PCR for the detection of pathogens in neonatal early onset sepsis.

BACKGROUND: A large proportion of neonates are treated for presumed bacterial sepsis with broad spectrum antibiotics even though their blood cultures subsequently show no growth. This study aimed to investigate PCR-based methods to identify pathogens not detected by conventional culture. METHODS: Whole blood samples of 208 neonates with suspected early onset sepsis were tested using a panel of multiplexed bacterial PCRs targeting Streptococcus pneumoniae, Streptococcus agalactiae (GBS), Staphylococcus aureus, Streptococcus pyogenes (GAS), Enterobacteriaceae, Enterococcus faecalis, Enterococcus faecium, Ureaplasma parvum, Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium, a 16S rRNA gene broad-range PCR and a multiplexed PCR for Candida spp. RESULTS: Two-hundred and eight samples were processed. In five of those samples, organisms were detected by conventional culture; all of those were also identified by PCR. PCR detected bacteria in 91 (45%) of the 203 samples that did not show bacterial growth in culture. S. aureus, Enterobacteriaceae and S. pneumoniae were the most frequently detected pathogens. A higher bacterial load detected by PCR was correlated positively with the number of clinical signs at presentation. CONCLUSION: Real-time PCR has the potential to be a valuable additional tool for the diagnosis of neonatal sepsis

Multi-Stage Magma Evolution in Intra-Plate Volcanoes: Insights From Combined in situ Li and Mg–Fe Chemical and Isotopic Diffusion Profiles in Olivine

Understanding the timescales of magma evolution and ascent is essential for interpreting geophysical monitoring signals from active volcanoes. In this study, we explore the potential of diffusion-driven Li concentration and isotope zoning profiles recorded by magmatic olivine crystals to unravel time scales of magma evolution processes. Lithium is a fast-diffusing element and may provide the opportunity to investigate changes in magma composition during magma ascent, shortly before eruption. Lithium chemical and isotopic profiles were determined in olivines from two localities in the Massif Central volcanic region (France) that have previously been investigated for their Fe–Mg isotope systematics. The combined investigation of isotopic and chemical profiles makes it possible to distinguish between crystal growth and diffusion events. Extremely low δ7Li-values down to −30.7‰ (relative to the commonly used Li isotope standard IRMM-16) in the crystal core regions and elevated values at crystal rims (δ7Li ∼8 to 10‰), along with increasing concentrations from cores (∼3 to 1 μg/g) toward rims (12 to 6 μg/g) were found. The shape and orientation of both the chemical and isotopic profiles indicate that they were dominantly generated by Li diffusion into and within the olivine grains during magmatic differentiation. While Mg–Fe isotope and major element profiles have been modeled by a single diffusion event (Oeser et al., 2015), concentration and isotope profiles of Li indicate that a second diffusion event took place, that was not recorded by the Mg–Fe exchange diffusion couple. The first diffusion event was interpreted as reflecting the residence of the olivine crystals in a magma chamber. As diffusion coefficients for Fe–Mg exchange diffusion are very well determined, the time scales of this event are likely best quantified by Mg–Fe isotopic exchange diffusion modeling (Oeser et al., 2015). This event probably also generated the low δ7Li observed in olivine cores. Comparing the length of the Mg–Fe and Li profiles could thus be used to determine the less well-known diffusion coefficients of Li in the studied olivine crystals. The findings of this study indicate that Li diffusion at low Li concentration levels, as typically observed in natural olivine, may be not as fast as previously thought. The second diffusion event might represent a short-lived event, such as degassing, related to the ascent of the magma and/or magma cooling after emplacement of the lava. Such a process would only affect Li, which, in contrast to the refractory elements Fe and Mg, is volatile during degassing. The findings of this study show that, according to their different diffusion rates and physiochemical properties, the combined use of spatially resolved Li and Mg–Fe chemical and isotopic diffusion profiles, is a powerful tool to model even multi-stage evolution processes in magmatic systems. © Copyright © 2020 Steinmann, Oeser, Horn and Weyer

Exploring the Effect of G6PC2 Single Nucleotide Polymorphisms on Enzyme Activity and Human Health

