The intracellular loop between domains I and II of the B type calcium channel confers aspects of G protein sensitivity to the E type calcium channel

Neuronal voltage-dependent calcium channels undergo inhibitory modulation by G-protein activation, generally involving both kinetic slowing and steady-state inhibition. We have shown previously that the b-subunit of neuronal calcium channels plays an important role in this process, because when it is absent, greater receptor-mediated inhibition is observed (Campbell et al., 1995b). We therefore hypothesized that the calcium channel b-subunits normally may occlude G-proteinmediated inhibition. Calcium channel b-subunits bind to the cytoplasmic loop between transmembrane domains I and II of the a1-subunits (Pragnell et al., 1994). We have examined the hypothesis that this loop is involved in G-protein-mediated inhibition by making chimeras containing the I–II loop of a1B or a1A inserted into a1E (a1EBE and a1EAE, respectively). This strategy was adopted because a1B (the molecular counterpart of N-type channels) and, to a lesser extent, a1A (P/Q-type) are G-protein-modulated, whereas this has not been observed to any great extent for a1E. Although a1B, coexpressed with a2-d and b1b transiently expressed in COS-7 cells, showed both kinetic slowing and steady-state inhibition when recorded with GTPgS in the patch pipette, both of which were reversed with a depolarizing prepulse, the chimera a1EBE (and, to a smaller extent, a1EAE) showed only kinetic slowing in the presence of GTPgS, and this also was reversed by a depolarizing prepulse. These results indicate that the I–II loop may be the molecular substrate of kinetic slowing but that the steady-state inhibition shown by a1B may involve a separate site on this calcium channel

Spatially and genetically distinct African trypanosome virulence variants defined by host interferon-g response

We describe 2 spatially distinct foci of human African trypansomiasis in eastern Uganda. The Tororo and Soroti foci of <i>Trypanosoma brucei rhodesiense</i> infection were genetically distinct as characterized by 6 microsatellite and 1 minisatellite polymorphic markers and were characterized by differences in disease progression and host-immune response. In particular, infections with the Tororo genotype exhibited an increased frequency of progression to and severity of the meningoencephalitic stage and higher plasma interferon (IFN)–γ concentration, compared with those with the Soroti genotype. We propose that the magnitude of the systemic IFN-γ response determines the time at which infected individuals develop central nervous system infection and that this is consistent with the recently described role of IFN-γ in facilitating blood-brain barrier transmigration of trypanosomes in an experimental model of infection. The identification of trypanosome isolates with differing disease progression phenotypes provides the first field-based genetic evidence for virulence variants in T. <i>brucei rhodesiense</i>

Determinants of the voltage dependence of G protein modulation within calcium channel β subunits

CaVβ subunits of voltage-gated calcium channels contain two conserved domains, a src-homology-3 (SH3) domain and a guanylate kinase-like (GK) domain with an intervening HOOK domain. We have shown in a previous study that, although Gβγ-mediated inhibitory modulation of CaV2.2 channels did not require the interaction of a CaVβ subunit with the CaVα1 subunit, when such interaction was prevented by a mutation in the α1 subunit, G protein modulation could not be removed by a large depolarization and showed voltage-independent properties (Leroy et al., J Neurosci 25:6984–6996, 2005). In this study, we have investigated the ability of mutant and truncated CaVβ subunits to support voltage-dependent G protein modulation in order to determine the minimal domain of the CaVβ subunit that is required for this process. We have coexpressed the CaVβ subunit constructs with CaV2.2 and α2δ-2, studied modulation by the activation of the dopamine D2 receptor, and also examined basal tonic modulation. Our main finding is that the CaVβ subunit GK domains, from either β1b or β2, are sufficient to restore voltage dependence to G protein modulation. We also found that the removal of the variable HOOK region from β2a promotes tonic voltage-dependent G protein modulation. We propose that the absence of the HOOK region enhances Gβγ binding affinity, leading to greater tonic modulation by basal levels of Gβγ. This tonic modulation requires the presence of an SH3 domain, as tonic modulation is not supported by any of the CaVβ subunit GK domains alone

Phosphorylation of the androgen receptor is associated with reduced survival in hormonerefractory prostate cancer patients

