BASiCS: Bayesian Analysis of Single-Cell Sequencing Data

Single-cell mRNA sequencing can uncover novel cell-to-cell heterogeneity in gene expression levels in seemingly homogeneous populations of cells. However, these experiments are prone to high levels of unexplained technical noise, creating new challenges for identifying genes that show genuine heterogeneous expression within the population of cells under study. BASiCS (Bayesian Analysis of Single-Cell Sequencing data) is an integrated Bayesian hierarchical model where: (i) cell-specific normalisation constants are estimated as part of the model parameters, (ii) technical variability is quantified based on spike-in genes that are artificially introduced to each analysed cell's lysate and (iii) the total variability of the expression counts is decomposed into technical and biological components. BASiCS also provides an intuitive detection criterion for highly (or lowly) variable genes within the population of cells under study. This is formalised by means of tail posterior probabilities associated to high (or low) biological cell-to-cell variance contributions, quantities that can be easily interpreted by users. We demonstrate our method using gene expression measurements from mouse Embryonic Stem Cells. Cross-validation and meaningful enrichment of gene ontology categories within genes classified as highly (or lowly) variable supports the efficacy of our approach

Where is my sink? Reconstruction of landscape development in 1 southwestern Africa since the Late Jurassic

Quantifying the rates and timing of landscape denudation provides a means to constrain sediment flux through time to offshore sedimentary basins. The Late Mesozoic evolution of drainage basins in southern Africa is poorly constrained despite the presence of several onshore and offshore sedimentary basins. A novel approach has been developed to calculate the volume of material eroded since the Late Jurassic at different time steps by constructing structural cross-sections and extrapolating thicknesses of eroded material. Using different assumptions, the calculated volumes of material eroded from southwestern Africa range from 2.52x10⁶ km³ (11.3 km of vertical thickness removed) to 8.87 x10⁵ km³ (4.0 km of vertical thickness removed). For the southward draining systems alone, the calculated removal of 7.81 x10⁵ – 2.60 x10⁵ km³ of material is far greater than the volumes of sediment recorded in offshore sedimentary basins (268 500 km³). Reconstruction of the drainage systems using geomorphic indicators and clast provenance of the Uitenhage Group, as well as extrapolated surface exposure ages, indicate the southern draining systems were active from the Late Jurassic with coeval activity in axial and transverse drainage systems. The calculated volumes are tied to published apatite fission track (AFT) dates to constrain the changes in exhumation rate through time (using multiple scenarios), which indicate a significant amount of Early Cretaceous exhumation (up to 1.26 x10⁶ km³, equivalent to 5.70km of vertical thickness). For the first time, this has permitted long-term landscape evolution to be used to support the interpretation that some of the ‘missing’ sediment was deposited in sedimentary basins on the Falkland Plateau as it moved past southern Africa during the Early Cretaceous. This implies that in this instance, the sinks are separated from their source areas by ~6000 km

Shade Effects on Athletic Field Playing Characteristics of Overseeded and Non-Overseeded Bermudagrass

In many athletic stadiums around the world, reduced light levels from the stadium structure can significantly reduce turfgrass quality and playing characteristics. In most warm-weather stadiums, the primary surface is bermudagrass which is commonly overseeded with perennial ryegrass to provide a winter and spring playing surface. The objective of this study was to examine the effects of four shade levels on turfgrass quality and playing characteristics of overseeded and non-overseeded bermudagrass (cv. Riviera). Shade levels included a non-shaded control and shade treatments which blocked 30%, 60%, and 90% of ambient light. These shade levels resulted in average daily PAR loads of 45.1, 29.6, 16.7, and 4.7 mol PAR / m2 / day, respectively. Data collection included both quality and playability characteristics. The effects of shading became obvious as the study continued into early summer, with the 90% shade plots losing almost all cover and the 60% shade plots losing up to 40% of coverage through mid-summer. The 30% shade plots had a slight decline in turfgrass coverage, although, still retained close to 90% coverage. Based on these results, a minimum daily light load of approximately 30 mol PAR / m2 / day is needed to sustain Riviera bermudagrass at or near 100% coverage. In the overseeded plots, the only shade treatment that caused a significant reduction in turfgrass coverage was the 90% shade treatment (4.7 mol PAR / m2 / day). There was a significant decrease in the amount of bermudagrass present in the overseeded plots in mid-summer, suggesting that increased shading was causing a shift in population from the warm-season bermudagrass to the cool-season perennial ryegrass. In both overseeded and non-overseeded plots, rotational resistance (traction) decreased even with modest levels of shade , which is likely correlated to a decrease in turfgrass densit

Biochemical properties of Paracoccus denitrificans FnrP:Reactions with molecular oxygen and nitric oxide

