading.gif)
}
.tundra .dijitTreeIcon {
	WIDTH: 16px; HEIGHT: 16px
}
.tundra .dijitFolderOpened {
	BACKGROUND: url(images/folderOpened.gif) no-repeat
}
.tundra .dijitFolderClosed {
	BACKGROUND: url(images/folderClosed.gif) no-repeat
}
.tundra .dijitLeaf {
	BACKGROUND: url(images/leaf.gif) no-repeat
}
.tundra .dijitTreeNode .dojoDndItemBefore {
	BORDER-BOTTOM-STYLE: none; BORDER-TOP-STYLE: none
}
.tundra .dijitTreeNode .dojoDndItemAfter {
	BORDER-BOTTOM-STYLE: none; BORDER-TOP-STYLE: none
}
.tundra .dijitTreeNode .dojoDndItemBefore .dijitTreeContent {
	BORDER-TOP: #369 2px solid
}
.tundra .dijitTreeNode .dojoDndItemAfter .dijitTreeContent {
	BORDER-BOTTOM: #369 2px solid
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/ProgressBar.css

.tundra .dijitProgressBar {
	MARGIN: 2px 0px
}
.tundra .dijitProgressBarEmpty {
	BORDER-BOTTOM-COLOR: #b8b8b8; BORDER-TOP-COLOR: #a2a2a2; BACKGROUND: =
url(images/progressBarEmpty.png) #fff repeat-x center center; =
BORDER-RIGHT-COLOR: #b8b8b8; BORDER-LEFT-COLOR: #a2a2a2
}
.tundra .dijitProgressBarTile {
	BACKGROUND: url(images/progressBarFull.png) #f0f0f0 repeat-x center =
center
}
.tundra .dijitProgressBarFull {
	BORDER-RIGHT: #b8b8b8 1px solid
}
.tundra .dijitProgressBarLabel {
	COLOR: #293a4b
}
.tundra .dijitProgressBarIndeterminate .dijitProgressBarTile {
	BACKGROUND: url(images/progressBarAnim.gif) #cad2de repeat-x center =
center
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/TitlePane.css

.tundra .dijitTitlePaneTitle {
	BORDER-BOTTOM: #bfbfbf 1px solid; BORDER-LEFT: #bfbfbf 1px solid; =
PADDING-BOTTOM: 3px; PADDING-LEFT: 4px; PADDING-RIGHT: 4px; BACKGROUND: =
url(images/titleBar.png) #fff repeat-x left bottom; BORDER-TOP: #bfbfbf =
1px solid; CURSOR: pointer; BORDER-RIGHT: #bfbfbf 1px solid; =
PADDING-TOP: 3px
}
.tundra .dijitTitlePaneTitle-hover {
	BACKGROUND: url(images/accordionItemHover.gif) #f8fafd repeat-x 50% =
bottom
}
.tundra .dijitTitlePaneTitle * {
	VERTICAL-ALIGN: middle
}
.tundra .dijitTitlePane .dijitArrowNode {
	BACKGROUND-IMAGE: url(images/spriteArrows.png); WIDTH: 7px; =
BACKGROUND-REPEAT: no-repeat; HEIGHT: 7px; CURSOR: pointer
}
.dj_ie6 .tundra .dijitTitlePane .dijitArrowNode {
	BACKGROUND-IMAGE: url(images/spriteArrows.gif)
}
.tundra .dijitTitlePane .dijitOpen .dijitArrowNode {
	BACKGROUND-POSITION: 0px 0px
}
.tundra .dijitTitlePane .dijitClosed .dijitArrowNode {
	BACKGROUND-POSITION: -14px 0px
}
.tundra .dijitTitlePaneFocused .dijitTitlePaneTextNode {
	TEXT-DECORATION: underline
}
.tundra .dijitTitlePane .dijitArrowNodeInner {
	DISPLAY: none
}
.dijit_a11y .dijitTitlePane .dijitArrowNodeInner {
	PADDING-BOTTOM: 0.2em; DISPLAY: inline !important; FONT-FAMILY: =
monospace; CURSOR: pointer
}
.dijit_a11y .dijitTitlePane .dijitArrowNode {
	DISPLAY: none
}
.tundra .dijitTitlePaneContentOuter {
	BORDER-BOTTOM: #bfbfbf 1px solid; BORDER-LEFT: #bfbfbf 1px solid; =
BACKGROUND: #ffffff; BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #bfbfbf =
1px solid
}
.tundra .dijitTitlePaneContentInner {
	PADDING-BOTTOM: 10px; PADDING-LEFT: 10px; PADDING-RIGHT: 10px; =
PADDING-TOP: 10px
}
.dj_ie6 .tundra .dijitTitlePaneContentOuter {
	ZOOM: 1
}
.dj_ie6 .tundra .dijitTitlePane .dijitTitlePaneTitle {
	ZOOM: 1
}
.tundra .dijitTitlePaneTextNode {
	MARGIN-LEFT: 4px; MARGIN-RIGHT: 4px
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Calendar.css

.tundra .dijitCalendarIncrementControl {
	BACKGROUND-IMAGE: url(images/spriteRoundedIconsSmall.png); WIDTH: 15px; =
BACKGROUND-REPEAT: no-repeat; HEIGHT: 15px
}
.dj_ie6 .tundra .dijitCalendarIncrementControl {
	BACKGROUND-IMAGE: url(images/spriteRoundedIconsSmall.gif); FONT-SIZE: =
0.1em
}
.tundra .dijitA11ySideArrow {
	DISPLAY: none
}
.tundra .dijitCalendarDecrease {
	BACKGROUND-POSITION: left top
}
.tundra .dijitCalendarIncrease {
	BACKGROUND-POSITION: -30px top
}
.tundra TABLE.dijitCalendarContainer {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; MARGIN: =
0px; BORDER-SPACING: 0; BORDER-COLLAPSE: separate; FONT-SIZE: 100%; =
BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #ccc 1px solid
}
.tundra .dijitCalendarMonthContainer TH {
	TEXT-ALIGN: center; PADDING-BOTTOM: 0.2em; BACKGROUND: =
url(images/titleBar.png) #d3d3d3 repeat-x 50% top; PADDING-TOP: 0.3em
}
.dj_ie6 .tundra .dijitCalendarMonthContainer TH {
	PADDING-BOTTOM: 0.1em; PADDING-TOP: 0.2em
}
.tundra .dijitCalendarDayLabelTemplate {
	TEXT-ALIGN: center; PADDING-BOTTOM: 0em; BACKGROUND: =
url(images/calendarDayLabel.png) white repeat-x 50% bottom; COLOR: =
#293a4b; BORDER-TOP: #eeeeee 1px solid; FONT-WEIGHT: normal; =
PADDING-TOP: 0.15em
}
.tundra .dijitCalendarBodyContainer {
	BORDER-BOTTOM: #eeeeee 1px solid
}
.tundra .dijitCalendarMonthLabel {
	COLOR: #293a4b; FONT-WEIGHT: bold
}
.tundra .dijitCalendarDateTemplate {
	TEXT-ALIGN: center; PADDING-BOTTOM: 0.05em; PADDING-LEFT: 0.3em; =
PADDING-RIGHT: 0.3em; LETTER-SPACING: 1px; FONT-SIZE: 0.9em; =
FONT-WEIGHT: bold; PADDING-TOP: 0.3em
}
.dj_ie .tundra .dijitCalendarDateTemplate {
	PADDING-BOTTOM: 0.02em; PADDING-LEFT: 0.33em; PADDING-RIGHT: 0.33em; =
PADDING-TOP: 0.1em
}
.tundra .dijitCalendarPreviousMonth {
	BACKGROUND-COLOR: #f8f8f8 !important; COLOR: #999999
}
.tundra .dijitCalendarNextMonth {
	BACKGROUND-COLOR: #f8f8f8 !important; COLOR: #999999
}
.tundra .dijitCalendarPreviousMonthDisabled {
	BACKGROUND-COLOR: #a4a5a6 !important
}
.tundra .dijitCalendarNextMonthDisabled {
	BACKGROUND-COLOR: #a4a5a6 !important
}
.tundra .dijitCalendarCurrentMonth {
	BACKGROUND-COLOR: white !important
}
.tundra .dijitCalendarCurrentMonthDisabled {
	BACKGROUND-COLOR: #bbbbbc !important
}
.tundra .dijitCalendarDisabledDate {
	CURSOR: default !important; TEXT-DECORATION: line-through !important
}
.tundra .dijitCalendarCurrentDate {
	FONT-WEIGHT: bold; TEXT-DECORATION: underline
}
.tundra .dijitCalendarHoveredDate {
	BACKGROUND-COLOR: #e2ebf2 !important
}
.tundra .dijitCalendarSelectedDate {
	BACKGROUND-COLOR: #bbc4d0 !important; COLOR: black !important
}
.tundra .dijitCalendarYearContainer {
	BACKGROUND: url(images/calendarYearLabel.png) white repeat-x 50% =
bottom; BORDER-TOP: #ccc 1px solid
}
.tundra .dijitCalendarYearLabel {
	TEXT-ALIGN: center; PADDING-BOTTOM: 0.25em; MARGIN: 0px; PADDING-LEFT: =
0px; PADDING-RIGHT: 0px; PADDING-TOP: 0.4em
}
.tundra .dijitCalendarSelectedYear {
	PADDING-BOTTOM: 0.1em; BACKGROUND-COLOR: #bbc4d0 !important; =
PADDING-LEFT: 0.2em; PADDING-RIGHT: 0.2em; COLOR: black; PADDING-TOP: =
0.2em
}
.tundra .dijitCalendarNextYear {
	COLOR: black !important; FONT-WEIGHT: normal
}
.tundra .dijitCalendarPreviousYear {
	COLOR: black !important; FONT-WEIGHT: normal
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/TimePicker.css

