From: Subject: Gene Expression Microarray Analysis Reveals YKL-40 to Be a Potential Serum Marker for Malignant Character in Human Glioma -- Tanwar et al. 62 (15): 4364 -- Cancer Research Date: Sat, 9 Feb 2008 10:21:40 +0100 MIME-Version: 1.0 Content-Type: multipart/related; type="text/html"; boundary="----=_NextPart_000_003B_01C86B05.8F4FA870" X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3198 This is a multi-part message in MIME format. ------=_NextPart_000_003B_01C86B05.8F4FA870 Content-Type: text/html; charset="Windows-1252" Content-Transfer-Encoding: quoted-printable Content-Location: http://cancerres.aacrjournals.org/cgi/content/full/62/15/4364 Gene Expression Microarray Analysis Reveals YKL-40 to = Be a Potential Serum Marker for Malignant Character in Human Glioma -- = Tanwar et al. 62 (15): 4364 -- Cancer Research 3D"Cancer 3D"
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[Cancer Research 62, 4364-4368, August 1, = 2002]
=A9 2002 American = Association=20 for Cancer Research

Clinical Investigations

Gene Expression Microarray Analysis Reveals YKL-40 to Be a Potential = Serum=20 Marker for Malignant Character in Human Glioma1=20

Meena K. Tanwar, Mark R. = Gilbert=20 and Eric C. Holland2=20

Departments of Surgery (Neurosurgery), Neurology, and = Cell=20 Biology, Memorial Sloan-Kettering Cancer Center, New York, New York = 10021 [M. K.=20 T., E. C. H.], and Department of Neuro-Oncology, University of Texas, = M.D.=20 Anderson Cancer Center, Houston, Texas 77030 [M. R. G.]


3D"   =20 ABSTRACT
Top
3D"ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
 
W= e have=20 identified a potential serum marker for glioblastoma = multiforme (GBM)=20 usingmicroarray analysis from samples of GBM. We compared the = gene=20 expression profile of 19 gliomas to pooled normal brain by=20 competitive hybridization of RNA from each tumor sample to a = pooled=20 sample of RNA isolated from normal brain tissue using the = Incyte=20 10,000 gene expression array. The most differentially = expressed gene=20 in this analysis encodes a secreted glycoprotein and is = referred to=20 as YKL-40. YKL-40 mRNA was detected in the GBM = samples=20 with a range of 3- to 62-fold elevation over normal brain. It = has=20 been reported previously that this protein is expressed in = pathologic=20 conditions of extracellular matrix degradation and = angiogenesis, such=20 as rheumatoid arthritis, severe osteoarthritis, hepatic = fibrosis,=20 primary colorectal cancer, and metastatic breast cancer. = These data=20 suggest YKL-40 may be involved in extracellular matrix = degradation=20 and/or angiogenesis. Western blot analysis of glioma samples = for=20 YKL-40 protein levels revealed substantial elevation in 3D~=2065% of = GBMs and=20 undetectable levels in lower-grade gliomas (grade II and III) = or=20 normal brain tissue. We performed ELISA analysis on serum = samples of=20 glioma patients to determine whether this protein would = correlate=20 with the presence of tumor and tumor grade or burden. YKL-40=20 serum levels were substantially elevated in many of the GBM=20 patients. Statistical analysis of these data indicates that = in=20 patients with glioma, serum YKL-40 levels correlate with = tumor=20 grade and potentially tumor burden in GBM.


3D"   =20 INTRODUCTION
Top
ABSTRACT
3D"INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
 
G= BM3=20 is the most common and most malignant primary brain tumor in = adults=20 (1)=20 . Primary GBMs arise de novo with no evidence of = previous=20 lower-grade glioma. Secondary GBMs arise from pre-existing = anaplastic=20 astrocytomas, grade III glioma, or diffuse astrocytoma, grade = II. Of=20 the gliomas, grade IV GBM has the worst prognosis with a = median=20 survival of 3D~12 months, even after surgical resection, = radiation therapy,=20 and chemotherapy. Patients with anaplastic astrocytomas have = an=20 average survival of 3 years, whereas patients with grade II = gliomas=20 have the best prognosis, with >5 years median survival (2)=20 . Histologically, GBMs are characterized by high mitotic = activity,=20 nuclear atypia, microvascular proliferation, and necrosis. = However,=20 in the future, molecular subgrouping of the gliomas could aid = in the=20 classification of these tumors and may eventually help to = better=20 predict survival (3)=20 .

Several distinct prognostic subgroups exist within each of these = clinical grades, and consequently the survival of glioma = patients=20 can be quite variable (4=20 ,=20 5) . Currently, the determination of treatment response = and tumor=20 progression is monitored using imaging techniques such as = molecular=20 resonance imaging or computed tomography. Often extensive = tumor=20 progression occurs in the interval between different imaging = studies=20 and/or goes undetected from lack of contrast enhancement. An=20 alternative measure or marker of tumor burden may potentially = permit=20 early detection of treatment failure and allow for more rapid = changes=20 in therapeutic strategy.

Development of serum markers that are specific for a particular=20 tumor type (6)=20 or that correlate with biological properties of the tumor = could be=20 used for diagnosis and as a measure of response to treatment, = and=20 would greatly enhance patient care. If serum markers were = identified=20 that correlate with the activity of certain signaling = pathways within=20 the tumor, patients could be directed to clinical trials = using=20 specific agents aimed at that particular tumor biology. With = such a=20 set of markers that could predict and monitor response to = therapy,=20 patients would be better advised of the likely benefits of = aggressive=20 treatment. One method for identification of candidate markers = is=20 using gene expression microarray analysis (7=20 ,=20 8) . Although many studies have been done on gene = expression=20 profiling of specific tumor types, and differentially = expressed genes=20 in these tumors have been reported, few of these studies have = resulted in clinical application.

To identify potential serum markers as genes overexpressed in = GBMs=20 relative to lower-grade gliomas and normal brain, we performed = cDNA=20 microarray gene expression profiling of a series of GBMs and=20 lower-grade gliomas. In this analysis, we found that the most = differentially expressed gene in the series of GBMs versus=20 normal brain was YKL-40. YKL-40 encodes a secreted = protein=20 with sequence similarity to glycosyl hydrolases (9)=20 and has been identified previously as being expressed in = other cancer=20 types such as breast and colon (10=20 ,=20 11) . Little to no differential expression of = YKL-40 was=20 seen in lower-grade tumors relative to normal brain. The = expression=20 of YKL-40 protein, confirmed by Western blot analysis of = tumor=20 extracts, was also specific to GBMs and undetectable in = lower-grade=20 gliomas and normal brain. To ascertain whether YKL-40 could = be an=20 accurate marker for GBM presence and tumor burden, we = measured YKL-40=20 serum levels in patients diagnosed with various types and = grades of=20 glioma, using a YKL-40-specific ELISA assay. Our data = indicate a=20 significant difference in YKL-40 serum concentrations between = normal=20 subjects and GBM patients, as well as between lower-grade = glioma=20 and GBM patients. Furthermore, there was a trend toward high=20 YKL-40 levels and high tumor burden, although this did not = reach=20 statistical significance. These preliminary data suggest that = YKL-40=20 levels may be an efficient and accurate indicator of glioma=20 presence and grade.


