Error, reproducibility and sensitivity: a pipeline for data processing of Agilent oligonucleotide expression arrays

Background: Expression microarrays are increasingly used to obtain large scale transcriptomic information on a wide range of biological samples. Nevertheless, there is still much debate on the best ways to process data, to design experiments and analyse the output. Furthermore, many of the more sophisticated mathematical approaches to data analysis in the literature remain inaccessible to much of the biological research community. In this study we examine ways of extracting and analysing a large data set obtained using the Agilent long oligonucleotide transcriptomics platform, applied to a set of human macrophage and dendritic cell samples. Results: We describe and validate a series of data extraction, transformation and normalisation steps which are implemented via a new R function. Analysis of replicate normalised reference data demonstrate that intrarray variability is small (only around 2 of the mean log signal), while interarray variability from replicate array measurements has a standard deviation (SD) of around 0.5 log(2) units (6 of mean). The common practise of working with ratios of Cy5/Cy3 signal offers little further improvement in terms of reducing error. Comparison to expression data obtained using Arabidopsis samples demonstrates that the large number of genes in each sample showing a low level of transcription reflect the real complexity of the cellular transcriptome. Multidimensional scaling is used to show that the processed data identifies an underlying structure which reflect some of the key biological variables which define the data set. This structure is robust, allowing reliable comparison of samples collected over a number of years and collected by a variety of operators. Conclusions: This study outlines a robust and easily implemented pipeline for extracting, transforming normalising and visualising transcriptomic array data from Agilent expression platform. The analysis is used to obtain quantitative estimates of the SD arising from experimental (non biological) intra- and interarray variability, and for a lower threshold for determining whether an individual gene is expressed. The study provides a reliable basis for further more extensive studies of the systems biology of eukaryotic cells.

A universal agro-hydrological model for water and nitrogen cycles in the soil-crop system SMCR_N: critical update and further validation

Agro-hydrological models have widely been used for optimizing resources use and minimizing environmental consequences in agriculture. SMCRN is a recently developed sophisticated model which simulates crop response to nitrogen fertilizer for a wide range of crops, and the associated leaching of nitrate from arable soils. In this paper, we describe the improvements of this model by replacing the existing approximate hydrological cascade algorithm with a new simple and explicit algorithm for the basic soil water flow equation, which not only enhanced the model performance in hydrological simulation, but also was essential to extend the model application to the situations where the capillary flow is important. As a result, the updated SMCRN model could be used for more accurate study of water dynamics in the soil-crop system. The success of the model update was demonstrated by the simulated results that the updated model consistently out-performed the original model in drainage simulations and in predicting time course soil water content in different layers in the soil-wheat system. Tests of the updated SMCRN model against data from 4 field crop experiments showed that crop nitrogen offtakes and soil mineral nitrogen in the top 90 cm were in a good agreement with the measured values, indicating that the model could make more reliable predictions of nitrogen fate in the crop-soil system, and thus provides a useful platform to assess the impacts of nitrogen fertilizer on crop yield and nitrogen leaching from different production systems. (C) 2010 Elsevier B.V. All rights reserved.

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.

Further investigations of the quartz optically stimulated luminescence components using linear modulation

The optically stimulated luminescence (OSL) from quartz is known to be the sum of several components with different rates of charge loss, originating from different trap types. The OSL components are clearly distinguished using the linear modulation (LM OSL) technique. A variety of pre-treatment and measurement conditions have been used on sedimentary samples in conjunction with linearly modulated optical stimulation to study in detail the behaviour of the OSL components of quartz. Single aliquots of different quartz samples have been found to contain typically five or six common LM OSL components when stimulated at View the MathML source. The components have been parameterised in terms of thermal stability (i.e. E and s), photoionisation cross-section energy dependence and dose response. The results of studies concerning applications of component-resolved LM OSL measurements on quartz are also presented. These include the detection of partial bleaching in young samples, use of ‘stepped wavelength’ stimulation to observe OSL from single components and attempts to extend the age range of quartz OSL dating.

