From: Subject: =?Windows-1252?Q?Epidermal_Growth_Factor_Receptor_Expression_Identifies_F?= =?Windows-1252?Q?unctionally_and_Molecularly_Distinct_Tumor-Initiating_Ce?= =?Windows-1252?Q?lls_in_Human_Glioblastoma_Multiforme_and_Is_Required_for?= =?Windows-1252?Q?_Gliomagenesis_=97_Cancer_Res?= Date: Sun, 3 Oct 2010 12:51:18 +0200 MIME-Version: 1.0 Content-Type: multipart/related; type="text/html"; boundary="----=_NextPart_000_0000_01CB62F9.AC22C650" X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.5994 This is a multi-part message in MIME format. ------=_NextPart_000_0000_01CB62F9.AC22C650 Content-Type: text/html; charset="utf-8" Content-Transfer-Encoding: quoted-printable Content-Location: http://cancerres.aacrjournals.org/content/70/19/7500.long =EF=BB=BF Epidermal Growth = Factor Receptor Expression Identifies Functionally and Molecularly = Distinct Tumor-Initiating Cells in Human Glioblastoma Multiforme and Is = Required for Gliomagenesis =E2=80=94 Cancer Res
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Epidermal Growth Factor Receptor Expression = Identifies=20 Functionally and Molecularly Distinct Tumor-Initiating Cells in Human=20 Glioblastoma Multiforme and Is Required for Gliomagenesis

  1. Stefania=20 Mazzoleni= 1,= 5,=20
  2. Letterio=20 S. Politi= 2,=20
  3. Mauro=20 Pala= 6,=20
  4. Manuela=20 Cominelli= 7,=20
  5. Alberto=20 Franzin= 3,=20
  6. Lucia=20 Sergi Sergi= 4,=20
  7. Andrea=20 Falini= 2,=20
  8. Michele=20 De Palma= 4,=20
  9. Alessandro=20 Bulfone= 6,=20
  10. Pietro=20 L. Poliani= 7,=20 and=20
  11. Rossella=20 Galli= 1=20
  1. Authors' Affiliations:1Neural = Stem Cell=20 Biology Unit, Division of Regenerative Medicine, Stem Cells and Gene = Therapy,=20 2Neuroradiology Unit and CERMAC, 3Neurosurgery = Unit, and=20 4Angiogenesis and Tumor Targeting Research Unit, Telethon = Institute=20 for Gene Therapy, Division of Regenerative Medicine, Stem Cells and = Gene=20 Therapy, San Raffaele Scientific Institute; 5Vita-Salute = San=20 Raffaele University, Milan, Italy; 6Bio))flag Ltd., Pula, = Cagliari,=20 Italy; and 7Department of Pathology, University of = Brescia-Spedali=20 Civili of Brescia, Brescia, Italy
  1. Corresponding=20 Author:
    Rossella Galli, Neural Stem Cell Biology Unit, = Division of=20 Regenerative Medicine, Stem Cells and Gene Therapy, San Raffaele = Scientific=20 Institute, Via Olgettina 58, 20132 Milan, Italy. Phone: = 39-02-2643-4626; Fax:=20 39-02-2643-4621; E-mail: galli.rossella{at}hsr.it. =
= Next=20 Section

Abstract

Epidermal growth factor receptor (EGFR) is a known = diagnostic and,=20 although controversial, prognostic marker of human glioblastoma = multiforme=20 (GBM). However, its functional role and biological significance in GBM = remain=20 elusive. Here, we show that multiple GBM cell subpopulations could be = purified=20 from the specimens of patients with GBM and from cancer stem cell (CSC) = lines=20 based on the expression of EGFR and of other putative CSC markers. All = these=20 subpopulations are molecularly and functionally distinct, are = tumorigenic, and=20 need to express EGFR to promote experimental tumorigenesis. Among them,=20 EGFR-expressing tumor-initiating cells (TIC) display the most malignant=20 functional and molecular phenotype. Accordingly, modulation of EGFR = expression=20 by gain-of-function and loss-of-function strategies in GBM CSC lines = enhances=20 and reduces their tumorigenic ability, respectively, suggesting that = EGFR plays=20 a fundamental role in gliomagenesis. These findings open up the = possibility of=20 new therapeutically relevant scenarios, as the presence of functionally=20 heterogeneous EGFRpos and EGFRneg TIC = subpopulations=20 within the same tumor might affect clinical response to treatment. = Cancer Res;=20 70(19); 7500=E2=80=9313. =C2=A92010 AACR.

Previous=20 SectionNext=20 Section

Introduction

In the last 10 years, the identification of putative cancer = stem cells=20 (CSC) in different types of tumors has raised many expectations. CSCs = represent=20 a rare fraction of the tumor bulk, uniquely responsible for tumor = initiation and=20 progression and, as such, are envisioned as elective targets of therapy = (= 1, = 2).=20 The CSC model of tumorigenesis has been convincingly validated in many=20 hemopoietic malignancies (= 3)=20 and in solid tumors such as breast and colon cancers (= 4=E2=80=93= 7).=20 However, in other tumors, CSCs seem to make up the majority of cells = within the=20 tumor bulk and all exhibit tumor-initiating ability, thus being more=20 appropriately called tumor-initiating cells (TIC; refs. = 8=E2=80=9311).=20

This last model might also apply to glioblastoma multiforme = (GBM), as=20 soundly shown by recent studies in cultured CSC lines (12,=20 13).=20 To test whether this concept might hold true in specimens from patients = with=20 primary GBM, we selected the epidermal growth factor receptor (EGFR) as = a=20 prospective GBM CSC marker (14,=20 15).=20 EGFR expression in GBM has both diagnostic and prognostic significance = (16=E2=80=9319)=20 and it is associated with tumor progression (20).=20 Likewise, EGFR gene amplification is positively correlated with = molecularly defined GBM subclasses, characterized by active = proliferation and=20 poor prognosis (21).=20 Valuable preclinical information has also been obtained by modulating = EGFR=20 expression in standard serum-cultured glioma cell lines (22=E2=80=9325)=20 and by using these in vitro cultures for testing different = therapeutic=20 approaches (26).=20

Here, we investigated the biological significance of EGFR = expression=20 in human primary GBM by assessing its functional role in tumor cell = fractions=20 purified from several GBM patient=E2=80=93derived specimens and by = validating it in GBM=20 CSC lines, which reproduce the genotypic and phenotypic characteristics = of GBM=20 more faithfully than standard glioma cell lines (27,=20 28).=20 By this experimental strategy, we identified multiple subpopulations of = GBM=20 TICs, each characterized by a distinct gene signature and tumorigenic = potential,=20 and thus being able to reproduce the diverse pathologic and functional = features=20 of GBM, such as invasive and angiogenic behavior.

Our findings unequivocally associate EGFR expression with = specific=20 biological functions and molecular traits in distinct GBM cell = subpopulations,=20 thus providing relevant preclinical information to be exploited for the=20 development of TIC subpopulation=E2=80=93targeted effective therapies. =

= Previous=20 SectionNext=20 Section

Materials and Methods

Collection and processing of patients' specimens

GBM specimens were collected from patients with histologic = diagnosis=20 of primary GBM (WHO grade 4 glioma) in accordance with the protocol = approved by=20 the institutional review board of San Raffaele Scientific Institute = (01-CSC07).=20 Tumor diagnosis and grading were blind-reviewed by different = pathologists at two=20 distinct institutions. Samples that did not satisfy the inclusion = criteria of=20 primary GBMs (e.g., GBM with oligodendroglioma features) were excluded = from the=20 study. See Supplementary Experimental Procedure for processing methods.=20

Immunohistochemical staining

Four-micrometer sections were cut from paraffin blocks and = were=20 stained with mouse antihuman EGFR, mouse anti-CD31 (all DakoCytomation), = rabbit=20 anti-SOX2 (Abcam), mouse anti-Bmi1, and mouse anti-CD15 (Becton = Dickinson).=20 Sections were then incubated with a secondary antibody (ChemMATE = Envision=20 Rabbit/Mouse, DakoCytomation).

Molecular analysis

Total RNA from GBM tumor specimens and from CSC lines was = extracted=20 using the RNeasy Micro and Mini Kits (Qiagen). cDNA was obtained by = using=20 Superscript RNaseH=E2=88=92 reverse transcriptase = (Invitrogen). All cDNAs=20 were normalized to the =CE=B2-actin reverse transcription-PCR (RT-PCR) = product.=20 Standard RT-PCR for EGFRvIII was done as described in ref. 29.=20

Fluorescence-activated cell sorting

Following enzymatic and mechanical dissociation, cells from = primary=20 GBM tissues and from experimental xenografts were stained with rabbit = antihuman=20 EGFR (clone EGFR.1, AbD Serotec), rat antihuman CD34/CD45/CD11b (Becton=20 Dickinson), antihuman AC133 (Miltenyi), mouse anti-CD15 (clone MMA, = Becton=20 Dickinson), and antihuman pan-HLA (Becton Dickinson). Cells were sorted = on a=20 Becton Dickinson FACSVantage SE FACSDiVa. The purity of each cell = fraction=20 ranged between 90% and 99%. Fluorescence-activated cell sorting (FACS) = analysis=20 of CSC lines was done as described above, without enzymatic digestion.=20

Culture propagation, population analysis, and cloning

Established GBM CSCs were cultured as previously described = (27).=20

Microarray-based gene expression profiling

See Supplementary Experimental Procedure for = details.

Evaluation of CSC tumorigenicity by orthotopic implantation

Tumorigenicity was determined by injecting all the distinct = cell=20 preparations orthotopically in nu/nu mice (27).=20 Magnetic resonance imaging (MRI) analysis was performed as described in=20 Supplementary Experimental Procedure.

Immunofluorescence analysis was performed on cryostat = sections using=20 mouse monoclonal antihuman EGFR (1:100; Calbiochem). Details can be = found in=20 Supplementary Data.

Western blotting

Lysates from GBM tissues and CSCs were immunoblotted = according to=20 standard protocols. The primary antibodies used were rabbit anti-EGFR, = rabbit=20 anti=E2=80=93phosphorylated EGFR (Tyr1068; Cell Signaling), = rabbit anti-ErbB2=20 (Calbiochem) and rabbit anti-Erbb3 (Santa Cruz Biotechnology), rabbit=20 anti=E2=80=93phosphorylated mitogen-activated protein kinase (MAPK; Cell = Signaling), and=20 rabbit anti=E2=80=93phosphorylated Akt (Ser473; Cell = Signaling). As a loading=20 control, a mouse anti=E2=80=93=CE=B2-tubulin antibody was used = (Sigma-Aldrich).

Lentiviral vector generation, production, and cell infection

Human wild-type (wt) EGFR cDNA (Upstate) was cloned into = the=20 monocistronic transfer lentiviral vector pCCL.sin.cPPT.PGK.GFP.WPRE11. = CSCs were=20 transduced with 1 =C3=97 107 transduction units/mL of = lentiviral vectors=20 for 16 hours.

Lentiviral vectors coding for shRNAs targeted against the = human EGFR=20 were purchased from Sigma (Mission RNAi). Infection of CSCs was = performed=20 according to the manufacturer's instructions.

Immunofluorescence and fluorescence in situ hybridization =

Immunofluorescence was performed as described in ref. 27,=20 using mouse antihuman EGFR (Calbiochem) and rabbit polyclonal anti-GFP=20 (Molecular Probes). Fluorescence in situ hybridization (FISH) = was=20 performed using the kit from Dako, according to the manufacturer's = instructions.=20 Filamentous actin (F-actin) was stained by TRITC-conjugated phalloidin=20 (Chemicon).

Pharmacologic EGFR inhibition in CSCs

AG1478 (Calbiochem) was used at the indicated final = concentrations in=20 complete medium. DMSO was used at the same concentration as the vehicle = control.=20 Cell proliferation was measured by an MTT incorporation assay. =

Invasion assays

Invasion assays were performed in Matrigel-coated = 8-=CE=BCm-pore Transwell=20 chambers (Corning Costar). EGFRpos and EGFRneg = CSCs were=20 seeded in sister cultures on the upper side of the chambers in complete = medium=20 and allowed to migrate for 5 and 7 days, respectively, with or without = AG1478.=20 Noninvaded cells on the upper side of the filters were or were not = scraped off,=20 and those that migrated to the lower side were fixed and stained by = using=20 DiffQuick (Dade Behring). The extent of cell migration was normalized to = the=20 total number of viable cells in the corresponding unscraped culture.=20

Previous=20 SectionNext=20 Section

Results

EGFR expression identifies distinct cell subpopulations within human = primary=20 GBM and GBM CSC lines

Several GBM surgical specimens were tested for EGFR mRNA = expression=20 by both microarrays and semiquantitative RT-PCR and all were shown to = express=20 the transcript (Tab= le=20 1). However, immunohistochemistry on corresponding paraffin-embedded = sections showed that, in agreement with the literature, only 50% of = specimens=20 were characterized by diffuse EGFR staining, which was particularly = intense in=20 small-sized round tumor cells (EGFRpos tumor samples; Fig= . 1A,=20 Supplementary Fig. S1). Accordingly, the other specimens displayed = negligible=20 EGFR expression that was restricted mostly to tumor-infiltrating = inflammatory=20 cells (EGFRneg tumor samples; Fig= .=20 1A). Western blotting (WB) analysis confirmed the = immunohistochemistry=20 findings (Fig= .=20 1B). Hence, EGFR protein distribution identifies two main distinct = subtypes=20 of human GBM (16).=20

View this table:
Table 1.=20

EGFR expression in GBM patients' tumor=20 specimens

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Figure 1.=20

EGFR expression identifies distinct = cell=20 subpopulations within primary human GBM. A, EGFRpos GBM = tumors show=20 diffuse and intense reactivity for EGFR in small and round neoplastic = cells=20 (T1224). EGFRneg GBM specimens show EGFR staining restricted = to=20 tumor-infiltrating inflammatory cells (black arrow; T0109). = Magnification, =C3=97400=20 and =C3=97800. B, WB confirms EGFR protein levels in GBM patients' = specimens. C, FISH=20 analysis indicates that the EGFR gene is amplified only in=20 EGFRpos tumors. D, the mutant EGFRvIII can be detected in a = subset of=20 EGFRpos tumors by RT-PCR. E, EGFRpos tumors = contain=20 modestly proliferating vessels (arrows), whereas EGFRneg = tumors show=20 an enlarged and proliferating vasculature (arrows). H&E and CD31 = staining;=20 magnification, =C3=97200 and =C3=97400. F, EGFRpos and = EGFRneg=20 tumors contain similar numbers of Sox2 and Bmi1pos cells.=20 CD15pos proper tumor cells can be detected only in = EGFRpos=20 (arrow at bottom right, CD15pos monocytes within a vessel).=20 Magnification, =C3=97400. G, in EGFRpos tumors, many = EGFRpos=20 cells are proper tumor cells, whereas in EGFRneg tumors the = few=20 EGFRpos cells are stromal/inflammatory cells. H, the=20 EGFRpos fractions from EGFRpos specimens contain=20 AC133pos and CD15pos cells (top middle), which are = either=20 tumor cells or stromal cells. On the contrary, EGFRneg = fractions (top=20 right) and EGFRneg GBM tumors (bottom) contain only stromal=20 AC133pos and CD15pos cells.

