bluuebunny/crossref_metadata_2025 huggingface dataset:
Integrin‐linked kinase expression increases with ovarian tumour grade and is sustained by peritoneal tumour fluid
Authors: Ahmed Nuzhat , Riley Clyde , Oliva Karen , Stutt Emma , E Rice Greg , A Quinn Michael
Published: September 2003

Integrin‐linked kinase (ILK) is a serine threonine kinase, overexpression of which promotes tumour growth and invasion through deregulation of the cell cycle. This study demonstrates the relative expression of ILK in normal, benign, low‐grade, and high‐grade (borderline, grade I/II, and grade III) ovarian tumours of serous, mucinous, endometrioid, and clear cell types in order to assess its potential as a marker for epithelial ovarian cancer progression. Seventy‐three specimens including ten normal, ten benign, 14 borderline, 17 grade I/II, and 22 grade III were evaluated by immunohistochemistry. Immunoreactive ILK was not detectable in normal ovarian surface epithelium. All 53 carcinomas studied were positive and the staining intensity correlated significantly with the grade of the tumour. Ovarian cancer cell lines had high expression of ILK, while immortalized normal ovarian surface epithelial cell lines (HOSE) showed low basal expression of ILK by western blotting. Peritoneal tumour fluid (PTF) upregulated ILK expression in ovarian cancer cell lines but had no effect on HOSE cells. PTF‐induced up‐regulation of ILK expression in ovarian cancer cell lines correlated with the activation of the downstream protein kinase B (PKB/Akt) pathway. Collectively, these data demonstrate that ILK expression increases with ovarian cancer progression and that soluble factors in PTF mediate sustained overexpression of ILK in ovarian cancer cells. Suppression of ILK expression may therefore represent a novel and an efficient mechanism for controlling ovarian tumour growth. Copyright © 2003 John Wiley & Sons, Ltd.
Epstein–Barr virus infection in colorectal neoplasms associated with inflammatory bowel disease: detection of the virus in lymphomas but not in adenocarcinomas
Authors: Wong NACS , Herbst H , Herrmann K , Kirchner T , Krajewski AS , Moorghen M , Niedobitek F , Rooney N , Shepherd NA , Niedobitek G
Published: September 2003

Epstein–Barr virus (EBV) is associated with several lymphoid and epithelial human malignancies. The latter include gastric adenocarcinomas, while sporadic colorectal adenocarcinomas (CRCs) have been reported to be EBV‐negative. Recently, increased numbers of EBV‐infected B lymphocytes have been detected in intestinal mucosal samples affected by ulcerative colitis (UC) and, to a lesser extent, Crohn's disease (CD). Both CRC and colorectal non‐Hodgkin's lymphoma (NHL) are recognized complications of inflammatory bowel disease (IBD), but it is unclear to what extent EBV contributes to the development of these neoplasms. Seventeen cases of IBD‐associated CRC and nine cases of IBD‐associated colorectal NHL were therefore studied for the presence of EBV by in situ hybridization. EBV‐positive cases were further studied for the expression of the EBV‐encoded nuclear antigen (EBNA) 2 and the latent membrane protein (LMP) 1 of EBV by immunohistochemistry. Four out of seven cases of colorectal NHL associated with UC were shown to be EBV‐positive. In addition, two of two colorectal NHLs developing in patients with CD were EBV‐positive. Of the EBV‐positive lymphomas, three displayed a pattern of EBV latent gene expression consistent with type I latency (EBNA2−/LMP1−), two a type II pattern (EBNA2−/LMP1+), and one a type III pattern (EBNA2+/LMP1+). These findings suggest that EBV infection is involved in the pathogenesis of a proportion of colorectal NHLs developing in IBD. Iatrogenic immunosuppression may contribute to the development of these lymphomas. By contrast, all 17 IBD‐associated CRCs were EBV‐negative, including a case of CRC occurring synchronously with an EBV‐positive NHL. In conjunction with previous reports on sporadic CRCs, this suggests that EBV is not involved in the pathogenesis of CRC. Copyright © 2003 John Wiley & Sons, Ltd.
Differential expression of CD146 in tissues and endothelial cells derived from infantile haemangioma and normal human skin
Authors: Li Qing , Yu Ying , Bischoff Joyce , Mulliken John B , Olsen Bjorn R
Published: September 2003

Haemangioma is the most common tumour of endothelial origin, occurring in 4–10% of Caucasian infants. It is characterized by rapid growth during the first year of postnatal life, followed by spontaneous regression from 1 to 7 years of age. The cell surface adhesion molecule CD146 has been identified as an endothelial cell marker. Despite advances in understanding the functional role of CD146 in normal endothelial cells and tumour progression, its expression and a possible role in an endothelial tumour have not been studied. As part of an investigation of endothelial cell alterations in infantile haemangioma, differential expression studies were performed with several known antigens and endothelial cell markers. Using immunohistochemical and flow cytometric analyses, cultured human dermal microvascular endothelial cells isolated from newborn foreskin (HDMEC) were compared with endothelial cells derived from haemangioma tissue (HemECs). In addition, immunohistochemistry was used to compare haemangioma tissues with normal human skin. Unexpectedly, cultured HemECs showed a significantly lower level of CD146 than HDMECs by both flow cytometric analysis and immunofluorescence staining. Using immunohistochemical studies, it was further demonstrated that endothelia in all haemangioma tissues, regardless of the tumour phase, showed negative immunoreactivity for CD146. In contrast, strong positive staining for CD146 was observed in the pericyte‐like cells that surround the endothelial layers. These findings are believed to be relevant to the molecular basis of haemangioma. Furthermore, it is possible that antibodies against CD146 may be useful for separating haemangioma‐derived endothelial cells from normal endothelial cells and pericytes. Copyright © 2003 John Wiley & Sons, Ltd.
A novel immunohistochemical method to estimate cell‐cycle phase distribution in archival tissue: implications for the prediction of outcome in colorectal cancer
Authors: Scott Ian S , Morris Lesley S , Bird Kate , Davies R Justin , Vowler Sarah L , Rushbrook Simon M , Marshall Aileen E , Laskey Ronald A , Miller Richard , Arends Mark J , Coleman Nicholas
Published: September 2003

