Further characterization of ADAMTS-13 inactivation by thrombin

Background: The multimeric size and platelet-tethering function of von Willebrand factor (VWF) are modulated by the plasma metalloprotease, a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS-13). In vitro ADAMTS-13 is susceptible to proteolytic inactivation by thrombin. Objectives: In this study, we aimed to characterize the inactivation of ADAMTS-13 by thrombin and to assess its physiological significance. Methods and results: By N-terminal sequencing of cleavage products, and by mutagenesis, we identified the principal thrombin cleavage sites in ADAMTS-13 as R257 and R1176. Using a library of 76 thrombin mutants, we highlighted the functional importance of exosite I on thrombin in the proteolysis of ADAMTS-13. Proteolysis of ADAMTS-13 by thrombin caused an 8-fold reduction in its affinity for VWF that contributed to its loss of VWF-cleaving function. Intriguingly, thrombin-cleaved ADAMTS-13 both bound and proteolyzed a short recombinant VWF A2 domain substrate (VWF115) normally. Following activation of coagulation in normal plasma, endogenous ADAMTS-13, but not added ADAMTS-13, appeared resistant to coagulation-induced fragmentation. An estimation of the Km for ADAMTS-13 proteolysis by thrombin was appreciably higher than the physiological concentration of ADAMTS-13. This was corroborated by the comparatively low affinity of ADAMTS-13 for thrombin (KD 95 nm). Conclusions: Together, our data suggest that ADAMTS-13 is protected from rapid proteolytic inactivation by thrombin in normal plasma. Whether this remains the case under pathological situations involving elevated/sustained generation of thrombin remains unclear

Frequent burning promotes invasions of alien plants into a mesic African savanna

Fire is both inevitable and necessary for maintaining the structure and functioning of mesic savannas. Without disturbances such as fire and herbivory, tree cover can increase at the expense of grass cover and over time dominate mesic savannas. Consequently, repeated burning is widely used to suppress tree recruitment and control bush encroachment. However, the effect of regular burning on invasion by alien plant species is little understood. Here, vegetation data from a long-term fire experiment, which began in 1953 in a mesic Zimbabwean savanna, were used to test whether the frequency of burning promoted alien plant invasion. The fire treatments consisted of late season fires, lit at 1-, 2-, 3-, and 4-year intervals, and these regularly burnt plots were compared with unburnt plots. Results show that over half a century of frequent burning promoted the invasion by alien plants relative to areas where fire was excluded. More alien plant species became established in plots that had a higher frequency of burning. The proportion of alien species in the species assemblage was highest in the annually burnt plots followed by plots burnt biennially. Alien plant invasion was lowest in plots protected from fire but did not differ significantly between plots burnt triennially and quadrennially. Further, the abundance of five alien forbs increased significantly as the interval (in years) between fires became shorter. On average, the density of these alien forbs in annually burnt plots was at least ten times as high as the density of unburnt plots. Plant diversity was also altered by long-term burning. Total plant species richness was significantly lower in the unburnt plots compared to regularly burnt plots. These findings suggest that frequent burning of mesic savannas enhances invasion by alien plants, with short intervals between fires favouring alien forbs. Therefore, reducing the frequency of burning may be a key to minimising the risk of alien plant spread into mesic savannas, which is important because invasive plants pose a threat to native biodiversity and may alter savanna functioning

‘Trying to pin down jelly’ - exploring intuitive processes in quality assessment for meta-ethnography