By FISH, a subset of EGFRpos tumor specimens = showed=20 EGFR gene amplification and expression of mutant EGFRvIII that, = in some=20 patients, correlated with EGFR protein levels (Fig= . 1C and=20 D; Tab= le=20 1). Interestingly, EGFRpos tumors contained small and = thin=20 CD31-positive vessels (Fig= .=20 1E), whereas EGFRneg tumors were characterized by an = actively=20 proliferating vasculature [mean vessel area (=C2=B1SEM), 853.1 =C2=B1 = 214.1 and 2,500.2 =C2=B1=20 615.1 =CE=BCm2 in EGFRpos and EGFRneg = GBMs,=20 respectively; P > 0.05, Student's t test; = n =3D 7;=20 Fig= .=20 1E]. All specimens expressed similar levels of stem = cell=E2=80=93associated=20 proteins, such as Sox2 and BMI1 (Fig= .=20 1F), and of glial fibrillary acidic protein (GFAP; data not shown). =

Interestingly, whereas immunohistochemistry indicated that = EGFR was=20 expressed in the totality of GBM cells, flow cytometry suggested that = EGFR=20 expression could be retrieved only in a smaller proportion of cells. Of = note,=20 within this EGFRpos cell fraction, a conspicuous subset of = cells=20 colabeled with the endothelial marker CD34 and the hematopoietic markers = CD45=20 and CD11b (Fig= .=20 1G), indicating that they were stromal cells. The CSC marker CD133 = (30)=20 was more expressed in proper tumor cells than in stromal cells in=20 EGFRpos tumors, whereas the opposite was detected in=20 EGFRneg tumors. On the contrary, CD15 was always more = expressed in=20 the stromal cell compartment in both EGFRpos and = EGFRneg=20 tumors (refs. 31,=20 32;=20 Supplementary Table S1).

Thus, the frequency of EGFRpos proper tumor = cells in GBM,=20 calculated after the exclusion of stromal cells, ranged from 0% to 35% = of the=20 total tumor cell number (Tab= le=20 1). Of note, some EGFRpos proper tumor cells also labeled = with=20 AC133 and/or CD15 markers, whereas EGFRneg cells did not (Fig= . 1F and=20 H; Supplementary Table S2).

To formally show that EGFR might also play a role in the = biology of=20 GBM CSCs in vitro, we tested 13 GBM CSC lines previously = validated for=20 being bona fide CSC lines (Supplementary Fig. S2). All CSC lines = expressed EGFR=20 mRNA, whereas only 5 of 13 CSC lines also expressed the EGFR protein (Tab= le=20 2). Among the EGFRpos CSC lines, only 6% to 70% of cells=20 expressed the receptor, suggesting that multiple GBM subpopulations = could also=20 be identified based on EGFR expression in CSC lines.

View this table:
Table 2.=20

Characterization of EGFRpos = and=20 EGFRneg GBM CSC lines for their expression of the putative = CSC=20 enrichment markers AC133 and CD15

Interestingly, many EGFRpos and = EGFRneg CSC=20 lines contained AC133pos and CD15pos cells (Tab= le 2;=20 Supplementary Table S3). However, one of five EGFRpos CSC = lines and=20 four of eight EGFRneg CSC lines did not contain any=20 AC133pos and CD15pos cells at all.

EGFR expression identifies GBM subpopulations with a distinctive = gene=20 signature

To molecularly analyze the above-described GBM = subpopulations, we=20 purified them from (a) EGFRpos single patients' = tumor=20 specimens, (b) EGFRneg specimens, and (c)=20 EGFRpos CSC lines.

EGFRpos patient samples T1224, T0131, T0222, and = T0321=20 contained (a) EGFRpos/AC133neg cells = (region R1),=20 (b) EGFRpos/AC133pos cells (R2), and = (c)=20 EGFRneg/AC133neg (R3) cells. Two of them also = contained a=20 small proportion of EGFRpos/CD15pos cells (Fig= . 2A and=20 B; Supplementary Table S2). Stromal cells were excluded from = sorting.

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Figure 2.=20

EGFR expression identifies GBM = subpopulations with=20 distinctive gene signatures and different growth characteristics in=20 vitro. A and B, EGFRpos/AC133neg (R1),=20 EGFRpos/AC133pos (R2), and=20 EGFRneg/AC133neg (R3) cells were isolated from=20 EGFRpos tumor specimens by FACS (IgGs, isotype controls). C, = global=20 gene expression data in R1, R2, and R3 fractions purified from patients' = specimens (left). The expression of a selection of coregulated genes is=20 highlighted in the heatmaps (right). D, limiting dilution clonogenic = assay of=20 R1, R2, and R3 fractions immediately after sorting. E, different CSC = lines=20 comprise variable amounts of EGFRpos cells. Magnification, = =C3=97200=20 (insets, =C3=97600). F, some EGFRpos CSCs display = EGFR gene=20 amplification. G, different CSC subpopulations could be isolated by FACS = from=20 EGFRpos CSC lines based on their EGFR expression (region R4,=20 EGFRhigh CSCs; R5, EGFRlow CSCs; R6, = EGFRneg=20 CSCs; L0627). H, WB analysis shows that EGFR is activated in = EGFRhigh=20 and EGFRlow CSCs but is absent in EGFRneg CSCs. I, = global=20 gene expression data of R4, R5, and R6 GBM fractions purified from=20 EGFRpos CSC lines. Semiquantitative RT-PCR validation of = DEGs. J,=20 clonogenic assay and long-term growth curves of R4, R5, and R6 CSCs = immediately=20 after sorting and after in vitro culture. K, EGFR expression in = CSC=20 fractions immediately after sorting (top) and on in vitro = culture=20 (bottom; EGFR, green; magnification, =C3=97200).

Immediately after cell sorting and purity assessment, = without any=20 in vitro manipulation, the different GBM fractions were = subjected to=20 transcriptional profiling by whole-transcript microarrays (Fig= .=20 2C). EGFRpos cell subpopulations displayed a specific = molecular=20 signature clearly distinguishable from that of double-negative cells = (R3). Of=20 note, a subset of 163 genes was differentially expressed between=20 EGFRpos and EGFRneg cell subpopulations, even when = isolated from distinct patient specimens, suggesting that EGFR = expression in GBM=20 might affect common molecular determinants (Supplementary List S1). In = addition=20 to EGFR, the differentially expressed genes (DEG) upregulated in=20 EGFRpos versus EGFRneg GBM fractions were related = to=20 biological processes such as tumor invasion and progression [e.g., = periostin=20 (POSTN), LMO3, and AQP4]. Conversely, genes=20 overexpressed in EGFRneg GBM fractions were mostly involved = in the=20 interaction of tumor cells with the microenvironment [e.g., PDGFRB, = Notch3,=20 PTPRK, lumican (LUM), and endothelin receptor A = (EDNRA)].=20 Thus, GBMs contain molecularly distinct subpopulations of tumor cells. =

When the same freshly isolated cell fractions were cultured = in=20 vitro by the NeuroSphere Assay under limiting dilution conditions = (33),=20 EGFRpos cell fractions gave rise to a higher frequency of = secondary=20 neurospheres than EGFRneg cells (Fig= .=20 2D).

Cell subpopulations expressing different levels of EGFR = were also=20 purified from EGFRpos CSC lines (Fig= .=20 2E=E2=80=93G). Immediately after sorting, EGFRhigh (R4),=20 EGFRlow (R5), and EGFRneg (R6) CSC subpopulations = were=20 validated by WB (Fig= . 2H)=20 and subjected to transcriptional profiling, which indicated that each=20 subpopulation was molecularly distinct from the others (Fig= . 2I;=20 Supplementary List S2). Of note, genes upregulated in = EGFRhigh and=20 EGFRlow versus EGFRneg fractions were also highly=20 expressed in the most malignant molecular subclasses of primary GBM = (i.e., the=20 proliferative and mesenchymal subtypes, as identified in refs. 21=20 and 34;=20 Supplementary Fig. S3). Likewise, genes downregulated in = EGFRpos=20 fractions were also downregulated in the same malignant molecular = subtypes.

As shown by clonogenic assays performed under limiting = dilution=20 conditions and by long-term population analysis, EGFRlow and=20 EGFRneg cell fractions displayed the highest proliferative = capacity,=20 whereas EGFRhigh cell subpopulations showed a transiently = reduced=20 growth ability that required a few subculturing passages to be = completely=20 restored to the presorting levels (Fig= .=20 2J). All subpopulations contained the same percentage of Bmi1- and=20 Sox2-expressing cells, which was not affected by cell differentiation=20 (Supplementary Fig. S4).

When tested immediately after sorting, EGFRhigh = and=20 EGFRlow fractions contained 100% of EGFRpos cells, = whereas=20 EGFRneg fractions were devoid of any EGFRpos cells = (Fig= .=20 2K). However, after several subculturing passages, each cell = fraction=20 regenerated the same frequency of EGFRpos and = EGFRneg=20 cells found in the parental bulk population, recapitulating the original = cellular heterogeneity (Fig= .=20 2K).

EGFR expression confers increased tumorigenic capacity to GBM = cells

To assess whether EGFR expression might affect the = tumorigenic=20 ability of GBM cells, R1, R2, and R3 fractions, purified from = EGFRpos=20 GBM specimens, were transplanted into the brain of nu/nu mice, = without=20 any prior in vitro manipulation (Fig= . 3A;=20 Supplementary Fig. S1). MRI-based volumetric analysis showed that=20 EGFRpos cells were characterized by enhanced tumorigenic = capacity (Fig= .=20 3A). However, EGFRneg/AC133neg cells were also = capable=20 of initiating tumors, although these developed significantly more slowly = than=20 those generated by EGFRpos GBM cells (Kaplan-Meier analysis; = Fig= .=20 3B).

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Figure 3.=20

EGFR expression identifies TICs with = different=20 tumorigenic capacities. A, EGFRpos cell=E2=80=93derived = tumors, independently=20 of the coexpression of AC133, were significantly larger than tumors = induced from=20 EGFRnegAC133neg cells. P < 0.05, = one-way ANOVA=20 with Bonferroni test (n =3D 5, six independent experiments). B, = Kaplan-Meier survival curves (P < 0.01). C, EGFR expression = detected=20 by an antihuman EGFR is retrieved in all xenografts, including=20 EGFRnegAC133neg cell=E2=80=93derived tumors (EGFR, = green).=20 EGFRpos cell=E2=80=93derived tumors are characterized by an = infiltrative=20 pattern of growth. Tumor cells disperse within the gray matter, avoiding = the=20 corpus callosum (white arrow). EGFRnegAC133neg = cells give=20 rise to expanding tumors, mostly confined within the injection site, = which=20 migrate and invade along white matter tracts (white arrowhead). D, WB = confirms=20 EGFR protein expression in all xenografts. E, the same distinct=20 EGFRpos and EGFRneg subpopulations as detected in = the=20 parental tumor were retrieved in the corresponding xenografts by flow = cytometry.=20 CD15pos cells (F) were not retrieved in any xenografts by=20 immunohistochemistry. G, a similar frequency of Sox2 and Bmi1 = immunoreactive=20 cells could be retrieved in R1- and R3-derived xenografts. H, global = gene=20 expression data of R1- and R3-derived xenografts. I to J, EGFR = expression=20 confers enhanced tumorigenic potential to EGFRpos fractions = from=20 EGFRpos CSC lines (one-way ANOVA with Bonferroni test, = n =3D=20 5, three independent experiments). P < 0.01, Kaplan-Meier = survival=20 curves. K, EGFR expression was retrieved in all xenografts, whereas = ErbB2 was=20 highly expressed only in EGFRhigh CSC=E2=80=93derived tumors = (WB).

Notably, EGFR expression was retrieved not only in = xenografts derived=20 from EGFRpos cells but also in those derived from = EGFRneg=20 fractions (Fig= .=20 3C=E2=80=93E). Importantly, the latter xenografts never contained = either=20 AC133pos cells (Fig= . 3E)=20 or CD15pos cells (Fig= . 3F,=20 Supplementary Fig. S1). Thus, as recently shown for other stem cell = markers in=20 melanoma (35),=20 EGFR expression in GBM TICs is dynamically regulated, given that=20 EGFRneg cells can become EGFRpos on = transplantation. Most=20 importantly, EGFR expression positively correlates with gliomagenesis. =

Although R1-, R2-, and R3-derived xenografts showed the = same=20 expression of SOX2 and Bmi1 (Fig= . 3G=20 and data not shown), they were molecularly different (Fig= .=20 3H). By microarray-based analysis, proinvasive genes were = overexpressed in=20 EGFRpos cell=E2=80=93derived xenografts, whereas genes = involved in=20 angiogenesis were upregulated in EGFRneg cell=E2=80=93derived = xenografts=20 (Supplementary Fig. S5).

In agreement with these latter findings and with molecular = data=20 obtained on EGFRpos and EGFRneg subpopulations (Fig= .=20 2C), xenografts from EGFRpos cells were shown to be = highly=20 invasive, with cells spreading preferentially along the gray matter = while=20 avoiding white matter fiber tracts (Fig= . 3C and=20 G; Supplementary Figs. S1 and S6). On the contrary, tumors formed by = EGFRneg cells showed well-demarcated boundaries, with cells = migrating=20 preferentially along white matter tracts (Fig= . 3C and=20 G; Supplementary Figs. S1 and S6). Accordingly, EGFRpos=20 cell=E2=80=93induced xenografts were characterized by thin, regularly = shaped blood=20 vessels and a low proliferative index, whereas tumors from = EGFRneg=20 cells contained enlarged and proliferating vessels and a higher = frequency of=20 mitotic cells (Supplementary Figs. S1 and S6).

