A functional proteomic method for the enrichment of peripheral membrane proteins reveals the collagen binding protein Hsp47 is exposed on the surface of activated human platelets

Platelets are small blood cells vital for hemostasis. Following vascular damage, platelets adhere to collagens and activate, forming a thrombus that plugs the wound and prevents blood loss. Stimulation of the platelet collagen receptor glycoprotein VI (GPVI) allows recruitment of proteins to receptor-proximal signaling complexes on the inner-leaflet of the plasma membrane. These proteins are often present at low concentrations; therefore, signaling-complex characterization using mass spectrometry is limited due to high sample complexity. We describe a method that facilitates detection of signaling proteins concentrated on membranes. Peripheral membrane proteins (reversibly associated with membranes) were eluted from human platelets with alkaline sodium carbonate. Liquid-phase isoelectric focusing and gel electrophoresis were used to identify proteins that changed in levels on membranes from GPVI-stimulated platelets. Immunoblot analysis verified protein recruitment to platelet membranes and subsequent protein phosphorylation was preserved. Hsp47, a collagen binding protein, was among the proteins identified and found to be exposed on the surface of GPVI-activated platelets. Inhibition of Hsp47 abolished platelet aggregation in response to collagen, while only partially reducing aggregation in response to other platelet agonists. We propose that Hsp47 may therefore play a role in hemostasis and thrombosis.

Methods and resources for climate impacts research: achieving synergy

The prediction of climate variability and change requires the use of a range of simulation models. Multiple climate model simulations are needed to sample the inherent uncertainties in seasonal to centennial prediction. Because climate models are computationally expensive, there is a tradeoff between complexity, spatial resolution, simulation length, and ensemble size. The methods used to assess climate impacts are examined in the context of this trade-off. An emphasis on complexity allows simulation of coupled mechanisms, such as the carbon cycle and feedbacks between agricultural land management and climate. In addition to improving skill, greater spatial resolution increases relevance to regional planning. Greater ensemble size improves the sampling of probabilities. Research from major international projects is used to show the importance of synergistic research efforts. The primary climate impact examined is crop yield, although many of the issues discussed are relevant to hydrology and health modeling. Methods used to bridge the scale gap between climate and crop models are reviewed. Recent advances include large-area crop modeling, quantification of uncertainty in crop yield, and fully integrated crop–climate modeling. The implications of trends in computer power, including supercomputers, are also discussed.

Reductions in insecticide use from adoption of Bt cotton in South Africa: impacts on economic performance and toxic load to the environment

The study reported presents the findings relating to commercial growing of genetically-modified Bt cotton in South Africa by a large sample of smallholder farmers over three seasons (1998/99, 1999/2000, 2000/01) following adoption. The analysis presents constructs and compares groupwise differences for key variables in Bt v. non-Bt technology and uses regressions to further analyse the production and profit impacts of Bt adoption. Analysis of the distribution of benefits between farmers due to the technology is also presented. In parallel with these socio-economic measures, the toxic loads being presented to the environment following the introduction of Bt cotton are monitored in terms of insecticide active ingredient (ai) and the Biocide Index. The latter adjusts ai to allow for differing persistence and toxicity of insecticides. Results show substantial and significant financial benefits to smallholder cotton growers of adopting Bt cotton over three seasons in terms of increased yields, lower insecticide spray costs and higher gross margins. This includes one particularly wet, poor growing season. In addition, those with the smaller holdings appeared to benefit proportionately more from the technology (in terms of higher gross margins) than those with larger holdings. Analysis using the Gini-coefficient suggests that the Bt technology has helped to reduce inequality amongst smallholder cotton growers in Makhathini compared to what may have been the position if they had grown conventional cotton. However, while Bt growers applied lower amounts of insecticide and had lower Biocide Indices (per ha) than growers of non-Bt cotton, some of this advantage was due to a reduction in non-bollworm insecticide. Indeed, the Biocide Index for all farmers in the population actually increased with the introduction of Bt cotton. The results indicate the complexity of such studies on the socio-economic and environmental impacts of GM varieties in the developing world.

