bluuebunny/crossref_metadata_2025_split huggingface dataset:
The cytoplasmic localization of the catalytic site of CSLF6 supports a channeling model for the biosynthesis of mixed‐linkage glucan
Authors: Kim Sang‐Jin , Zemelis Starla , Keegstra Kenneth , Brandizzi Federica 
Published: December 2014

SummaryMixed‐linkage glucan (MLG) is a significant cell wall carbohydrate in grasses and an important carbon source for human consumption and biofuel production. MLG biosynthesis depends on the biochemical activity of membrane spanning glucan synthases encoded by the CSLH and CSLF cellulose synthase‐like gene families. CSLF proteins are the best characterized to date but relatively little information is known about their topology with respect to the biosynthetic membranes. In this study, we report on the topology of CSLF6 protein derived from the model grass species Brachypodium distachyon (BdCSLF6) when it is expressed in heterologous systems. Using live cell imaging and immuno‐electron microscopy analyses of tobacco epidermal cells expressing BdCSLF6, we demonstrate that a functional yellow fluorescent protein (YFP) fusion of BdCSLF6 is localized to the Golgi apparatus and that the Golgi localization of BdCSLF6 is sufficient for MLG biosynthesis. By implementing protease protection assays of BdCSLF6 expressed in the yeast Pichia pastoris, we also demonstrate that the catalytic domain, the N‐terminus and the C‐ terminus of the protein are exposed in the cytosol. Furthermore, we found that BdCSLF6 is capable of producing MLG not only in tobacco cells but also in Pichia, which generally does not produce MLG. Together, these results support the conclusion that BdCSLF6 can produce both of the linkages present in the (1,3;1,4)‐β‐d‐glucan chain of MLG and that the product is channelled at the Golgi into the secretory pathway for deposition into the cell wall.
Recombination products suggest the frequent occurrence of aberrant gene replacement in the moss Physcomitrella patens
Authors: Wendeler Edelgard , Zobell Oliver , Chrost Bozena , Reiss Bernd 
Published: December 2014

SummaryIn gene replacement, a variant of gene targeting, transformed DNA integrates into the genome by homologous recombination (HR) to replace resident sequences. Gene replacement in the moss Physcomitrella patens is extremely efficient, but often large amounts of additional DNA are integrated at the target locus. A detailed analysis of recombination junctions of PpCOL2 gene knockout mutants shows that the integrated DNA can be highly rearranged. Our data suggest that the replaced sequences were excised by HR and became integrated back into the genome by non‐homologous end‐joining (NHEJ). RAD51‐mediated strand‐invasion and subsequent strand‐exchange is central to the two‐end invasion pathway, the major gene replacement pathway in yeast. In this pathway, integration is initiated by the free ends of a single replacement vector‐derived donor molecule which then integrates as an entity. Gene replacement in P. patens is entirely RAD51‐dependent suggesting the existence of a pathway mechanistically similar to two‐end invasion. However, invasion of the two ends does not seem to be stringently coordinated in P. patens. Actually, often only one fragment end became integrated by HR, or one‐sided integration of two independent donor fragments occurred simultaneously leading to a double‐strand break that is subsequently sealed by NHEJ and thus causes the observed rearrangements.
Long‐term non‐invasive and continuous measurements of legume nodule activity
Authors: Cabeza Ricardo A. , Liese Rebecca , Fischinger Stephanie A. , Sulieman Saad , Avenhaus Ulrike , Lingner Annika , Hein Hans , Koester Beke , Baumgarten Vanessa , Dittert Klaus , Schulze Joachim 
Published: December 2014

SummarySymbiotic nitrogen fixation is a process of considerable economic, ecological and scientific interest. The central enzyme nitrogenase reduces H+ alongside N2, and the evolving H2 allows a continuous and non‐invasive in vivo measurement of nitrogenase activity. The objective of this study was to show that an elaborated set‐up providing such measurements for periods as long as several weeks will produce specific insight into the nodule activity's dependence on environmental conditions and genotype features. A system was developed that allows the air‐proof separation of a root/nodule and a shoot compartment. H2 evolution in the root/nodule compartment can be monitored continuously. Nutrient solution composition, temperature, CO2 concentration and humidity around the shoots can concomitantly be maintained and manipulated. Medicago truncatula plants showed vigorous growth in the system when relying on nitrogen fixation. The set‐up was able to provide specific insights into nitrogen fixation. For example, nodule activity depended on the temperature in their surroundings, but not on temperature or light around shoots. Increased temperature around the nodules was able to induce higher nodule activity in darkness versus light around shoots for a period of as long as 8 h. Conditions that affected the N demand of the shoots (ammonium application, Mg or P depletion, super numeric nodules) induced consistent and complex daily rhythms in nodule activity. It was shown that long‐term continuous measurements of nodule activity could be useful for revealing special features in mutants and could be of importance when synchronizing nodule harvests for complex analysis of their metabolic status.
Tagging Strategies Strongly Affect the Fate of Overexpressed Caveolin‐1
Authors: Han Bing , Tiwari Ajit , Kenworthy Anne K. 
Published: December 2014