Thus, EGFR expression confers different invasive, = angiogenic, and=20 tumorigenic abilities to GBM cells in vivo.

Notably, the same functional differences observed between = xenografts=20 generated by EGFRpos and EGFRneg TICs purified = from GBM=20 specimens could also be detected in xenografts generated by in = vitro=20 expanded EGFRpos and EGFRneg CSC lines = (Supplementary Fig.=20 S2; ref. 36).=20

Because EGFR expression in EGFRneg = cell=E2=80=93derived xenografts=20 might be due to contaminating EGFRpos tumor cells, we = purified=20 EGFRneg tumor cells from EGFRneg specimens that = were=20 totally devoid of EGFRpos, AC133pos, and=20 CD15pos proper tumor cells (Fig= . 1A, G,=20 and H; Supplementary Table S1). FACS-purified=20 EGFRneg/AC133neg cells generated xenografts that = expressed=20 EGFR but were negative for AC133 and CD15, thus confirming that EGFR = might=20 indeed play a prominent role in glioma formation (Supplementary Fig. = S7).=20 Notably, both in vivo limiting dilution analysis and serial=20 transplantation proved that all GBM subpopulations identified based on = EGFR=20 expression could be considered bona fide TICs (Supplementary Fig. S8). =

The tumorigenic potential of EGFRhigh, = EGFRlow,=20 and EGFRneg cells purified from different EGFRpos = CSC=20 lines (Fig= s. 2G=20 and 3I = and J=20 and Supplementary Fig. S9 for L0627 and Supplementary Fig. S9 for L0605) = was=20 also tested. As observed for xenografts obtained from GBM = specimen=E2=80=93derived=20 subpopulations (Fig= .=20 3A=E2=80=93E), tumors derived from EGFRhigh and = EGFRlow CSCs=20 were larger than those generated from EGFRneg cell fractions, = which,=20 again, displayed tumor-initiating ability (Fig= . 3I and=20 J). Again, EGFRneg CSC fraction=E2=80=93derived tumors = expressed EGFR,=20 confirming that EGFR upregulation occurred during experimental = tumorigenesis (Fig= .=20 3K). ErbB2 expression was detected only in EGFRhigh = cell=E2=80=93derived=20 tumors, suggesting that the xenografts were molecularly heterogeneous = (Fig= .=20 3K).

To understand the relationship between the expression of = EGFR and=20 that of the CSC markers CD15 and AC133 in terms of tumor-initiating = potential,=20 EGFRposCD15neg (R4), = EGFRposCD15pos=20 (R5), EGFRnegCD15pos (R6), and=20 EGFRnegCD15neg (R7) as well as=20 EGFRposCD15negAC133neg (R1) and=20 EGFRnegCD15negAC133neg (R3) fractions = were=20 purified from L0627 and L0605 (Supplementary Fig. S9). Interestingly,=20 EGFRpos subpopulations, independent of the coexpression of = AC133=20 and/or CD15, generated tumors that developed much earlier and grew = larger than=20 those generated by EGFRneg fractions. Consistent with = previous=20 findings, CSC lines/subpopulations negative for EGFR, AC133, and CD15 = generated=20 xenografts that did not express either of the two markers in = vivo but=20 did express the EGFR (Supplementary Figs. S2 and S9).

Thus, all of our GBM CSC lines, taken as a whole or = fractionated into=20 subpopulations and independent of their basal EGFR expression, gave rise = to=20 xenografts that upregulated the EGFR significantly (Fig. 3K; = Supplementary Figs.=20 S2 and S9). To understand the mechanism underlying EGFR expression in=20 EGFRneg cell=E2=80=93derived xenografts, we hypothesized that = because=20 EGFRneg GBM cells retain EGFR transcript expression (Tab= les 1=20 and 2; Fig= . 2C and=20 I), in vivo EGFR expression might be regulated=20 posttranscriptionally. Indeed, when we simulated in vitro the=20 mitogen-reduced adult brain microenvironment by starvation, = EGFRneg=20 CSCs efficiently reexpressed the EGFR protein (Supplementary Fig. S2).=20 Accordingly, when the same CSCs were reexposed to standard mitogen=20 concentrations, they downregulated EGFR expression, as may occur in=20 vivo during tumor progression when tumor-infiltrating stromal cells = secrete=20 mitogens in a paracrine fashion (37).=20

Enforced EGFR expression by gene transfer increases the malignant = behavior=20 of CSCs in vitro and in vivo

Lentiviral vector=E2=80=93mediated EGFR expression in three = distinct=20 EGFRneg CSC lines allowed the expression of a fully = functional=20 receptor (Fig= .=20 4A). Of note, the majority of EGFR-transduced cells acquired a = fibroblastoid=20 shape and developed stress fibers and actin-driven lamellipodia, all = known=20 markers of epithelial-mesenchymal transition and enhanced malignancy (Fig= . 4B;=20 Supplementary Table S4). Consistently, the average cell size was = increased in=20 most EGFR-transduced cells (Fig= . 4C;=20 Supplementary Table S4).

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  • Downl= oad as=20 PowerPoint Slide
Figure 4.=20

Overexpression of EGFR enhances the = tumorigenic=20 potential of CSCs. A, lentiviral-mediated EGFR gene transfer = resulted=20 in the expression of a functional EGFR, as indicated by phosphorylation = at=20 residue Tyr1068 and by the hyperactivation of downstream = pathways. B,=20 enforced EGFR expression in EGFRneg CSCs (EGFR, red) = correlates with=20 changes in cell morphology, which are absent in GFP-transduced CSCs = (GFP,=20 green). F-actin staining (phalloidin, red). Magnifications: top, = =C3=97400; insets,=20 =C3=971,000; bottom, =C3=971,000. C, EGFR overexpression in = EGFRneg CSCs=20 induces a significant increase in their size. FSC-A and SSC-A. D, global = gene=20 expression data in na=C3=AFve, mock-transduced, and EGFR-transduced CSC = lines=20 identify a subset of DEGs that are coregulated in different = patient-derived CSC=20 lines and modulated by enforced EGFR expression (EGFR gene = signature).=20 E, semiquantitative RT-PCR validation of DEGs. F, ectopic expression of = EGFR in=20 EGFRneg CSC lines increases their tumorigenic potential and = reduces=20 mouse survival (Student's t test, n =3D 5, three = independent=20 experiments).

Gene expression profiling of na=C3=AFve, mock, and = EGFR-overexpressing CSC=20 lines identified a common subset of DEGs that was altered in both L0104 = and=20 L0125 CSC lines, suggesting that enforced EGFR expression affected the = same=20 molecular determinants in different patient-derived CSC lines (Fig= . 4D and=20 E). In addition to EGFR, LIF, and ITGA3, genes = previously=20 shown to be also highly expressed in FACS-purified EGFRpos = CSC=20 fractions (Fig= . 2I;=20 Supplementary List S2), proinvasive genes such as CXCL10 and=20 ICAM1 (38,=20 39)=20 were upregulated in EGFR-transduced CSCs (Fig= .=20 4D=E2=80=93E) and also overexpressed in the proliferative and = mesenchymal subtypes=20 of human GBM (ref. 2I;=20 Supplementary Fig. S3). Accordingly, tumor suppressors such as = SPARCL1=20 (Fig= .=20 2I), ERBB4, ASCL1, and EDNRB, which were = downregulated in=20 EGFR-transduced CSCs, were also downregulated in GBM malignant subtypes. =

Notably, all EGFR-transduced CSC lines formed experimental = tumors=20 that were larger than those induced by the corresponding mock lines (Fig= .=20 4F).

Genetic loss of function of EGFR reduces the malignancy of CSCs = in=20 vitro and in vivo

To silence EGFR expression stably, we exploited line L0201 = and line=20 L0627, which express high levels of EGFR in 70% and 30% of the cells,=20 respectively (Tab= le=20 2). The shRNA clones 204 and 206 induced EGFR protein knockdown with = an=20 efficiency of 60% and 70% in lines L0201 (Fig= . 5A)=20 and L0627 (data not shown), respectively. WB analysis confirmed the flow = cytometry results and indicated an inhibition of downstream signaling = pathways=20 on EGFR silencing (Fig= .=20 5B).

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  • Downl= oad as=20 PowerPoint Slide
Figure 5.=20

Inhibition of EGFR expression and = activity=20 negatively affects the malignant behavior of CSCs. A and B, flow = cytometry and=20 WB of L0201 shRNA 204 and shRNA 206 CSC subclones indicate efficient = knockdown=20 of EGFR protein and downregulation of MAPK and Akt downstream pathways. = C and D,=20 EGFR knockdown induces CSC differentiation. Tuj1, red; GFAP, green.=20 Magnifications: top, =C3=97200; bottom, =C3=97400. *, P > = 0.05, Student's=20 t test. E, EGFR silencing in CSCs significantly decreases their = tumorigenicity (one-way ANOVA with Bonferroni test, n =3D 4, = two=20 independent experiments). P < 0.01, Kaplan-Meier survival = curves. F,=20 AG1478 treatment inhibits the phosphorylation status of EGFR and Akt in = CSCs=20 (WB). G, EGFRpos CSC lines respond to treatment by decreasing = their=20 proliferation/survival, whereas EGFRneg CSC do not. *, = P=20 > 0.05; **, P > 0.01, Student's t test. H and I, = EGFR=20 inhibition by AG1478 negatively affects the invasive ability of CSCs. = **,=20 P > 0.01, Student's t test. J, CSC responsiveness = to TKI is=20 independent of PTEN expression (WB).

EGFR-knockdown CSCs displayed morphologic changes in = vitro,=20 with neurospheres appearing as well-differentiated cells (Fig= .=20 5C). Consistently, the frequency of differentiated cells detected in = shRNA-transduced CSCs increased under both proliferative (Fig= . 5D)=20 and differentiative culture conditions (data not shown).

Of note, tumors generated from EGFR-knockdown CSCs = developed more=20 slowly and were significantly smaller than those generated by the CTRL = subclone,=20 thus resulting in the increased survival of EGFR-knockdown = CSC=E2=80=93transplanted mice=20 (Fig= .=20 5E).

EGFRpos and EGFRneg CSC lines differentially = respond=20 to pharmacologic inhibition of EGFR

To pharmacologically inhibit EGFR activity, we exploited = the EGFR=20 tyrosine kinase inhibitor (TKI) AG1478. WB analysis of two different=20 EGFRpos CSCs that were exposed to AG1478 for 2, 5, and 16 = hours=20 showed effective inhibition of EGFR autophosphorylation and a = significant=20 decrease in AKT phosphorylation, a hallmark of drug sensitivity (Fig= .=20 5F).

Pharmacologic treatment of EGFRpos CSC lines = with AG1478=20 for up to 72 hours in vitro resulted in a significant decrease = in CSC=20 survival/proliferation (Fig= .=20 5G). Relevantly, the survival and proliferation of = EGFRneg CSC=20 lines were not affected by AG1478, even when the same lines were = transduced with=20 EGFR-coding vectors before treatment, suggesting that = EGFR-overexpressing=20 EGFRneg CSC lines did not become addicted to EGFR (data not = shown).=20 Likewise, exposure of EGFRpos CSCs to AG1478 strongly reduced = their=20 invasive capacity as compared with controls (Fig= . 5H and=20 I). Again, the invasive ability of EGFRneg CSC lines was = not=20 affected by AG1478 exposure.

Interestingly, all the EGFRpos CSC lines = responded to=20 pharmacologic treatment independently of the expression of PTEN, a = putative=20 molecular determinant of TKI responsiveness in patients with GBM (Fig= . 5J;=20 ref. 40).=20

Thus, EGFRpos CSCs are responsive to = pharmacologic=20 inhibition by TKIs, whereas EGFRneg CSCs are not. =

Previous=20 SectionNext=20 Section

Discussion

EGFR expression contributes to GBM heterogeneity

GBM is characterized by a high degree of heterogeneity, = which is=20 efficaciously reflected in the definition =E2=80=9Cmultiforme.=E2=80=9D = However, the intrinsic=20 heterogeneity of GBM might refer not only to the well-known molecular = diversity=20 of GBM subtypes but also to the existence of genetically and = functionally=20 distinct cohorts of TICs within the same tumor.

In line with recent reports that provide new = interpretations of the=20 process of tumorigenesis in mouse hematopoietic malignancies (= 8),=20 mouse colon cancer (41),=20 human melanoma (10,=20 11),=20 and brain tumors (42),=20 our findings indicate that the initiation and progression of GBM may not = depend=20 on the presence of a single and rare fraction of CSCs, as proposed by = the=20 hierarchical model of tumor development (43).=20 On the contrary, our data propose a different biological context, in = which all=20 GBM cells can be considered TICs with different degrees of stemness as = proven by=20 their different tumorigenic ability and by their distinct phenotypic and = molecular features (44).=20 In fact, all of our GBM-derived TIC subpopulations satisfy the = requirements for=20 bona fide CSCs: They self-renew in vitro and in vivo = and give=20 rise to experimental tumors that recapitulate the phenotypic traits of = the tumor=20 of origin under limiting dilution conditions and on serial = transplantation.

Based on these observations, GBM tissue heterogeneity might = be=20 explained by the presence of a hierarchy of distinct TICs, as recently = suggested=20 for in vitro generated GBM CSC lines (13).=20 These distinct TICs distribute along a gradient of tumorigenic potential = that is=20 strictly associated with EGFR expression.

Thus, our study provides conclusive evidence that the = majority of GBM=20 cells have tumorigenic capacity and that the hierarchical model of = tumorigenesis=20 might not fully apply to GBM.

Mechanisms underlying EGFR-dependent GBM heterogeneity

Tumor cells might modify their phenotype during tumor = progression,=20 with multiple cohorts of TICs being generated (45).=20 In this study, we have shown that EGFR expression contributes to GBM = cell=20 diversity by generating different TIC populations. As to the mechanisms = involved=20 in the generation of EGFR-dependent phenotypic heterogeneity in GBM, the = contribution of tumor microenvironment needs to be taken into = consideration.=20 Paracrine EGFR ligands, such as epidermal growth factor, are released by = tumor=20 stromal cells that, in this way, contribute to the activation of the = EGFR=20 signaling in both proper tumor cells and endothelial cells (37).=20 When stroma-produced epidermal growth factor is secreted, EGFR protein=20 downregulation occurs in target cells by receptor internalization, = followed by=20 lysosomal degradation.