3D"   =20 MATERIALS AND METHODS
Top
ABSTRACT
INTRODUCTION
3D"MATERIALS AND = METHODS
RESULTS
DISCUSSION
REFERENCES
 
<= STRONG>Tissue=20 Collection.
Patient tumor and normal brain samples were = collected=20 from patients at M.D. Anderson Cancer Center at the time of = surgical=20 resection. Normal brain sample controls were obtained as = overlying=20 cortex during routine resection of deep intercerebral = metastases.=20 Specimens were snap frozen in liquid nitrogen and stored at=20 -80=B0C until use. All of the patients signed consent for=20 collection and analysis of samples.

Tissue Preparation and Microarray Analysis.
For = RNA and=20 protein extraction, the tissue was ground to powder using a = mortar=20 and pestle in the presence of liquid nitrogen. Nine grade IV = GBMs, 5=20 grade III gliomas, and 5 grade II gliomas were used in the = array=20 analysis. RNA was isolated from 100 to 400 mg of tissue per = sample.=20 Standard TRIzol preparation protocol (Life Technologies, = Inc.,=20 Rockville, MD) and reagents were used for total RNA = isolation. To=20 isolate polyadenylated RNA, the total RNA was twice passed = over an=20 Oligotex mRNA isolation column (Qiagen, Valencia, CA) = following the=20 manufacturer=92s protocol. The RNA samples were quantified = using a=20 spectrophotometer and visualized on a Tris-borate EDTA gel = for=20 quality assurance. The mRNA samples were converted to cDNA = and=20 subsequently hybridized to the Human V cDNA microarray by = Incyte=20 Genomics (St. Louis, MO). The data were analyzed using GEM = Tools 2.5=20 software.

Protein Extraction.
Sample collection and = preparation was=20 performed as described above. Protein extraction was = performed using=20 lysis buffer [100 mM NaCl, 30 mM Tris-HCl (pH 7.6), 1% NP40, 30 mM NaF, 1=20 mM sodium orthovanadate, 0.5 mM phenylmethylsulfonyl fluoride, protease = inhibitor=20 mixture tablets (Roche, Indianapolis, IN)], and the tissue = was=20 homogenized on ice using a sterile, plastic disposable = pestle. The=20 samples were allowed to incubate on ice for 30 min; then = centrifuged=20 at 14,000 x g for 10 min at = 4=B0C. The=20 supernatant was transferred to a fresh microfuge tube, and = protein=20 quantitation was performed using the BCA Protein Assay kit = (Pierce,=20 Rockford, IL) and a standard spectrophotometer.

Western Blot Analysis.
Fifty micrograms of each = protein=20 sample was separated on an SDS-PAGE gel. The protein samples = were=20 transferred overnight to nitrocellulose membrane (Osmonics=20 Fischer-Scientific, Pittsburgh, PA). For protein detection, = the=20 membrane was incubated in 5% milk in TBS-T for 1 h at room=20 temperature. The membrane was then incubated for 1 h with 4 = mg of=20 YKL-40 polyclonal antibody (Metra Biosystems, Mountain View, = CA) in=20 5% dried milk-TBS-T. The membrane was then washed three times = in=20 TBS-T, 5 min each, and incubated with the secondary = antirabbit=20 antibody, 1:2000 dilution (Roche) for 1 h. Again the membrane = was=20 washed three times in TBS-T, and antibody-bound protein was = detected=20 by adding enhanced chemiluminescence reagent (Amersham = Pharmacia=20 Biotechnology, Piscataway, NJ) for 1 min and exposing the = membrane to=20 Kodak film.

Blood Collection and Serum Separation.
All of the = blood=20 samples were collected from patients at M.D. Anderson Cancer = Center=20 diagnosed with glioma through collaboration with the = Neuro-Oncology=20 Clinic. Normal blood samples were collected from volunteer = subjects=20 with no known malignancies. The blood samples were allowed to = clot at=20 room temperature for no more than 72 h, and were then = centrifuged at=20 4=B0C for 5 min at 1000 rpm. The serum (upper phase) was = aliquoted and=20 stored at -20=B0C until use.

YKL-40 ELISA Assay.
YKL-40 levels were = determined, in=20 duplicate, for all of the serum samples, using the YKL-40 = ELISA kit=20 from Metra Biosystems according to the manufacturer=92s = protocol.=20 Protein concentrations were determined as absorbances using = the=20 Bio-Rad Benchmark Microplate Reader.

Statistical Analysis.
The histological grade of = each=20 glioma according to the WHO criteria, as well as the patient = disease=20 status was determined for each sample collected. Disease = status was=20 stratified into a high tumor burden group and a low tumor = burden=20 group. Patients with newly diagnosed tumors awaiting initial = surgery=20 or who underwent only stereotactic biopsy were considered to = have a=20 high tumor burden. Patients with radiographic evidence of = recurrent=20 tumor growth were also included in this group. Patients who=20 underwent a gross total or near total resection, had no = active=20 disease, or had evidence of response to treatment were placed = in=20 the low tumor burden group. Statistical analysis of data in = each=20 group was done using Student=92s t test.


3D"   =20 RESULTS
Top
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
3D"RESULTS
DISCUSSION
REFERENCES
 
<= STRONG>Experimental=20 Design.
We were interested in studying gene expression = changes=20 that occur in malignant and nonmalignant gliomas compared = with=20 normal brain tissue to find potential clinical markers for = this=20 disease. Our approach was to start with a microarray analysis = using=20 the Incyte Genomics Human V cDNA microarray. Tumor mRNA = samples=20 were labeled with cy-5 and hybridized against the = cy-3-labeled=20 normal mRNA control. For this purpose, we chose to pool = samples=20 of normal brain mRNA, generating a constant control to be = used=20 on each chip analyzed (Fig. 1) . The normal brain samples were obtained from = patients=20 with deep metastatic brain tumors, cases in which removal of=20 overlying normal cortex was required for safe resection of = the=20 metastatic tumor. mRNA was isolated from normal frontal, = temporal,=20 parietal, and occipital cortex samples, and combined to make = one=20 common control. To preserve the quality of the mRNA during = tissue=20 collection, all of the specimens were snap frozen in liquid = nitrogen=20 immediately after removal from the patient during surgery and = subsequently stored at -80=B0C.