AbstractCaveolin‐1 (Cav1) is the primary scaffolding protein of caveolae, flask‐shaped invaginations of the plasma membrane thought to function in endocytosis, mechanotransduction, signaling and lipid homeostasis. A significant amount of our current knowledge about caveolins and caveolae is derived from studies of transiently overexpressed, C‐terminally tagged caveolin proteins. However, how different tags affect the behavior of ectopically expressed Cav1 is still largely unknown. To address this question, we performed a comparative analysis of the subcellular distribution, oligomerization state and detergent resistance of transiently overexpressed Cav1 labeled with three different C‐terminal tags (EGFP, mCherry and myc). We show that addition of fluorescent protein tags enhances the aggregation and/or degradation of both wild‐type Cav1 and an oligomerization defective P132L mutant. Strikingly, complexes formed by overexpressed Cav1 fusion proteins excluded endogenous Cav1 and Cav2, and the properties of native caveolins were largely preserved even when abnormal aggregates were present in cells. These findings suggest that differences in tagging strategies may be a source of variation in previously published studies of Cav1 and that overexpressed Cav1 may exert functional effects outside of caveolae. They also highlight the need for a critical re‐evaluation of current knowledge based on transient overexpression of tagged Cav1.
Evaluation of Dutch guideline for just‐in‐time addition of plerixafor to stem cell mobilization in patients who fail with granulocyte–colony‐stimulating factor
Authors: Bilgin Yavuz M. , Visser Otto , Beckers Erik A.M. , te Boome Liane CJ , Huisman Cynthia , Ypma Paula F. , Croockewit Alexandra J. , Netelenbos Tanja , Kramer Ellen PA , de Greef Georgine E. 
Published: December 2014

BackgroundPlerixafor in combination with granulocyte‐colony‐stimulating factor (G‐CSF) is approved for the use of stem cell collection in patients who fail to mobilize on G‐CSF. In 2009 the Stem Cell Working Party of the Dutch‐Belgian Cooperative Trial group for Hematology Oncology (HOVON) composed a guideline for the use of plerixafor. According to this guideline it is recommended to add plerixafor to G‐CSF in patients with circulating CD34+ cell counts of fewer than 20 × 106/L on 2 consecutive days accompanied by increasing white blood cells.Study Design and MethodsIn this analysis we evaluated retrospectively the outcome of the use of this guideline in the Netherlands. In total 111 patients received plerixafor with a median one administration (range, one to four administrations). Of these patients 55.8% had non‐Hodgkin lymphoma, 31.5% multiple myeloma, 8.1% Hodgkin lymphoma, and 4.5% nonhematologic malignancies.ResultsIn 63.9% patients sufficient numbers of CD34+ cells were collected. In patients with multiple myeloma more successful mobilizations with plerixafor were observed compared to patients with non‐Hodgkin lymphoma (71.4% vs. 61.3%). In patients with circulating CD34+ cell counts of at least 2.0 × 106/L before administration of plerixafor a successful mobilization was achieved in 76.5%, and in the patients with very low (0‐1 × 106/L) circulating CD34+ cell counts the success rate was 44.2%.ConclusionApplication of the HOVON guideline on the just‐in‐time administration of plerixafor is effective for mobilization of hematopoietic stem cells in the majority of patients. Stem cell yield in patients with non‐Hodgkin lymphoma was lower compared to patients with multiple myeloma. Also patients with very low circulating CD34+ cells before addition of plerixafor might benefit from this approach.
Red blood cell Thomsen‐Friedenreich antigen expression and galectin‐3 plasma concentrations in Streptococcus pneumoniae–associated hemolytic uremic syndrome and hemolytic anemia
Authors: Burin des Roziers Nicolas , Chadebech Philippe , Bodivit Gwellaouen , Guinchard Emmanuelle , Bruneel Arnaud , Dupré Thierry , Chevret Laurent , Jugie Myriam , Gallon Philippe , Bierling Philippe , Noizat‐Pirenne France 
Published: December 2014

