bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of a Proton Conducting Filler on the Physico‐Chemical Properties of SPEEK‐Based Membranes
Authors: Mecheri B. , D'Epifanio A. , Pisani L. , Chen F. , Traversa E. , Weise F. C. , Greenbaum S. , Licoccia S.
Published: August 2009

Composite membranes based on sulphonated polyetherether ketone (SPEEK) having a 60% degree of sulphonation (DS = 0.6) and containing 23 and 50 wt.‐% hydrated tin oxide (SnO2·nH2O) were prepared and characterised. The lower water uptake (WU) and the higher conductivity values recorded for the composite membranes with respect to pure SPEEK reference suggested the involvement of SnO2·nH2O in the proton conduction mechanism. Pulsed‐field‐gradient spin‐echo (PFGSE) NMR was employed to obtain a direct measurement of water self‐diffusion coefficient in the membranes. Differences were observed between the unfilled SPEEK and the composites, including departures from the normal correlation between water diffusivity and proton conductivity in the case of composites. To better understand the SnO2·nH2O effect on the proton transport properties of the SPEEK‐based membrane, we employed an analytical model that predicts the membrane conductivity as a function of its hydration level and porous structure. The comparison of the model results with the experimental proton conductivity values demonstrated that the tin oxide phase provides additional paths between the water clusters for proton transport, resulting in reduced tortuosity and enhanced proton conductivity. Moreover, the composite showed reduced methanol crossover with respect to the unfilled membrane.
The determinants of health‐care expenditure: new results from semiparametric estimation
Authors: Herwartz Helmut , Theilen Bernd
Published: August 2009

Recent cross‐country studies have questioned the existence of a systematic relationship between per capita health‐care expenditure (HCE) and explanatory variables such as income, population ageing and total public expenditure. We reexamine this issue mainly focussing at a flexible semiparametric estimation method that allows the parameters of the model to depend on a state variable. Using the age structure of the population as the state variable, we find that the income elasticity increases with population ageing, while other explanatory variables are not significantly influenced by it. Additionally we find that the HCE relationship becomes more and more unstable in ageing economies. These results explain the difficulty to identify both the influence of population ageing and income on HCE in previous studies. Furthermore they indicate that international convergence of HCE across countries crucially depends on the convergence of the population age structure. We also discuss the policy implications of our results regarding the design of a fair health‐care financing system and the evolution of HCE to avoid budgetary problems. Copyright © 2009 John Wiley & Sons, Ltd.
Comments on contingency management and conditional cash transfers
Authors: Higgins Stephen T.
Published: August 2009

This essay discusses research on incentive‐based interventions to promote healthy behavior change, contingency management (CM) and conditional cash transfers (CCT). The overarching point of the essay is that CM and CCT are often treated as distinct areas of inquiry when at their core they represent a common approach. Some potential bi‐directional benefits of recognizing this commonality are discussed. Distinct intellectual traditions probably account for the separate paths of CM and CCT to date, with the former being rooted in behavioral psychology and the latter in microeconomics. It is concluded that the emerging field of behavioral economics, which is informed by and integrates principles of each of those disciplines, may provide the proper conceptual framework for integrating CM and CCT. Copyright © 2009 John Wiley & Sons, Ltd.
Transient storage processes in a steep headwater stream
Authors: Scordo Elisa B. , Moore R. Dan
Published: August 2009

This study examined hyporheic exchange flow and transient storage processes within a steep headwater stream, using both hydrometric and tracer approaches. Vertical hydraulic gradients varied systematically with location within step‐pool units, with upwelling located in a relatively narrow zone immediately below the step. Downward hydraulic gradients were greatest immediately above channel steps, and in that zone, gradients were also correlated with step height. Hydraulic conductivity tended to be lower in downwelling zones than in upwelling and neutral locations. Transient storage areas determined from reach‐scale tracer tests tended to increase with discharge, whereas transient exchange coefficients were not correlated with discharge. Transient storage exchange fluxes estimated from the reach‐scale tracer tests were an order of magnitude greater than hyporheic exchange fluxes estimated by scaling up local estimates based on Darcy's Law and piezometer measurements. This difference could be explained by the presence of lateral hyporheic flow paths and/or the influences of transient storage in pools. Tracer injections into infiltrometers using Rhodamine WT and sodium chloride allowed visual determination of hyporheic discharge zones and calculation of residence times for hyporheic flow through the step (pool inflow) and transport through the downstream pool (pool outflow). These tests indicated that residence time in pools can be comparable to hyporheic residence times, although the lack of replication in time and space limits the drawing of strong inferences. Copyright © 2009 John Wiley & Sons, Ltd.
The impact of Quaternary sea level changes on the evolution of the Amazonian lowland
Authors: Irion Georg , Müller Jens , Morais Jáder O. , Keim Gertrud , de Mello Jose Nunes , Junk Wolfgang J.
Published: August 2009