An immunohistochemical method for assessing cell‐cycle phase distribution in colorectal resection specimens would enable phase data to be incorporated into diagnostic algorithms for the estimation of prognosis and response to adjuvant chemotherapy in colorectal cancer. In contrast to flow cytometry, an immunohistochemical method would also allow the phase distribution to be examined within morphologically heterogeneous regions of neoplasms. Paraffin sections of normal colon (n = 25), colonic adenoma (n = 15), and colonic adenocarcinoma (n = 30) were analysed by immunohistochemistry using antibodies against markers of cell‐cycle entry, Mcm‐2 and Ki67, and putative markers of the cell‐cycle phase, cyclins D1 and E (putative markers of G1 phase), cyclin A (S phase), cytoplasmic cyclin B1 (G2 phase), and phosphohistone H3 (M phase). The phase specificity of each marker was assessed by examining the degree of co‐expression of adjacent phase markers using double‐antibody fluorescence confocal microscopy and by comparison with flow cytometric analysis performed on adjacent tissue sections. The S‐phase specificity of detectable cyclin A was also assessed in combination with in situ DNA replication using fluorescence confocal microscopy. All cells expressing phase markers co‐expressed Mcm‐2. Adjacent phase markers were not significantly co‐expressed, confirming the relative specificity of these markers in tissue sections of colon. Cell‐cycle phase distribution, calculated by immunohistochemistry, compared well with phase analyses obtained by flow cytometry. No cells expressed cyclin A in the absence of active DNA replication. The S‐phase labelling index, as defined by detectable cyclin A expression, showed a positive correlation with the Mcm‐2 labelling index and increased in the progression from normal colon to adenocarcinoma. In conclusion, a combination of these cell‐cycle phase markers can be used to calculate the distribution of cells throughout each phase of the cell cycle in colorectal tissue sections. Detectable cyclin A can be used as a surrogate marker of S phase and may be of value in predicting prognosis and response to adjuvant therapy. Copyright © 2003 John Wiley & Sons, Ltd.
Detection of apoptosis using in situ markers for DNA strand breaks in the failing human heart. Fact or epiphenomenon?
Authors: Hughes Siân E
Published: September 2003

The role of apoptosis and the methods used for its detection in failing human hearts are controversial. Recent data have challenged the hypothesis that in situ markers for DNA strand breaks mirror apoptotic (TUNEL and Taq in situ ligation assay) and/or necrotic (Pfu in situ ligation assay) cell death, and thus provide evidence that apoptotic cell loss contributes to the progression of heart failure. Experimental data cast doubt not only upon the specificity of the TUNEL technique but also the Taq in situ ligation assay as a reliable method for the detection of apoptotic cell death and provide compelling new evidence that the occurrence of cardiomyocyte cell death as defined by the detection of DNA strand breaks using either TUNEL or Taq and Pfu in situ ligation assays is an epiphenomena that is not related to the evolution of heart failure. Cardiomyocyte positivity for in situ markers of DNA strand breaks is a feature of hypertrophic cardiomyopathic hearts, irrespective of the underlying pathology or the presence or absence of heart failure. These data raise concerns regarding the extent of apoptosis in cardiomyopathy and the contribution of this process to the progression of heart failure. Copyright © 2003 John Wiley & Sons, Ltd.
Beta‐catenin accumulation in the progression of human hepatocarcinogenesis correlates with loss of E‐cadherin and accumulation of p53, but not with expression of conventional WNT‐1 target genes
Authors: Prange Wilhelm , Breuhahn Kai , Fischer Frank , Zilkens Christoph , Pietsch Torsten , Petmecky Katharina , Eilers Renate , Dienes Hans‐Peter , Schirmacher Peter
Published: September 2003

Beta‐catenin integrates intracellular WNT signalling and the intercellular E‐cadherin–catenin adhesion system. To date, little is known about the role of β‐catenin activation and nuclear accumulation in hepatocarcinogenesis. This study has analysed β‐catenin expression patterns in human dysplastic nodules (DNs), as well as in hepatocellular carcinomas (HCCs) in comparison with proliferation, expression of WNT‐1 target genes, E‐cadherin, and p53. One hundred and seventy HCCs and 25 DNs were categorized according to established criteria and analysed for the expression pattern of β‐catenin. Analysis of the proliferative activity and expression of E‐cadherin, cyclin D1, MMP‐7, c‐myc, and p53 was performed on a representative subgroup of cases. All DNs lacked nuclear β‐catenin, while 36% of all HCCs were positive, with the number of nuclear stained cells ranging from less than 1% to more than 90%. Increasing nuclear accumulation of β‐catenin correlated with reduced membranous E‐cadherin expression and nuclear p53 but not with proliferation. Cyclin D1, MMP‐7, and c‐myc expression was detected in 54%, 26%, and 65% of HCCs, respectively, but did not correlate with nuclear β‐catenin, proliferation, or grading. Sequence analysis of the β‐catenin gene revealed no detectable mutations in DNs, but mutations in the GSK‐3β binding site were present in 14.3% of the HCCs. In conclusion, this study has demonstrated that nuclear accumulation of β‐catenin is a frequent progression event in human hepatocarcinogenesis which correlates with nuclear p53 accumulation and loss of membranous E‐cadherin, but not with the expression pattern of established WNT‐1 target genes. It is hypothesized that the role of β‐catenin in human HCC differs significantly from its established function in colon carcinogenesis. Copyright © 2003 John Wiley & Sons, Ltd.
WIF1, a component of the Wnt pathway, is down‐regulated in prostate, breast, lung, and bladder cancer
Authors: Wissmann Christoph , Wild Peter Johannes , Kaiser Simone , Roepcke Stefan , Stoehr Robert , Woenckhaus Matthias , Kristiansen Glen , Hsieh Jen‐Chih , Hofstaedter Ferdinand , Hartmann Arndt , Knuechel Ruth , Rosenthal André , Pilarsky Christian
Published: September 2003

