bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of bisphosphonates for the treatment of osteonecrosis as a complication of therapy for childhood acute lymphoblastic leukaemia (ALL)
Authors: Kotecha Rishi S. , Powers Neil , Lee Senq‐J. , Murray Kevin J. , Carter Tina , Cole Catherine
Published: February 2010

Background

Osteonecrosis is a well‐recognised complication of current childhood acute lymphoblastic leukaemia (ALL) therapy. There are few studies on the medical management of osteonecrosis in this setting. We studied the therapeutic and radiological effects of oral and intravenous bisphosphonate use compared with standard care as treatment for osteonecrosis in this population.

Method

Patients who developed osteonecrosis as a complication of ALL therapy between 1994 and 2007 were treated at a single paediatric institution. Of 17 patients, 9 were commenced on bisphosphonates and 8 treated conservatively. Both groups were observed with time. Pain, analgesic requirement and musculoskeletal function were assessed monthly. Affected joints were radiologically imaged at set intervals. Each scan was graded using an ellipsoid method to give the total volume of osteonecrosis, by blinded radiologic examination.

Results

Three of six patients treated with oral alendronate showed clinical improvement. The three patients that had no improvement were subsequently treated with intravenous pamidronate. All six patients treated with intravenous pamidronate showed clinical improvement. Seven of eight conservatively treated patients deteriorated clinically. All patients demonstrated reduction in the radiological burden of osteonecrosis with time. There was no difference in the rate of reduction between conservative and bisphosphonate arms.

Conclusion

Bisphosphonate use, in particular pamidronate, improved pain scores, analgesic requirement and musculoskeletal function in patients with osteonecrosis occurring as a complication of childhood ALL therapy. Objective radiologic benefit of bisphosphonate treatment could not be demonstrated. Risks, benefits and long‐term outcome of bisphosphonate use in this population should be addressed in a larger prospective, randomised trial. Pediatr Blood Cancer 2010;54:934–940 © 2010 Wiley‐Liss, Inc.
Undifferentiated sarcoma of the thyroid in a child
Authors: Postovsky Sergey , Vlodavsky Eugene , Kuten Abraham , Shendler Yaakov , Doweck Ilana , Arush Myriam Weyl Ben
Published: February 2010

The most common malignant tumor of the thyroid is papillary carcinoma. Sarcoma of the thyroid is encountered very rarely; its therapy is complex and poses significant problems due to the problematic location of the tumor. A 14‐year‐old female was diagnosed with undifferentiated sarcoma of the thyroid and received combined therapy comprising surgery, chemo‐ and radiotherapy without significant side effects. This case underlines the fact that undifferentiated thyroid sarcoma may be a diagnostic possibility in children with malignant masses of the neck and may be successfully treated with modern therapeutic strategies. Pediatr Blood Cancer 2010;54:1038–1040 © 2010 Wiley‐Liss, Inc.
Quality of life in young patients after bone tumor surgery around the knee joint and comparison with healthy controls
Authors: Bekkering W. Peter , Vliet Vlieland Theodora P.M. , Koopman Hendrik M. , Schaap Gerard R. , Schreuder H.W. Bart , Beishuizen Auke , Tissing Wim J.E. , Hoogerbrugge Peter M. , Anninga Jacob K. , Taminiau Antonie H.M.
Published: February 2010

Background

This study aimed to compare the health related quality of life (HRQoL) of children and adolescents after malignant bone tumor surgery of the leg with healthy controls.

Procedure

Patients between 8 and 25 years old were cross‐sectional recruited. Patients under 16 years of age received the TNO (Netherlands Organization for Applied Scientific Research) and AZL (Leiden University Medical Center) Children's Quality of Life Questionnaire (TACQOL), patients aged 16 years and older received the TNO‐AZL Questionnaire for Adult's Quality of Life (TAAQOL) and the Short Form‐36 (SF‐36). Three age‐ and sex‐matched normative random samples, drawn from large, nationwide studies, were used for the comparison with healthy controls. Patients were interviewed regarding their most important problems related to the disease and its treatment.

Results

Eighty‐one patients with a mean age of 16.9 years (SD 4.2) were included (41 female). Limb sparing surgery was executed in 38 patients, ablative surgery in 43 patients. In comparison with healthy controls, patients had significantly poorer HRQoL within the domains autonomy and motor function of the TACQOL, gross motor function, cognitive functioning, daily functioning and sexuality of the TAAQOL, and physical functioning, role physical, general health, and the physical and mental component summary scales of the SF‐36. Patients reported limitations in physical activities, participation in sports, and cosmetic aspects as the most detrimental consequences of their disease and its treatment.

Conclusion

In children and adolescents who underwent surgery for a malignant tumor of the leg physical, functioning was significantly impaired as compared to healthy controls. Pediatr Blood Cancer 2010;54:738–745. © 2010 Wiley‐Liss, Inc.
Direct and rapid determination of ultratrace heavy metals in solid plant materials by ET‐AAS ultrasonic‐assisted slurry sampling
Authors: Sánchez‐Moreno Raúl A. , Gismera M. Jesús , Sevilla M. Teresa , Procopio Jesús R.
Published: February 2010

Introduction – Plants can be used as bioindicators in the study of contamination processes by heavy metals. Most of the analytical methodologies used for determination of metals in plants are based on atomic techniques with previous wet digestion of the solid samples. Methodologies that allow direct metal measurements in solid samples are very attractive alternatives.

