Signature-tagged transposon mutagenesis studies demonstrate the dynamic nature of cecal colonization of 2-week-old chickens by Campylobacter jejuni.

We have constructed plasmids to be used for in vitro signature-tagged mutagenesis (STM) of Campylobacter jejuni and used these to generate STM libraries in three different strains. Statistical analysis of the transposon insertion sites in the C. jejuni NCTC 11168 chromosome and the plasmids of strain 81-176 indicated that their distribution was not uniform. Visual inspection of the distribution suggested that deviation from uniformity was not due to preferential integration of the transposon into a limited number of hot spots but rather that there was a bias towards insertions around the origin. We screened pools of mutants from the STM libraries for their ability to colonize the ceca of 2-week-old chickens harboring a standardized gut flora. We observed high-frequency random loss of colonization proficient mutants. When cohoused birds were individually inoculated with different tagged mutants, random loss of colonization-proficient mutants was similarly observed, as was extensive bird-to-bird transmission of mutants. This indicates that the nature of campylobacter colonization in chickens is complex and dynamic, and we hypothesize that bottlenecks in the colonization process and between-bird transmission account for these observations.

Metabolite and transcriptome analysis of Campylobacter jejuni in vitro growth reveals a stationary-phase physiological switch.

Campylobacter jejuni is a prevalent cause of food-borne diarrhoeal illness in humans. Understanding of the physiological and metabolic capabilities of the organism is limited. We report a detailed analysis of the C. jejuni growth cycle in batch culture. Combined transcriptomic, phenotypic and metabolic analysis demonstrates a highly dynamic 'stationary phase', characterized by a peak in motility, numerous gene expression changes and substrate switching, despite transcript changes that indicate a metabolic downshift upon the onset of stationary phase. Video tracking of bacterial motility identifies peak activity during stationary phase. Amino acid analysis of culture supernatants shows a preferential order of amino acid utilization. Proton NMR (1H-NMR) highlights an acetate switch mechanism whereby bacteria change from acetate excretion to acetate uptake, most probably in response to depletion of other substrates. Acetate production requires pta (Cj0688) and ackA (Cj0689), although the acs homologue (Cj1537c) is not required. Insertion mutants in Cj0688 and Cj0689 maintain viability less well during the stationary and decline phases of the growth cycle than wild-type C. jejuni, suggesting that these genes, and the acetate pathway, are important for survival.

Transposon mutagenesis in a hyper-invasive clinical isolate of Campylobacter jejuni reveals a number of genes with potential roles in invasion.

Transposon mutagenesis has been applied to a hyper-invasive clinical isolate of Campylobacter jejuni, 01/51. A random transposon mutant library was screened in an in vitro assay of invasion and 26 mutants with a significant reduction in invasion were identified. Given that the invasion potential of C. jejuni is relatively poor compared to other enteric pathogens, the use of a hyper-invasive strain was advantageous as it greatly facilitated the identification of mutants with reduced invasion. The location of the transposon insertion in 23 of these mutants has been determined; all but three of the insertions are in genes also present in the genome-sequenced strain NCTC 11168. Eight of the mutants contain transposon insertions in one region of the genome (approximately 14 kb), which when compared with the genome of NCTC 11168 overlaps with one of the previously reported plasticity regions and is likely to be involved in genomic variation between strains. Further characterization of one of the mutants within this region has identified a gene that might be involved in adhesion to host cells.

An extended computational model of the circulatory system for designing ventricular assist devices.

An extended computational model of the circulatory system has been developed to predict blood flow in the presence of ventricular assist devices (VADs). A novel VAD, placed in the descending aorta, intended to offload the left ventricle (LV) and augment renal perfusion is being studied. For this application, a better understanding of the global hemodynamic response of the VAD, in essence an electrically driven pump, and the cardiovascular system is necessary. To meet this need, a model has been established as a nonlinear, lumped-parameter electrical analog, and simulated results under different states [healthy, congestive heart failure (CHF), and postinsertion of VAD] are presented. The systemic circulation is separated into five compartments and the descending aorta is composed of three components to accurately yield the system response of each section before and after the insertion of the VAD. Delays in valve closing time and blood inertia in the aorta were introduced to deliver a more realistic model. Pump governing equations and optimization are based on fundamental theories of turbomachines and can serve as a practical initial design point for rotary blood pumps. The model's results closely mimic established parameters for the circulatory system and confirm the feasibility of the intra-aortic VAD concept. This computational model can be linked with models of the pump motor to provide a valuable tool for innovative VAD design.

