Novel approach for deriving genome wide SNP analysis data from archived blood spots.

RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are.BACKGROUND: The ability to transport and store DNA at room temperature in low volumes has the advantage of optimising cost, time and storage space. Blood spots on adapted filter papers are popular for this, with FTA (Flinders Technology Associates) Whatman™TM technology being one of the most recent. Plant material, plasmids, viral particles, bacteria and animal blood have been stored and transported successfully using this technology, however the method of porcine DNA extraction from FTA Whatman™TM cards is a relatively new approach, allowing nucleic acids to be ready for downstream applications such as PCR, whole genome amplification, sequencing and subsequent application to single nucleotide polymorphism microarrays has hitherto been under-explored. FINDINGS: DNA was extracted from FTA Whatman™TM cards (following adaptations of the manufacturer's instructions), whole genome amplified and subsequently analysed to validate the integrity of the DNA for downstream SNP analysis. DNA was successfully extracted from 288/288 samples and amplified by WGA. Allele dropout post WGA, was observed in less than 2% of samples and there was no clear evidence of amplification bias nor contamination. Acceptable call rates on porcine SNP chips were also achieved using DNA extracted and amplified in this way. CONCLUSIONS: DNA extracted from FTA Whatman cards is of a high enough quality and quantity following whole genomic amplification to perform meaningful SNP chip studies

Selective wheat germ agglutinin (WGA) uptake in the hippocampus from the locus coeruleus of dopamine-β-hydroxylase-WGA transgenic mice

We generated transgenic mice in which a trans-synaptic tracer, wheat germ agglutinin (WGA), was specifically expressed in the locus coeruleus (LC) neurons under the control of the dopamine-β-hydroxylase (DBH) gene promoter. WGA protein was produced in more than 95% of the tyrosine hydroxylase (TH)-positive LC neurons sampled. Transynaptic transfer of WGA was most evident in CA3 neurons of the hippocampus, but appeared absent in CA1 neurons. Faint but significant WGA immunoreactivity was observed surrounding the nuclei of dentate granule cells. Putative hilar mossy cells, identified by the presence of calretinin in the ventral hippocampus, appeared uniformly positive for transynaptically transferred WGA protein. GAD67-positive interneurons in the hilar and CA3 regions tended to be WGA-positive, although a subset of them did not show WGA co-localization. The same mixed WGA uptake profile was apparent when examining co-localization with parvalbumin. The selective uptake of WGA by dentate granule cells, mossy cells, and CA3 pyramidal neurons is consistent with evidence for a large proportion of conventional synapses adjacent to LC axonal varicosities in these regions. The lack of WGA uptake in the CA1 region and its relatively sparse innervation by DBH-positive fibers suggest that a majority of the TH-positive classical synapses revealed by electron microscopy in that region may be producing dopamine. The overall pattern of WGA uptake in these transgenic mice implies a selective role for the granule cell-mossy cell-CA3 network in processing novelty or the salient environmental contingency changes signaled by LC activity

Combining sediment source tracing techniques with traditional monitoring to assess the impact of improved land management on catchment sediment yields

Summary This paper aims to demonstrate the potential value of combining sediment source tracing techniques with traditional monitoring approaches, when documenting the impact of improved land management on catchment sediment yields. It reports the results of an investigation undertaken in a small (1.19 km 2 ) agricultural catchment in southern Brazil, which was monitored before and after the implementation of improved land management practices. Attention focussed on 50 storm events that occurred between May 2002 and March 2006 and which reflected the behaviour of the catchment during the pre-change, transition and post-change periods. Improved land management, involving minimum-till cultivation and the maintenance of good crop cover, was introduced in early 2003. The traditional monitoring provided a basis for evaluating the changes in storm runoff volume, storm hydrograph peak and storm-period sediment load and mean suspended sediment concentration. The results indicate that both storm runoff volumes and peak flows associated with a given amount of rainfall provided evidence of a significant decrease after the introduction of improved land management. Storm-period sediment loads showed a similar reduction, with a reduction by as much as 80% for low magnitude events and of ca. 40% for events of intermediate magnitude. However, there was no significant change in mean suspended sediment concentrations, indicating that the reductions in sediment load were primarily the result of the reduced storm runoff volume. Sediment source fingerprinting was used to explore the changes in the relative and absolute contributions to the storm sediment loads from the three key sources, Journal of Hydrology (2008) 348, 546-563 a v a i l a b l e a t w w w . s c i e n c e d i r e c t . c o m j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / j h y d r o l namely the surface of the fields under crops, the unpaved roads and the stream channels. A comparison of the load-weighted mean contributions for the pre-and post-treatment periods indicated that the contribution from the field surfaces and unpaved roads decreased from 63% and 36% to 54% and 24%, respectively, whereas the contribution from the stream channels increased from ca. 2% to 22%. By relating the absolute amounts of sediment mobilised from each individual source group to variables representing the runoff and precipitation associated with the events, it was possible to identify changes in the response of the individual sediment sources to the changes in land management that occurred within the catchment. Sediment mobilisation from the stream channel during individual events increased substantially over the whole range of flows after the introduction of improved land management in the study catchment, whereas the amounts of sediment mobilised from the surfaces of the fields and the unpaved roads showed a significant decrease during events of low and intermediate magnitude. The short monitoring period associated with the study, coupled with inter-annual variations in rainfall, necessarily limit the scope and rigour of the study reported, but it is seen to provide a useful demonstration of how the coupling of sediment source tracing with more traditional monitoring techniques can provide an improved understanding of the impact of improved management practices on the sediment response of a catchment, as well as important information to inform the design and implementation of effective sediment management and control measures.