Objective – To develop a new procedure for direct analysis of copper, nickel, cadmium and lead at very low concentration levels in leaves based on electrothermal atomic absorption spectroscopy (ET‐AAS) with introduction of samples as a slurry.

Methodology – In order to obtain accurate and precise results even at very low concentrations, the different parameters that influence the sample slurry preparation such as acid percentage, presence of stabilising agents and ultrasonic probe operation were studied. Instrumental parameters such as chemical modifier and temperature and times for drying, pyrolysis and atomisation steps that influence ET‐AAS measurement were optimised.

Results – Optimal slurry conditions for copper and nickel determination were 0.5% Tween 85 with 5% nitric acid. For lead and cadmium analysis the best results were obtained in 5% nitric acid without stabilising agents. The achieved detection limits were 0.023 mg/kg for copper, 0.018 mg/kg for nickel, 0.0002 mg/kg for cadmium and 0.009 mg/kg for lead. For validation purposes, the method was applied to metal analysis in a pine needles reference material.

Conclusion – According to our knowledge, the detection limits obtained are the best reported in the literature. The methodology was successfully used in metal determinations in actual leaf samples. Copyright © 2010 John Wiley & Sons, Ltd.
Fabrication of fluorinated polymer nanosheets using the Langmuir–Blodgett technique: characterization of their surface properties and applications
Authors: Aminuzzaman Mohammod , Mitsuishi Masaya , Miyashita Tokuji
Published: February 2010

This review describes the bottom‐up design of ultra‐thin fluorinated polymer films. Fluorinated polymer assemblies offer unique and application‐oriented properties strongly correlated with surface chemistry. We focus on our recent topic of research, fluorinated polymer nanosheets, which are ultra‐thin polymer films fabricated from amphiphilic fluorinated polyalkylacrylamides using the elegant Langmuir–Blodgett technique. These fluorinated polymer nanosheets have excellent hydrophobicity, extremely low surface energy and a low friction coefficient. Therefore, fluorinated polymer nanosheets are attractive nanomaterials for surface modification in terms of tailoring of wettability, frictional properties, adhesion and surface chemistry. The nanocoating technique using reactive fluorinated polymer nanosheets and surface modification of hard disks and microchannels are also reviewed. Copyright © 2010 Society of Chemical Industry
A micropump driven by a polypyrrole‐based conducting polymer soft actuator
Authors: Naka Yoshitaka , Fuchiwaki Masaki , Tanaka Kazuhiro
Published: February 2010

Micropumps are essential components of microfluidic systems and biosensing systems. In particular, micropumps used for micro total analysis systems transport fluids at a microflow rate with high precision. Moreover, they are also needed to transport high‐viscosity fluids in cases where there are various types of drugs to be transported. Therefore, in order to realize a micropump that can transport solutions at a microflow rate, it is necessary to develop a high‐precision microdeformable actuator with a simple mechanism. The conducting polymer soft actuator reported here opens widely and closes completely as a result of electrochemical oxidation and reduction, respectively. The opening and closing movement of the soft actuator, inside which the cation‐driven layer is arranged, becomes large because the anion‐driven layer that is arranged outside is the predominant driver. We developed a micropump driven by a conducting polymer soft actuator that opens and closes. Although the developed micropump contains no valve, the micropump can transport fluids in one direction without backflow. The newly developed micropump driven by the conducting polymer soft actuator can transport fluids in one direction without backflow by the opening and closing of two soft actuators. Furthermore, a wider range of flow rates and greater maximum delivery heads are obtained using the newly developed micropump. The energy consumption rate of the micropump is markedly lower than those of conventional micropumps. Copyright © 2010 Society of Chemical Industry
Perspectives for new dielectric elastomers with improved electromechanical actuation performance: composites versus blends
Authors: Gallone Giuseppe , Galantini Fabia , Carpi Federico
Published: February 2010

This work presents a comparison between composites and blends as two alternative approaches for the development of new dielectric elastomers offering superior electromechanical properties at lower driving electric fields. Silicone‐ and polyurethane‐based dielectric elastomers were modified by either making particulate composites with high‐permittivity ceramic fillers or by blending with different polymeric phases. Both previous and new data are considered. Experimental observations are compared and discussed to assess which one of the aforementioned approaches might be more generally preferable. Due to a consequent worsening of the mechanical properties, pure composite architectures yielded only limited results. While with the blend approach both an increase of the dielectric permittivity and an unexpected reduction of the tensile elastic modulus were observed, leading to an overall increase of the electromechanical response. The blending approach permits one to obtain a new dielectric elastomer with improved electromechanical properties by simply combining two low‐dielectric‐constant polymers. This is the first time such a type of result has been reported. This study indicates that formulating all‐polymeric compounds may represent a very promising route for obtaining new dielectric elastomers with improved actuation performance. Copyright © 2010 Society of Chemical Industry
Nonlinear oscillation of a dielectric elastomer balloon
Authors: Zhu Jian , Cai Shengqiang , Suo Zhigang
Published: February 2010