Further evidence for variable synchrotron emission in XTE J1118+480 in outburst

We present simultaneous multicolor infrared and optical photometry of the black hole X-ray transient XTE J1118+480 during its short 2005 January outburst, supported by simultaneous X-ray observations. The variability is dominated by short timescales, ~10 s, although a weak superhump also appears to be present in the optical. The optical rapid variations, at least, are well correlated with those in X-rays. Infrared JHKs photometry, as in the previous outburst, exhibits especially large-amplitude variability. The spectral energy distribution (SED) of the variable infrared component can be fitted with a power law of slope α=-0.78+/-0.07, where F_ν~ν^α. There is no compelling evidence for evolution in the slope over five nights, during which time the source brightness decayed along almost the same track as seen in variations within the nights. We conclude that both short-term variability and longer timescale fading are dominated by a single component of constant spectral shape. We cannot fit the SED of the IR variability with a credible thermal component, either optically thick or thin. This IR SED is, however, approximately consistent with optically thin synchrotron emission from a jet. These observations therefore provide indirect evidence to support jet-dominated models for XTE J1118+480 and also provide a direct measurement of the slope of the optically thin emission, which is impossible, based on the average spectral energy distribution alone.

Further clarifying the competition–performance relation: reply to D. W. Johnson et al. (2012)

In their commentary, D. W. Johnson, Johnson, and Roseth (2012) provided some laudatory statements about our article, but they also expressed a number of concerns. The concerns focus on the following issues: types and definitions of competition, our choice of control group, the nature of performance-approach and performance-avoidance goals, the comprehensiveness of the opposing processes model, and performance-approach goals and constructive competition. We respond to each of these issues in turn and conclude with a statement regarding working to build an integrative model of the competition–performance relation (and beyond).

The impact of climate change on the treatability of dissolved organic matter (DOM) in upland water supplies: A UK perspective

Climate change in the UK is expected to cause increases in temperatures, altered precipitation patterns and more frequent and extreme weather events. In this review we discuss climate effects on dissolved organic matter (DOM), how altered DOM and water physico-chemical properties will affect treatment processes and assess the utility of techniques used to remove DOM and monitor water quality. A critical analysis of the literature has been undertaken with a focus on catchment drivers of DOM character, removal of DOM via coagulation and the formation of disinfectant by-products (DBPs). We suggest that: (1) upland catchments recovering from acidification will continue to produce more DOM with a greater hydrophobic fraction as solubility controls decrease; (2) greater seasonality in DOM export is likely in future due to altered precipitation patterns; (3) changes in species diversity and water properties could encourage algal blooms; and (4) that land management and vegetative changes may have significant effects on DOM export and treatability but require further research. Increases in DBPs may occur where catchments have high influence from peatlands or where algal blooms become an issue. To increase resilience to variable DOM quantity and character we suggest that one or more of the following steps are undertaken at the treatment works: a) ‘enhanced coagulation’ optimised for DOM removal; b) switching from aluminium to ferric coagulants and/or incorporating coagulant aids; c) use of magnetic ion-exchange (MIEX) pre-coagulation; and d) activated carbon filtration post-coagulation. Fluorescence and UV absorbance techniques are highlighted as potential methods for low-cost, rapid on-line process optimisation to improve DOM removal and minimise DBPs.

A systematic approach to identify the sources of tropical SST errors in coupled models using the adjustment of initialised experiments

Understanding the sources of systematic errors in climate models is challenging because of coupled feedbacks and errors compensation. The developing seamless approach proposes that the identification and the correction of short term climate model errors have the potential to improve the modeled climate on longer time scales. In previous studies, initialised atmospheric simulations of a few days have been used to compare fast physics processes (convection, cloud processes) among models. The present study explores how initialised seasonal to decadal hindcasts (re-forecasts) relate transient week-to-month errors of the ocean and atmospheric components to the coupled model long-term pervasive SST errors. A protocol is designed to attribute the SST biases to the source processes. It includes five steps: (1) identify and describe biases in a coupled stabilized simulation, (2) determine the time scale of the advent of the bias and its propagation, (3) find the geographical origin of the bias, (4) evaluate the degree of coupling in the development of the bias, (5) find the field responsible for the bias. This strategy has been implemented with a set of experiments based on the initial adjustment of initialised simulations and exploring various degrees of coupling. In particular, hindcasts give the time scale of biases advent, regionally restored experiments show the geographical origin and ocean-only simulations isolate the field responsible for the bias and evaluate the degree of coupling in the bias development. This strategy is applied to four prominent SST biases of the IPSLCM5A-LR coupled model in the tropical Pacific, that are largely shared by other coupled models, including the Southeast Pacific warm bias and the equatorial cold tongue bias. Using the proposed protocol, we demonstrate that the East Pacific warm bias appears in a few months and is caused by a lack of upwelling due to too weak meridional coastal winds off Peru. The cold equatorial bias, which surprisingly takes 30 years to develop, is the result of an equatorward advection of midlatitude cold SST errors. Despite large development efforts, the current generation of coupled models shows only little improvement. The strategy proposed in this study is a further step to move from the current random ad hoc approach, to a bias-targeted, priority setting, systematic model development approach.