.tundra .dijitTimePickerTick {
	BORDER-BOTTOM-COLOR: #ccc; BORDER-TOP-COLOR: #ccc; BORDER-RIGHT-COLOR: =
#ccc; BORDER-LEFT-COLOR: #ccc
}
.tundra .dijitTimePickerMarker {
	BORDER-BOTTOM-COLOR: #ccc; BORDER-TOP-COLOR: #ccc; BORDER-RIGHT-COLOR: =
#ccc; BORDER-LEFT-COLOR: #ccc
}
.tundra .dijitTimePickerTick {
	COLOR: white
}
.tundra .dijitTimePickerMarker {
	BACKGROUND: url(images/titleBar.png) #d3d3d3 repeat-x 50% top; COLOR: =
#293a4b; FONT-WEIGHT: bold
}
.tundra .dijitTimePickerItemSelected {
	BACKGROUND: #bbc4d0; COLOR: black
}
.tundra .dijitTimePickerItemHover {
	BACKGROUND: #60a1ea; COLOR: white
}
.tundra .dijitTimePickerItemHover {
	Z-INDEX: 10; POSITION: relative
}
.tundra .dijitTimePickerItemSelected {
	Z-INDEX: 10; POSITION: relative
}
.tundra .dijitTimePickerTick .dijitTimePickerItemInner {
	FONT-SIZE: 0.4em
}
.tundra .dijitTimePickerItemHover .dijitTimePickerItemInner {
	FONT-SIZE: 1em
}
.tundra .dijitTimePickerItemSelected .dijitTimePickerItemInner {
	FONT-SIZE: 1em
}
.tundra .dijitTimePickerMarkerHover {
	BORDER-TOP: #ccc 1px solid
}
.tundra .dijitTimePickerTickHover {
	BORDER-BOTTOM-STYLE: none; MARGIN-TOP: -0.3em; MARGIN-BOTTOM: -0.3em
}
.tundra .dijitTimePickerTickSelected {
	BORDER-BOTTOM-STYLE: none; MARGIN-TOP: -0.3em; MARGIN-BOTTOM: -0.3em
}
.dijit_a11y .dijitTimePickerTickHover {
	BORDER-BOTTOM: #333 1px solid; BACKGROUND-COLOR: #ccc !important; =
BORDER-TOP: #333 1px solid
}
.dijit_a11y .dijitTimePickerMarkerHover {
	BORDER-BOTTOM-WIDTH: 2px
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Toolbar.css

.tundra .dijitToolbar {
	BORDER-BOTTOM: #ccc 1px solid; BACKGROUND: url(images/titleBar.png) =
#eaeaea repeat-x left top
}
.dj_ie6 .tundra .dijitToolbar {
	HEIGHT: 10px
}
.tundra .dijitToolbar .dijitButtonNode {
	BORDER-BOTTOM-STYLE: none; PADDING-BOTTOM: 0px; BORDER-RIGHT-STYLE: =
none; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
BORDER-TOP-STYLE: none; BACKGROUND: none transparent scroll repeat 0% =
0%; FONT-SIZE: 12px; BORDER-LEFT-STYLE: none; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitComboButton .dijitButtonContents {
	BORDER-BOTTOM-STYLE: none; PADDING-BOTTOM: 0px; BORDER-RIGHT-STYLE: =
none; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
BORDER-TOP-STYLE: none; BACKGROUND: none transparent scroll repeat 0% =
0%; FONT-SIZE: 12px; BORDER-LEFT-STYLE: none; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitComboButton .dijitDownArrowButton {
	BORDER-BOTTOM-STYLE: none; PADDING-BOTTOM: 0px; BORDER-RIGHT-STYLE: =
none; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
BORDER-TOP-STYLE: none; BACKGROUND: none transparent scroll repeat 0% =
0%; FONT-SIZE: 12px; BORDER-LEFT-STYLE: none; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitButton {
	PADDING-BOTTOM: 1px; PADDING-LEFT: 1px; PADDING-RIGHT: 1px; BACKGROUND: =
none transparent scroll repeat 0% 0%; PADDING-TOP: 1px
}
.tundra .dijitToolbar .dijitToggleButton {
	PADDING-BOTTOM: 1px; PADDING-LEFT: 1px; PADDING-RIGHT: 1px; BACKGROUND: =
none transparent scroll repeat 0% 0%; PADDING-TOP: 1px
}
.tundra .dijitToolbar .dijitDropDownButton {
	PADDING-BOTTOM: 1px; PADDING-LEFT: 1px; PADDING-RIGHT: 1px; BACKGROUND: =
none transparent scroll repeat 0% 0%; PADDING-TOP: 1px
}
.tundra .dijitToolbar .dijitComboButton .dijitButtonContents {
	PADDING-BOTTOM: 1px; PADDING-LEFT: 1px; PADDING-RIGHT: 1px; BACKGROUND: =
none transparent scroll repeat 0% 0%; PADDING-TOP: 1px
}
.tundra .dijitToolbar .dijitComboButton .dijitDownArrowButton {
	PADDING-BOTTOM: 1px; PADDING-LEFT: 1px; PADDING-RIGHT: 1px; BACKGROUND: =
none transparent scroll repeat 0% 0%; PADDING-TOP: 1px
}
.tundra .dijitToolbar .dijitButtonChecked {
	BORDER-BOTTOM: #316ac5 1px solid; BORDER-LEFT: #316ac5 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #d4dff2; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BORDER-TOP: #316ac5 1px solid; BORDER-RIGHT: #316ac5 =
1px solid; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitToggleButtonChecked {
	BORDER-BOTTOM: #316ac5 1px solid; BORDER-LEFT: #316ac5 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #d4dff2; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BORDER-TOP: #316ac5 1px solid; BORDER-RIGHT: #316ac5 =
1px solid; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitButtonCheckedHover {
	BORDER-BOTTOM: #316ac5 1px solid; BORDER-LEFT: #316ac5 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #abc1e5; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BORDER-TOP: #316ac5 1px solid; BORDER-RIGHT: #316ac5 =
1px solid; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitToggleButtonCheckedHover {
	BORDER-BOTTOM: #316ac5 1px solid; BORDER-LEFT: #316ac5 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #abc1e5; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BORDER-TOP: #316ac5 1px solid; BORDER-RIGHT: #316ac5 =
1px solid; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitButtonHover {
	BORDER-BOTTOM: #869cbf 1px solid; BORDER-LEFT: #869cbf 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #e1e5f0; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BORDER-TOP: #869cbf 1px solid; BORDER-RIGHT: #869cbf =
1px solid; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitToggleButtonHover {
	BORDER-BOTTOM: #869cbf 1px solid; BORDER-LEFT: #869cbf 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #e1e5f0; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BORDER-TOP: #869cbf 1px solid; BORDER-RIGHT: #869cbf =
1px solid; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitDropDownButtonHover {
	BORDER-BOTTOM: #869cbf 1px solid; BORDER-LEFT: #869cbf 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #e1e5f0; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BORDER-TOP: #869cbf 1px solid; BORDER-RIGHT: #869cbf =
1px solid; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitComboButtonHover .dijitButtonContents {
	BORDER-BOTTOM: #869cbf 1px solid; BORDER-LEFT: #869cbf 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #e1e5f0; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BORDER-TOP: #869cbf 1px solid; BORDER-RIGHT: #869cbf =
1px solid; PADDING-TOP: 0px
}
.tundra .dijitToolbar .dijitComboButtonDownArrowHover =
.dijitDownArrowButton {
	BORDER-BOTTOM: #869cbf 1px solid; BORDER-LEFT: #869cbf 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #e1e5f0; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BORDER-TOP: #869cbf 1px solid; BORDER-RIGHT: #869cbf =
1px solid; PADDING-TOP: 0px
}
.tundra .dijitToolbar LABEL {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 6px; PADDING-RIGHT: 3px; =
PADDING-TOP: 3px
}
.dj_ie .tundra .dijitToolbar .dijitComboButton =
.dijitButtonContentsFocused {
	BORDER-BOTTOM: #555 1px dotted; BORDER-LEFT: #555 1px dotted; =
PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BORDER-TOP: =
#555 1px dotted; BORDER-RIGHT: #555 1px dotted; PADDING-TOP: 0px
}
.dj_ie .tundra .dijitToolbar .dijitComboButton =
.dijitDownArrowButtonFocused {
	BORDER-BOTTOM: #555 1px dotted; BORDER-LEFT: #555 1px dotted; =
PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BORDER-TOP: =
#555 1px dotted; BORDER-RIGHT: #555 1px dotted; PADDING-TOP: 0px
}
.dj_ff2 .tundra .dijitToolbar .dijitComboButton =
.dijitButtonContentsFocused {
	BORDER-BOTTOM: #555 1px dotted; BORDER-LEFT: #555 1px dotted; =
PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BORDER-TOP: =
#555 1px dotted; BORDER-RIGHT: #555 1px dotted; PADDING-TOP: 0px
}
.dj_ff2 .tundra .dijitToolbar .dijitComboButton =
.dijitDownArrowButtonFocused {
	BORDER-BOTTOM: #555 1px dotted; BORDER-LEFT: #555 1px dotted; =
PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BORDER-TOP: =
#555 1px dotted; BORDER-RIGHT: #555 1px dotted; PADDING-TOP: 0px
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Dialog.css

