bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterizing rapid, activity‐linked conformational transitions in proteins via sub‐second hydrogen deuterium exchange mass spectrometry
Authors: Resetca Diana , Wilson Derek J.
Published: May 2013

This review outlines the application of time‐resolved electrospray ionization mass spectrometry (TRESI‐MS) and hydrogen‐deuterium exchange (HDX) to study rapid, activity‐linked conformational transitions in proteins. The method is implemented on a microfluidic chip which incorporates all sample‐handling steps required for a ‘bottom‐up’ HDX workflow: a capillary mixer for sub‐second HDX labeling, a static mixer for HDX quenching, a microreactor for rapid protein digestion, and on‐chip electrospray. By combining short HDX labeling pulses with rapid digestion, this approach provides a detailed characterization of the structural transitions that occur during protein folding, ligand binding, post‐translational modification and catalytic turnover in enzymes. This broad spectrum of applications in areas largely inaccessible to conventional techniques means that microfluidics‐enabled TRESI‐MS/HDX is a unique and powerful approach for investigating the dynamic basis of protein function.
Multimeric and differential binding of CIN85/CD2AP with two atypical proline‐rich sequences from CD2 and Cbl‐b*
Authors: Ceregido M. Angeles , Garcia‐Pino Abel , Ortega‐Roldan Jose L. , Casares Salvador , López Mayorga Obdulio , Bravo Jeronimo , van Nuland Nico A. J. , Azuaga Ana I.
Published: May 2013

The CD2AP (CD2‐associated protein) and CIN85 (Cbl‐interacting protein of 85 kDa) adaptor proteins each employ three Src homology 3 (SH3) domains to cluster protein partners and ensure efficient signal transduction and down‐regulation of tyrosine kinase receptors. Using NMR, isothermal titration calorimetry and small‐angle X‐ray scattering methods, we have characterized several binding modes of the N‐terminal SH3 domain (SH3A) of CD2AP and CIN85 with two natural atypical proline‐rich regions in CD2 (cluster of differentiation 2) and Cbl‐b (Casitas B‐lineage lymphoma), and compared these data with previous studies and published crystal structures. Our experiments show that the CD2AP‐SH3A domain forms a type II dimer with CD2 and both type I and type II dimeric complexes with Cbl‐b. Like CD2AP, the CIN85‐SH3A domain forms a type II complex with CD2, but a trimeric complex with Cbl‐b, whereby the type I and II interactions take place at the same time. Together, these results explain how multiple interactions among similar SH3 domains and ligands produce a high degree of diversity in tyrosine kinase, cell adhesion or T‐cell signaling pathways.

Structured digital abstract

CIN85 and Cbl-b bind by nuclear magnetic resonance (View Interaction: 1, 2)

CD2 and CIN85 bind by nuclear magnetic resonance (View interaction)

CD2 and CD2AP bind by nuclear magnetic resonance (View interaction)

Cbl-b and CD2AP bind by nuclear magnetic resonance (View Interaction: 1, 2)

Cbl-b and CIN85 bind by isothermal titration calorimetry (View Interaction: 1, 2, 3, 4)

CIN85 and Cbl-b bind by X-ray scattering (View Interaction: 1, 2)

CIN85 and CD2 bind by isothermal titration calorimetry (View interaction)

CD2 and CD2AP bind by X-ray scattering (View interaction)

CD2AP and CD2 bind by isothermal titration calorimetry (View interaction)

CD2 and CIN85 bind by X-ray scattering (View interaction)

CD2AP and Cbl-b bind by isothermal titration calorimetry (View Interaction: 1, 2, 3, 4, 5)
The hallmarks of Parkinson's disease
Authors: Antony Paul M. A. , Diederich Nico J. , Krüger Rejko , Balling Rudi
Published: May 2013

Since the discovery of dopamine as a neurotransmitter in the 1950s, Parkinson's disease (PD) research has generated a rich and complex body of knowledge, revealing PD to be an age‐related multifactorial disease, influenced by both genetic and environmental factors. The tremendous complexity of the disease is increased by a nonlinear progression of the pathogenesis between molecular, cellular and organic systems. In this minireview, we explore the complexity of PD and propose a systems‐based approach, organizing the available information around cellular disease hallmarks. We encourage our peers to adopt this cell‐based view with the aim of improving communication in interdisciplinary research endeavors targeting the molecular events, modulatory cell‐to‐cell signaling pathways and emerging clinical phenotypes related to PD.
Are silicate‐rich inclusions in spodumene crystallized aliquots of boundary layer melt?
Authors: Anderson A. J.
Published: May 2013

The constitutional zone refining model for the internal crystallization of granitic pegmatites stipulates that a highly fluxed and incompatible element‐enriched boundary layer melt evolves at the surface of crystal growth fronts in undercooled granitic melts. It has been argued that silicate‐rich inclusions in primary spodumene and petalite from the Tanco pegmatite are the crystallized products of an entrapped boundary layer melt. However, inclusions of identical character and composition to those in primary spodumene are also abundant in the spodumene component of SQI (spodumene + quartz intergrowths) that formed by the isochemical breakdown of petalite. Both textural types of spodumene at Tanco host fluid inclusion assemblages that display highly variable proportions of solid phases (dominantly cookeite, quartz, and zabuyelite) and a low salinity (approximately 6 wt% NaCl equivalent) aqueous fluid. The average major element content of three fluid inclusion assemblages is approximately Li2O (3 wt%), Al2O3 (11 wt%), SiO2 (30 wt%), CO2 (3 wt%) and H2O (53 wt%). The entrapment of these fluid inclusions is temporally and spatially unrelated to the flux‐rich melt that may have developed at the primary crystallization front of the pegmatite. Petrographic evidence and the mineralogical and chemical composition of spodumene‐hosted inclusions suggest that inclusions in spodumene are the products of hydrothermal dissolution of the host and the precipitation of quartz, cookeite, and zabuyelite prior to and during necking.
“Metakaolin‐Slag‐Clinker Blends.” The Role of Na+ or K+ as Alkaline Activators of Theses Ternary Blends
Authors: Fernández‐Jiménez Ana , Zibouche Fatima , Boudissa Nassima , García‐Lodeiro Ines , Abadlia Mohamed Tahar , Palomo Angel
Published: May 2013

