bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of c-ErbB1/Epidermal Growth Factor Receptor and c-ErbB2/HER-2 Expression in Bronchial Dysplasia: Evaluation of Potential Targets for Chemoprevention of Lung Cancer
Authors: Merrick Daniel T. , Kittelson John , Winterhalder Ralph , Kotantoulas Georgia , Ingeberg Steen , Keith Robert L. , Kennedy Timothy C. , Miller York E. , Franklin Wilbur A. , Hirsch Fred R.
Published: April 2006

Purpose: Lung cancer is preceded by a premalignant phase during which intervention could decrease associated morbidity and mortality. Molecular characterization of factors involved in controlling progression of bronchial dysplasias will provide markers of premalignant change and identify targets for chemoprevention.

Experimental Design: Immunohistochemical analysis of epidermal growth factor receptor (EGFR; c-ErbB1/EGFR), HER-2/neu (c-ErbB2/HER-2), Ki-67, and minichromosome maintenance protein 2 (MCM2) expression in bronchial dysplasia was undertaken to characterize molecular alterations associated with the progression of these lesions in 268 bronchoscopically obtained biopsies from 134 subjects.

Results: Analysis of biopsies with the most severe diagnosis from each subject showed a linear relationship between increasing marker expression and severity of dysplastic change for EGFR (P < 0.001), Ki-67 (P < 0.001), and MCM2 (P = 0.001) but not HER-2 (P = 0.102). Increased expression of either EGFR or HER-2 was associated with increased levels of Ki-67 and MCM2 expression, and combined overexpression of these receptors was associated with the highest levels of proliferation, suggesting a synergistic effect. Finally, the lack of an associated trend toward increased EGFR expression when comparing the worst and best biopsies within each subject indicated a potential field effect in the induction of EGFR expression.

Conclusions: The results suggest a prominent role for EGFR overexpression in the development and progression of bronchial dysplasia and provide rationale for exploring inhibition of EGFR signaling in lung cancer chemoprevention.
Antisense Suppression of Pygopus2 Results in Growth Arrest of Epithelial Ovarian Cancer
Authors: Popadiuk Cathy M. , Xiong Jieying , Wells Malcolm G. , Andrews Phillip G. , Dankwa Kweku , Hirasawa Kensuke , Lake Blue B. , Kao Kenneth R.
Published: April 2006

Purpose: The Pygopus proteins are critical elements of the canonical Wnt/β-catenin transcriptional complex. In epithelial ovarian cancer, constitutively active Wnt signaling is restricted to one (endometrioid) tumor subtype. The purpose of this study was to determine the level of expression and growth requirements of human Pygopus2 (hPygo2) protein in epithelial ovarian cancer.

Experimental Design: Expression and subcellular localization of hPygo2 was determined in epithelial ovarian cancer cell lines and tumors using Northern blot, immunoblot, and immunofluorescence. Immunohistochemistry was done on 125 archived patient epithelial ovarian cancer tumors representing all epithelial ovarian cancer subtypes. T-cell factor–dependent transcription levels were determined in epithelial ovarian cancer cells using TOPflash/FOPflash in vivo assays. Phosphorothioated antisense oligonucleotides were transfected into cell lines and growth assayed by cell counting, anchorage-independent colony formation on soft agar, and xenografting into severe combined immunodeficient mice.

Results: All six epithelial ovarian cancer cell lines and 82% of the patient samples overexpressed nuclear hPygo2 compared with control cells and benign disease. Depletion of hPygo2 by antisense oligonucleotides in both Wnt-active (TOV-112D) and Wnt-inactive serous (OVCAR-3, SKOV-3) and clear cell (TOV-21G) carcinoma cell lines halted growth, assessed using tissue culture, anchorage-independent, and xenograft assays.

Conclusions: hPygo2 is unexpectedly widely expressed in, and required in the absence of, Wnt signaling for malignant growth of epithelial ovarian cancer, the deadliest gynecologic malignancy. These findings strongly suggest that inhibition of hPygo2 may be of therapeutic benefit for treating this disease.
Combination Analysis of Hypermethylated Wnt-Antagonist Family Genes as a Novel Epigenetic Biomarker Panel for Bladder Cancer Detection
Authors: Urakami Shinji , Shiina Hiroaki , Enokida Hideki , Kawakami Toshifumi , Kawamoto Ken , Hirata Hiroshi , Tanaka Yuichiro , Kikuno Nobuyuki , Nakagawa Masayuki , Igawa Mikio , Dahiya Rajvir
Published: April 2006

Purpose: Aberrant promoter hypermethylation of Wnt-antagonist genes contributes to the pathogenesis of several cancers. We hypothesized that combined methylation analysis of Wnt-antagonist genes could improve their use as a panel of biomarkers for diagnosing and staging of bladder cancers.

Experimental Design: Samples (54 total) of bladder tumor and corresponding normal bladder mucosa were analyzed for the methylation and expression levels of six Wnt-antagonist genes (sFRP-1, sFRP-2, sFRP-4, and sFRP-5, Wif-1, and Dkk-3). To increase the sensitivity/specificity of bladder tumor detection, the methylation score (M score), a new method for multigene methylation analysis, was developed. The M score of each sample was calculated as the sum of the corresponding log hazard ratio coefficients derived from multivariate logistic regression analysis of the methylation status for each Wnt-antagonist gene. Receiver operator characteristic (ROC) curve analysis was used to determine the optimal sensitivity/specificity of the M score. Urine DNA from 24 matched patients with bladder tumor and 20 cancer-free volunteers was also used to investigate the methylation status of Wnt-antagonist genes.