To detect novel Wnt‐pathway genes involved in tumourigenesis, this study analysed the RNA expression levels of 40 genes of the Wnt pathway by chip hybridization of microdissected matched pairs of 54 primary prostate carcinomas. Eleven genes showed greater than two‐fold differential expression in at least 10% of prostate cancers. Three of these genes encode extracellular components of the Wnt pathway (WNT2, WIF1, SFRP4); two are receptors (FZD4, FZD6); two belong to the intracellular signal cascade (DVL1, PPP2CB); one regulates transcription (TCF4); and three represent genes regulated by this pathway (CCND2, CD44, MYC). While SFRP4, FZD4, FZD6, DVL1, TCF4, and MYC are up‐regulated, WIF1, WNT2, PPP2CB, CCND2, and CD44 are down‐regulated in certain prostate cancer patients. Wnt inhibitory factor 1 (WIF1) and secreted frizzled related protein (SFRP4) showed the most significant aberrant expression at the RNA level. WIF1 was down‐regulated in 64% of primary prostate cancers, while SFRP4 was up‐regulated in 81% of the patients. Immunohistochemical analysis using a polyclonal antibody revealed strong cytoplasmic perinuclear WIF1 expression in normal epithelial cells of the prostate, breast, lung, and urinary bladder. Strong reduction of WIF1 protein expression was found in 23% of prostate carcinomas, but also in 60% of breast, 75% of non‐small cell lung (NSCLC), and 26% of bladder cancers analysed. No significant association between WIF1 down‐regulation and tumour stage or grade was observed for prostate, breast or non‐small cell lung carcinomas, indicating that loss of WIF1 expression may be an early event in tumourigenesis in these tissues. However, down‐regulation of WIF1 correlated with higher tumour stage in urinary bladder tumours (pTa versus pT1–pT4; p = 0.038). Copyright © 2003 John Wiley & Sons, Ltd.
FANCD2 protein is expressed in proliferating cells of human tissues that are cancer‐prone in Fanconi anaemia
Authors: Hölzel Michael , van Diest Paul J , Bier Patrick , Wallisch Michael , Hoatlin Maureen E , Joenje Hans , de Winter Johan P
Published: September 2003

Fanconi anaemia (FA) is an inherited form of progressive pancytopenia associated with developmental defects, chromosomal instability, and cancer predisposition. At least seven distinct FA proteins function in concert to protect the genome, a key step being the activation of FANCD2 by mono‐ubiquitination. This paper reports an immunohistochemical analysis of FANCD2 expression in normal human tissue. The highest expression was observed in maturing spermatocytes and fetal oocytes (consistent with a role for FANCD2 in meiosis) and in germinal centre cells of the spleen, tonsil, and lymph nodes (consistent with a role in proliferation). FANCD2 expression was also seen in tissues predisposed to cancer development in FA patients: haematopoietic cells, especially in the fetus, and squamous cell epithelia, particularly in the head and neck region and uterine cervix. FANCD2 expression was also occasionally seen in the breast and Fallopian tube epithelium, the respiratory epithelium of the trachea, and the exocrine cells of the pancreas, indicating that these tissues may also be cancer‐prone in FA. FANCD2 expression is frequently expressed in proliferating cells as demonstrated by Ki‐67 immunofluorescence double staining, consistent with a function of FANCD2 in DNA replication. Copyright © 2003 John Wiley & Sons, Ltd.
Quantitative RT‐PCR in cirrhotic nodules reveals gene expression changes associated with liver carcinogenesis
Authors: Colombat Magali , Paradis Valérie , Bièche Ivan , Dargère Delphine , Laurendeau Ingrid , Belghiti Jacques , Vidaud Michel , Degott Claude , Bedossa Pierre
Published: September 2003

Cirrhosis is considered to be the precursor of most hepatocellular carcinomas. To gain insight into the early molecular mechanisms of liver carcinogenesis, this study compared, using real‐time quantitative reverse transcriptase‐polymerase chain reaction (RT‐PCR), the expression levels of 31 selected genes in normal livers, cirrhotic nodules, and hepatocellular carcinomas. Since cirrhosis is composed of a mixture of polyclonal and monoclonal nodules, gene expression levels were also compared according to the clonal status of the cirrhotic nodules. The expression of eight of the 31 genes studied was significantly increased (NEGF2, ANGPT1, ARF, KRT19, SFN, CLDN4, MMP7, and ETV4) in cirrhotic nodules compared with normal liver, while only one was decreased (LYVE1). The same trend of variation was observed in cirrhosis and hepatocellular carcinomas for all of these genes except KRT19. When gene expression variation was compared according to the clonal status of cirrhotic nodules, only the LYVE1 expression level was significantly different. The LYVE1 gene expression level decreased progressively from polyclonal cirrhotic nodules to monoclonal cirrhotic nodules (polyclonal nodules 0.39 ± 0.25; monoclonal nodules 0.20 ± 0.14; p < 0.05) and to hepatocellular carcinoma (0.07 ± 0.1). In conclusion, this study highlights the fact that among genes strongly dysregulated in hepatocellular carcinoma, some are already abnormally expressed in cirrhosis. The decrease in the expression level of one of these genes, LYVE1, was associated with monoclonality in cirrhotic nodules. Copyright © 2003 John Wiley & Sons, Ltd.
Compositional heterogeneity effects in multisticker associative polyelectrolytes prepared by micellar polymerization
Authors: Kujawa Piotr , Audibert‐Hayet Annie , Selb Joseph , Candau Françoise
Published: September 2003

