Mycobacterium tuberculosis type VII secretion system effectors differentially impact the ESCRT endomembrane damage response

Mycobacterium tuberculosis causes tuberculosis, which kills more people than any other infection. M. tuberculosis grows in macrophages, cells that specialize in engulfing and degrading microorganisms. Like many intracellular pathogens, in order to cause disease, M. tuberculosis damages the membrane-bound compartment (phagosome) in which it is enclosed after macrophage uptake. Recent work showed that when chemicals damage this type of intracellular compartment, cells rapidly detect and repair the damage, using machinery called the endosomal sorting complex required for transport (ESCRT). Therefore, we hypothesized that ESCRT might also respond to pathogen-induced damage. At the same time, our previous work showed that the EsxG-EsxH heterodimer of M. tuberculosis can inhibit ESCRT, raising the possibility that M. tuberculosis impairs this host response. Here, we show that ESCRT is recruited to damaged M. tuberculosis phagosomes and that EsxG-EsxH undermines ESCRT-mediated endomembrane repair. Thus, our studies demonstrate a battle between host and pathogen over endomembrane integrity.Intracellular pathogens have varied strategies to breach the endolysosomal barrier so that they can deliver effectors to the host cytosol, access nutrients, replicate in the cytoplasm, and avoid degradation in the lysosome. In the case of Mycobacterium tuberculosis, the bacterium perforates the phagosomal membrane shortly after being taken up by macrophages. Phagosomal damage depends upon the mycobacterial ESX-1 type VII secretion system (T7SS). Sterile insults, such as silica crystals or membranolytic peptides, can also disrupt phagosomal and endolysosomal membranes. Recent work revealed that the host endosomal sorting complex required for transport (ESCRT) machinery rapidly responds to sterile endolysosomal damage and promotes membrane repair. We hypothesized that ESCRTs might also respond to pathogen-induced phagosomal damage and that M. tuberculosis could impair this host response. Indeed, we found that ESCRT-III proteins were recruited to M. tuberculosis phagosomes in an ESX-1-dependent manner. We previously demonstrated that the mycobacterial effectors EsxG/TB9.8 and EsxH/TB10.4, both secreted by the ESX-3 T7SS, can inhibit ESCRT-dependent trafficking of receptors to the lysosome. Here, we additionally show that ESCRT-III recruitment to sites of endolysosomal damage is antagonized by EsxG and EsxH, both within the context of M. tuberculosis infection and sterile injury. Moreover, EsxG and EsxH themselves respond within minutes to membrane damage in a manner that is independent of calcium and ESCRT-III recruitment. Thus, our study reveals that T7SS effectors and ESCRT participate in a series of measures and countermeasures for control of phagosome integrity

Data Handling and Analysis for the 1971 Corn Blight Watch Experiment

In the summer of 1971 the Laboratory for Applications of Remote Sensing at Purdue University conducted a pioneering experiment in the application of remote sensing technology to a large scale earth resource surveillance problem. In 1970 a plant disease called the Southern Corn Leaf Blight infected large areas of corn production in the United States and it is estimated that total national corn production pas reduced by 15% due to this disease. In the winter of 1970, an experiment was designed in cooperation with the U.S. Department of Agriculture, NASA, and the University of Michigan Institute of Science and Technology\u27s Willow Run Laboratories for the summer of 1971 to study the feasibility of detecting and mapping this disease using remote sensing techniques. In this paper, the Corn Blight Watch Experiment will be described from the point of view of data handling, data analysis and interpretation procedures used

Anderson Transition in Disordered Graphene

We use the regularized kernel polynomial method (RKPM) to numerically study the effect disorder on a single layer of graphene. This accurate numerical method enables us to study very large lattices with millions of sites, and hence is almost free of finite size errors. Within this approach, both weak and strong disorder regimes are handled on the same footing. We study the tight-binding model with on-site disorder, on the honeycomb lattice. We find that in the weak disorder regime, the Dirac fermions remain extended and their velocities decrease as the disorder strength is increased. However, if the disorder is strong enough, there will be a {\em mobility edge} separating {\em localized states around the Fermi point}, from the remaining extended states. This is in contrast to the scaling theory of localization which predicts that all states are localized in two-dimensions (2D).Comment: 4 page

