bluuebunny/crossref_metadata_2025 huggingface dataset:
Environmental parking charging policies: A case study of Winchester
Authors: Wall Graham
Published: June 2011

This article reports the results of a package of environmentally-linked parking charging policies designed to reduce the environmental impact of the parking fleet in Winchester. These measures were implemented as part of the EU-sponsored CIVITAS MIRACLES project which ran from 2002 to 2006. The measures included a variable tariff for annual parking permit holders at several long stay car parks in the city centre, offering a discount of 75 percent or 50 percent for those vehicles in the two lowest CO2 tax bands, with free annual permits offered to owners of electric or hybrid vehicles. In addition, a differential parking charging policy was implemented to discourage long stay parking in the city centre and encourage the use of off-centre and peripheral car parks such as the Park and Ride. This article includes the results of the scheme, a review of other existing schemes and the conclusions and recommendations drawn from the evidence.
A novel method of determining residual stress distributions in plates using the incremental slitting technique
Authors: Roberts S J , McCartney L N , Broughton W R
Published: June 2011

A method is described to determine residual stress distributions in homogeneous plates from strain measurements obtained using the incremental slitting method, which is modelled using a linear thermo-elastic analysis originally developed for anisotropic multiple-ply laminates having arrays of equally spaced ply cracks. A linear thermo-elastic analysis, for the special case where the plate is homogeneous and isotropic, is used within an inverse modelling problem to interpret measured strains, obtained by gradually slitting the plate and measuring strain changes on the external surface during bend deformation, in terms of distributions of residual stress. The model has been used to estimate the residual stress distributions within polypropylene-based samples, including samples with untreated and aminosilane-treated glass flakes. In the internal region of the plate, the residual stress distribution can be estimated reliably as results do not depend on the parameters used for the analysis. Results of predictions near the cut free surface show some variability, which is thought to be attributable to the insensitivity of strain measurements to the length of the slit. To overcome this problem it is recommended that a plate is slit from both free surfaces at different widely spaced locations. Comparisons of the calculated strains with the original smoothed experimental input data showed very close agreement, indicating that the linear thermo-elastic model is appropriate for these samples.
Fabrication of new composite abrasive for jet machining and application to scrap wafer regeneration
Authors: Ke J-H , Tsai F-C , Hung J-C , Yang T-Y , Yan B-H
Published: June 2011

This study develops a novel composite abrasive with better micro-machining properties for achieving improved surface roughness on hard brittle materials. The proposed composite abrasive is made of a thermoplastic polymer with polystyrene as the base material and has its surface thermally coated with SiC particles for abrasive jet machining (AJM). The optimal parameters for obtaining an even coating of abrasives on the polymer surface include coating temperature of 200°C and SiC particle size of #3000. Application of the new composite abrasive to AJM achieves enhancement in surface roughness. Being elastic and spherical, the composite abrasive can function as a buffer preventing direct impact on the workpiece, resulting in smaller, fewer, and shallower cracks. When applied to regeneration of scrap silicon wafers, the developed composite abrasive contributes to both improvement in surface roughness and shortening of the post-processing time required.
Media navel-gazing: not dead yet
Authors: Snoddy Raymond
Published: June 2011

Expectations of large profits drove the quality press to take up navel-gazing. The dream faded, but now media could be good news again
The transfer of V2: inversion and negation in German and Dutch learners of English
Authors: Rankin Tom
Published: June 2011

This article investigates the transfer of verb-second syntax (V2) from L1 German and Dutch into L2 English. A comparative learner corpus analysis between L1 German, Dutch and French, and native English writing reveals that the German and Dutch speakers produce distinct patterns of inversion in declarative clauses, indicating the transfer of V2. They produce non-target subject–auxiliary inversion and copula inversion. However, other reflexes of V2 in interrogatives or with sentential negation are not produced. This is analysed as evidence that the German and Dutch learners have mastered the syntax of English but transfer continues to occur at the level of discourse-pragmatics, where L1 preferences for topicalization structures continue to transfer. This is in line with predictions of the Interface Hypothesis in second language acquisition, which assumes that the interfaces of syntax with other modules of the grammar are more difficult to acquire and may continue to show L1 effects even after the narrow syntax has been mastered.
‘Gray, meticulous and patently documentary’
Authors: Dent Chris
Published: June 2011

Patents are seen as a key part of the modern economy and operate as a mode of regulation of technology and innovation. The histories of the system, to date, have not explored the role that patents have in the governance of our society. This article suggests that the historical methods of Michel Foucault are broad enough to undertake this task. The article, then, explores both the archaeological and the genealogical methods in order to assess the benefits, and limitations, that arise from the use of each of them in the context of a history of the laws, and practices, as they relate to patents of invention.
Reliability analysis of screwed connections in high-speed trains, considering fatigue, corrosion, and imperfect maintenance operations
Authors: Chateauneuf A , Cocheteux F , Deffarges F , Sourget F
Published: June 2011