Background: Studies that systematically search for and synthesise qualitative research are becoming more evident in health care, and they can make an important contribution to patient care. However, there is still no agreement as to whether, or how we should appraise studies for inclusion. We aimed to explore the intuitive processes that determined the ‘quality’ of qualitative research for inclusion in qualitative research syntheses. We were particularly interested to explore the way that knowledge was constructed. Methods: We used qualitative methods to explore the process of quality appraisal within a team of seven qualitative researchers funded to undertake a meta-ethnography of chronic non-malignant musculoskeletal pain. Team discussions took place monthly between October 2010 and June 2012 and were recorded and transcribed. Data was coded and organised using constant comparative method. The development of our conceptual analysis was both iterative and collaborative. The strength of this team approach to quality came from open and honest discussion, where team members felt free to agree, disagree, or change their position within the safety of the group. Results: We suggest two core facets of quality for inclusion in meta-ethnography - (1) Conceptual clarity; how clearly has the author articulated a concept that facilitates theoretical insight. (2) Interpretive rigour; fundamentally, can the interpretation ‘be trusted?’ Our findings showed that three important categories help the reader to judge interpretive rigour: (ii) What is the context of the interpretation? (ii) How inductive is the interpretation? (iii) Has the researcher challenged their interpretation? Conclusions: We highlight that methods alone do not determine the quality of research for inclusion into a meta-ethnography. The strength of a concept and its capacity to facilitate theoretical insight is integral to meta-ethnography, and arguably to the quality of research. However, we suggest that to be judged ‘good enough’ there also needs to be some assurance that qualitative findings are more than simply anecdotal. Although our conceptual model was developed specifically for meta-ethnography, it may be transferable to other research methodologies

Do Interventions Designed to Support Shared Decision-Making Reduce Health Inequalities? : A Systematic Review and Meta-Analysis

Copyright: © 2014 Durand et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Background: Increasing patient engagement in healthcare has become a health policy priority. However, there has been concern that promoting supported shared decision-making could increase health inequalities. Objective: To evaluate the impact of SDM interventions on disadvantaged groups and health inequalities. Design: Systematic review and meta-analysis of randomised controlled trials and observational studies.Peer reviewe

Meta-analyses and Forest plots using a microsoft excel spreadsheet: step-by-step guide focusing on descriptive data analysis

<p>Abstract</p> <p>Background</p> <p>Meta-analyses are necessary to synthesize data obtained from primary research, and in many situations reviews of observational studies are the only available alternative. General purpose statistical packages can meta-analyze data, but usually require external macros or coding. Commercial specialist software is available, but may be expensive and focused in a particular type of primary data. Most available softwares have limitations in dealing with descriptive data, and the graphical display of summary statistics such as incidence and prevalence is unsatisfactory. Analyses can be conducted using Microsoft Excel, but there was no previous guide available.</p> <p>Findings</p> <p>We constructed a step-by-step guide to perform a meta-analysis in a Microsoft Excel spreadsheet, using either fixed-effect or random-effects models. We have also developed a second spreadsheet capable of producing customized forest plots.</p> <p>Conclusions</p> <p>It is possible to conduct a meta-analysis using only Microsoft Excel. More important, to our knowledge this is the first description of a method for producing a statistically adequate but graphically appealing forest plot summarizing descriptive data, using widely available software.</p

Maximising response to postal questionnaires – A systematic review of randomised trials in health research

Background Postal self-completion questionnaires offer one of the least expensive modes of collecting patient based outcomes in health care research. The purpose of this review is to assess the efficacy of methods of increasing response to postal questionnaires in health care studies on patient populations. Methods The following databases were searched: Medline, Embase, CENTRAL, CDSR, PsycINFO, NRR and ZETOC. Reference lists of relevant reviews and relevant journals were hand searched. Inclusion criteria were randomised trials of strategies to improve questionnaire response in health care research on patient populations. Response rate was defined as the percentage of questionnaires returned after all follow-up efforts. Study quality was assessed by two independent reviewers. The Mantel-Haenszel method was used to calculate the pooled odds ratios. Results Thirteen studies reporting fifteen trials were included. Implementation of reminder letters and telephone contact had the most significant effect on response rates (odds ratio 3.7, 95% confidence interval 2.30 to 5.97 p = <0.00001). Shorter questionnaires also improved response rates to a lesser degree (odds ratio 1.4, 95% confidence interval 1.19 to 1.54). No evidence was found that incentives, re-ordering of questions or including an information brochure with the questionnaire confer any additional advantage. Conclusion Implementing repeat mailing strategies and/or telephone reminders may improve response to postal questionnaires in health care research. Making the questionnaire shorter may also improve response rates. There is a lack of evidence to suggest that incentives are useful. In the context of health care research all strategies to improve response to postal questionnaires require further evaluation

