bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in the quaternary structure of amelogenin when adsorbed onto surfaces
Authors: Tarasevich Barbara J. , Lea Scott , Bernt William , Engelhard Mark H. , Shaw Wendy J.
Published: September 2008

Amelogenin is a unique protein that self‐assembles into spherical aggregates called “nanospheres” and is believed to be involved in controlling the formation of the highly anisotropic and ordered hydroxyapatite crystallites that form enamel. The adsorption behavior of amelogenin onto substrates is of great interest because protein‐surface interactions are critical to its function. We report studies of the adsorption of amelogenin onto self‐assembled monolayers containing COOH end group functionality as well as single crystal fluoroapatite, a biologically relevant surface. We found that although our solutions contained only nanospheres of narrow size distribution, smaller structures such as dimers or trimers were observed on the hydrophilic surfaces. This suggests that amelogenin can adsorb onto surfaces as small structures that “shed” or disassemble from the nanospheres that are present in solution. © 2008 Wiley Periodicals, Inc. Biopolymers 91: 103–107, 2009.

This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com
Design and synthesis of backbone cyclic phosphorylated peptides: The IκB model
Authors: Qvit Nir , Hatzubai Ada , Shalev Deborah E. , Friedler Assaf , Ben‐Neriah Yinon , Gilon Chaim
Published: September 2008

Phosphopeptides have been used to study phosphorylation and dephosphorylation, which are key events in protein expression. Backbone cyclization has been shown to increase the stability and selectivity of peptides. Backbone cyclic peptides with conformational diversity have produced bioactive peptides with improved pharmaceutical properties, metabolic stability, and enhanced intestinal permeability. We demonstrate a successful methodology for incorporating phospho‐amino acids into backbone cyclic peptides. The nuclear factor‐kappa B (NF‐κB) is a latent mammalian protein prototype of dimeric transcription factors that exists in all cell types and plays a pivotal role in a huge number of genes, such as those responsible for chronic and acute inflammatory diseases. To inhibit NF‐κB, backbone cyclic phosphopeptides were designed and synthesized based on the conserved sequence of the Inhibitor kappa B (IκB). The peptides were screened for inhibiting IκB ubiquitylation. The best compound showed 90% inhibition at a concentration of 3 μM, and its solution structure showed similarity to a related β‐catenin protein. This general methodology can be use for synthesizing cyclic phosphorylated, as well as backbone cyclic phosphorylated peptides for various biological targets. © 2008 Wiley Periodicals, Inc. Biopolymers 91: 157–168, 2009.

This article was originally published online as an accepted preprint. The “Published Online“ date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com
Wavelength selection using the measure of topological relevance on the self‐organizing map
Authors: Corona Francesco , Reinikainen Satu‐Pia , Aaljoki Kari , Perkiö Annikki , Liitiäinen Elia , Baratti Roberto , Simula Olli , Lendasse Amaury
Published: September 2008

In this work, we investigated the possibility to perform wavelength selection by exploiting the metric structure of the spectrophotoscopic measurements. The topologically preserving representation of the data is performed using the self‐organizing map (SOM) where the inputs' significance to the output is computed with the measure of topological relevance (MTR) on SOM. The MTR on SOM is a metric measuring the similarity between local distance matrices and we found that spectral inputs with a topology, which is, close to the output's are also associated to the wavelengths that chemically explain the influence of the spectra to the property of interest. As a result, we suggest a wavelength selection strategy based on the MTR on SOM, that is, interpretable to the domain experts and independent on the regression technique subsequently used for estimation. To support the presentation, a full‐scale application from the oil refining industry is illustrated on the problem of estimating standard properties in a complex hydrocarbon product starting from spectrophotoscopic measurements. The method is further validated on the problem of octane number estimation in finished gasolines, under small sample conditions. The application led to accurate, parsimonious and understandable models. Copyright © 2008 John Wiley & Sons, Ltd.
Detection and quantification of (R) and (S)‐dechloroethylifosfamide metabolites in plasma from children by enantioselective LC/MS/MS
Authors: Aleksa Katarina , Nava‐Ocampo Alejandro , Koren Gideon
Published: September 2008

Ifosfamide (IF), a potent chemotherapeutic agent for solid tumors, is known to cause high rates of nephrotoxicity in children with cancer, which is most likely due to the renal production of the metabolite chloroacetaldehyde. Using plasma samples obtained from pediatric oncology patients, we developed a simple nonderivatizing enantioselective liquid chromatography mass spectrometry method to detect the (R) and (S)‐2‐ and 3‐dechloroethylifosfamide metabolites. The (R) and (S)‐enantiomers of the 2‐ and 3‐DCEIF (N‐3‐dechlroethylifosfamide) were detectable in all 22 patients' samples with levels ranging from 9.9 to 238.7 ng/ml for (R)‐2‐DCEIF, 15.8 to 663.0 ng/ml for (S)‐2‐DCEIF, 20.8 to 852.8 ng/l for (R)‐3‐DCEIF and 28.0 to 862.0 ng/ml for (S)‐3‐DCEIF. In addition, the lower limit of quantification for this method is 1 ng/ml. Future studies should concentrate on (R) or (S) production of the 2‐DCEIF and 3‐DCEIF and subsequently chloroacetaldehyde formation with the aim of considering the administration of only the (R)‐IF as its metabolism results in a lower production of chloroacetaldehyde. Chirality, 2009. © 2008 Wiley‐Liss, Inc.
A new method for the detection of microorganisms in blood cultures: Part I. Theoretical analysis and simulation of blood culture processes
Authors: Smith Jennifer M. , Serebrennikova Yulia M. , Huffman Debra E. , Leparc German F. , García‐Rubio Luis H.
Published: September 2008

