msbB deletion confers acute sensitivity to CO2 in Salmonella enterica serovar Typhimurium that can be suppressed by a loss-of-function mutation in zwf

<p>Abstract</p> <p>Background</p> <p>Pathogens tolerate stress conditions that include low pH, oxidative stress, high salt and high temperature in order to survive inside and outside their hosts. Lipopolysaccharide (LPS), which forms the outer-leaflet of the outer membrane in Gram-negative bacteria, acts as a permeability barrier. The lipid A moiety of LPS anchors it to the outer membrane bilayer. The MsbB enzyme myristoylates the lipid A precursor and loss of this enzyme, in <it>Salmonella</it>, is correlated with reduced virulence and severe growth defects that can both be compensated with extragenic suppressor mutations.</p> <p>Results</p> <p>We report here that <it>msbB </it>(or <it>msbB somA</it>) <it>Salmonella </it>are highly sensitive to physiological CO₂(5%), resulting in a 3-log reduction in plating efficiency. Under these conditions, <it>msbB Salmonella </it>form long filaments, bulge and lyse. These bacteria are also sensitive to acidic pH and high osmolarity. Although CO₂acidifies LB broth media, buffering LB to pH 7.5 did not restore growth of <it>msbB </it>mutants in CO₂, indicating that the CO₂-induced growth defects are not due to the effect of CO₂on the pH of the media. A transposon insertion in the glucose metabolism gene <it>zwf </it>compensates for the CO₂sensitivity of <it>msbB Salmonella</it>. The <it>msbB zwf </it>mutants grow on agar, or in broth, in the presence of 5% CO₂. In addition, <it>msbB zwf </it>strains show improved growth in low pH or high osmolarity media compared to the single <it>msbB </it>mutant.</p> <p>Conclusion</p> <p>These results demonstrate that <it>msbB </it>confers acute sensitivity to CO₂, acidic pH, and high osmolarity. Disruption of <it>zwf </it>in <it>msbB </it>mutants restores growth in 5% CO₂and results in improved growth in acidic media or in media with high osmolarity. These results add to a growing list of phenotypes caused by <it>msbB </it>and mutations that suppress specific growth defects.</p

LUVMI: an innovative payload for the sampling of volatiles at the Lunar poles

The ISECG identifies one of the first exploration steps as in situ investigations of the moon or asteroids. Europe is developing payload concepts for drilling and sample analysis, a contribution to a 250kg rover as well as for sample return. To achieve these missions, ESA depends on international partnerships. Such missions will be seldom, expensive and the drill/sample site selected will be based on observations from orbit not calibrated with ground truth data. Many of the international science community’s objectives can be met at lower cost, or the chances of mission success improved and the quality of the science increased by making use of an innovative, low mass, mobile robotic payload following the LEAG recommendations. LUVMI provides a smart, low mass, innovative, modular mobile payload comprising surface and subsurface sensing with an in-situ sampling technology capable of depth-resolved extraction of volatiles, combined with a volatile analyser (mass spectrometer) capable of identifying the chemical composition of the most important volatiles. This will allow LUVMI to: traverse the lunar surface prospecting for volatiles; sample subsurface up to a depth of 10 cm (with a goal of 20 cm); extract water and other loosely bound volatiles; identify the chemical species extracted; access and sample permanently shadowed regions (PSR). The main innovation of LUVMI is to develop an in situ sampling technology capable of depth-resolved extraction of volatiles, and then to package within this tool, the analyser itself, so as to maximise transfer efficiency and minimise sample handling and its attendant mass requirements and risk of sample alteration. By building on national, EC and ESA funded research and developments, this project will develop to TRL6 instruments that together form a smart modular mobile payload that could be flight ready in 2020. The LUVMI sampling instrument will be tested in a highly representative environment including thermal, vacuum and regolith simulant and the integrated payload demonstrated in a representative environment

Integrated assessment of marine biodiversity status using a prototype indicator-based assessment tool

Integrated assessment of the status of marine biodiversity is and has been problematic compared to, for example, assessments of eutrophication and contamination status, mostly as a consequence of the fact that monitoring of marine habitats, communities and species is expensive, often collected at an incorrect spatial scale and/or poorly integrated with existing marine environmental monitoring efforts. The objective of this Method Paper is to introduce and describe a simple tool for integrated assessment of biodiversity status based on the HELCOM Biodiversity Assessment Tool (BEAT), where interim biodiversity indicators are grouped by themes: broad-scale habitats, communities, and species as well as supporting non-biodiversity indicators. Further, we report the application of an initial indicator-based assessment of biodiversity status of Danish marine waters where we have tentatively classified the biodiversity status of Danish marine waters. The biodiversity status was in no areas classified as “unaffected by human activities.” In all the 22 assessment areas, the status was classified as either “moderately affected by human activities” or “significantly affected by human activities.” Spatial variations in the biodiversity status were in general related to the eutrophication status as well as fishing pressure