This article reports on research into the use of solid alkalis (Na2CO3 and K2CO3) as activators to obtain hybrid cement (cement whose hydration generates a mix of C–A–S–H and (N,C)–A–S–H gels) from a blend of 20% clínker + 40% blast furnace slag + 40% metakaolin. More specifically, the study aimed to determine the effect of activator dosage (5 and 8 wt%) and type of alkaline cation (Na+ or K+) on the 2‐ and 28‐d mechanical strength of the end materials. The findings showed that the highest mechanical strength values were obtained with 5% Na2CO3. According to the XRD, NMR, and SEM/EDX analyses conducted on the reaction products, the alkalinity and solubilized chemical species generated by adding 5% Na2CO3 to the system yielded a mix of (N,C)–A–S–H and C–A–S–H cementitious gels as the main reaction products. The secondary reaction products included metastable (3CaO·Al2O3·CaCO3·11H2O‐type) carboaluminates that evolved into the calcite or vaterite forms of calcium carbonate. When K2CO3 was used (instead of Na2CO3), a (3CaO·Al2O3·0.5Ca(OH)2·0.5CaCO3·11H2O‐type) hemicarboaluminate also formed. The study also revealed that Na+ favors coagulation/precipitation more effectively than K+, generating gels with a wider range of Qn species.
Limits to the Stability of the Amorphous Nature of Polymer‐Derived HfSiCNO Compounds
Authors: Terauds Kalvis , Raj Rishi
Published: May 2013

Substituting silicon by transition metals in polymer‐derived ceramics (PDCs) holds the potential for a new class of polymer‐derived ceramics for ultrahigh‐temperature structural applications. We present experiments that show that the solid solubility of HfO2 extends to Hf/Si ratio of <0.22. The materials are synthesized from (miscible) organic precursors. Similar to silicon‐based materials they remain amorphous after pyrolysis at 1000°C. Small‐angle X‐ray scattering and Raman spectra remain essentially unaltered. It is postulated that Hf, like Si, forms mixed‐bond tetrahedra with C, O, and N. The difference in the enthalpy of Hf‐based, and Si‐centered tetrahedra is calculated using single‐bond energies, reinforcing the feasibility of substituting Si with Hf or with Zr atoms. Such polymer‐based HfSiCNO compounds made directly from liquid organics, by a simple manufacturing process, may also be relevant to nanoscale dielectrics with low leakage electric charge in microelectronics applications.
Rheological Control of the Coffee Stain Effect for Inkjet Printing of Ceramics
Authors: Friederich A. , Binder J. R. , Bauer W.
Published: May 2013

The rheology of inkjet printing inks must be well controlled in order to be able to form small droplets. One solution is to use low volume fraction dispersed suspensions, but this leads to a common problem during drying called the coffee stain effect. It is caused by particle migration from the center to the edge of a drying drop and leads to nonuniform printed structures. This article describes an approach, to suppress the coffee stain effect by a sufficiently fast increase in viscosity after deposition. Due to the viscosity limitations during printing, inks with tailored rheology and drying behavior need to be developed. Ceramic inks were prepared and printed. First, a binder was added to study the influence of viscosity on printability and the coffee stain effect. Second, the use of a high vapor pressure solvent for faster drying was investigated. Eventually, an ink with the combination of binder and fast drying agent was prepared. This ink showed a considerable decrease in drying time as well as a rapid increase in viscosity after deposition and was suitable to completely suppress the coffee stain effect. Plateau‐like structures were achieved by adapting the drying temperature to permit particle movement to a certain degree.
Directional protein secretion by the retinal pigment epithelium: roles in retinal health and the development of age‐related macular degeneration
Authors: Kay Paul , Yang Yit C. , Paraoan Luminita
Published: May 2013

The structural and functional integrity of the retinal pigment epithelium (RPE) is fundamental for maintaining the function of the neuroretina. These specialized cells form a polarized monolayer that acts as the retinal–blood barrier, separating two distinct environments with highly specialized functions: photoreceptors of the neuroretina at the apical side and Bruch's membrane/highly vascularized choriocapillaris at the basal side. The polarized nature of the RPE is essential for the health of these two regions, not only in nutrient and waste transport but also in the synthesis and directional secretion of proteins required in maintaining retinal homoeostasis and function. Although multiple malfunctions within the RPE cells have been associated with development of age‐related macular degeneration (AMD), the leading cause of legal blindness, clear causative processes have not yet been conclusively characterized at the molecular and cellular level. This article focuses on the involvement of directionally secreted RPE proteins in normal functioning of the retina and on the potential association of incorrect RPE protein secretion with development of AMD. Understanding the importance of RPE polarity and the correct secretion of essential structural and regulatory components emerge as critical factors for the development of novel therapeutic strategies targeting AMD.
In vitro chipping behaviour of all‐ceramic crowns with a zirconia framework and feldspathic veneering: comparison of CAD/CAM‐produced veneer with manually layered veneer
Authors: Schmitter M. , Mueller D. , Rues S.
Published: May 2013