Results: The methylation levels of Wnt-antagonists were significantly higher and mRNA levels were significantly lower in bladder tumor than in bladder mucosa. Each methylation level was inversely correlated with the corresponding mRNA level. In multivariate regression analysis, the methylation levels of sFRP-2 and Dkk-3 were significant independent predictors of bladder tumor (P < 0.05 and P < 0.01, respectively), whereas with sFRP-1, sFRP-5, and Wif-1 there was a trend towards significance as independent predictors. The M score of Wnt-antagonist genes was significantly higher in bladder tumor than in bladder mucosa (P < 0.05). Overall, the M score had a sensitivity of 77.2% and a specificity of 66.7% as a diagnostic biomarker (areas under the curve, 0.763). The M score could distinguish superficial from invasive bladder tumors with a sensitivity of 72.2% and a specificity of 61.1% as a staging biomarker (areas under the curve, 0.671). In patients with bladder tumor, 80.6% of the methylation-specific PCR results had identical methylation in samples of tumor- and urine-derived DNA. Most urine DNA in normal controls showed no aberrant methylation of the Wnt-antagonist genes.

Conclusions: Hypermethylation of Wnt-antagonist genes plays an important role in the pathogenesis of bladder tumor and can be detected using cellular DNA extracted from urine samples. This is the first report demonstrating that M score analysis of Wnt-antagonist genes could serve as an excellent epigenetic biomarker panel for bladder tumors.
Chemotherapy and Targeted Therapy Combinations in Advanced Melanoma
Authors: Flaherty Keith T.
Published: April 2006

For three decades, clinical trials with chemotherapy in melanoma have failed to show superiority of any one regimen over another. Dacarbazine remains the only “standard” agent. With response rates of <10% and median progression-free survival of 2 months or less in contemporary trials, there is a need to improve systemic therapy. Combination chemotherapy is associated with higher response rates than single-agent therapy but this has not translated into improved survival. An increasing number of potential therapeutic targets have been identified. For some, pharmacologic inhibitors are available, including sorafenib for BRAF, farnesyltransferase inhibitors for NRAS, PD-0325901 for mitogen-activated protein kinase/extracellular signal-regulated kinase kinase, rapamycin analogues for mammalian target of rapamycin, and agents that inhibit either vascular endothelial growth factor or its receptors. Several multitargeted kinase inhibitors have potency against the fibroblast growth factor receptor, c-kit, and platelet-derived growth factor receptor. Small-molecule inhibitors of c-met and Akt are in preclinical development. Another class of agents indirectly affect aberrant signaling, including inhibitors of chaperones and proteasomes. Several targeted agents seem to enhance the cytotoxicity of chemotherapy in preclinical models. The mechanism by which signaling inhibition might synergize with chemotherapy requires more study so that rational combinations move forward. Very few targeted agents have been studied rigorously in this fashion.
Sentinel Lymph Node Biopsy and Melanoma Biology
Authors: Essner Richard
Published: April 2006

Minimally invasive intraoperative lymphatic mapping and sentinel node biopsy has become the standard approach for staging the regional lymph nodes for early-stage melanoma. The procedure requires close collaboration of surgeon, pathologist, and nuclear medicine physician. The strength of lymphatic mapping and sentinel node biopsy is its accuracy of detecting occult lymph node metastases. Reverse transcriptase-PCR (RT-PCR) analyses of either fresh-frozen or paraffin-embedded sections of the sentinel lymph nodes have been found to be more sensitive than H&E staining or immunohistochemistry techniques, but lack of specificity and limits in the availability of tissue specimens make this technique impractical for routine use. Three randomized clinical trials are examining the therapeutic value of lymphatic mapping and sentinel node biopsy for melanoma. Preliminary results of the Multicenter Lymphadenectomy Trial I show the high level of accuracy and low morbidity of lymphatic mapping and sentinel node biopsy done through an international working group. The therapeutic value of lymphatic mapping and sentinel node biopsy is still unclear. Multicenter Lymphadenectomy Trial II will test the clinical significance of lymph nodes evaluated by RT-PCR and the value of completion lymph node dissection for patients found to have tumor-positive sentinel lymph nodes by H&E, immunohistochemistry, or RT-PCR. The Sunbelt Melanoma Trial examines the therapeutic value of completion dissection and benefits of Intron A. The ability to detect occult nodal metastases and evaluate the interaction of primary tumor with the regional lymph nodes may provide for better understanding of the metastatic process in patients with melanoma and help to determine the function of the regional lymph nodes as markers of metastases or incubators of tumor cells in the metastatic cascade.
Identifying and Overcoming Immune Resistance Mechanisms in the Melanoma Tumor Microenvironment
Authors: Gajewski Thomas F.
Published: April 2006

The continually growing list of defined tumor antigens is broadening the potential applicability of tumor antigen-targeted cancer therapies. Although cancer vaccines and adoptive T-cell transfer have been shown to increase the frequency of circulating tumor antigen-specific T cells, these approaches cause clinical responses in a few patients. In melanoma, approximately one third of metastatic lesions contain activated T cells, including those specific for tumor antigens, arguing that the priming phase has occurred already in such individuals even without vaccination. These observations indicate that tumor resistance to immune destruction may dominate in many instances, arguing for a thorough analysis of the melanoma tumor microenvironment in individual patients. Recent work has suggested that T-cell anergy, the influence of CD4+CD25+ regulatory T cells, the expression of inhibitory ligands, such as PD-L1, and the activity of nutrient-catabolizing enzymes, such as indoleamine 2,3-dioxygenase, may be involved. Preclinical murine models have shown that interfering with each of these processes can translate into T-cell–mediated tumor control. Importantly, each of these targets is amenable to clinical manipulation. Clinical translation of these approaches to counter negative regulation of antitumor immunity should receive high priority.
Genetic Alterations in Signaling Pathways in Melanoma
Authors: Haluska Frank G. , Tsao Hensin , Wu Helen , Haluska Frank S. , Lazar Alexander , Goel Vikas
Published: April 2006