An aqueous micellar radical polymerization technique has been used to prepare water‐soluble polyelectrolytes of acrylamide (AM) and sodium 2‐acrylamido‐2‐methylpropanesulfonate (NaAMPS) hydrophobically modified with low amounts (2–4 mol %) of N,N‐dihexylacrylamide (DiHexAM). This synthesis method leads to multisticker polymers, in which the number and length of the hydrophobic blocks vary with the degree of conversion. Kinetic studies show an appreciable compositional heterogeneity for these polymers, in contrast to their neutral analogues (i.e., AM/DiHexAM copolymers). The drift in composition observed for the AM/NaAMPS/DiHexAM terpolymers is attributed to the electrostatic repulsion between the negatively charged growing polymeric chains, located in the aqueous continuous medium, and the anionic sodium dodecyl sulfate micelles, inside of which the hydrophobic monomer is entrapped. It is shown that an optimization of the polymerization process can be achieved, which gives terpolymers of homogeneous and well‐defined compositions. The thickening ability of the hydrophobically modified polyelectrolytes is directly affected by the extent of their compositional heterogeneity: an increase in the sample homogeneity leads to a significant viscosity enhancement and marked elastic properties of the polymer solutions. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 3261–3274, 2003
Complex polymer architectures through dimethoxymethyl 1,1‐diphenylethylene derivatives
Authors: Meyer Nicolas , Eddine Mohamad Nasser , Delaite Christelle , Hurtrez Guy , Dumas Philippe
Published: September 2003

The reactions of polystyryllithium and potassium on dimethoxymethyl 1,1‐diphenylethylene derivatives were studied in different solvents. In a polar medium, A3 and A6 star types were formed according to the stoichiometry, whereas in a nonpolar medium, hyperbranched structures were synthesized. Extensions of an already proposed mechanism in polar and nonpolar media were examined. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 3329–3335, 2003
Synthesis and characterization of poly(methyl methacrylate)/casein nanoparticles with a well‐defined core‐shell structure
Authors: Zhu Junmin , Li Pei
Published: September 2003

Well‐defined, core‐shell poly(methyl methacrylate) (PMMA)/casein nanoparticles, ranging from 80 to 130 nm in diameter, were prepared via a direct graft copolymerization of methyl methacrylate (MMA) from casein. The polymerization was induced by a small amount of alkyl hydroperoxide (ROOH) in water at 80 °C. Free radicals on the amino groups of casein and alkoxy radicals were generated concurrently, which initiated the graft copolymerization and homopolymerization of MMA, respectively. The presence of casein micelles promoted the emulsion polymerization of the monomer and provided particle stability. The conversion and grafting efficiency of the monomer strongly depended on the type of radical initiator, ROOH concentration, casein to MMA ratio, and reaction temperature. The graft copolymers and homopolymer of PMMA were isolated and characterized with Fourier transform infrared spectroscopy and differential scanning calorimetry. The molecular weight determination of both the grafted and homopolymer of PMMA suggested that the graft copolymerization and homopolymerization of MMA proceeded at a similar rate. The transmission electron microscopic image of the nanoparticles clearly showed a well‐defined core‐shell morphology, where PMMA cores were coated with casein shells. The casein shells were further confirmed with a zeta‐potential measurement. Finally, this synthetic method allowed us to prepare PMMA/casein nanoparticles with a solid content of up to 31%. Thus, our new process is commercially viable. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 3346–3353, 2003
Synthesis and characterization of novel polyimides with 2,2‐bis[4(4‐aminophenoxy)phenyl]phthalein‐3′,5′‐bis(trifluoromethyl)anilide
Authors: Myung B. Y. , Kim J. S. , Kim J. J. , Yoon T. H.
Published: September 2003

2,2‐Bis[4(4‐aminophenoxy)phenyl]phthalein‐3′,5′‐bis(trifluoromethyl)anilide (6FADAP), containing fluorine and phthalimide moieties, was synthesized via the Williamson ether condensation reaction from 1‐chloro‐4‐nitrobenzene and phenolphthalein‐3′,5′‐bis(trifluoromethyl)anilide, which was followed by hydrogenation. Monomers such as 2,2‐bis[4(4‐aminophenoxy)phenyl]phthalein‐anilide containing phthalimide groups and 2,2‐bis[4(4‐aminophenoxy)phenyl]phthalein containing only phthalein moieties were also synthesized for comparison. The monomers were first characterized by Fourier transform infrared (FTIR), 1H NMR, 19F NMR, elemental analysis, and titration and were then used to prepare polyimides with 2,2‐bis(3,4‐dicarboxyphenyl)hexafluoropropane dianhydride. The polyimides were designed to have molecular weights of 20,000 g/mol via off‐stoichiometry and were characterized by FTIR, NMR, gel permeation chromatography (GPC), differential scanning calorimetry, and thermogravimetric analysis. Their solubility, water absorption, dielectric constant, and refractive index were also evaluated. The polyimides prepared with 6FADAP, containing fluorine and phthalimide moieties, had excellent solubility in N‐methylpyrrolidinone, N,N‐dimethylacetamide, tetrahydrofuran, CHCl3, tetrachloroethane, and acetone, and GPC analysis showed a molecular weight of 18,700 g/mol. The polyimides also exhibited a high glass‐transition temperature (290 °C), good thermal stability (∼500 °C in air), low water absorption (1.9 wt %), a low dielectric constant (2.81), a low refractive index, and low birefringence (0.0041). © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 3361–3374, 2003
Ethylene polymerization with porous polystyrene spheres supported Cp2ZrCl2 catalyst
Authors: Qin Yongxin , Tang Tao , Zhao Zhongfu , Huang Baotong
Published: September 2003