A meta-analytic review of stand-alone interventions to improve body image

Objective Numerous stand-alone interventions to improve body image have been developed. The present review used meta-analysis to estimate the effectiveness of such interventions, and to identify the specific change techniques that lead to improvement in body image. Methods The inclusion criteria were that (a) the intervention was stand-alone (i.e., solely focused on improving body image), (b) a control group was used, (c) participants were randomly assigned to conditions, and (d) at least one pretest and one posttest measure of body image was taken. Effect sizes were meta-analysed and moderator analyses were conducted. A taxonomy of 48 change techniques used in interventions targeted at body image was developed; all interventions were coded using this taxonomy. Results The literature search identified 62 tests of interventions (N = 3,846). Interventions produced a small-to-medium improvement in body image (d+ = 0.38), a small-to-medium reduction in beauty ideal internalisation (d+ = -0.37), and a large reduction in social comparison tendencies (d+ = -0.72). However, the effect size for body image was inflated by bias both within and across studies, and was reliable but of small magnitude once corrections for bias were applied. Effect sizes for the other outcomes were no longer reliable once corrections for bias were applied. Several features of the sample, intervention, and methodology moderated intervention effects. Twelve change techniques were associated with improvements in body image, and three techniques were contra-indicated. Conclusions The findings show that interventions engender only small improvements in body image, and underline the need for large-scale, high-quality trials in this area. The review identifies effective techniques that could be deployed in future interventions