Alterations in the RAS signaling cascade are almost uniformly present in melanoma. RAS itself is only infrequently mutated in melanoma although downstream of RAS lie BRAF on the mitogen-activated protein kinase pathway and PTEN on the protein kinase B/Akt pathway. These genes are often altered in melanomas; indeed, the most frequent target of mutation in melanomas is BRAF, which is mutated in ∼60% to 70% of superficial spreading melanomas. These mutations occur in a background that is not normal, with the CDKN2A locus also typically being mutated. We review herein the data that suggest that the distribution of the signaling mutations is important. In general, melanomas carry a mutated NRAS, a mutated BRAF, or concurrent BRAF and PTEN mutations. These data support the hypothesis that the biochemical functions of RAS are portioned by mutations in the pathways lying downstream. Moreover, these mutations have no apparent relationship to the patterns of alteration of CDKN2A and its downstream effectors. Thus, the data also suggest that successful exploitation of mutations in melanoma will be dependent on understanding not only mutations and their frequency but their genetic context as well.
Low and High Tenascin-Expressing Tumors Are Efficiently Targeted by ST2146 Monoclonal Antibody
Authors: De Santis Rita , Albertoni Claudio , Petronzelli Fiorella , Campo Silvia , D'Alessio Valeria , Rosi Antonio , Anastasi Anna Maria , Lindstedt Ragnar , Caroni Nadia , Arseni Brunilde , Chiodi Pierino , Verdoliva Antonio , Cassani Giovanni , Chinol Marco , Paganelli Giovanni , Carminati Paolo
Published: April 2006

ST2146biot is a biotinylated anti-tenascin monoclonal antibody (mAb) to be used for Pretargeted Antibody Guided Radioimmunotherapy (PAGRIT) of solid tumors. In vivo biodistribution studies of 125I-labeled ST2146biot were done in nude mice transplanted with human HT-29 colon carcinoma and/or human U-118MG glioblastoma cells characterized for low and high tenascin expression, respectively. In vitro results show that ST2146 retains immunoreactivity upon biotinylation, in contrast to other anti-tenascin mAbs. In vivo biodistribution of ST2146 shows specific tumor accumulation up to 10 days after the i.v. injection, with no relevant differences between biotinylated and nonbiotinylated ST2146. A dose of 4 μg/mouse saturates the low tenascin-expressing human colon carcinoma HT-29, whereas the high tenascin-expressing human glioblastoma U-118MG seems to be saturated at a ST2146biot dose between 320 and 640 μg/mouse. The percentage of injected dose per gram of tumor ranges from 10% to 30%, corresponding to an amount of ST2146biot/g of tumor of ∼400 ng/g and >200 μg/g for HT-29 and U-118MG, respectively. Tumor to normal organs uptake ratios are between 15 and 60, confirming high tumor selectivity of ST2146biot despite its cross-reactivity with the tenascin expressed at low level in the normal mouse organs. The ST2146biot localization data are substantially confirmed even when both low and high tenascin-expressing tumors are implanted in the same animal. To our knowledge, the absolute amount of ST2146biot, specifically localized in xenotransplanted human tumors, is the highest thus far described and supports the clinical use of this mAb in PAGRIT®.
Celecoxib Up-Regulates the Expression of the ζ Chain of T Cell Receptor Complex in Tumor-Infiltrating Lymphocytes in Human Cervical Cancer
Authors: Ferrandina Gabriella , Ranelletti Franco Oreste , Legge Francesco , Salutari Vanda , Martinelli Enrica , Fattorossi Andrea , Lorusso Domenica , Zannoni Gianfranco , Vellone Valerio , Paglia Amelia , Scambia Giovanni
Published: April 2006

Purpose: We evaluated the effects of celecoxib treatment on tumor-infiltrating lymphocyte (TIL) subsets [CD3+, CD4+,CD8+, CD25+, and T cell receptor (TCR)-ζ–expressing cells] and tryptase-positive mast cells in cervical tumors. Circulating levels of cytokines [interleukin (IL)-1β, IL-10, tumor necrosis factor-α, IL-6, and IL-12] and angiogenesis-modulating factors (vascular endothelial growth factor and endostatin) have also been analyzed.

Experimental Design: Cervical tumor biopsies and blood samples were obtained at the time of diagnosis and after 10 days of celecoxib treatment (400 mg b.i.d., at 8:00 a.m. and 8:00 p.m.) in 27 cases. Immunohistochemistry and ELISA assays were used to assess the expression of biological factors in tumor tissue and circulating levels of cytokines and angiogenic molecules.

Results: We showed a statistically significant increase in the percentage of TIL expressing the TCR-ζ chain after celecoxib treatment: indeed, in cases exposed to celecoxib, the percentage of TCR-ζ+ cells ranged from 5.0 to 50.0 (median, 22.5) with respect to baseline expression (range, 3.0-50.0; median, 10.0; P = 0.0016). There was no significant treatment-related difference in the percentage of CD3+, CD4+, CD8+, and CD25+ TIL as well as in tryptase-positive cells. IL-12 levels were significantly reduced in posttreatment samples with respect to baseline levels (P = 0.002). We also found a reduction in the circulating levels of vascular endothelial growth factor, and a statistically significant increase of serum endostatin levels (P = 0.035).