A catalyst with porous polystyrene beads supported Cp2ZrCl2 was prepared and tested for ethylene polymerization with methylaluminoxane as a cocatalyst. By comparison, the porous supported catalyst maintained higher activity and produced polyethylene with better morphology than its corresponding solid supported catalyst. The differences between activities of the catalysts and morphologies of the products were reasonably explained by the fragmentation processes of support as frequently observed with the inorganic supported Ziegler–Natta catalysts. Investigation into the distribution of polystyrene in the polyethylene revealed the fact that the porous polystyrene supported catalyst had undergone fragmentation during polymerization. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 3313–3319, 2003
Synthesis and chiral recognition ability of optically active poly{N‐[(R)‐α‐methoxycarbonylbenzyl]methacrylamide} with various tacticities by radical polymerization using Lewis acids
Authors: Morioka Kohei , Suito Yoshikatsu , Isobe Yutaka , Habaue Shigeki , Okamoto Yoshio
Published: September 2003

The radical polymerization of an optically active methacrylamide, N‐[(R)‐α‐methoxycarbonylbenzyl]methacrylamide, was carried out in the absence and presence of Lewis acids such as yittribium trifluoromethanesulfonate [Yb(OTf)3] and scandium trifluoromethanesulfonate [Sc(OTf)3]. Catalytic amounts of the Lewis acids significantly affected the stereoregularity of the obtained polymers. The polymerization with Yb(OTf)3 in tetrahydrofuran afforded isotactic polymers (up to mm = 87%), whereas the conventional radical method without the Lewis acid produced polymers rich in syndiotacticity (up to rr = 88%). The radical polymerization in the presence of MgBr2 proceeded in a heterotactic‐selective manner (mr = 63%). Thus, the isotactic, syndiotactic, and heterotactic poly(methacrylamide)s were synthesized by the radical processes. The chiral recognition abilities of the obtained optically active poly(methacrylamide)s were affected by the stereoregularity. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 3354–3360, 2003
Synthesis of well‐defined macromonomers and comb copolymers from polymers made by atom transfer radical polymerization
Authors: Muehlebach Andreas , Rime François
Published: September 2003

Poly(n‐butyl acrylate) macromonomers with predetermined molecular weights (1300 < number‐average molecular weight < 23,000) and low polydispersity indices (<1.2) were synthesized from bromine‐terminated atom transfer radical polymerization polymers via end‐group substitution with acrylic acid and methacrylic acid. These macromonomers, having a high degree of end‐group functionalization (>90%), were radically homopolymerized to obtain comb polymers. A high macromonomer concentration, combined with a low radical flux, was needed to obtain a high conversion of the macromonomers and a reasonable degree of polymerization. By the traditional radical copolymerization of the hydrophobic macromonomers with the hydrophilic monomer N,N‐dimethylaminoethyl methacrylate (DMAEMA), amphiphilic comb copolymers were obtained. The conversions of the macromonomers and comonomer were almost quantitative under optimized reaction conditions. The molecular weights were high (number‐average molecular weight ≈70,000), and the molecular weight distribution was broad (polydispersity index ≈ 3.5). Kinetic measurements showed simultaneous decreases in the macromonomer and DMAEMA concentrations, indicating a relatively homogeneous composition of the comb copolymers over the whole molecular weight range. This was supported by preparative size exclusion chromatography. The copolymerization of poly(n‐butyl acrylate) macromonomers with other hydrophilic monomers such as acrylic acid or N,N‐dimethylacrylamide gave comb copolymers with multimodal molecular weight distributions in size exclusion chromatography and extremely high apparent molecular weights. Dynamic light scattering showed a heterogeneous composition consisting of small (6–9 nm) and large (23–143 nm) particles, probably micelles or other type of aggregates. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 3425–3439, 2003
Cationic photopolymerization of epoxynorbornane linseed oils: The effect of diluents
Authors: Zong Zhengang , Soucek Mark D. , Liu Yubiao , Hu Jun
Published: September 2003

New epoxynorbornane linseed oils (ENLOs) were prepared as a function of the norbornene content. The cationic photopolymerization of ENLOs was investigated with real‐time infrared spectroscopy and photo‐differential scanning calorimetry. The effects of reactive and nonreactive diluents on the polymerization rate were also studied. The diluents reduced the viscosity of the formulations, markedly accelerated the rate of polymerization of ENLOs, and increased their final conversions. The effects of the reactive diluent were compared for ENLOs and epoxidized linseed oil. The relative reactivity of the oxiranes was not as important as the viscosity of the reacting system, and it was proposed that the cationic photopolymerization of ENLOs was controlled by diffusion. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 3440–3456, 2003
Pneumonia due to Chlamydia pneumoniae in children: Epidemiology, diagnosis, and treatment
Authors: Hammerschlag Margaret R.
Published: September 2003