The purpose of this in vitro study was to assess the breaking load of zirconia‐based crowns veneered with either CAD/CAM‐produced or manually layered feldspathic ceramic. Thirty‐two identical zirconia frameworks (Sirona inCoris ZI, mono L F1), 0·6 mm thick with an anatomically shaped occlusal area, were constructed (Sirona inLab 3.80). Sixteen of the crowns were then veneered by the use of CAD/CAM‐fabricated feldspathic ceramic (CEREC Bloc, Sirona) and 16 by the use of hand‐layered ceramic. The CAD/CAM‐manufactured veneer was attached to the frameworks by the use of Panavia 2.0 (Kuraray). Half of the specimens were loaded until failure without artificial ageing; the other half of the specimens underwent thermal cycling and cyclic loading (1·2 million chewing cycles, force magnitude Fmax = 108 N) before the assessment of the ultimate load. To investigate the new technique further, finite element (FE) computations were conducted on the basis of the original geometry. Statistical assessment was made by the use of non‐parametric tests. Initial breaking load was significantly higher in the hand‐layered group than in the CAD/CAM group (mean: 1165·86 N versus 395·45 N). During chewing simulation, however, 87·5% (7/8) of the crowns in the hand‐layered group failed, whereas no crown in the CAD/CAM group failed. The CAD/CAM‐produced veneer was significantly less sensitive to ageing than the hand‐layered veneer.
Discrepancies About Adolescent Relationships as a Function of Informant Attachment and Depressive Symptoms
Authors: Ehrlich Katherine B. , Cassidy Jude , Lejuez Carl W. , Daughters Stacey B.
Published: May 2013

Developmental scientists studying interpersonal relationships often find that informants disagree in their reports, and two theoretical perspectives suggest that these discrepancies may be predictable. In two studies of adolescents' interpersonal relationships, we examined two factors that may predict the absolute magnitude and the direction of the discrepancies in reports about several types of relationships. Specifically, we examined informants' depressive symptoms and attachment as predictors of absolute and directional discrepancies in reports about (a) adolescents' relationships with peers, (b) parent–adolescent relationships, and (c) adolescents' friendships. Findings revealed that informant depressive symptoms sometimes were associated with discrepancies. In contrast, informant attachment more consistently accounted for absolute and directional discrepancies.
Assessment of the quality of reporting in abstracts of systematic reviews with meta‐analyses in periodontology and implant dentistry
Authors: Faggion C. M. , Liu J. , Huda F. , Atieh M.
Published: May 2013

Background

Proper scientific reporting is necessary to ensure the correct interpretation of study results by readers. The main objective of this study was to assess the quality of reporting in abstracts of systematic reviews (SRs) with meta‐analyses in periodontology and implant dentistry. Differences in reporting of abstracts in Cochrane and paper‐based reviews were also assessed.

Methods

The PubMed electronic database and the Cochrane database for SRs were searched on November 11, 2012, independently and in duplicate, for SRs with meta‐analyses related to interventions in periodontology and implant dentistry. Assessment of the quality of reporting was performed independently and in duplicate, taking into account items related to the effect direction, numerical estimates of effect size, measures of precision, probability and consistency.

Results

We initially screened 433 papers and included 146 (127 paper‐based and 19 Cochrane reviews, respectively). The direction of evidence was reported in two‐thirds of the abstracts while strength of evidence and measure of precision (i.e., confidence interval) were reported in less than half the selected abstracts. Measures of consistency such as I2 statistics were reported in only 5% of the selected sample of abstracts. Cochrane abstracts reported the limitations of evidence and precision better than paper‐based ones. Two items (“meta‐analysis” in title and abstract, respectively), were nevertheless better reported in paper‐based abstracts.

Conclusion

Abstracts of SRs with meta‐analyses in periodontology and implant dentistry currently have no uniform standard of reporting, which may hinder readers’ understanding of study outcomes.
The relationship between root concavities in first premolars and chronic periodontitis
Authors: Zhao H. , Wang H. , Pan Y. , Pan C. , Jin X.
Published: May 2013

Objective

To evaluate the significance of first premolar root concavity on clinical indices of chronic periodontal disease and alveolar bone defects.

Methods

Three‐dimensional reconstruction by cone beam computed tomography was used to observe root surface anatomy and the type of alveolar bone defect seen in the mesial and distal sites of 272 first premolars from 99 patients who had presented with chronic periodontitis. Periodontal clinical indicators at each site were measured using a Florida Probe Corporation (Gainesville, FL, USA).

Results

The incidence of mesial and distal root concavities of the maxillary first premolars was 100% and 39.3% respectively, and in the mandibular, the incidence was 42.5% and 31.3% respectively. The distributions of the different types of concavities in terms of both age and gender of the patients were not statistically significant. The mean probing depth and clinical attachment loss of the first premolars with root concavities were significantly higher than those without concavity (p < 0.05). Plaque accumulation was significantly different in the premolars with/without root concavities (p < 0.001). The type of alveolar bone defects with concavities was significantly different from those without concavities (p < 0.05). Ramp shape bone defects were dominant for teeth without concavities, while crater shape was seen for teeth with concavities.