.tundra .dijitDialog {
	BORDER-BOTTOM: #7eabcd 1px solid; BORDER-LEFT: #7eabcd 1px solid; =
PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BACKGROUND: =
#fff; BORDER-TOP: #7eabcd 1px solid; BORDER-RIGHT: #7eabcd 1px solid; =
PADDING-TOP: 0px; -webkit-box-shadow: 0px 5px 10px #adadad
}
.tundra .dijitDialog .dijitDialogPaneContent {
	BORDER-BOTTOM-STYLE: none; PADDING-BOTTOM: 10px; BORDER-RIGHT-STYLE: =
none; PADDING-LEFT: 10px; PADDING-RIGHT: 10px; BACKGROUND: #fff; =
BORDER-LEFT-STYLE: none; BORDER-TOP: #d3d3d3 1px solid; PADDING-TOP: =
10px
}
.tundra .dijitDialogTitleBar {
	PADDING-BOTTOM: 3px; PADDING-LEFT: 6px; OUTLINE-WIDTH: 0px; =
PADDING-RIGHT: 6px; BACKGROUND: url(images/titleBar.png) #fafafa =
repeat-x left top; CURSOR: move; PADDING-TOP: 5px
}
.tundra .dijitDialogTitle {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 4px; PADDING-RIGHT: 4px; =
FONT-WEIGHT: bold; PADDING-TOP: 0px
}
.tundra .dijitDialogCloseIcon {
	POSITION: absolute; WIDTH: 15px; BACKGROUND: url(images/tabClose.png) =
no-repeat right top; HEIGHT: 15px; VERTICAL-ALIGN: middle; TOP: 4px; =
CURSOR: pointer; RIGHT: 6px
}
.dj_ie6 .tundra .dijitDialogCloseIcon {
	BACKGROUND: url(images/tabClose.gif) no-repeat right top
}
.tundra .dijitDialogCloseIcon-hover {
	BACKGROUND: url(images/tabCloseHover.png) no-repeat right top
}
.dj_ie6 .tundra .dijitDialogCloseIcon-hover {
	BACKGROUND: url(images/tabCloseHover.gif) no-repeat right top
}
.tundra .dijitTooltip {
	BACKGROUND: none transparent scroll repeat 0% 0%
}
.tundra .dijitTooltipDialog {
	BACKGROUND: none transparent scroll repeat 0% 0%
}
.dijitTooltipBelow {
	PADDING-TOP: 13px
}
.dijitTooltipAbove {
	PADDING-BOTTOM: 13px
}
.tundra .dijitTooltipContainer {
	BORDER-BOTTOM: #7eabcd 1px solid; BORDER-LEFT: #7eabcd 1px solid; =
PADDING-BOTTOM: 0.45em; PADDING-LEFT: 0.45em; PADDING-RIGHT: 0.45em; =
BACKGROUND: url(images/popupMenuBg.gif) #ffffff repeat-x left bottom; =
BORDER-TOP: #7eabcd 1px solid; BORDER-RIGHT: #7eabcd 1px solid; =
PADDING-TOP: 0.45em; -webkit-border-radius: 3px
}
.dj_ff3 .tundra .dijitTooltipContainer {
	-moz-border-radius: 3px
}
.tundra .dijitTooltipConnector {
	Z-INDEX: 2; BORDER-RIGHT-WIDTH: 0px; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px
}
.tundra .dijitTooltipABRight .dijitTooltipConnector {
	RIGHT: 3px; LEFT: auto !important
}
.tundra .dijitTooltipBelow .dijitTooltipConnector {
	WIDTH: 16px; BACKGROUND: url(images/tooltipConnectorUp.png) no-repeat =
left top; HEIGHT: 14px; TOP: 0px; LEFT: 3px
}
.dj_ie6 .tundra .dijitTooltipBelow .dijitTooltipConnector {
	BACKGROUND-IMAGE: url(images/tooltipConnectorUp.gif)
}
.tundra .dijitTooltipAbove .dijitTooltipConnector {
	WIDTH: 16px; BOTTOM: 0px; BACKGROUND: =
url(images/tooltipConnectorDown.png) no-repeat left top; HEIGHT: 14px; =
LEFT: 3px
}
.dj_ie6 .tundra .dijitTooltipAbove .dijitTooltipConnector {
	BACKGROUND-IMAGE: url(images/tooltipConnectorDown.gif)
}
.dj_ie6 .tundra .dijitTooltipAbove .dijitTooltipConnector {
	BOTTOM: -3px
}
.tundra .dijitTooltipLeft {
	PADDING-RIGHT: 14px
}
.dj_ie6 .tundra .dijitTooltipLeft {
	PADDING-LEFT: 15px
}
.tundra .dijitTooltipLeft .dijitTooltipConnector {
	WIDTH: 16px; BOTTOM: 3px; BACKGROUND: =
url(images/tooltipConnectorRight.png) no-repeat left top; HEIGHT: 14px; =
RIGHT: 0px
}
.dj_ie6 .tundra .dijitTooltipLeft .dijitTooltipConnector {
	BACKGROUND-IMAGE: url(images/tooltipConnectorRight.gif)
}
.tundra .dijitTooltipRight {
	PADDING-LEFT: 14px
}
.tundra .dijitTooltipRight .dijitTooltipConnector {
	WIDTH: 16px; BOTTOM: 3px; BACKGROUND: =
url(images/tooltipConnectorLeft.png) no-repeat left top; HEIGHT: 14px; =
LEFT: 0px
}
.dj_ie6 .tundra .dijitTooltipRight .dijitTooltipConnector {
	BACKGROUND-IMAGE: url(images/tooltipConnectorLeft.gif)
}
.dj_webkit .tundra .dijitPopup .dijitTooltipContainer {
	-webkit-box-shadow: 0px 5px 10px #adadad
}
.dj_webkit .tundra .dijitPopup .dijitMenu {
	-webkit-box-shadow: 0px 5px 10px #adadad
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Menu.css

.tundra .dijitMenu {
	BORDER-BOTTOM: #7eabcd 1px solid; BORDER-LEFT: #7eabcd 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #f7f7f7; MARGIN: 0px; =
PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BORDER-TOP: #7eabcd 1px solid; =
BORDER-RIGHT: #7eabcd 1px solid; PADDING-TOP: 0px
}
.tundra .dijitMenuBar {
	BORDER-BOTTOM: #7eabcd 1px solid; BORDER-LEFT: #7eabcd 1px solid; =
PADDING-BOTTOM: 0px; BACKGROUND-COLOR: #f7f7f7; MARGIN: 0px; =
PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BORDER-TOP: #7eabcd 1px solid; =
BORDER-RIGHT: #7eabcd 1px solid; PADDING-TOP: 0px
}
.tundra .dijitBorderContainer .dijitMenuBar {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; BORDER-TOP: =
#ccc 1px solid; BORDER-RIGHT: #ccc 1px solid
}
.tundra .dijitMenuItem {
	MARGIN: 0px; FONT-FAMILY: sans-serif
}
.tundra .dijitMenuBar .dijitMenuItem {
	PADDING-BOTTOM: 4px; PADDING-LEFT: 5px; PADDING-RIGHT: 5px; =
PADDING-TOP: 4px
}
.tundra .dijitMenuPreviousButton {
	FONT-STYLE: italic
}
.tundra .dijitMenuNextButton {
	FONT-STYLE: italic
}
.tundra .dijitMenuItem TD {
	PADDING-BOTTOM: 2px; PADDING-LEFT: 2px; PADDING-RIGHT: 2px; =
PADDING-TOP: 2px
}
.tundra .dijitMenuPassive .dijitMenuItemHover {
	BACKGROUND-COLOR: #3559ac; COLOR: #fff
}
.tundra .dijitMenuItemSelected {
	BACKGROUND-COLOR: #3559ac; COLOR: #fff
}
.tundra .dijitMenuItemIcon {
	WIDTH: 16px; HEIGHT: 16px
}
.tundra .dijitMenuExpand {
	BACKGROUND-IMAGE: url(images/spriteArrows.png); WIDTH: 7px; =
BACKGROUND-POSITION: -14px 0px; HEIGHT: 7px
}
.dj_ie6 .tundra .dijitMenuExpand {
	BACKGROUND-IMAGE: url(images/spriteArrows.gif)
}
.tundra .dijitMenuSeparatorTop {
	BORDER-BOTTOM: #9b9b9b 1px solid
}
.tundra .dijitMenuSeparatorBottom {
	BORDER-TOP: #e8e8e8 1px solid
}
.tundra .dijitCheckedMenuItemIconChar {
	DISPLAY: none
}
.tundra .dijitCheckedMenuItemIcon {
	BACKGROUND-IMAGE: url(images/checkmark.png); BACKGROUND-POSITION: -80px =
50%
}
.dj_ie6 .tundra .dijitCheckedMenuItemIcon {
	BACKGROUND-IMAGE: url(images/checkmark.gif)
}
.tundra .dijitCheckedMenuItemChecked .dijitCheckedMenuItemIcon {
	BACKGROUND-POSITION: -64px 50%
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Editor.css

