bluuebunny/crossref_metadata_2025 huggingface dataset:
New silicone hydrogels based on interpenetrating polymer networks comprising polysiloxane and poly(vinyl alcohol) networks
Authors: Pavlyuchenko V. N. , Sorochinskaya O. V. , Ivanchev S. S. , Khaikin S. Ya. , Trounov V. A. , Lebedev V. T. , Sosnov E. A. , Gofman I. V.
Published: January 2009

A method for the synthesis of a new silicone hydrogel as a biphase material for soft contact lenses is considered. The method is based on the synthesis of sequential interpenetrating polymer networks (IPN) and includes the following stages: (1) cross‐linked silicone synthesis by the reaction of vinyl‐ and hydride‐containing oligosiloxanes; (2) silicone network saturation with vinyl acetate and cross‐linking monomer followed by UV‐initiated polymerization to form an IPN comprising the silicone and cross‐linked poly(vinyl acetate) (PVAc) network; (3) PVAc network alcoholysis with methanol to obtain silicone hydrogels comprising the silicone and cross‐linked poly(vinyl alcohol) (PVAl). A study of hydrophilic, optical, mechanical, and structural features of the silicone hydrogels showed that optical transparency is achieved for materials with the highest density of silicone network cross‐linking where the size of IPN structural units does not exceed 100 nm. The water content in hydrophilic networks of silicone hydrogel is found to be below the values typical of cross‐linked PVAl, leading to non‐additivity of IPN mechanical properties. Indeed, the elasticity moduli (E) of the hydrophilic and silicone networks are 0.4–0.7 and 0.7–1.8 MPa, respectively, whereas for some IPN this value reaches 3.0 MPa. The optimal parameters of synthesis providing the reduction of E to 0.8–1.6 MPa without deterioration of the required performance characteristics (optical transparency 90–92%, water content 20–39 wt%) are determined. Copyright © 2009 John Wiley & Sons, Ltd.
Thermosensitive polylactide‐glycolide delivery systems for treatment of narcotic addictions
Authors: Salehi R. , Nowruzi K. , Entezami A. A. , Asgharzadeh V. , Davaran S.
Published: January 2009

Environmental switches may be fabricated for the controlled release of pharmaceutical drug using a thermally responsive polymer with the intrinsic chemical and physical nature of stimuli‐sensitive smart materials. Particularly, much attention has been paid to the biomedical applications of poly(N‐isopropyl acrylamide) (PNIPAAm) because of its unique reversible transition at a specific lower critical solution temperature (LCST).Thermally sensitive block copolymers, poly(N‐isopropyl acrylamide‐b‐poly(L‐lactide‐co‐glycolide) (PNIPAAm‐b‐PLGA), and polyethylene glycol‐poly (lactide‐co‐glycolide) (PEG‐PLGA) triblock copolymers with different compositions and length of PLGA block were synthesized via ring‐opening polymerization of lactide and glycolide in the presence of OH‐terminated PNIPAAm or PEG. The composition and structure of the polymer were determined by NMR and FTIR. The effect of important factors, such as ionic strength, pH, and polymer concentration on the phase transition behavior of temperature‐sensitive polymers, were investigated by cloud point measurements. The resulting thermosensitive polymers were used for the entrapment of a narcotic antagonist drug, naltrexone, as the model drug. The loading efficiency and drug release behavior of naltrexone‐loaded hydrogels were investigated. The naltrexone loaded thermosensitive polymers were able to sustain the release of naltrexone for different periods of time, depending on the polymer composition, and concentration. In vitro release studies showed that these thermosensitive polymers are able to deliver naltrexone in biologically active forms at a controlled rate for 3–8 weeks. Copyright © 2009 John Wiley & Sons, Ltd.
Removal of lipopolysaccharide and reactive oxygen species using sialic acid immobilized polysulfone dialyzer
Authors: Chang Jung‐Jhih , Lin Po‐Ju , Yang Ming‐Chien , Chien Chiang‐Ting
Published: January 2009

Sialic acid (N‐acetylneuraminic acid, NANA) was covalently immobilized onto the surface of a polysulfone (PSF) hollow fiber membrane. Prior to the immobilization, the surface of PSF was treated with ozone, followed by grafting with acrylic acid, and then the esterification of NANA. The surface concentration of NANA was determined by 2‐thiobarbituric acid (TBA) test. Hemocompatibility, the capability of suppressing oxidative stress, and clearance of lipopolysaccharide (LPS) from the resulting hollow fiber membrane were evaluated. The results show that by immobilizing NANA onto PSF hollow fiber, the adhesion of platelet was reduced, while both APTT and PT were little affected. Furthermore, oxidative stress was suppressed by NANA‐immobilized PSF hollow fibers. The level of LPS was also greatly reduced. Copyright © 2009 John Wiley & Sons, Ltd.
Novel AB crosslinked polymer networks from telechelic 4‐vinylbenzyl carbamate terminated polyurethanes and different vinyl monomers
Authors: Venkataramani Sriram , Kannan Tharanikkarasu , Das Palash Jyoti , Radhakrishnan Ganga
Published: January 2009

