The VgrG Proteins Are "à la Carte" Delivery Systems for Bacterial Type VI Effectors

The bacterial type VI secretion system (T6SS) is a supra-molecular complex akin to bacteriophage tails, with VgrG proteins acting as a puncturing device. The Pseudomonas aeruginosa H1-T6SS has been extensively characterized. It is involved in bacterial killing and in the delivery of three toxins, Tse1–3. Here, we demonstrate the independent contribution of the three H1-T6SS co-regulated vgrG genes, vgrG1abc, to bacterial killing. A putative toxin is encoded in the vicinity of each vgrG gene, supporting the concept of specific VgrG/toxin couples. In this respect, VgrG1c is involved in the delivery of an Rhs protein, RhsP1. The RhsP1 C terminus carries a toxic activity, from which the producing bacterium is protected by a cognate immunity. Similarly, VgrG1a-dependent toxicity is associated with the PA0093 gene encoding a two-domain protein with a putative toxin domain (Toxin_61) at the C terminus. Finally, VgrG1b-dependent killing is detectable upon complementation of a triple vgrG1abc mutant. The VgrG1b-dependent killing is mediated by PA0099, which presents the characteristics of the superfamily nuclease 2 toxin members. Overall, these data develop the concept that VgrGs are indispensable components for the specific delivery of effectors. Several additional vgrG genes are encoded on the P. aeruginosa genome and are not linked genetically to other T6SS genes. A closer inspection of these clusters reveals that they also encode putative toxins. Overall, these associations further support the notion of an original form of secretion system, in which VgrG acts as the carrier

Influence of a 12.8-km military load carriage activity on lower limb gait mechanics and muscle activity

This is the author accepted manuscript. The final version is available from the publisher via the DOI in this record.The high stress fracture occurrence in military populations has been associated with frequent load carriage activities. This study aimed to assess the influence of load carriage and of completing a load carriage training activity on gait characteristics. Thirty-two Royal Marine recruits completed a 12.8-km load carriage activity as part of their military training. Data were collected during walking in military boots, pre and post-activity, with and without the additional load (35.5 kg). Ground contact time, lower limb sagittal plane kinematics and kinetics, and electromyographic variables were obtained for each condition. When carrying load, there was increased ground contact time, increased joint flexion and joint moments, and increased plantar flexor and knee extensor muscle activity. Post-activity, there were no changes to kinematic variables, knee extensor moments were reduced, and there was evidence of plantar flexor muscle fatigue. The observed gait changes may be associated with stress fracture development. Practitioner Summary: This study identified gait changes due to load carriage and after a military load carriage training activity. Such activities are associated with lower limb stress fractures. A pre-post study design was used. Gait mechanics changed to a greater extent when carrying load, than after completion of the activity when assessed without load.This work was part-funded by the Institute of Naval Medicine

Facet recovery and light emission from GaN/InGaN/GaN core-shell structures grown by metal organic vapour phase epitaxy on etched GaN nanorod arrays

The use of etched nanorods from a planar template as a growth scaffold for a highly regular GaN/InGaN/GaN core-shell structure is demonstrated. The recovery of m-plane non-polar facets from etched high-aspect-ratio GaN nanorods is studied with and without the introduction of a hydrogen silsesquioxane passivation layer at the bottom of the etched nanorod arrays. This layer successfully prevented c-plane growth between the nanorods, resulting in vertical nanorod sidewalls (∼89.8°) and a more regular height distribution than re-growth on unpassivated nanorods. The height variation on passivated nanorods is solely determined by the uniformity of nanorod diameter, which degrades with increased growth duration. Facet-dependent indium incorporation of GaN/InGaN/GaN core-shell layers regrown onto the etched nanorods is observed by high-resolution cathodoluminescence imaging. Sharp features corresponding to diffracted wave-guide modes in angle-resolved photoluminescence measurements are evidence of the uniformity of the full core-shell structure grown on ordered etched nanorods

Altered forefoot function following a military training activity

This is the author accepted manuscript. The final version is available from Elsevier via the DOI in this recordBackground Injury rates are high in populations that regularly undertake weight-bearing physical activity, particularly military populations. Military training activities, that often include load carriage, have been associated with lower limb injury occurrence, specifically stress fractures. Research question Recent work identified plantar loading variables as risk factors for lower limb stress fractures in Royal Marines recruits that were assessed during barefoot running. This study aimed to quantify how those plantar loading variables changed in Royal Marines recruits following a prolonged military load carriage activity, to further understand potential mechanisms for lower limb stress fractures. Methods Bilateral, synchronised plantar pressure and lower limb kinematic data were recorded during barefoot running at 3.6 m s-1 (±5%) pre- and post- a 12.8-km training activity (∼150 min). The training activity was completed with an average speed typical of walking (1.4 m.s-1), and 35.5 kg of additional load was carried throughout. Data were collected from 32 male Royal Marines recruits who completed the training activity in week-21 of the 32-week training programme. Plantar pressure variables and ankle dorsiflexion were compared between pre- and post-activity. Results Post-activity there was reduced loading under the forefoot and increased loading under the rearfoot and midfoot. There was no change in dorsiflexion touchdown angle, but an increase in peak dorsiflexion and range of motion post-activity. Significance The increased rearfoot loading, reduced forefoot loading and increased ankle dorsiflexion following a prolonged military load carriage activity suggest a reduced transfer of loading from the rearfoot to the forefoot during stance, which may have implications for the development of stress fractures, particularly of the metatarsals.Institute of Naval Medicin

Diversity and prevalence of type VI secretion system effectors in clinical Pseudomonas aeruginosa isolates