Segregation by thermal diffusion in granular shear flows

Segregation by thermal diffusion of an intruder immersed in a sheared granular gas is analyzed from the (inelastic) Boltzmann equation. Segregation is induced by the presence of a temperature gradient orthogonal to the shear flow plane and parallel to gravity. We show that, like in analogous systems without shear, the segregation criterion yields a transition between upwards segregation and downwards segregation. The form of the phase diagrams is illustrated in detail showing that they depend sensitively on the value of gravity relative to the thermal gradient. Two specific situations are considered: i) absence of gravity, and ii) homogeneous temperature. We find that both mechanisms (upwards and downwards segregation) are stronger and more clearly separated when compared with segregation criteria in systems without shear.Comment: 8 figures. To appear in J. Stat. Mec

How my clinical placement in Australia helped me to become the clinician I am today

In the global society in which we live the graduate speech-language pathologist needs to be prepared for working with a culturally diverse client group and for the possibility that they may work in a country other than the one in which they trained. International clinical placement opportunities are a common method for many Australian speech language pathology programs to prepare students for an international career and for working with a culturally diverse client group. There have been many reported benefits for students taking part in these placements. But what are the benefits for overseas students who participate in a placement in Australia? This clinical insights article asked five clinicians who had trained in the UK and who had completed a placement in Australia during their training to reflect on this experience. They reported many benefits both personally and professionally. They felt that their Australian placement experience prepared them to work with a culturally diverse client group and shaped who they areas clinicians. There were also additional benefits for the service in which they now worked

Flexible Lipid Bilayers in Implicit Solvent

A minimalist simulation model for lipid bilayers is presented. Each lipid is represented by a flexible chain of beads in implicit solvent. The hydrophobic effect is mimicked through an intermolecular pair potential localized at the ``water''/hydrocarbon tail interface. This potential guarantees realistic interfacial tensions for lipids in a bilayer geometry. Lipids self assemble into bilayer structures that display fluidity and elastic properties consistent with experimental model membrane systems. Varying molecular flexibility allows for tuning of elastic moduli and area/molecule over a range of values seen in experimental systems.Comment: 5 pages, 5 figure

Physical characteristics underpinning repetitive lunging in fencing

Given the repetitive demand to execute lunging and changes in direction within fencing, the ability to sustain these at maximal capacity is fundamental to performance. The aim of this study was threefold. Firstly to provide normative values for this variable referred to as repeat lunge ability (RLA) and secondly to identify the physical characteristics that underpin it. Thirdly, was to establish if a cause and effect relationship existed by training the associated characteristics. Assessment of lower body power, reactive strength, speed, change of direction speed (CODS) and a sport specific RLA were conducted on senior and junior elite male fencers (n = 36). Fencers were on average (± SD) 18.9 ± 3.2 years of age, 174.35 ± 10.42 cm tall, 70.67 ± 7.35 kg in mass, and 8.5 ± 4.2 years fencing experience. The RLA test had average work times of 16.03 s ± 1.40 and demonstrated "large" to "very large" associations with all tested variables, but in particular CODS (r = .70) and standing broad jump (SBJ; r = -68). Through linear regression analysis, these also provided a two-predictor model accounting for 61% of the common variance associated with RLA. A cause and effect relationship with SBJ and CODS was confirmed by the training group, where RLA performance in these fencers improved from 15.80 ± 1.07 s to 14.90 ± 0.86 s, with the magnitude of change reported as "moderate" (ES = 0.93). Concurrent improvements were also noted in both SBJ (216.86 cm ± 17.15 vs. 221.71 ± 17.59 cm) and CODS (4.44 ± 0.29 s s. 4.31 ± 0.09 s) and while differences were only significant in SBJ, magnitudes of change were classed as "small" (ES = 0.28) and "moderate" (ES = 0.61)respectively. In conclusion, to improve RLA strength and conditioning coaches should focus on improving lower-body power and reactive strength, noting that jump training and plyometrics designed to enhance horizontal propulsion may be most effective, and translate to improvement in CODS also