Much of the existing literature on dielectric elastomers has focused on quasi‐static deformation. However, in some potential applications, the elastomer deforms at high frequencies and undergoes nonlinear oscillation. While nonlinear oscillation has been studied in many areas of science and engineering, we are unaware of any theoretical analysis of nonlinear oscillation of dielectric elastomers. This paper reports a theoretical study of the dynamic behavior of a dielectric elastomer balloon subject to a combination of pressure and voltage. When the pressure and voltage are static, the balloon may reach a state of equilibrium. We determine the stability of the state of equilibrium, and calculate the natural frequency of the small‐amplitude oscillation around the state of equilibrium. We focus on the parametric responses of the dielectric elastomer balloon. When the voltage is sinusoidal, the balloon resonates at multiple frequencies of excitation, giving rise to superharmonic, harmonic and subharmonic responses. When the frequency of excitation varies continuously, the oscillating amplitude of the balloon may jump, exhibiting hysteresis. Copyright © 2010 Society of Chemical Industry
Conformational energy from the oxidation kinetics of poly(3,4‐ethylenedioxythiophene) films
Authors: Otero Toribio F , Caballero Romero María
Published: February 2010

Electroactive poly(3,4‐ethylenedioxythiophene) perchlorate films shrink and pack by electrochemical reduction at high cathodic potentials. Any subsequent oxidation by potential step gives chronoamperograms showing a maximum. At the maxima the electroactive oxidation reaction occurs under chemical kinetic control. Rate coefficients and activation energies for the reaction can be obtained by applying chemical kinetics methodology. Both parameters present two ranges as a function of the cathodic potential of pre‐polarization that defines the initial oxidation state of conformational packing. The activation energy includes two components: the constant chemical activation energy and the conformational packing energy that increases linearly after reduction–shrinking–packing at rising cathodic overpotentials. This methodology opens a way to quantify the conformational energy of polymer chains: the energy used by molecular motors constituting the actuator films of conducting polymers. Copyright © 2010 Society of Chemical Industry
Electro‐active polymer actuators employing sulfonated poly(styrene‐ran‐ethylene) as ionic membranes
Authors: Wang Xuan‐Lun , Oh Il‐Kwon , Cheng Tai‐Hong
Published: February 2010

We first investigated sulfonated poly(styrene‐ran‐ethylene) (SPSE) as a novel ionic membrane for electro‐active polymer actuators. SPSE is a newly developed hydrocarbon ionomer synthesized through a post‐sulfonation procedure. The thermal, electrical, morphological and mechanical properties, as well as ion‐exchange capacity, proton conductivity and water uptake were investigated and compared to a Nafion® counterpart. Tensile tests revealed that the SPSE membrane has good mechanical stiffness and strength while immersed in water. SPSE copolymers have good thermal stability and are easily processed for the production of membranes. In this study, SPSE actuators were fabricated according to traditional ionomeric polymer–metal composite preparation procedures and tailored to moderate dimensions. The bending responses of the SPSE actuators under both AC and DC electric excitations were investigated. The voltage–current behaviors of the actuators under AC excitation were also measured. The results showed that the SPSE actuators have good electromechanical performance with regard to tip displacement. Copyright © 2010 Society of Chemical Industry
Different deformation mechanisms of two modified‐polystyrene bimodal systems
Authors: Dai RuJun , Gao GuangHui , Zhang HuiXuan
Published: February 2010

It is well known that the dominant toughening mechanism of rubber‐modified polystyrene is multiple crazing. Some researchers have investigated polystyrene that can be modified by rubbers with dual particle sizes, leading to better mechanical properties. That is, the way to absorb energy during the deformation process is crazing and cavitation induced by rubber particles. Two types of polybutadiene‐graft‐polystyrene (PB‐g‐PS) rubber modifiers which have core‐shell structures were synthesized via an emulsion graft polymerization using redox and oil‐soluble initiators, respectively. To balance the yield strength, general‐purpose polystyrene was blended with the PB‐g‐PS modifiers, as well as commercial high‐impact polystyrene. Blends were defined as R‐bimodal and O‐bimodal corresponding to dispersed PB‐g‐PS particles formed using the redox and oil‐soluble initiators, respectively. The impact strength of R‐bimodal was improved significantly by altering the ratio of core to shell. However, little change of impact strength was observed for O‐bimodal. Transmission electron microscopy images of fracture surfaces indicated that the deformation mechanism of R‐bimodal is shear‐yielding induced by multi‐crazing. Moreover, PB‐g‐PS particles dispersed in O‐bimodal can form a ‘cluster’ structure, leading to crazing to absorb energy. Scanning electron microscopy images also showed obvious distinctness between the R‐bimodal and O‐bimodal systems due to different deformation mechanisms. Copyright © 2010 Society of Chemical Industry
The experience of bullying among secondary school students
Authors: Athanasiades Christina , Deliyanni‐Kouimtzis Vassiliki
Published: February 2010

The present study was designed to qualitatively investigate secondary students' interpretations and experiences of bullying (and victimization) in Greek schools, with a focus on gender similarities and differences. Overall, 95 students (50 boys and 45 girls), 15 or 16 years old, participated in focus group interviews that were homogeneous in terms of grade and gender. Data analysis, using the interpretative phenomenological approach, showed that different interpretations and meanings of bullying between genders have important consequences on actual behavior. Furthermore, students do not reveal bullying and victimization to either parents or teachers, who are described as indifferent and ineffective. Results are indicative of a school culture that is conducive to bullying behaviors and have important implications for antibullying interventions. © 2010 Wiley Periodicals, Inc.
Parent involvement and views of school success: The role of parents' Latino and White American cultural orientations
Authors: Ryan Carey S. , Casas Juan F. , Kelly‐Vance Lisa , Ryalls Brigette O. , Nero Collette
Published: February 2010