The term “atypical” pneumonia has been used to differentiate infections caused by Chlamydia pneumoniae, Mycoplasma pneumoniae, Legionella, and other related organisms from pneumonia caused by classic bacteria, the prototype being Streptococcus pneumoniae. However, recent studies demonstrated that the clinical presentation of pneumonia due to atypical pathogens cannot readily be differentiated from those caused by “typical” bacteria. This is further complicated by the observation that coinfections with atypical pathogens and other bacteria are frequent. Nonetheless, the term “atypical” can be useful, as these organisms share a number of characteristics that separate them from “typical” bacteria. They are either obligate or facultative intracellular parasites that cannot be isolated using routine microbiologic methods. The most commonly used method of diagnosis of these infections is serology, which has significant limitations. Although C. pneumoniae is now recognized worldwide as a common cause of respiratory infections in adults and children, major gaps remain in our knowledge of the biology of this organism and how it causes disease, in major part due to the lack of readily available, standardized diagnostic methods. Pediatr Pulmonol. 2003; 36:384–390. © 2003 Wiley‐Liss, Inc.
Pediatric lymphocytic interstitial pneumonitis in an HIV‐negative child with pulmonary Epstein‐Barr virus infection
Authors: Mueller Gary A. , Pickoff Arthur S.
Published: September 2003

Lymphocytic interstitial pneumonitis (LIP) in children has been most commonly associated with human immunodeficiency virus (HIV) infection. Epstein‐Barr virus (EBV) associated LIP without HIV infection has been reported only in adults. EBV associated LIP has been reported in children, but only with concurrent HIV infection. We report a case of EBV associated, HIV negative LIP in a child. Pediatr Pulmonol. 2003; 36:447–449. © 2003 Wiley‐Liss, Inc.
Exhaled nitric oxide and tracheal endothelin‐1 in preterm infants with and without RDS
Authors: Olsen Steven L. , Clark Perry L. , Thibeault Donald W. , Norberg Mike , Truog William E.
Published: September 2003

We measured exhaled nitric oxide and tracheal aspirate endothelin‐1 to determine relationships between these substances and alterations in pulmonary gas exchange during respiratory distress syndrome (RDS) in comparison to those obtained from control preterm infants without RDS. Eight infants with RDS had measurements made at 24 hr and again at 48–72 hr. Eight control infants were studied once at 24–48 hr of life. Exhaled gas was analyzed on‐line, and minute excretion of NO (V̇NO) was calculated. ET‐1 was determined by immunoassay. Median V̇NO at 24 hr in RDS was 0.405 nl/min/kg (range, 0.30 –0.79), which subsequently declined by 48–72 hr to 0.166 nl/min/kg (P < 0.01). The V̇NO in RDS infants was significantly higher than time‐matched V̇NO in controls, with a median of 0.099 nl/min/kg (range, 0.03–0.27; P < 0.001). ET‐1 was not correlated with initial V̇NO in the RDS or control patients. In conclusion, in RDS, V̇NO decreases as gas exchange improves. ET‐1 is detectable in tracheal aspirate samples in both groups of infants. Pediatr Pulmonol. 2003; 36:421–426. © 2003 Wiley‐Liss, Inc.
Pseudomonas aeruginosa and cystic fibrosis: Correlation between exoenzyme production and patient's clinical state
Authors: Lanotte P. , Mereghetti L. , Lejeune B. , Massicot P. , Quentin R.
Published: September 2003

In this study, we investigated the correlation between the production by Pseudomonas aeruginosa isolates of four exoenzymes (protease, elastase, neuraminidase, and phospholipase C (PLC)) and the clinical state of cystic fibrosis (CF) patients. We studied 212 P. aeruginosa isolates from 22 CF patients chronically infected with this bacterium. Patients were classified into three clinical groups according to a modified Shwachman‐Kulczycki‐Khaw (SKK) scoring system. The production of enzymes by isolates from patients in the three populations was analyzed and compared using four statistical tests: chi‐square, Mann‐Whitney U, principal component analysis, and discriminant analysis. Isolates from patients with excellent or good clinical status (group I, SKK score ≥71) had higher elastase and neuraminidase activities than isolates from the other patients. In contrast, PLC activity, a common characteristic of CF isolates, was higher in isolates from patients with poor or weak clinical status (group III, SKK score ≤55). PLC also appeared to be the best parameter for differentiating between groups I and III. Enzyme production was highly variable in group II isolates (SKK score, 56–70). Our results suggest that P. aeruginosa isolates from patients with good clinical status produce large amounts of neuraminidase, and that PLC production may be involved in the decrease in pulmonary function. Pediatr Pulmonol. 2003; 36:405–412. © 2003 Wiley‐Liss, Inc.
Effect of long‐acting β2 agonists on exacerbation rates of asthma in children
Authors: Bisgaard Hans
Published: September 2003

The purpose of this analysis was to examine the effect of long‐acting β2‐adrenoceptor agonists (LABAs) on the asthma exacerbation rate in pediatric patients. Randomized controlled trials (RCT) that included the use of LABAs to treat symptoms of pediatric asthma in children on inhaled corticosteroids, that reported asthma exacerbation rates, and that were published as full papers in peer‐reviewed journals were retrieved from a search of the medical literature. Eight studies were identified that fulfilled these criteria. An exacerbation was defined as deterioration in a patient's asthma requiring a change in prescribed medication or not defined but reported as an asthma exacerbation or an asthma‐related hospitalization. Analysis of data from the eight studies revealed no apparent protection from an asthma exacerbation among children on a LABA compared to patients on comparator treatment. The relative risk of an asthma exacerbation for LABA compared to placebo or short‐acting β2‐adrenoceptor agonist (SABA) ranged from 0.95–1.86. The relative risk of hospitalization for asthma in patients treated with LABAs with regular maintenance with ICS ranged from 3.3–21.6 in the three studies that reported asthma‐related hospitalizations. The lack of evidence for the control of asthma exacerbations in children regularly using a LABA should bring into question its general use as add‐on therapy. Studies should be designed to directly explore the implications of these observations in pediatric patients. Pediatr Pulmonol. 2003; 36:391–398. © 2003 Wiley‐Liss, Inc.
Epidemic spread of Pandoraea apista, a new pathogen causing severe lung disease in cystic fibrosis patients
Authors: Jørgensen Inger Merete , Johansen Helle Krogh , Frederiksen Birgitte , Pressler Tacjana , Hansen Annelise , Vandamme Peter , Høiby Niels , Koch Christian
Published: September 2003