Fundamental spray and combustion measurements of soy methyl-ester biodiesel

Although biodiesel has begun to penetrate the fuel market, its effect on injection processes, combustion and emission formation under diesel engine conditions remains somewhat unclear. Typical exhaust measurements from engines running biodiesel indicate that particulate matter, carbon monoxide and unburnt hydrocarbons are decreased, whereas nitrogen oxide emissions tend to be increased. However, these observations are the result of complex interactions between physical and chemical processes occurring in the combustion chamber, for which understanding is still needed. To characterize and decouple the physical and chemical influences of biodiesel on spray mixing, ignition, combustion and soot formation, a soy methyl-ester (SME) biodiesel is injected into a constant-volume combustion facility under diesel-like operating conditions. A range of optical diagnostics is performed, comparing biodiesel to a conventional #2 diesel at the same injection and ambient conditions. Schlieren high-speed imaging shows virtually the same vapour-phase penetration for the two fuels, while simultaneous Mie-scatter imaging shows that the maximum liquid-phase penetration of biodiesel is higher than diesel. Differences in the liquid-phase penetration are expected because of the different boiling-point temperatures of the two fuels. However, the different liquid-phase penetration does not affect overall mixing rate and downstream vapour-phase penetration because each fuel spray has similar momentum and spreading angle. For the biodiesel and diesel samples used in this study, the ignition delay and lift-off length are only slightly less for biodiesel compared to diesel, consistent with the fuel cetane number (51 for biodiesel, 46 for diesel). Because of the similarity in lift-off length, the differences in equivalence ratio distribution at the lift-off length are mainly affected by the oxygen content of the fuels. For biodiesel, the equivalence ratio is reduced, which, along with the fuel molecular structure and oxygen content, significantly affects soot formation downstream. Spatially resolved soot volume fraction measurements obtained by combining line-of-sight laser extinction measurements with planar laser-induced incandescence imaging show that the soot concentration can be reduced by an order of magnitude for biodiesel. These integrated measurements of spray mixing, combustion and quantitative soot concentration provide new validation data for the development of computational fluid dynamics spray, combustion and soot formation models suitable for the latest biofuels.This work was supported by the Spanish Ministry of Science and Innovation for Jean-Guillaume Nerva's visiting research, through the OPTICOMB project [TRA2007-67961-C03-01].Nerva, J.; Genzale, CL.; Kook, S.; García Oliver, JM.; Pickett, LM. (2013). Fundamental spray and combustion measurements of soy methyl-ester biodiesel. International Journal of Engine Research. 14(4):373-390. https://doi.org/10.1177/1468087412456688S373390144(2009). World Energy Outlook 2009. World Energy Outlook. doi:10.1787/weo-2009-enMonyem, A., & H. Van Gerpen, J. (2001). The effect of biodiesel oxidation on engine performance and emissions. Biomass and Bioenergy, 20(4), 317-325. doi:10.1016/s0961-9534(00)00095-7LAPUERTA, M., ARMAS, O., & RODRIGUEZFERNANDEZ, J. (2008). Effect of biodiesel fuels on diesel engine emissions. Progress in Energy and Combustion Science, 34(2), 198-223. doi:10.1016/j.pecs.2007.07.001Fisher, B. T., Knothe, G., & Mueller, C. J. (2010). Liquid-Phase Penetration under Unsteady In-Cylinder Conditions: Soy- and Cuphea-Derived Biodiesel Fuels Versus Conventional Diesel. Energy & Fuels, 24(9), 5163-5180. doi:10.1021/ef100594pFang, T., Lin, Y.-C., Foong, T. M., & Lee, C. (2009). Biodiesel combustion in an optical HSDI diesel engine under low load premixed combustion conditions. Fuel, 88(11), 2154-2162. doi:10.1016/j.fuel.2009.02.033Pastor, J. V., García-Oliver, J. M., Nerva, J.-G., & Giménez, B. (2011). Fuel effect on the liquid-phase penetration of an evaporating spray under transient diesel-like conditions. Fuel, 90(11), 3369-3381. doi:10.1016/j.fuel.2011.05.006Fisher, B. T., & Mueller, C. J. (2010). Liquid penetration length of heptamethylnonane and trimethylpentane under unsteady in-cylinder conditions. Fuel, 89(10), 2673-2696. doi:10.1016/j.fuel.2010.04.024Kim, H. J., Park, S. H., Suh, H. K., & Lee, C. S. (2009). Atomization and Evaporation Characteristics of Biodiesel and Dimethyl Ether Compared to Diesel Fuel in a High-Pressure Injection System. Energy & Fuels, 23(3), 1734-1742. doi:10.1021/ef800811gSuh, H. K., Roh, H. G., & Lee, C. S. (2008). Spray and Combustion Characteristics of Biodiesel∕Diesel Blended Fuel in a Direct Injection Common-Rail Diesel Engine. Journal of Engineering for Gas Turbines and Power, 130(3). doi:10.1115/1.2835354Pickett, L. M., & Siebers, D. L. (2006). Soot Formation in Diesel Fuel Jets Near the Lift-Off Length. International Journal of Engine Research, 7(2), 103-130. doi:10.1243/146808705x57793Pickett, L. M., Kook, S., Persson, H., & Andersson, Ö. (2009). Diesel fuel jet lift-off stabilization in the presence of laser-induced plasma ignition. Proceedings of the Combustion Institute, 32(2), 2793-2800. doi:10.1016/j.proci.2008.06.082Yoo, C. S., Richardson, E. S., Sankaran, R., & Chen, J. H. (2011). A