Conclusions: We reported the first evidence in humans that celecoxib restores ζ expression by TIL in primary cervical tumors, suggesting that a positive modulation of immune function may serve as an additional mechanism supporting the antitumor effect of this class of drugs.
Immunosuppression in Melanoma Immunotherapy: Potential Opportunities for Intervention
Authors: Lizée Gregory , Radvanyi Laszlo G. , Overwijk Willem W. , Hwu Patrick
Published: April 2006

Although melanomas are substantially more immunogenic than other tumors, current immunotherapeutic approaches for melanoma patients have met with only limited success. Although melanoma-specific CD8+ T-cell responses can often be generated in patients naturally or through vaccination regimens, tumors frequently continue to grow unabated, suggesting that tumor-specific immune responses may be actively dampened in vivo. Research over the past decade has brought to light several mechanisms used by melanomas and other tumors to suppress tumor-specific immune responses. These include the presence of regulatory immune cells within the tumor microenvironment and draining lymph nodes that serve to shut down effector T-cell function. In addition, melanoma tumors themselves express a number of soluble and membrane-bound molecules that are responsible for inhibiting activated immune cells. The identification of these suppressive mechanisms has provided significant opportunities for designing novel therapeutic interventions that could augment current vaccination and adoptive transfer approaches for treatment of melanoma.
Allogeneic and Autologous Melanoma Vaccines: Where Have We Been and Where Are We Going?
Authors: Sondak Vernon K. , Sabel Michael S. , Mulé James J.
Published: April 2006

The past three decades have seen substantial research on vaccines for the treatment of metastatic melanoma and the prevention of recurrence following resection. Despite their enormous promise, the actual results have been disappointing, with several high-profile vaccine clinical trials failing to show a benefit. Nonetheless, enthusiasm for melanoma vaccines remains and has increased with our expanding understanding of the immune response to tumor. Cellular vaccines can be divided into autologous, derived from the patient's own tumor and allogeneic vaccines. Autologous vaccines have the advantage of containing all potentially relevant tumor-associated antigens for that particular patient. However, autologous vaccines are difficult to obtain from most patients with advanced disease and impossible to obtain from patients who present after resection of all clinically evident disease. No consensus exists for how tumors should be processed, preserved, modified, and delivered to serve as an effective vaccine. The amount of autologous tumor available is rarely enough to produce more than two or three vaccination doses, and the time between initial tumor harvest and ultimate availability of the vaccine may result in interval tumor progression that diminishes the likelihood of vaccine efficacy. All these drawbacks of autologous tumor vaccination limit its applicability and also limit the ability to test autologous vaccines in prospective trials. Allogeneic vaccines avoid many of these problems, but may not contain all of the tumor-associated antigens present on the patient's own tumor. In particular, neoantigens created by mutations in the patient's tumor would be unlikely to be represented in an allogeneic vaccine. Although allogeneic vaccines can be manufactured in sufficient quantities to allow large-scale trials, there remain significant limiting issues in the manufacture and standardization of the vaccine product.
Innovations and Challenges in Melanoma: Summary Statement from the First Cambridge Conference
Authors: Atkins Michael B. , Elder David E. , Essner Richard , Flaherty Keith T. , Gajewski Thomas F. , Haluska Frank G. , Hwu Patrick , Keilholz Ulrich , Kirkwood John M. , Mier James W. , Ross Merrick I. , Slingluff Craig L. , Sondak Vernon K. , Sosman Jeffrey A. , Weinstock Martin A. , King Laura
Published: April 2006

Innovations and Challenges in Melanoma, chaired by Michael Atkins and cochaired by Ulrich Keilholz, John Kirkwood, and Jeffrey Sosman, was held July 15 to 16, 2005, in Cambridge, Massachusetts. The conference brought together leading experts in the fields of cancer research, medical oncology, surgical oncology, anatomic pathology, dermatology, and immunotherapy who wished to advance the field of melanoma treatment by exchanging information and perspectives regarding recent advances and recommendations for further study. The conference proceedings published in this educational supplement to Clinical Cancer Research are intended to provide timely information and recommendations on how genetics, biology, and data information can enhance our understanding of melanoma biology and help inform the use of therapies for this disease.
Reversal of the Malignant Phenotype of Cervical Cancer CaSki Cells through Adeno-Associated Virus–Mediated Delivery of HPV16 E7 Antisense RNA
Authors: Wu Sufang , Meng Li , Wang Shixuan , Wang Wei , Xi Ling , Tian Xun , Chen Gang , Wu Ying , Zhou Jianfeng , Xu Gang , Lu Yunping , Ma Ding
Published: April 2006