Transport between edge states in multilayer integer quantum Hall systems: exact treatment of Coulomb interactions and disorder

A set of stacked two-dimensional electron systems in a perpendicular magnetic field exhibits a three-dimensional version of the quantum Hall effect if interlayer tunneling is not too strong. When such a sample is in a quantum Hall plateau, the edge states of each layer combine to form a chiral metal at the sample surface. We study the interplay of interactions and disorder in transport properties of the chiral metal, in the regime of weak interlayer tunneling. Our starting point is a system without interlayer tunneling, in which the only excitations are harmonic collective modes: surface magnetoplasmons. Using bosonization and working perturbatively in the interlayer tunneling amplitude, we express transport properties in terms of the spectrum for these collective modes, treating electron-electron interactions and impurity scattering exactly. We calculte the conductivity as a function of temperature, finding that it increases with increasing temperature as observed in recent experiments. We also calculate the autocorrelation function of mesoscopic conductance fluctuations induced by changes in a magnetic field component perpendicular to the sample surface, and its dependence on temperature. We show that conductance fluctuations are characterised by a dephasing length that varies inversely with temperature.Comment: 13 pages, 10 figures, minor changes made for publicatio

Phenotypic and genetic integration of personality and growth under competition in the sheepshead swordtail, Xiphophorus birchmanni

This is the author accepted manuscript. The final version is available from the publisher via the DOI in this record.Competition for resources including food, physical space, and potential mates is a fundamental ecological process shaping variation in individual phenotype and fitness. The evolution of competitive ability, in particular social dominance, depends on genetic (co)variation among traits causal (e.g., behaviour) or consequent (e.g. growth) to competitive outcomes. If dominance is heritable, it will generate both direct and indirect genetic effects (IGE) on resource dependent traits. The latter are expected to impose evolutionary constraint because winners necessarily gain resources at the expense of losers. We varied competition in a population of sheepshead swordtails, Xiphophorus birchmanni, to investigate effects on behaviour, size, growth, and survival. We then applied quantitative genetic analyses to determine (i) whether competition leads to phenotypic and/or genetic integration of behaviour with life history and (ii) the potential for IGE to constrain life history evolution. Size, growth and survival were reduced at high competition. Male dominance was repeatable and dominant individuals show higher growth and survival. Additive genetic contributions to phenotypic covariance were significant, with the G matrix largely recapitulating phenotypic relationships. Social dominance has a low but significant heritability and is strongly genetically correlated with size and growth. Assuming causal dependence of growth on dominance, hidden IGE will therefore reduce evolutionary potential.This work was supported by an EPSRC Studentship to KB, a BBSRC David Phillips Fellowship to AJW and a BBSRC grant (BB/L022656/1). CAW was funded by a NERC post-doctoral Research Fellowship (NE/I020245/1) and a University of Edinburgh Chancellor’s Fellowship. GGR was funded by the U.S. National Science Foundatio

Sensitivity of fluvial sediment source apportionment to mixing model assumptions: A Bayesian model comparison