The reliability of railway equipment is of central interest in the choice of design and maintenance strategies. In high-speed trains, known as TGV, the assembly of mechanical parts is performed by the use of prestressed screws. In order to ensure safe operation of the railway network, the French company SNCF has undertaken a reliability-based analysis of the life cycle of screwed connections. This study aims at establishing a consistent reliability-based methodology for the assessment of equipments subjected to fatigue, corrosion, and imperfect maintenance operations, in order to improve their inspection on the basis of cost and reliability balancing. This methodology incorporates the life cycle of the equipment, including the deterioration modes, the imperfect maintenance operations and the uncertainties in geometry, loading, and material properties. The results of the study show that despite the high resistance regarding the applied load, the connection is still very sensitive to parameter variations: friction, tightening and prestressing. Although the large safety margin in fatigue design, the robustness of screwed connections should be sought when operation and maintenance conditions are considered.
Probabilistic analysis of shear fatigue life of steel plate girders using a fracture mechanics approach
Authors: Anoop M B , Prabhu G , Balaji Rao K
Published: June 2011

Probabilistic analysis of shear fatigue life of steel plate girders of two railway bridges with welded connections, designed according to Indian Railway Standards, is presented in this paper. The fatigue life is determined using a fracture mechanics based procedure. Deterministic and probabilistic sensitivity analyses are carried out and, based on the results of these sensitivity analyses, the random variables having significant influence on fatigue life are identified. The probability distribution of fatigue life is determined using the Monte Carlo simulation technique and a relation for estimating the characteristic fatigue life is proposed.
Risk-based inspections enhanced with Bayesian networks
Authors: De Carlo F , Borgia O , Tucci M
Published: June 2011

The present paper aims to assess the usefulness of the integration of a qualitative risk-based inspection (RBI) procedure with the modelling of Bayesian belief networks (BBNs). In such a way, qualitative RBI, usually performed following standard procedures, might still be applied, even with the lack of some necessary data. Another benefit of the proposed method is the capability to include new factors in the evaluation, overtaking the rigid structure of the procedure. The qualitative RBI was chosen for its simple and clear structure, while the choice of the BBN formalism was made because of its flexibility and power to represent discontinuous variables. Bayesian formalism, moreover, allows a quick and easy extension of the model with additional variables. As a result, a BBN modelling a qualitative RBI was constructed. In the network, all of the RBI variables were included, as well as new external variables. An application for a chemical plant was tested in order to prove that BBNs can profitably model qualitative RBIs and can go beyond its rigid structure with the addition of new variables.
Long-term outcome and lineage-specific chimerism in 194 patients with Wiskott-Aldrich syndrome treated by hematopoietic cell transplantation in the period 1980-2009: an international collaborative study
Authors: Moratto Daniele , Giliani Silvia , Bonfim Carmem , Mazzolari Evelina , Fischer Alain , Ochs Hans D. , Cant Andrew J. , Thrasher Adrian J. , Cowan Morton J. , Albert Michael H. , Small Trudy , Pai Sung-Yun , Haddad Elie , Lisa Antonella , Hambleton Sophie , Slatter Mary , Cavazzana-Calvo Marina , Mahlaoui Nizar , Picard Capucine , Torgerson Troy R. , Burroughs Lauri , Koliski Adriana , Neto Jose Zanis , Porta Fulvio , Qasim Waseem , Veys Paul , Kavanau Kristina , Hönig Manfred , Schulz Ansgar , Friedrich Wilhelm , Notarangelo Luigi D.
Published: June 2011

In this retrospective collaborative study, we have analyzed long-term outcome and donor cell engraftment in 194 patients with Wiskott-Aldrich syndrome (WAS) who have been treated by hematopoietic cell transplantation (HCT) in the period 1980- 2009. Overall survival was 84.0% and was even higher (89.1% 5-year survival) for those who received HCT since the year 2000, reflecting recent improvement of outcome after transplantation from mismatched family donors and for patients who received HCT from an unrelated donor at older than 5 years. Patients who went to transplantation in better clinical conditions had a lower rate of post-HCT complications. Retrospective analysis of lineage-specific donor cell engraftment showed that stable full donor chimerism was attained by 72.3% of the patients who survived for at least 1 year after HCT. Mixed chimerism was associated with an increased risk of incomplete reconstitution of lymphocyte count and post-HCT autoimmunity, and myeloid donor cell chimerism < 50% was associated with persistent thrombocytopenia. These observations indicate continuous improvement of outcome after HCT for WAS and may have important implications for the development of novel protocols aiming to obtain full correction of the disease and reduce post-HCT complications.
Delivery of progenitors to the thymus limits T-lineage reconstitution after bone marrow transplantation
Authors: Zlotoff Daniel A. , Zhang Shirley L. , De Obaldia Maria Elena , Hess Paul R. , Todd Sarah P. , Logan Theodore D. , Bhandoola Avinash
Published: June 2011

T-cell production depends on the recruitment of hematopoietic progenitors into the thymus. T cells are among the last of the hematopoietic lineages to recover after bone marrow transplantation (BMT), but the reasons for this delay are not well understood. Under normal physiologic conditions, thymic settling is selective and either CCR7 or CCR9 is required for progenitor access into the thymus. The mechanisms of early thymic reconstitution after BMT, however, are unknown. Here we report that thymic settling is briefly CCR7/CCR9-independent after BMT but continues to rely on the selectin ligand PSGL-1. The CCR7/CCR9 independence is transient, and by 3 weeks after BMT these receptors are again strictly required. Despite the normalization of thymic settling signals, the rare bone marrow progenitors that can efficiently repopulate the thymus are poorly reconstituted for at least 4 weeks after BMT. Consistent with reduced progenitor input to the thymus, intrathymic progenitor niches remain unsaturated for at least 10 weeks after BMT. Finally, we show that thymic recovery is limited by the number of progenitors entering the thymus after BMT. Hence, T-lineage reconstitution after BMT is limited by progenitor supply to the thymus.
Distribution of Bim determines Mcl-1 dependence or codependence with Bcl-xL/Bcl-2 in Mcl-1–expressing myeloma cells
Authors: Morales Alejo A. , Kurtoglu Metin , Matulis Shannon M. , Liu Jiangxia , Siefker David , Gutman Delia M. , Kaufman Jonathan L. , Lee Kelvin P. , Lonial Sagar , Boise Lawrence H.
Published: June 2011