We examined ethnicity and cultural orientation as predictors of parents' views of and involvement in children's education, using data gathered from the Latino (n = 74) and non‐Latino (17 White and 13 ethnic minority) parents of children in an elementary school's dual‐language program. Parents completed a questionnaire that assessed Latino and White American cultural orientations, importance of children's academic and social success, and self‐ and significant other involvement in children's education. Results indicated that Latino (and other ethnic minority) parents valued academic and social success equally and more strongly than did Whites and that Whites valued social success more strongly than academic success. Latinos also reported greater involvement of significant others. These differences were largely accounted for by cultural orientations. Educational practices that take into account differences in cultural orientations and the involvement of significant others thus seem more likely to improve academic outcomes than do efforts intended to promote the valuing of education. © 2010 Wiley Periodicals, Inc.
Verification of protein disulfide bond arrangement by in‐gel tryptic digestion under entirely neutral pH conditions
Authors: Saito Kazuki , Yasuo Itsuki , Uchimura Hiromasa , Koide‐Yoshida Shizuyo , Mizuguchi Takaaki , Kiso Yoshiaki
Published: February 2010

To develop a concise proteomic procedure to verify the protein disulfide bond arrangement, non‐reductive trypsin digestion of neuregulin 1‐β1 (176–246), a model disulfide‐containing protein, was assessed by a proteolytic 18O‐labeling analysis. As a result, the commonly used in‐gel tryptic digestion method has been improved for use entirely under neutral pH conditions. With this procedure, the disulfide arrangement of proteins could represent a clinical index candidate in pathological proteomic studies.
Pathway analysis of dilated cardiomyopathy using global proteomic profiling and enrichment maps
Authors: Isserlin Ruth , Merico Daniele , Alikhani‐Koupaei Rasoul , Gramolini Anthony , Bader Gary D. , Emili Andrew
Published: February 2010

Global protein expression profiling can potentially uncover perturbations associated with common forms of heart disease. We have used shotgun MS/MS to monitor the state of biological systems in cardiac tissue correlating with disease onset, cardiac insufficiency and progression to heart failure in a time‐course mouse model of dilated cardiomyopathy. However, interpreting the functional significance of the hundreds of differentially expressed proteins has been challenging. Here, we utilize improved enrichment statistical methods and an extensive collection of functionally related gene sets, gaining a more comprehensive understanding of the progressive alterations associated with functional decline in dilated cardiomyopathy. We visualize the enrichment results as an Enrichment Map, where significant gene sets are grouped based on annotation similarity. This approach vastly simplifies the interpretation of the large number of enriched gene sets found. For pathways of specific interest, such as Apoptosis and the MAPK (mitogen‐activated protein kinase) cascade, we performed a more detailed analysis of the underlying signaling network, including experimental validation of expression patterns.
Protein–protein‐interactions in a multiplexed, miniaturized format a functional analysis of Rho GTPase activation and inhibition
Authors: Schmohl Michael , Rimmele Stefanie , Pötz Oliver , Kloog Yoel , Gierschik Peter , Joos Thomas O. , Schneiderhan‐Marra Nicole
Published: February 2010

A miniaturized, bead‐based protein–protein‐interaction assay was developed to study the interaction of Rho GTPases with regulatory proteins. The setup, which uses only minute amounts of sample, was used to analyze small molecules that inhibit the interaction between Rho GTPases and RhoGDIα. Prenylcysteine analogues and the replacement of GDP by non‐hydrolysable GTP analogues prevented the formation of Rho GTPase‐RhoGDIα complexes in a concentration‐dependent manner.
Proteomics of drug resistance in Candida glabrata biofilms
Authors: Seneviratne C. Jayampath , Wang Yu , Jin Lijian , Abiko Y. , Samaranayake Lakshman P.
Published: February 2010

Candida glabrata is a fungal pathogen that causes a variety of mucosal and systemic infections among compromised patient populations with higher mortality rates. Previous studies have shown that biofilm mode of the growth of the fungus is highly resistant to antifungal agents compared with the free‐floating or planktonic mode of growth. Therefore, in the present study, we used 2‐D DIGE to evaluate the differential proteomic profiles of C. glabrata under planktonic and biofilm modes of growth. Candida glabrata biofilms were developed on polystyrene surfaces and age‐matched planktonic cultures were obtained in parallel. Initially, biofilm architecture, viability, and antifungal susceptibility were evaluated. Differentially expressed proteins more than 1.5‐fold in DIGE analysis were subjected to MS/MS. The transcriptomic regulation of these biomarkers was evaluated by quantitative real‐time PCR. Candida glabrata biofilms were highly resistant to the antifungals and biocides compared with the planktonic mode of growth. Candida glabrata biofilm proteome when compared with its planktonic proteome showed upregulation of stress response proteins, while glycolysis enzymes were downregulated. Similar trend could be observed at transcriptomic level. In conclusion, C. glabrata biofilms possess higher amount of stress response proteins, which may potentially contribute to the higher antifungal resistance seen in C. glabrata biofilms.
Honokiol inhibits HepG2 migration via down‐regulation of IQGAP1 expression discovered by a quantitative pharmaceutical proteomic analysis
Authors: Liang Shufang , Fu Afu , Zhang Qiang , Tang Minghai , Zhou Jin , Wei Yuquan , Chen Lijuan
Published: February 2010