DNS study on the stabilization mechanism of a turbulent lifted ethylene jet flame in highly-heated coflow. Proceedings of the Combustion Institute, 33(1), 1619-1627. doi:10.1016/j.proci.2010.06.147Pastor, J. V., Payri, R., Gimeno, J., & Nerva, J. G. (2009). Experimental Study on RME Blends: Liquid-Phase Fuel Penetration, Chemiluminescence, and Soot Luminosity in Diesel-Like Conditions. Energy & Fuels, 23(12), 5899-5915. doi:10.1021/ef9007328Benajes, J., Molina, S., Novella, R., & Amorim, R. (2010). Study on Low Temperature Combustion for Light-Duty Diesel Engines. Energy & Fuels, 24(1), 355-364. doi:10.1021/ef900832cPickett, L. M., & Siebers, D. L. (2002). An investigation of diesel soot formation processes using micro-orifices. Proceedings of the Combustion Institute, 29(1), 655-662. doi:10.1016/s1540-7489(02)80084-0Siebers, D. L., & Pickett, L. M. (2004). Injection Pressure and Orifice Diameter Effects on Soot in DI Diesel Fuel Jets. Thermo- and Fluid Dynamic Processes in Diesel Engines 2, 109-132. doi:10.1007/978-3-662-10502-3_7Pickett, L. M., & Siebers, D. L. (2004). Soot in diesel fuel jets: effects of ambient temperature, ambient density, and injection pressure. Combustion and Flame, 138(1-2), 114-135. doi:10.1016/j.combustflame.2004.04.006Cheng, A. S., Upatnieks, A., & Mueller, C. J. (2006). Investigation of the impact of biodiesel fuelling on NOx emissions using an optical direct injection diesel engine. International Journal of Engine Research, 7(4), 297-318. doi:10.1243/14680874jer05005Cheng, A. S. (Ed), Upatnieks, A., & Mueller, C. J. (2007). Investigation of Fuel Effects on Dilute, Mixing-Controlled Combustion in an Optical Direct-Injection Diesel Engine. Energy & Fuels, 21(4), 1989-2002. doi:10.1021/ef0606456Klein-Douwel, R. J. H., Donkerbroek, A. J., van Vliet, A. P., Boot, M. D., Somers, L. M. T., Baert, R. S. G., … ter Meulen, J. J. (2009). Soot and chemiluminescence in diesel combustion of bio-derived, oxygenated and reference fuels. Proceedings of the Combustion Institute, 32(2), 2817-2825. doi:10.1016/j.proci.2008.06.140Fang, T., & Lee, C. F. (2009). Bio-diesel effects on combustion processes in an HSDI diesel engine using advanced injection strategies. Proceedings of the Combustion Institute, 32(2), 2785-2792. doi:10.1016/j.proci.2008.07.031Payri, F., Pastor, J. V., Nerva, J.-G., & Garcia-Oliver, J. M. (2011). Lift-Off Length and KL Extinction Measurements of Biodiesel and Fischer-Tropsch Fuels under Quasi-Steady Diesel Engine Conditions. SAE International Journal of Engines, 4(2), 2278-2297. doi:10.4271/2011-24-0037Kook, S., & Pickett, L. M. (2012). Liquid length and vapor penetration of conventional, Fischer–Tropsch, coal-derived, and surrogate fuel sprays at high-temperature and high-pressure ambient conditions. Fuel, 93, 539-548. doi:10.1016/j.fuel.2011.10.004Settles, G. S. (2001). Schlieren and Shadowgraph Techniques. doi:10.1007/978-3-642-56640-0Pickett, L. M., Manin, J., Genzale, C. L., Siebers, D. L., Musculus, M. P. B., & Idicheria, C. A. (2011). Relationship Between Diesel Fuel Spray Vapor Penetration/Dispersion and Local Fuel Mixture Fraction. SAE International Journal of Engines, 4(1), 764-799. doi:10.4271/2011-01-0686MUSCULUS, M., & PICKETT, L. (2005). Diagnostic considerations for optical laser-extinction measurements of soot in high-pressure transient combustion environments. Combustion and Flame, 141(4), 371-391. doi:10.1016/j.combustflame.2005.01.013Williams, T. C., Shaddix, C. R., Jensen, K. A., & Suo-Anttila, J. M. (2007). Measurement of the dimensionless extinction coefficient of soot within laminar diffusion flames. International Journal of Heat and Mass Transfer, 50(7-8), 1616-1630. doi:10.1016/j.ijheatmasstransfer.2006.08.024Kook, S., & Pickett, L. M. (2011). Soot volume fraction and morphology of conventional and surrogate jet fuel sprays at 1000-K and 6.7-MPa ambient conditions. Proceedings of the Combustion Institute, 33(2), 2911-2918. doi:10.1016/j.proci.2010.05.073De Francqueville, L., Bruneaux, G., & Thirouard, B. (2010). Soot Volume Fraction Measurements in a Gasoline Direct Injection Engine by Combined Laser Induced Incandescence and Laser Extinction Method. SAE International Journal of Engines, 3(1), 163-182. doi:10.4271/2010-01-0346Musculus, M. P. B., & Kattke, K. (2009). Entrainment Waves in Diesel Jets. SAE International Journal of Engines, 2(1), 1170-1193. doi:10.4271/2009-01-1355Desantes, J. M., Pastor, J. V., García-Oliver, J. M., & Pastor, J. M. (2009). A 1D model for the description of mixing-controlled reacting diesel sprays. Combustion and Flame, 156(1), 234-249. doi:10.1016/j.combustflame.2008.10.008Idicheria, C. A., & Pickett, L. M. (2011). Ignition, soot formation, and end-of-combustion transients in diesel combustion under high-EGR conditions. International Journal of Engine Research, 12(4), 376-392. doi:10.1177/1468087411399505Aizawa, T., & Kosaka, H. (2008). Investigation of early soot formation process in a diesel spray flame via excitation—emission matrix using a multi-wavelength laser source. International Journal of Engine Research, 9(1), 79-97. doi:10.1243/14680874jer01407Bruneaux, G. (2008). Combustion structure of free and wall-impinging diesel jets by simultaneous laser-induced fluorescence of formaldehyde, poly-aromatic hydrocarbons, and hydroxides. International Journal of Engine Research, 9(3), 249-265. doi:10.1243/14680874jer0010