Conclusion

Root concavities of the first premolars were associated with periodontal disease, and the type of interproximal alveolar bone defect. Root concavities may be important in contributing to local periodontal disease of the first premolars.
Clinical usefulness of a guideline‐based screening tool for the diagnosis of allergic rhinitis in asthmatics: The Self Assessment of Allergic Rhinitis and Asthma questionnaire
Authors: Hojo Masayuki , Ohta Ken , Iikura Motoyasu , Mizutani Tomonori , Hirashima Junko , Sugiyama Haruhito
Published: May 2013

Background and objective

Allergic rhinitis (AR) is common in asthmatic patients and may impair asthma control. However, this comorbidity is frequently missed. A simple test is needed to enable physicians to diagnose and evaluate the severity of both diseases. The Self Assessment of Allergic Rhinitis and Asthma (SACRA) questionnaire, which GINA and ARIA Japan committees developed in 2011, consist of questionnaires based on GINA and ARIA guidelines for the diagnosis and severity of AR and asthma, and a visual analogue scale (VAS) to evaluate the severity of both diseases. Our objective was to investigate the clinical usefulness of SACRA as a patient‐based screening tool for identifying asthmatic patients with AR.

Methods

SACRA, ACT (Asthma Control Test) and serum IgE RAST were performed in asthmatic patients. The correlation between SACRA and other parameters were analyzed.

Results

Four hundred twenty asthmatic patients were enrolled. Among 168 subjects who self‐reported no concomitant AR, 76 asthmatics scored one or more symptoms on SACRA. Eventually, 32 of these 76 subjects were diagnosed with AR by physicians based on laboratory data or physical examinations by ear, nose and throat specialists. The sensitivity and specificity of SACRA for the diagnosis of AR were 92% and 66%, respectively. The estimated prevalence of AR among asthmatics was 66%, almost identical to that of the previous nationwide study in Japan. The level of asthma control assessed by the VAS on SACRA and the ACT score showed a strong correlation (r = −0.700, P < 0.001).

Conclusions

SACRA may be a clinically useful tool for identifying bronchial asthma patients with AR.
Attachment Patterns in Medically Serious Suicide Attempts: The Mediating Role of Self‐Disclosure and Loneliness
Authors: Levi‐Belz Yossi , Gvion Yari , Horesh Netta , Apter Alan
Published: May 2013

Although the study of medically serious suicide behavior is an important strategy for understanding the nature of suicide, little is known about its underlying psychological mechanisms. This gap is addressed here by applying insights from attachment theory to severe suicidal behavior. The results show that both anxious and avoidant attachment patterns predict medical lethality. Path analysis indicated that interpersonal difficulties mediated the paths between insecure attachment patterns and lethality of suicide attempts. These results suggest that the psychological mechanisms of medically serious suicide behavior involve high levels of mental pain amplified by insecure attachment patterns and interpersonal difficulties. Implications for prevention and therapeutic intervention strategies are discussed.
PpeTAC1 promotes the horizontal growth of branches in peach trees and is a member of a functionally conserved gene family found in diverse plants species
Authors: Dardick Chris , Callahan Ann , Horn Renate , Ruiz Karina B. , Zhebentyayeva Tetyana , Hollender Courtney , Whitaker Michael , Abbott Albert , Scorza Ralph
Published: May 2013

Trees are capable of tremendous architectural plasticity, allowing them to maximize their light exposure under highly competitive environments. One key component of tree architecture is the branch angle, yet little is known about the molecular basis for the spatial patterning of branches in trees. Here, we report the identification of a candidate gene for the br mutation in Prunus persica (peach) associated with vertically oriented growth of branches, referred to as ‘pillar’ or ‘broomy’. Ppa010082, annotated as hypothetical protein in the peach genome sequence, was identified as a candidate gene for br using a next generation sequence‐based mapping approach. Sequence similarity searches identified rice TAC1 (tiller angle control 1) as a putative ortholog, and we thus named it PpeTAC1. In monocots, TAC1 is known to lead to less compact growth by increasing the tiller angle. In Arabidopsis, an attac1 mutant showed more vertical branch growth angles, suggesting that the gene functions universally to promote the horizontal growth of branches. TAC1 genes belong to a gene family (here named IGT for a shared conserved motif) found in all plant genomes, consisting of two clades: one containing TAC1‐like genes; the other containing LAZY1, which contains an EAR motif, and promotes vertical shoot growth in Oryza sativa (rice) and Arabidopsis through influencing polar auxin transport. The data suggest that IGT genes are ancient, and play conserved roles in determining shoot growth angles in plants. Understanding how IGT genes modulate branch angles will provide insights into how different architectural growth habits evolved in terrestrial plants.
A novel structural effector from rust fungi is capable of fibril formation
Authors: Kemen Eric , Kemen Ariane , Ehlers Andreas , Voegele Ralf , Mendgen Kurt
Published: May 2013

It has been reported that filament‐forming surface proteins such as hydrophobins are important virulence determinants in fungi and are secreted during pathogenesis. Such proteins have not yet been identified in obligate biotrophic pathogens such as rust fungi. Rust transferred protein 1 (RTP1p), a rust protein that is transferred into the host cytoplasm, accumulates around the haustorial complex. To investigate RTP1p structure and function, we used immunocytological, biochemical and computational approaches. We found that RTP1p accumulates in protuberances of the extra‐haustorial matrix, a compartment that surrounds the haustorium and is separated from the plant cytoplasm by a modified host plasma membrane. Our analyses show that RTP1p is capable of forming filamentous structures in vitro and in vivo. We present evidence that filament formation is due to β–aggregation similar to what has been observed for amyloid‐like proteins. Our findings reveal that RTP1p is a member of a new class of structural effectors. We hypothesize that RTP1p is transferred into the host to stabilize the host cell and protect the haustorium from degradation in later stages of the interaction. Thus, we provide evidence for transfer of an amyloid‐like protein into the host cell, which has potential for the development of new resistance mechanisms against rust fungi.
Adjuvant therapy with carboplatin and pamidronate for canine appendicular osteosarcoma
Authors: Kozicki A. R. , Robat C. , Chun R. , Kurzman I. D.
Published: May 2013