The physical and chemical changes occurring in blood as a consequence of microbial activity can be used as quantitative indicators of the presence of microorganisms in blood cultures. This paper reports on the theoretical analysis and computer simulation of the changes in the physical and chemical properties of blood expected as a result of the presence of microorganisms and explores the possibilities of spectrophotometric systems for the early detection of pathogens. It is concluded that multi‐wavelength reflectance methods are suitable and that the approach reported herein can lead to considerable simplifications and cost reduction of blood culture systems.
Control relevant on‐line model validation criterion based on robust stability conditions
Authors: Jiang Hailei , Shah Sirish L. , Huang Biao
Published: September 2008

This paper is concerned with detection and diagnosis of modelling error under closed‐loop conditions. The effect of modelling error on process output error (which is the error between the process output and the simulated output) is first analysed. Then robust stability conditions for on‐line model validation are applied. The main premise is that whenever the closed‐loop system violates the robust stability condition, it is a sign of significant process change and a signal that the control system may become potentially unstable. We relate the process output error with robust stability conditions and introduce three propositions for on‐line model validation. Any process change (or modelling error) that makes the system violate the condition specified by the robust stability theorem can be detected. Simulation examples are presented to demonstrate the applicability of the propositions. An index is also proposed to quantify modelling error in frequency domain. Simulation examples and an experimental case study are presented to demonstrate the use of the new index.
Neoadjuvant gemcitabine and cisplatin: Another step on the path toward improving the outcomes of patients with high‐risk, invasive bladder cancer
Authors: Nielsen Matthew E.
Published: September 2008

Until such time as there are data from prospective randomized trials directly comparing neoadjuvant cisplatin and gemcitabine (GC) to neoadjuvant methotrexate, vinblastine, doxorubicin, and cisplatin, the data presented in the study by Dash et al in this issue of Cancer provide strong support for the consideration of neoadjuvant GC as an effective and well‐tolerated component in the armamentarium against high‐risk bladder cancer.
Cancer‐specific survival outcomes among patients treated during the cytokine era of kidney cancer (1989‐2005)
Authors: Belldegrun Arie S. , Klatte Tobias , Shuch Brian , LaRochelle Jeffrey C. , Miller David C. , Said Jonathan W. , Riggs Stephen B. , Zomorodian Nazy , Kabbinavar Fairooz F. , deKernion Jean B. , Pantuck Allan J.
Published: September 2008

BACKGROUND.

The management of renal cell carcinoma (RCC) is evolving toward less extirpative surgery and the use of targeted therapy. The authors set out to provide a benchmark against which emerging therapies should be measured.

METHODS.

A prospective database including clinical and pathological variables for 1632 patients with RCC treated between 1989 and 2005 was queried. Patients were stratified using the University of California‐Los Angeles Integrated Staging System (UISS) into low‐, intermediate‐, and high‐risk groups. Disease‐specific survival (DSS) was measured. Response to systemic therapy for patients with advanced disease was assessed.

RESULTS.

Nephrectomy was performed in 1492 patients. Overall 5‐, 10‐, and 15‐year DSS was 55%, 40%, and 29%. For localized disease, 5‐ and 10‐year DSS for UISS low‐, intermediate‐, and high‐risk groups was 97% and 92%, 81% and 61%, and 62% and 41%, respectively. For metastatic disease, 5‐ and 10‐year DSS for UISS low‐, intermediate‐, and high‐risk groups was 41% and 31%, 18% and 7%, and 8% and 0%, respectively. Patients with metastatic disease receiving immunotherapy (n = 453) had complete response in 7% (median survival [MS], 120+ months), partial response in 15% (MS, 42.8 months), stable disease in 33% (MS, 38.6 months), and progressive disease in 45% (MS, 11.6 months).

CONCLUSIONS.

Most patients with localized RCC do well with surgery alone, but effective adjuvant therapy is needed for patients identified as at high risk for recurrence. For advanced disease, newer targeted and potentially less toxic treatments should be at least as effective as those achieved with aggressive surgical resection and immunotherapy. Cancer 2008. © 2008 American Cancer Society.
Short‐term effects of treatment‐induced hormonal changes on cognitive function in breast cancer patients
Authors: Hermelink Kerstin , Henschel Volkmar , Untch Michael , Bauerfeind Ingo , Lux Michael Patrick , Munzel Karin
Published: September 2008

BACKGROUND.

It is suspected that estrogen depletion resulting from treatment may contribute to cognitive compromise in patients with breast cancer. However, the evidence for estrogen effects on cognition is inconclusive, and the consequences of hormonal changes for cognitive function in patients with cancer rarely have been investigated. In this study, the authors investigated the effects of treatment‐induced menopause and antiestrogen therapy with tamoxifen and aromatase inhibitors (AIs) on cognitive function.

METHODS.

Cognitive performance was assessed in 101 patients with breast cancer before the start of cancer therapy (T1), toward the end of neoadjuvant chemotherapy (T2), and 1 year after baseline (T3) using 12 cognitive tests. Menopause occurred in a subgroup of patients, and an overlapping subgroup started antiestrogen therapy with tamoxifen or AIs. Linear mixed‐effects models that made it possible to determine effects at group levels and individual levels simultaneously were used for statistical analysis.

RESULTS.