Evolution of Reproductive Morphology in Leaf Endophytes

The endophytic lifestyle has played an important role in the evolution of the morphology of reproductive structures (body) in one of the most problematic groups in fungal classification, the Leotiomycetes (Ascomycota). Mapping fungal morphologies to two groups in the Leiotiomycetes, the Rhytismatales and Hemiphacidiaceae reveals significant divergence in body size, shape and complexity. Mapping ecological roles to these taxa reveals that the groups include endophytic fungi living on leaves and saprobic fungi living on duff or dead wood. Finally, mapping of the morphologies to ecological roles reveals that leaf endophytes produce small, highly reduced fruiting bodies covered with fungal tissue or dead host tissue, while saprobic species produce large and intricate fruiting bodies. Intriguingly, resemblance between asexual conidiomata and sexual ascomata in some leotiomycetes implicates some common developmental pathways for sexual and asexual development in these fungi

Selection at a single locus leads to widespread expansion of toxoplasma gondii lineages that are virulent in mice

The determinants of virulence are rarely defined for eukaryotic parasites such as T. gondii, a widespread parasite of mammals that also infects humans, sometimes with serious consequences. Recent laboratory studies have established that variation in a single secreted protein, a serine/threonine kinase known as ROPO18, controls whether or not mice survive infection. Here, we establish the extent and nature of variation in ROP18among a collection of parasite strains from geographically diverse regions. Compared to other genes, ROP18 showed extremely high levels of diversification and changes in expression level, which correlated with severity of infection in mice. Comparison with an out-group demonstrated that changes in the upstream region that regulates expression of ROP18 led to an historical increase in the expression and exposed the protein to diversifying selective pressure. Surprisingly, only three atypically distinct protein variants exist despite marked genetic divergence elsewhere in the genome. These three forms of ROP18 are likely adaptations for different niches in nature, and they confer markedly different virulence to mice. The widespread distribution of a single mouse-virulent allele among geographically and genetically disparate parasites may have consequences for transmission and disease in other hosts, including humans