Risk stratification in patients with acute chest pain using three high-sensitivity cardiac troponin assays

Aims Several high-sensitivity cardiac troponin (hs-cTn) assays have recently been developed. It is unknown which hs-cTn provides the most accurate prognostic information and to what extent early changes in hs-cTn predict mortality. Methods and results In a prospective, international multicentre study, cTn was simultaneously measured with three novel [high-sensitivity cardiac Troponin T (hs-cTnT), Roche Diagnostics; hs-cTnI, Beckman-Coulter; hs-cTnI, Siemens] and a conventional assay (cTnT, Roche Diagnostics) in a blinded fashion in 1117 unselected patients with acute chest pain. Patients were followed up 2 years regarding mortality. Eighty-two (7.3%) patients died during the follow-up. The 2-year prognostic accuracy of hs-cTn was most accurate for hs-cTnT [area under the receivers operating characteristic curve (AUC) 0.78 (95% CI: 0.73-0.83) and outperformed both hs-cTnI (Beckman-Coulter, 0.71 (95% CI: 0.65-0.77; P = 0.001 for comparison), hs-cTnI (Siemens) 0.70 (95% CI: 0.64-0.76; P < 0.001 for comparison)] and cTnT 0.67 (95% CI: 0.61-0.74; P < 0.001 for comparison). Absolute changes of hs-cTnT were more accurate than relative changes in predicting mortality, but inferior to presentation values of hs-cTnT. Combining changes of hs-cTnT within the first 6 h with their presentation values did not further improve prognostic accuracy. Similar results were obtained for both hs-cTnI assays regarding the incremental value of changes. Hs-cTn concentrations remained predictors of death in clinically challenging subgroups such as patients with pre-existing coronary artery disease, impaired renal function, and patients older than 75 years. Conclusion High-sensitivity cardiac Troponin T is more accurate than hs-cTnI in the prediction of long-term mortality. Changes of hs-cTn do not seem to further improve risk stratification beyond initial presentation value

The status of GEO 600

The GEO 600 laser interferometer with 600m armlength is part of a worldwide network of gravitational wave detectors. GEO 600 is unique in having advanced multiple pendulum suspensions with a monolithic last stage and in employing a signal recycled optical design. This paper describes the recent commissioning of the interferometer and its operation in signal recycled mode

Dominant Mutations in GRHL3 Cause Van der Woude Syndrome and Disrupt Oral Periderm Development

Mutations in interferon regulatory factor 6 (IRF6) account for ∼70% of cases of Van der Woude syndrome (VWS), the most common syndromic form of cleft lip and palate. In 8 of 45 VWS-affected families lacking a mutation in IRF6, we found coding mutations in grainyhead-like 3 (GRHL3). According to a zebrafish-based assay, the disease-associated GRHL3 mutations abrogated periderm development and were consistent with a dominant-negative effect, in contrast to haploinsufficiency seen in most VWS cases caused by IRF6 mutations. In mouse, all embryos lacking Grhl3 exhibited abnormal oral periderm and 17% developed a cleft palate. Analysis of the oral phenotype of double heterozygote (Irf6+/−;Grhl3+/−) murine embryos failed to detect epistasis between the two genes, suggesting that they function in separate but convergent pathways during palatogenesis. Taken together, our data demonstrated that mutations in two genes, IRF6 and GRHL3, can lead to nearly identical phenotypes of orofacial cleft. They supported the hypotheses that both genes are essential for the presence of a functional oral periderm and that failure of this process contributes to VWS

Ab initio van der Waals interactions in simulations of water alter structure from mainly tetrahedral to high-density-like

The structure of liquid water at ambient conditions is studied in ab initio molecular dynamics simulations using van der Waals (vdW) density-functional theory, i.e. using the new exchange-correlation functionals optPBE-vdW and vdW-DF2. Inclusion of the more isotropic vdW interactions counteracts highly directional hydrogen-bonds, which are enhanced by standard functionals. This brings about a softening of the microscopic structure of water, as seen from the broadening of angular distribution functions and, in particular, from the much lower and broader first peak in the oxygen-oxygen pair-correlation function (PCF), indicating loss of structure in the outer solvation shells. In combination with softer non-local correlation terms, as in the new parameterization of vdW-DF, inclusion of vdW interactions is shown to shift the balance of resulting structures from open tetrahedral to more close-packed. The resulting O-O PCF shows some resemblance with experiment for high-density water (A. K. Soper and M. A. Ricci, Phys. Rev. Lett., 84:2881, 2000), but not directly with experiment for ambient water. However, an O-O PCF consisting of a linear combination of 70% from vdW-DF2 and 30% from experiment on low-density liquid water reproduces near-quantitatively the experimental O-O PCF for ambient water, indicating consistency with a two-liquid model with fluctuations between high- and low-density regions