Pseudomonas aeruginosa is an opportunistic pathogen and a major driver of morbidity and mortality in people with Cystic Fibrosis (CF). The Type VI secretion system (T6SS) is a molecular nanomachine that translocates effectors across the bacterial membrane into target cells or the extracellular environment enabling intermicrobial interaction. P. aeruginosa encodes three T6SS clusters, the H1-, H2- and H3-T6SS, and numerous orphan islands. Genetic diversity of T6SS-associated effectors in P. aeruginosa has been noted in reference strains but has yet to be explored in clinical isolates. Here, we perform a comprehensive bioinformatic analysis of the pangenome and T6SS effector genes in 52 high-quality clinical P. aeruginosa genomes isolated from CF patients and housed in the Personalised Approach to P. aeruginosa strain repository. We confirm that the clinical CF isolate pangenome is open and principally made up of accessory and unique genes that may provide strain-specific advantages. We observed genetic variability in some effector/immunity encoding genes and show that several well-characterised vgrG and PAAR islands are absent from numerous isolates. Our analysis shows clear evidence of disruption to T6SS genomic loci through transposon, prophage, and mobile genetic element insertions. We identified an orphan vgrG island in P. aeruginosa strain PAK and five clinical isolates using in silico analysis which we denote vgrG7, predicting a gene within this cluster to encode a Tle2 lipase family effector. Close comparison of T6SS loci in clinical isolates compared to reference P. aeruginosa strain PAO1 revealed the presence of genes encoding eight new T6SS effectors with the following putative functions: cytidine deaminase, lipase, metallopeptidase, NADase, and pyocin. Finally, the prevalence of characterised and putative T6SS effectors were assessed in 532 publicly available P. aeruginosa genomes, which suggests the existence of accessory effectors. Our in silico study of the P. aeruginosa T6SS exposes a level of genetic diversity at T6SS genomic loci not seen to date within P. aeruginosa, particularly in CF isolates. As understanding the effector repertoire is key to identifying the targets of T6SSs and its efficacy, this comprehensive analysis provides a path for future experimental characterisation of these mediators of intermicrobial competition and host manipulation

Strain evolution in GaN Nanowires: from free-surface objects to coalesced templates

Top-down fabricated GaN nanowires, 250 nm in diameter and with various heights, have been used to experimentally determine the evolution of strain along the vertical direction of 1-dimensional objects. X-ray diffraction and photoluminescence techniques have been used to obtain the strain profile inside the nanowires from their base to their top facet for both initial compressive and tensile strains. The relaxation behaviors derived from optical and structural characterizations perfectly match the numerical results of calculations based on a continuous media approach. By monitoring the elastic relaxation enabled by the lateral free-surfaces, the height from which the nanowires can be considered strain-free has been estimated. Based on this result, NWs sufficiently high to be strain-free have been coalesced to form a continuous GaN layer. X-ray diffraction, photoluminescence and cathodoluminescence clearly show that despite the initial strain-free nanowires template, the final GaN layer is strained

A narrow bimalleolar width is a risk factor for ankle inversion injury in male military recruits: A prospective study

This is the author accepted manuscript. The final version is available from the publisher via the DOI in this record.Background: Ankle inversion injuries are one of the most common and burdensome injuries in athletic populations. Research that prospectively identifies characteristics associated with this injury is lacking. This prospective study compared baseline anthropometric and biomechanical gait characteristics of military recruits who sustained an ankle inversion injury during training, with those who remained injury-free. Methods: Bilateral plantar pressure and three-dimensional lower limb kinematics were recorded in 1065 male, injury-free military recruits, during barefoot running. Injuries that occurred during the 32-week recruit training programme were subsequently recorded. Data were compared between recruits who sustained an ankle inversion injury during training (n=27) and a sample (n=120) of those who completed training injury-free. A logistic regression analysis was used to identify risk factors for this injury. Findings: A narrower bimalleolar width and an earlier peak pressure under the fifth metatarsal were predictors of ankle inversion injury. Those who sustained an ankle inversion injury also had a lower body mass, body mass index, and a smaller calf girth than those who completed training injury-free. Interpretation: Anthropometric and dynamic gait characteristics have been identified that may predispose recruits to an ankle inversion injury during Royal Marine recruit training, allowing identification of recruits at higher risk at the start of training

Calibrating whole building energy model: a case study using BEMS data

This paper describes a Calibration methodology which is specifically configured to best match actual building performance, based on a case study conducted to calibrate whole building energy model using Building Energy Management System (BEMS) measured data. It details the calibration approach which was designed to meet the specific characteristic of the spaces, systems and energy use in the pilot school building. Two calibration methods were developed; one is for electrical and the other is for thermal energy along with calibrated weather file. The result shows excellent correlation with the measured electricity and room air temperature and demonstrates the effectiveness of the methodology. Mean Bias Error (MBE) and Cumulative Variation of Root Mean Squared Error (CVRMSE) for electricity consumption is 6% and 14% respectively and -5 and 7% for air temperature

Giant Conductance Oscillations In Mesoscopic Andreev Interferometers

We analyze the electrical conductance $G(\phi)$ of a two-dimensional, phase coherent structure in contact with two superconductors, which is known to be an oscillatory function of the phase difference $\phi$ between the superconductors. It is predicted that for a metallic sample, the amplitude of oscillation is enhanced by placing a normal barrier at the interface and that, by tuning the strength of the barrier, can be orders of magnitude greater than values observed in recent experiments. Giant oscillations can also be obtained without a barrier, provided a crucial sum rule is broken. This can be achieved by disorder induced normal scattering. In the absence of zero phase inter-channel scattering, the conductance possesses a zero phase minimum.Comment: 4 pages of Revtex, 6 figures available on reques