Isolated pores dissected from human two-pore channel 2 are functional

Multi-domain voltage-gated ion channels appear to have evolved through sequential rounds of intragenic duplication from a primordial one-domain precursor. Whereas modularity within one-domain symmetrical channels is established, little is known about the roles of individual regions within more complex asymmetrical channels where the domains have undergone substantial divergence. Here we isolated and characterised both of the divergent pore regions from human TPC2, a two-domain channel that holds a key intermediate position in the evolution of voltage-gated ion channels. In HeLa cells, each pore localised to the ER and caused Ca2+ depletion, whereas an ER-targeted pore mutated at a residue that inactivates full-length TPC2 did not. Additionally, one of the pores expressed at high levels in E. coli. When purified, it formed a stable, folded tetramer. Liposomes reconstituted with the pore supported Ca2+ and Na+ uptake that was inhibited by known blockers of full-length channels. Computational modelling of the pore corroborated cationic permeability and drug interaction. Therefore, despite divergence, both pores are constitutively active in the absence of their partners and retain several properties of the wild-type pore. Such symmetrical ‘pore-only’ proteins derived from divergent channel domains may therefore provide tractable tools for probing the functional architecture of complex ion channels

Flamelet/flow interaction in premixed turbulent flames - Simultaneous measurements of gas velocity and flamelet position

An experimental technique for obtaining simultaneous measurements of fluid velocity and flamelet position in premixed flames is described and applied in a turbulent V-flame. The flamelet position information is used to calculate conditional velocity statistics, conditional on both zone (reactants or products) as well as conditional on distance from the flamelet. The conditional zone statistics demonstrate that increases (or decreases) in turbulence across the flame are dependent on axial position and location within the flame brush. The product- zone conditional covariance, coupled with the measured conditional mean velocity profiles, indicate that turbulence generation by shear may be a significant contribution to product zone turbulence levels. Velocity statistics conditional on distance from the flamelet demonstrate a considerable interaction between the flamelet and velocity field. Man and rms velocities vary significantly with proximity to the flamelet, such that differences in velocities which which occur just across the flamelet surface. The change in rms velocities just across the flamelet is found to be anisotropic, with the largest increase (smallest decrease) occurring in the axial velocity component. Rms velocities conditional on flamelet position further support the hypothesis that increased product gas velocity fluctuations may have a significant component associated with turbulence generation by mean shear

Time resolved amplified FRET identifies protein kinase B activation state as a marker for poor prognosis in clear cell renal cell carcinoma

Purpose Clear cell Renal Cell Carcinomas (ccRCC), the largest group of renal tumours, are resistant to classical therapies. The determination of the functional state of actionable biomarkers for the assessment of these adenocarcinomas is essential. The dysregulation of the oncoprotein, PKB/Akt has been linked with poor prognoses in human cancers. Material & methods We analysed the status of the PKB/Akt pathway in a representative tumour tissue microarray obtained from the primary tumours and their metastases in 60 ccRCC with long term follow up. We sought to define the evolution of this pathway from the primary tumour to the metastatic event and to know the impact of its functional state in tumour aggressiveness and patient survival. Two-site time resolved amplified FRET (A-FRET) was utilised for assessing the activation state of PKB/Akt and this was compared to conventional immunohistochemistry measurements. Results Activation state of PKB/Akt in primary tumours defined by A-FRET correlated with poorer overall survival (hazard ratio 0.228; p = 0.002). Whereas, increased protein expression of phosphoPKB/Akt, identified using classical immunohistochemistry, yielded no significant difference (hazard ratio 1.390; p = 0.548). Conclusions Quantitative determination of PKB/Akt activation in ccRCC primary tumours alongside other diagnostics tools could prove key in taking oncologists closer to an efficient personalised therapy in ccRCC patients. General significance The quantitative imaging technology based on Amplified-FRET can rapidly analyse protein activation states and molecular interactions. It could be used for prognosis and assess drug function during the early cycles of chemotherapy. It enables evaluation of clinical efficiency of personalised cancer treatment

Patient experiences of anxiety, depression and acute pain after surgery: a longitudinal perspective

This study sought to explore the impact of the psychological variables anxiety and depression, on pain experience over time following surgery. Eighty-five women having major gynaecological surgery were assessed for anxiety, depression and pain after surgery. To gain further understanding, 37 patients participated in a semi-structured taped telephone interview 4–6 weeks post-operatively. Pre-operative anxiety was found to be predictive of post-operative anxiety on Day 2, with patients who experienced high levels of anxiety before surgery continuing to feel anxious afterwards. By Day 4 both anxiety and depression scores increased as pain increased and one-third of the sample experienced levels of anxiety in psychiatric proportions whilst under one-third experienced similar levels of depression. These findings have significant implications for the provision of acute pain management after surgery. Future research and those managing acute pain services need to consider the multidimensional effect of acute pain and the interface between primary and secondary care

Resonantly damped surface and body MHD waves in a solar coronal slab with oblique propagation