Amputation and chemotherapy are the mainstay of treatment for canine appendicular osteosarcoma (OSA).In vitrostudies have demonstrated anti‐tumour activity of pamidronate against canineOSA. The purpose of this study was to assess the safety of adding pamidronate to standard post‐operative carboplatin chemotherapy in 17 dogs with appendicularOSAtreated with limb amputation. Median disease‐free interval (DFI) and median survival time (MST) were evaluated as secondary endpoints. Incidence of side effects and treatment outcomes were compared to 14 contemporary control patients treated with carboplatin alone. There were no identified side effects to the pamidronate treatment. The medianDFIfor the study group was 185 days compared to 172 days for the control group (P= 0.90). TheMSTof the study group was 311 days compared to 294 days for the control group (P= 0.89). Addition of pamidronate to carboplatin chemotherapy for the treatment of canine appendicularOSAis safe and does not impair efficacy of standard carboplatin treatment.
The effect of increased centrifugation temperature on the quality of red‐blood‐cell concentrates of automated whole blood processing
Authors: Weinigel C. , Rummler S. , Barz D.
Published: May 2013

Background and Objectives

There are manual and automated methods to separate whole blood (WB) available. The Atreus whole blood processing system is an automated method, which combines centrifugation and expression of components into a single device. A major difference to conventional methods is that centrifugation temperature is not controlled at 22°C. The aim of this study was to examine the influence of increased centrifugation temperatures on the quality of red‐blood‐cell concentrates (RCC) after active cooling of WB prior to processing.

Materials and Methods

A total of 28 WB were processed: 16 at centrifugation temperatures of up to 28°C (1st protocol) and 12 at 34°C (2nd protocol). RCC quality parameters were tested weekly for 42 days.

Results

Red‐blood‐cell concentrates (RCC) quality complied with the European and German guidelines. Haemolysis was not significantly different throughout storage. Significant statistical differences were detected between both protocols in potassium concentration at the end of storage and in ATP levels at the day of processing.

Conclusion

Centrifugation temperatures of up to 34°C are well tolerated by the red blood cells with minimal interference with the RCC quality parameters.
Sensitivity of assays for the detection of HPA‐1a antibodies: results of an international workshop demonstrating the impact of cation chelation from integrin αIIbβ3 on three widely used assays
Authors: Allen D. L. , Metcalfe P. , Kaplan C. , Kekomaki R. , de Haas M. , Yusuf R. , Ouwehand W. H.
Published: May 2013

Background and Objectives

HPA‐1a antibodies account for 70–80% of cases of fetal–neonatal alloimmune thrombocytopenia (FNAIT) in Caucasians. However, numerous workshops have demonstrated variability in their detection. We recently showed that exposure of αIIbβ3 to ethylene diamine tetraacetic acid (EDTA) affected binding of many anti‐αIIbβ3 monoclonal, and HPA‐1a allo‐, antibodies; this adversely affected sensitivity of the monoclonal antibody‐specific immobilization of platelet antigens (MAIPA) assay and indirect platelet immunofluorescence test (PIFT). This study presents results from an international workshop studying the impact of cation chelation on HPA‐1a antibody detection in routine diagnostic laboratories.

Materials and Methods

Serum and EDTA‐anticoagulated plasma samples containing anti‐HPA‐1a were distributed to 39 laboratories. Participants were asked to detect and identify any HPA antibodies present.

Results

2/39 (5·1%) participants were able to detect and identify anti‐HPA‐1a in the serum, but not in the plasma sample. EDTA plasma reduced MAIPA assay sensitivity by ≥20% in 17/24 (70·8%) laboratories and by ≥50% in 9/24 (37·5%) when using HPA‐1a1a platelets (mean: 27·7%, range 0–85·1%); when using HPA‐1a1b platelets 3/4 (75%), participants reported ≥50% loss of sensitivity (mean 65·6%, range 0–96·6%). A small but significant increase in optical densities was observed in antigen capture ELISA assays when using plasma (mean difference: 0·081, P < 0·01). Insufficient PIFT data were returned to draw firm conclusions.

Conclusion

Use of EDTA plasma significantly affects the sensitivity of the MAIPA assay and can affect detection of even potent, FNAIT‐causing examples of anti‐HPA‐1a. These data highlight the importance of use of αIIbβ3 in an appropriate conformation for the sensitive detection of anti‐HPA‐1a.
Barriers and motivators to blood donation among university students in Japan: development of a measurement tool
Authors: Ngoma A. M. , Goto A. , Yamazaki S. , Machida M. , Kanno T. , Nollet K. E. , Ohto H. , Yasumura S.
Published: May 2013

Background and Objectives

Despite growing demand for transfusion, the number of voluntary young blood donors has steadily decreased over recent years in Japan. This study aimed to develop an easy‐to‐use survey tool to assess barriers and motivators to blood donation among Japanese university students.

Materials and Methods

We conducted cross‐sectional studies at two universities in Fukushima Prefecture, Japan, in December 2011 (Stage 1) and February 2012 (Stage 2) using self‐administered questionnaires. A short list of motivators and barriers to blood donation was developed from the open‐ended questions asked of 50 students in Stage 1. In the Stage 2, we asked 105 students how important these items were when they decided whether or not to donate blood. Items showing a significant difference between donors and non‐donors were kept in the final list.

Results

Overall, 56% of the 100 participants analysed in Stage 2 were men, and ages ranged from 19 to 24 with a median of 20 years. Comparison of motivators and barriers between donors and non‐donors revealed that only barrier item 8 (‘Frightened by blood donation’) showed a significant difference (P = 0·0006) in an expected direction and with a consistency between two universities.