We show that Pandoraea apista must be added to the increasing list of pathogens capable of causing chronic lung infection in cystic fibrosis (CF) patients. It is most likely that this strain of P. apista was transmissible among patients with CF, leading to spread of infection from the index patient to 5 other patients exposed during participation in winter camps and/or hospitalization. All patients developed chronic infection with high levels of antibodies, and 4 patients had a downhill course of lung disease. P. apista must therefore be considered a new and sometimes important pathogen for CF patients. Cohort isolation prevented further spread of P. apista in our CF center. Pediatr Pulmonol. 2003; 36:439–446. © 2003 Wiley‐Liss, Inc
Molecular characterization of human prostate carcinoma cell lines
Authors: van Bokhoven Adrie , Varella‐Garcia Marileila , Korch Christopher , Johannes Widya U. , Smith E. Erin , Miller Heidi L. , Nordeen Steven K. , Miller Gary J. , Lucia M. Scott
Published: September 2003

BACKGROUND

This study presents a comprehensive survey and characterization of available prostate carcinoma cell lines, most of which have been widely used but are incompletely characterized.

METHODS

A total of 21 cell lines were investigated, including three “classical” (DU 145, LNCaP, and PC‐3) and 18 “non‐classical” lines (1013L, 22Rv1, ALVA‐55, ALVA‐101, ARCaP, CWR‐R1, DuCaP, DuPro‐1, LAPC‐4, MDA PCa 1, MDA PCa 2a, MDA PCa 2b, NCI‐H660, PC‐346C, PC‐93, PSK‐1, UM‐SCP‐1, and VCaP). Cytogenetics, DNA profiling, expression of basal, luminal, and neuroendocrine differentiation markers, and mutation analyses of the TP53 and androgen receptor (AR) genes were performed.

RESULTS

Based on cytogenetics and DNA profiling analyses, out of the 18 “non‐classical” lines, six were confirmed to be unique, eight (in four pairs) were confirmed to be related in origin, and four lines were identified as cross‐contaminants. Of this latter group, PC‐93 was found to be a derivative of HeLa, whereas DuPro‐1, ALVA‐55, and ALVA‐101 were derivatives of PC‐3. The 17 genuine prostate cell lines expressed keratin 8 (K8) and K18. Nine showed AR expression, of which five harbored mutations in the AR gene. Prostate‐specific antigen and DD3 were exclusively detected in AR expressing cell lines. Seven lines expressed the basal cell marker K5, three of these lines showed co‐expression of AR.

CONCLUSIONS

This study defines a collection of 17 genuine prostate carcinoma cell lines. This collection, although small, constitutes a variety of different types and stages of prostate cancer, while it also partly reflects the heterogeneous nature of this malignancy. Prostate 57: 205–225, 2003. © 2003 Wiley‐Liss, Inc.
Spectral karyotype (SKY) analysis of human prostate carcinoma cell lines
Authors: van Bokhoven Adrie , Caires Aimee , Maria Michael Di , Schulte Aline Passarini , Lucia M. Scott , Nordeen Steven K. , Miller Gary J. , Varella‐Garcia Marileila
Published: September 2003

BACKGROUND

Well‐characterized in vitro model systems provide an invaluable tool for studying prostate cancer in the laboratory. Detailed karyotypes have been reported using modern techniques such as multiplex fluorescence in situ hybridization (M‐FISH) and spectral karyotyping (SKY) for LNCaP, DU 145, NCI‐H660, and PC‐3 cell lines. However, karyotypic data for more recently established prostate carcinoma cell lines are still limited.

METHODS

Classical (G‐banding) and SKY analyses were performed on ten prostate carcinoma cell lines: 22Rv1, CWR‐R1, DuCaP, LAPC‐4, MDA PCa 1, MDA PCa 2a, MDA PCa 2b, PC‐346C, PSK‐1, and VCaP.

RESULTS

Chromosomal abnormalities were identified in all cell lines, although the number and complexity varied greatly among them. PC‐346C, established from a primary tumor, exhibited the lowest number (3) of clonal structural abnormalities, while DuCaP, established from a metastasis from a hormone‐refractory patient, exhibited both the highest number (31) and largest complexity of structural abnormalities. In various subsets of these models, breakpoints were identified in chromosomal regions previously described as being involved in prostate cancer (e.g., 8p, 10q, 13q, and 16q).