Coring and acoustic surveying (3·5 kHz) in both rivers and lakes in Central Amazonia provide additional insights into the Late Quaternary hydrological and sedimentological development of the Amazon River and its tributaries. Erosion and accumulation phases were found to be linked to Quaternary sea level changes. The low sea level phase during the last glacial maximum caused deep incision of the Amazon River and erosion in major tributaries such as the Rio Negro, 1500 km upstream from the Amazon mouth. A 3·5‐kHz profiling suggests a lowering of the water level by at least 30 m at Manaus. During that phase, the slope of the Amazon valley must have increased, resulting in an increase in bed load transport capacity. The subsequent sea level rise caused a backwater effect far upstream, with silting up of the Amazon valley and the tributary inflows. Former river systems changed into Ria lakes. The floodplains of the Amazon River, the várzea, were formed approximately 5000 years ago when the sea level approached its present‐day level. Copyright © 2009 John Wiley & Sons, Ltd.
Effect of atmospheric pressure and temperature on entrapped gas content in peat
Authors: Waddington J. M. , Harrison K. , Kellner E. , Baird A. J.
Published: August 2009

Entrapped biogenic gas in peat can greatly affect peatland biogeochemical and hydrological processes by altering volumetric water content, peat buoyancy, and ‘saturated’ hydraulic conductivity, and by generating over‐pressure zones. These over‐pressure zones further affect hydraulic gradients which influence water and nutrient flow direction and rate. The dynamics of entrapped gas are of global interest because the loss of this gas to the atmosphere via ebullition (bubbling) is likely the dominant transport mechanism of methane (CH4) to the atmosphere from peatlands, which are the largest natural terrestrial source per annum of atmospheric CH4. We investigated the relationship between atmospheric pressure and temperature on volumetric gas content (VGC) and CH4 ebullition using a laboratory peat core incubation experiment.

Peat cores were incubated at three temperatures (one core at 4 °C, three cores at 11 °C, and one core at 20 °C) in sealed PVC cylinders, instrumented to measure VGC, pore‐water CH4 concentrations, and ebullition (volume and CH4 concentrations). Ebullition events primarily occurred (71% of the time) during periods of falling atmospheric pressure. The duration of the drop in atmospheric pressure had a larger control on ebullition volume than the magnitude of the drop. VGC in the 20 °C core increased from the onset of the experiment and reached a fluctuating but time‐averaged constant level between experiment day 30 and 115. The change in VGC was low for the 11 °C cores for the initial period of the experiment but showed large increases when the growth chamber temperature increased to 20 °C due to a malfunction. The core maintained at 4 °C showed only a small increase in entrapped gas content throughout the experiment. The 20 °C core showed the largest increase in VGC. The increases in VGC occurred despite pore‐water concentrations of CH4 being below the equilibrium solubility level. Copyright © 2009 John Wiley & Sons, Ltd.
Intracellular imaging of host‐pathogen interactions using combined CARS and two‐photon fluorescence microscopies
Authors: Robinson Iain , Ochsenkühn Michael Andreas , Campbell Colin J. , Giraud Gerard , Hossack William J. , Arlt Jochen , Crain Jason
Published: August 2009

Intracellular imaging is a key tool in the investigation of host‐pathogen interactions. Advances in this area are particularly sought to understand the effect of viral infection processes on the host cell and its metabolic functions including those cases where host cell lipid metabolism is modulated as a result of infection. We demonstrate the use of combined coherent anti‐Stokes Raman scattering (CARS) and two‐photon fluorescence microscopies to image fibroblast cells infected by cytomegalovirus. CARS is used to image the host cell membrane, lipid droplets and morphology of the nucleus. Cell nuclei are found to expand during infection, approximately doubling in area. Some cells also show accumulations of lipid droplets at the nuclear periphery. Using a genetically modified virus strain expressing the green fluorescent protein also enables two‐photon imaging of the same cells to reveal the location, nature and extent of viral protein expression. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
A new strategy to improve the predictive ability of the local lazy regression and its application to the QSAR study of melanin‐concentrating hormone receptor 1 antagonists
Authors: Li Jiazhong , Li Shuyan , Lei Beilei , Liu Huanxiang , Yao Xiaojun , Liu Mancang , Gramatica Paola
Published: August 2009

In the quantitative structure‐activity relationship (QSAR) study, local lazy regression (LLR) can predict the activity of a query molecule by using the information of its local neighborhood without need to produce QSAR models a priori. When a prediction is required for a query compound, a set of local models including different number of nearest neighbors are identified. The leave‐one‐out cross‐validation (LOO‐CV) procedure is usually used to assess the prediction ability of each model, and the model giving the lowest LOO‐CV error or highest LOO‐CV correlation coefficient is chosen as the best model. However, it has been proved that the good statistical value from LOO cross‐validation appears to be the necessary, but not the sufficient condition for the model to have a high predictive power. In this work, a new strategy is proposed to improve the predictive ability of LLR models and to access the accuracy of a query prediction. The bandwidth of k neighbor value for LLR is optimized by considering the predictive ability of local models using an external validation set. This approach was applied to the QSAR study of a series of thienopyrimidinone antagonists of melanin‐concentrating hormone receptor 1. The obtained results from the new strategy shows evident improvement compared with the commonly used LOO‐CV LLR methods and the traditional global linear model. © 2009 Wiley Periodicals, Inc. J Comput Chem, 2010
Purification of adenoviral vectors by combined anion exchange and gel filtration chromatography
Authors: Eglon Marc N. , Duffy Aoife M. , O'Brien Timothy , Strappe Padraig M.
Published: August 2009

Background

Adenoviral vectors are used extensively in human gene therapy trials and in vaccine development. Large‐scale GMP production requires a downstream purification process, and liquid chromatography is emerging as the most powerful mode of purification, enabling the production of vectors at a clinically relevant scale and quality. The present study describes the development of a two‐step high‐performance liquid chromatography (HPLC) process combining anion exchange (AIEX) and gel filtration (GF) in comparison with the caesium chloride density gradient method.

