The Aharonov-Bohm Effect in the Fractional Quantum Hall Regime

We have investigated experimentally resonant tunnelling through single-particle states formed around an antidot by a magnetic field, in the fractional quantum Hall regime. For 1/3 filling factor around the antidot, Aharonov-Bohm oscillations are observed with the same magnetic field period as in the integer quantum Hall regime. All our measurements are consistent with quasiparticles of fractional charge e*. However, the results are also consistent with particles of any charge (>= e*) as the system must rearrange every time the flux enclosed increases by h/e.Comment: Postscript, 4 pages, gzipped (350 kB

Inactivation Efficacy of Atmospheric Air Plasma and Airborne Acoustic Ultrasound Against Bacterial Bioflms

Bioflms are complex microbial communities that present serious contamination risks to our environment and health. In this study, atmospheric air plasma and airborne acoustic ultrasound technology were applied to inactivate Escherichia coli and Listeria innocua bioflms. Both technologies were efcient in controlling, or completely inactivating, the target bacterial bioflms. Viability and metabolic assays, along with microscopy analysis, revealed that atmospheric air plasma and airborne acoustic ultrasound damaged both the bacterial bioflm cells and its structural integrity. Scanning electron microscopy images highlighted the disruption of the bioflms and pore formation in bacterial cells exposed to both the plasma and acoustic treatments. Elevated reactive oxygen and nitrogen species in bacterial cells treated with atmospheric air plasma, demonstrated their primary role in the observed bacterial inactivation process. Our fndings provide potential antimicrobial strategies to combat bacterial bioflms in the food and healthcare sectors

Paradigm of biased PAR1 (protease-activated receptor-1) activation and inhibition in endothelial cells dissected by phosphoproteomics

Thrombin is the key serine protease of the coagulation cascade and mediates cellular responses by activation of PARs (protease-activated receptors). The predominant thrombin receptor is PAR1, and in endothelial cells (ECs), thrombin dynamically regulates a plethora of phosphorylation events. However, it has remained unclear whether thrombin signaling is exclusively mediated through PAR1. Furthermore, mechanistic insight into activation and inhibition of PAR1-mediated EC signaling is lacking. In addition, signaling networks of biased PAR1 activation after differential cleavage of the PAR1 N terminus have remained an unresolved issue. Here, we used a quantitative phosphoproteomics approach to show that classical and peptide activation of PAR1 induce highly similar signaling, that low thrombin concentrations initiate only limited phosphoregulation, and that the PAR1 inhibitors vorapaxar and parmodulin-2 demonstrate distinct antagonistic properties. Subsequent analysis of the thrombin-regulated phosphosites in the presence of PAR1 inhibitors revealed that biased activation of PAR1 is not solely linked to a specific G-protein downstream of PAR1. In addition, we showed that only the canonical thrombin PAR1 tethered ligand induces extensive early phosphoregulation in ECs. Our study provides detailed insight in the signaling mechanisms downstream of PAR1. Our data demonstrate that thrombin-induced EC phosphoregulation is mediated exclusively through PAR1, that thrombin and thrombin-tethered ligand peptide induce similar phosphoregulation, and that only canonical PAR1 cleavage by thrombin generates a tethered ligand that potently induces early signaling. Furthermore, platelet PAR1 inhibitors directly affect EC signaling, indicating that it will be a challenge to design a PAR1 antagonist that will target only those pathways responsible for tissue pathology

Changes in the capacity of visual working memory in 5- to 10-year-olds

Using the Luck and Vogel change detection paradigm, we sought to investigate the capacity of visual working memory in 5-, 7-, and 10-year-olds. We found that performance on the task improved significantly with age and also obtained evidence that the capacity of visual working memory approximately doubles between 5 and 10 years of age, where it reaches adult levels of approximately three to four items

Spectroscopic evidence for the binary nature of AM CVn

We analysed archival spectroscopic data of AM CVn taken with the William Herschel Telescope in 1996. In the literature two orbital periods for AM CVn are proposed. A clear S-wave in the HeI 4471, 4387 and 4143 \AA lines is revealed when the spectra are folded on the 1029 s period. No signature of this S-wave is seen when folded on 1051 s. Doppler tomography of the line profiles shows a clear signature of the hotspot. Using this we can constrain the value of K_2 to lie between 210 and 280 km/s. Our work confirms the binary nature of AM CVn beyond any doubt, establishes 1028.73 s as the true orbital period and supports the interpretation of AM CVn as a permanent superhump system.Comment: Accepted by MNRA