In Paracoccus denitrificans, three CRP/FNR family regulatory proteins, NarR, NnrR and FnrP, control the switch between aerobic and anaerobic (denitrification) respiration. FnrP is a [4Fe-4S] cluster containing homologue of the archetypal O2 sensor FNR from E. coli and accordingly regulates genes encoding aerobic and anaerobic respiratory enzymes in response to O2, and also NO, availability. Here we show that FnrP undergoes O2-driven [4Fe-4S] to [2Fe-2S] cluster conversion that involves up to 2 O2 per cluster, with significant oxidation of released cluster sulfide to sulfane observed at higher O2 concentrations. The rate of the cluster reaction was found to be ~6-fold lower than that of E. coli FNR, suggesting that FnrP can remain transcriptionally active under microaerobic conditions. This is consistent with a role for FnrP in activating expression of the high O2 affinity cytochrome c oxidase under microaerobic conditions. Cluster conversion resulted in dissociation of the transcriptionally active FnrP dimer into monomers. Therefore, along with E. coli FNR, FnrP belongs to the subset of FNR proteins in which cluster type is correlated with association state. Interestingly, two key charged residues, Arg140 and Asp154, that have been shown to play key roles in the monomer-dimer equilibrium in E. coli FNR are not conserved in FnrP, indicating that different protomer interactions are important for this equilibrium. Finally, the FnrP [4Fe-4S] cluster is shown to undergo reaction with multiple NO molecules, resulting in iron nitrosyl species and dissociation into monomers

What is the impact of giant cell arteritis on patients' lives?: a UK qualitative study

Objectives: Clinical management of giant cell arteritis (GCA) involves balancing the risks and burdens arising from the disease with those arising from treatment, but there is little research on the nature of those burdens. We aimed to explore the impact of giant cell arteritis (GCA) and its treatment on patients’ lives. Methods: UK patients with GCA participated in semi-structured telephone interviews. Inductive thematic analysis was employed. Results: 24 participants were recruited (age: 65–92 years, time since diagnosis: 2 months to >6 years). The overarching themes from analysis were: ongoing symptoms of the disease and its treatment; and ‘life-changing’ impacts. The overall impact of GCA on patients’ lives arose from a changing combination of symptoms, side effects, adaptations to everyday life and impacts on sense of normality. Important factors contributing to loss of normality were glucocorticoid-related treatment burdens and fear about possible future loss of vision. Conclusions: The impact of GCA in patients’ everyday lives can be substantial, multifaceted and ongoing despite apparent control of disease activity. The findings of this study will help doctors better understand patient priorities, legitimise patients’ experiences of GCA and work with patients to set realistic treatment goals and plan adaptations to their everyday lives

A Genome-wide gene-expression analysis and database in transgenic mice during development of amyloid or tau pathology

We provide microarray data comparing genome-wide differential expression and pathology throughout life in four lines of "amyloid" transgenic mice (mutant human APP, PSEN1, or APP/PSEN1) and "TAU" transgenic mice (mutant human MAPT gene). Microarray data were validated by qPCR and by comparison to human studies, including genome-wide association study (GWAS) hits. Immune gene expression correlated tightly with plaques whereas synaptic genes correlated negatively with neurofibrillary tangles. Network analysis of immune gene modules revealed six hub genes in hippocampus of amyloid mice, four in common with cortex. The hippocampal network in TAU mice was similar except that Trem2 had hub status only in amyloid mice. The cortical network of TAU mice was entirely different with more hub genes and few in common with the other networks, suggesting reasons for specificity of cortical dysfunction in FTDP17. This Resource opens up many areas for investigation. All data are available and searchable at http://www.mouseac.org

Numb regulates cell–cell adhesion and polarity in response to tyrosine kinase signalling

Epithelial-mesenchymal transition (EMT), which can be caused by aberrant tyrosine kinase signalling, marks epithelial tumour progression and metastasis, yet the underlying molecular mechanism is not fully understood. Here, we report that Numb interacts with E-cadherin (E-cad) through its phosphotyrosine-binding domain (PTB) and thereby regulates the localization of E-cad to the lateral domain of epithelial cell–cell junction. Moreover, Numb engages the polarity complex Par3–aPKC–Par6 by binding to Par3 in polarized Madin-Darby canine kidney cells. Intriguingly, after Src activation or hepatocyte growth factor (HGF) treatment, Numb decouples from E-cad and Par3 and associates preferably with aPKC–Par6. Binding of Numb to aPKC is necessary for sequestering the latter in the cytosol during HGF-induced EMT. Knockdown of Numb by small hairpin RNA caused a basolateral-to-apicolateral translocation of E-cad and β-catenin accompanied by elevated actin polymerization, accumulation of Par3 and aPKC in the nucleus, an enhanced sensitivity to HGF-induced cell scattering, a decrease in cell–cell adhesion, and an increase in cell migration. Our work identifies Numb as an important regulator of epithelial polarity and cell–cell adhesion and a sensor of HGF signalling or Src activity during EMT