A rhodanine agent active against non-replicating intracellular Mycobacterium avium subspecies paratuberculosis.

BACKGROUND: Antibiotic therapy targeting chronic mycobacterial disease is often ineffective due to problems with the emergence of drug resistance and non-replicating persistent intracellular antibiotic resistant phenotypes. Strategies which include agents able to enhance host cell killing mechanisms could represent an alternative to conventional methods with the potential for host clearance if active against dormant phenotypes. Investigations of agents with potential activity against non-replicating mycobacteria however are restricted due to a need for assays that can assess bacterial viability without having to culture. RESULTS: This study describes the development and use of a pre16S ribosomal gene RNA/DNA ratio viability assay which is independent of the need for culture, supported by a novel thin layer accelerated mycobacterial colony forming method for determining viability and culturability of MAP in intracellular environments. We describe the use of these tools to demonstrate intracellular killing activity of a novel rhodanine agent (D157070) against the intracellular pathogen Mycobacterium avium subspecies paratuberculosis (MAP) and show that the culturability of MAP decreases relative to its viability on intracellular entry suggesting the induction of a non-culturable phenotype. We further demonstrate that D157070, although having no direct activity against the culturability of extracellular MAP, can bind to cultured MAP cells and has significant influence on the MAP transcriptome, particularly with respect of delta(L )associated genes. D157070 is shown to be taken up by bovine and human cells and able to enhance host cell killing, as measured by significant decreases in both culturability and viability of intracellular MAP. CONCLUSIONS: This work suggests that pre16srRNA gene ratios represent a viable method for studying MAP viability. In addition, the rhodanine agent D157070 tested is non-toxic and enhances cell killing activity against both growing and latent MAP phenotypes

Measurement of the lowest millimetre-wave transition frequency of the CH radical

The CH radical offers a sensitive way to test the hypothesis that fundamental constants measured on earth may differ from those observed in other parts of the universe. The starting point for such a comparison is to have accurate laboratory frequencies. Here we measure the frequency of the lowest millimetre-wave transition of CH, near 535 GHz, with an accuracy of 0.6 kHz. This improves the uncertainty by roughly two orders of magnitude over previous determinations and opens the way for sensitive new tests of varying constants.Comment: 5 pages, 5 figure

Prospects for measuring the electric dipole moment of the electron using electrically trapped polar molecules

Heavy polar molecules can be used to measure the electric dipole moment of the electron, which is a sensitive probe of physics beyond the Standard Model. The value is determined by measuring the precession of the molecule's spin in a plane perpendicular to an applied electric field. The longer this precession evolves coherently, the higher the precision of the measurement. For molecules in a trap, this coherence time could be very long indeed. We evaluate the sensitivity of an experiment where neutral molecules are trapped electrically, and compare this to an equivalent measurement in a molecular beam. We consider the use of a Stark decelerator to load the trap from a supersonic source, and calculate the deceleration efficiency for YbF molecules in both strong-field seeking and weak-field seeking states. With a 1s holding time in the trap, the statistical sensitivity could be ten times higher than it is in the beam experiment, and this could improve by a further factor of five if the trap can be loaded from a source of larger emittance. We study some effects due to field inhomogeneity in the trap and find that rotation of the electric field direction, leading to an inhomogeneous geometric phase shift, is the primary obstacle to a sensitive trap-based measurement.Comment: 22 pages, 7 figures, prepared for Faraday Discussion 14

Stochastic multi-channel lock-in detection

High-precision measurements benefit from lock-in detection of small signals. Here we discuss the extension of lock-in detection to many channels, using mutually orthogonal modulation waveforms, and show how the the choice of waveforms affects the information content of the signal. We also consider how well the detection scheme rejects noise, both random and correlated. We address the particular difficulty of rejecting a background drift that makes a reproducible offset in the output signal and we show how a systematic error can be avoided by changing the waveforms between runs and averaging over many runs. These advances made possible a recent measurement of the electron's electric dipole moment.Comment: 11 pages, 3 figure

Bar graph monitor

Bar graph monitor of pulse position modulation telemetry ground station equipment for sounding rocket

Probing the electron EDM with cold molecules

We present progress towards a new measurement of the electron electric dipole moment using a cold supersonic beam of YbF molecules. Data are currently being taken with a sensitivity of $10^{-27}\textrm{e.cm}/\sqrt{\textrm{day}}$. We therefore expect to make an improvement over the Tl experiment of Commins' group, which currently gives the most precise result. We discuss the systematic and statistical errors and comment on the future prospect of making a measurement at the level of $10^{-29}\textrm{e.cm}/\sqrt{\textrm{day}}$.Comment: 8 pages, 6 figures, proceedings of ICAP 200

A robust floating nanoammeter

A circuit capable of measuring nanoampere currents while floating at voltages up to at least 25kV is described. The circuit relays its output to ground potential via an optical fiber. We particularly emphasize the design and construction techniques which allow robust operation in the presence of high voltage spikes and discharges.Comment: 5 pages, 2 figure

Spectral Properties and Lifetimes of Neutral Spin-1/2-Fermions in a Magnetic Guide

We investigate the resonant motion of neutral spin-1/2-fermions in a magnetic guide. A wealth of unitary and anti-unitary symmetries is revealed in particular giving rise to a two-fold degeneracy of the energy levels. To compute the energies and decay widths of a large number of resonances the complex scaling method is employed. We discuss the dependence of the lifetimes on the angular momentum of the resonance states. In this context the existence of so-called quasi-bound states is shown. In order to approximately calculate the resonance energies of such states a radial Schr\"odinger equation is derived which improves the well-known adiabatic approximation. The effects of an additionally applied homogeneous Ioffe field on the resonance energies and decay widths are also considered. The results are applied to the case of the $^6\text{Li}$ atom in the $F=1/2$ hyperfine ground state.Comment: accepted for publication in PR

Prospects for the measurement of the electron electric dipole moment using YbF

We discuss an experiment underway at Imperial College London to measure the permanent electric dipole moment (EDM) of the electron using a molecular beam of YbF. We describe the measurement method, which uses a combination of laser and radiofrequency resonance techniques to detect the spin precession of the YbF molecule in a strong electric field. We pay particular attention to the analysis scheme and explore some of the possible systematic effects which might mimic the EDM signal. Finally, we describe technical improvements which should increase the sensitivity by more than an order of magnitude over the current experimental limit.Comment: 6 pages, 2 figure