The surface temperatures of Earth: steps towards integrated understanding of variability and change

Surface temperature is a key aspect of weather and climate, but the term may refer to different quantities that play interconnected roles and are observed by different means. In a community-based activity in June 2012, the EarthTemp Network brought together 55 researchers from five continents to improve the interaction between scientific communities who focus on surface temperature in particular domains, to exploit the strengths of different observing systems and to better meet the needs of different communities. The workshop identified key needs for progress towards meeting scientific and societal requirements for surface temperature understanding and information, which are presented in this community paper. A "whole-Earth" perspective is required with more integrated, collaborative approaches to observing and understanding Earth's various surface temperatures. It is necessary to build understanding of the relationships between different surface temperatures, where presently inadequate, and undertake large-scale systematic intercomparisons. Datasets need to be easier to obtain and exploit for a wide constituency of users, with the differences and complementarities communicated in readily understood terms, and realistic and consistent uncertainty information provided. Steps were also recommended to curate and make available data that are presently inaccessible, develop new observing systems and build capacities to accelerate progress in the accuracy and usability of surface temperature datasets.

Numerical analyses of Runge–Kutta implicit–explicit schemes for horizontally explicit, vertically implicit solutions of atmospheric models

With the prospect of exascale computing, computational methods requiring only local data become especially attractive. Consequently, the typical domain decomposition of atmospheric models means horizontally-explicit vertically-implicit (HEVI) time-stepping schemes warrant further attention. In this analysis, Runge-Kutta implicit-explicit schemes from the literature are analysed for their stability and accuracy using a von Neumann stability analysis of two linear systems. Attention is paid to the numerical phase to indicate the behaviour of phase and group velocities. Where the analysis is tractable, analytically derived expressions are considered. For more complicated cases, amplification factors have been numerically generated and the associated amplitudes and phase diagnosed. Analysis of a system describing acoustic waves has necessitated attributing the three resultant eigenvalues to the three physical modes of the system. To do so, a series of algorithms has been devised to track the eigenvalues across the frequency space. The result enables analysis of whether the schemes exactly preserve the non-divergent mode; and whether there is evidence of spurious reversal in the direction of group velocities or asymmetry in the damping for the pair of acoustic modes. Frequency ranges that span next-generation high-resolution weather models to coarse-resolution climate models are considered; and a comparison is made of errors accumulated from multiple stability-constrained shorter time-steps from the HEVI scheme with a single integration from a fully implicit scheme over the same time interval. Two schemes, “Trap2(2,3,2)” and “UJ3(1,3,2)”, both already used in atmospheric models, are identified as offering consistently good stability and representation of phase across all the analyses. Furthermore, according to a simple measure of computational cost, “Trap2(2,3,2)” is the least expensive.

Review of recent research (1998-2012) in German for Academic Purposes (GAP) in comparison with English for Academic Purposes (EAP): cross-influences, synergies and implications for further research

This state-of-the-art review reports on the major studies conducted in the field of Deutsch als Wissenschaftssprache (academic German) since the late 1990s. To begin with, the current position of German as a language of academic communication nationally and internationally will be discussed, focusing especially on the challenges posed by the status of English as a lingua franca. Subsequently, the major research undertaken since the late 1990s will be reviewed and its contribution to the development of teaching materials evaluated. Since studies on academic German have been influenced, to some extent, by research in English for Academic Purposes (EAP), this paper also attempts to dovetail developments in EAP in order to highlight commonalties and differences. The final sections will discuss some potential synergies and implications for further research in both fields.

Scoping customer relationship management strategy in HEI - Understanding steps towards alignment of customer and management needs

Higher Education Institutions (HEI) are complex organisations, offering a wide range of services, which involve a multiplicity of customers, stakeholders and service providers; both in terms of type and number. Satisfying a diverse set of customer groups is complex, and requires development of strategic Customer Relationship Management (CRM). This paper contributes to the HEI area, by proposing an approach that scopes CRM strategy, allowing us a better understanding CRM implementation in Higher Education Institutions; maximising alignment of customer and management desires, expectation and needs.