Methods

HEK‐293 cells were cultured in ten‐layer CellStacks™ and infected with 10 pfu/cell of adenoviral vector expressing green fluorescent protein (Ad5‐GFP). Cell‐bound virus was harvested and benzonase added to digest DNA, crude lysate was clarified by centrifugation and filtration prior to HPLC. Chromatography fractions were added to HEK‐293 cells and GFP expression measured using a fluorescent plate reader.

Results

Using AIEX then GF resulted in an adenoviral vector with purity comparable to Ad5‐GFP purified by CsCl, whereas the reverse process (GF–AIEX) showed a reduced purity by electrophoresis and required further buffer exchange of the product. The optimal process (AIEX–GF) resulted in a vector yield of 2.3 × 107 pfu/cm2 of cell culture harvested compared to 3.3 × 107 pfu/cm2 for CsCl. The process recovery for the HPLC process was 36% compared to 27.5% for CsCl and total virion to infectious particle ratios of 18 and 11, respectively, were measured.

Conclusions

We present a simple two‐step chromatography process that is capable of producing high‐quality adenovirus at a titre suitable for scale‐up and clinical translation. Copyright © 2009 John Wiley & Sons, Ltd.
AFM‐ and NSOM‐based force spectroscopy and distribution analysis of CD69 molecules on human CD4+ T cell membrane
Authors: Hu Mingqian , Chen Jianan , Wang Jiongkun , Wang Xiaoping , Ma Shuyuan , Cai Jiye , Chen Crystal Y. , Chen Zheng W.
Published: August 2009

Although CD69 is well known as an early T cell‐activation marker, the possibility that CD69 are distributed as nano‐structures on membrane for immune regulation during T cell activation has not been tested. In this study, nanoscale features of CD69 expression on activated T cells were determined using the atomic force microscopy (AFM) topographic and force‐binding nanotechnology as well as near‐field scanning optical microscopy (NSOM)‐/fluorescence quantum dot (QD)‐based nanosacle imaging. Unstimulated CD4+ T cells showed neglectable numbers of membrane CD69 spots binding to the CD69 Ab‐functinalized AFM tip, and no detectable QD‐bound CD69 as examined by NSOM/QD‐based imaging. In contrast, Phytohemagglutinin (PHA)‐activated CD4+ T cells expressed CD69, and displayed many force‐binding spots binding to the CD69 Ab‐functionalized AFM tip on about 45% of cell membrane, with mean binding‐rupture forces 276 ± 71 pN. Most CD69 molecules appeared to be expressed as 100–200 nm nanoclusters on the membrane of PHA‐activated CD4+ T cells. Meanwhile, NSOM/QD‐based nanoscale imaging showed that CD69 were non‐uniformly distributed as 80–200 nm nanoclusters on cell‐membrane of PHA‐activated CD4+ T cells. This study represents the first demonstration of the nano‐biology of CD69 expression during T cell activation. Copyright © 2009 John Wiley & Sons, Ltd.
Raman and IR spectra of the cation‐deficient Aurivillius layered crystal Bi2W2O9
Authors: Ma̧czka M. , Macalik L. , Hanuza J.
Published: August 2009

Raman and IR spectra of Bi2W2O9, the m = 2 member of the cation‐deficient Aurivillius phase, have been measured. The obtained spectra are discussed using the factor group approach for the orthorhombic Pna21(C2v9) space group. The results are compared with those obtained for Bi2WO6 and some m = 2 members of Aurivillius family of bismuth layered ferroelectrics. Copyright © 2009 John Wiley & Sons, Ltd.
Preparation and characterization of sizable macroporous epoxy resin‐based monolithic supports for flow‐through systems
Authors: Nguyen Anh Mai , Dinh Ngoc Phuoc , Cam Quach Minh , Sparrman Tobias , Irgum Knut
Published: August 2009

This paper presents further results from our efforts to prepare sizable macroporous monolithic materials from epoxy resins and polyamines by emulsion polymerization. For their uses as supports in flow systems, the study aimed at developing materials possessing maximum fluid permeability, high mechanical stability, and a controlled porosity and surface area. Characterization of the materials has been carried out using different techniques, focusing on morphological and mechanical features, and on the surface chemistry. Morphology and porosity were studied with SEM, nitrogen adsorption/desorption, mercury intrusion porosimetry (MIP), and 2H NMR cryoporosimetry. The chemical composition of the bulk structures and their surfaces was studied by means of bulk elemental analysis and X‐ray photoelectron spectroscopy, and potentiometric titration was used to assess the relative amounts of amines and epoxy groups. Essentially, the morphological features were a high fluid permeability, but rather low specific surface area. Convective flow was facilitated by large, interconnected, and evenly spaced macropores which were formed by nonporous skeletons of the connected‐rod type. Despite the interfacial nature of the polymerization, the bulk and the surface of the fully cured materials showed similar elemental compositions. All materials were found to have a high surface density of hydroxyl groups, which facilitates functionalization reactions.
Functionalization of epoxy‐based monoliths for ion exchange chromatography of proteins
Authors: Dinh Ngoc Phuoc , Cam Quach Minh , Nguyen Anh Mai , Shchukarev Andrei , Irgum Knut
Published: August 2009