At the group level, a significant favorable effect of induced menopause emerged in a test of executive function (P = .0035). Two additional group‐level effects of induced menopause, both favorable, and 2 individual‐level effects that were positive in some patients and negative in others were not significant when multiple testing was taken into account. No significant effects of tamoxifen or AIs on cognitive function were observed.

CONCLUSIONS.

Hormonal changes did not appear to contribute to cognitive compromise in patients with breast cancer during the first year after diagnosis. Antiestrogen treatment with tamoxifen or AIs did not affect cognition, and the effects of induced menopause were more likely to be favorable. However, the possibility that some cognitive decline occurs in individual patients could not be excluded. Cancer 2008. © 2008 American Cancer Society.
Genetic variations in cell‐cycle pathway and the risk of oral premalignant lesions
Authors: Ye Yuanqing , Lippman Scott M. , Lee J. Jack , Chen Meng , Frazier Marsha L. , Spitz Margaret R. , Wu Xifeng
Published: September 2008

BACKGROUND.

Cell‐cycle checkpoint controls regulate cell‐cycle progression and proliferation. Alterations in cell‐cycle control mechanisms are linked to tumorigenesis.

METHODS.

This case‐control study included 147 cases and 147 controls. The authors used a pathway‐based approach to assess the association between 10 potential functional single‐nucleotide polymorphisms from 7 cell‐cycle control genes and the risk of oral premalignant lesions (OPLs). They also used classification and regression tree analysis to examine high‐order gene‐gene and gene‐smoking interactions.

RESULTS.

Compared with the homozygous wild‐type GG genotype of CCND1 P241P, individuals with the AG genotype exhibited an increased risk of OPL (odds ratio, 1.58; 95% confidence interval, 0.89‐2.83), and carriers of the AA genotype had a significantly increased risk of OPL (odds ratio, 2.75; 95% confidence interval, 1.33‐5.71), with risk increasing significantly with the increasing number of variant alleles (P = .006). The risk of OPL increased significantly as the number of unfavorable genotypes in the pathway increased (P = .002). The final decision tree in the classification and regression tree analysis contained 5 terminal nodes. Compared with the never smokers (the lowest risk group), the odds ratios for terminal nodes 2 through 5 ranged from 1.21 to 5.40.

CONCLUSIONS.

The results illustrated the advantage of using a pathway‐based approach for analyzing gene‐gene and gene‐smoking interactions. Specifically, the authors showed that genetic polymorphisms in cell‐cycle control pathway genes may contribute to the risk of OPL. Cancer 2008. © 2008 American Cancer Society.
Risk of radiation‐related salivary gland carcinomas among survivors of Hodgkin lymphoma
Authors: Boukheris Houda , Ron Elaine , Dores Graça M. , Stovall Marilyn , Smith Susan A. , Curtis Rochelle E.
Published: September 2008

BACKGROUND.

Radiotherapy for Hodgkin lymphoma (HL) increases the risk of salivary gland carcinomas (SGC). To the authors' knowledge, however, the magnitude of the risk has not been assessed to date.

METHODS.

The risks of SGC among 20,928 1‐year survivors of HL who were diagnosed between 1973 and 2003 were evaluated in 11 population‐based cancer registry areas of the Surveillance, Epidemiology, and End Results (SEER) program. Observed‐to‐expected ratios (O/E) were assessed by radiation treatment, sex, age at the time of HL diagnosis, calendar year of diagnosis, attained age, time since HL diagnosis, histologic type of SGC, and site of occurrence in the major salivary glands.

RESULTS.

Among 11,047 HL patients who received radiotherapy as part of their initial treatment for HL, 21 developed subsequent invasive SGC (O/E = 16.9; 95% confidence interval [95% CI], 10.4‐25.8). The risk of radiation‐related SGC was highest for younger HL patients (age <20 years) (O/E = 45.5; 95% CI, 12.4‐116.5) and among 10‐year survivors (O/E = 23.9; 95% CI, 13.1‐40.1), with risks remaining elevated for at least 2 decades after irradiation. Significant differences in risk by histologic type were observed, with a particularly high risk of developing mucoepidermoid carcinomas (O = 14; O/E = 44.2 [95% CI, 24.2‐74.2]) and adenocarcinomas (O = 4; O/E = 30.6 [95% CI, 8.3‐78.2]) noted.

CONCLUSIONS.

HL patients treated with radiotherapy experienced a significantly increased risk of SGC, particularly when exposed at young ages or for at least 2 decades after exposure. Although the results of the current study reflect the late effects of former HL treatment approaches, they point to the importance of long‐term follow‐up and a heightened awareness of SGC risk in this population. Cancer 2008. Published 2008 by the American Cancer Society.
CD4 and CD8 enumeration for HIV monitoring in resource‐constrained settings
Authors: Li Xiao , Breukers Christian , Ymeti Aurel , Lunter Björn , Terstappen Leon W. M. M. , Greve Jan
Published: September 2008

Background:

We developed a volumetric single platform image cytometer (SP ICM) that is dedicated to count CD4+ and CD8+ T lymphocytes for HIV monitoring in resource‐constrained settings. The instrument was designed to be low‐cost, yet reliable, easy‐to‐use, and robust.

Methods:

Whole blood is incubated with CD3‐magnetic nanoparticles, CD4‐phycoerythrin (PE), and CD8‐peridinin‐chlorophyll‐protein complex (PerCP). The CD3 cells are immunomagnetically attracted to an analysis surface, where fluorescence images of CD4+ and CD8+ T lymphocytes are recorded and analyzed, respectively. We compared CD4, CD8 counts, and CD4/CD8 ratio obtained by the SP ICM with those from a SP flow cytometer (FCM) tetraCXP method on blood samples from 145 patients.