Human papillomavirus (HPV) infection is the most important risk factor for the development of cervical cancer. The oncogene E7 from high-risk HPV strains has the ability to immortalize epithelial cells and increase cellular transformation in culture. In this study, we explored the possibility of preventing cervical cancer growth by inhibiting HPV16 E7 expression through gene transfer of an antisense construct. A recombinant adeno-associated virus (rAAV) vector was chosen for the transfer, based on its transfection efficiency, in vivo stability, and lack of detectable pathology. In vitro transfer of an rAAV vector expressing antisense HPV16 E7 (AAV-HPV16E7AS) inhibited cell proliferation, induced apoptosis, reduced cell migration, and restrained in vivo proliferation of HPV16/HPV18–positive cervical cancer CaSki cells. These results indicate that down-regulation of HPV16 E7 with antisense RNA is beneficial in reducing the tumorigenicity of CaSki cells, and rAAV vectors ought to be a new efficient approach for delivering the expression of therapeutic genes.
Prevention of Ultraviolet Radiation–Induced Immunosuppression by (−)-Epigallocatechin-3-Gallate in Mice Is Mediated through Interleukin 12–Dependent DNA Repair
Authors: Meeran Syed M. , Mantena Sudheer K. , Katiyar Santosh K.
Published: April 2006

Purpose: Solar UV radiation–induced immunosuppression is considered to be a risk factor for melanoma and nonmelanoma skin cancers. We previously have shown that topical application of (−)-epigallocatechin-3-gallate (EGCG) prevents UV-induced immunosuppression in mice. We studied whether prevention of UV-induced immunosuppression by EGCG is mediated through interleukin 12 (IL-12)–dependent DNA repair.

Experimental Design: IL-12 knockout (KO) mice on C3H/HeN background and DNA repair–deficient cells from xeroderma pigmentosum complementation group A (XPA) patients were used in this study. The effect of EGCG was determined on UV-induced suppression of contact hypersensitivity and UV-induced DNA damage in the form of cyclobutane pyrimidine dimers (CPD) in mice and XPA-deficient cells using immunohistochemistry and dot-blot analysis.

Results: Topical treatment with EGCG prevented UV-induced suppression of the contact hypersensitivity in wild-type (WT) mice but did not prevent it in IL-12 KO mice. Injection of anti-IL-12 monoclonal antibody to WT mice blocked the preventive effect of EGCG on UV-induced immunosuppression. EGCG reduced or repaired UV-induced DNA damage in skin faster in WT mice as shown by reduced number of CPDs+ cells and reduced the migration of CPD+ antigen-presenting cells from the skin to draining lymph nodes. In contrast, this effect of EGCG was not seen in IL-12 KO mice. Further, EGCG was able to repair UV-induced CPDs in XPA-proficient cells obtained from healthy person but did not repair in XPA-deficient cells, indicating that nucleotide excision repair mechanism is involved in DNA repair.

Conclusions: These data identify a new mechanism by which EGCG prevents UV-induced immunosuppression, and this may contribute to the chemopreventive activity of EGCG in prevention of photocarcinogenesis.
Glycoprotein Nonmetastatic Melanoma Protein B, a Potential Molecular Therapeutic Target in Patients with Glioblastoma Multiforme
Authors: Kuan Chien-Tsun , Wakiya Kenji , Dowell Jeannette M. , Herndon James E. , Reardon David A. , Graner Michael W. , Riggins Gregory J. , Wikstrand Carol J. , Bigner Darell D.
Published: April 2006

Purpose: More brain tumor markers are required for prognosis and targeted therapy. We have identified and validated promising molecular therapeutic glioblastoma multiforme (GBM) targets: human transmembrane glycoprotein nonmetastatic melanoma protein B (GPNMBwt) and a splice variant form (GPNMBsv, a 12-amino-acid in-frame insertion in the extracellular domain).

Experimental Design: We have done genetic and immunohistochemical evaluation of human GBM to determine incidence, distribution, and pattern of localization of GPNMB antigens in brain tumors as well as survival analyses.

Results: Quantitative real-time PCR on 50 newly diagnosed GBM patient tumor samples indicated that 35 of 50 GBMs (70%) were positive for GPNMBwt+sv transcripts and 15 of 50 GBMs (30%) were positive for GPNMBsv transcripts. Normal brain samples expressed little or no GPNMB mRNA. We have isolated and characterized an anti-GPNMB polyclonal rabbit antiserum (2640) and two IgG2b monoclonal antibodies (mAb; G11 and U2). The binding affinity constants of the mAbs ranged from 0.27 × 108 to 9.6 × 108 M−1 measured by surface plasmon resonance with immobilized GPNMB, or 1.7 to 2.1 × 108 M−1 by Scatchard analyses with cell-expressed GPNMB. Immunohistochemical analysis detected GPNMB in a membranous and cytoplasmic pattern in 52 of 79 GBMs (66%), with focal perivascular reactivity in ∼27%. Quantitative flow cytometric analysis revealed GPNMB cell surface molecular density of 1.1 × 104 to 7.8 × 104 molecules per cell, levels sufficient for mAb targeting. Increased GPNMB mRNA levels correlated with elevated GPNMB protein expression in GBM biopsy samples. Univariate and multivariate analyses correlated expression of GPNMB with survival of 39 GBM patients using RNA expression and immunohistochemical data, establishing that patients with relatively high mRNA GPNMB transcript levels (wt+sv and wt), >3-fold over normal brain, as well as positive immunohistochemistry, have a significantly higher risk of death (hazard ratios, 3.0, 2.2, and 2.8, respectively).

Conclusions: Increased mRNA and protein levels in GBM patient biopsy samples correlated with higher survival risk; as a detectable surface membrane protein in glioma cells, the data indicate that GPNMB is a potentially useful tumor-associated antigen and prognostic predictor for therapeutic approaches with malignant gliomas or any malignant tumor that expresses GPNMB.
Cross Talk Between Endothelial and Smooth Muscle Cells in Pulmonary Hypertension
Authors: Eddahibi Saadia , Guignabert Christophe , Barlier-Mur Anne-Marie , Dewachter Laurence , Fadel Elie , Dartevelle Philippe , Humbert Marc , Simonneau Gerald , Hanoun Naïma , Saurini Françoise , Hamon Michel , Adnot Serge
Published: April 2006

Background—

The mechanism of pulmonary artery smooth muscle cell (PA-SMC) hyperplasia in idiopathic pulmonary artery hypertension (iPH) may involve both an inherent characteristic of PA-SMCs and abnormal control by external stimuli. We investigated the role of pulmonary microvascular endothelial cells (P-ECs) in controlling PA-SMC growth.

