Eliminating helper phage from phage display

Phage display technology involves the display of proteins or peptides, as coat protein fusions, on the surface of a phage or phagemid particles. Using standard technology, helper phage are essential for the replication and assembly of phagemid particles, during library production and biopanning. We have eliminated the need to add helper phage by using 'bacterial packaging cell lines' that provide the same functions. These cell lines contain M13-based helper plasmids that express phage packaging proteins which assemble phagemid particles as efficiently as helper phage, but without helper phage contamination. This results in genetically pure phagemid particle preparations. Furthermore, by using constructs differing in the form of gene 3 that they contain, we have shown that the display, from a single library, can be modulated between monovalent (phagemid-like) and multivalent display (phage-like) without any further engineering. These packaging cells eliminate the use of helper phage from phagemid-based selection protocols; reducing the amount of technical preparation, facilitating automation, optimizing selections by matching display levels to diversity, and effectively using the packaged phagemid particles as means to transfer genetic information at an efficiency approaching 100%

The value of habitats of conservation importance to climate change mitigation in the UK

The input of PS contributes to the following UKRI-funded projects: DEVIL (NE/M021327/1) and Soils-R-GRREAT (NE/P019455/1). The contributions of RHF, GMB, AH and RBB were funded by RSPB. Thanks to Olly Watts and Phil Grice and three anonymous reviewers for comments on earlier versions of this paper, their comments immeasurably improved it. Thanks also to Ruth Davies, Mel Coath, Vicki Swales, Arfon Williams, Tom Lancaster, Andrew Midgely, Phillip Carson, Jonathan Cryer, Adam Barnett, Paul Morling, Pat Thompson, Beatriz Luraschi, Kate Jennings and Jonathon Bell who helped frame the question addressed here and shape the ‘storymap’ based on this work, available at https://arcg.is/098uiD.Peer reviewedPostprin

Hyperuniversality of Fully Anisotropic Three-Dimensional Ising Model

For the fully anisotropic simple-cubic Ising lattice, the critical finite-size scaling amplitudes of both the spin-spin and energy-energy inverse correlation lengths and the singular part of the reduced free-energy density are calculated by the transfer-matrix method and a finite-size scaling for cyclic L x L x oo clusters with L=3 and 4. Analysis of the data obtained shows that the ratios and the directional geometric means of above amplitudes are universal.Comment: RevTeX 3.0, 24 pages, 2 figures upon request, accepted for publication in Phys. Rev.

Signal and Charge Transfer Efficiency of Few Electrons Clocked on Microscopic Superfluid Helium Channels

Electrons floating on the surface of liquid helium are possible spin-qubits for quantum information processing. Varying electric potentials are not expected to modify spin states, which allows their transport on helium using a charge-coupled device (CCD)-like array of underlying gates. This approach depends upon efficient inter-gate transfer of individual electrons. Measurements are presented here of the charge transfer efficiency (CTE) of few electrons clocked back and forth above a short microscopic CCD-like structure. A charge transfer efficiency of 0.99999992 is obtained for a clocking frequency of 800 kHz.Comment: 13 pages including 3 figure

Very High Energy gamma-ray observations of Mrk 501 using TACTIC imaging gamma-ray telescope during 2005-06

In this paper we report on the Markarian 501 results obtained during our TeV $\gamma$-ray observations from March 11 to May 12, 2005 and February 28 to May 7, 2006 for 112.5 hours with the TACTIC $\gamma$-ray telescope. During 2005 observations for 45.7 hours, the source was found to be in a low state and we have placed an upper limit of 4.62 $\times$ 10$^{-12}$ photons cm$^{-2}$ s$^{-1}$ at 3$\sigma$ level on the integrated TeV $\gamma$-ray flux above 1 TeV from the source direction. However, during the 2006 observations for 66.8h, detailed data analysis revealed the presence of a TeV $\gamma$-ray signal from the source with a statistical significance of 7.5$\sigma$ above $E_{\gamma}\geq$ 1 TeV. The time averaged differential energy spectrum of the source in the energy range 1-11 TeV is found to match well with the power law function of the form ($d\Phi/dE=f_0 E^{-\Gamma}$) with $f_0=(1.66\pm0.52)\times 10^{-11}cm^{-2}s^{-1}TeV^{-1}$ and $\Gamma=2.80\pm0.27$.Comment: 16 pages and 8 Figures Accepted for publication in the Journal of Physics