Cell line studies demonstrate that the PI3K/Akt pathway is upregulated in hormone-refractory prostate cancer (HRPC) and can result in phosphorylation of the androgen receptor (AR). The current study therefore aims to establish if this has relevance to the development of clinical HRPC. Immunohistochemistry was employed to investigate the expression and phosphorylation status of Akt and AR in matched hormone-sensitive and -refractory prostate cancer tumours from 68 patients. In the hormone-refractory tissue, only phosphorylated AR (pAR) was associated with shorter time to death from relapse (<i>P</i>=0.003). However, when an increase in expression in the transition from hormone-sensitive to -refractory prostate cancer was investigated, an increase in expression of PI3K was associated with decreased time to biochemical relapse (<i>P</i>=0.014), and an increase in expression of pAkt<sup>473</sup> and pAR<sup>210</sup> were associated with decreased disease-specific survival (<i>P</i>=0.0019 and 0.0015, respectively). Protein expression of pAkt<sup>473</sup> and pAR<sup>210</sup> also strongly correlated (<i>P</i><0.001, c.c.=0.711) in the hormone-refractory prostate tumours. These results provide evidence using clinical specimens, that upregulation of the PI3K/Akt pathway is associated with phosphorylation of the AR during development of HRPC, suggesting that this pathway could be a potential therapeutic target

The Herschel Exploitation of Local Galaxy Andromeda (HELGA) II: Dust and Gas in Andromeda

We present an analysis of the dust and gas in Andromeda, using Herschel images sampling the entire far-infrared peak. We fit a modified-blackbody model to ~4000 quasi-independent pixels with spatial resolution of ~140pc and find that a variable dust-emissivity index (beta) is required to fit the data. We find no significant long-wavelength excess above this model suggesting there is no cold dust component. We show that the gas-to-dust ratio varies radially, increasing from ~20 in the center to ~70 in the star-forming ring at 10kpc, consistent with the metallicity gradient. In the 10kpc ring the average beta is ~1.9, in good agreement with values determined for the Milky Way (MW). However, in contrast to the MW, we find significant radial variations in beta, which increases from 1.9 at 10kpc to ~2.5 at a radius of 3.1kpc and then decreases to 1.7 in the center. The dust temperature is fairly constant in the 10kpc ring (ranging from 17-20K), but increases strongly in the bulge to ~30K. Within 3.1kpc we find the dust temperature is highly correlated with the 3.6 micron flux, suggesting the general stellar population in the bulge is the dominant source of dust heating there. At larger radii, there is a weak correlation between the star formation rate and dust temperature. We find no evidence for 'dark gas' in M31 in contrast to recent results for the MW. Finally, we obtained an estimate of the CO X-factor by minimising the dispersion in the gas-to-dust ratio, obtaining a value of (1.9+/-0.4)x10^20 cm^-2 [K kms^-1]^-1.Comment: 19 pages, 18 figures. Submitted to ApJ April 2012; Accepted July 201

Bacterial microevolution and the Pangenome

The comparison of multiple genome sequences sampled from a bacterial population reveals considerable diversity in both the core and the accessory parts of the pangenome. This diversity can be analysed in terms of microevolutionary events that took place since the genomes shared a common ancestor, especially deletion, duplication, and recombination. We review the basic modelling ingredients used implicitly or explicitly when performing such a pangenome analysis. In particular, we describe a basic neutral phylogenetic framework of bacterial pangenome microevolution, which is not incompatible with evaluating the role of natural selection. We survey the different ways in which pangenome data is summarised in order to be included in microevolutionary models, as well as the main methodological approaches that have been proposed to reconstruct pangenome microevolutionary history

Quantitative Changes in Hydrocarbons over Time in Fecal Pellets of Incisitermes minor May Predict Whether Colonies Are Alive or Dead

Hydrocarbon mixtures extracted from fecal pellets of drywood termites are species-specific and can be characterized to identify the termites responsible for damage, even when termites are no longer present or are unable to be recovered easily. In structures infested by drywood termites, it is common to find fecal pellets, but difficult to sample termites from the wood. When fecal pellets appear after remedial treatment of a structure, it is difficult to determine whether this indicates that termites in the structure are still alive and active or not. We examined the hydrocarbon composition of workers, alates, and soldiers of Incisitermes minor (Hagen) (family Kalotermitidae) and of fecal pellets of workers. Hydrocarbons were qualitatively similar among castes and pellets. Fecal pellets that were aged for periods of 0, 30, 90, and 365 days after collection were qualitatively similar across all time periods, however, the relative quantities of certain individual hydrocarbons changed over time, with 19 of the 73 hydrocarbon peaks relatively increasing or decreasing. When the sums of the positive and negative slopes of these 19 hydrocarbons were indexed, they produced a highly significant linear correlation (R2 = 0.89). Consequently, the quantitative differences of these hydrocarbons peaks can be used to determine the age of worker fecal pellets, and thus help determine whether the colony that produced them is alive or dead