.dijitIEFixedToolbar {
	POSITION: absolute; ; TOP: =
expression(eval((document.documentElement||document.body).scrollTop))
}
.tundra .dijitToolbar .dijitToolbarSeparator {
	BACKGROUND: url(images/editor.gif)
}
.tundra .dijitEditorIcon {
	BACKGROUND-IMAGE: url(images/editor.gif); TEXT-ALIGN: center; WIDTH: =
18px; BACKGROUND-REPEAT: no-repeat; HEIGHT: 18px
}
.tundra .dijitDisabled .dijitEditorIcon {
	BACKGROUND-IMAGE: url(images/editorDisabled.gif)
}
.tundra .dijitEditorIconSep {
	BACKGROUND-POSITION: 0px 50%
}
.tundra .dijitEditorIconBackColor {
	BACKGROUND-POSITION: -18px 50%
}
.tundra .dijitEditorIconBold {
	BACKGROUND-POSITION: -36px 50%
}
.tundra .dijitEditorIconCancel {
	BACKGROUND-POSITION: -54px 50%
}
.tundra .dijitEditorIconCopy {
	BACKGROUND-POSITION: -72px 50%
}
.tundra .dijitEditorIconCreateLink {
	BACKGROUND-POSITION: -90px 50%
}
.tundra .dijitEditorIconCut {
	BACKGROUND-POSITION: -108px 50%
}
.tundra .dijitEditorIconDelete {
	BACKGROUND-POSITION: -126px 50%
}
.tundra .dijitEditorIconForeColor {
	BACKGROUND-POSITION: -144px 50%
}
.tundra .dijitEditorIconHiliteColor {
	BACKGROUND-POSITION: -162px 50%
}
.tundra .dijitEditorIconIndent {
	BACKGROUND-POSITION: -180px 50%
}
.tundra .dijitEditorIconInsertHorizontalRule {
	BACKGROUND-POSITION: -198px 50%
}
.tundra .dijitEditorIconInsertImage {
	BACKGROUND-POSITION: -216px 50%
}
.tundra .dijitEditorIconInsertOrderedList {
	BACKGROUND-POSITION: -234px 50%
}
.tundra .dijitEditorIconInsertTable {
	BACKGROUND-POSITION: -252px 50%
}
.tundra .dijitEditorIconInsertUnorderedList {
	BACKGROUND-POSITION: -270px 50%
}
.tundra .dijitEditorIconItalic {
	BACKGROUND-POSITION: -288px 50%
}
.tundra .dijitEditorIconJustifyCenter {
	BACKGROUND-POSITION: -306px 50%
}
.tundra .dijitEditorIconJustifyFull {
	BACKGROUND-POSITION: -324px 50%
}
.tundra .dijitEditorIconJustifyLeft {
	BACKGROUND-POSITION: -342px 50%
}
.tundra .dijitEditorIconJustifyRight {
	BACKGROUND-POSITION: -360px 50%
}
.tundra .dijitEditorIconLeftToRight {
	BACKGROUND-POSITION: -378px 50%
}
.tundra .dijitEditorIconListBulletIndent {
	BACKGROUND-POSITION: -396px 50%
}
.tundra .dijitEditorIconListBulletOutdent {
	BACKGROUND-POSITION: -414px 50%
}
.tundra .dijitEditorIconListNumIndent {
	BACKGROUND-POSITION: -432px 50%
}
.tundra .dijitEditorIconListNumOutdent {
	BACKGROUND-POSITION: -450px 50%
}
.tundra .dijitEditorIconOutdent {
	BACKGROUND-POSITION: -468px 50%
}
.tundra .dijitEditorIconPaste {
	BACKGROUND-POSITION: -486px 50%
}
.tundra .dijitEditorIconRedo {
	BACKGROUND-POSITION: -504px 50%
}
.tundra .dijitEditorIconRemoveFormat {
	BACKGROUND-POSITION: -522px 50%
}
.tundra .dijitEditorIconRightToLeft {
	BACKGROUND-POSITION: -540px 50%
}
.tundra .dijitEditorIconSave {
	BACKGROUND-POSITION: -558px 50%
}
.tundra .dijitEditorIconSpace {
	BACKGROUND-POSITION: -576px 50%
}
.tundra .dijitEditorIconStrikethrough {
	BACKGROUND-POSITION: -594px 50%
}
.tundra .dijitEditorIconSubscript {
	BACKGROUND-POSITION: -612px 50%
}
.tundra .dijitEditorIconSuperscript {
	BACKGROUND-POSITION: -630px 50%
}
.tundra .dijitEditorIconUnderline {
	BACKGROUND-POSITION: -648px 50%
}
.tundra .dijitEditorIconUndo {
	BACKGROUND-POSITION: -666px 50%
}
.tundra .dijitEditorIconWikiword {
	BACKGROUND-POSITION: -684px 50%
}
.tundra .dijitEditorIconToggleDir {
	BACKGROUND-POSITION: -540px 50%
}
.tundra .dijitEditorIconTabIndent {
	BACKGROUND-POSITION: -702px 50%
}
.tundra .dijitEditorIconSelectAll {
	BACKGROUND-POSITION: -720px 50%
}
.tundra .dijitEditorIconUnlink {
	BACKGROUND-POSITION: -738px 50%
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/ColorPalette.css

.dijitColorPalette {
	BORDER-BOTTOM: #7eabcd 1px solid; BORDER-LEFT: #7eabcd 1px solid; =
BACKGROUND: #fff; BORDER-TOP: #7eabcd 1px solid; BORDER-RIGHT: #7eabcd =
1px solid; -moz-border-radius: 0px
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: 7bit
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Calendar_rtl.css

.dijitRtl .tundra .dijitCalendarDecrease {
	BACKGROUND-POSITION: -30px top
}
.dijitRtl .tundra .dijitCalendarIncrease {
	BACKGROUND-POSITION: 0px top
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: 7bit
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Dialog_rtl.css

.dijitRtl .tundra .dijitDialogTitleBar .dijitDialogCloseIcon {
	RIGHT: auto; LEFT: 5px
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: 7bit
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Editor_rtl.css

.dijitRtl .tundra .dijitEditorIcon {
	BACKGROUND-IMAGE: url(images/editor_rtl.gif)
}
.dijitRtl .tundra .dijitDisabled .dijitEditorIcon {
	BACKGROUND-IMAGE: url(images/editorDisabled_rtl.gif)
}
.dijitRtl .tundra .dijitToolbar .dijitToolbarSeparator {
	BACKGROUND-IMAGE: url(images/editor_rtl.gif)
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: 7bit
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Menu_rtl.css

.dijitRtl .tundra .dijitMenuItem .dijitMenuItemIcon {
	PADDING-LEFT: 3px; PADDING-RIGHT: 0px
}
.dijitRtl .tundra .dijitMenuItem .dijitMenuExpand {
	BACKGROUND-POSITION: -7px 0px
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: 7bit
Content-Location: http://www.plosone.org/article/info:doi/10.1371/Tree_rtl.css

.dijitRtl .tundra .dijitTreeContainer .dijitTreeNode {
	BACKGROUND-IMAGE: none
}
.dijitRtl .tundra .dijitTreeContainer .dijitTreeExpandoOpened {
	BACKGROUND-IMAGE: url(images/treeExpand_minus_rtl.gif)
}
.dijitRtl .tundra .dijitTreeContainer .dijitTreeExpandoClosed {
	BACKGROUND-IMAGE: url(images/treeExpand_plus_rtl.gif)
}
.dijitRtl .tundra .dijitTreeContainer .dijitTreeExpandoLeaf {
	BACKGROUND-IMAGE: none
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: 7bit
Content-Location: http://www.plosone.org/article/info:doi/10.1371/TitlePane_rtl.css

.dijitRtl .tundra .dijitTitlePane .dijitClosed .dijitArrowNode {
	BACKGROUND-POSITION: -7px 0px
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/layout/TabContainer_rtl.css

.dijitRtl .tundra .dijitTab {
	TEXT-ALIGN: right; MARGIN-LEFT: 4px; MARGIN-RIGHT: auto; =
-moz-box-orient: horizontal
}
.dijitRtl .tundra .dijitTabContainerTopStrip {
	PADDING-LEFT: 0px; PADDING-RIGHT: 3px
}
.dijitRtl .tundra .dijitTabContainerBottomStrip {
	PADDING-LEFT: 0px; PADDING-RIGHT: 3px
}
.dijitRtl .tundra .dijitTabInnerDiv {
	PADDING-BOTTOM: 5px; PADDING-LEFT: 8px; PADDING-RIGHT: 9px; =
PADDING-TOP: 6px
}
.dijitRtl .tundra .dijitTabPaneWrapper {
	ZOOM: 1
}
.dj_ie-rtl .tundra .dijitTabContainerLeft-tabs {
	MARGIN-LEFT: 1px !important
}
.dj_ie-rtl .tundra .dijitTabContainerRight-tabs {
	MARGIN-RIGHT: 1px !important
}
.dijitRtl .tundra .dijitTabContainerLeft-tabs .dijitTab {
	MARGIN-LEFT: 0px
}
.dijitRtl .tundra .dijitTabContainerRight-tabs .dijitTab {
	MARGIN-LEFT: 0px
}
.dj_ie-rtl .tundra .dijitTab .dijitTabInnerDiv {
	WIDTH: 1% !important
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: 7bit
Content-Location: http://www.plosone.org/article/info:doi/10.1371/form/Slider_rtl.css