Dependence on Bcl-2 proteins is a common feature of cancer cells and provides a therapeutic opportunity. ABT-737 is an antagonist of antiapoptotic Bcl-2 proteins and therefore is a good predictor of Bcl-xL/Bcl-2 dependence. Surprisingly, analysis of Mcl-1–dependent multiple myeloma cell lines revealed codependence on Bcl-2/Bcl-xL in half the cells tested. Codependence is not predicted by the expression level of antiapoptotic proteins, rather through interactions with Bim. Consistent with these findings, acquired resistance to ABT-737 results in loss of codependence through redistribution of Bim to Mcl-1. Overall, these results suggest that complex interactions, and not simply expression patterns of Bcl-2 proteins, need to be investigated to understand Bcl-2 dependence and how to better use agents, such as ABT-737.
Translocation of signalling proteins to the plasma membrane revealed by a new bioluminescent procedure
Authors: Giorgi Carlotta , Romagnoli Anna , Agnoletto Chiara , Bergamelli Leda , Sorrentino Giovanni , Brini Marisa , Pozzan Tullio , Meldolesi Jacopo , Pinton Paolo , Rizzuto Rosario
Published: June 2011

Background

Activation by extracellular ligands of G protein-coupled (GPCRs) and tyrosine kinase receptors (RTKs), results in the generation of second messengers that in turn control specific cell functions. Further, modulation/amplification or inhibition of the initial signalling events, depend on the recruitment onto the plasma membrane of soluble protein effectors.

High throughput methodologies to monitor quantitatively second messenger production, have been developed over the last years and are largely used to screen chemical libraries for drug development. On the contrary, no such high throughput methods are yet available for the other aspect of GPCRs regulation, i.e. protein translocation to the plasma membrane, despite the enormous interest of this phenomenon for the modulation of receptor downstream functions. Indeed, to date, the experimental procedures available are either inadequate or complex and expensive.

Results

Here we describe the development of a novel conceptual approach to the study of cytosolic proteins translocation to the inner surface of the plasma membrane. The basis of the technique consists in: i) generating chimeras between the protein of interests and the calcium (Ca2+)-sensitive, luminescent photo-protein, aequorin and ii) taking advantage of the large Ca2+concentration [Ca2+] difference between bulk cytosolic and the sub-plasma membrane rim.

Conclusion

This approach, that keeps unaffected the translocation properties of the signalling protein, can in principle be applied to any protein that, upon activation, moves from the cytosol to the plasma membrane.

Thus, not only the modulation of GPCRs and RTKs can be investigated in this way, but that of all other proteins that can be recruited to the plasma membrane also independently of receptor activation.

Moreover, its automated version, which can provide information about the kinetics and concentration-dependence of the process, is also applicable to high throughput screening of drugs affecting the translocation process.
Heterologous expression of pathogen-specific genes ligA and ligB in the saprophyte Leptospira biflexaconfers enhanced adhesion to cultured cells and fibronectin
Authors: Figueira Cláudio Pereira , Croda Julio , Choy Henry A , Haake David A , Reis Mitermayer G , Ko Albert I , Picardeau Mathieu
Published: June 2011

Background

In comparison to other bacterial pathogens, our knowledge of the molecular basis of the pathogenesis of leptospirosis is extremely limited. An improved understanding of leptospiral pathogenetic mechanisms requires reliable tools for functional genetic analysis. Leptospiral immunoglobulin-like (Lig) proteins are surface proteins found in pathogenic Leptospira, but not in saprophytes. Here, we describe a system for heterologous expression of the Leptospira interrogans genes ligA and ligB in the saprophyte Leptospira biflexa serovar Patoc.

Results

The genes encoding LigA and LigB under the control of a constitutive spirochaetal promoter were inserted into the L. biflexa replicative plasmid. We were able to demonstrate expression and surface localization of LigA and LigB in L. biflexa. We found that the expression of the lig genes significantly enhanced the ability of transformed L. biflexa to adhere in vitro to extracellular matrix components and cultured cells, suggesting the involvement of Lig proteins in cell adhesion.

Conclusions

This work reports a complete description of the system we have developed for heterologous expression of pathogen-specific proteins in the saprophytic L. biflexa. We show that expression of LigA and LigB proteins from the pathogen confers a virulence-associated phenotype on L. biflexa, namely adhesion to eukaryotic cells and fibronectin in vitro. This study indicates that L. biflexa can serve as a surrogate host to characterize the role of key virulence factors of the causative agent of leptospirosis.
Learning alters theta amplitude, theta-gamma coupling and neuronal synchronization in inferotemporal cortex
Authors: Kendrick Keith M , Zhan Yang , Fischer Hanno , Nicol Alister U , Zhang Xuejuan , Feng Jianfeng
Published: June 2011

Background

How oscillatory brain rhythms alone, or in combination, influence cortical information processing to support learning has yet to be fully established. Local field potential and multi-unit neuronal activity recordings were made from 64-electrode arrays in the inferotemporal cortex of conscious sheep during and after visual discrimination learning of face or object pairs. A neural network model has been developed to simulate and aid functional interpretation of learning-evoked changes.