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  		Fig. 1. Microarray strategy. All = of the=20 glioma mRNA samples were hybridized against RNA from pooled = normal=20 brain tissue obtained during resection of deep metastases. = This=20 strategy allows comparison of gene expression profiles = between each=20 tumor and normal brain as well as between tumors. NB, = normal=20 brain; G, GBM; L, lower-grade glioma. =

 = ;
Because=20 each tumor mRNA sample was normalized against a pooled = control, we=20 were able to compare gene expression changes across different = tumors.=20 The differential expression patterns obtained were the = comparative=20 value of gene expression in a given tumor versus = normal brain=20 and represented an elevation or depression of gene expression = in=20 tumor tissue over normal tissue. For a given array = hybridization,=20 >97% of genes analyzed fell within a 2-fold variance = between tumor=20 and normal brain (Fig. 2A) .



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  		Fig. 2. Microarray analysis of = glioma=20 versus normal brain. A, gene expression display of = results=20 from a given array hybridization. Shows that >95% of all = genes=20 were differentially expressed by < 2-fold difference. = B,=20 graph illustrating YKL-40 tumor mRNA levels as fold increase = relative to normal brain. Expression elevation was not seen = in=20 lower-grade gliomas (LGR) versus normal = control.=20 C, comparison of differential mRNA levels encoding = YKL-40=20 with transcripts encoding proteins involved in = endoproteolytic and=20 endoglycosidase degradation of the ECM. YKL-40 mRNA is = significantly=20 overexpressed compared with all endoproteases on the array. = Order of=20 genes graphed beginning from the back: YKL-40, = Chitobiase, MMP-16=20 (matrix metalloproteinase-16), MMP-14, MMP-12, MMP-11, = MMP-7, MMP-2,=20 TIMP-1 (tissue inhibitor of metalloproteinase 1), TIMP-3, = urokinase=20 receptor plasminogen activator, tissue plasminogen = activator, PAI-II=20 (plasminogen activator inhibitor-II), PAI-I, and=20 plasminogen. Numerical values for differential = expression=20 levels displayed in this graph are given in Table 1 . D, Western blot analysis of glioma = and normal=20 brain samples for YKL-40 protein showing that it is = significantly=20 elevated in 50% of the GBMs and undetectable in lower-grade = gliomas=20 (LGR) as well as a representative normal brain sample = (N). =

 = ;
YKL-40=20 Is Overexpressed in a Subset of GBMs.
There were a number of = genes=20 significantly up-regulated in the GBM samples but not in the=20 lower-grade tumors, compared with the normal control. Of = these genes,=20 the most highly elevated gene expression was seen for=20 chitinase-3-like-1, also known as human cartilage=20 glycoprotein-39 and YKL-40 (Fig. 2B) . YKL-40 mRNA has also been shown to = be elevated=20 in GBM samples by serial analysis of gene expression analysis = (12)=20 and by oligonucleotide array analysis (13)=20 . These independent methods of determining gene expression = levels=20 additionally validate our findings.

Review of the literature reveals that the gene for YKL-40 is=20 located on chromosome 1q32 and is a mammalian member of = family=20 18 glycosyl hydrolases, a gene family that includes bacterial = and fungal chitinases (9=20 ,=20 14) . YKL-40 has significant sequence similarity to the=20 chitin-degrading enzyme, chitinase; it has been shown to bind = chitin=20 but retains no chitinase activity and has not been determined = to have=20 any other enzymatic activity or function (15)=20 . The full spectrum of mammalian polysaccharide structures = that bind=20 to YKL-40 and the function of this protein in human glioma = biology=20 are unknown. YKL-40 is secreted by chondrocytes, synovial = cells,=20 neutrophils, and the osteosarcoma cell line MG-63 (9=20 ,=20 16 ,=20 17) .

Several lines of circumstantial evidence indicate that YKL-40 = may=20 promote degradation of the ECM or angiogenesis. YKL-40 is not = present=20 in normal adult cartilage but is detectable in human = arthritic=20 cartilage and during periods of cartilage generation, = remodeling, and=20 degradation. For example, patients with rheumatoid arthritis = have=20 high levels of serum-detectable YKL-40 (18)=20 . In addition, elevated YKL-40 levels have been detected in=20 patients with hepatic fibrosis and cirrhosis (19)=20 , and in bovine breast tissue during mammary gland involution = after=20 cessation of lactation (20)=20 , processes that involve tissue destruction and ECM remodeling. =

Because of the potential role of YKL-40 in ECM degradation, = we=20 compared its differential expression to that of other proteins=20 involved in this process such as endoproteases and = endoglycosidases=20 present on the microarray, many of which have been shown to = be=20 important factors in glioma biology (Fig. 2C) . Although some of these genes demonstrated = elevated=20 mRNA levels in the tumor samples, when graphed together with=20 YKL-40, their relative differential expression levels = were=20 barely detectable because of the substantial elevation of=20 YKL-40 mRNA in the GBM samples. Differential = expression values=20 of YKL-40, endoglycosidases, endoproteases, and other = ECM=20 components on the array are shown in Table 1 .


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  		Table 1 Differential gene = expression=20 values of ECM-related genes =

 = ;
To=20 determine whether the elevation in YKL-40 mRNA levels = correlated=20 with protein production, we analyzed protein extract from = human=20 GBMs, lower-grade gliomas, and normal brain tissue by Western = blot analysis. We found that YKL-40 protein was dramatically=20 elevated in a subset of the GBMs, and essentially below the=20 detection limit for Western blot analysis, in both = lower-grade=20 tumors and normal brain extract (Fig. 2D) .

YKL-40 Levels Are Stable in Collected Serum.
In a = recent=20 publication, YKL-40 serum values were found to increase = significantly=20 over time when blood samples (before serum collection) were = stored at=20 room temperature for >24 h or at 4=B0C for >72 h before = serum=20 isolation (21)=20 . To determine whether our serum collection protocol = introduced=20 variable measurements because of collection techniques, we = performed=20 a time course experiment. We collected two vials of blood = from each=20 of two normal, healthy subjects. The blood samples were=20 immediately centrifuged, and 150 =B5l of serum was collected = from=20 each vial and stored at -80=B0C. Next, one vial from each = subject=20 was kept at room temperature, and the other was kept at = 4=B0C.=20 Serum was collected from each vial at time points of 6 h, 24 = h,=20 72 h, and 7 days, stored at -80=B0C, and subsequently analyzed = by ELISA=20 as described in "Materials and Methods." We found that YKL-40 = levels=20 did not show any significant increase or decrease through = continuous=20 storage at either room temperature or at 4=B0C (Fig. = 3A) .