.dijitRtl .tundra .dijitSliderProgressBarH {
	BACKGROUND-POSITION: right top
}
.dijitRtl .tundra .dijitSliderRemainingBarH {
	BACKGROUND-POSITION: right top
}
.dijitRtl .tundra .dijitSliderLeftBumper {
	BACKGROUND-POSITION: right top
}
.dijitRtl .tundra .dijitSliderRightBumper {
	BACKGROUND-POSITION: right top
}
.dijitRtl .tundra .dijitSliderTopBumper {
	BACKGROUND-POSITION: right top
}
.dijitRtl .tundra .dijitSliderProgressBarV {
	BACKGROUND-POSITION: right bottom
}
.dijitRtl .tundra .dijitSliderRemainingBarV {
	BACKGROUND-POSITION: right bottom
}
.dijitRtl .tundra .dijitSliderBottomBumper {
	BACKGROUND-POSITION: right bottom
}
.dijitRtl .tundra .dijitSliderLeftBumper {
	BORDER-RIGHT-WIDTH: 1px; BORDER-LEFT-WIDTH: 0px
}
.dijitRtl .tundra .dijitSliderRightBumper {
	BORDER-RIGHT-WIDTH: 0px; BORDER-LEFT-WIDTH: 1px
}
.dijitRtl .tundra .dijitSliderIncrementIconH {
	BACKGROUND-POSITION: -7px 0px
}
.dijitRtl .tundra .dijitSliderDecrementIconH {
	BACKGROUND-POSITION: -14px 0px
}

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: 7bit
Content-Location: http://www.plosone.org/css/dojo/dijit/themes/tundra/tundra.css

@import url( ../dijit.css );
@import url( Common.css );
@import url( layout/ContentPane.css );
@import url( layout/TabContainer.css );
@import url( layout/AccordionContainer.css );
@import url( layout/SplitContainer.css );
@import url( layout/BorderContainer.css );
@import url( form/Common.css );
@import url( form/Button.css );
@import url( form/Checkbox.css );
@import url( form/RadioButton.css );
@import url( form/Slider.css );
@import url( Tree.css );
@import url( ProgressBar.css );
@import url( TitlePane.css );
@import url( Calendar.css );
@import url( TimePicker.css );
@import url( Toolbar.css );
@import url( Dialog.css );
@import url( Menu.css );
@import url( Editor.css );
@import url( ColorPalette.css );
@import url( ../dijit_rtl.css );
@import url( Calendar_rtl.css );
@import url( Dialog_rtl.css );
@import url( Editor_rtl.css );
@import url( Menu_rtl.css );
@import url( Tree_rtl.css );
@import url( TitlePane_rtl.css );
@import url( layout/TabContainer_rtl.css );
@import url( form/Slider_rtl.css );

------=_NextPart_000_0002_01CA6936.21CCA290
Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/css/star_rating.css

.star-rating {
	BACKGROUND: repeat-x left -1000px
}
.star-rating A:hover {
	BACKGROUND: repeat-x left -1000px
}
.star-rating A:active {
	BACKGROUND: repeat-x left -1000px
}
.star-rating A:focus {
	BACKGROUND: repeat-x left -1000px
}
.star-rating .current-rating {
	BACKGROUND: repeat-x left -1000px
}
.star-rating {
	POSITION: relative; PADDING-BOTTOM: 0px; LIST-STYLE-TYPE: none; MARGIN: =
0px; PADDING-LEFT: 0px; WIDTH: 125px; PADDING-RIGHT: 0px; =
BACKGROUND-POSITION: left top; HEIGHT: 25px; OVERFLOW: hidden; =
PADDING-TOP: 0px
}
.star-rating LI {
	DISPLAY: inline
}
.star-rating A {
	POSITION: absolute; BORDER-BOTTOM-STYLE: none; LINE-HEIGHT: 25px; =
BORDER-RIGHT-STYLE: none; TEXT-INDENT: -1000em; OUTLINE-STYLE: none; =
BORDER-TOP-STYLE: none; HEIGHT: 25px; BORDER-LEFT-STYLE: none; OVERFLOW: =
hidden; TOP: 0px; LEFT: 0px
}
.star-rating .current-rating {
	POSITION: absolute; BORDER-BOTTOM-STYLE: none; LINE-HEIGHT: 25px; =
BORDER-RIGHT-STYLE: none; TEXT-INDENT: -1000em; OUTLINE-STYLE: none; =
BORDER-TOP-STYLE: none; HEIGHT: 25px; BORDER-LEFT-STYLE: none; OVERFLOW: =
hidden; TOP: 0px; LEFT: 0px
}
.star-rating A:hover {
	BACKGROUND-POSITION: left bottom
}
.star-rating A:active {
	BACKGROUND-POSITION: left bottom
}
.star-rating A:focus {
	BACKGROUND-POSITION: left bottom
}
.star-rating A.one-star {
	Z-INDEX: 6; WIDTH: 20%
}
.star-rating A.two-stars {
	Z-INDEX: 5; WIDTH: 40%
}
.star-rating A.three-stars {
	Z-INDEX: 4; WIDTH: 60%
}
.star-rating A.four-stars {
	Z-INDEX: 3; WIDTH: 80%
}
.star-rating A.five-stars {
	Z-INDEX: 2; WIDTH: 100%
}
.star-rating .current-rating {
	Z-INDEX: 1; BACKGROUND-POSITION: left center
}
.inline-rating {
	DISPLAY: inline-block; VERTICAL-ALIGN: middle
}
.small-star {
	WIDTH: 50px; HEIGHT: 10px
}
.small-star {
	LINE-HEIGHT: 10px; HEIGHT: 10px
}
.small-star A:hover {
	LINE-HEIGHT: 10px; HEIGHT: 10px
}
.small-star A:active {
	LINE-HEIGHT: 10px; HEIGHT: 10px
}
.small-star A:focus {
	LINE-HEIGHT: 10px; HEIGHT: 10px
}
.small-star .current-rating {
	LINE-HEIGHT: 10px; HEIGHT: 10px
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/css/rating.css

#ratingAverages {
	DISPLAY: none
}
.rating {
	WIDTH: 100px; HEIGHT: 15px; OVERFLOW: hidden; TOP: -1.25em; =
FONT-WEIGHT: normal; LEFT: 8em
}
OL.ratingAvgs UL.single {
	LEFT: 0px !important
}
SPAN.inline-rating UL.rating {
	TOP: 0px; LEFT: 0px
}
*:first-child + HTML SPAN.inline-rating UL.rating {
	TOP: -1.25em; LEFT: 8em
}
.rating {
	BACKGROUND-IMAGE: url(../images/rating.gif); LINE-HEIGHT: 15px; HEIGHT: =
15px
}
.rating A:hover {
	BACKGROUND-IMAGE: url(../images/rating.gif); LINE-HEIGHT: 15px; HEIGHT: =
15px
}
.rating A:active {
	BACKGROUND-IMAGE: url(../images/rating.gif); LINE-HEIGHT: 15px; HEIGHT: =
15px
}
.rating A:focus {
	BACKGROUND-IMAGE: url(../images/rating.gif); LINE-HEIGHT: 15px; HEIGHT: =
15px
}
.rating .current-rating {
	BACKGROUND-IMAGE: url(../images/rating.gif); LINE-HEIGHT: 15px; HEIGHT: =
15px
}
.rating .current-rating {
	DISPLAY: block; BACKGROUND-POSITION: left bottom
}
.rating .average-rating {
	LEFT: -20px
}
.rating .overall-rating {
	PADDING-LEFT: 0px; LEFT: -20px
}
.rating .single-rating {
	PADDING-LEFT: 0px; LEFT: -20px
}
.rating .pct0 {
	WIDTH: 0%
}
.rating .pct10 {
	WIDTH: 10%
}
.rating .pct20 {
	WIDTH: 20%
}
.rating .pct30 {
	WIDTH: 30%
}
.rating .pct40 {
	WIDTH: 40%
}
.rating .pct50 {
	WIDTH: 50%
}
.rating .pct60 {
	WIDTH: 60%
}
.rating .pct70 {
	WIDTH: 70%
}
.rating .pct80 {
	WIDTH: 80%
}
.rating .pct90 {
	WIDTH: 90%
}
.rating .pct100 {
	WIDTH: 100%
}
SPAN.inlineRatingEnd {
	POSITION: relative; TOP: 0px !important; LEFT: 0px !important
}
*:first-child + HTML SPAN.inlineRatingEnd {
	BACKGROUND: none transparent scroll repeat 0% 0%; TOP: -1.25em; LEFT: =
8em
}
DIV.ratingComment {
	MARGIN: 20px 0px 5px; BORDER-TOP-WIDTH: 1px
}
DIV.ratingComment DIV.hd {
	BACKGROUND: url(../images/toolbar_bg.gif) repeat-x 50% bottom
}
DIV.ratingComment DIV.toolbar A {
	CURSOR: pointer
}
DIV.ratingComment DIV.ratingDetail {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 2px solid; =
PADDING-BOTTOM: 0px; FONT-STYLE: normal; MARGIN: 0px; PADDING-LEFT: =
10px; PADDING-RIGHT: 10px; BACKGROUND: #fff; BORDER-TOP: #ccc 1px solid; =
BORDER-RIGHT: #ccc 0px solid; PADDING-TOP: 5px
}
DIV.hd OL.ratingAvgs {
	MARGIN: 0px
}
DIV.hd OL.ratingAvgs LI {
	PADDING-LEFT: 0px
}
DIV.hd OL.ratingAvgs LABEL {
	FONT-SIZE: 90%
}
DIV.hd OL.ratingAvgs LI UL {
	TOP: -17px; LEFT: 20px
}
DIV.hd SPAN.detail {
	PADDING-LEFT: 5px; FONT-SIZE: 95%; FONT-WEIGHT: normal
}
DIV.hd SPAN.detail A {
	FONT-WEIGHT: bold
}
DIV.ratingDetail OL.ratingAvgs LI {
	PADDING-LEFT: 0px
}
DIV.ratingDetail OL.ratingAvgs LABEL {
	FONT-SIZE: 90%
}
DIV.ratingDetail OL.ratingAvgs LI UL {
	TOP: -17px; LEFT: 7em
}
DIV.ratingDetail BLOCKQUOTE {
	PADDING-BOTTOM: 0px !important; BORDER-RIGHT-WIDTH: 0px !important; =
MARGIN: 0px; PADDING-LEFT: 0px !important; PADDING-RIGHT: 0px =
!important; BACKGROUND: none transparent scroll repeat 0% 0%; =
BORDER-TOP-WIDTH: 0px !important; BORDER-BOTTOM-WIDTH: 0px !important; =
BORDER-LEFT-WIDTH: 0px !important; PADDING-TOP: 0px !important
}
DIV.rsep {
	BORDER-BOTTOM-STYLE: none; PADDING-BOTTOM: 0px; BORDER-RIGHT-STYLE: =
none; MARGIN-TOP: 10px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; =
BORDER-TOP-STYLE: none; BACKGROUND: #fff; BORDER-LEFT-STYLE: none; =
MARGIN-RIGHT: -5px; PADDING-TOP: 0px
}
.inline-rating-rhc {
	POSITION: static; FLOAT: left
}
.inline-rating-rhc .current-rating {
	POSITION: static; FLOAT: none
}
.inline-rating-rhc .overall-rating {
	MARGIN-LEFT: 0px
}
.inline-rating-rhc .single-rating {
	MARGIN-LEFT: 0px
}