Macroporous epoxy‐based monoliths prepared by emulsion polymerization have been modified for use in ion exchange chromatography (IEC) of proteins. Strong anion exchange functionality was established by iodomethane quaternization of tertiary amine present on the monolith surface as a part of the polymer backbone. The modification pathway to cation exchange materials was via incorporation of glycidyl methacrylate (GMA) brushes which were coated using atom transfer radical polymerization (ATRP). Strong (SO3–) and weak (COO–) cation exchange groups were thereafter introduced onto the GMA‐grafted monoliths by reactions with sodium hydrogen sulfite and iminodiacetic acid, respectively. Grafting was confirmed by XPS, gravimetric measurement, and chromatographic behavior of the modified materials toward model proteins. In incubation experiments the proteins were recovered quantitatively with no obvious signs of unfolding after contact with the stationary phase for >2 h. Chromatographic assessments on the functionalized columns as well as problems associated with flow‐through modification by ATRP are discussed.
Methodologies for soil erosion and land degradation assessment in mediterranean‐type ecosystems
Authors: Ries J. B.
Published: August 2009

The detection of spatial distribution and temporal development of soil erosion processes and forms on abandoned land still represents a great scientific challenge. Abandoned fields as a result of land use changes during the past century can frequently be found in the Mediterranean. The former fields are generally used for grazing and therefore are highly complex concerning the variability of soil erosion processes.

A three‐step research method is used for assessment and evaluation of soil erosion processes. By means of rainfall simulations, test plot monitoring and large‐scale aerial photography, erosion processes are quantified and scenarios are presented for three test sites in northeast Spain.

With a large‐scale test plot monitoring the development of soil erosion processes can be evaluated. Areas with predominantly increasing soil erosion activity are classified as unstable, whereas areas with unchanged or decreasing soil erosion activity are classified as vulnerable or stable. Forty to 70 years after abandonment and change to grazing all observed test areas show a progressive degradation. The proportion of unstable areas reaches around 50 per cent. The extension of unstable areas, characterised by a combination of increasing erosion activity and decreasing vegetation cover, is highest within the semi‐arid Ebro Basin reaching 29 per cent of the whole area. Dense vegetation cover does not always correlate with stability in erosion and increase in cover does not generally lead to a stabilisation of erosion processes. Even under increasing vegetation cover of more than 60 per cent high erosion and runoff rates are possible particularly in connected rills and on sheep trails. Therefore, grazing on abandoned fields is the critical factor leading to further degradation. Copyright © 2009 John Wiley & Sons, Ltd.
Venous drainage in retrograde island flap: An experimental study using fluorescence tracing technique
Authors: Jing Zhi‐Zhen , Chang Shi‐Min , You Mu‐Rong , Yu Guang‐Rong
Published: August 2009

The pathway of venous drainage in retrograde island flaps was investigated by fluorescence tracing technique using the saphenous fasciocutaneous flap in New Zealand White rabbits. Forty animals were allocated into four groups according to the different times at 30 minutes (I), 24 hours (II), 72 hours (III), and 7 days (IV) after the operation. According to the different routes to give tracer, each group was further allocated into two subgroups of the artery injection and vein injection. For each animal, one hindlimb was assigned as the experimental side, the contralateral side as control without giving tracer. The erythrocytes were separated, labeled with fluorescein isothiocyanate (FITC), detected, and injected into the artery or vein. Subsequently, the flaps were harvested 5 seconds after injection and immediately frozen, sectioned, and observed under microscope. In group I and II, the fluorescence was observed mainly around the vessel adventitia of the vein and artery and tunica intima of the artery. In group III, there was weak fluorescence observed in the lumen of vein. In group IV, fluorescence was distributed principally in the lumen of the vein. In addition, fluorescence was not observed in the saphenous nerve in group I and there was mild fluorescence in the saphenous nerve in groups II, III, and IV. These findings suggest that the venous return is through “bypass route” in earlier period. In later period, the venous retrograde return is through “bypass route” and “incompetent valves route;” however, “incompetent valves route” becomes the main route. © 2009 Wiley‐Liss, Inc. Microsurgery 2010.
Temporomandibular joint, skull base and mandibular ramus functional reconstruction with homologous bank tissue and free flap: A case report with 30 months follow‐up
Authors: Maranzano Massimo , Rizzo Roberto , Cicognini Alessandro , Sorato Renzo , Recchia Guglielmo , De Grazia Raffaella , Di Paola Francesco , Paolin Adolfo , Mazzoleni Giovanni
Published: August 2009

Big craniofacial resections for highly invasive malignant neoplasm, including skull base and maxillary bones, always represent a difficult chance for the reconstructive surgeon. In these cases it is not easy to restore anatomy and function simultaneously even adopting complex microsurgical techniques. In maxillofacial and oral surgery, simple bone homotransplantation for small bone segments reconstruction has been developing as popular technique and tissue banks offer not only bone segments but also many different tissues including complex body parts. In this paper we present, a case report of a homotransplantation of a complete temporomandibular joint (TMJ) together with a portion of the medial skull base and mandibular ramus folded with an ante‐brachial fascio‐periosteal free flap as secondary reconstruction after nearly 5 years from the removal of a sarcoma of the TMJ involving the skull base and a follow up of more than 30 months. © 2009 Wiley‐Liss, Inc. Microsurgery 2010.
Integrated network experimentation for QoS measurements in opaque MANETs
Authors: Biswas Pratik K. , Poylisher Alex , Chadha Ritu , Ghosh Abhrajit
Published: August 2009