Reassuring and managing patients with concerns about swine flu: Qualitative interviews with callers to NHS Direct

<p>Abstract</p> <p>Background</p> <p>During the early stages of the 2009 swine flu (influenza H1N1) outbreak, the large majority of patients who contacted the health services about the illness did not have it. In the UK, the NHS Direct telephone service was used by many of these patients. We used qualitative interviews to identify the main reasons why people approached NHS Direct with concerns about swine flu and to identify aspects of their contact which were reassuring, using a framework approach.</p> <p>Methods</p> <p>33 patients participated in semi-structured interviews. All patients had telephoned NHS Direct between 11 and 14 May with concerns about swine flu and had been assessed as being unlikely to have the illness.</p> <p>Results</p> <p>Reasons for seeking advice about swine flu included: the presence of unexpectedly severe flu-like symptoms; uncertainties about how one can catch swine flu; concern about giving it to others; pressure from friends or employers; and seeking 'peace of mind.' Most participants found speaking to NHS Direct reassuring or useful. Helpful aspects included: having swine flu ruled out; receiving an alternative explanation for symptoms; clarification on how swine flu is transmitted; and the perceived credibility of NHS Direct. No-one reported anything that had increased their anxiety and only one participant subsequently sought additional advice about swine flu from elsewhere.</p> <p>Conclusions</p> <p>Future major incidents involving other forms of chemical, biological or radiological hazards may also cause large numbers of unexposed people to seek health advice. Our data suggest that providing telephone triage and information is helpful in such instances, particularly where advice can be given via a trusted, pre-existing service.</p

Thermodynamic instability of doubly spinning black objects

We investigate the thermodynamic stability of neutral black objects with (at least) two angular momenta. We use the quasilocal formalism to compute the grand canonical potential and show that the doubly spinning black ring is thermodynamically unstable. We consider the thermodynamic instabilities of ultra-spinning black objects and point out a subtle relation between the microcanonical and grand canonical ensembles. We also find the location of the black string/membrane phases of doubly spinning black objects.Comment: 25 pages, 7 figures v2: matches the published versio

Pan-Chromatic observations of the Recurrent Nova LMC 2009a (LMC 1971b)

Nova LMC 2009a is confirmed as a Recurrent Nova (RN) from positional coincidence with nova LMC 1971b. The observational data set is one of the most comprehensive for any Galactic or extragalactic RN: optical and near-IR photometry from outburst until over 6 years later; optical spectra for the first 6 months, and Swift satellite Ultraviolet and X-ray observations from 9 days to almost 1 year post-outburst. We find $M_V = -8.4\pm0.8_{\mathrm{r}}\pm0.7_{\mathrm{s}}$ and expansion velocities between 1000 and 4000 km s$^{-1}$. Coronal line emission before day 9 indicates shocks in the ejecta. Strengthening of He II $\lambda$4686 preceded the emergence of the Super-Soft Source (SSS) in X-rays at $\sim63-70$ days, which was initially very variable. Periodic modulations, $P=1.2$ days, most probably orbital in nature, were evident in the UV and optical from day 43. Subsequently, the SSS shows an oscillation with the same period but with a delay of 0.28P. The progenitor system has been identified; the secondary is most likely a sub-giant feeding a luminous accretion disk. Properties of the SSS infer a white dwarf (WD) mass $1.1 \mathrm{M}_\odot \lesssim M_{\rm WD} \lesssim 1.3 \mathrm{M}_\odot$. If the accretion occurs at constant rate, $\dot{\it{M}}_{\rm acc} \simeq 3.6^{+4.7}_{-2.5} \times 10^{-7} \mathrm{M}_\odot$ yr$^{-1}$ is needed, consistent with nova models for an inter-eruption interval of 38 years, low outburst amplitude, progenitor position in the color-magnitude diagram, and spectral energy distribution at quiescence. We note striking similarities between LMC 2009a and the Galactic nova KT Eri, suggesting that KT Eri is a candidate RN