Novel AB crosslinked polymer (ABCP) networks were synthesized from telechelic 4‐vinylbenzyl carbamate terminated polyurethanes and monomers such as styrene, 4‐vinylpyridine, methyl methacrylate and butyl acrylate. Telechelic 4‐vinylbenzyl carbamate terminated polyurethanes were synthesized from polypropylene glycol‐based NCO‐terminated polyurethane and vinylbenzyl alcohol. Effect of changing the molecular weight of polypropylene glycol on the static and dynamic mechanical properties of ABCP networks from polyurethane‐polymethyl methacrylate was studied in detail. Dynamic mechanical thermal analysis results show that polymethyl methacrylate and polystyrene‐based ABCPs have good damping over a broad temperature range. ABCP networks prepared from 4‐vinylbenzyl carbamate terminated polyurethane and different monomers such as methyl methacrylate, butyl acrylate and styrene exhibit single tan δmax value which implies excellent interlocking between the two polymers present in the ABCP networks. Static mechanical studies showed that methyl methacrylate and styrene‐based ABCP networks exhibit better tensile properties compared to other ABCP networks from butyl acrylate and 4‐vinyl pyridine monomers. Thermogravimetric analysis results revealed that the ABCP networks showed an improved thermal stability. Copyright © 2009 John Wiley & Sons, Ltd.
Mechanisms of lymphatic metastasis in human colorectal adenocarcinoma
Authors: Royston Daniel , Jackson David G.
Published: January 2009

The invasion of lymphatic vessels by colorectal cancer (CRC) and its subsequent spread to draining lymph nodes is a key determinant of prognosis in this common and frequently fatal malignancy. Although tumoural lymphangiogenesis is assumed to contribute to this process, review of the current literature fails to support any notion of a simple correlation between lymphatic vessel density and CRC metastasis. Furthermore, attempts to correlate the expression of various lymphangiogenic growth factors, most notably VEGF‐C and VEGF‐D, with the lymphatic metastasis of CRC have provided contradictory results. Recent evidence from animal and human models of tumour metastasis suggests that complex functional and biochemical interactions between the microvasculature of tumours and other cell types within the tumour microenvironment may play a pivotal role in the behaviour of commonly metastasizing tumours. Indeed, previous insights into tumoural blood vessels have provided candidate markers of tumoural angiogenesis that are currently the subject of intense investigation as future therapeutic targets. In this review article we survey the current evidence relating lymphangiogenesis and lymphangiogenic growth factor production to metastasis by CRC, and attempt to provide some insight into the apparent discrepancies within the literature. In particular, we also discuss some new and provocative insights into the properties of tumoural lymphatics suggesting that they have specific expression profiles distinct from those of normal lymphatic vessels and that appear to promote metastasis. These findings raise the exciting prospect of future biomarkers of lymphatic metastasis and identify potential targets for new generation anti‐tumour therapies. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Novel concepts in atherogenesis: angiogenesis and hypoxia in atherosclerosis
Authors: Sluimer Judith C , Daemen Mat J
Published: January 2009

The clinical complications of atherosclerosis are caused by thrombus formation, which in turn results from rupture of an unstable atherosclerotic plaque. The formation of microvessels (angiogenesis) in an atherosclerotic plaque contributes to the development of plaques, increasing the risk of rupture. Microvessel content increases with human plaque progression and is likely stimulated by plaque hypoxia, reactive oxygen species and hypoxia‐inducible factor (HIF) signalling. The presence of plaque hypoxia is primarily determined by plaque inflammation (increasing oxygen demand), while the contribution of plaque thickness (reducing oxygen supply) seems to be minor. Inflammation and hypoxia are almost interchangeable and both stimuli may initiate HIF‐driven angiogenesis in atherosclerosis. Despite the scarcity of microvessels in animal models, atherogenesis is not limited in these models. This suggests that abundant plaque angiogenesis is not a requirement for atherogenesis and may be a physiological response to the pathophysiological state of the arterial wall. However, the destruction of the integrity of microvessel endothelium likely leads to intraplaque haemorrhage and plaques at increased risk for rupture. Although a causal relation between the compromised microvessel structure and atherogenesis or between angiogenic stimuli and plaque angiogenesis remains tentative, both plaque angiogenesis and plaque hypoxia represent novel targets for non‐invasive imaging of plaques at risk for rupture, potentially permitting early diagnosis and/or risk prediction of patients with atherosclerosis in the near future. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Injury‐induced sequential transformation of notochordal nucleus pulposus to chondrogenic and fibrocartilaginous phenotype in the mouse
Authors: Yang Fan , Leung Victor YL , Luk Keith DK , Chan Danny , Cheung Kenneth MC
Published: January 2009

Intervertebral disc degeneration has been widely studied in different animal models. To test the hypothesis that needle puncture could induce progressive biochemical and molecular changes in murine discs, we established a mouse tail model to investigate the pathogenesis and molecular mechanism of puncture‐induced disc degeneration. Caudal discs in mouse tails were punctured using a 31G gauge needle at controlled depth under microscopic guidance. The progress of the disc degeneration was evaluated by radiographic analysis of disc height, histological grading and glycosaminoglycan (GAG) quantification pre‐operation and 1, 2, 6 and 12 weeks post‐puncture. Gene and protein expression of the extracellular matrix (ECM) was analysed by RT–PCR, in situ hybridization and immunohistochemistry. Histological study and disc height analysis revealed progressive degenerative changes in the punctured discs. Compared with the pre‐operation control group, total GAG content decreased 40% (p < 0.05) and aggrecan (Acan), decorin (Dcn) and versican (Vcan; Cspg2) expression was down‐regulated at 12 weeks post‐puncture. A transient increase of Col2a1‐expressing cells and elevation of collagen II protein in the nucleus pulposus (NP) was detected. Fibronectin (Fn1) expression was up‐regulated 50% and deposition of collagen I in NP was observed at 12 weeks post‐puncture. This study is the first to use an injury‐induced model to study disc degeneration in mouse. The disc degeneration involves a transient transformation of NP from notochordal to chondrogenic and eventually into fibrocartilaginous phenotype. The degenerative changes have some similarity to human disc degeneration, suggesting that this model may potentially be used in future to study the molecular mechanism and dissect the pathways of disc degeneration. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Current smoking‐specific gene expression signature in normal bronchial epithelium is enhanced in squamous cell lung cancer
Authors: Boelens Mirjam C , van den Berg Anke , Fehrmann Rudolf SN , Geerlings Marie , de Jong Wouter K , te Meerman Gerard J , Sietsma Hannie , Timens Wim , Postma Dirkje S , Groen Harry JM
Published: January 2009