Multiwavelength Observations of a Flare from Markarian 501

We present multiwavelength observations of the BL Lacertae object Markarian 501 (Mrk 501) in 1997 between April 8 and April 19. Evidence of correlated variability is seen in very high energy (VHE, E > 350 GeV) gamma-ray observations taken with the Whipple Observatory gamma-ray telescope, data from the Oriented Scintillation Spectrometer Experiment of the Compton Gamma-Ray Observatory, and quicklook results from the All-Sky Monitor of the Rossi X-ray Timing Explorer while the Energetic Gamma-Ray Experiment Telescope did not detect Mrk 501. Short term optical correlations are not conclusive but the U-band flux observed with the 1.2m telescope of the Whipple Observatory was 10% higher than in March. The average energy output of Mrk 501 appears to peak in the 2 keV to 100 keV range suggesting an extension of the synchrotron emission to at least 100 keV, the highest observed in a blazar and ~100 times higher than that seen in the other TeV-emitting BL Lac object, Mrk 421. The VHE gamma-ray flux observed during this period is the highest ever detected from this object. The VHE gamma-ray energy output is somewhat lower than the 2-100 keV range but the variability amplitude is larger. The correlations seen here do not require relativistic beaming of the emission unless the VHE spectrum extends to >5 TeV.Comment: 10 pages, 2 figures, accepted for publication in ApJ Letter

Characteristics and Treatments of Patients with Peripheral Arterial Disease Referred to UK Vascular Clinics: Results of a Prospective Registry

BackgroundPeripheral arterial disease (PAD) is often associated with risk factors including cigarette smoking, hypertension and hypercholesterolaemia, and patients have a high risk of future vascular events. Good medical management results in improved outcomes and quality of life, but previous studies have documented sub-optimal treatment of risk factors. We assessed the management of cardiovascular risk factors in patients with PAD referred to specialist vascular clinics.MethodsThis was a prospective, protocol driven registry carried out in UK vascular clinics. Patients who were first-time referrals for evaluation of PAD were eligible if they had claudication plus ankle-brachial pressure index (ABPI) ≤0.9. Statistical associations between key demographic and treatment variables were explored using a chi-squared test.ResultsWe enrolled 473 patients from 23 sites. Mean age was 68 years (SD 10) and 66% were male. Mean estimated claudication distance was 100m, and ABPI was 0.74. Mean systolic blood pressure (SBP) was 155mmHg, and 42% had a SBP >160mmHg. Forty percent were current smokers and half had tried to give up in the prior 6 months, but there was no evidence of a systematic method of smoking cessation. Mean total cholesterol was 5.4 (SD1.2) mmol/l and 30% had levels >6mmol/l. Antiplatelet therapy had been given to 70% and statins to 44%. Prior CHD was present in 29% and these patients had significantly higher use of antiplatelet therapy, statins and ACE-inhibitors.ConclusionsIn spite of attempts to raise awareness about PAD as an important marker of cardiovascular risk, patients are still poorly treated prior to referral to a vascular clinic. In particular, the use of evidence-based treatments is sub-optimal, while hypertension and cigarette smoking are poorly managed. More work needs to be done to educate health professionals about the detection and optimal medical management of PAD

Parsec Scale Properties of Markarian 501

We present the results of a high angular resolution study of the BL Lac object Markarian 501 in the radio band. We consider data taken at 14 different epochs, ranging between 1.6 GHz and 22 GHz in frequency, and including new Space VLBI observations obtained on 2001 March 5 and 6 at 1.6 and 5 GHz. We study the kinematics of the parsec-scale jet and estimate its bulk velocity and orientation with respect to the line of sight. Limb brightened structure in the jet is clearly visible in our data and we discuss its possible origin in terms of velocity gradients in the jet. Quasi-simultaneous multi-wavelength observations allow us to map the spectral index distribution and to compare it to the jet morphology. Finally, we estimate the physical parameters of the parsec-scale jet.Comment: accepted for publication in ApJ; 24 pages with 17 figures (fig. 1 and fig. 2 available only as .jpg files

The Landscape of Reason: A Scheme for Representing Arguments Concerning Environmental, Health and Safety Effects of Chemical Weapons Disposal in the US

To reduce the risk of environmental contamination and honor an international treaty, chemical weapons stored at eight locales around the US are slated for destruction. Incineration is the main choice of a National Research Council committee directed by Congress to weigh the hazards of alternative destruction technologies, but many citizens\u27 groups remain unconvinced. The US Army, which must dispose of the dangerous chemicals, faces decisions about the choice of destruction technologies, as well as more specific questions concerning protection of environment, safety and public health once the technology choices are made. Based on more than 200 individual interviews and 40 focus groups held in communities near where the weapons are stored, this paper illustrates an argumentation scheme for representing the underlying reasons for varying positions in the conflict over technology choices. The argumentation scheme is effective in representing qualitative interview data concerning the complex and dynamic environmental perspectives of diverse regional and national constituencies