Results:

Good correlations were obtained (R: 0.96–0.99) between the SP ICM and the SP FCM. There was ∼10% CD8 undercount in the SP ICM, which could be partly caused by CD8+dim T lymphocytes that were not detected by the instrument or not counted by the image analysis due to the cross‐talk from the CD4‐PE signal in the CD8‐PerCP image.

Conclusions:

The SP ICM is a good candidate for HIV monitoring in point‐of‐care settings of resource‐constrained countries. © 2008 Clinical Cytometry Society
A comparison of multiplex suspension array large‐panel kits for profiling cytokines and chemokines in rheumatoid arthritis patients
Authors: Khan Imran H. , Krishnan V. V. , Ziman Melanie , Janatpour Kim , Wun Ted , Luciw Paul A. , Tuscano Joseph
Published: September 2008

Background

Multiplex analysis allows measurements of a large number of analytes simultaneously in each sample. On the basis of the Luminex multiplex technology (xMAP), kits for measuring multiple cytokines and chemokines (immunomodulators) are commercially available and are useful in investigations on inflammatory diseases. This study evaluated four multiplex kits (Bio‐Plex, LINCOplex, Fluorokine, and Beadlyte) that contained 27, 29, 20, and 22 analytes each, respectively, for the analysis of immunomodulators in plasma of patients with rheumatoid arthritis (RA) who underwent treatment with antibody against CD20 (rituximab), a B‐cell reductive therapy.

Methods

Multiplex kits were tested on serial plasma samples obtained from six RA patients at baseline and multiple time points (3, 6, and 9 months) post‐treatment with rituximab. The RA patients included in this study had previously failed therapy with disease modifying anti‐arthritis drugs (DMARD) and treatment with anti‐TNFα antibody (infliximab).

Results

Computer modeling and hierarchical cluster analysis of the multiplex data allowed a comparison of the performance of multiplex assay kits and revealed profiles of immunomodulators in the RA patients.

Conclusions

In plasma of RA patients who appeared to have benefited from the rituximab treatment, the profile of significantly elevated immunomodulators by at least two of the three kits (BioPlex, LINCOplex, Beadlyte) is as follows: IL‐12p70, Eotaxin, IL‐4, TNFα, Il‐9, IL‐1β, IFNγ, IL‐10, IL‐6, and IL‐13. Immunomodulator profiling by multiplex analysis may provide useful plasma biomarkers for monitoring response to B‐cell reductive therapy in RA patients. © 2008 Clinical Cytometry Society
A rapid, fluorescence‐based assay for detecting antigenic modulation of the acetylcholine receptor on human cell lines
Authors: Keefe Dennis , Hess Donna , Bosco Jennifer , Tzartos Socrates , Powell Jan , Lamsa Justin , Josiah Serene
Published: September 2008

Background:

Myasthenia gravis (MG) is an autoimmune disease affecting ∼40,000 patients in the United States. One of the major mechanisms of disease pathology in MG is the binding, internalization, and eventual destruction of acetylcholine receptors (AChR) at the neuromuscular junction by cross‐linking AChR‐specific autoantibodies. This process, known as antigenic modulation, ultimately attenuates the ability of muscle cells to contract in response to signals from neurons, leading to muscle weakness and fatigue. For this reason, antigenic modulation of the AChR on cultured cells has become an important diagnostic tool for assessing the pathogenicity of AChR‐specific autoantibodies. Traditionally, these assays have been done using radiolabeled AChR ligands such as 125I alpha‐bungarotoxin to determine relative AChR number. Here, we present a high‐throughput immunofluorescent flow cytometry‐based assay that can be used to quantify AChR levels on the cell surface and assess the efficacy of molecules designed to rescue antigenic modulation.

Methods:

AChR levels were quantified on human muscle cells before and after treatment with AChR antibodies via immunofluorescent labeling with the AChR monoclonal antibodies, mAb210 and mAb B3, followed by flow cytometry of EDTA‐treated cells.

Results:

Using a novel, flow cytometry‐based assay, antigenic modulation of the AChR was demonstrated on human cells using both AChR‐specific monoclonal antibody and MG patient serum. The degree of antigenic modulation was dose responsive to antibody levels and could be reversed by preincubating antibodies with soluble AChR alpha subunit extracellular domain.

A rapid, nonradioactive assay was developed to determine the potential of AChR‐specific antibodies in the serum of MG patients to bind and down‐regulate the AChR. This assay can be used to assess the ability of putative therapeutics that rescue antigenic modulation and could be developed for the treatment of MG. © 2008 Clinical Cytometry Society
PD‐1 ligands expressed on myeloid‐derived APC in the CNS regulate T‐cell responses in EAE
Authors: Schreiner Bettina , Bailey Samantha L. , Shin Tahiro , Chen Lieping , Miller Stephen D.
Published: September 2008

