High Content Phenotypic Cell-Based Visual Screen Identifies Mycobacterium tuberculosis Acyltrehalose-Containing Glycolipids Involved in Phagosome Remodeling

The ability of the tubercle bacillus to arrest phagosome maturation is considered one major mechanism that allows its survival within host macrophages. To identify mycobacterial genes involved in this process, we developed a high throughput phenotypic cell-based assay enabling individual sub-cellular analysis of over 11,000 Mycobacterium tuberculosis mutants. This very stringent assay makes use of fluorescent staining for intracellular acidic compartments, and automated confocal microscopy to quantitatively determine the intracellular localization of M. tuberculosis. We characterised the ten mutants that traffic most frequently into acidified compartments early after phagocytosis, suggesting that they had lost their ability to arrest phagosomal maturation. Molecular analysis of these mutants revealed mainly disruptions in genes involved in cell envelope biogenesis (fadD28), the ESX-1 secretion system (espL/Rv3880), molybdopterin biosynthesis (moaC1 and moaD1), as well as in genes from a novel locus, Rv1503c-Rv1506c. Most interestingly, the mutants in Rv1503c and Rv1506c were perturbed in the biosynthesis of acyltrehalose-containing glycolipids. Our results suggest that such glycolipids indeed play a critical role in the early intracellular fate of the tubercle bacillus. The unbiased approach developed here can be easily adapted for functional genomics study of intracellular pathogens, together with focused discovery of new anti-microbials

Trace elements in hemodialysis patients: a systematic review and meta-analysis

<p>Abstract</p> <p>Background</p> <p>Hemodialysis patients are at risk for deficiency of essential trace elements and excess of toxic trace elements, both of which can affect health. We conducted a systematic review to summarize existing literature on trace element status in hemodialysis patients.</p> <p>Methods</p> <p>All studies which reported relevant data for chronic hemodialysis patients and a healthy control population were eligible, regardless of language or publication status. We included studies which measured at least one of the following elements in whole blood, serum, or plasma: antimony, arsenic, boron, cadmium, chromium, cobalt, copper, fluorine, iodine, lead, manganese, mercury, molybdenum, nickel, selenium, tellurium, thallium, vanadium, and zinc. We calculated differences between hemodialysis patients and controls using the differences in mean trace element level, divided by the pooled standard deviation.</p> <p>Results</p> <p>We identified 128 eligible studies. Available data suggested that levels of cadmium, chromium, copper, lead, and vanadium were higher and that levels of selenium, zinc and manganese were lower in hemodialysis patients, compared with controls. Pooled standard mean differences exceeded 0.8 standard deviation units (a large difference) higher than controls for cadmium, chromium, vanadium, and lower than controls for selenium, zinc, and manganese. No studies reported data on antimony, iodine, tellurium, and thallium concentrations.</p> <p>Conclusion</p> <p>Average blood levels of biologically important trace elements were substantially different in hemodialysis patients, compared with healthy controls. Since both deficiency and excess of trace elements are potentially harmful yet amenable to therapy, the hypothesis that trace element status influences the risk of adverse clinical outcomes is worthy of investigation.</p

On the bridging mechanism in vibration controlled dispensing of pharmaceutical powders from a micro hopper

Accurate batch dispensing of pharmaceutical powders, on the scale of hundreds of microns, in small doses is a challenging task. A novel dispensing technique has been developed by Yang et al. using high-frequency vibration to control powder flow out of a narrow hopper. This method removes the need for mechanical valves because the powder, very quickly, forms a bridge-like structure across the passive outlet preventing outflow. Activation of the vibration has been found to destabilise the bridging structure enabling the powder to flow, when vibration stops the bridge structure quickly rebuilds and dispensing stops. In this work the Discrete Element Method (DEM) was used to simulate this novel dispensing control method in order to identify the internal mechanism that allows the flow to be controlled so precisely. A simulated conical hopper was filled with particles then oscillated vertically at high frequency (≈. 10. kHz), amplitude and frequency were scaled from the experimental system. Two orifice sizes, a variety of DEM parameters and two vibration modes were simulated. The parametric study of DEM parameters was based around a case that provided flow rates within a factor of 2 of the experimental flow rates. Dispensing after vibration was found to stop very quickly as in experiments. Visualisation of internal structures during fill, vibration and immediately after vibration revealed a central mass of slow moving particles floating above the nozzle outlet. When the vibration stops the central mass of particles drops into contact with the walls and quickly plugs the flow. © 2013 Elsevier B.V.status: publishe

Combined aerodynamic and electrostatic atomization of dielectric liquid jets

The electrical and atomization performance of a plane-plane charge injection atomizer using a dielectric liquid, and operating at pump pressures ranging from 15 to 35 bar corresponding to injection velocities of up to 50m/s, is explored via low current electrical measurements, spray imaging and phase Doppler anemometry (PDA). The work is aimed at understanding the contribution of electrostatic charging relevant to typical higher pressure fuel injection systems such as those employed in the aeronautical, automotive and marine sectors. Results show that mean specific charge increases with injection velocity significantly. The effect of electrostatic charge is advantageous at the 15-35 bar range and an arithmetic mean diameter D10 as low as 0.2d is achievable in the spray core and lower still in the periphery where d is the orifice diameter. Using the data available from this higher pressure system and from previous high Reynolds number systems [1], the promotion of primary atomization has been analyzed by examining the effect that charge has on liquid jet surface and liquid jet bulk instability. The results suggest that for the low charge density Qv~2C/m3 cases under consideration here, a significant increase in primary atomization is observed due to a combination of electrical and aerodynamic forces acting on the jet surface, attributed to the significantly higher jet Weber number (Wej) when compared to low injection pressure cases. Analysis of Sauter mean diameter (SMD) results show that for jets with elevated specific charge density of the order QV~6C/m3, the jet creates droplets that a conventional turbulent jet would, but with a significantly lower power requirement. This suggests that “turbulent” primary atomization, the turbulence being induced by electrical forces, may be achieved under injection pressures that would produce laminar jets

Small Changes in Gene Expression of Targeted Osmoregulatory Genes When Exposing Marine and Freshwater Threespine Stickleback (Gasterosteus aculeatus) to Abrupt Salinity Transfers

Salinity is one of the key factors that affects metabolism, survival and distribution of fish species, as all fish osmoregulate and euryhaline fish maintain osmotic differences between their extracellular fluid and either freshwater or seawater. The threespine stickleback (Gasterosteus aculeatus) is a euryhaline species with populations in both marine and freshwater environments, where the physiological and genomic basis for salinity tolerance adaptation is not fully understood. Therefore, our main objective in this study was to investigate gene expression of three targeted osmoregulatory genes (Na+/K+-ATPase (ATPA13), cystic fibrosis transmembrane regulator (CFTR) and a voltage gated potassium channel gene (KCNH4) and one stress related heat shock protein gene (HSP70)) in gill tissue from marine and freshwater populations when exposed to non-native salinity for periods ranging from five minutes to three weeks. Overall, the targeted genes showed highly plastic expression profiles, in addition the expression of ATP1A3 was slightly higher in saltwater adapted fish and KCNH4 and HSP70 had slightly higher expression in freshwater. As no pronounced changes were observed in the expression profiles of the targeted genes, this indicates that the osmoregulatory apparatuses of both the marine and landlocked freshwater stickleback population have not been environmentally canalized, but are able to respond plastically to abrupt salinity challenges