Conclusions

This study identified fear as being the most significant barrier to blood donation among Japanese university students, which could be used as a single convenient indicator to assess their readiness to donate. More academic and clinical efforts are needed to understand and address students' fear towards blood donation in order to increase the donor pool in Japan.
Blood Pressure, Artery Size, and Artery Compliance Parallel Bone Size and Strength in Mice With Differing Ece1 Expression
Authors: Wang Zhijie , Kristianto Jasmin , Yen Ooi Chen , Johnson Michael G. , Litscher Suzanne J. , Pugh Thomas D. , Sandhu Gurpreet , Chesler Naomi C. , Blank Robert D.
Published: May 2013

The recombinant congenic mouse strains HcB-8 and HcB-23 differ in femoral shape, size, and strength, with HcB-8 femora being more gracile, more cylindrical, weaker, and having higher Young's modulus. In previous work, we mapped a robust, pleiotropic quantitative trait locus for these bone traits. Ece1, encoding endothelin converting enzyme 1, is a positional candidate gene for this locus, and was less expressed in HcB-8 bone. We hypothesized that the same genetic factors would impose analogous developmental trajectories on arteries to those in bones. Cardiovascular hemodynamics and biomechanics of carotids were measured in adult HcB-8 and HcB-23 mice. Biological differences in heart and arteries were examined at mRNA and protein levels. As in bone, Ece1 expression was higher in HcB-23 heart and arteries (p < 0.05), and its expression was correlated with that of the endothelin B type receptor target Nos3, encoding endothelial nitric oxide synthase. HcB-8 mice had higher ambulatory blood pressure (p < 0.005) than HcB-23 mice. Ex vivo, at identical pressures, HcB-8 carotid arteries had smaller diameters and lower compliance (p < 0.05), but the same elastic modulus compared to HcB-23 carotid arteries. HcB-8 hearts were heavier than HcB-23 hearts (p < 0.01). HcB-8 has both small, stiff bones and small, stiff arteries, lower expression of Ece1 and Nos3, associated in each case with less favorable function. These findings suggest that endothelin signaling could serve as a nexus for the convergence of skeletal and vascular modeling, providing a potential mechanism for the epidemiologic association between skeletal fragility and atherosclerosis.
A multimodality vascular imaging phantom of an abdominal aortic aneurysm with a visible thrombus
Authors: Allard Louise , Soulez Gilles , Chayer Boris , Qin Zhao , Roy David , Cloutier Guy
Published: May 2013

Purpose:

With the continuous development of new stent grafts and implantation techniques, it has now become technically feasible to treat abdominal aortic aneurysms (AAA) with challenging anatomy using endovascular repair with standard, fenestrated, or branched stent‐grafts.In vitro experimentations are very useful to improve stent‐graft design and conformability or imaging guidance for stent‐graft delivery or follow‐up. Vascular replicas also help to better understand the limitation of endovascular approaches in challenging anatomy and possibly improve surgical planning or training by practicing high risk clinical procedures in the laboratory to improve outcomes in the operating room. Most AAA phantoms available have a very basic anatomy, which is not representative of the clinical reality. This paper presents a method of fabrication of a realistic AAA phantom with a visible thrombus, as well as some mechanical properties characterizing such phantom.

Methods:

A realistic AAA geometry replica of a real patient anatomy taken from a multidetector computed tomography (CT) scan was manufactured. To demonstrate the multimodality imaging capability of this new phantom with a thrombus visible in magnetic resonance (MR) angiography, CT angiography (CTA), digital subtraction angiography (DSA), and ultrasound, image acquisitions with all these modalities were performed by using standard clinical protocols. Potential use of this phantom for stent deployment was also tested. A rheometer allowed defining hyperelastic and viscoelastic properties of phantom materials.

Results:

MR imaging measurements of SNR and CNR values on T1 and T2‐weighted sequences and MR angiography indicated reasonable agreement with published values of AAA thrombus and abdominal componentsin vivo. X‐ray absorption also lay within normal ranges of AAA patients and was representative of findings observed on CTA, fluoroscopy, and DSA. Ultrasound propagation speeds for developed materials were also in concordance with the literature for vascular and abdominal tissues.

Conclusions:

The mimicked abdominal tissues, AAA wall, and surrounding thrombus were developed to match imaging features ofin vivo MR, CT, and ultrasound examinations. This phantom should be of value for image calibration, segmentation, and testing of endovascular devices for AAA endovascular repair.
Nanoscale Effects of Caspofungin against Two Yeast Species, Saccharomyces cerevisiae and Candida albicans
Authors: Formosa C. , Schiavone M. , Martin-Yken H. , François J. M. , Duval R. E. , Dague E.
Published: May 2013