G6PC2 encodes a glucose-6-phosphatase catalytic subunit that is highly expressed in pancreatic islet beta cells. Genome wide association studies (GWAS) have shown that single nucleotide polymorphisms (SNPs) in the G6PC2 gene are associated with variations in fasting blood glucose (FBG), a parameter linked with risk for type 2 diabetes (T2D). Studies in mice have complemented these GWAS data by showing that deletion of G6pc2 abolishes islet glucose-6-phosphatase activity and lowers FBG. We hypothesize that G6pc2 forms a substrate cycle with glucokinase that determines the sensitivity of glucose-stimulated insulin secretion (GSIS) to glucose. In support of this hypothesis we have previously shown that deletion of G6pc2 enhances GSIS at sub-maximal glucose concentrations and abolishes glucose cycling in isolated islets. More recently we have demonstrated that deletion of G6pc2 enhances glycolysis in isolated mouse islets, and that high rates of glucose cycling are also detected in human islets. Our broad hypothesis is that the results of these studies will strongly suggest that G6PC2 inhibition should be considered as a novel therapeutic strategy for lowering FBG and thereby preventing T2D. To extend these observations we have developed a novel intact cell assay for G6PC2 activity. This assay relies on the observation that CREB and ChREBP bound to the rat G6PC1 promoter are highly glucose responsive in the rat islet-derived 832/13 cell line and the fact that endogenous G6PC2 is absent. In the presence of catalytically-dead G6PC2, glucose stimulates G6PC1-luciferase fusion gene expression. However, this induction is blunted in the presence of wild type G6PC2. We are using this assay to determine the effect of non-synonymous G6PC2 SNPs on G6PC2 activity and then examining the association between SNPs that markedly affect G6PC2 activity with their effects on human health as assessed using Vanderbilt’s BioVU biobank. These data will reveal whether SNPs in G6PC2 are associated with only altered FBG or whether G6PC2 affects other aspects of human health

Surveillance of congenital Zika syndrome in England and Wales: methods and results of laboratory, obstetric and paediatric surveillance.

The spread of the Zika virus (ZIKV) in the Americas led to large outbreaks across the region and most of the Southern hemisphere. Of greatest concern were complications following acute infection during pregnancy. At the beginning of the outbreak, the risk to unborn babies and their clinical presentation was unclear. This report describes the methods and results of the UK surveillance response to assess the risk of ZIKV to children born to returning travellers. Established surveillance systems operating within the UK - the paediatric and obstetric surveillance units for rare diseases, and national laboratory monitoring - enabled rapid assessment of this emerging public health threat. A combined total of 11 women experiencing adverse pregnancy outcomes after possible ZIKV exposure were reported by the three surveillance systems; five miscarriages, two intrauterine deaths and four children with clinical presentations potentially associated with ZIKV infection. Sixteen women were diagnosed with ZIKV during pregnancy in the UK. Amongst the offspring of these women, there was unequivocal laboratory evidence of infection in only one child. In the UK, the number and risk of congenital ZIKV infection for travellers returning from ZIKV-affected countries is very small

Genome wide profiling of human embryonic stem cells (hESCs), their derivatives and embryonal carcinoma cells to develop base profiles of U.S. Federal government approved hESC lines

BACKGROUND: In order to compare the gene expression profiles of human embryonic stem cell (hESC) lines and their differentiated progeny and to monitor feeder contaminations, we have examined gene expression in seven hESC lines and human fibroblast feeder cells using Illumina(® )bead arrays that contain probes for 24,131 transcript probes. RESULTS: A total of 48 different samples (including duplicates) grown in multiple laboratories under different conditions were analyzed and pairwise comparisons were performed in all groups. Hierarchical clustering showed that blinded duplicates were correctly identified as the closest related samples. hESC lines clustered together irrespective of the laboratory in which they were maintained. hESCs could be readily distinguished from embryoid bodies (EB) differentiated from them and the karyotypically abnormal hESC line BG01V. The embryonal carcinoma (EC) line NTera2 is a useful model for evaluating characteristics of hESCs. Expression of subsets of individual genes was validated by comparing with published databases, MPSS (Massively Parallel Signature Sequencing) libraries, and parallel analysis by microarray and RT-PCR. CONCLUSION: we show that Illumina's bead array platform is a reliable, reproducible and robust method for developing base global profiles of cells and identifying similarities and differences in large number of samples

Safety and immunogenicity of AS03B adjuvanted split virion versus non-adjuvanted whole virion H1N1 influenza vaccine in UK children aged 6 months-12 years: open label, randomised, parallel group, multicentre study

Objectives To compare the safety, reactogenicity, and immunogenicity of an adjuvanted split virion H1N1 vaccine and a non-adjuvanted whole virion vaccine used in the pandemic immunisation programme in the United Kingdom

Single-crosslink microscopy in a biopolymer network dissects local elasticity from molecular fluctuations

Polymer networks are fundamental from cellular biology to plastics technology but their intrinsic inhomogeneity is masked by the usual ensemble-averaged measurements. Here, we construct direct maps of crosslinks-symbolic depiction of spatially-distributed elements highlighting their physical features and the relationships between them-in an actin network. We selectively label crosslinks with fluorescent markers, track their thermal fluctuations, and characterize the local elasticity and cross-correlations between crosslinks. Such maps display massive heterogeneity, reveal abundant anticorrelations, and may contribute to address how local responses scale up to produce macroscopic elasticity. Single-crosslink microscopy offers a general, microscopic framework to better understand crosslinked molecular networks in undeformed or strained states