Honokiol (HNK), a natural small molecular product, inhibited proliferation of HepG2 cells and exhibited anti‐tumor activity in nude mice. In this article, we applied a novel sensitive stable isotope labeling with amino acids in cell culture‐based quantitative proteomic method and a model of nude mice to investigate the correlation between HNK and the hotspot migration molecule Ras GTPase‐activating‐like protein (IQGAP1). The quantitative proteomic analysis showed that IQGAP1 was 0.53‐fold down‐regulated under 10 μg/mL HNK exposure for 24 h on HepG2 cells. Migration ability of HepG2 cells under HNK treatment was correlated with its expression level of IQGAP1. In addition, the biochemical validation on HepG2 cells and the tumor xenograft model further demonstrated that HNK decreased the expression level of IQGAP1 and its upstream proteins Cdc42/Rac1. These data supported that HNK can modulate cell adhesion and cell migration by acting on Cdc42/Rac1 signaling via IQGAP1 interactions with its upstream Cdc42/Rac1 proteins, which is a new molecular mechanism of HNK to exert its anti‐tumor activity.
jmzML, an open‐source Java API for mzML, the PSI standard for MS data
Authors: Côté Richard G. , Reisinger Florian , Martens Lennart
Published: February 2010

We here present jmzML, a Java API for the Proteomics Standards Initiative mzML data standard. Based on the Java Architecture for XML Binding and XPath‐based XML indexer random‐access XML parser, jmzML can handle arbitrarily large files in minimal memory, allowing easy and efficient processing of mzML files using the Java programming language. jmzML also automatically resolves internal XML references on‐the‐fly. The library (which includes a viewer) can be downloaded from http://jmzml.googlecode.com.
Laser‐induced liquid bead ion desorption‐MS of protein complexes from blue‐native gels, a sensitive top‐down proteomic approach
Authors: Sokolova Lucie , Wittig Ilka , Barth Hans‐Dieter , Schägger Hermann , Brutschy Bernhard , Brandt Ulrich
Published: February 2010

We have developed an experimental approach that combines two powerful methods for proteomic analysis of large membrane protein complexes: blue native electrophoresis (BNE or BN‐PAGE) and laser‐induced liquid bead ion desorption (LILBID) MS. Protein complexes were separated by BNE and eluted from the gel. The masses of the constituents of the multiprotein complexes were obtained by LILBID MS, a detergent‐tolerant method that is especially suitable for the characterisation of membrane proteins. High sensitivity and small sample volumes required for LILBID MS resulted in low demands on sample quantity. Eluate from a single band allowed assessing the mass of an entire multiprotein complex and its subunits. The method was validated with mitochondrial NADH:ubiquinone reductase from Yarrowia lipolytica. For this complex of 947 kDa, typically 30 μg or 32 pmol were sufficient to obtain spectra from which the subunit composition could be analysed. The resolution of this electrophoretic small‐scale approach to the purification of native complexes was improved markedly by further separation on a second dimension of BNE. Starting from a subcellular fraction obtained by differential centrifugation, this allowed the purification and analysis of the constituents of a large multiprotein complex in a single LILBID spectrum.
Proteomic identification of interactions between histones and plasma proteins: Implications for cytoprotection
Authors: Pemberton Alan D. , Brown Jeremy K. , Inglis Neil F.
Published: February 2010

Extracellular histones released from cells during acute inflammation contribute to organ failure and death in a mouse model of sepsis, and histones are known to exert in vitro cytotoxicity in the absence of serum. Since addition of histones to serum and plasma is known to induce protein aggregation, we reasoned that plasma proteins may afford protection from cytotoxicity. We found that MODE‐K mouse small intestinal epithelial cells were protected from histone‐induced toxicity in the presence of 10% FCS. Therefore, the main aim of this study was to identify histone‐interacting plasma proteins that might be involved in cytoprotection. The precipitate formed following addition of calf thymus histones to human EDTA plasma was characterised by shotgun proteomics, identifying a total of 36 protein subunits, including complement components, coagulation factors, protease inhibitors and apolipoproteins. The highly sulphated glycosaminoglycan heparin inhibited histone‐induced plasma protein aggregation. Moreover, histones bound to heparin agarose were capable of pulling down plasma proteins from solution, indicating their effective cross‐linking properties. It was particularly notable that inter‐α‐trypsin inhibitor was prominent among the histone‐precipitated proteins, since it contains a chondroitin sulphate glycan chain, and suggests a potential role for this protein in histone sequestration during acute inflammation in vivo.
Single‐Chamber Deposition of Multilayer Barriers by Plasma Enhanced and Initiated Chemical Vapor Deposition of Organosilicones
Authors: Coclite Anna Maria , Ozaydin‐Ince Gozde , Palumbo Fabio , Milella Antonella , Gleason Karen K.
Published: February 2010

A novel technique, combining the plasma assisted deposition of SiOx‐like coatings with the initiated chemical vapor deposition (iCVD) of organosilicon films in a single‐chamber process, was investigated for the production of multistack barriers against the water vapor permeation. Hexavinyldisiloxane (HVDSO) was used as the film precursor for both kinds of polymerization. iCVD of HVDSO resulted in highly crosslinked and adherent carbon‐rich polymer which reduced the substrate roughness of the substrate, thus acting as a primer for the deposition of the denser C‐depleted uplayer. The plasma ion bombardment of the C‐rich underlayer produced a graded interphase which enhanced the adhesion between the layers and of the multilayer stack to the polymer substrate. The C‐rich interlayers effectively decoupled the defects of the C‐depleted layers, indeed a barrier improvement factor of 100 over the single C‐depleted barrier layer was obtained with a hexalayer structure.