Mixing models have become increasingly common tools for apportioning fluvial sediment load to various sediment sources across catchments using a wide variety of Bayesian and frequentist modeling approaches. In this study, we demonstrate how different model setups can impact upon resulting source apportionment estimates in a Bayesian framework via a one-factor-at-a-time (OFAT) sensitivity analysis. We formulate 13 versions of a mixing model, each with different error assumptions and model structural choices, and apply them to sediment geochemistry data from the River Blackwater, Norfolk, UK, to apportion suspended particulate matter (SPM) contributions from three sources (arable topsoils, road verges, and subsurface material) under base flow conditions between August 2012 and August 2013. Whilst all 13 models estimate subsurface sources to be the largest contributor of SPM (median ∼76%), comparison of apportionment estimates reveal varying degrees of sensitivity to changing priors, inclusion of covariance terms, incorporation of time-variant distributions, and methods of proportion characterization. We also demonstrate differences in apportionment results between a full and an empirical Bayesian setup, and between a Bayesian and a frequentist optimization approach. This OFAT sensitivity analysis reveals that mixing model structural choices and error assumptions can significantly impact upon sediment source apportionment results, with estimated median contributions in this study varying by up to 21% between model versions. Users of mixing models are therefore strongly advised to carefully consider and justify their choice of model structure prior to conducting sediment source apportionment investigations

Apportioning sources of organic matter in streambed sediments: An integrated molecular and compound-specific stable isotope approach

We present a novel application for quantitatively apportioning sources of organic matter in streambed sediments via a coupled molecular and compound-specific isotope analysis (CSIA) of long-chain leaf wax n-alkane biomarkers using a Bayesian mixing model. Leaf wax extracts of 13 plant species were collected from across two environments (aquatic and terrestrial) and four plant functional types (trees, herbaceous perennials, and C3 and C4 graminoids) from the agricultural River Wensum catchment, UK. Seven isotopic (δ13C27, δ13C29, δ13C31, δ13C27–31, δ2H27, δ2H29, and δ2H27–29) and two n-alkane ratio (average chain length (ACL), carbon preference index (CPI)) fingerprints were derived, which successfully differentiated 93% of individual plant specimens by plant functional type. The δ2H values were the strongest discriminators of plants originating from different functional groups, with trees (δ2H27–29 = − 208‰ to − 164‰) and C3 graminoids (δ2H27–29 = − 259‰ to − 221‰) providing the largest contrasts. The δ13C values provided strong discrimination between C3 (δ13C27–31 = − 37.5‰ to − 33.8‰) and C4 (δ13C27–31 = − 23.5‰ to − 23.1‰) plants, but neither δ13C nor δ2H values could uniquely differentiate aquatic and terrestrial species, emphasizing a stronger plant physiological/biochemical rather than environmental control over isotopic differences. ACL and CPI complemented isotopic discrimination, with significantly longer chain lengths recorded for trees and terrestrial plants compared with herbaceous perennials and aquatic species, respectively. Application of a comprehensive Bayesian mixing model for 18 streambed sediments collected between September 2013 and March 2014 revealed considerable temporal variability in the apportionment of organic matter sources. Median organic matter contributions ranged from 22% to 52% for trees, 29% to 50% for herbaceous perennials, 17% to 34% for C3 graminoids and 3% to 7% for C4 graminoids. The results presented here clearly demonstrate the effectiveness of an integrated molecular and stable isotope analysis for quantitatively apportioning, with uncertainty, plant-specific organic matter contributions to streambed sediments via a Bayesian mixing model approach

Attacks by a piercing-sucking insect (Myzus persicae Sultzer) or a chewing insect (Leptinotarsa decemlineata Say) on potato plants (Solanum tuberosum L.) induce differential changes in volatile compound release and oxylipin synthesis

Plant defensive strategies bring into play blends of compounds dependent on the type of attacker and coming from different synthesis pathways. Interest in the field is mainly focused on volatile organic compounds (VOCs) and jasmonic acid (JA). By contrast, little is known about the oxidized polyunsaturated fatty acids (PUFAs), such as PUFA-hydroperoxides, PUFA-hydroxides, or PUFA-ketones. PUFA-hydroperoxides and their derivatives might be involved in stress response and show antimicrobial activities. Hydroperoxides are also precursors of JA and some volatile compounds. In this paper, the differential biochemical response of a plant against insects with distinct feeding behaviours is characterized not only in terms of VOC signature and JA profile but also in terms of their precursors synthesized through the lipoxygenase (LOX)-pathway at the early stage of the plant response. For this purpose, two leading pests of potato with distinct feeding behaviours were used: the Colorado Potato Beetle (Leptinotarsa decemlineata Say), a chewing herbivore, and the Green Peach Aphid (Myzus persicae Sulzer), a piercing-sucking insect. The volatile signatures identified clearly differ in function with the feeding behaviour of the attacker and the aphid, which causes the smaller damages, triggers the emission of a higher number of volatiles. In addition, 9-LOX products, which are usually associated with defence against pathogens, were exclusively activated by aphid attack. Furthermore, a correlation between volatiles and JA accumulation and the evolution of their precursors was determined. Finally, the role of the insect itself on the plant response after insect infestation was highlighted