BackgroundPneumococcal hemolytic uremic syndrome (P‐HUS) is a rare but severe complication of invasive pneumococcal disease (IPD) in young children. Consensual biologic diagnosis criteria are currently lacking.Study Design and MethodsA prospective study was conducted on 10 children with culture‐confirmed IPD. Five presented with full‐blown P‐HUS, three had an incomplete form with hemolytic anemia and mild or no uremia (P‐HA), and two had neither HUS nor HA. Thomsen‐Friedenreich (T), Th, and Tk cryptantigens and sialic acid expression were determined on red blood cells (RBCs) with peanut (PNA), Glycine soja (SBA), Bandeiraea simplicifolia II, and Maackia amurensis lectins. Plasma concentrations of the major endogenous T‐antigen–binding protein, galectin‐3 (Gal‐3), were analyzed.ResultsWe found that RBCs strongly reacted with PNA and SBA lectins in all P‐HUS and P‐HA patients. Three P‐HUS and three P‐HA patients showed also concomitant Tk activation. Direct antiglobulin test (DAT) was positive in three P‐HUS (one with anti‐C3d and two with anti‐IgG) and two P‐HA patients (one with anti‐C3d and one with anti‐IgG). RBCs derived from the two uncomplicated IPD patients reacted with PNA but not with SBA lectin. Gal‐3 plasma concentrations were increased in all P‐HUS patients.ConclusionsThe results indicate high levels of neuraminidase activity and desialylation in both P‐HUS and P‐HA patients. T‐antigen activation is more sensitive than DAT for P‐HUS diagnosis. Combining PNA and SBA lectins is needed to improve the specificity of T‐antigen activation. High concentrations of Gal‐3 in P‐HUS patients suggest that Gal‐3 may contribute to the pathogenesis of P‐HUS.
Evaluation of the appropriateness of frozen plasma usage after introduction of prothrombin complex concentrates: a retrospective study
Authors: Shih A. W. , Kolesar E. , Ning S. , Manning N. , Arnold D. M. , Crowther M. A. 
Published: December 2014

BackgroundProthrombin complex concentrates (PCCs) can be used instead of frozen plasma (FP) transfusion to reverse the effect of warfarin. Audits have demonstrated over usage of FP transfusions even before the introduction of PCC. The objective of this study was to determine the appropriateness of current FP transfusion practice in the current era since the introduction of PCCs.MethodsA retrospective cohort study of consecutive patients receiving FP over 3 months was carried out. Each episode of FP use over a 24‐h period was adjudicated independently by two reviewers as appropriate (consistent with Canadian/AABB guidelines), appropriate but inconsistent with guidelines or inappropriate. Discrepancies were resolved by a third reviewer. Use of FP to reverse warfarin was considered inappropriate. FP usage from previous years was assessed as baseline.ResultsDuring the study period, 111 FP transfusions were administered. 74·8% of FP usage occurred in the ICU. The proportion of FP transfusions that were deemed appropriate, inconsistent yet appropriate or inappropriate were 33/89 (37·1%), 16/89 (18·0%) and 40/89 (44·9%), respectively, when use of FP for therapeutic plasma exchange was excluded. The most common reasons for inappropriate use were the absence of bleeding with an increased INR or warfarin reversal.ConclusionOur study is the first to audit FP transfusions in the post‐PCC era in Canada. FP usage remains inappropriately high in INR prolongation without another indication or to reverse warfarin. Targeted interventions to reduce FP usage in the future should focus on the ICU and on education about warfarin reversal.
Wnt signalling suppresses voltage‐dependent Na+ channel expression in postnatal rat cardiomyocytes
Authors: Liang Wenbin , Cho Hee Cheol , Marbán Eduardo 
Published: December 2014