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Reaction Chemistry of 1,4‐Benzopyrone Derivates in Non‐Equilibrium Low‐Temperature Plasmas
Authors: Grzegorzewski Franziska , Rohn Sascha , Quade Antje , Schröder Karsten , Ehlbeck Jörg , Schlüter Oliver , Kroh Lothar W.
Published: February 2010

1,4‐Benzopyrone derivates are exposed to different cold gas discharges. Reactions are carried out using different feed gases (argon, oxygen) and excitation sources (radio frequency, microwave) at both atmospheric and low pressure. A structure‐dependent degradation upon plasma‐chemical reactions can be observed. From contact‐angle measurements a strong surface oxidation is suggested. Independent of the source used plasma treatment leads to a significant increase in oxygen content of the samples through newly introduced carbonyl‐ and carboxyl‐functions. This is in agreement to results showing that during thermal food processes oxidative species lead to the formation of characteristic low‐molecular weight degradation products.

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A study of high‐dose oral silybin‐phytosome followed by prostatectomy in patients with localized prostate cancer
Authors: Flaig Thomas W. , Glodé Michael , Gustafson Daniel , van Bokhoven Adrie , Tao Yuzhen , Wilson Shandra , Su Lih‐Jen , Li Yuan , Harrison Gail , Agarwal Rajesh , Crawford E. David , Lucia M. Scott , Pollak Michael
Published: February 2010

BACKGROUND

Silibinin is a polyphenolic flavonolignan derived from milk thistle (Silybum marianium) with anti‐oxidant properties. The purpose of the current trial was to determine the tissue and blood effects of high‐dose silybin‐phytosome in prostate cancer patients.

METHODS

Subjects with localized prostate cancer planning for a prostatectomy were eligible to enroll. Six patients received 13 g of silybin‐phytosome daily with six additional participants serving as control subjects.

RESULTS

Patients in the treatment arm received silybin‐phytosome for 14–31 days (mean was 20 days) prior to surgery. Silibinin blood levels were measured 1 hr after the first silybin‐phytosome dose with a mean value of 19.7 µM. Trough silibinin levels were assessed at the end of the trial with an average concentration of 1.2 µM. In contrast to the high peak levels of silibinin observed in blood, the highest silibinin level observed in the harvested prostate tissue was 496.6 pmol/g. There were no significant differences noted in baseline and post‐treatment blood levels of IGF‐I and IGFBP‐3. One of the treated patients developed a grade 4 post‐operative thromboembolic event. The other observed toxicities in the treatment group were mild: four subjects had diarrhea and one had asymptomatic grade 2 hyperbilirubinemia which was transient.

CONCLUSIONS

High‐dose oral silybin‐phytosome achieves high blood concentrations transiently, but low levels of silibinin are seen in prostate tissue. Silibinin's lack of tissue penetration may be explained by its short half‐life, the brief duration of therapy in this study or an active process removing silibinin from the prostate. Prostate 70: 848–855, 2010. © 2010 Wiley‐Liss, Inc.
Fibronectin confers survival against chemotherapeutic agents but not against radiotherapy in DU145 prostate cancer cells: Involvement of the insulin like growth factor‐1 receptor
Authors: Thomas Francis , Holly Jeff M.P. , Persad Raj , Bahl Amit , Perks Claire M.
Published: February 2010

BACKGROUND

Tumor growth is influenced by an increase in cell proliferation and a reduction in apoptosis; both of which are affected by alterations in extracellular matrix (ECM). Our aim was to assess if the susceptibility of prostate cancer cells to apoptosis induced by either chemotherapeutics or radiotherapy was altered by changes in the ECM.

METHODS

Prostate cancer cell lines LNCaP and DU145 (androgen independent) cells were treated with chemotherapeutics (ceramide and docetaxel) or radiotherapy in the presence or absence of fibronectin, laminin, or vitronectin. Cell death was assessed using Trypan blue cell counting and apoptosis was confirmed by measuring PARP cleavage by Western immunoblotting (WIB). To identify a mechanism of action, changes in the abundance (WIB) or association (immunoprecipitation followed by WIB) of key proteins was also assessed.

RESULTS

We found that fibronectin, but not laminin or vitronectin activated a survival pathway that protected DU145 but not LNCaP prostate cancer cells against ceramide and docetaxel‐induced apoptosis but not that induced by radiotherapy. This survival effect involved the insulin‐like growth factor (IGF‐I) and β1 integrin receptors and was associated with an increase in the recruitment of the β1 integrin to a complex containing the IGF‐IR and protein receptor for activated C kinase (RACK‐1) and an increase in the abundance of a MAPK‐phosphatase‐1 (MKP‐1).

CONCLUSIONS

Changes in the ECM associated with disease progression may contribute to resistance to chemotherapeutic drugs but not to radiation therapy. The susceptibility to chemotherapy may be improved by targeting either the IGF‐I or β1 integrin receptors. Prostate 70: 856–865, 2010. © 2010 Wiley‐Liss, Inc.
Regulation of androgen receptor transactivity and mTOR–S6 kinase pathway by Rheb in prostate cancer cell proliferation
Authors: Kobayashi Takashi , Shimizu Yosuke , Terada Naoki , Yamasaki Toshinari , Nakamura Eijiro , Toda Yoshinobu , Nishiyama Hiroyuki , Kamoto Toshiyuki , Ogawa Osamu , Inoue Takahiro
Published: February 2010

BACKGROUND

Ras homolog‐enriched in brain (Rheb), a small GTP‐binding protein, is associated with prostate carcinogenesis through activating mammalian target of rapamycin (mTOR) signaling pathway. This study aimed to elucidate whether Rheb promotes proliferation of prostate cancer cells and can act as a potent therapeutic target in prostate cancer.