Saccharomyces cerevisiae
and
Candida albicans
are model yeasts for biotechnology and human health, respectively. We used atomic force microscopy (AFM) to explore the effects of caspofungin, an antifungal drug used in hospitals, on these two species. Our nanoscale investigation revealed similar, but also different, behaviors of the two yeasts in response to treatment with the drug. While administration of caspofungin induced deep cell wall remodeling in both yeast species, as evidenced by a dramatic increase in chitin and decrease in β-glucan content, changes in cell wall composition were more pronounced with
C. albicans
cells. Notably, the increase of chitin was proportional to the increase in the caspofungin dose. In addition, the Young modulus of the cell was three times lower for
C. albicans
cells than for
S. cerevisiae
cells and increased proportionally with the increase of chitin, suggesting differences in the molecular organization of the cell wall between the two yeast species. Also, at a low dose of caspofungin (i.e., 0.5× MIC), the cell surface of
C. albicans
exhibited a morphology that was reminiscent of cells expressing adhesion proteins. Interestingly, this morphology was lost at high doses of the drug (i.e., 4× MIC). However, the treatment of
S. cerevisiae
cells with high doses of caspofungin resulted in impairment of cytokinesis. Altogether, the use of AFM for investigating the effects of antifungal drugs is relevant in nanomedicine, as it should help in understanding their mechanisms of action on fungal cells, as well as unraveling unexpected effects on cell division and fungal adhesion.
Evaluation of Reduced Susceptibility to Quaternary Ammonium Compounds and Bisbiguanides in Clinical Isolates and Laboratory-Generated Mutants of Staphylococcus aureus
Authors: Furi Leonardo , Ciusa Maria Laura , Knight Daniel , Di Lorenzo Valeria , Tocci Nadia , Cirasola Daniela , Aragones Lluis , Coelho Joana Rosado , Freitas Ana Teresa , Marchi Emmanuela , Moce Laura , Visa Pilar , Northwood John Blackman , Viti Carlo , Borghi Elisa , Orefici Graziella ,   the BIOHYPO Consortium, Morrissey Ian , Oggioni Marco Rinaldo
Published: May 2013

The MICs and minimum bactericidal concentrations (MBCs) for the biocides benzalkonium chloride and chlorhexidine were determined against 1,602 clinical isolates of
Staphylococcus aureus
. Both compounds showed unimodal MIC and MBC distributions (2 and 4 or 8 mg/liter, respectively) with no apparent subpopulation with reduced susceptibility. To investigate further, all isolates were screened for
qac
genes, and 39 of these also had the promoter region of the NorA multidrug-resistant (MDR) efflux pump sequenced. The presence of
qacA
,
qacB
,
qacC
, and
qacG
genes increased the mode MIC, but not MBC, to benzalkonium chloride, while only
qacA
and
qacB
increased the chlorhexidine mode MIC. Isolates with a wild-type
norA
promoter or mutations in the
norA
promoter had similar biocide MIC distributions; notably, not all clinical isolates with
norA
mutations were resistant to fluoroquinolones.
In vitro
efflux mutants could be readily selected with ethidium bromide and acriflavine. Multiple passages were necessary to select mutants with biocides, but these mutants showed phenotypes comparable to those of mutants selected by dyes. All mutants showed changes in the promoter region of
norA
, but these were distinct from this region of the clinical isolates. Still, none of the
in vitro
mutants displayed fitness defects in a killing assay in
Galleria mellonella
larvae. In conclusion, our data provide an in-depth comparative overview on efflux in
S. aureus
mutants and clinical isolates, showing also that plasmid-encoded efflux pumps did not affect bactericidal activity of biocides. In addition, current
in vitro
tests appear not to be suitable for predicting levels of resistance that are clinically relevant.
Genetic Variability of Vancomycin-Resistant Enterococcus faecium and Enterococcus faecalis Isolates from Humans, Chickens, and Pigs in Malaysia
Authors: Getachew Yitbarek , Hassan Latiffah , Zakaria Zunita , Abdul Aziz Saleha
Published: May 2013

Vancomycin-resistant enterococci (VRE) have been reported to be present in humans, chickens, and pigs in Malaysia. In the present study, representative samples of VRE isolated from these populations were examined for similarities and differences by using the multilocus sequence typing (MLST) method. Housekeeping genes of
Enterococcus faecium
(
n
= 14) and
Enterococcus faecalis
(
n
= 11) isolates were sequenced and analyzed using the MLST databases eBURST and goeBURST. We found five sequence types (STs) of
E. faecium
and six STs of
E. faecalis
existing in Malaysia.
Enterococcus faecium
isolates belonging to ST203, ST17, ST55, ST79, and ST29 were identified, and
E. faecium
ST203 was the most common among humans. The MLST profiles of
E. faecium
from humans in this study were similar to the globally reported nosocomial-related strain lineage belonging to clonal complex 17 (CC17). Isolates from chickens and pigs have few similarities to those from humans, except for one isolate from a chicken, which was identified as ST203.
E. faecalis
isolates were more diverse and were identified as ST4, ST6, ST87, ST108, ST274, and ST244, which were grouped as specific to the three hosts.
E. faecalis
, belonging to the high-risk CC2 and CC87, were detected among isolates from humans. In conclusion, even though one isolate from a chicken was found clonal to that of humans, the MLST analysis of
E. faecium
and
E. faecalis
supports the findings of others who suggest VRE to be predominantly host specific and that clinically important strains are found mainly among humans. The infrequent detection of a human VRE clone in a chicken may in fact suggest a reverse transmission of VRE from humans to animals.
Arsenic Bioremediation by Biogenic Iron Oxides and Sulfides
Authors: Omoregie Enoma O. , Couture Raoul-Marie , Van Cappellen Philippe , Corkhill Claire L. , Charnock John M. , Polya David A. , Vaughan David , Vanbroekhoven Karolien , Lloyd Jonathan R.
Published: May 2013