Disease progression in experimental autoimmune encephalomyelitis (EAE) is regulated by programmed death receptor 1 (PD‐1) and its ligands, B7‐H1 (programmed death ligand 1 (PD‐L1)) and B7‐DC (PD‐L2). B7‐H1 and B7‐DC have negative regulatory effects upon binding PD‐1 on activated T cells and B7‐H1 deficiency increases severity of both diabetes and EAE. However, the role of PD‐L expression on different APC in the CNS in regulating local T‐cell function during relapsing EAE has not been examined. Our data show that the majority of CNS CD4+ T cells isolated during acute EAE are PD‐1+, and T cells specific for relapse‐associated epitopes express PD‐1 upon antigen stimulation in the CNS. B7‐H1 and B7‐DC are differentially expressed on discrete APC populations in the inflamed CNS. B7‐H1 and PD‐1 have mainly inhibitory functions on CNS T cells. B7‐H1 negatively regulates the stimulation of activated PD‐1+ TH cells, in co‐cultures with microglia and different CNS‐infiltrating APC presenting endogenously processed peptides. The preponderance of IFN‐γ+ versus IL‐17+ T cells in the CNS of B7‐H1−/− mice suggests that B7‐H1 more selectively suppresses TH‐1 than TH‐17 responses in vivo. In contrast, blockade of B7‐DC has less pronounced regulatory effects. Overall, the results demonstrate that B7‐H1 expressed by CNS myeloid APC negatively regulates T‐cell activation during acute relapsing EAE.
Collagen type II is recognized by a pathogenic antibody through germline encoded structures
Authors: Böiers Ulrika , Lanig Harald , Sehnert Bettina , Holmdahl Rikard , Burkhardt Harald
Published: September 2008

Collagen type II (CII) is a cartilage‐specific target of pathologic humoral autoimmune responses in rheumatoid arthritis as well as in the collagen‐induced arthritis model. The aim of the present study is to investigate the critical amino acid residues conferring CII epitope specificity of the prototypic arthritogenic murine mAb CIIC1. A homology model of the CIIC1 single‐chain antibody fragment (CIIC1scFv) in complex with its triple helical epitope was established. In silico predictions based on extensive molecular dynamics simulations were experimentally tested by the recombinant expression and functional analysis of CIIC1scFv containing alanine replacements allowing the identification of crucial CII‐binding sites in the CDR2 and CDR3 regions of both heavy and light chains. Since the conversion of the CIIC1scFv sequence into the respective germline at all 13 somatically mutated positions did not affect its CII binding, our data indicate that potentially harmful cartilage‐specific humoral autoimmunity could be germline encoded. The molecular modeling further demonstrates that the rigid collagen triple helix restricts the likelihood of molecular interactions with the CDR regions of the antibody considerably compared with globular antigens. These sterical constraints provide an explanation as to why somatic mutations in the arthritogenic autoantibody have no obvious impact on CII recognition.
The capacity of the TNF family members 4‐1BBL, OX40L, CD70, GITRL, CD30L and LIGHT to costimulate human T cells
Authors: Kober Johanna , Leitner Judith , Klauser Christoph , Woitek Ramona , Majdic Otto , Stöckl Johannes , Herndler‐Brandstetter Dietmar , Grubeck‐Loebenstein Beatrix , Reipert Birgit M. , Pickl Winfried F. , Pfistershammer Katharina , Steinberger Peter
Published: September 2008

Activating signals generated by members of the tumour necrosis factor receptor superfamily upon interaction with their cognate ligands play important roles in T‐cell responses. Members of the tumour necrosis factor family namely 4‐1BBL, OX40L, CD70, GITRL, LIGHT and CD30L have been described to function as costimulatory molecules by binding such receptors on T cells. Using our recently described system of T‐cell stimulator cells we have performed the first study where all these molecules have been assessed and compared regarding their capacity to costimulate proliferation and cytokine production of human T cells. 4‐1BBL, which we found to be the most potent molecule in this group, was able to mediate sustained activation and proliferation of human T cells. OX40L and CD70 were also strong inducers of T‐cell proliferation, whereas the costimulatory capacity of human GITRL was significantly lower. Importantly CD30L and LIGHT consistently failed to act costimulatory on human T cells, and we therefore suggest that these molecules might be functionally distinct from the costimulatory members of this family.
Freeze‐and‐thaw‐disrupted tumour cells impair the responsiveness of DC to TLR stimulation
Authors: Tirapu Iñigo , Lewis Amy , Kreutz Martin , McLinden Hayley , Diebold Sandra S.
Published: September 2008

Cancer immunotherapy aims at inducing immune responses against tumour‐associated antigens that mediate the eradication of tumour cells. For successful vaccination against antigens expressed by the tumour, the immune system has to be provided with sufficient amounts of these antigens in connection with strong immunostimulatory signals such as toll‐like receptor (TLR) ligands. Tumour cells represent a convenient source of relevant tumour‐associated antigens but can have suppressive properties. In this study, we explored how different forms of tumour cell material influence the activation of dendritic cells (DC), which play a crucial role in the induction of anti‐tumour immune responses. We show that freeze‐and‐thaw‐disrupted tumour cells inhibit DC activation in response to TLR stimulation, a phenomenon that is only partially seen with non‐disrupted control cells. This suppression of DC stimulation is independent of tumour cell‐ and species‐specific factors. We tested the hypothesis that phosphatidylserine on cells with disrupted membrane integrity mediates inhibition of TLR‐induced DC activation. Our experimental evidence indicates that phosphatidylserine is not involved in the inhibition of TLR‐mediated DC activation by freeze‐and‐thaw‐disrupted cells. The inhibitory activity associated with disrupted tumour cells could explain why such preparations are less effective tumour vaccines than apoptotic tumour cells.
Efficacy of IL‐2‐ versus IL‐15‐stimulated CD8 T cells in adoptive immunotherapy
Authors: Mueller Katja , Schweier Oliver , Pircher Hanspeter
Published: September 2008