In line with this view, it is tempting to speculate that = the=20 EGFRneg cell component detected in EGFRpos GBM = might be=20 generated from GBM cells that were originally EGFRpos through = receptor downregulation induced by excessive ligand stimulation during = tumor=20 progression, as we simulated in vitro by modulation of mitogen=20 concentration. In addition, termination of EGFR signaling in = EGFRneg=20 GBM cells might lead to the acquisition of compensatory additional = genetic=20 events, which might account for the molecular differences detected = between=20 EGFRpos and EGFRneg TICs. For instance, the = expression of=20 other receptor tyrosine kinases, such as PDGFRs, observed in = EGFRneg=20 TIC fractions might contribute to their tumorigenic ability by = synergizing with=20 the EGFR re-upregulation required for experimental gliomagenesis. = Indeed, PDGFR=20 pathway activation has been recently shown to identify a GBM subtype = that does=20 not express the EGFR (46),=20 thus suggesting that the latter GBM subtype might be sustained by=20 EGFRnegPDGFRpos TICs.

EGFRpos and EGFRneg TICs can both = give rise to=20 experimental tumors that phenocopy the tumor of origin, and therefore, = they both=20 can be considered bona fide TICs. However, because EGFRpos = TICs are=20 characterized by enhanced tumorigenic potential and highly invasive = behavior,=20 they might be envisioned as the =E2=80=9Cactual=E2=80=9D GBM CSCs. = Conversely,=20 EGFRneg TICs, which form tumors with low efficiency and need = to=20 re-upregulate the EGFR to be tumorigenic, might be better defined as = =E2=80=9Cpotential=E2=80=9D=20 GBM CSCs, which might be kept in a dormancy-like state by EGFR = downregulation,=20 being reactivated on exposure to appropriate stimuli when exposed to the = in=20 vivo microenvironment. In line with this notion, it has been = recently shown=20 that GBMs, which were EGFRneg in origin, expressed the EGFR = on=20 recurrence (47).=20

Therapeutic implications of EGFR-dependent GBM heterogeneity

The role of EGFR in the malignant progression of GBM and = its effect=20 on patient survival have been highly debated. Despite the controversial=20 prognostic significance of EGFR expression in GBM, pharmacologic = targeting of=20 EGFR by means of TKIs has been proposed as a possible therapeutic = strategy (40).=20

To date, single-agent EGFR inhibition in GBM did not = achieve complete=20 therapeutic efficacy. This failure has been ascribed primarily to the=20 maintenance of a high level of Akt-dependent signaling in PTEN-mutated = tumors=20 (48)=20 and to coactivation in the same tumor cell of multiple receptor tyrosine = kinases, which might compensate for the effective inhibition of = EGFR-dependent=20 pathways (49).=20

By complementing these explanations, our findings suggest = that the=20 simultaneous presence of distinct TICs within the same tumor might also=20 influence the outcome of EGFR-targeted therapies in GBM. Indeed, whereas = effective inhibition of EGFR in our EGFRpos CSC lines can be = achieved=20 independently of PTEN expression, EGFRneg CSCs do not respond = to=20 treatment. By translating these results into a clinical perspective, in = spite of=20 the inhibition of the EGFRpos cell component in = EGFRpos=20 GBM, the nonresponsiveness of the EGFRneg TIC fraction to = EGFR=20 inhibitors might allow these cells to survive treatment and to support = tumor=20 relapse. Therefore, an effective therapeutic regimen should take into=20 consideration the role of residual, nonresponder TICs. To this end, the=20 development of a combination therapy targeted not only against the most=20 aggressive =E2=80=9Cactual=E2=80=9D TIC populations but also against the = less malignant=20 =E2=80=9Cpotential=E2=80=9D TICs will be necessary to improve the = management of GBM.=20

Previous=20 Section= Next=20 Section

Disclosure of Potential Conflicts of Interest

No potential conflicts of interest were = disclosed.

Previous=20 SectionNext=20 Section

Acknowledgments

We thank Alessio Palini and Chiara Villa for technical = assistance=20 with FACS. Antonella Iadanza for technical assistance with MRI, and = Carol=20 Stayton for editing of the manuscript.

Grant Support: Compagnia S. Paolo grants=20 CSP-2002.0735 and CSP-2005.0153, the Oncology Research = Programme-Istituto=20 Superiore di Sanit=C3=A0, and the Fondazione Cariplo (R. Galli).

The costs of publication of this article were defrayed in = part by the=20 payment of page charges. This article must therefore be hereby marked=20 advertisement in accordance with 18 U.S.C. Section 1734 solely = to=20 indicate this fact.

= Previous=20 SectionNext=20 Section

Footnotes

  • Note: Supplementary data for this article = are=20 available at Cancer Research Online (http://cancerres.aacrjournals= .org/).=20

  • Received June 28, 2010.=20
  • Accepted July 18, 2010.
  • =C2=A92010 American = Association for Cancer=20 Research.