Cigarette smoking is the main risk factor for the development of squamous cell lung carcinoma (SCC). However, the smoking‐related molecular changes in SCC have not been studied. Gene expression studies in both histologically normal bronchial epithelium and SCC epithelial samples identified genes differentially expressed between current and ex‐smokers. Subsequently, expression levels of the smoking‐related genes in normal bronchial epithelium were compared with those in SCC cells, since we hypothesized that the smoking‐induced changes would be also deregulated in SCC. Gene expression profiles were generated using Agilent whole human genome microarrays on laser‐microdissected normal bronchial epithelium and SCC samples. Expression levels of 246 genes, mainly related to oxidative stress response, were significantly different between normal bronchial epithelium of current and ex‐smokers. Such a differential gene expression profile did not exist in SCC cells of smokers and ex‐smokers. Interestingly, when comparing SCC and normal bronchial epithelium from ex‐smokers, the vast majority of these 246 genes were also deregulated in SCC. When comparing SCC with normal epithelium from smokers, 22% of the up‐regulated genes showed a similar high expression in SCC whereas 79% of the down‐regulated genes were even further reduced in SCC as compared to current smokers. The down‐regulated genes included several tumour suppressor genes, such as C9orf9, INHBB, LRIG1, SCGB3A1, SERPINI2, STEAP3 and ZMYND10. Thus, our study shows that the majority of genes up‐regulated in normal bronchial epithelium of current smokers show similar high expression levels in SCC, while down‐regulated genes are even further repressed in SCC. Our data indicate that smoking‐related changes in normal bronchial epithelial cells persist in malignant transformed squamous cells. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Human testicular (non)seminomatous germ cell tumours: the clinical implications of recent pathobiological insights
Authors: Looijenga Leendert HJ
Published: January 2009

Human germ cell tumours (GCTs) comprise several types of neoplasias with different pathogeneses and clinical behaviours. A classification into five subtypes has been proposed. Here, the so‐called type II testicular GCTs (TGCTs), ie the seminomas and non‐seminomas, will be reviewed with emphasis on pathogenesis and clinical implications. Various risk factors have been identified that define subpopulations of men who are amenable to early diagnosis. TGCTs are omnipotent, able to generate all differentiation lineages, both embryonic and extra‐embryonic, as well as the germ cell lineage itself. The precursor lesion, composed of primordial germ cells/gonocytes, is referred to as carcinoma in situ of the testis (CIS) and gonadoblastoma of the dysgenetic gonad. These pre‐malignant cells retain embryonic characteristics, which probably explains the unique responsiveness of the derived tumours to DNA‐damaging agents. Development of CIS and gonadoblastoma is crucially dependent on the micro‐environment created by Sertoli cells in the testis, and granulosa cells in the dysgenetic gonad. OCT3/4 has high sensitivity and specificity for CIS/gonadoblastoma, seminoma, and embryonal carcinoma, and is useful for the detection of CIS cells in semen, thus a promising tool for non‐invasive screening. Overdiagnosis of CIS due to germ cell maturation delay can be avoided using immunohistochemical detection of stem cell factor (SCF). Immunohistochemistry is helpful in making the distinction between seminoma and embryonal carcinoma, especially SOX17 and SOX2. The different non‐seminomatous histological elements can be recognized using various markers, such as AFP and hCG, while others need confirmation. The value of micro‐satellite instability as well as BRAF mutations in predicting treatment resistance needs validation in prospective trials. The availability of representative cell lines, both for seminoma and for embryonal carcinoma, allows mechanistic studies into the initiation and progression of this disease. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Latissimus dorsi muscle‐flap over Gore‐Tex patch for coverage of large thoracic defects in paediatric Ewing sarcoma
Authors: Gapany Christophe , Raffoul Wassim , Zambelli Pierre‐Yves , Joseph Jean‐Marc
Published: January 2009

Primary rib involvement accounts for 16% of paediatric Ewing sarcoma (ES). Neo‐adjuvant chemotherapy and surgical tumor resection may leave large thoracic wall defects requiring complex reconstruction in a growing individual. We report our experience in three children aged 3, 10, and 12 years, in whom single‐stage resection and reconstruction were performed using a Gore‐Tex Dualmesh patch, covered by a latissimus dorsi rotation flap harvested in continuity with the thoracolumbar fascia. The youngest patient also had a vertical expandable prosthetic titanium rib (VEPTR) anchored to help prevent subsequent scoliosis throughout growth. Pediatr Blood Cancer 2009;52:679–681. © 2009 Wiley‐Liss, Inc.
Simultaneous occurrence of MLL and RARA rearrangements in a pediatric acute lymphoblastic leukemia patient
Authors: Zámečníkova Adriana , Al Bahar Soad
Published: January 2009