Results

Following learning the amplitude of theta (4-8 Hz), but not gamma (30-70 Hz) oscillations was increased, as was the ratio of theta to gamma. Over 75% of electrodes showed significant coupling between theta phase and gamma amplitude (theta-nested gamma). The strength of this coupling was also increased following learning and this was not simply a consequence of increased theta amplitude. Actual discrimination performance was significantly correlated with theta and theta-gamma coupling changes. Neuronal activity was phase-locked with theta but learning had no effect on firing rates or the magnitude or latencies of visual evoked potentials during stimuli. The neural network model developed showed that a combination of fast and slow inhibitory interneurons could generate theta-nested gamma. By increasing N-methyl-D-aspartate receptor sensitivity in the model similar changes were produced as in inferotemporal cortex after learning. The model showed that these changes could potentiate the firing of downstream neurons by a temporal desynchronization of excitatory neuron output without increasing the firing frequencies of the latter. This desynchronization effect was confirmed in IT neuronal activity following learning and its magnitude was correlated with discrimination performance.

Conclusions

Face discrimination learning produces significant increases in both theta amplitude and the strength of theta-gamma coupling in the inferotemporal cortex which are correlated with behavioral performance. A network model which can reproduce these changes suggests that a key function of such learning-evoked alterations in theta and theta-nested gamma activity may be increased temporal desynchronization in neuronal firing leading to optimal timing of inputs to downstream neural networks potentiating their responses. In this way learning can produce potentiation in neural networks simply through altering the temporal pattern of their inputs.
Using e-mail recruitment and an online questionnaire to establish effect size: A worked example
Authors: Kirkby Helen M , Wilson Sue , Calvert Melanie , Draper Heather
Published: June 2011

Background

Sample size calculations require effect size estimations. Sometimes, effect size estimations and standard deviation may not be readily available, particularly if efficacy is unknown because the intervention is new or developing, or the trial targets a new population. In such cases, one way to estimate the effect size is to gather expert opinion. This paper reports the use of a simple strategy to gather expert opinion to estimate a suitable effect size to use in a sample size calculation.

Methods

Researchers involved in the design and analysis of clinical trials were identified at the University of Birmingham and via the MRC Hubs for Trials Methodology Research. An email invited them to participate.

An online questionnaire was developed using the free online tool 'Survey Monkey©'. The questionnaire described an intervention, an electronic participant information sheet (e-PIS), which may increase recruitment rates to a trial. Respondents were asked how much they would need to see recruitment rates increased by, based on 90%. 70%, 50% and 30% baseline rates, (in a hypothetical study) before they would consider using an e-PIS in their research.

Analyses comprised simple descriptive statistics.

Results

The invitation to participate was sent to 122 people; 7 responded to say they were not involved in trial design and could not complete the questionnaire, 64 attempted it, 26 failed to complete it. Thirty-eight people completed the questionnaire and were included in the analysis (response rate 33%; 38/115). Of those who completed the questionnaire 44.7% (17/38) were at the academic grade of research fellow 26.3% (10/38) senior research fellow, and 28.9% (11/38) professor. Dependent upon the baseline recruitment rates presented in the questionnaire, participants wanted recruitment rate to increase from 6.9% to 28.9% before they would consider using the intervention.

Conclusions

This paper has shown that in situations where effect size estimations cannot be collected from previous research, opinions from researchers and trialists can be quickly and easily collected by conducting a simple study using email recruitment and an online questionnaire. The results collected from the survey were successfully used in sample size calculations for a PhD research study protocol.
Experiences of outreach workers in promoting smoking cessation to Bangladeshi and Pakistani men: longitudinal qualitative evaluation
Authors: Begh Rachna A , Aveyard Paul , Upton Penney , Bhopal Raj S , White Martin , Amos Amanda , Prescott Robin J , Bedi Raman , Barton Pelham M , Fletcher Monica , Gill Paramjit , Zaidi Qaim , Sheikh Aziz
Published: June 2011

Background

Despite having high smoking rates, there have been few tailored cessation programmes for male Bangladeshi and Pakistani smokers in the UK. We report on a qualitative evaluation of a community-based, outreach worker delivered, intervention that aimed to increase uptake of NHS smoking cessation services and tailor services to meet the needs of Bangladeshi and Pakistani men.

Methods

This was a longitudinal, qualitative study, nested within a phase II cluster randomised controlled trial of a complex intervention. We explored the perspectives and experiences of five outreach workers, two stop smoking service managers and a specialist stop smoking advisor. Data were collected through focus group discussions, weekly diaries, observations of management meetings, shadowing of outreach workers, and one-to-one interviews with outreach workers and their managers. Analysis was undertaken using a modified Framework approach.