Microcosms containing sediment from an aquifer in Cambodia with naturally elevated levels of arsenic in the associated groundwater were used to evaluate the effectiveness of microbially mediated production of iron minerals for
in situ
As remediation. The microcosms were first incubated without amendments for 28 days, and the release of As and other geogenic chemicals from the sediments into the aqueous phase was monitored. Nitrate or a mixture of sulfate and lactate was then added to stimulate biological Fe(II) oxidation or sulfate reduction, respectively. Without treatment, soluble As concentrations reached 3.9 ï¿½ 0.9 μM at the end of the 143-day experiment. However, in the nitrate- and sulfate-plus-lactate-amended microcosms, soluble As levels decreased to 0.01 and 0.41 ï¿½ 0.13 μM, respectively, by the end of the experiment. Analyses using a range of biogeochemical and mineralogical tools indicated that sorption onto freshly formed hydrous ferric oxide (HFO) and iron sulfide mineral phases are the likely mechanisms for As removal in the respective treatments. Incorporation of the experimental results into a one-dimensional transport-reaction model suggests that, under conditions representative of the Cambodian aquifer, the
in situ
precipitation of HFO would be effective in bringing groundwater into compliance with the World Health Organization (WHO) provisional guideline value for As (10 ppb or 0.13 μM), although soluble Mn release accompanying microbial Fe(II) oxidation presents a potential health concern. In contrast, production of biogenic iron sulfide minerals would not remediate the groundwater As concentration below the recommended WHO limit.
Garvicin A, a Novel Class IId Bacteriocin from Lactococcus garvieae That Inhibits Septum Formation in L. garvieae Strains
Authors: Maldonado-Barragï¿½n Antonio , Cï¿½rdenas Nivia , Martï¿½nez Beatriz , Ruiz-Barba Josï¿½ Luis , Fernï¿½ndez-Garayzï¿½bal Josï¿½ F. , Rodrï¿½guez Juan M. , Gibello Alicia
Published: May 2013

Lactococcus garvieae
21881, isolated in a human clinical case, produces a novel class IId bacteriocin, garvicin A (GarA), which is specifically active against other
L. garvieae
strains, including fish- and bovine-pathogenic isolates. Purification from active supernatants, sequence analyses, and plasmid-curing experiments identified pGL5, one of the five plasmids found in
L. garvieae
[M. Aguado-Urda et al., PLoS One 7(6):e40119, 2012], as the coding plasmid for the structural gene of GarA (
lgnA
), its putative immunity protein (
lgnI
), and the ABC transporter and its accessory protein (
lgnC
and
lgnD
). Interestingly, pGL5-cured strains were still resistant to GarA. Other putative bacteriocins encoded by the remaining plasmids were not detected during purification, pointing to GarA as the main inhibitor secreted by
L. garvieae
21881. Mode-of-action studies revealed a potent bactericidal activity of GarA. Moreover, transmission microscopy showed that GarA seems to act by inhibiting septum formation in
L. garvieae
cells. This potent and species-specific inhibition by GarA holds promise for applications in the prevention or treatment of infections caused by pathogenic strains of
L. garvieae
in both veterinary and clinical settings.
Identification of a New P335 Subgroup through Molecular Analysis of Lactococcal Phages Q33 and BM13
Authors: Mahony Jennifer , Martel Bruno , Tremblay Denise M. , Neve Horst , Heller Knut J. , Moineau Sylvain , van Sinderen Douwe
Published: May 2013

Lactococcal dairy starter strains are under constant threat from phages in dairy fermentation facilities, especially by members of the so-called 936, P335, and c2 species. Among these three phage groups, members of the P335 species are the most genetically diverse. Here, we present the complete genome sequences of two P335-type phages, Q33 and BM13, isolated in North America and representing a novel lineage within this phage group. The Q33 and BM13 genomes exhibit homology, not only to P335-type, but also to elements of the 936-type phage sequences. The two phage genomes also have close relatedness to phages infecting
Enterococcus
and
Clostridium
, a heretofore unknown feature among lactococcal P335 phages. The Q33 and BM13 genomes are organized in functionally related clusters with genes encoding functions such as DNA replication and packaging, morphogenesis, and host cell lysis. Electron micrographic analysis of the two phages highlights the presence of a baseplate more reminiscent of the baseplate of 936 phages than that of the majority of members of the P335 group, with the exception of r1t and LC3.
Pressure-Induced Endocytic Degradation of the Saccharomyces cerevisiae Low-Affinity Tryptophan Permease Tat1 Is Mediated by Rsp5 Ubiquitin Ligase and Functionally Redundant PPxY Motif Proteins
Authors: Suzuki Asaha , Mochizuki Takahiro , Uemura Satoshi , Hiraki Toshiki , Abe Fumiyoshi
Published: May 2013

Cells of
Saccharomyces cerevisiae
express two tryptophan permeases, Tat1 and Tat2, which have different characteristics in terms of their affinity for tryptophan and intracellular localization. Although the high-affinity permease Tat2 has been well documented in terms of its ubiquitin-dependent degradation, the low-affinity permease Tat1 has not yet been characterized fully. Here we show that a high hydrostatic pressure of 25 MPa triggers a degradation of Tat1 which depends on Rsp5 ubiquitin ligase and the EH domain-containing protein End3. Tat1 was resistant to a 3-h cycloheximide treatment, suggesting that it is highly stable under normal growth conditions. The ubiquitination of Tat1 most likely occurs at N-terminal lysines 29 and 31. Simultaneous substitution of arginine for the two lysines prevented Tat1 degradation, but substitution of either of them alone did not, indicating that the roles of lysines 29 and 31 are redundant. When cells were exposed to high pressure, Tat1-GFP was completely lost from the plasma membrane, while substantial amounts of Tat1
K29R-K31R
-GFP remained. The
HPG1-1
(Rsp5
P514T
) and
rsp5-ww3
mutations stabilized Tat1 under high pressure, but any one of the
rsp5-ww1
,
rsp5-ww2
, and
bul1
Δ
bul2
Δ mutations or single deletions of genes encoding arrestin-related trafficking adaptors did not. However, simultaneous loss of 9-arrestins and Bul1/Bul2 prevented Tat1 degradation at 25 MPa. The results suggest that multiple PPxY motif proteins share some essential roles in regulating Tat1 ubiquitination in response to high hydrostatic pressure.