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  		Fig. 3. YKL-40 in the serum of = glioma=20 patients and normal controls, and as a function of tumor = burden.=20 A, time course of YKL-40 levels in serum from two = normal=20 volunteers. YKL-40 levels appear stable in serum stored at = room=20 temperature (3D""=203D""=203D""=20)=20 and at 4=B0C (3D""=20=20 3D"" 3D""=20)=20 over time. B, categorical illustration of YKL-40 = serum levels=20 in normal controls, lower-grade gliomas, and GBMs. = *=20 designates the mean YKL-40 value of each subgroup. C, = YKL-40=20 levels displayed as a measure of tumor burden in patients = with grade=20 IV GBM and compared with normal controls. Tumor burden was = defined,=20 at the time of serum collection, as presence of tumor based = on pre-=20 or postoperative patient status. =

 = ;
YKL-40=20 Levels in the Serum of Patients Correlate with Glioma = Grade.
On the=20 basis of the above data, YKL-40 appears to be a promising = candidate=20 serum marker for a subset of GBMs. To test whether YKL-40 = could be=20 used as a clinical marker correlating with glioma grade and = tumor=20 burden, we collected serum samples from patients diagnosed = with=20 gliomas of various grades and 14 normal healthy subjects. = Serum=20 levels of YKL-40 were measured in 45 patients with GBM (grade = IV), 20=20 patients with lower-grade glioma (grades II and III), and 14 = controls=20 (Fig. 3B) . The grade II and III gliomas were grouped = together=20 for this analysis based on the results of YKL-40 gene=20 expression described above, which indicated that both = histological=20 grades had low expression. Comparisons of serum YKL-40 levels = were=20 made between glioblastoma and controls, glioblastoma and = lower-grade=20 gliomas, lower-grade gliomas and controls, and high = versus low=20 tumor burden in patients with glioblastoma. The results of = these=20 comparisons are reported in Table 2 . Statistically significant differences were = detected=20 between YKL-40 serum levels in patients with glioblastoma = compared=20 with controls (P < 0.001) and compared with patients = with=20 lower-grade gliomas (P =3D 0.037), as well as between=20 lower-grade tumors compared with controls (P < = 0.001). No=20 significant differences were detected between high = versus low=20 tumor burden in serum YKL-40 from patients with GBM. The lack = of=20 statistical significance in the tumor burden analysis is = because of a=20 single outlier in the low tumor burden group, illustrated in = Fig.=20 3C ; however, there is no clinical explanation = for the=20 high serum YKL-40 levels in this patient.


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  		Table 2 Statistical analysis of = serum=20 YKL-40 levels =

 = ;

3D"   =20 DISCUSSION
Top
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
3D"DISCUSSION
REFERENCES
 
L= evels of=20 YKL-40 protein can be reproducibly measured in the serum of = patients=20 with malignant primary brain tumors, reflecting the fact that = YKL-40=20 is a secreted glycoprotein. Our data suggest low but = detectable=20 YKL-40 in serum of most normal controls and that = significantly higher=20 levels are detected in serum from patients with GBM compared = with the=20 lower-grade gliomas. The origin of YKL-40 production in = normal=20 individuals is not known. Consistent with the gene expression = and=20 Western blot data, not all of the high-grade gliomas = manifested=20 elevated serum levels of the YKL-40 protein. Furthermore, in = some=20 cases, the diagnosis attributed to a patient was made = significantly=20 earlier and may not have reflected the status of the tumor at = the=20 time of blood draw. This contributed to the large SD for = YKL-40=20 levels in GBM samples. Because not all of the patients had = molecular=20 resonance imaging scans done at the time of blood withdrawal, = it was=20 not possible to correlate radiographic tumor volume to YKL-40 = levels. Therefore, we categorized the patients into low and = high=20 tumor burden based on clinical grounds such as pre- = versus=20 postresection, postradiation, and recurrence. Comparison of = the high=20 versus low tumor burden samples from patients with GBM = demonstrated a trend that was complicated by a single = outlier. This=20 outlier likely reflects the difficulty in truly determining a = "low=20 tumor burden" status based solely on clinical = estimations.=20

The role of YKL-40 in the biology of GBMs is unclear. Given = the=20 relative specificity of increased expression in GBMs relative = to=20 anaplastic gliomas, it is possible that YKL-40 protein is an = integral=20 component of GBM biology and/or may contribute to = GBM-specific=20 histological characteristics, which distinguish them from = grade II=20 and III gliomas. Alternatively, or potentially = complementarily,=20 YKL-40 expression may be the cause or effect of the signaling = abnormalities that characterize GBMs such as elevated Akt or = Ras=20 pathway activity.

The fact that not all of the GBMs express YKL-40 also indicates=20 that it is involved in the biology of only a subset of these=20 tumors, potentially a molecular subdivision of GBMs. It is = possible=20 that for those tumors that do express large amounts of = YKL-40,=20 its levels could be a useful indicator of GBM subtype or = disease=20 status. Although YKL-40 levels showed a wide range of = variability,=20 statistical analysis of this data strongly suggests that in=20 YKL-40 producing high-grade gliomas, serum levels may be a = valuable=20 indicator of disease activity. The utility of the YKL-40 = levels=20 as a surrogate marker of tumor burden, response to treatment, = or relapse remains to be determined. This resolution will = require=20 a prospective trial in which serum samples are analyzed in = patients=20 with tumors demonstrating elevated expression of the = YKL-40=20 gene at the time of diagnosis with subsequent measures during = the course of the illness. Such a prospective study is = planned,=20 evaluating both gene expression in the tumor tissue along = with=20 serial measurement of YKL-40 protein concentrations in serum. =


3D"   =20 ACKNOWLEDGMENTS
 
We = thank Laura K.=20 Schaefer for work on tumor sample preparation for the = Microarray=20 experiment and Western blot analysis. We also thank Michelle = Hammond=20 for help in collecting the serum samples, and Dr. Raymond = Sawaya and=20 the M.D. Anderson Department of Neurosurgery for their = support.=20


3D"   =20 FOOTNOTES
 
The costs of publication of this article were = defrayed=20 in part by the payment of page charges. This article must=20 therefore be hereby marked advertisement in accordance = with 18=20 U.S.C. Section 1734 solely to indicate this fact.

1 Supported by The Bullock=20 Foundation, NIH Grant UO1CA894314-1, the Searle Scholars, = Bressler=20 Scholars, and Tow Foundations.

2 To whom requests for = reprints=20 should be addressed, at Department of Cell Biology, Memorial=20 Sloan-Kettering Cancer Center, 1275 York Avenue, RRL 917B, = New York,=20 NY 10021. Phone: (212) 639-3017; Fax: (646) 422-2062; E-mail: = hollande{at}mskcc.org

3 The abbreviations used = are: GBM,=20 glioblastoma multiforme; ECM, extracellular matrix; TBS-T,=20 Tris-buffered saline with 1% Tween 20.

Received 2/ 4/02. Accepted 5/23/02.