CONCLUSIONS

The present study provides a comprehensive karyotypic analysis of a large number of prostate carcinoma cell lines, and offers a valuable resource for future investigations. Prostate 57: 226–244, 2003. © 2003 Wiley‐Liss, Inc.
Transactivation of epidermal growth factor receptor after heparin‐binding epidermal growth factor‐like growth factor shedding in the migration of prostate cancer cells promoted by bombesin
Authors: Madarame Jun , Higashiyama Shigeki , Kiyota Hiroshi , Madachi Ayako , Toki Fujio , Shimomura Tatsuya , Tani Naoyuki , Oishi Yukihiko , Matsuura Nariaki
Published: September 2003

BACKGROUND

A pathway consisting of bombesin, G‐protein coupling receptors (GPCRs), metalloproteases, pro‐heparin‐binding epidermal growth factor (proHB‐EGF), and epidermal growth factor receptor (EGFR) has been reported in prostate cancer cells. The occurrence of HB‐EGF shedding from proHB‐EGF in this pathway, however, has not been proven directly. In addition, it is still unclear how much this pathway contributes to the migration of prostate cancer cells. In this study, we tried to directly elucidate HB‐EGF shedding in this pathway and to determine its contribution to the migration of prostate cancer cells.

METHODS

RT‐PCR and indirect immunofluorescence staining for HB‐EGF and its receptors, such as EGFR and HER4/erbB4, were performed on PC‐3 cells. The influences of bombesin, anti‐EGFR neutralizing monoclonal antibody, HB‐EGF, and HB‐EGF shedding inhibitor on the migration of PC‐3 cells were studied by means of in vitro wound assays. The amount of HB‐EGF shed from PC‐3 cells with alkaline phosphatase‐tagged HB‐EGF in the presence of bombesin was determined by measuring AP activity. Immunoprecipitations and phosphotyrosine Western blotting were performed to detect EGFR transactivated by bombesin.

RESULTS

PC‐3 expressed HB‐EGF and EGFR, but not HER4/erbB4. PC‐3 migrated in the presence of bombesin, but its migration was partly inhibited by the neutralizing antibody against EGFR. PC‐3 also migrated in the presence of HB‐EGF, but HB‐EGF shedding inhibitor partly inhibited this phenomenon. HB‐EGF was shed from PC‐3 cells in the presence of bombesin, and this shedding was inhibited by HB‐EGF shedding inhibitor. In addition, the EGFR on PC‐3 was activated in the presence of bombesin and inactivated in the presence of HB‐EGF shedding inhibitor.

CONCLUSIONS

These results indicated that HB‐EGF shedding and the following transactivation of EGFR occurs in this pathway and that this pathway partly contributes to the migration of prostate cancer cells. Prostate 57: 187–195, 2003. © 2003 Wiley‐Liss, Inc.
Expression of protein C inhibitor (PCI) in benign and malignant prostatic tissues
Authors: Cao Yue , Becker Charlotte , Lundwall Åke , Christensson Anders , Gadaleanu Virgil , Lilja Hans , Bjartell Anders
Published: September 2003

BACKGROUND

Protein C inhibitor (PCI) occurs at high concentration in seminal plasma, and inhibits human glandular kallikrein‐2 and, less readily, prostate‐specific antigen. Previous studies have localized PCI in the male genital tract. Here we have performed a detailed investigation of PCI expression in the prostatic tissues, metastases, and cell lines.

METHODS

Immunohistochemistry, in situ hybridization, and Western blotting were used to study prostatic tissues, metastases, and PC‐3, DU‐145, and LNCaP cells.

RESULTS

PCI was immunolocalized in tissue microarray spots with BPH epithelium (detection rate 100%), PIN lesions (100%), tumors (96%), metastases (88%), and in all cell lines. ISH and WB supported the findings.

CONCLUSIONS

PCI is widely expressed in benign prostatic epithelium, and may act as a local regulator of enzymatic activity in seminal fluid, of importance for normal sperm function. Lack of PCI expression in a subpopulation of high‐grade tumor cells in combination with maintained protease expression may facilitate invasive growth patterns. Prostate 57: 196–204, 2003. © 2003 Wiley‐Liss, Inc.
Increased fatty acid synthase expression and activity during progression of prostate cancer in the TRAMP model
Authors: Pflug Beth R. , Pecher Stefana M. , Brink Alisa W. , Nelson Joel B. , Foster Barbara A.
Published: September 2003

BACKGROUND

Fatty acid synthase (FAS) is the major enzyme required to convert carbohydrates to fatty acids. Recent evidence suggests that FAS activity is essential for prostate cancer growth and survival, since blocking the enzyme activity results in cell death. In this study, the role of FAS up‐regulation during prostate tumor progression in the transgenic adenocarcinoma of mouse prostate (TRAMP) model was investigated. Sensitivity to FAS anti‐metabolites was also analyzed in TRAMP prostate tumor cells and tissue to determine therapeutic potential of FAS inhibition in the treatment of prostate cancer.

METHODS

FAS expression was evaluated by immunohistochemistry of TRAMP tissues, including primary and metastatic lesions in mice of varying ages. FAS pathway activity was studied in vitro using TRAMP‐derived cell lines and in vivo in TRAMP tissues. The sensitivity of TRAMP cell lines and tissues to the antimetabolite drugs (2R,3S)‐2,3‐epoxy‐4‐oxo‐7,10‐trans, transdodecadienamide (cerulenin) and C‐75, which target FAS, was determined by FAS antimetabolite inhibition of 14C‐acetate conversion to fatty acids, cell growth inhibition, and apoptosis analyses.

RESULTS

High FAS expression and activity in the TRAMP mouse prostate was evident at 12 weeks of age compared with nontransgenic littermates and further increased with age, tumor progression, and in metastatic lesions. FAS pathway inhibition resulted in a dose‐dependent reduction in cell survival and decreased enzyme activity in these models.

CONCLUSIONS

These data suggest that the up‐regulation of FAS expression play a role in tumorigenesis of the prostate in the TRAMP model and hence can provide valuable insight into human prostate cancer. Given the response of tumor cells to FAS antimetabolites, FAS may serve as a novel target for prostate cancer therapy. Prostate 57: 245–254, 2003. © 2003 Wiley‐Liss, Inc.