A dendrochemical study of Pinus sylvestris from Siljansfors Experimental Forest, central Sweden

Inductively coupled plasma mass spectrometry (ICP-MS) was used to investigate changes in trace element concentration in two high resolution sequences of tree rings from central Sweden. Individual annual growth increments from 18002002 to 1930-2002 were sampled from two Scots pine (Pinus sylvestris) trees from the Siljansfors Experimental Forest. The aims of the study were: to test the viability of conventional solution induction ICP-MS as a technique for investigating the multi-elemental chemistry of long tree ring sequences at annual resolution, and, to test this specifically with a view to detecting changes in elemental concentrations of Swedish tree rings contemporary with the major (and relatively proximal) Icelandic eruption of Askja (1875). It was found that despite a time consuming sample preparation process, it was possible to use conventional ICP-MS for multi-elemental analysis of a long sequence of tree rings at annual resolution. Although promising data were produced, no truly conclusive concentration anomaly could be detected in the sequence to indicate the impact of the Askja eruption on environmental chemistry. Overall findings underlined the complexity of the tree/environment interaction and the cautious approach to data interpretation essential for any dendrochemical study. (c) 2006 Elsevier Ltd. All rights reserved.

GC/multiple collector-ICPMS method for chlorine stable isotope analysis of chlorinated aliphatic hydrocarbons

Stable isotopic characterization of chlorine in chlorinated aliphatic pollution is potentially very valuable for risk assessment and monitoring remediation or natural attenuation. The approach has been underused because of the complexity of analysis and the time it takes. We have developed a new method that eliminates sample preparation. Gas chromatography produces individually eluted sample peaks for analysis. The He carrier gas is mixed with Ar and introduced directly into the torch of a multicollector ICPMS. The MC-ICPMS is run at a high mass resolution of >= 10 000 to eliminate interference of mass 37 ArH with Cl. The standardization approach is similar to that for continuous flow stable isotope analysis in which sample and reference materials are measured successively. We have measured PCE relative to a laboratory TCE standard mixed with the sample. Solvent samples of 200 nmol to 1.3 mu mol ( 24- 165 mu g of Cl) were measured. The PCE gave the same value relative to the TCE as measured by the conventional method with a precision of 0.12% ( 2 x standard error) but poorer precision for the smaller samples.

The influence of project complexity on estimating accuracy

With the rapid development in technology over recent years, construction, in common with many areas of industry, has become increasingly complex. It would, therefore, seem to be important to develop and extend the understanding of complexity so that industry in general and in this case the construction industry can work with greater accuracy and efficiency to provide clients with a better service. This paper aims to generate a definition of complexity and a method for its measurement in order to assess its influence upon the accuracy of the quantity surveying profession in UK new build office construction. Quantitative data came from an analysis of twenty projects of varying size and value and qualitative data came from interviews with professional quantity surveyors. The findings highlight the difficulty in defining and measuring project complexity. The correlation between accuracy and complexity was not straightforward, being subjected to many extraneous variables, particularly the impact of project size. Further research is required to develop a better measure of complexity. This is in order to improve the response of quantity surveyors, so that an appropriate level of effort can be applied to individual projects, permitting greater accuracy and enabling better resource planning within the profession.

The use of bootstrapped Malmquist indices to reassess productivity change findings: an application to a sample of Polish farms

This article assesses the extent to which sampling variation affects findings about Malmquist productivity change derived using data envelopment analysis (DEA), in the first stage by calculating productivity indices and in the second stage by investigating the farm-specific change in productivity. Confidence intervals for Malmquist indices are constructed using Simar and Wilson's (1999) bootstrapping procedure. The main contribution of this article is to account in the second stage for the information in the second stage provided by the first-stage bootstrap. The DEA SEs of the Malmquist indices given by bootstrapping are employed in an innovative heteroscedastic panel regression, using a maximum likelihood procedure. The application is to a sample of 250 Polish farms over the period 1996 to 2000. The confidence intervals' results suggest that the second half of 1990s for Polish farms was characterized not so much by productivity regress but rather by stagnation. As for the determinants of farm productivity change, we find that the integration of the DEA SEs in the second-stage regression is significant in explaining a proportion of the variance in the error term. Although our heteroscedastic regression results differ with those from the standard OLS, in terms of significance and sign, they are consistent with theory and previous research.