Results

Outreach workers promoted cessation services by word of mouth on the streets, in health service premises, in local businesses and at a wide range of community events. They emphasised the reasons for cessation, especially health effects, financial implications, and the impact of smoking on the family. Many smokers agreed to be referred to cessation services, but few attended, this in part being explained by concerns about the relative inflexibility of existing service provision. Although outreach workers successfully expanded service reach, they faced the challenges of perceived lack of awareness of the health risks associated with smoking in older smokers and apathy in younger smokers. These were compounded by perceptions of "lip service" being given to their role by community organisations and tensions both amongst the outreach workers and with the wider management team.

Conclusions

Outreach workers expanded reach of the service through taking it to diverse locations of relevance to Pakistani and Bangladeshi communities. The optimum method of outreach to retain and treat Bangladeshi and Pakistani smokers effectively in cessation programmes needs further development.
Drug Checking: A prevention measure for a heterogeneous group with high consumption frequency and polydrug use - evaluation of zurich's drug checking services
Authors: Hungerbuehler Ines , Buecheli Alexander , Schaub Michael
Published: June 2011

Background

The increasing party culture in Zurich presents new challenges, especially regarding the consumption of alcohol and so-called party drugs. Streetwork, the youth advisory service of the city of Zurich, has provided onsite and stationary Drug Checking facilities since 2001 and 2006, respectively. Drug Checking always involves filling out an anonymous questionnaire, which allows the collection of important information about a largely unknown group of users and their consumption patterns.

Methods

The questionnaires assessed sociodemographic characteristics, consumption patterns, Drug Checking experiences, information behavior and social support. The collected data were statistically analyzed by the Research Institute for Public Health and Addiction (RIPHA).

Results

The majority of Drug Checking service patrons were male and between 20 and 35 years old. These patrons reported high lifetime prevalences and high consumption frequencies of legal and illegal substances, and they often reported polydrug use. Aside from tobacco and alcohol, the most consumed drugs during typical party nights were ecstasy, amphetamines, cannabis and cocaine. Party drug consumers using Drug Checking services form a heterogeneous group with respect to sociodemographic characteristics and consumption patterns. Users of the onsite Drug Checking facilities were significantly younger, were less experienced with drug testing, and reported more polydrug use than users of the stationary Drug Checking service.

Conclusions

Drug Checking combined with a consultation appears to be an important harm reduction and prevention measure that reaches a group of consumers with high consumption frequency and polydrug use. Because of the heterogeneity of the target group, different prevention measures must be offered and embedded in an overall local concept.
HCV genotype-specific correlation with serum markers: Higher predictability for genotype 4a
Authors: Ahmad Waqar , Ijaz Bushra , Javed Fouzia T , Kausar Humera , Sarwar Muhammad T , Gull Sana , Asad Sultan , Shahid Imran , Hassan Sajida
Published: June 2011

Background

Several factors have been proposed to assess the clinical outcome of HCV infection. The correlation of HCV genotypes to possible serum markers in clinical prediction is still controversial. The main objective of this study was to determine the existence of any correlation between HCV genotypes to viral load and different clinical serum markers.

Methods

We performed a prospective cross-sectional and observational study. About 3160 serum HCV RNA positive patients were chosen from 4020 randomly selected anti-HCV positive patients. Statistical analysis was performed using the SPSS 16 software package. ROC (receiver operating characteristics) curves were used to compare diagnostic values of serum markers to predict genotypes.

Results

The most prevalent genotype was 3a (73.9%) followed by 1a (10.7%), 4a (6.4%) and 3b (6.1%) in Pakistani population. No correlation was found between viral load and serum markers for genotype 3a in a large no. of sample (n = 2336). While significant correlation was observed between viral load and AST in genotype 3b, ALP with viral load and ALT for genotype 1a. Patients with genotype 4a showed a significant inverse correlation with viral load and Hb level and AST with ALP. For genotype 4a, AUC (area under the curve) of ALT, ALP, AST, bilirubin, Hb level and viral load was 0.790, 0.763, 0.454, 0.664, 0.458 and 0.872 respectively.

Conclusions

In conclusion, there was a significant variable response of HCV genotypes with serum markers. Severity of disease is independent of serum marker level in genotype 3a, while the liver damage in genotype 4a may associate with viral cytopathic effect as well as the immune-mediated process. An index using six serum markers may correctly predict genotype 4a in patients with ≥75% accuracy.
Efficient virus-induced gene silencing in apple, pear and Japanese pear using Apple latent spherical virus vectors
Authors: Sasaki Shintarou , Yamagishi Noriko , Yoshikawa Nobuyuki
Published: June 2011

Background

Virus-induced gene silencing (VIGS) is an effective technology for the analysis of gene functions in plants. Though there are many reports on virus vectors for VIGS in plants, no VIGS vectors available for Rosaceae fruit trees were reported so far. We present an effective VIGS system in apple, pear, and Japanese pear using Apple latent spherical virus (ALSV) vectors.

Results

Inoculation of ALSV vectors carrying a partial sequence of endogenous genes from apple [ribulose-1, 5-bisphosphate carboxylase small subunit (rbcS), alpha subunit of chloroplast chaperonin (CPN60a), elongation factor 1 alpha (EF-1a), or actin] to the cotyledons of seeds by a particle bombardment induced highly uniform knock-down phenotypes of each gene on the true leaves of seedlings from 2~3 weeks after inoculation. These silencing phenotypes continued for several months. Northern blot and RT-PCR analyses of leaves infected with ALSV containing a fragment of rbcS gene showed that the levels of rbcS-mRNA drastically decreased in the infected apple and pear leaves, and, in reverse, rbcS- siRNAs were generated in the infected leaves. In addition, some of apple seedlings inoculated with ALSV vector carrying a partial sequence of a TERMINAL FLOWER 1 gene of apple (MdTFL1) showed precocious flowering which is expected as a knock-down phenotype of the silencing of MdTFL1 gene.