Solution structure of the Hop TPR2A domain and investigation of target druggability by NMR, biochemical and in silico approaches

Heat shock protein 90 (Hsp90) is a molecular chaperone that plays an important role in tumour biology by promoting the stabilisation and activity of oncogenic ‘client’ proteins. Inhibition of Hsp90 by small-molecule drugs, acting via its ATP hydrolysis site, has shown promise as a molecularly targeted cancer therapy. Owing to the importance of Hop and other tetratricopeptide repeat (TPR)-containing cochaperones in regulating Hsp90 activity, the Hsp90-TPR domain interface is an alternative site for inhibitors, which could result in effects distinct from ATP site binders. The TPR binding site of Hsp90 cochaperones includes a shallow, positively charged groove that poses a significant challenge for druggability. Herein, we report the apo, solution-state structure of Hop TPR2A which enables this target for NMR-based screening approaches. We have designed prototype TPR ligands that mimic key native ‘carboxylate clamp’ interactions between Hsp90 and its TPR cochaperones and show that they block binding between Hop TPR2A and the Hsp90 C-terminal MEEVD peptide. We confirm direct TPR-binding of these ligands by mapping 1H–15N HSQC chemical shift perturbations to our new NMR structure. Our work provides a novel structure, a thorough assessment of druggability and robust screening approaches that may offer a potential route, albeit difficult, to address the chemically challenging nature of the Hop TPR2A target, with relevance to other TPR domain interactors

Enzymatic activities and functional interdependencies of Bacillus subtilis lipoteichoic acid synthesis enzymes

Lipoteichoic acid (LTA) is an important cell wall polymer in Gram-positive bacteria. The enzyme responsible for polyglycerolphosphate LTA synthesis is LtaS, first described in Staphylococcus aureus. Four LtaS orthologues, LtaSBS, YfnI, YqgS and YvgJ, are present in Bacillus subtilis. Using an in vitro enzyme assay, we determined that all four proteins are Mn2+-dependent metal enzymes that use phosphatidylglycerol as a substrate. We show that LtaSBS, YfnI and YqgS can produce polymers, suggesting that these three proteins are bona-fide LTA synthases while YvgJ functions as an LTA primase, as indicated by the accumulation of a GroP-Glc2-DAG glycolipid. Western blot analysis of LTA produced by ltaSBS, yfnI, yqgS and yvgJ single, triple and the quadruple mutant, showed that LTA production was only abolished in the quadruple and the YvgJ-only expressing mutant. B. subtilis strains expressing YfnI in the absence of LtaSBS produced LTA of retarded mobility, presumably caused by an increase in chain length as suggested by a structural analysis of purified LTA. Taken together, the presented results indicate that the mere presence or absence of LTA cannot account for cell division and sporulation defects observed in the absence of individual enzymes and revealed an unexpected enzymatic interdependency of LtaS-type proteins in B. subtilis

Damage evolution in freeze cast metal/ceramic composites exhibiting lamellar microstructures

The damage evolution in a single domain aluminium/alumina freeze-cast composite has been examined using 3D X-ray computed tomography (CT). A single domain was extracted and loaded incrementally at an orientation of 45° to the lamellae, with the damage being assessed after each of eight compressive loading steps. Prior to loading, significant damage was observed at the metal-ceramic interface – this is thought to have formed during machining and can be ascribed to weak interfacial bonding associated with the Cu coating applied to the ceramic preform prior to metal infiltration. Further interfacial damage was seen to initiate after loading to 170MPa and to develop with each subsequent load step. Damage was also observed in the ceramic lamellae, with a series of parallel cracks forming across the alumina, perpendicular to the domain orientation.These sets of parallel, intra-lamellae cracks were closely spaced, but initiated independently, with coalescenceonly occurring at higher loads. Both the interfacial and intra-lamellae cracking initiated after loading to 170MPa,with the intra-lamellae cracks propagating into the metal matrix after loading to 240MPa. The cracks in the ceramic lamellae were found to form and develop independent of the interfacial cracks, with discrete crackpaths and morphologies being observed in each case. Despite this, the underlying driving force was the samefor each damage mode, with crack propagation being driven by an elastic-plastic mismatch between the metalmatrix and ceramic lamellae