We report a case of concurrent translocations of MLL gene, associated with a highly distinct leukemia subtype and RARA gene, which is pathogenomic in acute promyelocytic leukemia. Conventional cytogenetic revealed a novel complex rearrangement between chromosomes 5, 11, and 17 resulting in a three‐way chromosome translocation t(5;11;17)(q31;q23;q21). Fluorescence in situ hybridization analysis demonstrated that the 11q23 breakpoint involved the MLL, and the 17q21 breakpoint involved the RARA gene. Concurrent translocations of two specific oncogenes MLL and RARA with a new partner breakpoint on 5q31 have not been previously described. Pediatr Blood Cancer 2009;52:671–674. © 2009 Wiley‐Liss, Inc.
Response to high‐dose ifosfamide in patients with advanced/recurrent Ewing sarcoma
Authors: Ferrari S. , del Prever A. Brach , Palmerini E. , Staals E. , Berta M. , Balladelli A. , Picci P. , Fagioli F. , Bacci G. , Vanel D.
Published: January 2009

Aim

To determine activity and toxicity of high‐dose ifosfamide (HDIFO) in recurrent or advanced Ewing sarcoma family tumors (EFT).

Methods

Thirty‐seven EFT patients [median age 17 years (6–45 years)] previously treated with chemotherapy regimens including standard dose ifosfamide were enrolled. HDIFO was administered for metastatic recurrent disease in 33 patients and for progression during neoadjuvant chemotherapy in 4 patients. All patients who received two courses of 15 g/m2 ifosfamide were evaluable for radiographic response assessed according to RECIST criteria.

Results

Transient Grade 4 neutropenia and thrombocytopenia in 97% and 54% HDIFO courses respectively and severe CNS toxicity in one patient were observed. Thirty‐five patients were evaluable: 12 (34%) had complete (2) or partial (10) response, 11 (32%) had stable disease, and 12 (34%) had progression.

Conclusions

In patients with relapsed or advanced EFT previously treated with standard dose ifosfamide HDIFO is active and it should be considered a treatment option. Pediatr Blood Cancer 2009;52:581–584. © 2009 Wiley‐Liss, Inc.
Development of eastern blotting technique for sennoside A and sennoside B using anti‐sennoside A and anti‐sennoside B monoclonal antibodies
Authors: Morinaga Osamu , Uto Takuhiro , Sakamoto Seiichi , Putalun Waraporn , Lhieochaiphant Sorasak , Tanaka Hiroyuki , Shoyama Yukihiro
Published: January 2009

Introduction

Rhubarb, senna and sennoside‐containing preparations are currently widely employed as purgatives. The major active components of these medications are sennoside A (SA) and sennoside B (SB).

Objective

To develop an eastern blotting technique for the specific visualisation and easy determination of SA and SB in plant extracts for application in the standardisation and authentication of rhubarb and senna.

Methodology

SA and SB were separated by TLC, transferred to a PVDF membrane, treated with 1‐ethyl‐3‐(3′‐dimethylaminopropyl)‐carbodiimide hydrochloride solution and finally treated with bovine serum albumin (BSA). The resulting membrane‐bound SA–BSA and SB–BSA conjugates were linked to anti‐SA and anti‐SB monoclonal antibodies (MAbs) and then to secondary antibodies labelled with peroxidase. SA and SB were detected by visualisation of the peroxidase reaction products.

Results

The limit of detection of the eastern blotting was 62.5 ng for both sennosides. The method was applied to the immunohistochemical localisation of SA in fresh rhubarb root. Phloem and radiate wood were found to contain higher concentrations of SA compared with other tissues (pith and bud) in agreement with results obtained by ELISA. The concentrations of SA in the phloem, radiate wood, pith and bud were 64.4, 48.1, 15.0 and 1.8 ng/mg fresh weight, respectively.

Conclusion

The technique described permitted the visualisation of small molecular weight compounds that had been bound to a membrane, using immunostaining. Owing to the specificity of the MAbs, the eastern blotting may prove to be a useful method for the identification of SA and SB in a background containing large amount of impurities. Copyright © 2009 John Wiley & Sons, Ltd.
Comparative proteomic analysis of human mesenchymal and embryonic stem cells: Towards the definition of a mesenchymal stem cell proteomic signature
Authors: Roche Stephane , Delorme Bruno , Oostendorp Robert A. J. , Barbet Romain , Caton David , Noel Daniele , Boumediene Karim , Papadaki Helen A. , Cousin Beatrice , Crozet Carole , Milhavet Ollivier , Casteilla Louis , Hatzfeld Jacques , Jorgensen Christian , Charbord Pierre , Lehmann Sylvain
Published: January 2009

Mesenchymal stem cells (MSC) are adult multipotential progenitors which have a high potential in regenerative medicine. They can be isolated from different tissues throughout the body and their homogeneity in terms of phenotype and differentiation capacities is a real concern. To address this issue, we conducted a 2‐DE gel analysis of mesenchymal stem cells isolated from bone marrow (BM), adipose tissue, synovial membrane and umbilical vein wall. We confirmed that BM and adipose tissue derived cells were very similar, which argue for their interchangeable use for cell therapy. We also compared human mesenchymal to embryonic stem cells and showed that umbilical vein wall stem cells, a neo‐natal cell type, were closer to BM cells than to embryonic stem cells. Based on these proteomic data, we could propose a panel of proteins which were the basis for the definition of a mesenchymal stem cell proteomic signature.
Form follows function: Shape analysis of protein cavities for receptor‐based drug design
Authors: Weisel Martin , Proschak Ewgenij , Kriegl Jan M. , Schneider Gisbert
Published: January 2009