Previous=20 Section

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outline-style: none } H1 { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } H2 { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } H3 { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } H4 { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } H5 { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } H6 { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } P { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } BLOCKQUOTE { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = 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outline-style: none } ACRONYM { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } ADDRESS { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } BIG { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } CITE { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } CODE { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } DEL { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } DFN { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } EM { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = 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VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } SMALL { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } STRIKE { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } STRONG { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } SUB { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } SUP { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } TT { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } VAR { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; 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VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } UL { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } LI { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } FIELDSET { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } FORM { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } LABEL { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } LEGEND { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } TABLE { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } CAPTION { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } TBODY { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } TFOOT { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; VERTICAL-ALIGN: baseline; PADDING-TOP: 0px; FONT-FAMILY: = inherit; BORDER-RIGHT-WIDTH: 0px; outline-style: none } THEAD { BORDER-TOP-WIDTH: 0px; PADDING-RIGHT: 0px; PADDING-LEFT: 0px; = BORDER-LEFT-WIDTH: 0px; BORDER-BOTTOM-WIDTH: 0px; PADDING-BOTTOM: 0px; = MARGIN: 0px; 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WIDTH: auto; = POSITION: static; HEIGHT: auto } .main-logo STRONG { BACKGROUND: #fff; FLOAT: none; OVERFLOW: visible; WIDTH: auto; = POSITION: static; HEIGHT: auto } #header .button-list A SPAN { BACKGROUND: #fff; FLOAT: none; OVERFLOW: visible; WIDTH: auto; = POSITION: static; HEIGHT: auto } .secondary-logo { BACKGROUND: #fff; FLOAT: none; OVERFLOW: visible; WIDTH: auto; = POSITION: static; HEIGHT: auto } .secondary-logo STRONG { BACKGROUND: #fff; FLOAT: none; OVERFLOW: visible; WIDTH: auto; = POSITION: static; HEIGHT: auto } .quicksearch LABEL { BACKGROUND: #fff; FLOAT: none; OVERFLOW: visible; WIDTH: auto; = POSITION: static; HEIGHT: auto } .prev-next A SPAN { BACKGROUND: #fff; FLOAT: none; OVERFLOW: visible; WIDTH: auto; = POSITION: static; HEIGHT: auto } #content-block { BORDER-RIGHT-STYLE: none } .pagetype-proxied #content-block { BORDER-RIGHT-STYLE: none } DIV.pagetype-content DIV#content-block H4 { FONT-FAMILY: Times, "Times New Roman", Garamond, serif } ------=_NextPart_000_0000_01CB62F9.AC22C650 Content-Type: text/css; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Content-Location: http://cancerres.aacrjournals.org/content/70/19/ui.css BODY { FONT-SIZE: 100%; MARGIN: 0px auto; WIDTH: 1000px; BACKGROUND-COLOR: = #cccccc } DIV.hw-gen-page { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 10px auto 0px; WIDTH: 960px; COLOR: #403838; PADDING-TOP: 0px; = FONT-FAMILY: "Lucida Sans Unicode", Arial, "Lucida Grande", Tahoma, = Verdana, Helvetica, sans-serif; POSITION: relative; BACKGROUND-COLOR: = white } DIV.hw-gen-page A { COLOR: #202088; TEXT-DECORATION: none } DIV.hw-gen-page A:link { COLOR: #202088; TEXT-DECORATION: none } DIV.hw-gen-page A:visited { COLOR: #581858; TEXT-DECORATION: none } DIV.hw-gen-page A:hover { COLOR: #202088; BORDER-BOTTOM: #202088 1px dotted; TEXT-DECORATION: = none } DIV.hw-gen-page A:active { COLOR: #202088; TEXT-DECORATION: none } #header A { COLOR: #404040; BORDER-TOP-STYLE: none; BORDER-RIGHT-STYLE: none; = BORDER-LEFT-STYLE: none; BORDER-BOTTOM-STYLE: none } #header A:link { COLOR: #404040; BORDER-TOP-STYLE: none; BORDER-RIGHT-STYLE: none; = BORDER-LEFT-STYLE: none; BORDER-BOTTOM-STYLE: none } #footer A { COLOR: #404040; BORDER-TOP-STYLE: none; BORDER-RIGHT-STYLE: none; = BORDER-LEFT-STYLE: none; BORDER-BOTTOM-STYLE: none } #footer A:link { COLOR: #404040; BORDER-TOP-STYLE: none; BORDER-RIGHT-STYLE: none; = BORDER-LEFT-STYLE: none; BORDER-BOTTOM-STYLE: none } #header A:visited { COLOR: #404040; BORDER-TOP-STYLE: none; BORDER-RIGHT-STYLE: none; = BORDER-LEFT-STYLE: none; BORDER-BOTTOM-STYLE: none } #footer A:visited { COLOR: #404040; BORDER-TOP-STYLE: none; BORDER-RIGHT-STYLE: none; = BORDER-LEFT-STYLE: none; BORDER-BOTTOM-STYLE: none } #header UL.button-list A:hover { COLOR: #404040; BORDER-BOTTOM: #404040 1px dotted; TEXT-DECORATION: = none } #header DIV.header-ac-elements A:hover { COLOR: #404040; BORDER-BOTTOM: #404040 1px dotted; TEXT-DECORATION: = none } #header DIV.header-qs A:hover { COLOR: #404040; BORDER-BOTTOM: #404040 1px dotted; TEXT-DECORATION: = none } #footer A:hover { COLOR: #404040; BORDER-BOTTOM: #404040 1px dotted; TEXT-DECORATION: = none } #header A:active { COLOR: #404040; BORDER-TOP-STYLE: none; BORDER-RIGHT-STYLE: none; = BORDER-LEFT-STYLE: none; BORDER-BOTTOM-STYLE: none } #footer A:active { COLOR: #404040; BORDER-TOP-STYLE: none; BORDER-RIGHT-STYLE: none; = BORDER-LEFT-STYLE: none; BORDER-BOTTOM-STYLE: none } #col-2 A { COLOR: #404040 } #col-2 A:link { COLOR: #404040 } #col-3 A { COLOR: #404040 } #col-3 A:link { COLOR: #404040 } #col-2 A:visited { COLOR: #581858 } #col-3 A:visited { COLOR: #581858 } #col-2 A:hover { COLOR: #404040; BORDER-BOTTOM: #404040 1px dotted; TEXT-DECORATION: = none } #col-3 A:hover { COLOR: #404040; BORDER-BOTTOM: #404040 1px dotted; TEXT-DECORATION: = none } #col-2 A:active { COLOR: #404040 } #col-3 A:active { COLOR: #404040 } #col-3 UL.cover-announce A { COLOR: #2f5085 } #col-3 UL.cover-announce A:link { COLOR: #2f5085 } #col-3 UL.cover-announce A:visited { COLOR: #2f5085 } #col-3 UL.cover-announce A:hover { COLOR: #2f5085; BORDER-BOTTOM: #2f5085 1px dotted; TEXT-DECORATION: = none } #col-3 UL.cover-announce A:active { COLOR: #2f5085 } DIV.sidebar A:hover { TEXT-DECORATION: none } DIV#header { BACKGROUND-POSITION: right top; MARGIN-TOP: 0px; BACKGROUND-IMAGE: = url(/shared/img/standard-design/design2/corner.gif); WIDTH: 960px; = BACKGROUND-REPEAT: no-repeat; BORDER-BOTTOM-STYLE: none } #header #main-logo { MARGIN: 15px 0px 0px 24px; BORDER-TOP-STYLE: none; BORDER-RIGHT-STYLE: = none; BORDER-LEFT-STYLE: none; BORDER-BOTTOM-STYLE: none } #header UL.home-page-ads { PADDING-RIGHT: 10px; PADDING-LEFT: 10px; LEFT: 450px; PADDING-BOTTOM: = 5px; MARGIN: 1.5em 0px 35px; WIDTH: 250px; PADDING-TOP: 5px; = LIST-STYLE-TYPE: none; POSITION: absolute; TOP: 0px } #header UL.home-page-ads LI { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; WIDTH: 250px; PADDING-TOP: 0px; HEIGHT: 40px; TEXT-ALIGN: center } #header UL.home-page-ads LI IMG { BORDER-RIGHT: #aaaaaa 1px solid; BORDER-TOP: #aaaaaa 1px solid; = DISPLAY: block; BORDER-LEFT: #aaaaaa 1px solid; WIDTH: 250px; = BORDER-BOTTOM: #aaaaaa 1px solid; HEIGHT: 40px; BACKGROUND-COLOR: = #e0e0e0 } #header .banner-ads { PADDING-RIGHT: 10px; PADDING-LEFT: 10px; LEFT: 450px; PADDING-BOTTOM: = 5px; MARGIN: 1.5em 0px 35px; WIDTH: 250px; PADDING-TOP: 5px; = LIST-STYLE-TYPE: none; POSITION: absolute; TOP: 0px } .banner-ads#header UL { LIST-STYLE-TYPE: none } #header .banner-ads LI { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; PADDING-TOP: 0px; TEXT-ALIGN: center } #header .banner-ads LI IMG { BORDER-RIGHT: #aaaaaa 1px solid; BORDER-TOP: #aaaaaa 1px solid; = DISPLAY: block; BORDER-LEFT: #aaaaaa 1px solid; BORDER-BOTTOM: #aaaaaa = 1px solid; BACKGROUND-COLOR: #e0e0e0 } #header .header-buttons { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: 5px = 0px 8px 24px; PADDING-TOP: 0px; LIST-STYLE-TYPE: none } #header .header-buttons { COLOR: #0083a8 } #header .header-buttons A { COLOR: #0083a8 } #header .header-buttons LI { PADDING-RIGHT: 0.4em; DISPLAY: inline; PADDING-LEFT: 0.4em; = PADDING-BOTTOM: 0px; MARGIN: 0px; BORDER-LEFT: #666666 1px solid; = PADDING-TOP: 0px } #header .header-buttons LI SPAN { FONT-WEIGHT: bold; FONT-SIZE: 87.5% } #header .header-buttons LI.first { PADDING-LEFT: 0px; BORDER-LEFT-STYLE: none } #header .corner { RIGHT: 0px; POSITION: absolute; TOP: 0px } #header .inst-branding { BORDER-LEFT: #c8c8c8 1px dashed } #header #hdr-login { BORDER-LEFT: #c8c8c8 1px dashed } #header .header-qs { BORDER-LEFT: #c8c8c8 1px dashed } #header .inst-branding { PADDING-LEFT: 5px; LEFT: 756px; POSITION: absolute; TOP: 0px; HEIGHT: = 30px } #header #authstring { DISPLAY: block; LEFT: 0%; WIDTH: 756px; LINE-HEIGHT: 100%; PADDING-TOP: = 5px; POSITION: absolute; TEXT-ALIGN: center } #header #authstring { COLOR: white } #header #authstring LI { PADDING-RIGHT: 2px; PADDING-LEFT: 2px; FONT-SIZE: 87.5%; = PADDING-BOTTOM: 2px; BORDER-TOP-STYLE: none; PADDING-TOP: 2px; = BORDER-RIGHT-STYLE: none; BORDER-LEFT-STYLE: none; LIST-STYLE-TYPE: = none; BORDER-BOTTOM-STYLE: none } #header #authstring A { COLOR: white } #header #hdr-login { PADDING-RIGHT: 0px; PADDING-LEFT: 10px; LEFT: 756px; PADDING-BOTTOM: = 5px; PADDING-TOP: 0px; POSITION: absolute; TOP: 30px } #header #hdr-login FORM LABEL { FONT-SIZE: 87.5%; WHITE-SPACE: nowrap } #header #hdr-login INPUT { FONT-SIZE: 87.5%; WHITE-SPACE: nowrap } #header #hdr-login { COLOR: #0083a8 } #header #hdr-login LABEL { PADDING-RIGHT: 0px; DISPLAY: block; PADDING-LEFT: 0px; FONT-SIZE: = 87.5%; PADDING-BOTTOM: 2px; MARGIN: 0px; WIDTH: 80px; PADDING-TOP: 5px } #header #hdr-login INPUT { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; FONT-SIZE: 87.5%; = PADDING-BOTTOM: 0px; WIDTH: 110px; PADDING-TOP: 0px; HEIGHT: 15px } #header .header-qs INPUT { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; FONT-SIZE: 87.5%; = PADDING-BOTTOM: 0px; WIDTH: 110px; PADDING-TOP: 0px; HEIGHT: 15px } #header #hdr-login INPUT#hdr-login-signin { PADDING-RIGHT: 0px; PADDING-LEFT: 5px; PADDING-BOTTOM: 0px; = VERTICAL-ALIGN: bottom; WIDTH: auto; PADDING-TOP: 0px; HEIGHT: auto } #header .bar { PADDING-LEFT: 20px; MARGIN-LEFT: -20px; WIDTH: 980px; HEIGHT: 60px } #footer .bar { PADDING-LEFT: 20px; MARGIN-LEFT: -20px; WIDTH: 980px; HEIGHT: 60px } #header .bar { BACKGROUND-COLOR: #0083a8 } #footer .bar { BACKGROUND-COLOR: #0083a8 } #header .bar-inner { WIDTH: 960px; COLOR: white; HEIGHT: 60px } #header .bar-inner { BACKGROUND-COLOR: #00a0cc } #footer .footer-group { BACKGROUND-COLOR: #00a0cc } #header .header-qs { PADDING-RIGHT: 5px; DISPLAY: block; PADDING-LEFT: 10px; Z-INDEX: 1; = LEFT: 756px; PADDING-BOTTOM: 7px; MARGIN: 0px; WIDTH: 195px; = PADDING-TOP: 10px; POSITION: absolute; TOP: 112px; HEIGHT: 40px } #header .header-qs LABEL { LEFT: -9999px; POSITION: absolute } #header #hdr-login-signin-label { LEFT: -9999px; POSITION: absolute } #header #header-qs-search-label { LEFT: -9999px; POSITION: absolute } #header .header-qs A { FONT-SIZE: 87.5%; COLOR: white; LINE-HEIGHT: 1.5; WHITE-SPACE: nowrap } #header DIV.adv-search-link A:hover { COLOR: white; BORDER-BOTTOM: white 1px dotted } #header DIV.adv-search-link A:visited { COLOR: white } #header DIV.adv-search-link A:active { COLOR: white } #header DIV.adv-search-link A:link { COLOR: white } #header .header-qs INPUT#header-qs-search-go { MARGIN-LEFT: 0.5em; VERTICAL-ALIGN: bottom; WIDTH: 27px; HEIGHT: auto } #content-block { PADDING-RIGHT: 20px; PADDING-LEFT: 5px; FLOAT: left; PADDING-BOTTOM: = 0px; WIDTH: 518px; PADDING-TOP: 0px } #col-2 { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; PADDING-TOP: 