The theory of magnetohydrodynamic (MHD) waves in solar coronal slabs in a zero-$\beta$ configuration and for parallel propagation of waves does not allow the existence of surface waves. When oblique propagation of perturbations is considered both surface and body waves are able to propagate. When the perpendicular wave number is larger than a certain value, the body kink mode becomes a surface wave. In addition, a sausage surface mode is found below the internal cut-off frequency. When non-uniformity in the equilibrium is included, surface and body modes are damped due to resonant absorption. In this paper, first, a normal-mode analysis is performed and the period, the damping rate, and the spatial structure of eigenfunctions are obtained. Then, the time-dependent problem is solved, and the conditions under which one or the other type of mode is excited are investigated.Comment: 19 pages, 9 figures, accepted for publication in Solar Physic

Tectonic controls on post-subduction granite genesis and emplacement : the late Caledonian suite of Britain and Ireland

Rates of magma emplacement commonly vary as a function of tectonic setting. The late Caledonian granites of Britain and Ireland are associated with closure of the Iapetus Ocean and were emplaced into a varying regime of transpression and transtension throughout the Silurian and into the early Devonian. Here we evaluate a new approach for examining how magma volumes vary as a function of tectonic setting. Available radiometric ages from the late Caledonian granites are used to calculate probability density functions (age spectra), with each pluton weighted by outcrop area as a proxy for its volume. These spectra confirm an absence of magmatic activity during Iapetus subduction between c. 455 Ma and 425 Ma and a dominance of post-subduction magmas between c. 425 Ma and 380 Ma. We review possible reasons why, despite the widespread outcrop of the late Caledonian granites, magmatism appears absent during Iapetus subduction. These include shallow angle subduction or extensive erosion and tectonic removal of the arc. In contrast to previous work we find no strong difference in the age or major element chemistry of post-subduction granites across all terranes. We propose a common causal mechanism in which the down-going Iapetus oceanic slab peeled back and detached beneath the suture following final Iapetus closure. The lithospheric mantle was delaminated beneath the suture and for about 100 km back beneath the Avalonian margin. While magma generation is largely a function of gravitationally driven lithosphere delamination, strike-slip dominated kinematics in the overlying continental crust is what modulated granitic magma emplacement. Early Devonian (419–404 Ma) transtension permitted large volumes of granite emplacement, whereas the subsequent Acadian (late Early Devonian, 404–394 Ma) transpression reduced and eventually suppressed magma emplacement

Tight-binding parameters for charge transfer along DNA

We systematically examine all the tight-binding parameters pertinent to charge transfer along DNA. The $\pi$ molecular structure of the four DNA bases (adenine, thymine, cytosine, and guanine) is investigated by using the linear combination of atomic orbitals method with a recently introduced parametrization. The HOMO and LUMO wavefunctions and energies of DNA bases are discussed and then used for calculating the corresponding wavefunctions of the two B-DNA base-pairs (adenine-thymine and guanine-cytosine). The obtained HOMO and LUMO energies of the bases are in good agreement with available experimental values. Our results are then used for estimating the complete set of charge transfer parameters between neighboring bases and also between successive base-pairs, considering all possible combinations between them, for both electrons and holes. The calculated microscopic quantities can be used in mesoscopic theoretical models of electron or hole transfer along the DNA double helix, as they provide the necessary parameters for a tight-binding phenomenological description based on the $\pi$ molecular overlap. We find that usually the hopping parameters for holes are higher in magnitude compared to the ones for electrons, which probably indicates that hole transport along DNA is more favorable than electron transport. Our findings are also compared with existing calculations from first principles.Comment: 15 pages, 3 figures, 7 table

Natural micropolymorphism in human leukocyte antigens provides a basis for genetic control of antigen recognition

Human leukocyte antigen (HLA) gene polymorphism plays a critical role in protective immunity, disease susceptibility, autoimmunity, and drug hypersensitivity, yet the basis of how HLA polymorphism influences T cell receptor (TCR) recognition is unclear. We examined how a natural micropolymorphism in HLA-B44, an important and large HLA allelic family, affected antigen recognition. T cell–mediated immunity to an Epstein-Barr virus determinant (EENLLDFVRF) is enhanced when HLA-B*4405 was the presenting allotype compared with HLA-B*4402 or HLA-B*4403, each of which differ by just one amino acid. The micropolymorphism in these HLA-B44 allotypes altered the mode of binding and dynamics of the bound viral epitope. The structure of the TCR–HLA-B*4405EENLLDFVRF complex revealed that peptide flexibility was a critical parameter in enabling preferential engagement with HLA-B*4405 in comparison to HLA-B*4402/03. Accordingly, major histocompatibility complex (MHC) polymorphism can alter the dynamics of the peptide-MHC landscape, resulting in fine-tuning of T cell responses between closely related allotypes

Anthropogenic Space Weather

Anthropogenic effects on the space environment started in the late 19th century and reached their peak in the 1960s when high-altitude nuclear explosions were carried out by the USA and the Soviet Union. These explosions created artificial radiation belts near Earth that resulted in major damages to several satellites. Another, unexpected impact of the high-altitude nuclear tests was the electromagnetic pulse (EMP) that can have devastating effects over a large geographic area (as large as the continental United States). Other anthropogenic impacts on the space environment include chemical release ex- periments, high-frequency wave heating of the ionosphere and the interaction of VLF waves with the radiation belts. This paper reviews the fundamental physical process behind these phenomena and discusses the observations of their impacts.Comment: 71 pages, 35 figure