Conclusions

The ALSV-based VIGS system developed have provides a valuable new addition to the tool box for functional genomics in apple, pear, and Japanese pear.
Mutation spectrum of MLL2 in a cohort of kabuki syndrome patients
Authors: Micale Lucia , Augello Bartolomeo , Fusco Carmela , Selicorni Angelo , Loviglio Maria N , Silengo Margherita Cirillo , Reymond Alexandre , Gumiero Barbara , Zucchetti Federica , D'Addetta Ester V , Belligni Elga , Calcagnì Alessia , Digilio Maria C , Dallapiccola Bruno , Faravelli Francesca , Forzano Francesca , Accadia Maria , Bonfante Aldo , Clementi Maurizio , Daolio Cecilia , Douzgou Sofia , Ferrari Paola , Fischetto Rita , Garavelli Livia , Lapi Elisabetta , Mattina Teresa , Melis Daniela , Patricelli Maria G , Priolo Manuela , Prontera Paolo , Renieri Alessandra , Mencarelli Maria A , Scarano Gioacchino , Monica Matteo della , Toschi Benedetta , Turolla Licia , Vancini Alessandra , Zatterale Adriana , Gabrielli Orazio , Zelante Leopoldo , Merla Giuseppe
Published: June 2011

Background

Kabuki syndrome (Niikawa-Kuroki syndrome) is a rare, multiple congenital anomalies/mental retardation syndrome characterized by a peculiar face, short stature, skeletal, visceral and dermatoglyphic abnormalities, cardiac anomalies, and immunological defects. Recently mutations in the histone methyl transferase MLL2 gene have been identified as its underlying cause.

Methods

Genomic DNAs were extracted from 62 index patients clinically diagnosed as affected by Kabuki syndrome. Sanger sequencing was performed to analyze the whole coding region of the MLL2 gene including intron-exon junctions. The putative causal and possible functional effect of each nucleotide variant identified was estimated by in silico prediction tools.

Results

We identified 45 patients with MLL2 nucleotide variants. 38 out of the 42 variants were never described before. Consistently with previous reports, the majority are nonsense or frameshift mutations predicted to generate a truncated polypeptide. We also identified 3 indel, 7 missense and 3 splice site.

Conclusions

This study emphasizes the relevance of mutational screening of the MLL2 gene among patients diagnosed with Kabuki syndrome. The identification of a large spectrum of MLL2 mutations possibly offers the opportunity to improve the actual knowledge on the clinical basis of this multiple congenital anomalies/mental retardation syndrome, design functional studies to understand the molecular mechanisms underlying this disease, establish genotype-phenotype correlations and improve clinical management.
A case of bowel entrapment after penetrating injury of the pelvis: don't forget the omentumplasty
Authors: Ritchie Ewan D , Veen Eelco J , Olsman Jan , Bosscha Koop
Published: June 2011

Bowel entrapment within a pelvic injury is rare and difficult to diagnose. Usually, it is diagnosed late because of concomitant abdominal injuries. It may present itself as an acute intestinal obstruction or, more commonly, as a prolonged or intermittent ileus. Therefore, one should be aware of this late complication and primarily take measures for avoiding bowel entrapment. This report describes an unusual case of bowel entrapment within a pelvic fracture after a penetrating injury, and discusses options for preventing such a complication.
The opinion of clinical staff regarding painfulness of procedures in pediatric hematology-oncology: an Italian survey
Authors: Po' Chiara , Benini Franca , Sainati Laura , Frigo Anna C , Cesaro Simone , Farina Maria I , Agosto Caterina
Published: June 2011

Background

Beliefs of caregivers about patient's pain have been shown to influence assessment and treatment of children's pain, now considered an essential part of cancer treatment. Painful procedures in hematology-oncology are frequently referred by children as the most painful experiences during illness. Aim of this study was to evaluate professionals' beliefs about painfulness of invasive procedures repeatedly performed in Pediatric Hemato-Oncology Units.

Methods

Physicians, nurses, psychologists and directors working in Hemato-Oncology Units of the Italian Association of Pediatric Hematology-Oncology (AIEOP) were involved in a wide-nation survey. The survey was based on an anonymous questionnaire investigating beliefs of operators about painfulness of invasive procedures (lumbar puncture, bone marrow aspirate and bone marrow biopsy) and level of pain management.

Results

Twenty-four directors, 120 physicians, 248 nurses and 22 psychologists responded to the questionnaire. The score assigned to the procedural pain on a 0-10 scale was higher than 5 in 77% of the operators for lumbar puncture, 97.5% for bone marrow aspiration, and 99.5% for bone marrow biopsy. The scores assigned by nurses differed statistically from those of the physicians and directors for the pain caused by lumbar puncture and bone marrow aspiration. Measures adopted for procedural pain control were generally considered good.