0px } #col-3 { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; PADDING-TOP: 0px } #col-2 { BORDER-RIGHT: #aaaaaa 1px dashed; PADDING-RIGHT: 13px; FLOAT: left; = MARGIN: 0px; OVERFLOW: hidden; WIDTH: 200px } #content-block { COLOR: #403838; BACKGROUND-COLOR: white } .pagetype-proxied #content-block { BORDER-RIGHT: #aaaaaa 1px dashed; PADDING-RIGHT: 0px; PADDING-LEFT: = 0px; PADDING-BOTTOM: 0px; WIDTH: 756px; PADDING-TOP: 0px } .proxied-column-display#proxied-contents #col-main { PADDING-RIGHT: 10px; FLOAT: left; OVERFLOW: hidden; WIDTH: 518px } .proxied-column-display#proxied-contents #col-2 { BORDER-RIGHT: 0px solid; FLOAT: left } #proxied-contents { PADDING-RIGHT: 10px; PADDING-LEFT: 15px } .proxied-column-display#proxied-contents { PADDING-RIGHT: 0px } .proxied-column-display#proxied-contents .search-nav A { FONT-WEIGHT: normal; COLOR: black } .pagetype-proxied #header .banner-ads UL { MARGIN: 1em 5% } .pagetype-proxied #col-3 .content-box H3 { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; FONT-SIZE: 1.2em; = PADDING-BOTTOM: 0.3em; MARGIN: 0px; PADDING-TOP: 0.3em } #proxied-contents TH { FONT-WEIGHT: bold } #proxied-contents TABLE { LINE-HEIGHT: 110% } #proxied-contents TD { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; PADDING-TOP: 0px } TR { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; PADDING-TOP: 0px } TH { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; PADDING-TOP: 0px } P { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; PADDING-TOP: 0px } OL { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; PADDING-TOP: 0px } UL { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; PADDING-TOP: 0px } #proxied-contents H1 { LINE-HEIGHT: 120%; TEXT-ALIGN: left } H3 { LINE-HEIGHT: 120%; TEXT-ALIGN: left } DL { LINE-HEIGHT: 120%; TEXT-ALIGN: left } DT { LINE-HEIGHT: 120%; TEXT-ALIGN: left } DD { LINE-HEIGHT: 120%; TEXT-ALIGN: left } #proxied-contents .proxied-collections H1 { MARGIN-TOP: 0.5em } #col-3 H1 { LINE-HEIGHT: 100%; TEXT-ALIGN: left } H3 { LINE-HEIGHT: 100%; TEXT-ALIGN: left } DL { LINE-HEIGHT: 100%; TEXT-ALIGN: left } DT { LINE-HEIGHT: 100%; TEXT-ALIGN: left } DD { LINE-HEIGHT: 100%; TEXT-ALIGN: left } #proxied-contents #col-3 P { MARGIN: 1em 5% } PRE { MARGIN: 1em 5% } LABEL { MARGIN: 1em 5% } UL { MARGIN: 1em 5% } OL { MARGIN: 1em 5% } DL { MARGIN: 1em 5% } FIELDSET { MARGIN: 1em 5% } ADDRESS { MARGIN: 1em 5% } #col-2 #cb-art-nav LI { MARGIN-TOP: 0.4em } #col-2 DIV.sidebar-issue-nav { MARGIN: 5px 0px 0px 5px } #col-2 DIV.article-nav { MARGIN: 5px 0px 0px 5px } #col-2 DIV.search-nav { MARGIN: 5px 0px 0px 5px } #col-3 { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; FLOAT: left; PADDING-BOTTOM: = 0px; MARGIN: 0px; OVERFLOW: visible; WIDTH: 203px; PADDING-TOP: 0px; = TEXT-ALIGN: left } #col-3 { BACKGROUND-COLOR: white } #col-3 * { POSITION: static } #col-3 DIV { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; PADDING-BOTTOM: 0px; MARGIN: = 0px; PADDING-TOP: 0px } DIV.content-box A:link { COLOR: #404040; TEXT-DECORATION: none } DIV.content-box A:visited { COLOR: #404040; TEXT-DECORATION: none } DIV.content-box A:hover { COLOR: #404040; TEXT-DECORATION: none } DIV.content-box A:active { COLOR: #404040; TEXT-DECORATION: none } #col-3 UL.pub-links LI A:link { BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none } #col-3 UL.pub-links LI A:visited { BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none } #col-3 UL.pub-links LI A:hover { BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none } #col-3 UL.pub-links LI A:active { BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none } #col-3 UL.ex-pub-links LI A:link { BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none } #col-3 UL.ex-pub-links LI A:visited { BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none } #col-3 UL.ex-pub-links LI A:hover { BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none } #col-3 UL.ex-pub-links LI A:active { BORDER-BOTTOM-WIDTH: 0px; TEXT-DECORATION: none } #col-3 UL.pub-links { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; LEFT: -7px; PADDING-BOTTOM: 0px; = MARGIN: 10px 0px; OVERFLOW: hidden; PADDING-TOP: 0px; POSITION: relative } #col-3 UL.pub-links { LIST-STYLE-TYPE: none } #col-3 UL.pub-links { COLOR: #0083a8 } #col-3 UL.pub-links LI A { COLOR: #0083a8 } #col-3 UL.pub-links LI { BORDER-RIGHT: #aaaaaa 1px dashed; PADDING-RIGHT: 5px; BORDER-TOP: = #aaaaaa 1px dashed; PADDING-LEFT: 4px; FONT-WEIGHT: bold; FONT-SIZE: = 0.85em; PADDING-BOTTOM: 10px; MARGIN: 0px; BORDER-LEFT: #aaaaaa 1px = dashed; PADDING-TOP: 10px; BORDER-BOTTOM: #aaaaaa 1px; POSITION: static; = BACKGROUND-COLOR: white } #col-3 UL.pub-links LI A { BACKGROUND-POSITION: left top; DISPLAY: block; PADDING-LEFT: 1em; = BACKGROUND-IMAGE: = url(/shared/img/standard-design/design2/jnl-link-bullet.gif); = LINE-HEIGHT: 1.2; BACKGROUND-REPEAT: no-repeat } #col-3 UL.pub-links LI.last { BORDER-BOTTOM: #aaaaaa 1px dashed } #col-3 .sidebar-icon-group { PADDING-RIGHT: 20px; PADDING-LEFT: 20px; PADDING-BOTTOM: 20px; = PADDING-TOP: 20px } #col-3 .sidebar-icon-group A:hover { BORDER-BOTTOM: 0px solid } #col-3 UL.ex-pub-links { PADDING-RIGHT: 0px; PADDING-LEFT: 0px; LEFT: -7px; PADDING-BOTTOM: 0px; = MARGIN: 10px 0px; OVERFLOW: hidden; PADDING-TOP: 0px; POSITION: relative } #col-3 UL.ex-pub-links { LIST-STYLE-TYPE: none } #col-3 UL.ex-pub-links { COLOR: #0083a8 } #col-3 UL.ex-pub-links LI A { COLOR: #0083a8 } #col-3 UL.ex-pub-links LI { BORDER-RIGHT: #aaaaaa 1px dashed; PADDING-RIGHT: 5px; BORDER-TOP: = #aaaaaa 1px dashed; PADDING-LEFT: 4px; FONT-WEIGHT: bold; FONT-SIZE: = 75%; PADDING-BOTTOM: 10px; MARGIN: 0px; BORDER-LEFT: #aaaaaa 1px dashed; = PADDING-TOP: 10px; BORDER-BOTTOM: #aaaaaa 1px; POSITION: static; = BACKGROUND-COLOR: white } #col-3 UL.ex-pub-links LI A { BACKGROUND-POSITION: left top; DISPLAY: block; PADDING-LEFT: 1em; = BACKGROUND-IMAGE: = url(/shared/img/standard-design/design2/jnl-link-bullet.gif); = LINE-HEIGHT: 1.2; BACKGROUND-REPEAT: no-repeat } #col-3 UL.ex-pub-links LI.last { BORDER-BOTTOM: #aaaaaa 1px dashed } #col-3 UL.tower-ads { PADDING-RIGHT: 0em; PADDING-LEFT: 0em; 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        • ';=0A= }=0A= }=0A= );=0A= }=0A= hdrList +=3D '
      ';=0A= if (hasItems) {=0A= var header =3D $this.find("div.most-header h3");=0A= if (header.length) {=0A= header.after(hdrList);=0A= }=0A= }=0A= mostHdrList =3D $this.find("div.most-header ul");=0A= }=0A= // 3) if none of the items are selected, make the first one be=0A= if (((mostHdrList !=3D null) && mostHdrList.length) && = (!hasSelected)) {=0A= modClass(mostHdrList.find("li:first"), "most-cur-sel", "");=0A= modClass($this.children("ul").children("li:first"), "most-cur-sel", = "");=0A= }=0A= // 4) bind a handler to each heading li a to toggle = class=3D"most-cur-sel" for =0A= // that item and its corresponding item in the list below=0A= if (((mostHdrList !=3D null) && mostHdrList.length)) {=0A= mostHdrList.find("li a").click(=0A= function(e) {=0A= var curA =3D $(this);=0A= toggleMostSelection(e, $this, curA.text());=0A= }=0A= );=0A= }=0A= }=0A= );=0A= if (hadMostBoxes) {=0A= fixColHeights(-1);=0A= }=0A= }=0A= =0A= function toggleMostSelection(e, mostItemDiv, groupName) {=0A= if (mostItemDiv.length) {=0A= var hdrItems =3D mostItemDiv.find("div.most-header ul li");=0A= if (hdrItems.length) {=0A= hdrItems.each(=0A= function() {=0A= var hdrName =3D $(this).text();=0A= var isChoice =3D (hdrName =3D=3D groupName);=0A= if (isChoice) {=0A= modClass($(this), 'most-cur-sel', '');=0A= }=0A= else {=0A= modClass($(this), '', 'most-cur-sel');=0A= }=0A= }=0A= );=0A= }=0A= var listItems =3D mostItemDiv.children("ul").find("li");=0A= if (listItems.length) {=0A= listItems.each(=0A= function() {=0A= var itemName =3D $(this).children("h4").text();=0A= var isChoice =3D (itemName =3D=3D groupName);=0A= if (isChoice) {=0A= modClass($(this), 'most-cur-sel', '');=0A= }=0A= else {=0A= modClass($(this), '', 'most-cur-sel');=0A= }=0A= }=0A= );=0A= }=0A= }=0A= fixColHeights(-1);=0A= e.preventDefault();=0A= }=0A= =0A= function toggleCollapse(collapsibleEl, hideTagExpr, modTextTagExpr, = expandStr, contractStr) {=0A= if (typeof(expandStr) =3D=3D "undefined") {=0A= expandStr =3D getSiteOption('expandString','+');=0A= }=0A= if (typeof(contractStr) =3D=3D "undefined") {=0A= contractStr =3D getSiteOption('contractString','-');=0A= }=0A= =0A= var $this =3D collapsibleEl;=0A= if ($this.hasClass('collapsed')) {=0A= modClass($this, 'collapsible', 'collapsed');=0A= }=0A= else if ($this.hasClass('collapsible')) {=0A= modClass($this, 'collapsed', 'collapsible');=0A= }=0A= var $toToggle =3D $this.find(hideTagExpr);=0A= if ($toToggle.length) {=0A= $toToggle.toggle();=0A= }=0A= var $rewrite =3D $this.find(modTextTagExpr);=0A= if ($rewrite.length) {=0A= $rewrite.each(=0A= function(i, e) {=0A= var txt =3D $(e).text();=0A= if (txt =3D=3D expandStr) {=0A= $(e).empty().append(contractStr);=0A= }=0A= else if (txt =3D=3D contractStr) {=0A= $(e).empty().append(expandStr);=0A= }=0A= }=0A= );=0A= }=0A= }=0A= =0A= function setupDockBlock(col, dockId, dockClass, contentRules) {=0A= if ((col =3D=3D 2) || (col =3D=3D 3)) {=0A= var navOrigHeight =3D ((col =3D=3D 2) ? gColOrigHeights.col2 : = gColOrigHeights.col3);=0A= var gColNum =3D col;=0A= var gNav =3D ((col =3D=3D 2) ? gCol2 : gCol3);=0A= var gNavHeight =3D ((gNav.length) ? navOrigHeight : 0);=0A= var gNavTopOffset =3D (gNavHeight > 0 ? gNav.offset().top : 0);=0A= var gScrollPos =3D 0;=0A= var gNavDocked =3D false;=0A= /* this won't work in IE 6, so skip it */=0A= /* article navigation docking block */=0A= $(window).scroll(function () {=0A= if (gNavHeight > 0 && !isIE6) {=0A= var offsets =3D getPageOffset();=0A= var origHeight =3D ((gColNum =3D=3D 2) ? gColOrigHeights.col2 : = gColOrigHeights.col3);=0A= var navBottom =3D (gNavTopOffset + origHeight) - offsets.y;=0A= if (gScrollPos !=3D offsets.y) {=0A= gScrollPos =3D offsets.y;=0A= if ((navBottom <=3D 0) && !gNavDocked) {=0A= /* dock the nav */=0A= gNavDocked =3D true;=0A= addDockBlock(col, dockId, dockClass, contentRules);=0A= addDockBlockCallback(dockId);=0A= }=0A= else if (gNavDocked && (navBottom > 0)) {=0A= /* undock the nav */=0A= gNavDocked =3D false;=0A= removeDockBlock(col, dockId);=0A= removeDockBlockCallback(dockId);=0A= }=0A= }=0A= }=0A= });=0A= }=0A= }=0A= function addDockBlock(col, dockId, dockClass, contentRules) {=0A= // get direct children li elements only=0A= var colId =3D ("col-" + col);=0A= var newDiv =3D ' 1) {=0A= for (var i =3D 0; i < (contentRules.length - 1); i =3D i + 2) {=0A= var addTo =3D processDockingRule(contentRules[i]);=0A= var add =3D processDockingRule(contentRules[i + 1]);=0A= var addTarget, addVal;=0A= if (addTo.ruleType =3D=3D 'empty') {=0A= addTarget =3D newDivJQuery;=0A= }=0A= else if (addTo.ruleType =3D=3D 'jQuery') {=0A= addTarget =3D $(addTo.rule);=0A= }=0A= if (add.ruleType =3D 'jQuery') {=0A= var addJQEl =3D $(add.rule);=0A= if (addJQEl.length) {=0A= addVal =3D addJQEl.clone();=0A= }=0A= else {=0A= addVal =3D '';=0A= }=0A= }=0A= else if (add.ruleType =3D 'html') {=0A= addVal =3D add.rule;=0A= }=0A= if (addTarget.length && !(addVal =3D=3D '')) {=0A= addTarget.append(addVal);=0A= }=0A= else {=0A= }=0A= }=0A= }=0A= var newDockJQrule =3D "#" + colId + " #" + dockId;=0A= var newDock =3D $(newDockJQrule);=0A= if (newDock.length) {=0A= prepCollapsibles(newDockJQrule + " div.collapsible, " + newDockJQrule = + " div.collapsed", false);=0A= newDock.fadeIn(250);=0A= }=0A= }=0A= =0A= function processDockingRule(rule) {=0A= var retObj =3D new Object();=0A= retObj.empty =3D (((rule !=3D undefined) && (!(rule =3D=3D ''))) ? = false : true);=0A= if (rule =3D=3D '') {=0A= retObj.ruleType =3D 'empty';=0A= }=0A= else if (/^\$\(.+\)$/.test(rule)) {=0A= retObj.ruleType =3D 'jQuery';=0A= retObj.rule =3D rule.replace(/^\$\((.+)\)$/,"$1");=0A= }=0A= else {=0A= retObj.ruleType =3D 'html';=0A= retObj.rule =3D rule;=0A= }=0A= return retObj;=0A= }=0A= =0A= function removeDockBlock(col, dockId) {=0A= var dockedNav =3D $("div#" + dockId);=0A= if(dockedNav.length) {=0A= dockedNav.fadeOut(250, function() { dockedNav.remove(); });=0A= }=0A= }=0A= // custom implementations can override this function to do something = when a dockBlock is added=0A= // they should return false if they want to stop any additional default = handling=0A= function customAddDockBlockCallback(dockId) {=0A= return true;=0A= }=0A= // this function should not be overridden=0A= function addDockBlockCallback(dockId) {=0A= if (customAddDockBlockCallback(dockId) !