A prototype liquid Argon Time Projection Chamber for the study of UV laser multi-photonic ionization

This paper describes the design, realization and operation of a prototype liquid Argon Time Projection Chamber (LAr TPC) detector dedicated to the development of a novel online monitoring and calibration system exploiting UV laser beams. In particular, the system is intended to measure the lifetime of the primary ionization in LAr, in turn related to the LAr purity level. This technique could be exploited by present and next generation large mass LAr TPCs for which monitoring of the performance and calibration plays an important role. Results from the first measurements are presented together with some considerations and outlook.Comment: 26 pages, 27 figure

Preparative separation of DNA-ethidium bromide complexes by zonal density gradient centrifugation

Ethidium bromide-DNA complexes separated by rate-zonal sedimentation through a density gradient can be readily visualized and purified with little cross-contamination. The method is simple, rapid, and efficient.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/23311/1/0000249.pd

Increased Expression of the Auxiliary β(2)-subunit of Ventricular L-type Ca(2+) Channels Leads to Single-Channel Activity Characteristic of Heart Failure

BACKGROUND: Increased activity of single ventricular L-type Ca(2+)-channels (L-VDCC) is a hallmark in human heart failure. Recent findings suggest differential modulation by several auxiliary β-subunits as a possible explanation. METHODS AND RESULTS: By molecular and functional analyses of human and murine ventricles, we find that enhanced L-VDCC activity is accompanied by altered expression pattern of auxiliary L-VDCC β-subunit gene products. In HEK293-cells we show differential modulation of single L-VDCC activity by coexpression of several human cardiac β-subunits: Unlike β(1) or β(3) isoforms, β(2a) and β(2b) induce a high-activity channel behavior typical of failing myocytes. In accordance, β(2)-subunit mRNA and protein are up-regulated in failing human myocardium. In a model of heart failure we find that mice overexpressing the human cardiac Ca(V)1.2 also reveal increased single-channel activity and sarcolemmal β(2) expression when entering into the maladaptive stage of heart failure. Interestingly, these animals, when still young and non-failing (“Adaptive Phase”), reveal the opposite phenotype, viz : reduced single-channel activity accompanied by lowered β(2) expression. Additional evidence for the cause-effect relationship between β(2)-subunit expression and single L-VDCC activity is provided by newly engineered, double-transgenic mice bearing both constitutive Ca(V)1.2 and inducible β(2) cardiac overexpression. Here in non-failing hearts induction of β(2)-subunit overexpression mimicked the increase of single L-VDCC activity observed in murine and human chronic heart failure. CONCLUSIONS: Our study presents evidence of the pathobiochemical relevance of β(2)-subunits for the electrophysiological phenotype of cardiac L-VDCC and thus provides an explanation for the single L-VDCC gating observed in human and murine heart failure

Oxidant Sensing by Protein Kinases A and G Enables Integration of Cell Redox State with Phosphoregulation

The control of vascular smooth muscle contractility enables regulation of blood pressure, which is paramount in physiological adaptation to environmental challenges. Maintenance of stable blood pressure is crucial for health as deregulation (caused by high or low blood pressure) leads to disease progression. Vasotone is principally controlled by the cyclic nucleotide dependent protein kinases A and G, which regulate intracellular calcium and contractile protein calcium sensitivity. The classical pathways for activation of these two kinases are well established and involve the formation and activation by specific cyclic nucleotide second messengers. Recently we reported that both PKA and PKG can be regulated independently of their respective cyclic nucleotides via a mechanism whereby the kinases sense cellular oxidant production using redox active thiols. This novel redox regulation of these kinases is potentially of physiological importance, and may synergise with the classical regulatory mechanisms

Soil Quality - a critical review

Sampling and analysis or visual examination of soil to assess its status and use potential is widely practiced from plot to national scales. However, the choice of relevant soil attributes and interpretation of measurements are not straightforward, because of the complexity and site-specificity of soils, legacy effects of previous land use, and trade-offs between ecosystem services. Here we review soil quality and related concepts, in terms of definition, assessment approaches, and indicator selection and interpretation. We identify the most frequently used soil quality indicators under agricultural land use. We find that explicit evaluation of soil quality with respect to specific soil threats, soil functions and ecosystem services has rarely been implemented, and few approaches provide clear interpretation schemes of measured indicator values. This limits their adoption by land managers as well as policy. We also consider novel indicators that address currently neglected though important soil properties and processes, and we list the crucial steps in the development of a soil quality assessment procedure that is scientifically sound and supports management and policy decisions that account for the multi-functionality of soil. This requires the involvement of the pertinent actors, stakeholders and end-users to a much larger degree than practiced to date