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Content-Type: text/css;
	charset="iso-8859-1"
Content-Transfer-Encoding: quoted-printable
Content-Location: http://www.plosone.org/article/info:doi/10.1371/annotation.css

DIV.detail {
	POSITION: relative; COLOR: #666; FONT-SIZE: 0.9em
}
DIV.detail STRONG {
	COLOR: #444
}
.dialog .tip {
	DISPLAY: none
}
.dialog .tipu {
	DISPLAY: none
}
#AnnotationDialog .dialog .tip {
	POSITION: absolute; WIDTH: 39px; BOTTOM: -22px; BACKGROUND: =
url(../images/dialog_pb.gif) no-repeat 0px 0px; HEIGHT: 22px; LEFT: 22px
}
#AnnotationDialog .dialog .tipu {
	POSITION: absolute; WIDTH: 39px; BACKGROUND: =
url(../images/dialog_pt.gif) no-repeat 0px 0px; HEIGHT: 22px; TOP: =
-22px; LEFT: 22px
}
#CommentDialog .dialog .tip {
	POSITION: absolute; WIDTH: 39px; BOTTOM: -18px; BACKGROUND: =
url(../images/dialog_preview_pb.gif) no-repeat 0px 0px; HEIGHT: 22px; =
LEFT: 22px
}
#CommentDialogMultiple .dialog .tip {
	POSITION: absolute; WIDTH: 39px; BOTTOM: -18px; BACKGROUND: =
url(../images/dialog_preview_pb.gif) no-repeat 0px 0px; HEIGHT: 22px; =
LEFT: 22px
}
#CommentDialog .dialog .tipu {
	POSITION: absolute; WIDTH: 39px; BACKGROUND: =
url(../images/dialog_preview_pt.gif) no-repeat 0px 0px; HEIGHT: 22px; =
TOP: -18px; LEFT: 22px
}
#CommentDialogMultiple .dialog .tipu {
	POSITION: absolute; WIDTH: 39px; BACKGROUND: =
url(../images/dialog_preview_pt.gif) no-repeat 0px 0px; HEIGHT: 22px; =
TOP: -18px; LEFT: 22px
}
#CommentDialogMultiple .dialog .tip {
	BOTTOM: -21px
}
#CommentDialogMultiple .dialog .tipu {
	TOP: -21px
}
.dialog .on.tip {
	DISPLAY: block
}
.dialog .on.tipu {
	DISPLAY: block
}
.dialog {
	LINE-HEIGHT: 1.4em; FONT-SIZE: 0.9em
}
.dialog H5 {
	MARGIN: 5px 5px 10px
}
.dialog H6 {
	BORDER-BOTTOM: #666 0px solid; BORDER-LEFT: #666 1px solid; =
PADDING-BOTTOM: 5px; MARGIN: 0px; PADDING-LEFT: 5px; PADDING-RIGHT: 5px; =
BACKGROUND: #fff; CLEAR: right; FONT-SIZE: 1em; BORDER-TOP: #666 1px =
solid; BORDER-RIGHT: #666 1px solid; PADDING-TOP: 5px
}
.dialog H6 A.discuss {
	BACKGROUND: url(../images/icon_annotation.gif) no-repeat left 0.2em
}
DIV.dialog DIV.detail {
	BORDER-BOTTOM: #0bf 1px solid; BORDER-LEFT: #666 1px solid; =
PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #fff; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; BORDER-TOP: #0bf 1px solid; BORDER-RIGHT: #666 1px =
solid; PADDING-TOP: 5px
}
DIV.dialog DIV#viewLink {
	BORDER-BOTTOM-COLOR: #666; BORDER-RIGHT-WIDTH: 1px; BORDER-TOP-COLOR: =
#666; BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 1px; =
BORDER-RIGHT-COLOR: #666; BORDER-LEFT-COLOR: #666; BORDER-LEFT-WIDTH: =
1px
}
DIV.dialog DIV.detail A:link {
	MARGIN: 0px
}
DIV.contentwrap {
	BORDER-BOTTOM: #666 0px solid; BORDER-LEFT: #666 1px solid; =
PADDING-BOTTOM: 10px; BACKGROUND-COLOR: #fff; MARGIN: 0px; PADDING-LEFT: =
10px; PADDING-RIGHT: 10px; BORDER-TOP: #666 0px solid; BORDER-RIGHT: =
#666 1px solid; PADDING-TOP: 10px
}
DIV.contentwrap P {
	MARGIN: 0px
}
DIV.retractionCssStyle#cmtContainer H6 A {
	BACKGROUND: url(../images/icon_correction.gif) no-repeat left 0%; =
COLOR: #ff0000 !important
}
DIV.frmlcrctn#cmtContainer H6 A {
	BACKGROUND: url(../images/icon_correction.gif) no-repeat left 0%; =
COLOR: #ff0000 !important
}
DIV.minrcrctn#cmtContainer H6 A {
	BACKGROUND: url(../images/icon_correction.gif) no-repeat left 0%; =
COLOR: #ff0000 !important
}
DIV.retractionCssStyle#cmtContainer DIV.contentwrap {
	BACKGROUND-COLOR: #fff4f0
}
DIV.frmlcrctn#cmtContainer DIV.contentwrap {
	BACKGROUND-COLOR: #fff4f0
}
DIV.retractionCssStyle#cmtContainer H6#viewCmtTitle {
	BACKGROUND-COLOR: #fff4f0
}
DIV.frmlcrctn#cmtContainer H6#viewCmtTitle {
	BACKGROUND-COLOR: #fff4f0
}
DIV.minrcrctn#cmtContainer DIV.contentwrap {
	BACKGROUND-COLOR: #fff4f0
}
DIV.minrcrctn#cmtContainer H6#viewCmtTitle {
	BACKGROUND-COLOR: #fff4f0
}
DIV.retractionCssStyle#cmtContainer DIV#viewCmtDetail {
	BACKGROUND-COLOR: #fff4f0
}
DIV.frmlcrctn#cmtContainer DIV#viewCmtDetail {
	BACKGROUND-COLOR: #fff4f0
}
DIV.retractionCssStyle#cmtContainer DIV#viewComment {
	BACKGROUND-COLOR: #fff4f0
}
DIV.frmlcrctn#cmtContainer DIV#viewComment {
	BACKGROUND-COLOR: #fff4f0
}
DIV.minrcrctn#cmtContainer DIV#viewCmtDetail {
	BACKGROUND-COLOR: #fff4f0
}
DIV.minrcrctn#cmtContainer DIV#viewComment {
	BACKGROUND-COLOR: #fff4f0
}
DIV.retractionCssStyle#cmtContainer DIV#viewLink {
	BACKGROUND-COLOR: #fff4f0
}
DIV.frmlcrctn#cmtContainer DIV#viewLink {
	BACKGROUND-COLOR: #fff4f0
}
DIV.minrcrctn#cmtContainer DIV#viewLink {
	BACKGROUND-COLOR: #fff4f0
}
DIV.retractionCssStyle#cmtContainer DIV#viewCmtDetail {
	BORDER-BOTTOM-COLOR: #f00; BORDER-TOP-COLOR: #f00
}
DIV.frmlcrctn#cmtContainer DIV#viewCmtDetail {
	BORDER-BOTTOM-COLOR: #f00; BORDER-TOP-COLOR: #f00
}
DIV.minrcrctn#cmtContainer DIV#viewCmtDetail {
	BORDER-BOTTOM-COLOR: #f00; BORDER-TOP-COLOR: #f00
}
DIV#cmtContainer DIV#viewCIStatement {
	PADDING-TOP: 0px
}
DIV#cmtContainer DIV#viewCIStatement DIV.cis {
=09
}
DIV.citation DIV.cis {
=09
}
BLOCKQUOTE DIV.cis {
=09
}
DIV.citation DIV.cis {
	MARGIN-TOP: 5px; PADDING-TOP: 5px
}
DIV#cmtContainer DIV#viewCIStatement DIV.cis {
	FONT-SIZE: 0.9em; PADDING-TOP: 5px
}
DIV#multidetail DIV.cis {
	FONT-SIZE: 0.9em; PADDING-TOP: 5px
}
BLOCKQUOTE DIV.cis {
	PADDING-BOTTOM: 5px; LINE-HEIGHT: 1.5em; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; PADDING-TOP: 5px
}
DIV.dialog .pane.posting {
	BACKGROUND: none transparent scroll repeat 0% 0%
}
DIV.comment DIV.comment {
	BORDER-LEFT: #999 4px solid; BACKGROUND-COLOR: #eee; MARGIN: 10px 0px; =
PADDING-LEFT: 20px
}
DIV.comment .comment H6 {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BACKGROUND: =
none transparent scroll repeat 0% 0%; COLOR: #333; PADDING-TOP: 4px
}
.preview {