Integrated network experimentation often combines real nodes with simulated ones, each modeling different portions of the topology, in the same experimental run. They enable new validation techniques and larger experiments than obtainable using real elements alone. Integrated experiments can be particularly useful in testing and validating QoS mechanisms for mobile ad hoc networks (MANETs), when the characteristics of the intermediate wireless network segments are not observable from the end segments, and as a consequence these intermediate segments have to be treated as opaque networks; and also when there is a need to conduct experiments in a real MANET environment at a scale larger than just a few nodes. End‐to‐end QoS assurance for such opaque networks, consisting of admission control and quality adjustment, can be based on techniques for dynamically measuring throughput representing the state of these networks. In this paper, we describe a distributed and hybrid testbed that has been deployed for running large‐scale integrated experiments to demonstrate the efficacy of a measurement‐based QoS solution. The infrastructure for the testbed provides an integrated platform consisting of real nodes running the actual software under test, augmented with a simulated network environment. We define a set of metrics and run experiments to evaluate the effectiveness of the QoS solution as well the performance of the deployed testbed. We propose an alternative architecture that employs a Xen‐based virtualization of the real nodes from the deployed testbed. We compare the performances of the virtualized architecture with the deployed architecturevis‐à‐vislatency and resource utilization. Our goal is to establish benchmarks for running large‐scale experiments on performance and QoS measurements in virtualized environments. Copyright © 2009 John Wiley & Sons, Ltd.
Analytical solutions to nonlinear equations arising in heat transfer by variational iteration, homotopy perturbation, and Adomian decomposition methods
Authors: Ganji D.D. , Gorji M. , Alipanah M. , Farnad E.
Published: August 2009

As thermal conductivity plays an important role on fin efficiency, we tried to solve heat transfer equation with thermal conductivity as a function of temperature. In this research, some new analytical methods called homotopy perturbation method, variational iteration method, and Adomian decomposition method are introduced to be applied to solve the nonlinear heat transfer equations, and also the comparison of the applied methods (together) is shown graphically. © 2009 Wiley Periodicals, Inc. Numer Methods Partial Differential Eq 2010
Numerical algorithms for solutions of Korteweg–de Vries equation
Authors: Korkmaz Alper
Published: August 2009

The nonlinear Korteweg–de Vries (KdVE) equation is solved numerically using both Lagrange polynomials based differential quadrature and cosine expansion‐based differential quadrature methods. The first test example is travelling single solitary wave solution of KdVE and the second test example is interaction of two solitary waves, whereas the other three examples are wave production from solitary waves. Maximum error norm and root mean square error norm are computed, and numerical comparison with some earlier works is done for the first two examples, the lowest four conserved quantities are computed for all test examples. © 2009 Wiley Periodicals, Inc. Numer Methods Partial Differential Eq 2010
Synthesis of sub‐micrometer core–shell rubber particles with 1,2‐azobisisobutyronitrile as initiator and deformation mechanisms of modified polystyrene under various conditions
Authors: Dai Rujun , Gao Guanghui , Sun Shulin , Tan Zhiyong , Zhang Huixuan
Published: August 2009

BACKGROUND: Sub‐micrometer core‐shell polybutadiene‐graft‐polystyrene (PB‐g‐PS) copolymers with various ratios of polybutadiene (PB) core to polystyrene (PS) shell were synthesized by emulsion grafting polymerization with 1,2‐azobisisobutyronitrile (AIBN) as initiator. These graft copolymers were blended with PS to prepare PS/PB‐g‐PS with a rubber content of 20 wt%. The mechanical properties, morphologies of the core‐shell rubber particles and deformation mechanisms under various conditions were investigated.

RESULTS: Infrared spectroscopic analysis confirmed that PS could be grafted onto the PB rubber particles. The experimental results showed that a specimen with a ‘cluster’ dispersion state of rubber particles in the PS matrix displayed better mechanical properties. Transmission electron micrographs suggested that crazing only occurred from rubber particles and extended in a bridge‐like manner to neighboring rubber particles parallel to the equatorial plane at a high speed for failure specimens, while the interaction between crazing and shear yielding stabilized the growing crazes at a low speed in tensile tests.

CONCLUSION: AIBN can be used as an initiator in the graft polymerization of styrene onto PB. The dispersion of rubber particles in a ‘cluster’ state leads to better impact resistance. The deformation mechanism in impact tests was multi‐crazing, and crazing and shear yielding absorbed the energy in tensile experiments. Copyright © 2009 Society of Chemical Industry
Improving 2‐DE gel image denoising using contourlets
Authors: Tsakanikas Panagiotis , Manolakos Elias S.
Published: August 2009

One of the most commonly used methods for protein separation is 2‐DE. After 2‐DE gel scanning, images with a plethora of spot features emerge that are usually contaminated by inherent noise. The objective of the denoising process is to remove noise to the extent that the true spots are recovered correctly and accuratelyi.e. without introducing distortions leading to the detection of false‐spot features. In this paper we propose and justify the use of the contourlet transform as a tool for 2‐DE gel images denoising. We compare its effectiveness with state‐of‐the‐art methods such as wavelets‐based multiresolution image analysis and spatial filtering. We show that contourlets not only achieve better averageS/Nperformance than wavelets and spatial filters, but also preserve better spot boundaries and faint spots and alter less the intensities of informative spot features, leading to more accurate spot volume estimation and more reliable spot detection, operations that are essential to differential expression proteomics for biomarkers discovery.
Proposal for a standard system for drawing structural diagrams of N‐ and O‐linked carbohydrates and related compounds
Authors: Harvey David J. , Merry Anthony H. , Royle Louise , P. Campbell Matthew , Dwek Raymond A. , Rudd Pauline M.
Published: August 2009