Identification of potential ligand‐binding pockets is an initial step in receptor‐based drug design. While many geometric or energy‐based binding‐site prediction methods characterize the size and shape of protein cavities, few of them offer an estimate of the pocket's ability to bind small drug‐like molecules. Here, we present a shape‐based technique to examine binding‐site druggability from the crystal structure of a given protein target. The method includes the PocketPicker algorithm to determine putative binding‐site volumes for ligand‐interaction. Pocket shape descriptors were calculated for both known ligand binding sites and empty pockets and were then subjected to self‐organizing map clustering. Descriptors were calculated for structures derived from a database of representative drug‐protein complexes with experimentally determined binding affinities to characterize the “druggable pocketome”. The new method provides a means for selecting drug targets and potential ligand‐binding pockets based on structural considerations and addresses orphan binding sites.
New iodo‐acetamido cyanines for labeling cysteine thiol residues. A strategy for evaluating plasma proteins and their oxido‐redox status
Authors: Bruschi Maurizio , Grilli Stefano , Candiano Giovanni , Fabbroni Serena , Della Ciana Leopoldo , Petretto Andrea , Santucci Laura , Urbani Andrea , Gusmano Rosanna , Scolari Francesco , Ghiggeri Gian Marco
Published: January 2009

Two new iodoacetamide‐substituted cyanines, C3NIASO3 and C5NIASO3, were synthesized starting from hemicyanine and were utilized for labeling plasma proteins. Specificity, sensitivity and feasibility for SH residues was tested utilizing an equimolar mixture of standard proteins and with normal plasma. Oxidized plasma proteins following H2O2 exposure and plasma from patients with focal glomerulosclerosis were analyzed as models of altered protein oxido‐redox status. Following optimization of the assay (dye/protein ratio, pH), C3NIASO3 and C5NIASO3 gave a sensitivity slightly better than N‐hydroxysuccinimidyl dyes for plasma proteins and were successfully employed for differential display electrophoresis (DIGE). Twenty‐nine proteins were detected in normal plasma after 2‐DE while less proteins were detected in plasma of patients with glomerulosclerosis. Following massive ‘in vitro' oxidation with H2O2, C3NIASO3 and C5NIASO3 failed to detect any residual SH, implicating massive oxidation. In conclusion, this study describes the synthesis of two new iodoacetamide cyanines that can be utilized for the analysis of plasma proteins with 2‐DE and DIGE. They are also indicated for the definition of the oxido‐redox status of proteins and were successfully utilized to extend the analysis of oxidation damage in patients with glomerulosclerosis.
In vivo phosphorylation sites of barley tonoplast proteins identified by a phosphoproteomic approach
Authors: Endler Anne , Reiland Sonja , Gerrits Bertran , Schmidt Ulrike G. , Baginsky Sacha , Martinoia Enrico
Published: January 2009

In plants the vacuolar functions are the cellular storage of soluble carbohydrates, organic acids, inorganic ions and toxic compounds. Transporters and channels located in the vacuolar membrane, the tonoplast, are modulated by PTMs to facilitate the optimal functioning of a large number of metabolic pathways. Here we present a phosphoproteomic approach for the identification of in vivo phosphorylation sites of tonoplast (vacuolar membrane) proteins. Highly purified tonoplast and tonoplast‐enriched microsomes were isolated from photosynthetically induced barley (Hordeum vulgare) mesophyll protoplasts. Phosphopeptides were enriched by strong cation exchange (SCX) chromatography followed either by IMAC or titanium dioxide (TiO2) affinity chromatography and were subsequently analysed using LC‐ESI‐MS/MS. In total, 65 phosphopeptides of 27 known vacuolar membrane proteins were identified, including the two vacuolar proton pumps, aquaporins, CAX transporters, Na+/H+ antiporters as well as other known vacuolar transporters mediating the transfer of potassium, sugars, sulphate and malate. The present study provides a novel source to further analyse the regulation of tonoplast proteins by protein phosphorylations, especially as most of the identified phosphorylation sites are highly conserved between Hordeum vulgare (Hv) and Arabidopsis thaliana.
Profiling of T‐cell receptor signaling complex assembly in human CD4 T‐lymphocytes using RP protein arrays
Authors: Brembilla Nicolò Costantino , Cohen‐Salmon Isabelle , Weber Johann , Rüegg Curzio , Quadroni Manfredo , Harshman Keith , Doucey Marie‐Agnès
Published: January 2009

The RP protein (RPP) array approach immobilizes minute amounts of cell lysates or tissue protein extracts as distinct microspots on NC‐coated slide. Subsequent detection with specific antibodies allows multiplexed quantification of proteins and their modifications at a scale that is beyond what traditional techniques can achieve. Cellular functions are the result of the coordinated action of signaling proteins assembled in macromolecular complexes. These signaling complexes are highly dynamic structures that change their composition with time and space to adapt to cell environment. Their comprehensive analysis requires until now relatively large amounts of cells (>5×107) due to their low abundance and breakdown during isolation procedure. In this study, we combined small scale affinity capture of the T‐cell receptor (TCR) and RPP arrays to follow TCR signaling complex assembly in human ex vivo isolated CD4 T‐cells. Using this strategy, we report specific recruitment of signaling components to the TCR complex upon T‐cell activation in as few as 0.5 million of cells. Second‐ to fourth‐order TCR interacting proteins were accurately quantified, making this strategy specially well‐suited to the analysis of membrane‐associated signaling complexes in limited amounts of cells or tissues, e.g., ex vivo isolated cells or clinical specimens.
Proteomic analysis of hepatic ischemia/reperfusion injury and ischemic preconditioning in mice revealed the protective role of ATP5β
Authors: Xu Chengfu , Zhang Xuequn , Yu Chaohui , Lu Guohua , Chen Shaohua , Xu Liming , Ding Wei , Shi Qiaojuan , Li Youming
Published: January 2009