Ground vibration generated by trains in underground tunnels

A popular method used to reduce vibration transmitted from underground railways into nearby buildings is floating-slab track, whereby a concrete slab supporting the two rails is mounted on rubber bearings or steel springs to isolate it from the tunnel invert. This paper adds a track model to a previously developed three-dimensional tunnel model in order to assess the effectiveness of floating-slab track. A slab beam coupled to the tunnel in the wavenumber domain, with the slab bearings represented by an elastic layer, is examined first. A second beam representing the two rails together is then coupled to the slab, and axle masses representing a train are added to the rail beam. Power-spectral densities and RMS levels of soil vibration due to random roughness-displacement excitation between the masses and the rail beam are calculated. Analytical techniques are used to minimise the computational requirements of the model. The results demonstrate the inadequacy of simple mass-spring and Winkler-beam models with rigid foundations for the assessment of the vibration-isolation performance of railway track. They suggest that the achievable insertion loss is modest and that floating the track slab may in fact cause increased transmission of vibration under certain conditions. © 2006 Elsevier Ltd. All rights reserved.

Inserting innovations in-service

Military platforms have exceptionally long lifecycles and given the state of defense budgets there is a significant trend in sustaining the operational capability of legacy platforms for much greater periods than originally designed. In the context of through-life management, one of the key questions is how to manage the flow of technology for platform modernization during the in-service phase of the lifecycle? Inserting technological innovations in-service is achieved through technology insertion processes. Technology insertion is the pre-eminent activity for both maintaining and enhancing the functional capability of a platform especially given the likely changes in future military operations, the pace of change in technology and with the increasing focus on lifecycle cost reduction. This chapter provides an introduction to technology insertion together with an overview of the key issues that practitioners are faced with. As an aid to planning technology insertion projects, a decision-support framework is presented. © 2010 Springer-Verlag Berlin Heidelberg.

The effect of a twin tunnel on the propagation of ground-borne vibration from an underground railway

Accurate predictions of ground-borne vibration levels in the vicinity of an underground railway are greatly sought after in modern urban centres. Yet the complexity involved in simulating the underground environment means that it is necessary to make simplifying assumptions about this system. One such commonly made assumption is to ignore the effects of neighbouring tunnels, despite the fact that many underground railway lines consist of twin-bored tunnels, one for the outbound direction and one for the inbound direction. This paper presents a unique model for two tunnels embedded in a homogeneous, elastic fullspace. Each of these tunnels is subject to both known, dynamic train forces and dynamic cavity forces. The net forces acting on the tunnels are written as the sum of those tractions acting on the invert of a single tunnel, and those tractions that represent the motion induced by the neighbouring tunnel. By apportioning the tractions in this way, the vibration response of a two-tunnel system is written as a linear combination of displacement fields produced by a single-tunnel system. Using Fourier decomposition, forces are partitioned into symmetric and antisymmetric modenumber components to minimise computation times. The significance of the interactions between two tunnels is quantified by calculating the insertion gains, in both the vertical and horizontal directions, that result from the existence of a second tunnel. The insertion-gain results are shown to be localised and highly dependent on frequency, tunnel orientation and tunnel thickness. At some locations, the magnitude of these insertion gains is greater than 20 dB. This demonstrates that a high degree of inaccuracy exists in any surface vibration prediction model that includes only one of the two tunnels. This novel two-tunnel solution represents a significant contribution to the existing body of research into vibration from underground railways, as it shows that the second tunnel has a significant influence on the accuracy of vibration predictions for underground railways. © 2011 Elsevier Ltd. All rights reserved.

Wideband tunable, high-power, graphene mode-locked ultrafast lasers

Ultrafast passively mode-locked lasers with spectral tuning capability and high output power have widespread applications in biomedical research, spectroscopy and telecommunications [1,2]. Currently, the dominant technology is based on semiconductor saturable absorber mirrors (SESAMs) [2,3]. However, these typically have a narrow tuning range, and require complex fabrication and packaging [2,3]. A simple, cost-effective alternative is to use Single Wall Carbon Nanotubes (SWNTs) [4,10] and Graphene [10,14]. Wide-band operation is possible using SWNTs with a wide diameter distribution [5,10]. However, SWNTs not in resonance are not used and may contribute to unwanted insertion losses [10]. The linear dispersion of the Dirac electrons in graphene offers an ideal solution for wideband ultrafast pulse generation [10,15]. © 2011 IEEE.

Significant improvements in optical power budgets of real-time optical OFDM PON systems.

Based on a comprehensive theoretical optical orthogonal frequency division multiplexing (OOFDM) system model rigorously verified by comparing numerical results with end-to-end real-time experimental measurements at 11.25Gb/s, detailed explorations are undertaken, for the first time, of the impacts of various physical factors on the OOFDM system performance over directly modulated DFB laser (DML)-based, intensity modulation and direct detection (IMDD), single-mode fibre (SMF) systems without in-line optical amplification and chromatic dispersion compensation. It is shown that the low extinction ratio (ER) of the DML modulated OOFDM signal is the predominant factor limiting the maximum achievable optical power budget, and the subcarrier intermixing effect associated with square-law photon detection in the receiver reduces the optical power budget by at least 1dB. Results also indicate that, immediately after the DML in the transmitter, the insertion of a 0.02nm bandwidth optical Gaussian bandpass filter with a 0.01nm wavelength offset with respect to the optical carrier wavelength can enhance the OOFDM signal ER by approximately 1.24dB, thus resulting in a 7dB optical power budget improvement at a total channel BER of 1 × 10(-3).