Reductions in insecticide use from adoption of Bt cotton in South Africa: impacts on economic performance and toxic load to the environment

The study reported presents the findings relating to commercial growing of genetically-modified Bt cotton in South Africa by a large sample of smallholder farmers over three seasons (1998/99, 1999/2000, 2000/01) following adoption. The analysis presents constructs and compares groupwise differences for key variables in Bt v. non-Bt technology and uses regressions to further analyse the production and profit impacts of Bt adoption. Analysis of the distribution of benefits between farmers due to the technology is also presented. In parallel with these socio-economic measures, the toxic loads being presented to the environment following the introduction of Bt cotton are monitored in terms of insecticide active ingredient (ai) and the Biocide Index. The latter adjusts ai to allow for differing persistence and toxicity of insecticides. Results show substantial and significant financial benefits to smallholder cotton growers of adopting Bt cotton over three seasons in terms of increased yields, lower insecticide spray costs and higher gross margins. This includes one particularly wet, poor growing season. In addition, those with the smaller holdings appeared to benefit proportionately more from the technology (in terms of higher gross margins) than those with larger holdings. Analysis using the Gini-coefficient suggests that the Bt technology has helped to reduce inequality amongst smallholder cotton growers in Makhathini compared to what may have been the position if they had grown conventional cotton. However, while Bt growers applied lower amounts of insecticide and had lower Biocide Indices (per ha) than growers of non-Bt cotton, some of this advantage was due to a reduction in non-bollworm insecticide. Indeed, the Biocide Index for all farmers in the population actually increased with the introduction of Bt cotton. The results indicate the complexity of such studies on the socio-economic and environmental impacts of GM varieties in the developing world.

An exploration of the drivers to bio-security collective action among a sample of UK cattle and sheep farmers

At present, collective action regarding bio-security among UK cattle and sheep farmers is rare. Despite the occurrence of catastrophic livestock diseases such as bovine spongiform encephalopathy (BSE) and foot and mouth disease (FMD), within recent decades, there are few national or local farmer-led animal health schemes. To explore the reasons for this apparent lack of interest, we utilised a socio-psychological approach to disaggregate the cognitive, emotive and contextual factors driving bio-security behaviour among cattle and sheep farmers in the United Kingdom (UK). In total, we interviewed 121 farmers in South-West England and Wales. The main analytical tools included a content, cluster and logistic regression analysis. The results of the content analysis illustrated apparent 'dissonance' between bio-security attitudes and behaviour.(1) Despite the heavy toll animal disease has taken on the agricultural economy, most study participants were dismissive of the many measures associated with bio-security. Justification for this lack of interest was largely framed in relation to the collective attribution or blame for the disease threats themselves. Indeed, epidemic diseases were largely related to external actors and agents. Reasons for outbreaks included inadequate border control, in tandem with ineffective policies and regulations. Conversely, endemic livestock disease was viewed as a problem for 'bad' farmers and not an issue for those individuals who managed their stock well. As such, there was little utility in forming groups to address what was largely perceived as an individual problem. Further, we found that attitudes toward bio-security did not appear to be influenced by any particular source of information per se. While strong negative attitudes were found toward specific sources of bio-security information, e.g. government leaflets, these appear to simply reflect widely held beliefs. In relation to actual bio-security behaviours, the logistic regression analysis revealed no significant difference between in-scheme and out of scheme farmers. We concluded that in order to support collective action with regard to bio-security, messages need to be reframed and delivered from a neutral source. Efforts to support group formation must also recognise and address the issues relating to perceptions of social connectedness among the communities involved. (c) 2008 Elsevier B.V. All rights reserved.