Construction and evaluation of plasmid vectors optimized for constitutive and regulated gene expression in Burkholderia cepacia complex isolates

Genetic studies with Burkholderia cepacia complex isolates are hampered by the limited availability of cloning vectors and by the inherent resistance of these isolates to the most common antibiotics used for genetic selection. Also, some of the promoters widely employed for gene expression in Escherichia coli are inefficient in B. cepacia. In this study, we have utilized the backbone of the vector pME6000, a derivative of the pBBR1 plasmid that was originally isolated from Bordetella bronchiseptica, to construct a set of vectors useful for gene expression in B. cepacia. These vectors contain either the constitutive promoter of the S7 ribosomal protein gene from Burkholderia sp. strain LB400 or the arabinose-inducible P(BAD) promoter from E. coli. Promoter sequences were placed immediately upstream of multiple cloning sites in combination with the minimal sequence of pME6000 required for plasmid maintenance and mobilization. The functionality of both vectors was assessed by cloning the enhanced green fluorescent protein gene (e-gfp) and determining the levels of enhanced green fluorescent protein expression and fluorescence emission for a variety of clinical and environmental isolates of the B. cepacia complex. We also demonstrate that B. cepacia carrying these constructs can readily be detected intracellularly by fluorescence microscopy following the infection of Acanthamoeba polyphaga.

Optomechanical cooling with generalized interferometers

The fields in multiple-pass interferometers, such as the Fabry-Pérot cavity, exhibit great sensitivity not only to the presence but also to the motion of any scattering object within the optical path. We consider the general case of an interferometer comprising an arbitrary configuration of generic beam splitters and calculate the velocity-dependent radiation field and the light force exerted on a moving scatterer. We find that a simple configuration, in which the scatterer interacts with an optical resonator from which it is spatially separated, can enhance the optomechanical friction by several orders of magnitude.

Exciton-mediated photothermal cooling in GaAs membranes

Cooling of the mechanical motion of a GaAs nano-membrane using the photothermal effect mediated by excitons was recently demonstrated by some of the authors (Usami et al 2012 Nature Phys. 8 168) and provides a clear example of the use of thermal forces to cool down mechanical motion. Here, we report on a single-free-parameter theoretical model to explain the results of this experiment which matches the experimental data remarkably well.

Scattering theory of cooling and heating in optomechanical systems

We present a one-dimensional scattering theory which enables us to describe a wealth of effects arising from the coupling of the motional degree of freedom of scatterers to the electromagnetic field. Multiple scattering to all orders is taken into account. The theory is applied to describe the scheme of a Fabry-Perot resonator with one of its mirrors moving. The friction force, as well as the diffusion, acting on the moving mirror is derived. In the limit of a small reflection coefficient, the same model provides for the description of the mechanical effect of light on an atom moving in front of a mirror.

Cavity cooling of atoms: Within and without a cavity

We compare the efficiencies of two optical cooling schemes, where a single particle is either inside or outside an optical cavity, under experimentally-realisable conditions. We evaluate the cooling forces using the general solution of a transfer matrix method for a moving scatterer inside a general one-dimensional system composed of immobile optical elements. Assuming the same atomic saturation parameter, we find that the two cooling schemes provide cooling forces and equilibrium temperatures of comparable magnitude.

Atom cooling using the dipole force of a single retroreflected laser beam

We present a mechanism for cooling atoms by a laser beam reflected from a single mirror. The cooling relies on the dipole force and thus in principle applies to arbitrary refractive particles including atoms, molecules, or dielectric spheres. Friction and equilibrium temperatures are derived by an analytic perturbative approach. Finally, semiclassical Monte-Carlo simulations are performed to validate the analytic results.

Optical Cooling of Atoms in Microtraps by Time-Delayed Reflection

We present a theoretical analysis of a novel scheme for optical cooling of particles that does not in principle require a closed optical transition. A tightly confined laser beam interacting with a trapped particle experiences a phase shift, which upon reflection from a mirror or resonant microstructure produces a time-delayed optical potential for the particle. This leads to a nonconservative force and friction. A quantum model of the system is presented and analyzed in the semiclassical limit.