Conclusions

Invasive diagnostic-therapeutic procedures performed in Italian Pediatric Hemato-Oncology Units are considered painful by all the caregivers involved. Pain management is generally considered good. Aprioristically opinions about pain depend on invasiveness of the procedure and on the professional role.
The usefulness of magnetic resonance imaging of the hand and wrist in very early rheumatoid arthritis
Authors: Kosta Paraskevi E , Voulgari Paraskevi V , Zikou Anastasia K , Drosos Alexandros A , Argyropoulou Maria I
Published: June 2011

Introduction

Magnetic resonance imaging (MRI) was used to study the hand and wrist in very early rheumatoid arthritis (RA), and the results were compared with early and established disease.

Methods

Fifty-seven patients fulfilling the new American College of Rheumatology criteria for RA, 26 with very early RA (VERA), 18 with early RA (ERA), and 13 with established RA (ESTRA), (disease duration < 3 months, < 12 months, and > 12 months, respectively) were enrolled in the study. MRI of the dominant hand and wrist was performed by using fat-suppressed T2-weighted and plain and contrast-enhanced T1-weighted sequences. Evaluation of bone marrow edema, synovitis, and bone erosions was performed with the OMERACT RA MRI scoring system.

Results

Edema, erosions, and synovitis were present in VERA, and the prevalence was 100%, 96.15%, and 92.3%, respectively. Significant differences in edema and erosions were found between VERA and ESTRA (P < 0.05). No significant difference was found in synovitis.

Conclusions

Edema, erosions, and synovitis are findings of very early RA. MRI, by detecting these lesions, may play an important role in the management of these patients.
The SLC2A9 nonsynonymous Arg265His variant and gout: evidence for a population-specific effect on severity
Authors: Hollis-Moffatt Jade E , Gow Peter J , Harrison Andrew A , Highton John , Jones Peter BB , Stamp Lisa K , Dalbeth Nicola , Merriman Tony R
Published: June 2011

Introduction

The C allele of the nonsynonymous Arg265His (rs3733591) variant of SLC2A9 confers risk for gout in Han Chinese, Solomon Island and Japanese samples, with a stronger role in tophaceous gout. There is no evidence for an association with gout in Caucasian populations. In the present study, we tested rs3733591 for association with gout in New Zealand (NZ) Māori, Pacific Island and Caucasian samples.

Methods

Rs3733591 was genotyped across gout patients (n = 229, 232 and 327 NZ Māori, Pacific Island and Caucasian samples, respectively) and non-gout controls (n = 343, 174 and 638 Māori, Pacific Island and Caucasian samples, respectively). Further Caucasian sample sets consisting of 67 cases and 4,712 controls as well as 153 cases and 6,969 controls were obtained from the Framingham Heart Study and the Atherosclerosis Risk in Communities study, respectively. The Polynesian samples were analyzed according to Eastern and Western Polynesian ancestry.

Results

No evidence for risk conferred by the C allele of rs3733591 with gout was found in the sample sets of NZ Māori (odd ratio (OR) = 0.98, P = 0.86), Eastern Polynesians (OR = 0.99, P = 0.92), Western Polynesians (OR = 1.16, P = 0.36) or combined Caucasians (OR = 1.15, P = 0.13). The C allele was significantly overrepresented in Māori tophaceous cases compared to cases without tophi (OR = 2.21, P = 0.008), but not in the other ancestral groupings.

Conclusions

Noting that our study's power was limited for detecting weak genetic effects, we were unable to replicate associations of rs3733591 with gout in Eastern Polynesian, Western Polynesian and Caucasian samples. However, consistent with a previous study of Han Chinese and Solomon Island populations, our data suggest that rs3733591 could be a marker of severe gout in some populations. Our results also suggest that the effect of this variant is population-specific, further confirming population heterogeneity regarding the association of SLC2A9 with gout.
Health-related quality of life and physical recovery after a critical illness: a multi-centre randomised controlled trial of a home-based physical rehabilitation program
Authors: Elliott Doug , McKinley Sharon , Alison Jennifer , Aitken Leanne M , King Madeleine , Leslie Gavin D , Kenny Patricia , Taylor Penny , Foley Rachel , Burmeister Elizabeth
Published: June 2011

Introduction

Significant physical sequelae exist for some survivors of a critical illness. There are, however, few studies that have examined specific interventions to improve their recovery, and none have tested a home-based physical rehabilitation program incorporating trainer visits to participants' homes. This study was designed to test the effect of an individualised eight-week home-based physical rehabilitation program on recovery.

Methods

A multi-centre randomised controlled trial design was used. Adult intensive care patients (length of stay of at least 48 hours and mechanically ventilated for 24 hours or more) were recruited from 12 Australian hospitals between 2005 and 2008. Graded, individualised endurance and strength training intervention was prescribed over eight weeks, with three physical trainer home visits, four follow-up phone calls, and supported by a printed exercise manual. The main outcome measures were blinded assessments of physical function; SF-36 physical function (PF) scale and six-minute walk test (6MWT), and health-related quality of life (SF-36) conducted at 1, 8 and 26 weeks after hospital discharge.