3D"   =20 REFERENCES
Top
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
3D"REFERENCES
&nb= sp;

  1. Kleihues P. Cavenee W. eds. . Pathology and Genetics: = Tumours of=20 the Nervous System, IARC Press Lyon, France 2000.=20
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Opera = requires=0A= the Sun Java Runtime Environment .=0A= =0A= by Andrew Gregory=0A= http://www.scss.com.au/family/andrew/webdesign/xmlhttprequest/=0A= =0A= This work is licensed under the Creative Commons Attribution License. To = view a=0A= copy of this license, visit = http://creativecommons.org/licenses/by-sa/2.5/ or=0A= send a letter to Creative Commons, 559 Nathan Abbott Way, Stanford, = California=0A= 94305, USA.=0A= =0A= Attribution: Leave my name and web address in this script intact.=0A= =0A= Not Supported in Opera=0A= ----------------------=0A= * user/password authentication=0A= * responseXML data member=0A= =0A= Not Fully Supported in Opera=0A= ----------------------------=0A= * async requests=0A= * abort()=0A= * getAllResponseHeaders(), getAllResponseHeader(header)=0A= =0A= */=0A= // IE support=0A= if (window.ActiveXObject && !window.XMLHttpRequest) {=0A= window.XMLHttpRequest =3D function() {=0A= var msxmls =3D new Array(=0A= 'Msxml2.XMLHTTP.5.0',=0A= 'Msxml2.XMLHTTP.4.0',=0A= 'Msxml2.XMLHTTP.3.0',=0A= 'Msxml2.XMLHTTP',=0A= 'Microsoft.XMLHTTP');=0A= for (var i =3D 0; i < msxmls.length; i++) {=0A= try {=0A= return new ActiveXObject(msxmls[i]);=0A= } catch (e) {=0A= }=0A= }=0A= return null;=0A= };=0A= }=0A= // Gecko support=0A= /* ;-) */=0A= // Opera support=0A= if (window.opera && !window.XMLHttpRequest) {=0A= window.XMLHttpRequest =3D function() {=0A= this.readyState =3D 0; // = 0=3Duninitialized,1=3Dloading,2=3Dloaded,3=3Dinteractive,4=3Dcomplete=0A= this.status =3D 0; // HTTP status codes=0A= this.statusText =3D '';=0A= this._headers =3D [];=0A= this._aborted =3D false;=0A= this._async =3D true;=0A= this._defaultCharset =3D 'ISO-8859-1';=0A= this._getCharset =3D function() {=0A= var charset =3D _defaultCharset;=0A= var contentType =3D = this.getResponseHeader('Content-type').toUpperCase();=0A= val =3D contentType.indexOf('CHARSET=3D');=0A= if (val !=3D -1) {=0A= charset =3D contentType.substring(val);=0A= }=0A= val =3D charset.indexOf(';');=0A= if (val !=3D -1) {=0A= charset =3D charset.substring(0, val);=0A= }=0A= val =3D charset.indexOf(',');=0A= if (val !=3D -1) {=0A= charset =3D charset.substring(0, val);=0A= }=0A= return charset;=0A= };=0A= this.abort =3D function() {=0A= this._aborted =3D true;=0A= };=0A= this.getAllResponseHeaders =3D function() {=0A= return this.getAllResponseHeader('*');=0A= };=0A= this.getAllResponseHeader =3D function(header) {=0A= var ret =3D '';=0A= for (var i =3D 0; i < this._headers.length; i++) {=0A= if (header =3D=3D '*' || this._headers[i].h =3D=3D header) {=0A= ret +=3D this._headers[i].h + ': ' + this._headers[i].v + '\n';=0A= }=0A= }=0A= return ret;=0A= };=0A= this.getResponseHeader =3D function(header) {=0A= var ret =3D getAllResponseHeader(header);=0A= var i =3D ret.indexOf('\n');=0A= if (i !=3D -1) {=0A= ret =3D ret.substring(0, i);=0A= }=0A= return ret;=0A= };=0A= this.setRequestHeader =3D function(header, value) {=0A= this._headers[this._headers.length] =3D {h:header, v:value};=0A= };=0A= this.open =3D function(method, url, async, user, password) {=0A= this.method =3D method;=0A= this.url =3D url;=0A= this._async =3D true;=0A= this._aborted =3D false;=0A= this._headers =3D [];=0A= if (arguments.length >=3D 3) {=0A= this._async =3D async;=0A= }=0A= if (arguments.length > 3) {=0A= opera.postError('XMLHttpRequest.open() - user/password not = supported');=0A= }=0A= this.readyState =3D 1;=0A= if (this.onreadystatechange) {=0A= this.onreadystatechange();=0A= }=0A= };=0A= this.send =3D function(data) {=0A= if (!navigator.javaEnabled()) {=0A= alert("XMLHttpRequest.send() - Java must be installed and = enabled.");=0A= return;=0A= }=0A= if (this._async) {=0A= setTimeout(this._sendasync, 0, this, data);=0A= // this is not really asynchronous and won't execute until the = current=0A= // execution context ends=0A= } else {=0A= this._sendsync(data);=0A= }=0A= }=0A= this._sendasync =3D function(req, data) {=0A= if (!req._aborted) {=0A= req._sendsync(data);=0A= }=0A= };=0A= this._sendsync =3D function(data) {=0A= this.readyState =3D 2;=0A= if (this.onreadystatechange) {=0A= this.onreadystatechange();=0A= }=0A= // open connection=0A= var url =3D new java.net.URL(new = java.net.URL(window.location.href), this.url);=0A= var conn =3D url.openConnection();=0A= for (var i =3D 0; i < this._headers.length; i++) {=0A= conn.setRequestProperty(this._headers[i].h, this._headers[i].v);=0A= }=0A= this._headers =3D [];=0A= if (this.method =3D=3D 'POST') {=0A= // POST data=0A= conn.setDoOutput(true);=0A= var wr =3D new = java.io.OutputStreamWriter(conn.getOutputStream(), this._getCharset());=0A= wr.write(data);=0A= wr.flush();=0A= wr.close();=0A= }=0A= // read response headers=0A= // NOTE: the getHeaderField() methods always return nulls for me :(=0A= var gotContentEncoding =3D false;=0A= var gotContentLength =3D false;=0A= var gotContentType =3D false;=0A= var gotDate =3D false;=0A= var gotExpiration =3D false;=0A= var gotLastModified =3D false;=0A= for (var i =3D 0; ; i++) {=0A= var hdrName =3D conn.getHeaderFieldKey(i);=0A= var hdrValue =3D conn.getHeaderField(i);=0A= if (hdrName =3D=3D null && hdrValue =3D=3D null) {=0A= break;=0A= }=0A= if (hdrName !