=3D false) {=0A= // no default handling, currently=0A= }=0A= }=0A= // custom implementations can override this function to do something = when a dockBlock is removed=0A= // they should return false if they want to stop any additional default = handling=0A= function customRemoveDockBlockCallback(dockId) {=0A= return true;=0A= }=0A= // this function should not be overridden=0A= function removeDockBlockCallback(dockId) {=0A= if (customRemoveDockBlockCallback(dockId) !=3D false) {=0A= // no default handling, currently=0A= }=0A= }=0A= =0A= function updateFormInput(labelMatchString, inputMatchString, = defaultColorString, textColorString) {=0A= if ((defaultColorString =3D=3D null) || (defaultColorString =3D=3D '')) = {=0A= defaultColorString =3D "#A0A0A0";=0A= }=0A= if ((textColorString =3D=3D null) || (textColorString =3D=3D '')) {=0A= textColorString =3D "black";=0A= }=0A= var label =3D $(labelMatchString);=0A= var input =3D $(inputMatchString);=0A= if (input.length) {=0A= if ((label.length) && (input.val() =3D=3D '')) {=0A= input.val(label.text()).css("color",defaultColorString);=0A= }=0A= input.focus(=0A= function(e) {=0A= if ((label.length) && ($(this).val() =3D=3D label.text())) {=0A= $(this).val('').css("color",textColorString);=0A= }=0A= }=0A= );=0A= input.blur(=0A= function(e) {=0A= if ((label.length) && ($(this).val() =3D=3D '')) {=0A= $(this).val(label.text()).css("color",defaultColorString);=0A= }=0A= }=0A= );=0A= }=0A= var parentForm =3D label.parents("form").eq(0);=0A= if (parentForm.length) {=0A= parentForm.submit(=0A= function() {=0A= if ((label.length) && (input.length) && (input.val() =3D=3D = label.text())) {=0A= input.val('');=0A= }=0A= return true;=0A= }=0A= );=0A= }=0A= }=0A= =0A= function checkUnloadedImgs(lookupRule) {=0A= if (lookupRule =3D=3D null) {=0A= lookupRule =3D "img";=0A= }=0A= var imageNum =3D -1;=0A= var images =3D $(lookupRule);=0A= if (images.length) {=0A= imageNum =3D 0;=0A= images.each(=0A= function() {=0A= //if ($(this).height() =3D=3D 0) {=0A= if (this.offsetHeight =3D=3D 0) {=0A= imageNum++;=0A= }=0A= }=0A= );=0A= }=0A= return imageNum;=0A= }=0A= =0A= function fixHeightForImages(iter, numImagesToLoad, lookupRule) {=0A= if (lookupRule =3D=3D null) {=0A= lookupRule =3D "img";=0A= }=0A= if (numImagesToLoad > 0) {=0A= var lastNumImagesToLoad =3D numImagesToLoad;=0A= var newNumImagesToLoad =3D checkUnloadedImgs(lookupRule);=0A= if (newNumImagesToLoad < lastNumImagesToLoad) {=0A= fixColHeights(1);=0A= if ((newNumImagesToLoad > 0) && (iter < 10)) {=0A= setTimeout("fixHeightForImages(" + (iter+1) + "," + = newNumImagesToLoad + ",'" + lookupRule + "')", 1000);=0A= }=0A= }=0A= }=0A= }=0A= function getColInfo(colJQEl) {=0A= var colInfo =3D { valid: false, height: 0, bottom: 0, lastElBottom: 0, = extra: 0, fix: false };=0A= if (colJQEl.length) {=0A= var c =3D colJQEl.children();=0A= if (c.length) {=0A= colInfo.height =3D getObjHeight(colJQEl);=0A= colInfo.bottom =3D colJQEl.offset().top + colInfo.height;=0A= var last =3D c.eq(c.length - 1);=0A= colInfo.lastElBottom =3D last.offset().top + getObjHeight(last);=0A= colInfo.extra =3D colInfo.bottom - colInfo.lastElBottom;=0A= colInfo.valid =3D true;=0A= }=0A= }=0A= return colInfo;=0A= }=0A= function checkColHeights() {=0A= var colToFix =3D 0;=0A= if ((gIsMultiCol || gColTempResize) && ((gColOrigHeights.main + = gColOrigHeights.col2 + gColOrigHeights.col3) > 0)) {=0A= var colMainInfo =3D getColInfo(gColMain);=0A= var col2Info =3D getColInfo(gCol2);=0A= var col3Info =3D getColInfo(gCol3);=0A= if (colMainInfo.valid && (gColLows.main =3D=3D 0)) { gColLows.main =3D = colMainInfo.lastElBottom; }=0A= if (col2Info.valid && (gColLows.col2 =3D=3D 0)) { gColLows.col2 =3D = col2Info.lastElBottom; }=0A= if (col3Info.valid && (gColLows.col3 =3D=3D 0)) { gColLows.col3 =3D = col3Info.lastElBottom; }=0A= // check if low point in any column has changed=0A= if ((colMainInfo.valid) && (colMainInfo.lastElBottom !=3D = gColLows.main)) {=0A= colMainInfo.fix =3D true;=0A= }=0A= if ((col2Info.valid) && (col2Info.lastElBottom !=3D gColLows.col2)) {=0A= col2Info.fix =3D true;=0A= }=0A= if ((col3Info.valid) && (col3Info.lastElBottom !=3D gColLows.col3)) {=0A= col3Info.fix =3D true;=0A= }=0A= // check for columns not having same bottom points=0A= if ((colMainInfo.valid) &&=0A= (((col2Info.valid) && (colMainInfo.height < col2Info.height)) ||=0A= ((col3Info.valid) && (colMainInfo.height < col3Info.height)))) {=0A= colMainInfo.fix =3D true;=0A= }=0A= if ((col2Info.valid) &&=0A= (((colMainInfo.valid) && (col2Info.height < colMainInfo.height)) ||=0A= ((col3Info.valid) && (col2Info.height < col3Info.height)))) {=0A= col2Info.fix =3D true;=0A= }=0A= if ((col3Info.valid) &&=0A= (((colMainInfo.valid) && (col3Info.height < colMainInfo.height)) ||=0A= ((col2Info.valid) && (col3Info.height < col2Info.height)))) {=0A= col3Info.fix =3D true;=0A= }=0A= =0A= if (colToFix > -1) {=0A= if (colMainInfo.valid && colMainInfo.fix) {=0A= if ((col2Info.valid && col2Info.fix) || (col3Info.valid && = col3Info.fix)) {=0A= colToFix =3D -1;=0A= }=0A= else { colToFix =3D 1; }=0A= }=0A= else if (col2Info.valid && col2Info.fix) {=0A= if (col3Info.valid && col3Info.fix) {=0A= colToFix =3D -1;=0A= }=0A= else { colToFix =3D 2; }=0A= }=0A= else if (col3Info.valid && col3Info.fix) {=0A= colToFix =3D 3;=0A= }=0A= }=0A= =0A= if (colToFix !=3D 0) {=0A= fixColHeights(colToFix);=0A= }=0A= if (colMainInfo.valid) { gColLows.main =3D colMainInfo.lastElBottom; }=0A= if (col2Info.valid) { gColLows.col2 =3D col2Info.lastElBottom; }=0A= if (col3Info.valid) { gColLows.col3 =3D col3Info.lastElBottom; }=0A= }=0A= setTimeout("checkColHeights()", 5000);=0A= }=0A= function fixColHeights(colChanged) {=0A= if ((gIsMultiCol || gColTempResize) && getSiteOption('setColSizes', = true)) {=0A= var setAll =3D false;=0A= if (colChanged =3D=3D 1) {=0A= if (gColMain.length) {=0A= setObjHeight(gColMain,'auto');=0A= gColOrigHeights.main =3D getObjHeight(gColMain);=0A= }=0A= }=0A= else if (colChanged =3D=3D 2) {=0A= if (gCol2.length) {=0A= setObjHeight(gCol2,'auto');=0A= gColOrigHeights.col2 =3D getObjHeight(gCol2);=0A= }=0A= }=0A= else if (colChanged =3D=3D 3) {=0A= if (gCol3.length) {=0A= setObjHeight(gCol3,'auto');=0A= gColOrigHeights.col3 =3D getObjHeight(gCol3);=0A= }=0A= }=0A= else {=0A= if (colChanged =3D=3D -1) {=0A= // force resize all 3=0A= if (gColMain.length) { setObjHeight(gColMain,'auto'); }=0A= if (gCol2.length) { setObjHeight(gCol2,'auto'); }=0A= if (gCol3.length) { setObjHeight(gCol3,'auto'); }=0A= }=0A= gColOrigHeights.main =3D ((gColMain.length) ? = (getObjHeight(gColMain)) : 0);=0A= gColOrigHeights.col2 =3D ((gCol2.length) ? (getObjHeight(gCol2)) : 0);=0A= gColOrigHeights.col3 =3D ((gCol3.length) ? (getObjHeight(gCol3)) : 0);=0A= setAll =3D true;=0A= }=0A= if ((gColOrigHeights.main + gColOrigHeights.col2 + = gColOrigHeights.col3) > 0) {=0A= var colInfo;=0A= var maxH =3D Math.max(Math.max(gColOrigHeights.main, = gColOrigHeights.col2), gColOrigHeights.col3);=0A= if ((gColMain.length) && ((gColOrigHeights.main < maxH) || = (getObjHeight(gColMain) > maxH) || setAll)) {=0A= setObjHeight(gColMain, maxH);=0A= colInfo =3D getColInfo(gColMain);=0A= if (colInfo.valid) { gColLows.main =3D colInfo.lastElBottom; }=0A= }=0A= if ((gCol2.length) && ((gColOrigHeights.col2 < maxH) || = (getObjHeight(gCol2) > maxH) || setAll)) {=0A= setObjHeight(gCol2, maxH);=0A= colInfo =3D getColInfo(gCol2);=0A= if (colInfo.valid) { gColLows.col2 =3D colInfo.lastElBottom; }=0A= }=0A= if ((gCol3.length) && ((gColOrigHeights.col3 < maxH) || = (getObjHeight(gCol3) > maxH) || setAll)) {=0A= setObjHeight(gCol3, maxH);=0A= colInfo =3D getColInfo(gCol3);=0A= if (colInfo.valid) { gColLows.col3 =3D colInfo.lastElBottom; }=0A= }=0A= }=0A= }=0A= }=0A= =0A= function getObjHeight(obj) {=0A= if (obj instanceof jQuery) {=0A= return obj.get(0).offsetHeight;=0A= }=0A= else if (obj instanceof Array) {=0A= return obj[0].offsetHeight;=0A= }=0A= else {=0A= return obj.offsetHeight;=0A= }=0A= }=0A= function getObjWidth(obj) {=0A= if (obj instanceof jQuery) {=0A= return obj.get(0).offsetWidth;=0A= }=0A= else if (obj instanceof Array) {=0A= return obj[0].offsetWidth;=0A= }=0A= else {=0A= return obj.offsetWidth;=0A= }=0A= }=0A= function setObjHeight(obj, h) {=0A= if (typeof(h) =3D=3D 'number') {=0A= h =3D "" + h + "px";=0A= }=0A= if (obj instanceof jQuery) {=0A= obj.get(0).style.height =3D h;=0A= }=0A= else if (obj instanceof Array) {=0A= obj[0].style.height =3D h;=0A= }=0A= else {=0A= obj.style.height =3D h;=0A= }=0A= }=0A= function setObjWidth(obj, w) {=0A= if (typeof(w) =3D=3D 'number') {=0A= w =3D "" + w + "px";=0A= }=0A= if (obj instanceof jQuery) {=0A= obj.get(0).style.width =3D w;=0A= }=0A= else if (obj instanceof Array) {=0A= obj[0].style.width =3D w;=0A= }=0A= else {=0A= obj.style.width =3D w;=0A= }=0A= }=0A= =0A= /* num pixels top of page has been scrolled offscreen */=0A= function getPageOffset() {=0A= var offset =3D new Object();=0A= if( typeof( window.pageXOffset ) =3D=3D 'number' ) {=0A= offset.x =3D window.pageXOffset;=0A= offset.y =3D window.pageYOffset;=0A= } else if( document.body && ( document.body.scrollTop ) ) {=0A= offset.x =3D document.body.scrollLeft;=0A= offset.y =3D document.body.scrollTop;=0A= } else if( document.documentElement && ( = document.documentElement.scrollTop ) ) {=0A= offset.x =3D document.documentElement.scrollLeft;=0A= offset.y =3D document.documentElement.scrollTop;=0A= }=0A= if (typeof(offset.x)=3D=3D"undefined") { offset.x =3D 0; }=0A= if (typeof(offset.y)=3D=3D"undefined") { offset.y =3D 0; }=0A= return offset;=0A= }=0A= function getViewportDim() {=0A= var dim =3D new Object();=0A= // non-IE=0A= if (typeof window.innerWidth !=3D 'undefined') {=0A= dim.x =3D window.innerWidth;=0A= dim.y =3D window.innerHeight;=0A= }=0A= // IE6 in standards compliant mode=0A= else if (typeof document.documentElement !=3D 'undefined'=0A= && typeof document.documentElement.clientWidth !=3D=0A= 'undefined' && document.documentElement.clientWidth !=3D 0) {=0A= dim.x =3D document.documentElement.clientWidth;=0A= dim.y =3D document.documentElement.clientHeight;=0A= }=0A= // older versions of IE=0A= else {=0A= dim.x =3D document.getElementsByTagName('body')[0].clientWidth;=0A= dim.y =3D document.getElementsByTagName('body')[0].clientHeight;=0A= }=0A= return dim;=0A= }=0A= =0A= function newWindowTargets() {=0A= var newWins =3D $("a.in-nw");=0A= if (newWins.length) {=0A= newWins.each(=0A= function() {=0A= var $this =3D $(this);=0A= $this.attr("target", "_blank");=0A= modClass($this, 'in-nw-vis', 'in-nw');=0A= }=0A= );=0A= }=0A= }=0A= =0A= function getSiteOption(optionName, defaultVal) {=0A= if (typeof(defaultVal) =3D=3D "undefined") {=0A= defaultVal =3D gSiteOptionUnknown;=0A= }=0A= if (gSiteOptions =3D=3D undefined) {=0A= return defaultVal;=0A= }=0A= else if (gSiteOptions[optionName] =3D=3D undefined) {=0A= return defaultVal;=0A= }=0A= else {=0A= return gSiteOptions[optionName];=0A= }=0A= }=0A= =0A= function addPartHeaders(req) {=0A= req.setRequestHeader('Accept', 'application/xhtml+xml');=0A= req.setRequestHeader('Range', 'part=3Dvariant-contents');=0A= }=0A= function addCommonHeaders(req) {=0A= if (typeof(callbackToken) !=3D 'undefined') {=0A= req.setRequestHeader('X-Token', callbackToken);=0A= }=0A= }=0A= function allowsCookies() {=0A= if (hasTestCookie()) {=0A= return true;=0A= }=0A= else {=0A= setTestCookie();=0A= return hasTestCookie();=0A= }=0A= }=0A= function setTestCookie() {=0A= setCookie('cks','allowed',7,'/',null,null);=0A= }=0A= function hasTestCookie() {=0A= var ckVal =3D getCookie('cks');=0A= if ((ckVal !=3D null) && (ckVal =3D=3D 'allowed')) {=0A= return true;=0A= }=0A= else {=0A= return false;=0A= }=0A= }=0A= =0A= var UIPrefsCk=3D{=0A= name: "UIPrefs",=0A= expDays: 3652,=0A= path: '/'=0A= };=0A= =0A= function prefDefined(name) {=0A= var val =3D getPrefValue(name);=0A= if (val =3D=3D null) {=0A= return false;=0A= }=0A= else {=0A= return true;=0A= }=0A= }=0A= function getPrefValue(name) {=0A= var prefArray =3D getPrefArray();=0A= if (prefArray) {=0A= name =3D convertPrefString(name);=0A= var match =3D name + ":";=0A= for (var i =3D 0; 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      This Article