	BORDER-BOTTOM: #666 3px solid; BORDER-LEFT: #666 3px solid; WIDTH: =
32em; BACKGROUND: url(../images/dialog_preview_bg.gif) repeat-x 50% top; =
BORDER-TOP: #666 3px solid; BORDER-RIGHT: #666 3px solid
}
.preview DIV.detail A {
	MARGIN: 0px 10px
}
.annotate {
	BORDER-BOTTOM: #666 3px solid; POSITION: relative; BORDER-LEFT: #666 =
3px solid; WIDTH: 52em; BACKGROUND: url(../images/forms_bg.gif) #fff =
repeat-x 50% bottom; BORDER-TOP: #666 3px solid; BORDER-RIGHT: #666 3px =
solid
}
DIV.contextActionDialog {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
DIV.context DIV.tip {
	POSITION: absolute; WIDTH: 39px; BOTTOM: -22px; BACKGROUND: =
url(../images/dialog_pb.gif) no-repeat 0px 0px; HEIGHT: 22px; LEFT: 22px
}
DIV.context DIV.tipu {
	POSITION: absolute; WIDTH: 39px; BACKGROUND: =
url(../images/dialog_pt.gif) no-repeat 0px 0px; HEIGHT: 22px; TOP: =
-22px; LEFT: 22px
}
DIV.context {
	BORDER-BOTTOM: #666 3px solid; POSITION: relative; BORDER-LEFT: #666 =
3px solid; WIDTH: 28em; BACKGROUND: url(../images/forms_bg.gif) #fff =
repeat-x 50% bottom; BORDER-TOP: #666 3px solid; BORDER-RIGHT: #666 3px =
solid
}
DIV.context DIV.contextActionContent {
	BORDER-RIGHT-WIDTH: 0px; MARGIN: 10px; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px
}
DIV.context DIV.contextActionContent H5 {
	PADDING-BOTTOM: 3px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
DIV.context DIV.contextActionContent UL {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 5px
}
DIV.context DIV.contextActionContent LI {
	PADDING-BOTTOM: 0px; MARGIN: 0px 0px 0px 17px; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; PADDING-TOP: 0px
}
DIV.context DIV.contextActionContent INPUT {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; FLOAT: right; PADDING-TOP: 0px
}
DIV.context DIV.contextActionContent FORM {
	PADDING-TOP: 5px
}
DIV.dijitTooltipContainer {
	BORDER-RIGHT-WIDTH: 0px; BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: =
0px; BORDER-LEFT-WIDTH: 0px
}
.loadingCycler {
	Z-INDEX: 99998 !important; OVERFLOW: hidden
}
.loadingCycler IMG {
	Z-INDEX: 99999 !important; OVERFLOW: hidden
}
.comment {
	PADDING-BOTTOM: 10px; PADDING-LEFT: 10px; PADDING-RIGHT: 10px; =
PADDING-TOP: 10px
}
.response FIELDSET {
	BORDER-RIGHT-WIDTH: 0px; BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: =
0px; BORDER-LEFT-WIDTH: 0px
}
.response FIELDSET .reply {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; =
PADDING-BOTTOM: 2px; MARGIN: 10px 0px; PADDING-LEFT: 2px; PADDING-RIGHT: =
2px; DISPLAY: block; BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #ccc 1px =
solid; PADDING-TOP: 2px
}
.posting .title {
	WIDTH: 25em
}
.flag TEXTAREA {
	MARGIN-TOP: 5px; WIDTH: 25em; MARGIN-BOTTOM: 10px; HEIGHT: 8em; =
MARGIN-LEFT: 0px; OVERFLOW: auto
}
.posting TEXTAREA {
	MARGIN-TOP: 5px; WIDTH: 25em; MARGIN-BOTTOM: 10px; HEIGHT: 8em; =
MARGIN-LEFT: 0px; OVERFLOW: auto
}
.response .body {
	WIDTH: 46em; HEIGHT: 12em; OVERFLOW: auto
}
.dojoTooltip {
	Z-INDEX: 10; BORDER-BOTTOM: black 1px solid; POSITION: absolute; =
BORDER-LEFT: black 1px solid; PADDING-BOTTOM: 2px; PADDING-LEFT: 2px; =
PADDING-RIGHT: 2px; DISPLAY: block; BACKGROUND: beige; COLOR: black; =
FONT-SIZE: small; BORDER-TOP: black 1px solid; BORDER-RIGHT: black 1px =
solid; PADDING-TOP: 2px
}
.userinfo {
	Z-INDEX: 10; BORDER-BOTTOM: #333 2px solid; POSITION: absolute; =
TEXT-ALIGN: center; BORDER-LEFT: #333 2px solid; PADDING-BOTTOM: 5px; =
PADDING-LEFT: 5px; WIDTH: 16em; PADDING-RIGHT: 5px; DISPLAY: block; =
FONT-SIZE: 0.9em; BORDER-TOP: #333 2px solid; BORDER-RIGHT: #333 2px =
solid; PADDING-TOP: 5px
}
.userinfo DL {
	LIST-STYLE-TYPE: none; MARGIN: 0px; FONT-SIZE: 1em
}
.userinfo DD {
	LIST-STYLE-TYPE: none; MARGIN: 0px; FONT-SIZE: 1em
}
.userinfo DT {
	LIST-STYLE-TYPE: none; MARGIN: 0px; FONT-SIZE: 1em
}
.userinfo DD {
	FONT-SIZE: 0.9em
}
.pane {
	POSITION: relative; PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: =
5px; PADDING-RIGHT: 5px; PADDING-TOP: 0px
}
.pane.reading {
	HEIGHT: 12em
}
.dialog FIELDSET {
	PADDING-BOTTOM: 0px; BORDER-RIGHT-WIDTH: 0px; MARGIN: 0px; =
PADDING-LEFT: 0px; PADDING-RIGHT: 0px; BORDER-TOP-WIDTH: 0px; =
BORDER-BOTTOM-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px; PADDING-TOP: 0px
}
.dialog .text LABEL {
	DISPLAY: none
}
.dialog LEGEND {
	DISPLAY: none
}
.post LEGEND {
	DISPLAY: none
}
.pane .error {
	BORDER-BOTTOM: #c00 1px solid; BORDER-LEFT: #c00 1px solid; =
PADDING-BOTTOM: 5px; BACKGROUND-COLOR: #fff; FONT-STYLE: normal; =
PADDING-LEFT: 5px; PADDING-RIGHT: 5px; MARGIN-BOTTOM: 5px; BORDER-TOP: =
#c00 1px solid; BORDER-RIGHT: #c00 1px solid; PADDING-TOP: 5px
}
.dialog .response {
	WIDTH: 100%; HEIGHT: 8em; OVERFLOW: auto
}
.dialog .submit INPUT {
	DISPLAY: inline
}
.dialog .submit LABEL {
	DISPLAY: inline
}
.dialog FIELDSET DIV {
	PADDING-BOTTOM: 0px; PADDING-LEFT: 0px; WIDTH: 5em; PADDING-RIGHT: 0px; =
DISPLAY: inline; HEIGHT: 2em; MARGIN-LEFT: 15px; PADDING-TOP: 0px
}
.dialog FIELDSET DIV LABEL {
	WIDTH: 5em; DISPLAY: inline; HEIGHT: 1.12em; VERTICAL-ALIGN: middle
}
.dialog FIELDSET INPUT.radio {
	WIDTH: 1em; DISPLAY: inline; HEIGHT: 1.12em; VERTICAL-ALIGN: middle; =
MARGIN-RIGHT: 4px
}
.dialog FIELDSET INPUT.checkbox {
	WIDTH: 1em; DISPLAY: inline; HEIGHT: 1.12em; VERTICAL-ALIGN: middle; =
MARGIN-RIGHT: 4px
}
DIV.btnwrap {
	MARGIN: 0px; WIDTH: 100%; DISPLAY: block; HEIGHT: auto
}
DIV.btnwrap INPUT {
	PADDING-BOTTOM: 2px; MARGIN: 5px auto; PADDING-LEFT: 2px; =
PADDING-RIGHT: 2px; DISPLAY: block; PADDING-TOP: 2px
}
DIV.multiple {
	POSITION: relative; BACKGROUND-COLOR: #666; WIDTH: 50em
}
.multiple OL {
	PADDING-BOTTOM: 0px; LIST-STYLE-TYPE: none; MARGIN: 0px; PADDING-LEFT: =
0px; PADDING-RIGHT: 0px; FONT-WEIGHT: normal; PADDING-TOP: 0px
}
.multiple LI {