Symbolic diagrams are commonly used to depict N‐ and O‐linked glycans but there is no general consensus as to how individual constituent monosaccharides or linkages are shown. This article proposes a system that avoids ambiguities inherent in most other systems and is appropriate for both hand drawing and computer applications. Constituent monosaccharides are depicted by shapes modified to show OAc, deoxy, etc. Linkage is indicated by the bond angle and anomericity by solid (β) or dashed (α) lines.
Proteomic analysis of cells in the early stages of herpes simplex virus type‐1 infection reveals widespread changes in the host cell proteome
Authors: Antrobus Robin , Grant Kyle , Gangadharan Bevin , Chittenden David , Everett Roger D. , Zitzmann Nicole , Boutell Chris
Published: August 2009

During infection by herpes simplex virus type‐1 (HSV‐1) the host cell undergoes widespread changes in gene expression and morphology in response to viral replication and release. However, relatively little is known about the specific proteome changes that occur during the early stages of HSV‐1 replication prior to the global damaging effects of virion maturation and egress. To investigate pathways that may be activated or utilised during the early stages of HSV‐1 replication, 2‐DE and LC‐MS/MS were used to identify cellular proteome changes at 6 h post infection. Comparative analysis of multiple gels representing whole cell extracts from mock‐ and HSV‐1‐infected HEp‐2 cells revealed a total of 103 protein spot changes. Of these, 63 were up‐regulated and 40 down‐regulated in response to infection. Changes in selected candidate proteins were verified by Western blot analysis and their respective cellular localisations analysed by confocal microscopy. We have identified differential regulation and modification of proteins with key roles in diverse cellular pathways, including DNA replication, chromatin remodelling, mRNA stability and the ER stress response. This work represents the first global comparative analysis of HSV‐1 infected cells and provides an important insight into host cell proteome changes during the early stages of HSV‐1 infection.
Carminic acid: an antioxidant to protect erythrocytes and DNA against radical‐induced oxidation
Authors: Li Guo‐Xiang , Liu Zai‐Qun , Wu Di
Published: August 2009

This work explored the antioxidant effect of carminic acid (CarOH) on the oxidation of DNA and erythrocytes induced by 2,2′‐azobis(2‐amidinopropane hydrochloride) (AAPH). The half concentrations (IC50) of CarOH to scavenge radicals were measured by reacting with 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonate) radical cation (ABTS+•) and 2,2′‐diphenyl‐1‐picrylhydrazyl (DPPH). The values of IC50 were 8.0 and 26.0 µM when CarOH reacted with ABTS+• and DPPH, respectively. CarOH was able to protect DNA against AAPH‐induced oxidative damage by decreasing the formation rate of thiobarbituric acid reactive substance (TBARS). In addition, CarOH protected human erythrocytes against AAPH‐induced hemolysis concentration dependently, during which one molecule of CarOH can trap almost three radicals. Moreover, quantum calculation revealed that the hydroxyl group at 6‐position played major role in trapping radicals. Copyright © 2009 John Wiley & Sons, Ltd.
Alteration of oligomeric state and domain architecture is essential for functional transformation between transferase and hydrolase with the same scaffold
Authors: Koike Ryotaro , Kidera Akinori , Ota Motonori
Published: August 2009

Transferases and hydrolases catalyze different chemical reactions and express different dynamic responses upon ligand binding. To insulate the ligand molecule from the surrounding water, transferases bury it inside the protein by closing the cleft, while hydrolases undergo a small conformational change and leave the ligand molecule exposed to the solvent. Despite these distinct ligand‐binding modes, some transferases and hydrolases are homologous. To clarify how such different catalytic modes are possible with the same scaffold, we examined the solvent accessibility of ligand molecules for 15 SCOP superfamilies, each containing both transferase and hydrolase catalytic domains. In contrast to hydrolases, we found that nine superfamilies of transferases use two major strategies, oligomerization and domain fusion, to insulate the ligand molecules. The subunits and domains that were recruited by the transferases often act as a cover for the ligand molecule. The other strategies adopted by transferases to insulate the ligand molecule are the relocation of catalytic sites, the rearrangement of secondary structure elements, and the insertion of peripheral regions. These findings provide insights into how proteins have evolved and acquired distinct functions with a limited number of scaffolds.
The lily‐of‐the‐valley fragrance receptor—potential in prostate cancer imaging
Authors: Sturzu Alexander , Echner Hartmut , Heckl Stefan
Published: August 2009

BACKGROUND

The early diagnosis of prostate cancer and its metastases still remains a great challenge. Recently, olfactory receptors have been found in healthy and malignant prostate tissue. If conjugated to a dye, olfactory receptor ligands would represent candidates for markers of not only olfactory cells but also prostate tissue. Such a conjugate would be of great value for the detection of prostate cancer metastases.