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'&' : '?') = + 'baseURI=3D' + ((baseAjaxUrl.indexOf('?') > 0) ? = baseAjaxUrl.substring(0, baseAjaxUrl.indexOf('?')): baseAjaxUrl);=0A= //var ajaxUrl =3D baseAjaxUrl + ((href.indexOf('?') >=3D 0) ? '&' : = '?') + 'baseURI=3D' + baseAjaxUrl;=0A= $.ajax({=0A= url: ajaxUrl,=0A= dataType: "html",=0A= type: "GET",=0A= error: ajaxErr,=0A= beforeSend: addFigHeaders,=0A= success: function(xhtml) {=0A= addFig(xhtml, figWrapperEl);=0A= },=0A= complete: ajaxComplete=0A= });=0A= e.preventDefault();=0A= }=0A= function addFigHeaders(req) {=0A= addCommonHeaders(req);=0A= addPartHeaders(req);=0A= }=0A= =0A= function addFig(xhtmlData, figWrapperEl) {=0A= if (xhtmlData && !(xhtmlData.indexOf('=3D 0)) {=0A= figWrapperEl.replaceWith(xhtmlData);=0A= newWindowTargets();=0A= var lookupRule =3D "div.article div.fig img";=0A= var numImagesToLoad =3D checkUnloadedImgs(lookupRule);=0A= setTimeout("fixHeightForImages(1" + "," + numImagesToLoad + ",'" + = lookupRule + "')", 1000);=0A= fixColHeights(1);=0A= }=0A= }=0A= =0A= function refLinksNewWindowTarget() {=0A= $("div.ref-list div.cit-extra a").each(=0A= function(i) {=0A= var origTitle =3D $(this).attr("title");=0A= var newTitle =3D '';=0A= if ((origTitle =3D=3D undefined) || (!origTitle)) {=0A= origTitle =3D '';=0A= }=0A= else {=0A= newTitle =3D origTitle + ' ';=0A= }=0A= newTitle +=3D '[opens in a new window]';=0A= $(this).attr("target", "_blank").attr("title", newTitle);=0A= }=0A= );=0A= }=0A= =0A= function addRefPops() {=0A= var numMissed =3D 0;=0A= var maxToSkip =3D getSiteOption('refPopsMaxToSkip', 10);=0A= var i =3D 1;=0A= var idroot =3D "#xref-ref-";=0A= var el =3D $(idroot + i + '-1');=0A= /* not all refs appear in text; compensate */=0A= while (numMissed < maxToSkip) {=0A= if (el.length) {=0A= numMissed =3D 0;=0A= el.hover(dispRef, hideRef);=0A= var j =3D 2;=0A= var el2 =3D $(idroot + i + "-" + j);=0A= while (el2.length) {=0A= el2.hover(dispRef, hideRef);=0A= j++;=0A= el2 =3D $(idroot + i + "-" + j);=0A= }=0A= }=0A= else {=0A= numMissed++;=0A= }=0A= i++;=0A= el =3D $(idroot + i + "-1");=0A= }=0A= }=0A= =0A= function dispRef(e) {=0A= var link =3D $(this).attr("href");=0A= if($("div#hovering-ref").length) {=0A= $("div#hovering-ref").remove();=0A= //alert("hovering-ref div removed on new hover!");=0A= }=0A= var linkEl =3D $(link);=0A= if (linkEl.length) {=0A= var citHtml =3D linkEl.next("div").children("div.cit-metadata");=0A= if (!(citHtml.length)) {=0A= citHtml =3D linkEl.parent().next("div").children("div.cit-metadata");=0A= }=0A= if (citHtml.length) {=0A= var newDiv =3D '
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        ';=0A= $("body").append(newDiv);=0A= var elH =3D getObjHeight($("div#hovering-ref"));=0A= $("div#hovering-ref").css("left", e.pageX+10).css("top", = e.pageY-elH).css("position", "absolute");=0A= }=0A= }=0A= }=0A= =0A= function hideRef(e) {=0A= if($("div#hovering-ref").length) {=0A= $("div#hovering-ref").remove();=0A= }=0A= }=0A= =0A= function getHWCiting() {=0A= var citingA =3D $("#cb-hw-citing-articles");=0A= if (citingA.length) {=0A= var newA =3D 'Loading citing = article data...';=0A= citingA.replaceWith(newA);=0A= var href =3D citingA.attr("href");=0A= var id =3D '';=0A= if (href && (href.indexOf('?') > 0)) {=0A= var args =3D href.substring(href.indexOf('?') + 1).split('&');=0A= for (var i =3D 0; i < args.length; i++) {=0A= if (args[i].toLowerCase().indexOf('legid=3D') =3D=3D 0) {=0A= id =3D args[i].substring(args[i].indexOf('=3D') + 1);=0A= if (id.indexOf('#') > 0) {=0A= id =3D id.substring(0, id.indexOf('#'));=0A= }=0A= }=0A= }=0A= if (!(id =3D=3D '')) {=0A= var host =3D document.location.protocol + "//" + = document.location.host;=0A= var ajaxUrl =3D host + '/cited-by/' + id.replace(/;/,'/');=0A= $.ajax({=0A= url: ajaxUrl,=0A= dataType: "html",=0A= type: "GET",=0A= error: ajaxErr,=0A= success: addHWCiting,=0A= complete: ajaxComplete=0A= });=0A= =0A= }=0A= }=0A= }=0A= }=0A= function getHWRelatedURLs() {=0A= var relatedURLsA =3D $("#cb-related-urls");=0A= var relatedURLsMsg =3D = getSiteOption('relatedWebPageLoadingText','Loading related web page = data...'); =0A= if (relatedURLsA.length) {=0A= var newA =3D '' + = relatedURLsMsg + '';=0A= relatedURLsA.replaceWith(newA);=0A= var href =3D relatedURLsA.attr("href");=0A= var id =3D '';=0A= if (href && (href.indexOf('?') > 0)) {=0A= var args =3D href.substring(href.indexOf('?') + 1).split('&');=0A= for (var i =3D 0; i < args.length; i++) {=0A= if (args[i].toLowerCase().indexOf('legid=3D') =3D=3D 0) {=0A= id =3D args[i].substring(args[i].indexOf('=3D') + 1);=0A= if (id.indexOf('#') > 0) {=0A= id =3D id.substring(0, id.indexOf('#'));=0A= }=0A= }=0A= }=0A= if (!(id =3D=3D '')) {=0A= var host =3D document.location.protocol + "//" + = document.location.host;=0A= var ajaxUrl =3D host + '/related-web-pages/' + id.replace(/;/,'/');=0A= $.ajax({=0A= url: ajaxUrl,=0A= dataType: "html",=0A= type: "GET",=0A= error: ajaxErr,=0A= success: addRelatedURLs,=0A= complete: ajaxComplete=0A= });=0A= =0A= }=0A= }=0A= }=0A= }=0A= function getPatientInformData() {=0A= var pInformA =3D $("#cb-patientinform");=0A= if (pInformA.length) {=0A= var newA =3D 'Loading = patientINFORMation...';=0A= pInformA.replaceWith(newA);=0A= var href =3D pInformA.attr("href");=0A= var id =3D '';=0A= if (href && (href.indexOf('?') > 0)) {=0A= var args =3D href.substring(href.indexOf('?') + 1).split('&');=0A= for (var i =3D 0; i < args.length; i++) {=0A= if (args[i].toLowerCase().indexOf('legid=3D') =3D=3D 0) {=0A= id =3D args[i].substring(args[i].indexOf('=3D') + 1);=0A= if (id.indexOf('#') > 0) {=0A= id =3D id.substring(0, id.indexOf('#'));=0A= }=0A= }=0A= }=0A= if (!(id =3D=3D '')) {=0A= var host =3D document.location.protocol + "//" + = document.location.host;=0A= var ajaxUrl =3D host + '/related-web-pages/patientinform/' + = id.replace(/;/,'/');=0A= $.ajax({=0A= url: ajaxUrl,=0A= dataType: "html",=0A= type: "GET",=0A= error: ajaxErr,=0A= success: addPatientInform,=0A= complete: ajaxComplete=0A= });=0A= =0A= }=0A= }=0A= }=0A= }=0A= function getISIRelated() {=0A= var relatedA =3D $("#cb-isi-similar-articles");=0A= if (relatedA.length) {=0A= var newA =3D 'Loading Web = of Science article data...';=0A= relatedA.replaceWith(newA);=0A= var href =3D relatedA.attr("href");=0A= var id =3D '';=0A= if (href) {=0A= var hrefDec =3D decodeURI(href);=0A= if ((hrefDec.indexOf('?') > 0)) {=0A= var args =3D hrefDec.substring(hrefDec.indexOf('?') + 1).split('&');=0A= for (var i =3D 0; i < args.length; i++) {=0A= var argDec =3D decodeURIComponent(args[i]);=0A= if (argDec.toLowerCase().indexOf('access_num=3D') =3D=3D 0) {=0A= id =3D argDec.substring(argDec.indexOf('=3D') + 1);=0A= if (id.indexOf('#') > 0) {=0A= id =3D id.substring(0, id.indexOf('#'));=0A= }=0A= }=0A= }=0A= if (!(id =3D=3D '')) {=0A= var host =3D document.location.protocol + "//" + = document.location.host;=0A= var ajaxUrl =3D host + '/isi-links/has-related/' + = id.replace(/;/,'/');=0A= $.ajax({=0A= url: ajaxUrl,=0A= dataType: "html",=0A= type: "GET",=0A= error: ajaxErr,=0A= success: addISIRelated,=0A= complete: ajaxComplete=0A= });=0A= }=0A= }=0A= }=0A= }=0A= }=0A= function getCiting(service, msg, pathseg, successFn) {=0A= var citingA =3D $("#cb-" + service + "-citing-articles");=0A= if (citingA.length) {=0A= var newA =3D '' = + msg + '';=0A= citingA.replaceWith(newA);=0A= var href =3D citingA.attr("href");=0A= var id =3D '';=0A= if (href) {=0A= var hrefDec =3D decodeURI(href);=0A= if ((hrefDec.indexOf('?') > 0)) {=0A= var args =3D hrefDec.substring(hrefDec.indexOf('?') + 1).split('&');=0A= for (var i =3D 0; i < args.length; i++) {=0A= var argDec =3D decodeURIComponent(args[i]);=0A= if (argDec.toLowerCase().indexOf('access_num=3D') =3D=3D 0) {=0A= id =3D argDec.substring(argDec.indexOf('=3D') + 1);=0A= if (id.indexOf('#') > 0) {=0A= id =3D id.substring(0, id.indexOf('#'));=0A= }=0A= }=0A= }=0A= if (!(id =3D=3D '')) {=0A= var host =3D document.location.protocol + "//" + = document.location.host;=0A= var ajaxUrl =3D host + '/' + service + '-links/' + pathseg + = id.replace(/;/,'/');=0A= $.ajax({=0A= url: ajaxUrl,=0A= dataType: "html",=0A= type: "GET",=0A= error: ajaxErr,=0A= success: successFn,=0A= complete: ajaxComplete=0A= });=0A= }=0A= }=0A= }=0A= }=0A= }=0A= =0A= function getISICiting() {=0A= var citingA =3D $("#cb-isi-citing-articles");=0A= if (citingA.length) {=0A= var newA =3D 'Loading Web of = Science citing article data...';=0A= citingA.replaceWith(newA);=0A= var href =3D citingA.attr("href");=0A= var id =3D '';=0A= if (href) {=0A= var hrefDec =3D decodeURI(href);=0A= if ((hrefDec.indexOf('?') > 0)) {=0A= var args =3D hrefDec.substring(hrefDec.indexOf('?') + 1).split('&');=0A= for (var i =3D 0; i < args.length; i++) {=0A= var argDec =3D decodeURIComponent(args[i]);=0A= if (argDec.toLowerCase().indexOf('access_num=3D') =3D=3D 0) {=0A= id =3D argDec.substring(argDec.indexOf('=3D') + 1);=0A= if (id.indexOf('#') > 0) {=0A= id =3D id.substring(0, id.indexOf('#'));=0A= }=0A= }=0A= }=0A= if (!(id =3D=3D '')) {=0A= var host =3D document.location.protocol + "//" + = document.location.host;=0A= var ajaxUrl =3D host + '/isi-links/' + id.replace(/;/,'/');=0A= $.ajax({=0A= url: ajaxUrl,=0A= dataType: "html",=0A= type: "GET",=0A= error: ajaxErr,=0A= success: addISICiting,=0A= complete: ajaxComplete=0A= });=0A= }=0A= }=0A= }=0A= }=0A= }=0A= function getEntrezLinks() {=0A= var entrezDiv =3D $("#cb-entrez-links-placeholder");=0A= if (entrezDiv.length) {=0A= var entrezA =3D entrezDiv.children("a");=0A= if (entrezA) {=0A= var host =3D document.location.protocol + "//" + = document.location.host;=0A= var ajaxUrl =3D host + entrezA.attr("href");=0A= $.ajax({=0A= url: ajaxUrl,=0A= dataType: "html",=0A= type: "GET",=0A= error: ajaxErr,=0A= success: addEntrezLinks,=0A= complete: ajaxComplete=0A= });=0A= }=0A= }=0A= }=0A= =0A= function ajaxErr(req, msg, e) {=0A= }=0A= function ajaxComplete(req, msg) {=0A= }=0A= function updateCBItem(cbItem, newHTML, hasData) {=0A= var parentItem =3D cbItem.parents("li").eq(0);=0A= cbItem.replaceWith(newHTML);=0A= if (!hasData) {=0A= // hide the parent li=0A= if (parentItem.length) {=0A= modClass(parentItem,"nodata","");=0A= // check if there are any siblings still being displayed=0A= var otherItems =3D parentItem.siblings();=0A= var allItemsEmpty;=0A= if (otherItems.length) {=0A= if (otherItems.length =3D=3D otherItems.filter(".nodata").length) {=0A= allItemsEmpty =3D true=0A= }=0A= else {=0A= allItemsEmpty =3D false;=0A= }=0A= }=0A= else {=0A= allItemsEmpty =3D true;=0A= }=0A= if (allItemsEmpty) {=0A= var cbsection =3D parentItem.parents("div.cb-section").eq(0);=0A= if (cbsection.length) {=0A= modClass(cbsection,"nodata","");=0A= }=0A= }=0A= }=0A= }=0A= // in frameset fix targets on child links, forms=0A= fixFrameLinks(parentItem.find("a,form"));=0A= fixColHeights(2);=0A= }=0A= function fixFrameLinks(jqItems) {=0A= if ((gIsFrameset !=3D null) && gIsFrameset) {=0A= jqItems.each(=0A= function(i) {=0A= var href =3D $(this).attr("href");=0A= var action =3D $(this).attr("action"); // if form=0A= if ((href !=3D null) || (action !=3D null)) {=0A= var inFrameAnchor =3D ((href !=3D null) && = (((/frameset=3D/.test(href)) && (/#/.test(href))) || = (href.substring(0,1) =3D=3D '#')));=0A= if ((!inFrameAnchor) || (action !=3D null)) {=0A= $(this).attr("target", "_top");=0A= }=0A= }=0A= }=0A= );=0A= }=0A= }=0A= function addRelatedURLs(xhtmlData) {=0A= var cbA =3D $("#cb-loading-related-urls");=0A= if (gIsFrameset) {=0A= if (cbA.length) {=0A= updateCBItem(cbA, '', false);=0A= }=0A= }=0A= else if (xhtmlData && !(xhtmlData.indexOf('=3D 0)) {=0A= $("#related-urls").replaceWith(xhtmlData);=0A= var relatedWebPagesLabel =3D getSiteOption('relatedWebPagesLabel', = 'Related Web Pages');=0A= fixColHeights(1);=0A= if (cbA.length) {=0A= updateCBItem(cbA, '' + relatedWebPagesLabel = + '', true);=0A= }=0A= }=0A= else {=0A= if (cbA.length) {=0A= updateCBItem(cbA, '', false);=0A= }=0A= }=0A= }=0A= function addPatientInform(xhtmlData) {=0A= var cbA =3D $("#cb-loading-patientinform");=0A= if (gIsFrameset) {=0A= if (cbA.length) {=0A= updateCBItem(cbA, '
        Not = available in this view
        ', false);=0A= }=0A= }=0A= else if (xhtmlData && !(xhtmlData.indexOf('=3D 0)) {=0A= $("#patientinform-links").replaceWith(xhtmlData);=0A= fixColHeights(1);=0A= if (cbA.length) {=0A= updateCBItem(cbA, 'patientINFORMation', true);=0A= }=0A= }=0A= else {=0A= if (cbA.length) {=0A= updateCBItem(cbA, '
        No = patientINFORMation available for this article
        ', false);=0A= }=0A= }=0A= }=0A= function addHWCiting(xhtmlData) {=0A= var cbA =3D $("#cb-loading-hw-cited");=0A= if (gIsFrameset) {=0A= if (cbA.length) {=0A= updateCBItem(cbA, '
        Not available = in this view
        ', false);=0A= }=0A= }=0A= else if (xhtmlData) {=0A= $("#content-block").append(xhtmlData);=0A= var hwCitingLabel =3D getSiteOption('hwCitingLabel', 'View citing = article information');=0A= fixColHeights(1);=0A= if (cbA.length) {=0A= updateCBItem(cbA, '' + hwCitingLabel + '', = true);=0A= }=0A= }=0A= else {=0A= if (cbA.length) {=0A= updateCBItem(cbA, '
        No citing = articles
        ', false);=0A= }=0A= }=0A= }=0A= function addISIRelated(xhtmlData) {=0A= var cbA =3D $("#cb-isi-similar-articles");=0A= if (xhtmlData && !(xhtmlData.indexOf('=3D 0)) {=0A= if (cbA.length) {=0A= updateCBItem(cbA, xhtmlData, true);=0A= }=0A= }=0A= else {=0A= if (cbA.length) {=0A= updateCBItem(cbA, '', false);=0A= }=0A= }=0A= }=0A= function addISICiting(xhtmlData) {=0A= var cbA =3D $("#cb-loading-isi-cited");=0A= if (xhtmlData && !(xhtmlData.indexOf('=3D 0)) {=0A= if (cbA.length) {=0A= updateCBItem(cbA, xhtmlData, true);=0A= }=0A= }=0A= else {=0A= if (cbA.length) {=0A= updateCBItem(cbA, '
        No Web of = Science citing articles
        ', false);=0A= }=0A= }=0A= }=0A= function addScopusCiting(xhtmlData) {=0A= var cbA =3D $("#cb-loading-scopus-cited");=0A= if (xhtmlData && (xhtmlData.indexOf('=3D 0)) {=0A= if (cbA.length) {=0A= updateCBItem(cbA, xhtmlData, true);=0A= }=0A= }=0A= else {=0A= if (cbA.length) {=0A= updateCBItem(cbA, '
        No Scopus = citing articles
        ', false);=0A= }=0A= }=0A= }=0A= function addEntrezLinks(xhtmlData) {=0A= var entrezDiv =3D $("#cb-entrez-links-placeholder");=0A= =0A= if (xhtmlData && (xhtmlData.indexOf('        =3D 0)) {=0A= if (entrezDiv.length) {=0A= updateCBItem(entrezDiv, xhtmlData, true);=0A= }=0A= }=0A= else {=0A= if (entrezDiv.length) {=0A= updateCBItem(entrezDiv, '', false);=0A= }=0A= }=0A= }=0A= =0A= function updateSBLinks() {=0A= var fbLink =3D $("a.sb-facebook");=0A= if (fbLink.length) {=0A= fbLink.click(=0A= function(e) {=0A= window.open(this.href, 'sharer', = 'toolbar=3D0,status=3D0,width=3D626,height=3D436');=0A= e.preventDefault();=0A= }=0A= );=0A= }=0A= }=0A= =0A= function addDockedNav() {=0A= var slugEl =3D $("#col-2 #slugline");=0A= // get direct children li elements only=0A= var artViews =3D $("#article-cb-main .cb-section ol:has(li = a[rel^=3D'view-']) > li").clone();=0A= var newDiv =3D '
        ';=0A= $("#col-2").append(newDiv);=0A= $("#col-2 #docked-nav").hide();=0A= var newDivJQuery =3D $("#docked-nav");=0A= newDivJQuery.append('

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