Results

Of the 195 participants randomised, 183, 173 and 161 completed the 1, 8 and 26 weeks assessments, respectively. Study groups were similar at Week 1 post-hospital; for the intervention and control groups respectively, mean norm-based PF scores were 27 and 29 and the 6MWT distance was 291 and 324 metres. Both groups experienced significant and clinically important improvements in PF scores and 6MWT distance at 8 weeks, which persisted at 26 weeks. Mixed model analysis showed no significant group effects (P = 0.84) or group by time interactions (P = 0.68) for PF. Similar results were found for 6MWT and the SF-36 summary scores.

Conclusions

This individualised eight-week home-based physical rehabilitation program did not increase the underlying rate of recovery in this sample, with both groups of critically ill survivors improving their physical function over the 26 weeks of follow-up. Further research should explore improving effectiveness of the intervention by increasing exercise intensity and frequency, and identifying individuals who would benefit most from this intervention.

Trial registration

Australia and New Zealand Clinical Trials Register ACTRN12605000166673
Critical care services and the H1N1 (2009) influenza epidemic in Australia and New Zealand in 2010: the impact of the second winter epidemic
Authors:   The ANZIC Influenza Investigators
Published: June 2011

Introduction

During the first winter of exposure, the H1N1 2009 influenza virus placed considerable strain on intensive care unit (ICU) services in Australia and New Zealand (ANZ). We assessed the impact of the H1N1 2009 influenza virus on ICU services during the second (2010) winter, following the implementation of vaccination.

Methods

A prospective, cohort study was conducted in all ANZ ICUs during the southern hemisphere winter of 2010. Data on demographic and clinical characteristics, including vaccination status and outcomes, were collected. The characteristics of patients admitted during the 2010 and 2009 seasons were compared.

Results

From 1 June to 15 October 2010, there were 315 patients with confirmed influenza A, of whom 283 patients (90%) had H1N1 2009 (10.6 cases per million inhabitants; 95% confidence interval (CI), 9.4 to 11.9) which was an observed incidence of 33% of that in 2009 (P < 0.001). The maximum daily ICU occupancy was 2.4 beds (95% CI, 1.8 to 3) per million inhabitants in 2010 compared with 7.5 (95% CI, 6.5 to 8.6) in 2009, (P < 0.001). The onset of the epidemic in 2010 was delayed by five weeks compared with 2009. The clinical characteristics were similar in 2010 and 2009 with no difference in the age distribution, proportion of patients treated with mechanical ventilation, duration of ICU admission, or hospital mortality. Unlike 2009 the incidence of critical illness was significantly greater in New Zealand (18.8 cases per million inhabitants compared with 9 in Australia, P < 0.001). Of 170 patients with known vaccination status, 26 (15.3%) had been vaccinated against H1N1 2009.

Conclusions

During the 2010 ANZ winter, the impact of H1N1 2009 on ICU services was still appreciable in Australia and substantial in New Zealand. Vaccination failure occurred.
Acute kidney injury is an independent risk factor for pediatric intensive care unit mortality, longer length of stay and prolonged mechanical ventilation in critically ill children: a two-center retrospective cohort study
Authors: Alkandari Omar , Eddington K Allen , Hyder Ayaz , Gauvin France , Ducruet Thierry , Gottesman Ronald , Phan Véronique , Zappitelli Michael
Published: June 2011

Introduction

In adults, small (< 50%) serum creatinine (SCr) increases predict mortality. It is unclear whether different baseline serum creatinine (bSCr) estimation methods affect findings of acute kidney injury (AKI)-outcome associations. We characterized pediatric AKI, evaluated the effect of bSCr estimation approaches on AKI-outcome associations and evaluated the use of small SCr increases to predict AKI development.

Methods

We conducted a retrospective cohort database study of children (excluding postoperative cardiac or renal transplant patients) admitted to two pediatric intensive care units (PICUs) for at least one night in Montreal, QC, Canada. The AKI definition was based on the Acute Kidney Injury Network staging system, excluding the requirement of SCr increase within 48 hours, which was impossible to evaluate on the basis of our data set. We estimated bSCr two ways: (1) the lowest SCr level in the three months before admission or the average age- and gender-based norms (the standard method) or (2) by using average norms in all patients. Outcomes were PICU mortality and length of stay as well as required mechanical ventilation. We used multiple logistic regression analysis to evaluate AKI risk factors and the association between AKI and mortality. We used multiple linear regression analysis to evaluate the effect of AKI on other outcomes. We calculated diagnostic characteristics for early SCr increase (< 50%) to predict AKI development.

Results

Of 2,106 admissions (mean age ± SD = 5.0 ± 5.5 years; 47% female), 377 patients (17.9%) developed AKI (using the standard bSCr method) during PICU admission. Higher Pediatric Risk of Mortality score, required mechanical ventilation, documented infection and having a bSCr measurement were independent predictors of AKI development. AKI was associated with increased mortality (adjusted odds ratio (OR) = 3.7, 95% confidence interval (95% CI) = 2.1 to 6.4, using the standard bSCr method; OR = 4.5, 95% CI = 2.6 to 7.9, using normative bSCr values in all patients). AKI was independently associated with longer PICU stay and required mechanical ventilation. In children with no admission AKI, the initial percentage SCr increase predicted AKI development (area under the curve = 0.67, 95% CI = 0.60 to 0.74).

Conclusions

AKI is associated with increased mortality and morbidity in critically ill children, regardless of the bSCr used. Paying attention to small early SCr increases may contribute to early AKI diagnosis in conjunction with other new AKI biomarkers.