METHODS

We coupled sulforhodamine, a fluorescence dye, to undecylic aldehyde, the antagonist of the lily‐of‐the‐valley fragrance receptor. By using confocal laser scanning microscopy and flow cytometry we examined the uptake of this conjugate into various different human cell lines. One healthy prostate cell line, two different prostate carcinoma cell lines, and five other carcinoma cell lines were investigated. CD1 nude mice bearing human PC3 prostate carcinoma xenografts were injected with the conjugate. Rhodamine fluorescence of mouse organ frozen sections was evaluated by confocal laser scanning microscopy.

RESULTS

When comparing the seven human cell lines, the conjugate was preferentially taken up by the cytoplasm of healthy and malignant prostate cells. The mice showed high conjugate uptake into the xenografts, but much lower uptake into the mouse organs. After 3 hr of circulation, efflux could be observed in the xenograft sections. Xenograft touch prints confirmed in vivo intracellular accumulation.

CONCLUSION

This conjugate may be of potential value in the diagnosis of prostate cancer and its metastases. Prostate 69: 1599–1602, 2009. © 2009 Wiley‐Liss, Inc.
Genistein and resveratrol, alone and in combination, suppress prostate cancer in SV‐40 tag rats
Authors: Harper Curt E. , Cook Leah M. , Patel Brijesh B. , Wang Jun , Eltoum Isam A. , Arabshahi Ali , Shirai Tomoyuki , Lamartiniere Coral A.
Published: August 2009

BACKGROUND

Chemoprevention utilizing dietary agents is an effective means to slow the development of prostate cancer. We evaluated the potential additive and synergistic effects of genistein and resveratrol for suppressing prostate cancer in the Simian Virus‐40 T‐antigen (SV‐40 Tag) targeted probasin promoter rat model, a transgenic model of spontaneously developing prostate cancer.

METHODS

Rats were fed genistein or resveratrol (250 mg/kg AIN‐76A diet) alone and in combination, and a low‐dose combination (83 mg genistein + 83 mg resveratrol/kg diet). Histopathology and mechanisms of action studies were conducted at 30 and 12 weeks of age, respectively.

RESULTS

Genistein, resveratrol, and the high‐dose combination treatments suppressed prostate cancer. The low‐dose combination did not elicit protection against prostate cancer and was most likely below the effective dose for causing significant histopathological changes. Total genistein and resveratrol concentrations in the blood reached 2,160 and 211 nM, respectively in rats exposed to the single treatments. Polyphenol treatments decreased cell proliferation and insulin‐like growth factor‐1 (IGF‐1) protein expression in the prostate. In addition, genistein as a single agent induced apoptosis and decreased steroid receptor coactivator‐3 (SRC‐3) in the ventral prostate (VP).

CONCLUSIONS

Genistein and resveratrol, alone and in combination, suppress prostate cancer development in the SV‐40 Tag model. Regulation of SRC‐3 and growth factor signaling proteins are consistent with these nutritional polyphenols reducing cell proliferation and increasing apoptosis in the prostate. Prostate 69: 1668–1682, 2009. © 2009 Wiley‐Liss, Inc.
Human prostate‐infiltrating CD8+ T lymphocytes are oligoclonal and PD‐1+
Authors: Sfanos Karen S. , Bruno Tullia C. , Meeker Alan K. , De Marzo Angelo M. , Isaacs William B. , Drake Charles G.
Published: August 2009

BACKGROUND

Prostate‐infiltrating CD8+ T lymphocytes (CD8+ PIL) are prevalent in men with prostate cancer (PCa), however, it is unclear whether the presence of such cells reflects a non‐specific immune infiltrate or an oligoclonal, antigen‐driven adaptive immune response.

METHODS

We investigated the complexity of the T‐cell receptor (TCR) repertoire in the prostate gland by examining the diversity of CD8+ TCR β chain variable region (Vβ) gene sequences in both the peripheral blood and prostates of cancer patients. Vβ repertoire analysis was performed by family‐specific Vβ spectratyping and flow cytometry, as well as direct sequence analysis (5′ RACE and cloning). Programmed cell death 1 (PD‐1 or PDCD1) expression on peripheral blood CD8+ T cells and CD8+ PIL was analyzed by flow cytometry.

RESULTS

CD8+ PIL isolated from cancer patients exhibited restricted TCR Vβ gene usage, and identical clones were identified in multiple sites within the prostate. Furthermore, CD8+ PIL express high levels of the inhibitory receptor PD‐1, a cell surface protein associated with an “exhausted” CD8+ T‐cell phenotype.

CONCLUSIONS

CD8+ PIL appear to have undergone clonal expansion in response to an as yet unidentified antigen; however, due to the high expression of PD‐1, these cells are likely incapable of mounting an effective immune response. The results provide an important basis for further efforts aimed at the identification of specific antigens involved in prostatic inflammation, and suggest that PD‐1 blockade may be useful in immunotherapy for PCa. Prostate 69: 1694–1703, 2009. © 2009 Wiley‐Liss, Inc.
Androgen receptor as a potential sign of prostate cancer metastasis
Authors: Lévesque Marie‐Hélène , El‐Alfy Mohamed , Cusan Leonello , Labrie Fernand
Published: August 2009

BACKGROUND

Androgen receptor (AR) expression and its modulation through the carcinogenesis process have been investigated in several studies with conflicting results.

MATERIALS AND METHODS

In situ hybridization and immunocytochemistry were used to examine AR expression in prostatic needle core biopsies of benign, high grade prostatic intraepithelial neoplasia (HGPIN) and prostatic adenocarcinoma.