METHODS

Prostate cancer cell lines and human prostatic tissues were examined for the expression of Rheb. The effects of forced expression or knockdown of Rheb on cell proliferation were also examined. Semi‐quantitative and quantitative RT‐PCR were performed to evaluate mRNA expression. Western blotting was used to examine protein expression. Cell count and WST‐1 assay were used to measure cell proliferation. Fluorescence‐activated cell sorting was used to assess the cell cycle.

RESULTS

Rheb mRNA and protein expression was higher in more aggressive, androgen‐independent prostate cancer cell lines PC3, DU145, and C4‐2, compared with the less aggressive LNCaP. Rheb expression was higher in cancer tissues than in benign prostatic epithelia. Forced expression of Rheb in LNCaP cells accelerated proliferation without enhancing androgen receptor transactivity. Attenuation of Rheb expression or treatment with the mTOR inhibitor rapamycin decreased proliferation of PC3 and DU145 cells, with a decrease in the activated form of p70S6 kinase, one of the main targets of mTOR.

CONCLUSIONS

Rheb potentiates proliferation of prostate cancer cells and inhibition of Rheb or mTOR can lead to suppressed proliferation of aggressive prostate cancer cell lines in vitro. Rheb and the mTOR pathway are therefore probable targets for suppressing prostate cancer. Prostate 70: 866–874, 2010. © 2010 Wiley‐Liss, Inc.
Evaluation of the frequency of putative prostate cancer stem cells in primary and metastatic prostate cancer
Authors: Eaton Colby L. , Colombel Marc , van der Pluijm Gabri , Cecchini Marco , Wetterwald Antoinette , Lippitt Jenny , Rehman Ishtiaq , Hamdy Freddie , Thalman George
Published: February 2010

BACKGROUND

Tumour cells with a stem cell‐like phenotype have recently been identified in prostate tumors and it has been suggested that this population may be responsible for the diversity of cell types within tumors and also for the initiation of metastases. These cells carry a number of defined markers: they are cd133 and cd44+ve and express high levels of α2β1 integrin. In this study we have, for the first time, assessed matched primary and bone marrow biopsies from prostate cancer patients for the distribution of cells carrying these and a number of other putative stem cell markers.

METHODS

Eleven matched (primary and bone metastasis) specimens from prostate cancer patients were assessed for the presence of cd133, cd44, α2β1 integrin, CXCR4, c‐met, α6 integrin, and nestin using immunohistochemistry and stain intensity and distribution scored.

RESULTS

In the bone metastases, tumor cells staining positively for cd133 were detected at low frequency in ∼50% of samples. Staining for nestin was confined to endothelium. Positive staining of tumor cells for the other antigens was present at variable frequency in >70% of metastases with the exception of CXCR4 which was absent from all but 2 specimens. Where positive staining of tumor cells was present in the metastasis, cells staining for each antigen were present in the matched primary with the exception of cd44 which was absent in all but 2/11 matched primary tissues.

CONCLUSIONS

In established metastases no single or combination of marker expression profiles identify the established metastatic phenotype, although cd44 expression was shown to be more frequent in metastases that in primary cancers. Prostate 70: 875–882, 2010. © 2010 Wiley‐Liss, Inc.
Genetic alterations and changes in expression of histone demethylases in prostate cancer
Authors: Suikki Hanna E. , Kujala Paula M. , Tammela Teuvo L.J. , van Weerden Wytske M. , Vessella Robert L. , Visakorpi Tapio
Published: February 2010

BACKGROUND

Histone demethylases LSD1, JHDM2A, and GASC1 have been suggested to function as androgen receptor co‐activators, and to be involved in prostate cancer (PC) progression. We aim to identify genetic alterations and changes in expression of these genes in PC.

METHODS

PC cell lines, xenografts as well as clinical specimens were screened for mutations using denaturating high‐performance liquid chromatography and sequencing, and for expression alterations by using quantitative RT‐PCR and immunohistochemistry.

RESULTS

Only known single nucleotide polymorphisms, but no mutations, were found in these genes. JHDMA2 mRNA expression was slightly increased (P < 0.05) in PC compared with benign prostate hyperplasia (BPH), whereas the expression of GASC1 was slightly higher (P < 0.05) in castration‐resistant PC (CRPC) compared with untreated PC or BPH. The mRNA expression of LSD1 was not altered in PC. The expression of LSD1 protein was somewhat, although not statistically significantly (P = 0.0521) lower in CRPC compared with untreated PC. In prostatectomy specimens, the level of LSD1 protein expression was associated with low pT‐stage (P = 0.0402), but not with Gleason score or progression‐free survival.