=3D null) {=0A= this._headers[this._headers.length] =3D {h:hdrName, = v:hdrValue};=0A= switch (hdrName.toLowerCase()) {=0A= case 'content-encoding': gotContentEncoding =3D true; break;=0A= case 'content-length' : gotContentLength =3D true; break;=0A= case 'content-type' : gotContentType =3D true; break;=0A= case 'date' : gotDate =3D true; break;=0A= case 'expires' : gotExpiration =3D true; break;=0A= case 'last-modified' : gotLastModified =3D true; break;=0A= }=0A= }=0A= }=0A= // try to fill in any missing header information=0A= var val;=0A= val =3D conn.getContentEncoding();=0A= if (val !=3D null && !gotContentEncoding) = this._headers[this._headers.length] =3D {h:'Content-encoding', v:val};=0A= val =3D conn.getContentLength();=0A= if (val !=3D -1 && !gotContentLength) = this._headers[this._headers.length] =3D {h:'Content-length', v:val};=0A= val =3D conn.getContentType();=0A= if (val !=3D null && !gotContentType) = this._headers[this._headers.length] =3D {h:'Content-type', v:val};=0A= val =3D conn.getDate();=0A= if (val !=3D 0 && !gotDate) this._headers[this._headers.length] = =3D {h:'Date', v:(new Date(val)).toUTCString()};=0A= val =3D conn.getExpiration();=0A= if (val !=3D 0 && !gotExpiration) = this._headers[this._headers.length] =3D {h:'Expires', v:(new = Date(val)).toUTCString()};=0A= val =3D conn.getLastModified();=0A= if (val !=3D 0 && !gotLastModified) = this._headers[this._headers.length] =3D {h:'Last-modified', v:(new = Date(val)).toUTCString()};=0A= // read response data=0A= var reqdata =3D '';=0A= var stream =3D conn.getInputStream();=0A= if (stream) {=0A= var reader =3D new java.io.BufferedReader(new = java.io.InputStreamReader(stream, this._getCharset()));=0A= var line;=0A= while ((line =3D reader.readLine()) !=3D null) {=0A= if (this.readyState =3D=3D 2) {=0A= this.readyState =3D 3;=0A= if (this.onreadystatechange) {=0A= this.onreadystatechange();=0A= }=0A= }=0A= reqdata +=3D line + '\n';=0A= }=0A= reader.close();=0A= this.status =3D 200;=0A= this.statusText =3D 'OK';=0A= this.responseText =3D reqdata;=0A= this.readyState =3D 4;=0A= if (this.onreadystatechange) {=0A= this.onreadystatechange();=0A= }=0A= if (this.onload) {=0A= this.onload();=0A= }=0A= } else {=0A= // error=0A= this.status =3D 404;=0A= this.statusText =3D 'Not Found';=0A= this.responseText =3D '';=0A= this.readyState =3D 4;=0A= if (this.onreadystatechange) {=0A= this.onreadystatechange();=0A= }=0A= if (this.onerror) {=0A= this.onerror();=0A= }=0A= }=0A= };=0A= };=0A= }=0A= // ActiveXObject emulation=0A= if (!window.ActiveXObject && window.XMLHttpRequest) {=0A= window.ActiveXObject =3D function(type) {=0A= switch (type.toLowerCase()) {=0A= case 'microsoft.xmlhttp':=0A= case 'msxml2.xmlhttp':=0A= case 'msxml2.xmlhttp.3.0':=0A= case 'msxml2.xmlhttp.4.0':=0A= case 'msxml2.xmlhttp.5.0':=0A= return new XMLHttpRequest();=0A= }=0A= return null;=0A= };=0A= }=0A= ------=_NextPart_000_003B_01C86B05.8F4FA870 Content-Type: application/octet-stream Content-Transfer-Encoding: quoted-printable Content-Location: http://cancerres.aacrjournals.org/javascript/ajax/utility.js /************************************************************************= *****=0A= * javascript/ajax/utility.js=0A= *=0A= * Utility functions for working with XMLHttpRequest data.=0A= *=0A= * Copyright 2006 Board of Trustees of the Leland Stanford Junior = University.=0A= = *************************************************************************= ***/=0A= =0A= /*=0A= * Copy XML nodes into an HTMLElement. This effectively=0A= * clones XML markup which uses XHTML naming conventions=0A= * into an HTML DOM.=0A= */=0A= function copy_xml_to_html(src, dst) {=0A= if (src.nodeType =3D=3D 1) { /* Node.ELEMENT_NODE */=0A= var e =3D document.createElement(src.nodeName);=0A= for (var i =3D 0; i < src.childNodes.length; i++) {=0A= copy_xml_to_html(src.childNodes[i], e);=0A= }=0A= for (var i =3D 0; i < src.attributes.length; i++) {=0A= var n =3D src.attributes[i].name;=0A= var v =3D unescape_xml_string(src.attributes[i].value); =0A= e.setAttribute(n, v);=0A= if (n =3D=3D "class") {=0A= e.className =3D v;=0A= }=0A= else if (n =3D=3D "style") {=0A= set_css_style(v, e, "");=0A= }=0A= }=0A= dst.appendChild(e);=0A= }=0A= else if (src.nodeType =3D=3D 3) { /* Node.TEXT_NODE */=0A= dst.appendChild(document.createTextNode(src.nodeValue));=0A= }=0A= }=0A= =0A= /* =0A= * It is unclear that this is the right thing to be calling=0A= * from copy_xml_to_html, but it appears that Safari decides=0A= * to convert & to the NCR #, and then encodes that=0A= * NCR to &%26%2338;. So, I'm going to treat the DOM Attr=0A= * value as a plain string, and run our XML string input=0A= * through the decoding routine below.=0A= */=0A= function unescape_xml_string(s) {=0A= return s.replace(/'/g, "'")=0A= .replace(/'/g, "'")=0A= .replace(/"/g, "\"")=0A= .replace(/"/g, "\"")=0A= .replace(/>/g, ">")=0A= .replace(/>/g, ">")=0A= .replace(/</g, "<")=0A= .replace(/</g, "<")=0A= .replace(/&/g, "&")=0A= .replace(/&/g, "&");=0A= }=0A= =0A= /*=0A= * Parse set of CSS rules and apply them to an element.=0A= * This is quite horrifying, but I'm unable to determine=0A= * how else to handle this with IE 6. FireFox and other=0A= * sane browsers let you simply set the style attribute=0A= * or use e.style.setProperty(rule, value, priority),=0A= * IE 6 appears to have neither of these capabilities..