We determined the efficacy of in vitro expanded P14 TCR transgenic CD8 T cells to mediate tumor cell elimination and to protect against viral infection in mice. Contrary to previous studies, an adoptive transfer model without lymphodepletion, vaccination or cytokine treatment was used. Antigen‐activated P14 T cells cultured in IL‐2‐containing medium for 7 days (P14IL‐2) exhibited potent effector cell functions in vitro but did not confer protection against melanoma growth or viral infection. In contrast, P14 T cells cultured in IL‐15 (P14IL‐15) were highly effective in vivo although they displayed only moderate effector functions in vitro. Therapeutic efficacy correlated with the survival of the transferred T cells in the recipients: P14IL‐2 cells disappeared rapidly whereas P14IL‐15 cells persisted for prolonged time. Decreasing the IL‐2 concentration in the culture media improved in vivo survival and efficacy but also lowered the cell yield of the cultures. Finally, we could extend the findings with monoclonal P14 T cells to polyclonal CD8 T cells. Thus, in vitro expansion of antigen‐specific CD8 T cells in IL‐15 allowed the generation of substantial numbers of T cells without inducing terminally differentiated effector cells that turned out to be unfavorable in the transfer model examined here.
Effector CD8+ T cells activated in vitro confer immediate and long‐term tumor protection in vivo
Authors: Perret Rachel , Ronchese Franca
Published: September 2008

We studied the ability of CD8+ T cells activated in vitro to mediate tumor protection after transfer into adoptive hosts. TCR transgenic CD8+ T cells were activated in culture with DC and specific peptide antigen, and briefly expanded in IL‐2 containing medium. Cultured cells acquired a CD44hiCD62Llo phenotype, and following in vivo transfer they preferentially homed to non‐lymphoid tissues and spleen. In vivo, their numbers declined between day 0 and day 20, and then remained relatively stable from day 20 to day 90. Over time, many of the injected cells re‐expressed CD62L, and acquired the ability to localize to secondary lymphoid organs. Transferred T cells underwent low‐level proliferation, expressed IL‐7Rα and IL‐15Rβ, were cytotoxic in vivo, and retained the ability to produce IL‐2, IFN‐γ and TNF‐α upon ex vivo restimulation. In addition, transferred T cells conferred a high degree of tumor protection, which was greatest immediately after injection, and remained significant even when tumor was given 90 days after T‐cell transfer. We conclude that in vitro generated effector T cells can mediate immediate and long‐term tumor protection, and develop into long‐lived memory T‐cell populations in vivo.
DNA polymerase η is a limiting factor for A:T mutations in Ig genes and contributes to antibody affinity maturation
Authors: Masuda Keiji , Ouchida Rika , Yokoi Masayuki , Hanaoka Fumio , Azuma Takachika , Wang Ji‐Yang
Published: September 2008

DNA polymerase η (POLH) is required for the generation of A:T mutations during the somatic hypermutation of Ig genes in germinal center B cells. It remains unclear, however, whether POLH is a limiting factor for A:T mutations and how the absence of POLH might affect antibody affinity maturation. We found that the heterozygous Polh+/− mice exhibited a significant reduction in the frequency of A:T mutations in Ig genes, with each type of base substitutions at a level intermediate between the Polh+/+ and Polh−/− mice. These observations suggest that Polh is haplo‐insufficient for the induction of A:T mutations in Ig genes. Intriguingly, there was also a reduction of C to T and G to A transitions in Polh+/− mice as compared with WT mice. Polh−/− mice produced decreased serum titers of high‐affinity antibodies against a T‐dependent antigen, which was associated with a significant reduction in the number of plasma cells secreting high‐affinity antibodies. Analysis of the V region revealed that aa substitutions caused by A:T mutations were greatly reduced in Polh−/− mice. These results demonstrate that POLH is a limiting factor for A:T mutations and contributes to the efficient diversification of Ig genes and affinity maturation of antibodies.
Pulmonary stromal cells induce the generation of regulatory DC attenuating T‐cell‐mediated lung inflammation
Authors: Li Qian , Guo Zhenhong , Xu Xiongfei , Xia Sheng , Cao Xuetao
Published: September 2008

The tissue microenvironment may affect the development and function of immune cells such as DC. Whether and how the pulmonary stromal microenvironment can affect the development and function of lung DC need to be investigated. Regulatory DC (DCreg) can regulate T‐cell response. We wondered whether such regulatory DC exist in the lung and what is the effect of the pulmonary stromal microenvironment on the generation of DCreg. Here we demonstrate that murine pulmonary stromal cells can drive immature DC, which are regarded as being widely distributed in the lung, to proliferate and differentiate into a distinct subset of DCreg, which express high levels of CD11b but low levels of MHC class II (I‐A), CD11c, secrete high amounts of IL‐10, NO and prostaglandin E2 (PGE2) and suppress T‐cell proliferation. The natural counterpart of DCreg in the lung with similar phenotype and regulatory function has been identified. Pulmonary stroma‐derived TGF‐β is responsible for the differentiation of immature DC to DCreg, and DCreg‐derived PGE2 contributes to their suppression of T‐cell proliferation. Moreover, DCreg can induce the generation of CD4+CD25+Foxp3+ Treg. Importantly, infusion with DCreg attenuates T‐cell‐mediated eosinophilic airway inflammation in vivo. Therefore, the pulmonary microenvironment may drive the generation of DCreg, thus contributing to the maintenance of immune homoeostasis and the control of inflammation in the lung.
Reduction of cPLA2α overexpression: An efficient anti‐inflammatory therapy for collagen‐induced arthritis
Authors: Raichel Lior , Berger Slava , Hadad Nurit , Kachko Leonid , Karter Maria , Szaingurten‐Solodkin Irit , Williams Richard O. , Feldmann Marc , Levy Rachel
Published: September 2008