Key points
Wnt signalling is activated in arrhythmogenic heart diseases, but its role in the regulation of cardiac ion channel expression is unknown.
Exposure of neonatal rat ventricular myocytes to Wnt3a, an activator of canonical Wnt signalling, decreases Scn5a mRNA, Nav1.5 protein and Na+ current density.
Wnt3a does not affect the inward rectifier K+ current or L‐type Ca2+ channels.
The Wnt pathway is a negative regulator of cardiac Na+ channel expression and may play a role in altered ion channel expression in heart disease.
AbstractWnt signalling plays crucial roles in heart development, but is normally suppressed postnatally. In arrhythmogenic conditions, such as cardiac hypertrophy and heart failure, Wnt signalling is reactivated. To explore the potential role of Wnt signalling in arrhythmogenic electrical remodelling, we examined voltage‐dependent ion channels in cardiomyocytes. Treatment of neonatal rat ventricular myocytes with either recombinant Wnt3a protein or CHIR‐99021 (CHIR, a glycogen synthase kinase‐3β inhibitor) caused a dose‐dependent increase in Wnt target gene expression (Axin2 and Lef1), indicating activation of the Wnt/β‐catenin pathway. Cardiac Na+ current (INa) density was reduced by Wnt3a (−20 ± 4 vs. control −59 ± 7 pA pF−1, at −30 mV) or CHIR (−22 ± 5 pA pF−1), without changes in steady‐state activation, inactivation or repriming kinetics. Wnt3a and CHIR also produced dose‐dependent reductions in the mRNA level of Scn5a (the cardiac Na+ channel α subunit gene), as well as a 56% reduction (by Wnt3a) in the Nav1.5 protein level. Consistent with INa reduction, action potentials in Wnt3a‐treated neonatal rat ventricular myocytes had a lower upstroke amplitude (91 ± 3 vs. control 137 ± 2 mV) and decreased maximum upstroke velocity (70 ± 10 vs. control 163 ± 15 V s−1). In contrast, inward rectifier K+ current and L‐type Ca2+ channels were not affected by Wnt3a treatment. Taken together, our data indicate that the Wnt/β‐catenin pathway suppresses INa in postnatal cardiomyocytes and may contribute to ion channel remodelling in heart disease.
The Experimental Study of Matching Between Centrifugal Compressor Impeller and Diffuser
Authors: Tamaki Hideaki , Nakao Hidefumi , Saito Masayasu 
Published: December 2014

The centrifugal compressor for a marine use turbocharger with its design pressure ratio of 3.2 was tested with a vaneless diffuser and various vaned diffusers. Vaned diffusers were chosen to cover impeller operating range as broad as possible. The analysis of the static pressure ratio in the impeller and the diffusing system — consists of the diffuser and scroll — showed that there were four possible combinations of characteristics of impeller pressure ratio and that of diffusing system pressure ratio.
The flow rate, QP, where the impeller achieved maximum static pressure ratio was the critical flow rate determined surge flow rate of the centrifugal compressor. In order to operate the compressor less than QP, the diffusing system whose pressure recovery factor was steep negative slope near QP was needed. Using the diffuser throat area less than a certain value, the compressor efficiency deteriorated, however the compressor stage pressure ratio was almost constant. In this study, the compressor by reducing the diffuser throat area could be operated at the flow rate less than 40% of its design flow rate. Analysis of the pressure ratio in the impeller and diffusing systems at design and off design speeds showed that the irregularities in surge line occurred when the component which controlled the negative slope on the compressor stage pressure ratio changed.
3-D Conjugate Flow and Heat Transfer Investigations of a Film-Cooled Turbine Guide Vane With Different Blowing Ratios
Authors: Bohn Dieter E. , Heuer Tom , Kusterer Karsten A. 
Published: December 2014

Film-cooling has become a widely used cooling method in present day gas turbines. Cooling gas ejection at the leading edge serves to protect the entire vane surface from contact with the hot gas. With doing this, material temperatures are reduced in order to guarantee an economically acceptable life span of the vane.
This paper describes the application of a numerical method for the conjugate calculation of internal and external fluid flows and the heat transfer in and through the blade walls of a film-cooled turbine guide vane. The advantage of this approach is that it is possible to predict fluid flow properties and wall temperatures without the need for additional heat transfer conditions or temperature conditions at the external surfaces of the vane. This is a great advantage because the desired data are either unknown or not available for the calculation in the design process of new cooled blades or vanes. In a complete calculation of external and internal flows, no additional boundary conditions at the internal surfaces of the cooling geometry are needed either. Another advantage is the interaction of fluid flow and heat transfer which is taken into account by the conjugate calculation.
In the 3-D numerical experiment to be presented, the influence of leading edge cooling fluid ejection on the temperature distribution in the vane material is investigated. The cooling fluid is ejected through two slots at the leading edge. The calculations are performed for three blowing ratios in order to investigate the efficiency of the cooling method. Realistic temperature ratios of cooling-fluid flow and main flow are considered. Such information is very useful in the aero thermal design process of new cooling configurations, since the amount of experimental work can be minimized. The results show the influence of complex 3-D flow phenomena (e.g. passage vortex) on the cooling fluid distribution on the vane surface as a function of the chosen blowing factor. Due to the influence of the passage vortex, the cooling fluid is displaced and leaves the vane surface near the side-wall uncovered against the hot gas. Furthermore, cooling fluid displacement on the pressure side according to the ejection slot geometry leads to another unprotected region on the vane surface. These effects have severe consequences on the thermal load of the vane and can reduce its life span.
University Expertise and the Oil and Gas Industry: Development of Cost Effective Solutions to Applied Oil-Spill-Related Research Issues
Authors: Davis Donald W. , Guidry Roland J. 
Published: December 2014