=0A= */=0A= function set_css_style(css, e, priority) {=0A= var rules =3D css.split(";");=0A= for (var i =3D 0; i < rules.length; i++) {=0A= var nvpair =3D rules[i].split(":");=0A= if (nvpair.length =3D=3D 2) {=0A= try {=0A= var name =3D nvpair[0]; /* style attribute */=0A= var value =3D nvpair[1]; /* attribute value */=0A= =0A= /*=0A= * For each possible style attribute, set the=0A= * appropriate style property in the element.=0A= */=0A= if (name =3D=3D "background") {=0A= e.style.background =3D value;=0A= }=0A= else if (name =3D=3D "background-attachment") {=0A= e.style.backgroundAttachment =3D value;=0A= }=0A= else if (name =3D=3D "background-color") {=0A= e.style.backgroundColor =3D value;=0A= }=0A= else if (name =3D=3D "background-image") {=0A= e.style.backgroundImage =3D value;=0A= }=0A= else if (name =3D=3D "background-position") {=0A= e.style.backgroundPosition =3D value;=0A= }=0A= else if (name =3D=3D "background-position-x") {=0A= e.style.backgroundPositionX =3D value;=0A= }=0A= else if (name =3D=3D "background-position-y") {=0A= e.style.backgroundPositionY =3D value;=0A= }=0A= else if (name =3D=3D "background-repeat") {=0A= e.style.backgroundRepeat =3D value;=0A= }=0A= else if (name =3D=3D "behavior") {=0A= e.style.behavior =3D value;=0A= }=0A= else if (name =3D=3D "border") {=0A= e.style.border =3D value;=0A= }=0A= else if (name =3D=3D "border-bottom") {=0A= e.style.borderBottom =3D value;=0A= }=0A= else if (name =3D=3D "border-bottom-color") {=0A= e.style.borderBottomColor =3D value;=0A= }=0A= else if (name =3D=3D "border-bottom-style") {=0A= e.style.borderBottomStyle =3D value;=0A= }=0A= else if (name =3D=3D "border-bottom-width") {=0A= e.style.borderBottomWidth =3D value;=0A= }=0A= else if (name =3D=3D "border-collapse") {=0A= e.style.borderCollapse =3D value;=0A= }=0A= else if (name =3D=3D "border-color") {=0A= e.style.borderColor =3D value;=0A= }=0A= else if (name =3D=3D "border-left") {=0A= e.style.borderLeft =3D value;=0A= }=0A= else if (name =3D=3D "border-left-color") {=0A= e.style.borderLeftColor =3D value;=0A= }=0A= else if (name =3D=3D "border-left-style") {=0A= e.style.borderLeftStyle =3D value;=0A= }=0A= else if (name =3D=3D "border-left-width") {=0A= e.style.borderLeftWidth =3D value;=0A= }=0A= else if (name =3D=3D "border-right") {=0A= e.style.borderRight =3D value;=0A= }=0A= else if (name =3D=3D "border-right-color") {=0A= e.style.borderRightColor =3D value;=0A= }=0A= else if (name =3D=3D "border-right-style") {=0A= e.style.borderRightStyle =3D value;=0A= }=0A= else if (name =3D=3D "border-right-width") {=0A= e.style.borderRightWidth =3D value;=0A= }=0A= else if (name =3D=3D "border-style") {=0A= e.style.borderStyle =3D value;=0A= }=0A= else if (name =3D=3D "border-top") {=0A= e.style.borderTop =3D value;=0A= }=0A= else if (name =3D=3D "border-top-color") {=0A= e.style.borderTopColor =3D value;=0A= }=0A= else if (name =3D=3D "border-top-style") {=0A= e.style.borderTopStyle =3D value;=0A= }=0A= else if (name =3D=3D "border-top-width") {=0A= e.style.borderTopWidth =3D value;=0A= }=0A= else if (name =3D=3D "border-width") {=0A= e.style.borderWidth =3D value;=0A= }=0A= else if (name =3D=3D "bottom") {=0A= e.style.bottom =3D value;=0A= }=0A= else if (name =3D=3D "clear") {=0A= e.style.clear =3D value;=0A= }=0A= else if (name =3D=3D "clip") {=0A= e.style.clip =3D value;=0A= }=0A= else if (name =3D=3D "color") {=0A= e.style.color =3D value;=0A= }=0A= else if (name =3D=3D "cssText") {=0A= e.style.Sets =3D value;=0A= }=0A= else if (name =3D=3D "cursor") {=0A= e.style.cursor =3D value;=0A= }=0A= else if (name =3D=3D "direction") {=0A= e.style.direction =3D value;=0A= }=0A= else if (name =3D=3D "display") {=0A= e.style.display =3D value;=0A= }=0A= else if (name =3D=3D "font") {=0A= e.style.font =3D value;=0A= }=0A= else if (name =3D=3D "font-family") {=0A= e.style.fontFamily =3D value;=0A= }=0A= else if (name =3D=3D "font-size") {=0A= e.style.fontSize =3D value;=0A= }=0A= else if (name =3D=3D "font-style") {=0A= e.style.fontStyle =3D value;=0A= }=0A= else if (name =3D=3D "font-variant") {=0A= e.style.fontVariant =3D value;=0A= }=0A= else if (name =3D=3D "font-weight") {=0A= e.style.fontWeight =3D value;=0A= }=0A= else if (name =3D=3D "height") {=0A= e.style.height =3D value;=0A= }=0A= else if (name =3D=3D "ime-mode") {=0A= e.style.imeMode =3D value;=0A= }=0A= else if (name =3D=3D "layout-flow") {=0A= e.style.layoutFlow =3D value;=0A= }=0A= else if (name =3D=3D "layout-grid") {=0A= e.style.layoutGrid =3D value;=0A= }=0A= else if (name =3D=3D "layout-grid-char") {=0A= e.style.layoutGridChar =3D value;=0A= }=0A= else if (name =3D=3D "layout-grid-line") {=0A= e.style.layoutGridLine =3D value;=0A= }=0A= else if (name =3D=3D "layout-grid-mode") {=0A= e.style.layoutGridMode =3D value;=0A= }=0A= else if (name =3D=3D "layout-grid-type") {=0A= e.style.layoutGridType =3D value;=0A= }=0A= else if (name =3D=3D "left") {=0A= e.style.left =3D value;=0A= }=0A= else if (name =3D=3D "letter-spacing") {=0A= e.style.letterSpacing =3D value;=0A= }=0A= else if (name =3D=3D "line-break") {=0A= e.style.lineBreak =3D value;=0A= }=0A= else if (name =3D=3D "line-height") {=0A= e.style.lineHeight =3D value;=0A= }=0A= else if (name =3D=3D "list-style") {=0A= e.style.listStyle =3D value;=0A= }=0A= else if (name =3D=3D "list-style-image") {=0A= e.style.listStyleImage =3D value;=0A= }=0A= else if (name =3D=3D "list-style-position") {=0A= e.style.listStylePosition =3D