Methods and Results—

Serum-free medium of quiescent P-ECs elicited marked PA-SMC proliferation, and this effect was greater with P-ECs from patients with iPH than from control subjects and greater with PA-SMCs from these patients than from control subjects. Fluoxetine, which inhibits serotonin-induced mitogenesis by blocking the serotonin transporter, and p-chlorophenylalanine, which inhibits serotonin synthesis by blocking tryptophan hydroxylase (TPH), caused a similar 60% reduction in the growth-promoting effect of P-EC media, whereas endothelin receptor blockers had no effect. Assays of TPH activity in P-EC medium based on p-chlorophenylalanine–sensitive 5-hydroxytryptophan accumulation or serotonin determination indicated serotonin synthesis by P-ECs and an increase in this TPH-dependent process in iPH. Expression of the
tph1
gene encoding the peripheral form of the TPH enzyme was increased in lungs and P-ECs from patients with iPH. Lung TPH1 immunostaining was confined to the pulmonary vessel intima.

Conclusions—

P-ECs produce paracrine factors governing PA-SMC growth. Serotonin, the main P-EC–derived growth factor, is overproduced in iPH and contributes to PA-SMC hyperplasia.
Role of Endogenous Fas (CD95/Apo-1) Ligand in Balloon-Induced Apoptosis, Inflammation, and Neointima Formation
Authors: Matter Christian M. , Chadjichristos Christos E. , Meier Patricia , von Lukowicz Tobias , Lohmann Christine , Schuler Pia K. , Zhang Dongming , Odermatt Bernhard , Hofmann Eugen , Brunner Thomas , Kwak Brenda R. , Lüscher Thomas F.
Published: April 2006

Background—

Fas (CD95/Apo-1) ligand (FasL)–induced apoptosis in Fas-bearing cells is critically involved in modulating immune reactions and tissue repair. Apoptosis has also been described after mechanical vascular injury such as percutaneous coronary intervention. However, the relevance of cell death in this context of vascular repair remains unknown.

Methods and Results—

To determine whether FasL-induced apoptosis is causally related to neointimal lesion formation, we subjected FasL-deficient (generalized lymphoproliferative disorder [
gld
], C57BL/6J) and corresponding wild-type (WT) mice to carotid balloon distension injury, which induces marked endothelial denudation and medial cell death. FasL expression in WT mice was induced in injured vessels compared with untreated arteries (
P
<0.05; n=5). Conversely, absence of functional FasL in
gld
mice decreased medial and intimal apoptosis (terminal deoxynucleotidyltransferase–mediated dUTP nick end labeling [TUNEL] index) at 1 hour and 7 days after balloon injury (
P
<0.05; n=6). In addition, peritoneal macrophages isolated from
gld
mice showed no apoptosis and enhanced migration (
P
<0.05; n=4). In parallel, we observed increased balloon-induced macrophage infiltrations (anti-CD68) in injured arteries of FasL-deficient animals (
P
<0.05; n=6). Together with enhanced proliferation (bromodeoxyuridine index;
P
<0.05), these events resulted in a further increase in medial and neointimal cells (
P
<0.01; n=8) with thickened neointima in
gld
mice (intima/media ratio, ×3.8 of WT;
P
<0.01).

Conclusions—

Our data identify proapoptotic and antiinflammatory effects of endogenous FasL as important factors in the process of neointimal lesion formation after balloon injury. Moreover, they suggest that activation of FasL may decrease neointimal thickening after percutaneous coronary intervention.
A Technique to Detect Microclimatic Inhomogeneities in Historical Records of Screen-Level Air Temperature
Authors: Runnalls K. E. , Oke T. R.
Published: April 2006

A new method to detect errors or biases in screen-level air temperature records at standard climate stations is developed and applied. It differs from other methods by being able to detect microclimatic inhomogeneities in time series. Such effects, often quite subtle, are due to alterations in the immediate environment of the station such as changes of vegetation, development (buildings, paving), irrigation, cropping, and even in the maintenance of the site and its instruments. In essence, the technique recognizes two facts: differences of thermal microclimate are enhanced at night, and taking the ratio of the nocturnal cooling at a pair of neighboring stations nullifies thermal changes that occur at larger-than-microclimatic scales. Such ratios are shown to be relatively insensitive to weather conditions. After transforming the time series using Hurst rescaling, which identifies long-term persistence in geophysical phenomena, cooling ratio records show distinct discontinuities, which, when compared against detailed station metadata records, are found to correspond to even minor changes in the station environment. Effects detected by this method are shown to escape detection by current generally accepted techniques. The existence of these microclimatic effects are a source of uncertainty in long-term temperature records, which is in addition to those presently recognized such as local and mesoscale urban development, deforestation, and irrigation.
Simulations of Present and Future Climates in the Western United States with Four Nested Regional Climate Models
Authors: Duffy P. B. , Arritt R. W. , Coquard J. , Gutowski W. , Han J. , Iorio J. , Kim J. , Leung L.-R. , Roads J. , Zeledon E.
Published: April 2006