Hepatic ischemia/reperfusion (I/R) injury is an inevitable consequence during liver surgery. Ischemic preconditioning (IPC) has been shown to protect the livers from I/R injury, partially mediated by preservation of hepatic ATP contents. However, the precise molecular mechanisms of these events remain poorly elucidated. In this study, liver proteomes of the mice subjected to I/R injury pretreated with or without IPC were analyzed using 2‐DE combined with MALDI‐TOF/TOF mass analysis. Twenty proteins showing more than 1.5‐fold difference were identified in the livers upon I/R injury. Among these proteins, four proteins were further regulated by IPC when compared with nonpretreated controls. One of these proteins, ATP synthase β subunit (ATP5β) catalyzes the rate‐limiting step of ATP formation. The expression level of ATP5β, which was further validated by Western blot analysis, was significantly decreased upon I/R injury while turned over by IPC pretreatment. Change pattern of hepatic ATP corresponded with that of ATP5β expression, indicating that increasing hepatic ATP5β expression might be a reason for ATP‐preserving effect of IPC. In summary, this study provided new clues for understanding the mechanisms of IPC against I/R injury. The protective role of ATP5β might give evidences for developing new therapeutic approaches against hepatic I/R injury.
Development of a screening tool for the identification of psychooncological treatment need in breast cancer patients
Authors: Meraner Verena , Giesinger Johannes , Kemmler Georg , Taucher Susanne , Hubalek Michael , Weber Barbara , Rumpold Gerhard , Sperner‐Unterweger Barbara , Holzner Bernhard
Published: January 2009

Objective: In breast cancer patients the diagnosis, treatment and aftercare of the physical disease cause a large amount of psychosocial distress, which can have a variety of negative consequences on patients' physical and mental well‐being. Although about one‐third of the patients show heightened psychosocial distress that may require psychooncological interventions its detection in daily clinical routine is poor and referral to mental health professionals is insufficient. The aim of the study was to develop a short screening tool for the detection of need for psychooncological treatment (POT) in breast cancer patients.

Methods: Over a period of six months, 115 breast cancer outpatients attending the Department of Gynaecology at Innsbruck Medical University were consecutively included in the study. Logistic regression analysis and ROC analyses were used to identify the most predictive item set from a set of questionnaires (EORTC‐QLQ‐C30, HADS and Hornheide Screening Instrument) and other additional questions.

Results: Data from 105 breast cancer patients (mean age 58.8, SD 12.3) were available for analysis. A logistic regression equation containing the EORTC‐QLQ‐C30 scales Emotional Functioning and Role Functioning as well as the yes–no question after psychiatric/psychological/psychotherapeutic treatment at any point in lifetime showed highest predictive power with regard to need of POT (AUC=0.88; CI 95% 0.82–0.95). A pilot study (n=20) investigating applicability of a computerized version of this screening tool in oncological routine showed high acceptance and feasibility.

Conclusion: The developed PO‐screening tool showed high diagnostic accuracy regarding POT needs. The short assessment time and good usability of its computerized version allowed easy implementation in daily oncological routine. Copyright © 2009 John Wiley & Sons, Ltd.
A cost effectiveness analysis of omitting radiography in diagnosis of acute bronchiolitis
Authors: Yong Jean Hai Ein , Schuh Suzanne , Rashidi Rasha , Vanderby Sonia , Lau Rodney , Laporte Audrey , Nauenberg Eric , Ungar Wendy J.
Published: January 2009

Objective

To carry out a cost‐effectiveness analysis of omitting chest radiography in the diagnosis of infant bronchiolitis.

Hypothesis

Omitting chest radiographs in the diagnosis of typical bronchiolitis was expected to reduce costs without adversely affecting the detection rate of alternate diseases.

Study Design

An economic evaluation was conducted using clinical and health resources. Emergency department (ED) physicians provided diagnoses pre‐ and post‐radiography as well as a management plan. The primary outcome was the diagnostic accuracy (false‐negative rate) of alternate diagnoses with and without X‐ray. The incremental costs of omitting radiography in comparison to routine radiography per patient were assessed from a health system perspective.

Patient Selection

We studied 265 infants, 2–23 months old, presenting at the ED with typical bronchiolitis. Patients with pre‐existing conditions or radiographs were omitted from the study.

Methodology

Expected costs to the health care system of including and excluding chest radiographs were compared, including costs associated with misdiagnosis.

Results

All alternate diagnoses (two cases) were missed by ED physicians pre‐ and post‐radiography, resulting in a 100% false negative rate. The specificity in detecting alternate diseases was 96.6% pre‐radiography and 88.6% post‐radiography. Of the 17 cases of coexistent pneumonia, 88% were missed pre‐radiography and 59% post‐radiography, with respective false positive rates of 10.5% and 16.1%. Omission of routine chest radiograph saved CDN $59 per patient, primarily due to savings in radiography and hospitalization costs. The economic benefit persisted after the inpatient length of stay, ED overhead and radiograph costs were varied.

Conclusion

For infants with typical bronchiolitis, omitting radiography is cost saving without compromising diagnostic accuracy of alternate diagnoses and of associated pneumonia. Pediatr Pulmonol. 2009; 44:122–127. © 2009 Wiley‐Liss, Inc.
Epidemiology and impacts of children hospitalized with pneumonia from 1997 to 2004 in Taiwan
Authors: Wu Ping‐Sheng , Chang I‐Shou , Tsai Fang‐Yu , Hsieh Yu‐Chia , Shao Pei‐Lan , Chang Luan‐Yin , Huang Li‐Min
Published: January 2009

Background

Few studies have investigated the epidemiology and disease burden of childhood pneumonia in Taiwan, a middle‐income country.