Real-time visualization of heterotrimeric G protein Gq activation in living cells

Contains fulltext : 97296.pdf (publisher's version ) (Open Access)BACKGROUND: Gq is a heterotrimeric G protein that plays an important role in numerous physiological processes. To delineate the molecular mechanisms and kinetics of signalling through this protein, its activation should be measurable in single living cells. Recently, fluorescence resonance energy transfer (FRET) sensors have been developed for this purpose. RESULTS: In this paper, we describe the development of an improved FRET-based Gq activity sensor that consists of a yellow fluorescent protein (YFP)-tagged Ggamma2 subunit and a Galphaq subunit with an inserted monomeric Turquoise (mTurquoise), the best cyan fluorescent protein variant currently available. This sensor enabled us to determine, for the first time, the kon (2/s) of Gq activation. In addition, we found that the guanine nucleotide exchange factor p63RhoGEF has a profound effect on the number of Gq proteins that become active upon stimulation of endogenous histamine H1 receptors. The sensor was also used to measure ligand-independent activation of the histamine H1 receptor (H1R) upon addition of a hypotonic stimulus. CONCLUSIONS: Our observations reveal that the application of a truncated mTurquoise as donor and a YFP-tagged Ggamma2 as acceptor in FRET-based Gq activity sensors substantially improves their dynamic range. This optimization enables the real-time single cell quantification of Gq signalling dynamics, the influence of accessory proteins and allows future drug screening applications by virtue of its sensitivity

GoLoco motif proteins binding to Gαi1: insights from molecular simulations

Molecular dynamics simulations, computational alanine scanning and sequence analysis were used to investigate the structural properties of the Gαi1/GoLoco peptide complex. Using these methodologies, binding of the GoLoco motif peptide to the Gαi1 subunit was found to restrict the relative movement of the helical and catalytic domains in the Gαi1 subunit, which is in agreement with a proposed mechanism of GDP dissociation inhibition by GoLoco motif proteins. In addition, the results provide further insights into the role of the “Switch IV” region located within the helical domain of Gα, the conformation of which might be important for interactions with various Gα partners

A review of the impacts of degradation threats on soil properties in the UK

National governments are becoming increasingly aware of the importance of their soil resources and are shaping strategies accordingly. Implicit in any such strategy is that degradation threats and their potential effect on important soil properties and functions are defined and understood. In this paper, we aimed to review the principal degradation threats on important soil properties in the UK, seeking quantitative data where possible. Soil erosion results in the removal of important topsoil and, with it, nutrients, C and porosity. A decline in soil organic matter principally affects soil biological and microbiological properties, but also impacts on soil physical properties because of the link with soil structure. Soil contamination affects soil chemical properties, affecting nutrient availability and degrading microbial properties, whilst soil compaction degrades the soil pore network. Soil sealing removes the link between the soil and most of the ‘spheres’, significantly affecting hydrological and microbial functions, and soils on re-developed brownfield sites are typically degraded in most soil properties. Having synthesized the literature on the impact on soil properties, we discuss potential subsequent impacts on the important soil functions, including food and fibre production, storage of water and C, support for biodiversity, and protection of cultural and archaeological heritage. Looking forward, we suggest a twin approach of field-based monitoring supported by controlled laboratory experimentation to improve our mechanistic understanding of soils. This would enable us to better predict future impacts of degradation processes, including climate change, on soil properties and functions so that we may manage soil resources sustainably

Insensitivity of soil biological communities to phosphorus fertilization in intensively managed grassland systems

Efficient soil P cycling is essential for promoting optimal and sustainable grassland growth. The soil biological community is regarded as an important source of available P to the plant community. However, the effects of P fertilization on the soil biota are unclear. This study aimed to investigate the effects of P fertilization on plant and soil biological communities in two intensively managed grassland sites that had been receiving mineral-based P fertilization over a 14-year period. Both pastures had been frequently cut and harvested for plant material and had received only inorganic fertilizers from the establishment of the trial. Both sites were sampled four times from October 2009 to April 2011 and plant and a range of soil biological parameters were determined at each sampling period. The main findings of this study showed that soil chemical measures, such as labile inorganic P and total P concentrations, and plant yield and P contents responded as expected to P fertilization. However, all soil biological parameters either showed no response or inconsistent responses to P fertilization over the experimental period. This study indicates that intensive management regimes, for example intensive plant harvesting and fertilizer regimes, appear to override the relationship between plant and soil biological communities with respect to their response to P fertilization, and thus their productivity is apparently not predicated upon biotic activity.</p