In this paper, the authors analyze simulations of present and future climates in the western United States performed with four regional climate models (RCMs) nested within two global ocean–atmosphere climate models. The primary goal here is to assess the range of regional climate responses to increased greenhouse gases in available RCM simulations. The four RCMs used different geographical domains, different increased greenhouse gas scenarios for future-climate simulations, and (in some cases) different lateral boundary conditions. For simulations of the present climate, RCM results are compared to observations and to results of the GCM that provided lateral boundary conditions to the RCM. For future-climate (increased greenhouse gas) simulations, RCM results are compared to each other and to results of the driving GCMs. When results are spatially averaged over the western United States, it is found that the results of each RCM closely follow those of the driving GCM in the same region in both present and future climates. This is true even though the study area is in some cases a small fraction of the RCM domain. Precipitation responses predicted by the RCMs in many regions are not statistically significant compared to interannual variability. Where the predicted precipitation responses are statistically significant, they are positive. The models agree that near-surface temperatures will increase, but do not agree on the spatial pattern of this increase. The four RCMs produce very different estimates of water content of snow in the present climate, and of the change in this water content in response to increased greenhouse gases.
Interannual Variability of Indian Ocean Heat Transport
Authors: Chirokova Galina , Webster Peter J.
Published: April 2006

The work in this paper builds upon the relatively well-studied seasonal cycle of the Indian Ocean heat transport by investigating its interannual variability over a 41-yr period (1958–98). An intermediate, two-and-a-half-layer thermodynamically active ocean model with mixed layer physics is used in the investigation. The results of the study reveal that the Indian Ocean heat transport possesses strong variability at all time scales from intraseasonal (10–90 days) to interannual (more than one year). The seasonal cycle dominates the variability at all latitudes, the amplitude of the intraseasonal variability is similar to the seasonal cycle, and the amplitude of the interannual variability is about one-tenth of the seasonal cycle. Spectral analysis shows that a significant broadband biennial component in the interannual variability exists with considerable coherence in sign across the equator. While the mean annual heat transport shows a strong maximum between 10° and 20°S, interannual variability is relatively uniform over a broad latitudinal domain between 15°N and 10°S. The heat transport variability at all time scales is well explained by the Ekman heat transport, with especially good correlations at the intraseasonal time scales. The addition of the Indonesian Throughflow does not significantly affect the heat transport variability in the northern part of the ocean.
A Climatology of Northern Hemisphere Blocking
Authors: Barriopedro David , García-Herrera Ricardo , Lupo Anthony R. , Hernández Emiliano
Published: April 2006

In this paper a 55-yr (1948–2002) Northern Hemisphere blocking climatology is presented. Traditional blocking indices and methodologies are revised and a new blocking detection method is designed. This algorithm detects blocked flows and provides for a better characterization of blocking events with additional information on blocking parameters such as the location of the blocking center, the intensity, and extension. Additionally, a new tracking procedure has been incorporated following simultaneously the individual evolution of blocked flows and identifying coherently persistent blocked patterns.

Using this method, the longest known Northern Hemisphere blocking climatology is obtained and compared with previous studies. A new regional classification into four independent blocking sectors has been obtained based on the seasonally preferred regions of blocking formation: Atlantic (ATL), European (EUR), West Pacific (WPA), and East Pacific (EPA). Global and regional blocking characteristics have been described, examining their variability from the seasonal to interdecadal scales.

The global long-term blocking series in the North Hemisphere showed a significant trend toward weaker and less persistent events, as well as regional increases (decreases) in blocking frequency over the WPA (ATL and EUR) sector. The influence of teleconnection patterns (TCPs) on blocking parameters is also explored, being confined essentially to wintertime, except in the WPA sector. Additionally, regional blocking parameters, especially frequency and duration, are sensitive to regional TCPs, supporting the regional classification obtained in this paper. The ENSO-related blocking variability is evident in blocking intensities and preferred locations but not in frequency. Finally, the dynamical connection between blocking occurrence and regional TCPs is examined through the conceptual model proposed by Charney and DeVore. Observational evidence of a dynamical link between the asymmetrical temperature distributions induced by TCPs and blocking variability is provided with a distinctive contrast “warm ocean/cold land” pattern favoring the blocking occurrence in winter. However, the conceptual model is not coherent in the WPA sector, suggesting different blocking mechanisms operating in this sector.
Assimilation of Directional Wave Spectra in the Wave Model WAM: An Impact Study from Synthetic Observations in Preparation for the SWIMSAT Satellite Mission
Authors: Aouf Lotfi , Lefèvre Jean-Michel , Hauser Danièle
Published: April 2006

Within the framework of the Surface Waves Investigation and Monitoring from Satellite mission (SWIMSAT) proposed to the European Space Agency, an assimilation scheme has been implemented in the Wave Model (WAM) in order to estimate the impact of spectral information on wave prediction. The scheme uses an optimal interpolation and the “spectral partitioning” principle. The synthetic wave spectra are located along a SWIMSAT orbit track and are assimilated in a 4-day-period simulation. Random errors are included to simulate the uncertainties of SWIMSAT instrumentation. The sensitivity of the scheme to background and observational errors and the correlation length is examined. The assimilation impact is investigated for two cases of moderate and large errors of the first guess.

The results show that the assimilation scheme works correctly and the rms errors of significant wave height, mean period, and direction are significantly reduced for both periods of analysis and forecast. The impact on significant wave height is noticeable during the period of analysis and stays efficient for 2-day forecasts. For a large error in the first guess, the impact increases and remains significant for 3-day forecasts.