value;=0A= }=0A= else if (name =3D=3D "list-style-type") {=0A= e.style.listStyleType =3D value;=0A= }=0A= else if (name =3D=3D "margin") {=0A= e.style.margin =3D value;=0A= }=0A= else if (name =3D=3D "margin-bottom") {=0A= e.style.marginBottom =3D value;=0A= }=0A= else if (name =3D=3D "margin-left") {=0A= e.style.marginLeft =3D value;=0A= }=0A= else if (name =3D=3D "margin-right") {=0A= e.style.marginRight =3D value;=0A= }=0A= else if (name =3D=3D "margin-top") {=0A= e.style.marginTop =3D value;=0A= }=0A= else if (name =3D=3D "min-height") {=0A= e.style.minHeight =3D value;=0A= }=0A= else if (name =3D=3D "overflow") {=0A= e.style.overflow =3D value;=0A= }=0A= else if (name =3D=3D "overflow-x") {=0A= e.style.overflowX =3D value;=0A= }=0A= else if (name =3D=3D "overflow-y") {=0A= e.style.overflowY =3D value;=0A= }=0A= else if (name =3D=3D "padding") {=0A= e.style.padding =3D value;=0A= }=0A= else if (name =3D=3D "padding-bottom") {=0A= e.style.paddingBottom =3D value;=0A= }=0A= else if (name =3D=3D "padding-left") {=0A= e.style.paddingLeft =3D value;=0A= }=0A= else if (name =3D=3D "padding-right") {=0A= e.style.paddingRight =3D value;=0A= }=0A= else if (name =3D=3D "padding-top") {=0A= e.style.paddingTop =3D value;=0A= }=0A= else if (name =3D=3D "page-break-after") {=0A= e.style.pageBreakAfter =3D value;=0A= }=0A= else if (name =3D=3D "page-break-before") {=0A= e.style.pageBreakBefore =3D value;=0A= }=0A= else if (name =3D=3D "pixelBottom") {=0A= e.style.pixelBottom =3D value;=0A= }=0A= else if (name =3D=3D "pixelHeight") {=0A= e.style.pixelHeight =3D value;=0A= }=0A= else if (name =3D=3D "pixelLeft") {=0A= e.style.pixelLeft =3D value;=0A= }=0A= else if (name =3D=3D "pixelRight") {=0A= e.style.pixelRight =3D value;=0A= }=0A= else if (name =3D=3D "pixelTop") {=0A= e.style.pixelTop =3D value;=0A= }=0A= else if (name =3D=3D "pixelWidth") {=0A= e.style.pixelWidth =3D value;=0A= }=0A= else if (name =3D=3D "posBottom") {=0A= e.style.posBottom =3D value;=0A= }=0A= else if (name =3D=3D "posHeight") {=0A= e.style.posHeight =3D value;=0A= }=0A= else if (name =3D=3D "position") {=0A= e.style.position =3D value;=0A= }=0A= else if (name =3D=3D "posLeft") {=0A= e.style.posLeft =3D value;=0A= }=0A= else if (name =3D=3D "posRight") {=0A= e.style.posRight =3D value;=0A= }=0A= else if (name =3D=3D "posTop") {=0A= e.style.posTop =3D value;=0A= }=0A= else if (name =3D=3D "posWidth") {=0A= e.style.posWidth =3D value;=0A= }=0A= else if (name =3D=3D "right") {=0A= e.style.right =3D value;=0A= }=0A= else if (name =3D=3D "ruby-align") {=0A= e.style.rubyAlign =3D value;=0A= }=0A= else if (name =3D=3D "ruby-overhang") {=0A= e.style.rubyOverhang =3D value;=0A= }=0A= else if (name =3D=3D "ruby-position") {=0A= e.style.rubyPosition =3D value;=0A= }=0A= else if (name =3D=3D "scrollbar-3dlight-color") {=0A= e.style.scrollbar3dLightColor =3D value;=0A= }=0A= else if (name =3D=3D "scrollbar-arrow-color") {=0A= e.style.scrollbarArrowColor =3D value;=0A= }=0A= else if (name =3D=3D "scrollbar-base-color") {=0A= e.style.scrollbarBaseColor =3D value;=0A= }=0A= else if (name =3D=3D "scrollbar-darkshadow-color") {=0A= e.style.scrollbarDarkShadowColor =3D value;=0A= }=0A= else if (name =3D=3D "scrollbar-face-color") {=0A= e.style.scrollbarFaceColor =3D value;=0A= }=0A= else if (name =3D=3D "scrollbar-highlight-color") {=0A= e.style.scrollbarHighlightColor =3D value;=0A= }=0A= else if (name =3D=3D "scrollbar-shadow-color") {=0A= e.style.scrollbarShadowColor =3D value;=0A= }=0A= else if (name =3D=3D "scrollbar-track-color") {=0A= e.style.scrollbarTrackColor =3D value;=0A= }=0A= else if (name =3D=3D "float") {=0A= e.style.styleFloat =3D value;=0A= }=0A= else if (name =3D=3D "table-layout") {=0A= e.style.tableLayout =3D value;=0A= }=0A= else if (name =3D=3D "text-align") {=0A= e.style.textAlign =3D value;=0A= }=0A= else if (name =3D=3D "text-align-last") {=0A= e.style.textAlignLast =3D value;=0A= }=0A= else if (name =3D=3D "text-autospace") {=0A= e.style.textAutospace =3D value;=0A= }=0A= else if (name =3D=3D "text-decoration") {=0A= e.style.textDecoration =3D value;=0A= }=0A= else if (name =3D=3D "textDecorationBlink") {=0A= e.style.textDecorationBlink =3D value;=0A= }=0A= else if (name =3D=3D "textDecorationLineThrough") {=0A= e.style.textDecorationLineThrough =3D value;=0A= }=0A= else if (name =3D=3D 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"vertical-align") {=0A= e.style.verticalAlign =3D value;=0A= }=0A= else if (name =3D=3D "visibility") {=0A= e.style.visibility =3D value;=0A= }=0A= else if (name =3D=3D "white-space") {=0A= e.style.whiteSpace =3D value;=0A= }=0A= else if (name =3D=3D "width") {=0A= e.style.width =3D value;=0A= }=0A= else if (name =3D=3D "word-break") {=0A= e.style.wordBreak =3D value;=0A= }=0A= else if (name =3D=3D "word-spacing") {=0A= e.style.wordSpacing =3D value;=0A= }=0A= else if (name =3D=3D "word-wrap") {=0A= e.style.wordWrap =3D value;=0A= }=0A= else if (name =3D=3D "writing-mode") {=0A= e.style.writingMode =3D value;=0A= }=0A= else if (name =3D=3D "z-index") {=0A= e.style.zIndex =3D value;=0A= }=0A= else if (name =3D=3D "zoom") {=0A= e.style.zoom =3D value;=0A= }=0A= }=0A= catch (e) {=0A= /* ignore error on attempt to set e.style.[property] */=0A= }=0A= }=0A= }=0A= }=0A= ------=_NextPart_000_003B_01C86B05.8F4FA870 Content-Type: application/octet-stream Content-Transfer-Encoding: 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