Optical Cooling Using the Dipole Force

The term `laser cooling' is applied to the use of optical means to cool the motional energies of either atoms and molecules, or micromirrors. In the literature, these two strands are kept largely separate; both, however suffer from severe limitations. Laser cooling of atoms and molecules largely relies on the internal level structure of the species being cooled. As a result, only a small number of elements and a tiny number of molecules can be cooled this way. In the case of micromirrors, the problem lies in the engineering of micromirrors that need to satisfy a large number of constraints---these include a high mechanical Q-factor, high reflectivity and very good optical quality, weak coupling to the substrate, etc.---in order to enable efficient cooling. During the course of this thesis, I will draw these two sides of laser cooling closer together by means of a single, generically applicable scattering theory that can be used to explain the interaction between light and matter at a very general level. I use this `transfer matrix' formalism to explore the use of the retarded dipole--dipole interaction as a means of both enhancing the efficiency of micromirror cooling systems and rendering the laser cooling of atoms and molecules less species selective. In particular, I identify the `external cavity cooling' mechanism, whereby the use of an optical memory in the form of a resonant element (such as a cavity), outside which the object to be cooled sits, can potentially lead to the construction of fully integrated optomechanical systems and even two-dimensional arrays of translationally cold atoms, molecules or even micromirrors.

Optimized bit level architectures for IIR filtering

Optimized circuits for implementing high-performance bit-parallel IIR filters are presented. Circuits constructed mainly from simple carry save adders and based on most-significant-bit (MSB) first arithmetic are described. Two methods resulting in systems which are 100% efficient in that they are capable of sampling data every cycle are presented. In the first approach the basic circuit is modified so that the level of pipelining used is compatible with the small, but fixed, latency associated with the computation in question. This is achieved through insertion of pipeline delays (half latches) on every second row of cells. This produces an area-efficient solution in which the throughput rate is determined by a critical path of 76 gate delays. A second approach combines the MSB first arithmetic methods with the scattered look-ahead methods. Important design issues are addressed, including wordlength truncation, overflow detection, and saturation.

Detached-Eddy Simulation of a Louver-Cooling Scheme for Turbine Blades

The performance of a louver-cooling scheme on a flat plate was analyzed using a detached-eddy-simulation turbulence model. It was assumed that the louver-cooling scheme was tested in a wind tunnel with the mainstream flow velocity of 20 m/s, equivalent to a Reynolds number of 16,200, based on the jet diameter. Turbulence closure was achieved by a realizable k-e-based detached-eddy-simulation turbulence model. Solutions of two blowing ratios of 0.5 and 1 were successfully obtained by running parallel on 16 nodes on a computer cluster. The flowfields were found to be highly unsteady and oscillatory in nature, with the maximum fluctuation of the adiabatic effectiveness as high as 15% of the time-averaged value. It is shown that the fluctuations in the adiabatic effectiveness are mainly caused by the spanwise fluctuation of the coolant jet and the unsteady vortical structures created by the interaction of the jet and the mainstream.

Unsteady Simulations of a Film Cooling Flow from an Inclined Cylindrical Jet

Film cooling is extensively used to provide protection against the severe thermal environment in gas turbine engines. Most of the computational studies on film cooling flow have been done using steady Reynolds-averaged Navier–Stokes calculation procedures. However, the flowfield associated with a jet in a crossflow is highly unsteady and complex with different types of vortical structures. In this paper, a computational investigation about the unsteady phenomena of a jet in a crossflow is performed using detached eddy simulation. Detailed computation of a single row of 35 deg round holes on a flat plate has been obtained for a 1.0 blowing ratio and a 2.0 density ratio. First, time-step size, grid resolution, and computational domain tests for an unsteady simulation have been conducted. Comparison between the results of unsteady Reynolds-averaged Navier–Stokes calculation, detached eddy simulation, and large eddy simulation is also performed. Comparison of the time-averaged detached eddy simulation prediction with the measured film-cooling effectiveness shows that the detached eddy simulation prediction is reasonable. From present detached eddy simulations, the influential coherent vortical structures of a film cooling flow can be seen. The unsteady physics of jet in a crossflow interactions and a jet liftoff in film cooling flows have been explained.