Vectorial Loading of Processive Motor Proteins: Implementing a Landscape Picture

Individual processive molecular motors, of which conventional kinesin is the most studied quantitatively, move along polar molecular tracks and, by exerting a force ${\bm F} = (F_x,F_y,F_z)$ on a tether, drag cellular cargoes, {\em in vivo}, or spherical beads, {\em in vitro}, taking up to hundreds of nanometer-scale steps. From observations of velocities and the dispersion of displacements with time, under measured forces and controlled fuel supply (typically ATP), one may hope to obtain insight into the molecular motions undergone in the individual steps. In the simplest situation, the load force ${\bm F}$ may be regarded as a scalar resisting force, $F_x < 0$, acting parallel to the track: however, experiments, originally by Gittes {\em et al.} (1996), have imposed perpendicular (or vertical) loads, $F_z > 0$, while more recently Block and coworkers (2002, 2003) and Carter and Cross (2005) have studied {\em assisting} (or reverse) loads, $F_x > 0$, and also sideways (or transverse) loads $F_y \neq 0$

A programmable two-qubit quantum processor in silicon

With qubit measurement and control fidelities above the threshold of fault-tolerance, much attention is moving towards the daunting task of scaling up the number of physical qubits to the large numbers needed for fault tolerant quantum computing. Here, quantum dot based spin qubits may offer significant advantages due to their potential for high densities, all-electrical operation, and integration onto an industrial platform. In this system, the initialisation, readout, single- and two-qubit gates have been demonstrated in various qubit representations. However, as seen with other small scale quantum computer demonstrations, combining these elements leads to new challenges involving qubit crosstalk, state leakage, calibration, and control hardware which provide invaluable insight towards scaling up. Here we address these challenges and demonstrate a programmable two-qubit quantum processor in silicon by performing both the Deutsch-Josza and the Grover search algorithms. In addition, we characterise the entanglement in our processor through quantum state tomography of Bell states measuring state fidelities between 85-89% and concurrences between 73-80%. These results pave the way for larger scale quantum computers using spins confined to quantum dots

Metformin reduces airway glucose permeability and hyperglycaemia-induced Staphylococcus aureus load independently of effects on blood glucose

Background Diabetes is a risk factor for respiratory infection, and hyperglycaemia is associated with increased glucose in airway surface liquid and risk of Staphylococcus aureus infection. Objectives To investigate whether elevation of basolateral/blood glucose concentration promotes airway Staphylococcus aureus growth and whether pretreatment with the antidiabetic drug metformin affects this relationship. Methods Human airway epithelial cells grown at air–liquid interface (±18 h pre-treatment, 30 μM–1 mM metformin) were inoculated with 5×105 colony-forming units (CFU)/cm2 S aureus 8325-4 or JE2 or Pseudomonas aeruginosa PA01 on the apical surface and incubated for 7 h. Wild-type C57BL/6 or db/db (leptin receptor-deficient) mice, 6–10 weeks old, were treated with intraperitoneal phosphate-buffered saline or 40 mg/kg metformin for 2 days before intranasal inoculation with 1×107 CFU S aureus. Mice were culled 24 h after infection and bronchoalveolar lavage fluid collected. Results Apical S aureus growth increased with basolateral glucose concentration in an in vitro airway epithelia–bacteria co-culture model. S aureus reduced transepithelial electrical resistance (RT) and increased paracellular glucose flux. Metformin inhibited the glucose-induced growth of S aureus, increased RT and decreased glucose flux. Diabetic (db/db) mice infected with S aureus exhibited a higher bacterial load in their airways than control mice after 2 days and metformin treatment reversed this effect. Metformin did not decrease blood glucose but reduced paracellular flux across ex vivo murine tracheas. Conclusions Hyperglycaemia promotes respiratory S aureus infection, and metformin modifies glucose flux across the airway epithelium to limit hyperglycaemia-induced bacterial growth. Metformin might, therefore, be of additional benefit in the prevention and treatment of respiratory infection