Statistical analysis of mean wave parameters clearly shows that the use of spectral information yields a better estimate of wave frequency, direction, and low-frequency wave height in comparison with the results based upon assimilation of wave heights only. However, total significant wave height is less sensitive to the addition of spectral information in the assimilation scheme. The use of correlation length depending on the latitude of grid points leads to a better spread of incremental observations and, hence, to better skills in terms of the rms errors of mean wave parameters. The use of several wavelength cutoffs concerning the SWIMSAT synthetic wave spectra suggests that the “assimilation index” of mean wave parameters decreases with the increasing wavelength cutoff.
Organizational discourse and subjectivity
Authors: Bergström Ola , Knights David
Published: April 2006

This article seeks to contribute to the debate on the relationship between organizational discourses and subjectivity, revolving around whether organizational discourses determine individual subjectivity and the extent to which there is room for human agency. It does so by providing empirical illustrations of how organizational discourses constitute subjectivity during processes of recruitment in a large American consultancy firm operating in Sweden. The analysis illustrates how interviewers, by various discursive moves, initiate, support, control and follow up candidates’ decision to join the company, as if it was an independent choice to join. Findings suggest that to the extent that subjectification takes place during the recruitment process it is dependent on the candidate’s use and acceptance of organizational discourses as expressions of their own motives for working at the company. These findings have implications for the understanding of the relationship between organizational discourses and individual subjectivity and how subjectification processes may be studied in other practices and organizations. It argues that subjectification is an effect of the interaction between human agency and organizational discourses rather than in the determination of one to the other. Any attempt to analyse the impact of organizational discourse on individual subjectivity must take into account the possibility that subjects actively take part in their own self-construction and that this construction is produced in social interaction.
A structural model of 20S immunoproteasomes: effect of LMP2 codon 60 polymorphism on expression, activity, intracellular localisation and insight into the regulatory mechanisms
Authors: Mishto Michele , Santoro Aurelia , Bellavista Elena , Sessions Richard , Textoris-Taube Kathrin , Piaz Fabrizio Dal , Carrard Geraldine , Forti Katia , Salvioli Stefano , Friguet Bertrand , Kloetzel Peter M. , Rivett A. Jennifer , Franceschi Claudio
Published: April 2006

The immunoproteasome subunit low molecular weight protein 2 (LMP2) codon 60 polymorphism has been associated with autoimmune diseases. It has also been demonstrated to influence susceptibility to TNF-α-induced apoptosis in blood cells and proteasome activity in aged human brain. In the present study, anin silicomodel of immunoproteasome was used to examine the effect of the R60H polymorphism in the LMP2 subunit. The investigation of immunoproteasome expression, activity and intracellular localisation in anin vitrocellular model, namely lymphoblastoid cell lines, showed no major variations in functionality and amount, while a significant difference in antibody affinity was apparent. These data were integrated with previous results obtained in different tissues and combined with a structural model of the LMP2 polymorphism. Accordingly, we identified three prospective mechanisms that could explain the biological data for the polymorphism, such as modulation of the binding affinity of a putative non-catalytic modifier site on the external surface of the immunoproteasome core, or the modification of any channel between α and β rings.
Monomeric and dimeric GDF-5 show equal type I receptor binding and oligomerization capability and have the same biological activity
Authors: Sieber Christina , Plöger Frank , Schwappacher Raphaela , Bechtold Rolf , Hanke Michael , Kawai Shinji , Muraki Yoshifumi , Katsuura Mieko , Kimura Michio , Rechtman Maya Mouler , Henis Yoav I. , Pohl Jens , Knaus Petra
Published: April 2006

Growth and differentiation factor 5 (GDF-5) is a homodimeric protein stabilized by a single disulfide bridge between cysteine 465 in the respective monomers, as well as by three intramolecular cysteine bridges within each subunit. A mature recombinant human GDF-5 variant with cysteine 465 replaced by alanine (rhGDF-5 C465A) was expressed inE. coli, purified to homogeneity, and chemically renatured. Biochemical analysis showed that this procedure eliminated the sole interchain disulfide bond. Surprisingly, the monomeric variant of rhGDF-5 is as potentin vitroas the dimeric form. This could be confirmed by alkaline phosphatase assays and Smad reporter gene activation. Furthermore, dimeric and monomeric rhGDF-5 show comparable binding to their specific type I receptor, BRIb. Studies on living cells showed that both the dimeric and monomeric rhGDF-5 induce homomeric BRIb and heteromeric BRIb/BRII oligomers. Our results suggest that rhGDF-5 C465A has the same biological activity as rhGDF-5 with respect to binding to, oligomerization of and signaling through the BMP receptor type Ib.
Messsystem zur nichtinvasiven Blutdruckerfassung. Störunterdrückung in biomedizinischen Signalen mittels Multi-Skalen-Analyse (Non-Invasive Measurement of the Blood Pressure. Noise Reduction in Biomedical Signals Using Multi-Resolution Analysis)
Authors: Wack Achim , Koch Alexander W.
Published: April 2006

Dieser Artikel befasst sich mit der nichtinvasiven Messung des peripheren Blutdruckes beim Menschen unter Belastung, beispielsweise auf einem Laufband. Dabei treten in der Regel so starke Artefakte auf, dass aufgrund eines sehr geringen SNR eine Messung mit herkömmlichen Geräten nicht möglich ist. Mittels einer speziellen Multi-Skalen-Analyse konnte das SNR signifikant verbessert werden.