Optimisation of recombinant production of active human cardiac SERCA2a ATPase

Methods for recombinant production of eukaryotic membrane proteins, yielding sufficient quantity and quality of protein for structural biology, remain a challenge. We describe here, expression and purification optimisation of the human SERCA2a cardiac isoform of Ca2+ translocating ATPase, using Saccharomyces cerevisiae as the heterologous expression system of choice. Two different expression vectors were utilised, allowing expression of C-terminal fusion proteins with a biotinylation domain or a GFP- His8 tag. Solubilised membrane fractions containing the protein of interest were purified onto Streptavidin-Sepharose, Ni-NTA or Talon resin, depending on the fusion tag present. Biotinylated protein was detected using specific antibody directed against SERCA2 and, advantageously, GFP-His8 fusion protein was easily traced during the purification steps using in-gel fluorescence. Importantly, talon resin affinity purification proved more specific than Ni-NTA resin for the GFP-His8 tagged protein, providing better separation of oligomers present, during size exclusion chromatography. The optimised method for expression and purification of human cardiac SERCA2a reported herein, yields purified protein (> 90%) that displays a calcium-dependent thapsigargin-sensitive activity and is suitable for further biophysical, structural and physiological studies. This work provides support for the use of Saccharomyces cerevisiae as a suitable expression system for recombinant production of multi-domain eukaryotic membrane proteins.

Dependence of sea ice yield-curve shape on ice thickness

In this note, the authors discuss the contribution that frictional sliding of ice floes (or floe aggregates) past each other and pressure ridging make to the plastic yield curve of sea ice. Using results from a previous study that explicitly modeled the amount of sliding and ridging that occurs for a given global strain rate, it is noted that the relative contribution of sliding and ridging to ice stress depends upon ice thickness. The implication is that the shape and size of the plastic yield curve is dependent upon ice thickness. The yield-curve shape dependence is in addition to plastic hardening/weakening that relates the size of the yield curve to ice thickness. In most sea ice dynamics models the yield-curve shape is taken to be independent of ice thickness. The authors show that the change of the yield curve due to a change in the ice thickness can be taken into account by a weighted sum of two thickness-independent rheologies describing ridging and sliding effects separately. It would be straightforward to implement the thickness-dependent yield-curve shape described here into sea ice models used for global or regional ice prediction.

Avoiding a bad apple: insect pollination enhances fruit quality and economic value

Insect pollination is important for food production globally and apples are one of the major fruit crops which are reliant on this ecosystem service. It is fundamentally important that the full range of benefits of insect pollination to crop production are understood, if the costs of interventions aiming to enhance pollination are to be compared against the costs of the interventions themselves. Most previous studies have simply assessed the benefits of pollination to crop yield and ignored quality benefits and how these translate through to economic values. In the present study we examine the influence of insect pollination services on farmgate output of two important UK apple varieties; Gala and Cox. Using field experiments, we quantify the influence of insect pollination on yield and importantly quality and whether either may be limited by sub-optimal insect pollination. Using an expanded bioeconomic model we value insect pollination to UK apple production and establish the potential for improvement through pollination service management. We show that insects are essential in the production of both varieties of apple in the UK and contribute a total of £36.7 million per annum, over £6 million more than the value calculated using more conventional dependence ratio methods. Insect pollination not only affects the quantity of production but can also have marked impacts on the quality of apples, influencing size, shape and effecting their classification for market. These effects are variety specific however. Due to the influence of pollination on both yield and quality in Gala, there is potential for insect pollination services to improve UK output by up to £5.7 million per annum. Our research shows that continued pollinator decline could have serious financial implications for the apple industry but there is considerable scope through management of wild pollinators or using managed pollinator augmentation, to improve the quality of production. Furthermore, we show that it is critically important to consider all production parameters including quality, varietal differences and management costs when valuing the pollination service of any crop so investment in pollinator management can be proportional to its contribution.

Pre-Columbian raised-field agriculture and land use in the Bolivian Amazon

We present an integrated palaeoecological and archaeobotanical study of pre-Columbian raised-field agriculture in the Llanos de Moxos, a vast seasonally inundated forest–savanna mosaic in the Bolivian Amazon. Phytoliths from excavated raised-field soil units, together with pollen and charcoal in sediment cores from two oxbow lakes, were analysed to provide a history of land use and agriculture at the El Cerro raised-field site. The construction of raised fields involved the removal of savanna trees, and gallery forest was cleared from the area by AD 310. Despite the low fertility of Llanos de Moxos soils, we determined that pre-Columbian raised-field agriculture sufficiently improved soil conditions for maize cultivation. Fire was used as a common management practice until AD 1300, at which point, the land-use strategy shifted towards less frequent burning of savannas and raised fields. Alongside a reduction in the use of fire, sweet potato cultivation and the exploitation of Inga fruits formed part of a mixed resource strategy from AD 1300 to 1450. The pre-Columbian impact on the landscape began to lessen around AD 1450, as shown by an increase in savanna trees and gallery forest. Although agriculture at the site began to decline prior to European arrival, the abandonment of raised fields was protracted, with evidence of sweet potato cultivation occurring as late as AD 1800.

Integrating control and electronics

Previously the author described how control engineering can be introduced using little mathematics in a first year course, the aim being to make the subject accessible across different degrees. One reaction to this was that it was a good idea, but there was not space to include it in the curriculum where, typically control engineering is not introduced until the second year. This paper describes how the author has used a review of the first year teaching to develop a module in which feedback, control and electronics are integrated coherently. This is beneficial as concepts in control and electronics mutually reinforce each other. This has been achieved during a reduction in the available time for teaching the material. This paper describes the strategy used to successfully develop the module, the integrated module and positive student reaction.