Cytosolic phospholipase A2α (cPLA2) plays an important role in the development of several inflammatory diseases. The aim of the present study is to determine whether inhibition of cPLA2 expression, using specific antisense oligonucleotides against cPLA2 (antisense), is efficient in reducing inflammation after its development. Two mouse models of inflammation were included in the study: thioglicolate peritonitis and collagen‐induced arthritis (CIA). The antisense was found to be specific and efficient in inhibiting cPLA2 expression and NADPH oxidase activity ex vivo in peritoneal phagocytes. Immunoblotting and immunohistochemistry analysis showed a significant elevation in cPLA2 expression in the inflamed joints of collagen‐induced arthritis mice localized in cell infiltrate, chondrocytes and the surrounding skin and skeletal muscle. Similarly, the cPLA2 metabolite, leukotriene B4, accumulated in the peritoneal cavity of mice with peritonitis. Inhibition of elevated cPLA2 expression after development of inflammation by intravenous administration of antisense resulted in a dramatic reduction in inflammation and a significant reduction in neutrophils recruitment to the site of inflammation in both mouse models of inflammation. Our results demonstrate the critical role of cPLA2 for the duration of inflammation and suggest that inhibition of cPLA2 expression by antisense oligonucleotides may serve as an efficient treatment of inflammatory diseases.
Seismic performance of a 3D full‐scale high‐ductile steel–concrete composite moment‐resisting frame—Part II: Test results and analytical validation
Authors: Braconi A. , Bursi O. S. , Fabbrocino G. , Salvatore W. , Taucer F. , Tremblay R.
Published: September 2008

This paper presents the results of a multi‐level pseudo‐dynamic seismic test program that was performed to assess the performance of a full‐scale three‐bay, two‐storey steel–concrete composite moment‐resisting frame built with partially encased composite columns and partial‐strength beam‐to‐column joints. The system was designed to develop a ductile response in the joint components of beam‐to‐column joints including flexural yielding of beam end plates and shear yielding of the column web panel zone. The ground motion producing the damageability limit state interstorey drift caused minor damage while the ultimate limit state ground motion level entailed column web panel yielding, connection yielding and plastic hinging at the column base connections. The earthquake level chosen to approach the collapse limit state induced more damage and was accompanied by further column web panel yielding, connection yielding and inelastic phenomena at column base connections without local buckling. During the final quasi‐static cyclic test with stepwise increasing displacement–amplitudes up to an interstorey drift angle of 4.6%, the behaviour was ductile although cracking of beam‐to‐end‐plate welds was observed. Correlations with numerical simulations taking into account the inelastic cyclic response of beam‐to‐column and column base joints are also presented in the paper together. Inelastic static pushover and time history analysis procedures are used to estimate the structural behaviour and overstrength factors of the structural system under study. Copyright © 2008 John Wiley & Sons, Ltd.
Vascular risk and depression in the Hispanic Established Population for the Epidemiologic Study of the Elderly (EPESE)
Authors: Zimmerman Jennifer A. , Mast Benjamin T. , Miles Toni , Markides Kyriakos S.
Published: September 2008

Objective

Although vascular depression has received considerable research attention, relatively little research in this area has focused on minority samples. This study investigated the association between baseline vascular risk factors (VRFs) and risk for elevated depressive symptoms at 2‐year follow‐up in a sample of 964 individuals without significant depressive symptomotology (CES‐D < 12) or cognitive impairment (MMSE≥ 24) at baseline from the Hispanic Established Population for the Epidemiologic Study of the Elderly.

Methods

We examined the associations between self‐reported baseline vascular risk factors (chest pain, heart attack, stroke, hypertension, diabetes, and smoking) and a composite of these risk factors with elevated depressive symptoms (CES‐D ≥ 16) at 2‐year follow‐up.

Results

Seventy‐four (7.7%) of the 964 participants without evidence of depression at baseline demonstrated elevated depressive symptoms (CESD ≥ 16) 2 years later. There was an overall pattern of higher rates of elevated depressive symptoms at 2‐year follow‐up with increasing number of vascular risk factors (0 VRFs = 6.4%, 1 VRF = 5.5%, 2 VRFs = 7.7%, and 3 or more VRFs = 14.7%). After controlling for demographic variables, physical functioning, and other medical conditions, the cumulative vascular risk index was significantly associated with elevated depressive symptoms at 2‐year follow‐up (p < 0.05).

Conclusions

Our results suggest vascular conditions may contribute to risk for depression over time among Mexican American elders, and this is relatively independent of other medical conditions. These findings suggest that depression is an additional long‐term complication of these common cardiovascular disorders. Copyright © 2008 John Wiley & Sons, Ltd.
Predictors of response to sertraline in patients with major depression
Authors: Morishita Shigeru , Kinoshita Toshihiko
Published: September 2008

Objective

Investigation of the characteristics of patients being treated with antidepressants would be useful in determining which patients would most likely benefit from antidepressant. The purpose of this study was to examine the possible predictors of response to sertraline in major depression.

Method

A retrospective cohort analysis was carried out among major depression patients. Eighty two patients were identified who were receiving sertraline to treat major depression.

Results

The cumulative percentage of responders was over 80% after 6 weeks, and the cumulative percentage of responders was over 80% in patients receiving a 75 mg daily dose. Clinical factors, including age, gender, frequency of episodes, and family history were examined as possible predictors of response to sertraline. On Cox proportional hazards analysis, age, and gender were independent predictive factors of improvement with sertraline. The most influential factor was age (ecoef = 1.894), followed by gender (ecoef = 0.542); age over 40 years (χ2 = 5.598, df = 1,p = 0.018) and female gender (χ2 = 7.370, df = 1,p = 0.0066) were significantly associated with improvement.