Effect of thrombolysis with alteplase within 6 h of acute ischaemic stroke on long-term outcomes (the third International Stroke Trial [IST-3]): 18-month follow-up of a randomised controlled trial

SummaryBackgroundFew data are available from randomised trials about the effect of thrombolysis with alteplase on long-term functional outcome in patients who have had acute ischaemic stroke and no trial has reported effects on health-related quality of life. A secondary objective of the third International Stroke Trial (IST-3) was to assess the effect of thrombolysis on such outcomes at 18 months.MethodsIn this open-label, international, multicentre, randomised, controlled trial, 3035 patients with ischaemic stroke from 12 countries were randomly allocated within 6 h of onset via a secure central system to either intravenous alteplase (0·9 mg/kg; n=1515) plus standard care or standard care alone (control; n=1520). 2348 patients were scheduled for 18-month follow-up. For our main analysis, survivors were assessed at 18 months with the Oxford handicap scale (OHS; the primary outcome was the adjusted odds of OHS score 0–2). We also used the EuroQoL (EQ) instrument and asked questions about overall functioning and living circumstances. We analysed the OHS and the five EQ domains by ordinal logistic regression and calculated the mean difference between treatment groups in EQ utility index and visual analogue scale score. Analyses were adjusted for key baseline prognostic factors. This study is registered with controlled-trials.com, number ISRCTN25765518.FindingsAt 18 months, 408 (34·9%) of 1169 patients in the alteplase group versus 414 (35·1%) of 1179 in the control group had died (p=0·85). 391 (35·0%) of 1117 patients versus 352 (31·4%) of 1122 had an OHS score of 0–2 (adjusted odds ratio [OR] 1·28, 95% CI 1·03–1·57; p=0·024). Treatment was associated with a favourable shift in the distribution of OHS grades (adjusted common OR 1·30, 95% CI 1·10–1·55; p=0·002). Alteplase treatment was associated with significantly higher overall self-reported health (adjusted mean difference in EQ utility index 0·060; p=0·019). The differences between the groups in visual analogue scale score and the proportion living at home were not significant.InterpretationIST-3 provides evidence that thrombolysis with intravenous alteplase for acute ischaemic stroke does not affect survival, but does lead to statistically significant, clinically relevant improvements in functional outcome and health-related quality of life that are sustained for at least 18 months.FundingUK Medical Research Council, Health Foundation UK, Stroke Association UK, Research Council of Norway, AFA Insurances Sweden, Swedish Heart Lung Fund, The Foundation of Marianne and Marcus Wallenberg, Polish Ministry of Science and Education, the Australian Heart Foundation, Australian National Health and Medical Research Council, Swiss National Research Foundation, Swiss Heart Foundation, Assessorato alla Sanita (Regione dell'Umbria, Italy), and Danube University

Longitudinal Diffusion Tensor Imaging Resembles Patterns of Pathology Progression in Behavioral Variant Frontotemporal Dementia (bvFTD)

Objective: Recently, the characteristic longitudinal distribution pattern of the underlying phosphorylated TDP-43 (pTDP-43) pathology in the behavioral variant of frontotemporal dementia (bvFTD) excluding Pick's disease (PiD) across specific brain regions was described. The aim of the present study was to investigate whether in vivo investigations of bvFTD patients by use of diffusion tensor imaging (DTI) were consistent with these proposed patterns of progression. Methods: Sixty-two bvFTD patients and 47 controls underwent DTI in a multicenter study design. Of these, 49 bvFTD patients and 34 controls had a follow-up scan after ~12 months. Cross-sectional and longitudinal alterations were assessed by a two-fold analysis, i.e., voxelwise comparison of fractional anisotropy (FA) maps and a tract of interest-based (TOI) approach, which identifies tract structures that could be assigned to brain regions associated with disease progression. Results: Whole brain-based spatial statistics showed white matter alterations predominantly in the frontal lobes cross-sectionally and longitudinally. The TOIs of bvFTD neuroimaging stages 1 and 2 (uncinate fascicle—bvFTD pattern I; corticostriatal pathway—bvFTD pattern II) showed highly significant differences between bvFTD patients and controls. The corticospinal tract-associated TOI (bvFTD pattern III) did not differ between groups, whereas the differences in the optic radiation (bvFTD pattern IV) reached significance. The findings in the corticospinal tract were due to a “dichotomous” behavior of FA changes there. Conclusion: Longitudinal TOI analysis demonstrated a pattern of white matter pathways alterations consistent with patterns of pTDP-43 pathology