CONCLUSIONS

As no genetic alterations and only very modest expression changes were found, it is unlikely that LSD1, JHDM2A, or GASC1 play a major role in the progression of PC. Prostate 70: 889–898, 2010. © 2010 Wiley‐Liss, Inc.
Binding of calcium is sensed structurally and dynamically throughout the second calcium‐binding domain of the sodium/calcium exchanger
Authors: Breukels Vincent , Vuister Geerten W.
Published: February 2010

We report the effects of Ca2+ binding on the backbone relaxation rates and chemical shifts of the AD and BD splice variants of the second Ca2+‐binding domain (CBD2) of the sodium–calcium exchanger. Analysis of the Ca2+‐induced chemical shifts perturbations yields similar KD values of 16–24 μM for the two CBD2‐AD Ca2+‐binding sites, and significant effects are observed up to 20 Å away. To quantify the Ca2+‐induced chemical shift changes, we performed a comparative analysis of eight Ca2+‐binding proteins that revealed large differences between different protein folds. The CBD2 15N relaxation data show the CBD2‐AD Ca2+ coordinating loops to be more rigid in the Ca2+‐bound state as well as to affect the FG‐loop located at the opposite site of the domain. The equivalent loops of the CBD2‐BD splice variant do not bind Ca2+ and are much more dynamic relative to both the Ca2+‐bound and apo forms of CBD2‐AD. A more structured FG‐loop in CBD2‐BD is suggested by increased S2 order parameter values relative to both forms of CBD2‐AD. The chemical shift and relaxation data together indicate that, in spite of the small structural changes, the Ca2+‐binding event is felt throughout the molecule. The data suggest that the FG‐loop plays an important role in connecting the Ca2+‐binding event with the other cytosolic domains of the NCX, in line with in vivo and in vitro biochemical data as well as modeling results that connect the CBD2 FG‐loop with the first Ca2+‐binding domain of NCX. Proteins 2010. © 2010 Wiley‐Liss, Inc.
Contact prediction for beta and alpha‐beta proteins using integer linear optimization and its impact on the first principles 3D structure prediction method ASTRO‐FOLD
Authors: Rajgaria R. , Wei Y. , Floudas C. A.
Published: February 2010

An integer linear optimization model is presented to predict residue contacts in β, α + β, and α/β proteins. The total energy of a protein is expressed as sum of aCαCαdistance dependent contact energy contribution and a hydrophobic contribution. The model selects contact that assign lowest energy to the protein structure as satisfying a set of constraints that are included to enforce certain physically observed topological information. A new method based on hydrophobicity is proposed to find the β‐sheet alignments. These β‐sheet alignments are used as constraints for contacts between residues of β‐sheets. This model was tested on three independent protein test sets and CASP8 test proteins consisting of β, α + β, α/β proteins and it was found to perform very well. The average accuracy of the predictions (separated by at least six residues) was ∼61%. The average true positive and false positive distances were also calculated for each of the test sets and they are 7.58 Å and 15.88 Å, respectively. Residue contact prediction can be directly used to facilitate the protein tertiary structure prediction. This proposed residue contact prediction model is incorporated into the first principles protein tertiary structure prediction approach, ASTRO‐FOLD. The effectiveness of the contact prediction model was further demonstrated by the improvement in the quality of the protein structure ensemble generated using the predicted residue contacts for a test set of 10 proteins. Proteins 2010. © 2010 Wiley‐Liss, Inc.
Structural rationale for the short branched substrate specificity of the glycogen debranching enzyme GlgX
Authors: Song Hyung‐Nam , Jung Tae‐Yang , Park Jong‐Tae , Park Byung‐Chul , Myung Pyung Keun , Boos Winfried , Woo Eui‐Jeon , Park Kwan‐Hwa
Published: February 2010

Glycogen serves as major energy storage in most living organisms. GlgX, with its gene in the glycogen degradation operon, functions in glycogen catabolism by selectively catalyzing the debranching of polysaccharide outer chains in bacterial glycosynthesis. GlgX hydrolyzes α‐1,6‐glycosidic linkages of phosphorylase‐limit dextrin containing only three or four glucose subunits produced by glycogen phosphorylase. To understand its mechanism and unique substrate specificity toward short branched α‐polyglucans, we determined the structure of GlgX from Escherichia Coli K12 at 2.25 Å resolution. The structure reveals a monomer consisting of three major domains with high structural similarity to the subunit of TreX, the oligomeric bifunctional glycogen debranching enzyme (GDE) from Sulfolobus. In the overlapping substrate binding groove, conserved residues Leu270, Asp271, and Pro208 block the cleft, yielding a shorter narrow GlgX cleft compared to that of TreX. Residues 207–213 form a unique helical conformation that is observed in both GlgX and TreX, possibly distinguishing GDEs from isoamylases and pullulanases. The structural feature observed at the substrate binding groove provides a molecular explanation for the unique substrate specificity of GlgX for G4 phosphorylase‐limit dextrin and the discriminative activity of TreX and GlgX toward substrates of varying lengths. Proteins 2010. © 2010 Wiley‐Liss, Inc.
Dewetting transitions in protein cavities
Authors: Young Tom , Hua Lan , Huang Xuhui , Abel Robert , Friesner Richard , Berne B. J.
Published: February 2010

In a previous analysis of the solvation of protein active sites, a drying transition was observed in the narrow hydrophobic binding cavity of Cox‐2. With the use of a crude metric that often seems able to discriminate those protein cavities that dry from those that do not, we made an extensive search of the PDB, and identified five other proteins that, in molecular dynamics simulations, undergo drying transitions in their active sites. Because such cavities need not desolvate before binding hydrophobic ligands they often exhibit very large binding affinities. This article gives evidence that drying in protein cavities is not unique to Cox‐2. Proteins 2010. © 2010 Wiley‐Liss, Inc.