Methods

Using Taiwan's National Health Insurance database, we analyzed the annual population‐based incidence and mortality rate of hospitalizations due to pneumonia in children under 18 years old from 1997 to 2004 in Taiwan.

Results

The average annual incidence of hospitalized pneumonia was 1,240 episodes per 100,000 children in those below 18 years old. For children under 5 years old and for infants, the incidence was 3,965 and 4,984 out of 100,000, respectively. Boys were more likely to be affected than girls (male to female risk ratio 1.27, P < 0.001). The disease occurred most frequently in the spring and least frequently in the autumn (P < 0.001). In children below the age of 5 years old, mortality due to pneumonia was 6.7 per 100,000 per year for children and accounted for 4.2% of the total deaths in this population.

Conclusions

In Taiwan, pneumonia occurs most frequently in the spring, has an annual incidence of hospitalizations of 3,965 episodes per 100,000 children below the age of 5 years old. It accounts for a mortality rate of 6.7 per 100,000 children in that population, four times that of developed countries (1.6–1.8 episodes per 100,000 children‐year). Pediatr Pulmonol. 2009; 44:162–166. © 2009 Wiley‐Liss, Inc.
Barriers to asthma self‐management in adolescents: Relationships to psychosocial factors
Authors: Rhee Hyekyun , Belyea Michael J. , Ciurzynski Susan , Brasch Judith
Published: January 2009

Asthma morbidity in adolescents often results from inadequate asthma self‐management. This study was to explore barriers to self‐management perceived by adolescents and to examine the associations between barriers and psychosocial factors including knowledge, attitude and self‐efficacy. This cross‐sectional study included a total of 126 adolescents with asthma (13–20 years) representing diverse race/ethnicity groups with a wide range of socioeconomic status. Self‐reported data were analyzed using descriptive statistics, factor analysis and hierarchical regression. The most frequently endorsed barrier (63%) was adolescents' unwillingness to give up “the things the doctors say I have to give up,” followed by difficulty in remembering to take care of their asthma (53%), and then “trying to forget” that they have asthma (50%). Psychosocial factors accounted for 32% of the variance in total barrier perceptions. Factor analysis revealed barriers in four domains including negativity toward providers and the medication regimen, cognitive difficulty, peer/family influence and denial. Self‐efficacy was found to be the most influential factor that showed strong negative associations with all four barrier subscales independent of the levels of asthma control and sociodemographic characteristics. Poor attitudes toward asthma were also associated with barriers of cognitive difficulty and social influence after adjusting for other factors. Males consistently reported higher total barriers and barriers of negativity, social influence and denial. The gender differences were not explained by psychosocial and sociodemographic factors. This study suggests that psychosocial factors are strong predictors of barriers to self‐management in adolescents. Particularly, promoting self‐efficacy may be beneficial in addressing the barriers. Special attention is needed to address the higher propensity for barriers in males. Pediatr Pulmonol. 2009; 44:183–191. © 2009 Wiley‐Liss, Inc.
Quantification of phase I/II metabolizing enzyme gene expression and polycyclic aromatic hydrocarbon–DNA adduct levels in human prostate
Authors: John Kaarthik , Ragavan Narasimhan , Pratt M. Margaret , Singh Paras B. , Al‐Buheissi Salah , Matanhelia Shyam S. , Phillips David H. , Poirier Miriam C. , Martin Francis L.
Published: January 2009

BACKGROUND

Studies of migrant populations suggest that dietary and/or environmental factors play a crucial role in the etiology of prostatic adenocarcinoma (CaP). The human prostate consists of the peripheral zone (PZ), transition zone (TZ), and central zone (CZ); CaP occurs most often in the PZ.

METHODS

To investigate the notion that an underlying differential expression of phase I/II genes, and/or the presence of polycyclic aromatic hydrocarbon (PAH)–DNA adducts might explain the elevated PZ susceptibility, we examined prostate tissues (matched tissue sets consisting of PZ and TZ) from men undergoing radical retropubic prostatectomy for CaP (n = 26) or cystoprostatectomy (n = 1). Quantitative gene expression analysis was employed for cytochrome P450 (CYP) isoformsCYP1A1,CYP1B1, andCYP1A2, as well asN‐acetyltransferase 1 and 2 (NAT1andNAT2) and catechol‐O‐methyl transferase (COMT).

RESULTS

CYP1B1,NAT1, andCOMTwere expressed in all tissue sets; levels ofCYP1B1andNAT1were consistently higher in the PZ compared to TZ. Immunohistochemistry confirmed the presence of CYP1B1 (nuclear‐associated and primarily in basal epithelial cells) and NAT1. Normal tissue from 23 of these aforementioned 27 matched tissue sets was analyzed for PAH–DNA adduct levels using antiserum elicited against DNA modified withr,7t8‐dihydroxy‐t‐9,10‐oxy‐7,8,9,10‐tetrahydro‐benzo[a]pyrene (BPDE). PAH–DNA adduct levels were highest in glandular epithelial cells, but a comparison of PZ and TZ showed no significant differences.