Conclusions

A 6‐week treatment period, a 75 mg daily dose, age over 40 years, and female gender appear to be predictors of response to sertraline in major depression. These factors should guide clinicians in determining the choice of antidepressant. Copyright © 2008 John Wiley & Sons, Ltd.
Hypomethylation of retrotransposable elements correlates with genomic instability in non‐small cell lung cancer
Authors: Daskalos Alexandros , Nikolaidis Georgios , Xinarianos George , Savvari Paraskevi , Cassidy Adrian , Zakopoulou Roubini , Kotsinas Athanasios , Gorgoulis Vassilis , Field John K. , Liloglou Triantafillos
Published: September 2008

LINE‐1 and Alu elements are non‐LTR retrotransposons, constituting together over 30% of the human genome and they are frequently hypomethylated in human tumors. A relationship between global hypomethylation and genomic instability has been shown, however, there is little evidence to suggest active role for hypomethylation‐mediated reactivation of retroelements in human cancer. In our study, we examined by Pyrosequencing the methylation levels of LINE‐1 and Alu sequences in 48 primary nonsmall cell carcinomas and their paired adjacent tissues. We demonstrate a significant reduction of the methylation levels of both elements (p = 7.7 × 10−14 and 9.6 × 10−7, respectively). The methylation indices of the 2 elements correlated (p = 0.006), suggesting a possible common mechanism for their methylation maintenance. Genomic instability was measured utilizing 11 fluorescent microsatellite markers located on lung cancer hot‐spot regions such as 3p, 5q 9p, 13q and 17p. Hypomethylation of both transposable elements was associated with increased genomic instability (LINE, p = 7.1 × 10−5; Alu, p = 0.008). The reduction of the methylation index of LINE‐1 and Alu following treatment of 3 lung cell lines with 5‐aza‐2′‐deoxycitidine, consistently resulted in increased expression of both elements. Our study demonstrates the strong link between hypomethylation of transposable elements with genomic instability in non‐small cell lung cancer and provides early evidence for a potential active role of these elements in lung neoplasia. As demethylating agents are now entering lung cancer trials, it is imperative to gain a greater insight into the potential reactivation of silent retrotransposons in order to advance for the clinical utilization of epigenetics in cancer therapy. © 2008 Wiley‐Liss, Inc.
How to minimize formation and growth of tumours: Potential benefits of decapod crustaceans for cancer research
Authors: Vogt Günter
Published: September 2008

Tumours have only rarely been observed in the decapod crustaceans, a large animal group of more than 10,000 species that includes the commercially important and well investigated shrimp, lobsters, crayfish and crabs. Analysis of the literature and information from cancer and diseases data bases revealed a total of 15 incidences, some of them being questionable. Even in the long‐lived species, which can reach life spans of almost 100 years, neoplasias are virtually unknown. The data published so far suggest that the strikingly different frequencies of carcinogenesis between decapods and other well investigated animal groups like mammals, fish, insects and molluscs is based on differences of the metabolic pathways for carcinogens, the immune systems, and the regulation of stem cells. Therefore, representatives of the Decapoda may serve as useful models to study how organisms can successfully prevent or control spontaneously and environmentally induced cell proliferation. A particularly promising candidate for in‐depth investigation of these topics is the marbled crayfish, a rather new clonal lineage that is presently being introduced as a laboratory model in development and epigenetics. © 2008 Wiley‐Liss, Inc.
Dietary fats altered nephrotoxicity profile of methylmercury in rats
Authors: Jin Xiaolei , Lok Eric , Caldwell Don , Mueller Rudi , Kapal Kamla , Liston Virginia , Kubow Stan , Chan Hing Man , Mehta Rekha
Published: September 2008

Weanling male Sprague–Dawley rats were administered semi‐purified isocaloric diet containing soy oil (SO), seal oil (SE), docosahexaenoic acid (DHA), fish oil (FO) or lard (LA) for 28 days, and then gavaged with 0, 1 or 3 mg MeHg kg−1body weight per day and fed the same diet for 14 days. Serum and 24 h urine samples were collected on the day of necropsy, and analyzed for markers of kidney function and diseases. Kidney slices were analyzed for para‐amino‐hippurate (PAH) and tetraethylammonium (TEA) uptake, total mercury and MeHg content, and examined for pathological lesions. Total mercury and MeHg contents increased significantly and dose‐dependently in all dietary groups. MeHg significantly increased relative kidney weight in all groups, serum creatinine in all except SO group, serum uric acid in the DHA and LA groups, serum Mg in all except the LA group, and urinary protein in the SO group. MeHg significantly decreased serum urea nitrogen in SE, FO and LA groups, urinary creatinine in the DHA group, PAH uptake in all except the SE group, and TEA uptake in all groups. MeHg caused nephrosis in all dietary groups. MeHg also significantly increased neutrophil counts in all except the SE group, decreased serum albumin and triglyceride in all except the DHA group, and increased serum total cholesterol in all groups, suggesting a nephrotic syndrome‐like outcome. These results confirmed that kidney tubules are major targets of MeHg nephrotoxicity. Treatment with dietary fats did not prevent, but rather altered the profile of, nephrotoxicity of MeHg in rats. Copyright © 2008 John Wiley & Sons, Ltd.