RESULTS

A significant increase in AR mRNA levels was found in the cancerous prostatic cells when compared with the benign tissue biopsies. AR abundance in HGPIN was found to be almost half‐way between that observed in benign and in cancerous tissue. In the benign prostatic epithelium, the immunocytochemistry data show that AR is exclusively expressed in the nuclei of epithelial cells. However, in 72% of examined cancer biopsies, AR was expressed in both the cytoplasm and nuclei. After examination of medical records of 100 patients diagnosed with prostate cancer, it was found that the AR was expressed in both cellular compartments of cancer cells in 81% of cases when cancer was found to have metastasized outside the prostate. In contrast, when the cancer was organ‐confined, AR was localized in both the nuclei and cytoplasm in only 66% of cases. Moreover, when the AR was expressed in the cytoplasm of cancerous cells, consecutive serial sections immunostained with the mitochondrial marker suggest that AR is localized in the mitochondria.

CONCLUSIONS

AR mRNA expression is significantly higher in prostate cancer when compared to benign prostatic tissue. Prostate 69: 1704–1711, 2009. © 2009 Wiley‐Liss, Inc.
Inflammation in benign prostatic hyperplasia: A 282 patients' immunohistochemical analysis
Authors: Robert Grégoire , Descazeaud Aurélien , Nicolaïew Nathalie , Terry Stéphane , Sirab Nanor , Vacherot Francis , Maillé Pascale , Allory Yves , de la Taille Alexandre
Published: August 2009

INTRODUCTION AND OBJECTIVES

Prostatic inflammation could be a key component in prostate enlargement and benign prostatic hyperplasia (BPH) progression. Our aim was to characterize inflammatory cells infiltrate within BPH tissue and to correlate inflammation and clinical data.

MATERIAL AND METHODS

Inflammation was profiled on three clinical outcome tissue microarrays (TMAs), including 282 patients treated by surgery for a complicated and/or symptomatic BPH. Inflammation score was defined by combining six cytological parameters and five markers on immunohistochemistry (IHC). Cytological parameters were lymphocytes, macrophages, and polynuclears leukocytes infiltrates, and three glandular aspect modifications: glandular atrophy, glandular destruction, and tissue fibrosis. IHC markers were CD3, CD4, and CD8 decorating T‐lymphocytes, CD20 decorating B‐lymphocytes, and CD163 decorating macrophages.

RESULTS

The majority of patients had inflammatory cells infiltrating BPH tissues: 81% had T‐lymphocytes markers (CD3), 52% had B‐lymphocytes markers (CD20), and 82% had macrophages markers (CD163). IPSS score (21 vs. 12; P = 0.02) and prostate volume (77 cm3 vs. 62 cm3; P = 0.002) were significantly higher in patients with high‐grade prostatic inflammation.

CONCLUSION

We characterized inflammatory cells infiltrate in a large cohort of surgically treated BPH specimens. The role of inflammation in BPH development was highlighted by the strong correlation between histological inflammation, IPSS, and prostate volume. Prostate enlargement due to chronic inflammatory process may progressively conduce to BPH progression. Therefore, inflammation is a therapeutic target for BPH. Prostate 69: 1774–1780, 2009. © 2009 Wiley‐Liss, Inc.
Prostasin regulates iNOS and cyclin D1 expression by modulating protease‐activated receptor‐2 signaling in prostate epithelial cells
Authors: Chen Li‐Mei , Hatfield Meghan L. , Fu Ya‐Yuan , Chai Karl X.
Published: August 2009

BACKGROUND

Prostasin is down‐regulated during inflammation and in invasive cancers, and plays a role in regulation of inflammatory gene expression and invasion.

METHODS

We used the human benign prostatic hyperplasia cell line BPH‐1 to investigate gene expression changes associated with siRNA‐mediated loss of prostasin expression. Quantitative PCR and/or western blotting were used to evaluate the expression changes of iNOS, ICAM‐1, cyclin D1, IL‐6, and IL‐8. Gene expression changes were also evaluated in the presence of a PAR‐2 antagonist. The PC‐3 human prostate cancer cell line was used for evaluation of gene expression in response to prostasin re‐expression.

RESULTS

Prostasin silencing in BPH‐1 was associated with up‐regulation of iNOS, ICAM‐1, IL‐6, and IL‐8, and down‐regulation of cyclin D1; as well as reduced proliferation and invasion. The iNOS up‐regulation and cyclin D1 down‐regulation associated with prostasin silencing were inhibited by a PAR‐2 antagonist. Re‐expression of prostasin, a serine active‐site mutant, and a GPI‐anchor‐free mutant, in the PC‐3 cells resulted in PAR‐2 and cyclin D1 transcription up‐regulation. Transcription up‐regulation of IL‐6 and IL‐8 was associated with re‐expression of the serine active‐site mutant prostasin in the PC‐3 cells. Transcription up‐regulation of IL‐8, but to a lesser extent, was also observed in PC‐3 cells expressing the wild‐type prostasin. Expression of a serine protease active prostasin, GPI‐anchored or anchor‐free, prevented the IL‐6 induction in response to PAR‐2. The GPI‐anchor‐free prostasin also prevented the IL‐8 induction.

CONCLUSIONS

Prostasin plays a negative regulatory role on PAR‐2‐mediated signaling in prostate epithelial cells. Prostate 69: 1790–1801, 2009. © 2009 Wiley‐Liss, Inc.