Science-based conservation and management in wetland archaeology: the example of Sutton Common, UK

This paper addresses the perception of different wetlands in and around the Humber estuary in the Bronze Age. Combining past and current research, it will be argued that the perception of intertidal wetlands was nearly diametrically opposed to the perception of riverine floodplains. This contrasting perception is reflected in the material culture of the Bronze Age, and may be explained through the particular manner in which landscapes changed following marine transgressions. This work was largely undertaken within the framework of the Humber Wetlands Survey, an integrated archaeological and palaeoenvironmental research programme funded by English Heritage since 1992

Polymer protected gold nanoparticles: synthesis, characterization and application in catalysis

This review discusses the stabilization of gold nanoparticles (AuNPs) by nonionic, anionic, cationic and amphoteric polymers. The protocols used for synthesis of AuNPs in aqueous and organic solvents are described. Size, shape and morphology of AuNPs are characterized by various physicochemical methods. Application aspects of polymer-protected AuNPs in catalysis are outlined.

Sources, sinks, and transatlantic transport of North African dust aerosol: a multimodel analysis and comparison with remote sensing data

This study evaluates model-simulated dust aerosols over North Africa and the North Atlantic from five global models that participated in the Aerosol Comparison between Observations and Models phase II model experiments. The model results are compared with satellite aerosol optical depth (AOD) data from Moderate Resolution Imaging Spectroradiometer (MODIS), Multiangle Imaging Spectroradiometer (MISR), and Sea-viewing Wide Field-of-view Sensor, dust optical depth (DOD) derived from MODIS and MISR, AOD and coarse-mode AOD (as a proxy of DOD) from ground-based Aerosol Robotic Network Sun photometer measurements, and dust vertical distributions/centroid height from Cloud Aerosol Lidar with Orthogonal Polarization and Atmospheric Infrared Sounder satellite AOD retrievals. We examine the following quantities of AOD and DOD: (1) the magnitudes over land and over ocean in our study domain, (2) the longitudinal gradient from the dust source region over North Africa to the western North Atlantic, (3) seasonal variations at different locations, and (4) the dust vertical profile shape and the AOD centroid height (altitude above or below which half of the AOD is located). The different satellite data show consistent features in most of these aspects; however, the models display large diversity in all of them, with significant differences among the models and between models and observations. By examining dust emission, removal, and mass extinction efficiency in the five models, we also find remarkable differences among the models that all contribute to the discrepancies of model-simulated dust amount and distribution. This study highlights the challenges in simulating the dust physical and optical processes, even in the best known dust environment, and stresses the need for observable quantities to constrain the model processes.

K-Surfer: a KNIME extension for the management and analysis of Human brain MRI FreeSurfer/FSL data

Human brain imaging techniques, such as Magnetic Resonance Imaging (MRI) or Diffusion Tensor Imaging (DTI), have been established as scientific and diagnostic tools and their adoption is growing in popularity. Statistical methods, machine learning and data mining algorithms have successfully been adopted to extract predictive and descriptive models from neuroimage data. However, the knowledge discovery process typically requires also the adoption of pre-processing, post-processing and visualisation techniques in complex data workflows. Currently, a main problem for the integrated preprocessing and mining of MRI data is the lack of comprehensive platforms able to avoid the manual invocation of preprocessing and mining tools, that yields to an error-prone and inefficient process. In this work we present K-Surfer, a novel plug-in of the Konstanz Information Miner (KNIME) workbench, that automatizes the preprocessing of brain images and leverages the mining capabilities of KNIME in an integrated way. K-Surfer supports the importing, filtering, merging and pre-processing of neuroimage data from FreeSurfer, a tool for human brain MRI feature extraction and interpretation. K-Surfer automatizes the steps for importing FreeSurfer data, reducing time costs, eliminating human errors and enabling the design of complex analytics workflow for neuroimage data by leveraging the rich functionalities available in the KNIME workbench.

Reverse correlation reveals how observers sample visual information when estimating three-dimensional shape

Human observers exhibit large systematic distance-dependent biases when estimating the three-dimensional (3D) shape of objects defined by binocular image disparities. This has led some to question the utility of disparity as a cue to 3D shape and whether accurate estimation of 3D shape is at all possible. Others have argued that accurate perception is possible, but only with large continuous perspective transformations of an object. Using a stimulus that is known to elicit large distance-dependent perceptual bias (random dot stereograms of elliptical cylinders) we show that contrary to these findings the simple adoption of a more naturalistic viewing angle completely eliminates this bias. Using behavioural psychophysics, coupled with a novel surface-based reverse correlation methodology, we show that it is binocular edge and contour information that allows for accurate and precise perception and that observers actively exploit and sample this information when it is available.

“Pussy cat, pussy cat where have you been..?” Evaluating the use of stable isotopes, radiogenic isotopes and trace elements for investigating the ecology of Panthera leo

1. Understanding the behaviour and ecology of large carnivores is becoming increasingly important as the list of endangered species grows, with felids such as Panthera leo in some locations heading dangerously close to extinction in the wild. In order to have more reliable and effective tools to understand animal behaviour, movement and diet, we need to develop novel, integrated approaches and effective techniques to capture a detailed profile of animal foraging and movement patterns. 2. Ecological studies have shown considerable interest in using stable isotope methods, both to investigate the nature of animal feeding habits, and to map their geographical location. However, recent work has suggested that stable isotope analyses of felid fur and bone is very complex and does not correlate directly with the isotopic composition of precipitation (and hence geographical location). 3. We present new data that suggest these previous findings may be atypical, and demonstrate that isotope analyses of Felidae are suitable for both evaluating dietary inputs and establishing geo-location as they have strong environmental referents to both food and water. These data provide new evidence of an important methodology that can be applied to the family Felidae for future research in ecology, conservation, wildlife forensics and archaeological science.