CONCLUSION

Although expression of activating and/or detoxifying enzymes may be higher in the PZ, PAH–DNA adduct levels appear to be similar in both zones. Therefore, factors other than PAH–DNA adducts may be responsible for promotion of tumor formation in the human prostate. Prostate 69:505–519, 2009. © 2009 Wiley‐Liss, Inc.
Dietary fatty acid quality affects AR and PPARγ levels and prostate growth
Authors: Escobar Esdras L.O. , Gomes‐Marcondes Maria Cristina C. , Carvalho Hernandes F.
Published: January 2009

BACKGROUND

Fatty acids are among the most important nutritional factors associated with the ethiopathogenesis of prostate cancer, therefore the main objective of this work was to evaluate the effect of quality of fatty acid on the rat ventral prostate growth, tissue organization, and expression of androgen receptor (AR) and peroxisome proliferation activator receptor γ (PPARγ).

METHODS

Wistar rats were distributed into five groups, which were fed isocaloric normolipidic diets containing soybean oil (7% Control), linseed (7% or 3.5% linseed plus 3.5% soybean oil) and rendered pork fat (7% or 3.5% lard plus 3.5% soybean oil) for 10 weeks after weaning.

RESULTS

At the end of treatment, the experiments demonstrated that lard and linseed oil caused opposite effects on prostatic growth. While the lard promoted an increase in prostatic weight associated to epithelial hyperplasia (confirmed by stereology); the linseed resulted in a significantly lighter organ. Immunohistochemistry and Western blotting demonstrated increased expression of AR and PPARγ in groups fed with lard diet, while linseed oil promoted a decrease.

CONCLUSIONS

Prostatic growth is influenced by dietary fatty acids with concurrent variation in the expression of AR and PPARγ. PPARγ might represent the link between diet and prostate growth and AR expression and function. Since the levels of testosterone were altered it is also possible that prostatic changes are secondary to systemic effects of the diet. Prostate 69:548–558, 2009. © 2009 Wiley‐Liss, Inc.
Androgen ablation augments prostate cancer vaccine immunogenicity only when applied after immunization
Authors: Koh Yi T. , Gray Andrew , Higgins Sean A. , Hubby Bolyn , Kast W. Martin
Published: January 2009

BACKGROUND

Androgen ablation (AA) causes apoptosis of normal and neoplastic prostate cells. It is a standard treatment for advanced prostate cancer. Androgen ablation‐mediated immunological effects include bone marrow hyperplasia, thymic regeneration, T and B cell lymphopoeisis and restoration of age‐related peripheral T cell dysfunction. Androgens also regulate the transcription of several cytokines. Dendritic cells (DC) are the most potent antigen presenting cells that can activate antigen‐specific naïve T cells. Despite myriad clinical trials involving DC‐based prostate cancer immunotherapies, the effects of AA on DC function remain largely uncharacterized. Therefore, we investigated the effects of AA on DC and whether it could improve the efficacy of prostate cancer immunotherapy.

METHODS

Cytokine expression changes due to AA were quantified by multiplex ELISA. Flow cytometry was used to assess AA‐mediated effects on DC maturation and expression of costimulatory markers. Mixed leukocyte reactions and cell‐mediated lysis assays elucidated the role of androgens in DC function. The effect of AA on the efficacy of vaccination against a prostate tumor‐associated antigen was tested using Elispot assays.

RESULTS

Androgen ablation increased dendritic cell maturation and costimulatory marker expression, but had no effect on DC costimulatory function. However, DC isolated from castrated mice increased the expression of key cytokines by antigen‐experienced T cells while decreasing their expression in naïve cells. Finally, androgen ablation improved immune responses to vaccination only when applied after immunization.

CONCLUSION

Androgen ablation causes differential effects of DC on primary and secondary T cell responses, thus augmenting vaccine immunogenicity only when applied after immunization. Prostate 69:571–584, 2009. © 2009 Wiley‐Liss, Inc.
An updating meta‐analysis of the GSTM1, GSTT1, and GSTP1 polymorphisms and prostate cancer: A HuGE Review
Authors: Mo Zengnan , Gao Yong , Cao Yunfei , Gao Feng , Jian Lijuan
Published: January 2009

It has been postulated that individuals with GSTM1, GSTT1 deficiency and, GSTP1 (105Ile/Val transition) have increased susceptibility to carcinogens and are more likely to develop prostate cancer. In recent years, GST status has been extensively studied as a prostate cancer risk factor; however, the results are inconsistent. To re‐examine this controversy, we have undertaken an updating meta‐analysis of 29 studies with GSTM1 genotyping (4,564 prostate cancer cases and 5,464 controls), 22 studies with GSTT1 genotyping (3,837 cases and 4,552 controls), and 24 studies with GSTP1 genotyping (5,301 cases and 5,621 controls). The random effects odds ratio was 1.33 [95% confidence interval (95% CI): 1.15, 1.55; I2 = 68.9%, P for heterogeneity = 0.00] for the GSTM1 null versus present genotype and 1.05 (95% CI: 0.86, 1.27; I2 = 68.2%, P for heterogeneity = 0.00) for the GSTT1 null versus present genotype, and 1.06 (95% CI: 0.91, 1.24; I2 = 71.5%, P for heterogeneity = 0.00) for the GSTP1‐Val versus GSTP1‐Ile allele. For GSTM1 polymorphism, similar results reached in Caucasians and Asians, with exception for Africans. No association between GSTT1 or GSTP1 polymorphisms and prostate cancer risk was detected in different racial. In conclusion, the major finding of our study suggested that GSTM1 polymorphism conferred an increasing risk of prostate cancer on a wide population basis, however, no relationship was found between GSTT1 and GSTP1 status and the risk of prostate cancer. Prostate 69:662–688, 2009. © 2009 Wiley‐Liss, Inc.