	PADDING-BOTTOM: 0px; LIST-STYLE-TYPE: none; MARGIN: 0px; PADDING-LEFT: =
0px; PADDING-RIGHT: 0px; FONT-WEIGHT: normal; PADDING-TOP: 0px
}
.multiple .detail {
	MARGIN: 0px 10px 10px; FONT-WEIGHT: normal
}
.multiple OL {
	BORDER-BOTTOM: #666 0px solid; POSITION: absolute; BORDER-LEFT: #666 =
1px solid; MARGIN: 0px 10px; WIDTH: 22em; DISPLAY: inline; BORDER-TOP: =
#666 1px solid; TOP: 30px; BORDER-RIGHT: #666 0px solid; LEFT: 0px
}
.multiple LI {
	LIST-STYLE-POSITION: outside; BORDER-BOTTOM: #666 1px solid; =
PADDING-BOTTOM: 5px; TEXT-INDENT: 20px; PADDING-LEFT: 5px; =
PADDING-RIGHT: 5px; BACKGROUND: url(../images/tooltip_addannotation.gif) =
#c8c6c6 no-repeat 5px 0.4em; CURSOR: pointer; BORDER-RIGHT: #666 1px =
solid; PADDING-TOP: 5px
}
.multiple .contentwrap DIV.detail {
	BORDER-RIGHT-WIDTH: 0px; BACKGROUND-COLOR: #fff; MARGIN: 10px 0px 12px; =
BORDER-TOP-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; BORDER-LEFT-WIDTH: 0px
}
.multiple LI DIV.detail {
	BORDER-BOTTOM: #0bf 1px solid; BORDER-LEFT: #0bf 1px solid; =
BACKGROUND-COLOR: #ddd; TEXT-INDENT: 0px; MARGIN: 5px 0px 0px; =
BORDER-TOP: #0bf 1px solid; BORDER-RIGHT: #0bf 1px solid
}
.multiple LI.active {
	BORDER-RIGHT-STYLE: none; BACKGROUND-COLOR: #fff; FONT-WEIGHT: bold
}
.multiple LI.active DIV.detail {
	BACKGROUND-COLOR: #eee
}
.multiple .contentwrap {
	BORDER-BOTTOM: #666 1px solid; BORDER-LEFT: #666 1px; MARGIN: 13px 0px =
0px 22.9em; WIDTH: 23em; DISPLAY: none; BORDER-TOP: #666 1px solid; =
BORDER-RIGHT: #666 1px solid
}
.multiple .active.contentwrap {
	DISPLAY: block
}
.multiple .contentwrap DIV.detail {
	POSITION: absolute; WIDTH: 22.8em; BOTTOM: 24px
}
DIV.multiple OL LI.minrcrctn {
	BACKGROUND-IMAGE: url(../images/icon_correction.gif); =
BACKGROUND-REPEAT: no-repeat; BACKGROUND-POSITION: 5px 0.3em
}
DIV.multiple OL LI.frmlcrctn {
	BACKGROUND-IMAGE: url(../images/icon_correction.gif); =
BACKGROUND-REPEAT: no-repeat; BACKGROUND-POSITION: 5px 0.3em
}
DIV.multiple LI.minrcrctn DIV.detail {
	BORDER-BOTTOM-COLOR: #f00 !important; BORDER-TOP-COLOR: #f00 =
!important; TEXT-INDENT: 0px; BORDER-RIGHT-COLOR: #f00 !important; =
BORDER-LEFT-COLOR: #f00 !important
}
DIV.multiple LI.frmlcrctn DIV.detail {
	BORDER-BOTTOM-COLOR: #f00 !important; BORDER-TOP-COLOR: #f00 =
!important; TEXT-INDENT: 0px; BORDER-RIGHT-COLOR: #f00 !important; =
BORDER-LEFT-COLOR: #f00 !important
}
DIV.multiple LI.minrcrctn {
	COLOR: #f00
}
DIV.multiple LI.frmlcrctn {
	COLOR: #f00
}
DIV.multiple > OL#multilist > LI.active.minrcrctn {
	BACKGROUND-COLOR: #fff4f0
}
DIV.multiple > OL#multilist > LI.active.frmlcrctn {
	BACKGROUND-COLOR: #fff4f0
}
DIV.multiple > DIV#multidetail > DIV.minrcrctn {
	BACKGROUND-COLOR: #fff4f0
}
DIV.multiple > DIV#multidetail > DIV.frmlcrctn {
	BACKGROUND-COLOR: #fff4f0
}
DIV.multiple > DIV#multidetail > DIV.minrcrctn DIV.detail {
	BACKGROUND-COLOR: #fff4f0
}
DIV.multiple > DIV#multidetail > DIV.frmlcrctn DIV.detail {
	BACKGROUND-COLOR: #fff4f0
}
.dialog DT DIV.detail {
	BORDER-BOTTOM-STYLE: none; PADDING-BOTTOM: 0px; BORDER-RIGHT-STYLE: =
none; BACKGROUND-COLOR: transparent; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; BORDER-TOP-STYLE: none; BORDER-LEFT-STYLE: none; PADDING-TOP: 0px
}
.none {
	DISPLAY: none
}
#citation {
	BORDER-BOTTOM: #ccc 2px solid; BORDER-LEFT: #ccc 2px solid; =
PADDING-BOTTOM: 10px; PADDING-LEFT: 10px; PADDING-RIGHT: 10px; HEIGHT: =
12em; OVERFLOW: auto; BORDER-TOP: #ccc 2px solid; BORDER-RIGHT: #ccc 2px =
solid; PADDING-TOP: 10px
}
#citation CITE {
	FONT-STYLE: normal; DISPLAY: block
}
#annotation-box BLOCKQUOTE {
	MARGIN: 0px
}
#annotation-box P {
	MARGIN: 0px
}
DIV.author BLOCKQUOTE {
	BACKGROUND: #fff
}
DIV.author hd {
	BACKGROUND-COLOR: #069
}
DIV.response H3 {
	PADDING-BOTTOM: 0px; MARGIN: 0.15em 0px 0.25em 5px; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; FONT-FAMILY: Verdana, Arial, Helvetica, sans-serif; =
LETTER-SPACING: normal; FONT-SIZE: 1em; FONT-WEIGHT: bold; PADDING-TOP: =
0px
}
DIV.response {
	BORDER-BOTTOM: #ccc 0px solid; BORDER-LEFT: #ccc 0px solid; =
PADDING-BOTTOM: 0px; MARGIN: -2px 0px 0px 15px; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; BORDER-TOP: #ccc 2px solid; BORDER-RIGHT: #ccc 2px =
solid; PADDING-TOP: 0px
}
DIV.response DIV.response {
	BORDER-BOTTOM-STYLE: none; BORDER-RIGHT-STYLE: none; MARGIN: -2px 0px =
0px 20px; BORDER-LEFT-STYLE: none; BORDER-TOP: #ccc 2px solid
}
DIV.response BLOCKQUOTE {
	BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 2px solid; =
PADDING-BOTTOM: 5px; FONT-STYLE: normal; MARGIN: 0px; PADDING-LEFT: =
10px; PADDING-RIGHT: 10px; BACKGROUND: =
url(../images/response_blockquote_bg.gif) #f5f5f6 repeat-x 50% bottom; =
BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #ccc 0px solid; PADDING-TOP: =
5px
}
DIV.response .hd {
	BORDER-LEFT: #ccc 2px solid; PADDING-BOTTOM: 10px; BACKGROUND-COLOR: =
#fff; PADDING-LEFT: 10px; PADDING-RIGHT: 10px; PADDING-TOP: 10px
}
DIV.response .response .hd {
	BACKGROUND: url(../images/response_hd_bg.gif) repeat-x 50% top
}
DIV.response DIV.detail {
	PADDING-LEFT: 5px
}
DIV.response H5 {
	PADDING-BOTTOM: 0px; MARGIN: 20px 5px 5px; PADDING-LEFT: 0px; =
PADDING-RIGHT: 0px; FONT-SIZE: 1.1em; PADDING-TOP: 0px
}
DIV.pane {
	POSITION: relative
}
DIV.pane FIELDSET {
	PADDING-BOTTOM: 0px; MARGIN: 0px; PADDING-LEFT: 0px; PADDING-RIGHT: =
0px; PADDING-TOP: 0px
}
DIV.response DIV.close A {
	POSITION: absolute; FLOAT: none; COLOR: #333; TOP: 20px; RIGHT: 20px
}
DIV.response .pane {
	BORDER-BOTTOM: #ccc 2px solid; BORDER-LEFT: #ccc 2px solid; =
PADDING-BOTTOM: 20px; MARGIN: 0px; PADDING-LEFT: 20px; PADDING-RIGHT: =
20px; BACKGROUND-REPEAT: repeat-x; BACKGROUND-POSITION: 50% top; COLOR: =
#fff; BORDER-TOP: #ccc 2px solid; BORDER-RIGHT: #ccc 0px solid; =
PADDING-TO