Immediately after the Exxon Valdez incident, the United States Oil Pollution Act of 1990 was passed. This Act clarified the lines of responsibility associated with future oil spills. In addition to this Federal legislation, Louisiana lawmakers in 1991 enacted the Oil Spill Prevention and Response Act. Financial awards associated with this Act support a wide-range of research activities. Since 1993, 24 projects have been funded. The scope and nature of this research includes:
• Oil Spill Awareness through Geoscience Education (OSAGE);
• Used Oil Recycling in Louisiana’s Coastal Communities;
• Evaluation and Characterization of Sorbents;
• Landsat TM and Synthetic Aperture Radar to Facilitate Coastline Delineation;
• Environmental Effects and Effectiveness of In-Situ Burning in Wetlands;
• Bioremediation Protocol for Small-Scale Oil Spills;
• Oil Spill Risk on Louisiana’s Largest Waterway;
• River Time-of-Travel Modeling;
• Composting Technology for Practical and Safe Remediation of Oil-Spill Residuals;
• Predictability of Oceanic and Atmospheric Conditions off the Mississippi Delta; and
• Phytoremediation for Oil Spill Cleanup and Habitat Restoration in Louisiana’s Marshes.
Each of these projects, and others, are the result of the marriage of industry and university researchers in the identification and solution of applied oil-spill-related problems. The alliance is a good one. Important environmental issues are addressed because the selection process ensures each research initiative has the potential of being implemented by the response community. The work and knowledge gained from these projects is a clear indication of how industry and the university community can function in a collaborative manner to solve important issues — a significant partnership that clearly shows how both can benefit and a model for others to follow.
A Better Way to Support Horizontal Pressure Vessels Subject to Thermal Loading
Authors: Tooth Alwyn S. , Cheung John S. T. , Ng Heong W. , Ong Lin S. , Nadarajah Chithranjan 
Published: December 2014

When storing liquids at high temperature, in horizontal vessels, the current design methods aim to minimise the thermal stresses by introducing a sliding surface at the base of one of the twin saddle supports. However, regular site maintenance is required to ensure that adequate sliding is achieved This may be difficult and costly to carry out.
The aim of the present work, therefore, is to dispense with the sliding support and to provide saddle designs which although fixed to the platform, or foundation, do not result in the storage/pressure vessel being over-stressed when thermal loading occurs. The paper provides general recommendations for the most appropriate saddle geometries, and details the way in which ‘Design by Analysis’ and ‘Fatigue Life Assessments’ may be carried out using the stresses which arise from these designs.
Nondestructive Evaluation (NDE): Auralization of Ultrasonic Signals for the Detection of Corrosion
Authors: Holt Amos E. , Light Glenn M. 
Published: December 2014

During nondestructive evaluation (NDE), an instrument is used to receive inspection signals and display them for visual interpretation. Southwest Research Institute (SwRI) has developed technology that allows NDE signals generated from conventional ultrasonic instrumentation to be completely converted into audible (aural) signals to enhance reliability of evaluation through the use of two senses (ears and eyes) to improve inspection reliability.
For example, detection of hidden corrosion on aircraft structures using conventional ultrasonic testing (UT) techniques is difficult because of thin walls and corrosion topography. These characteristics require use of high-frequency, high-spatial-resolution transducers to attempt to detect backwall signal amplitude. It is difficult to visually discriminate backwall signals of thin wing structure from the normal ringdown of the transducer.
Using aural UT, a trained inspector can listen to sounds generated by aural UT equipment and detect the presence of hidden corrosion with higher reliability than using conventional UT.