A P2X receptor from the tardigrade species Hypsibius dujardini with fast kinetics and sensitivity to zinc and copper

<p>Abstract</p> <p>Background</p> <p>Orthologs of the vertebrate ATP gated P2X channels have been identified in <it>Dictyostelium </it>and green algae, demonstrating that the emergence of ionotropic purinergic signalling was an early event in eukaryotic evolution. However, the genomes of a number of animals including <it>Drosophila melanogaster </it>and <it>Caenorhabditis elegans</it>, both members of the Ecdysozoa superphylum, lack P2X-like proteins, whilst other species such as the flatworm <it>Schistosoma mansoni </it>have P2X proteins making it unclear as to what stages in evolution P2X receptors were lost. Here we describe the functional characterisation of a P2X receptor (<it>Hd</it>P2X) from the tardigrade <it>Hypsibius dujardini </it>demonstrating that purinergic signalling is preserved in some ecdysozoa.</p> <p>Results</p> <p>ATP (EC₅₀~44.5 μM) evoked transient inward currents in <it>Hd</it>P2X with millisecond rates of activation and desensitisation. <it>Hd</it>P2X is antagonised by pyridoxal-phosphate-6-azophenyl-2',4' disulfonic acid (IC₅₀15.0 μM) and suramin (IC₅₀22.6 μM) and zinc and copper inhibit ATP-evoked currents with IC₅₀values of 62.8 μM and 19.9 μM respectively. Site-directed mutagenesis showed that unlike vertebrate P2X receptors, extracellular histidines do not play a major role in coordinating metal binding in <it>Hd</it>P2X. However, H306 was identified as playing a minor role in the actions of copper but not zinc. Ivermectin potentiated responses to ATP with no effect on the rates of current activation or decay.</p> <p>Conclusion</p> <p>The presence of a P2X receptor in a tardigrade species suggests that both nematodes and arthropods lost their P2X genes independently, as both traditional and molecular phylogenies place the divergence between Nematoda and Arthropoda before their divergence from Tardigrada. The phylogenetic analysis performed in our study also clearly demonstrates that the emergence of the family of seven P2X channels in human and other mammalian species was a relatively recent evolutionary event that occurred subsequent to the split between vertebrates and invertebrates. Furthermore, several characteristics of <it>Hd</it>P2X including fast kinetics with low ATP sensitivity, potentiation by ivermectin in a channel with fast kinetics and distinct copper and zinc binding sites not dependent on histidines make <it>Hd</it>P2X a useful model for comparative structure-function studies allowing a better understanding of P2X receptors in higher organisms.</p

Device grade microcrystalline silicon owing to reduced oxygen contamination

As-deposited undoped microcrystalline silicon (µc-Si:H) has in general a pronounced n-type behavior. Such a material is therefore often not appropriate for use in devices, such as p-i-n diodes, as an active, absorbing i layer or as channel material for thin-film transistors. In recent work, on p-i-n solar cells, this disturbing n-type character had been successfully compensated by the ``microdoping'' technique. In the present letter, it is shown that this n-type behavior is mainly linked to oxygen impurities; therefore, one can replace the technologically delicate microdoping technique by a purification method, that is much easier to handle. This results in a reduction of oxygen impurities by two orders of magnitude; it has, furthermore a pronounced impact on the electrical properties of µc-Si:H films and on device performance, as well. Additionally, these results prove that the unwanted donor-like states within µc-Si:H are mainly due to extrinsic impurities and not to structural native defects

Morphology of supported polymer electrolyte ultra-thin films: a numerical study

Morphology of polymer electrolytes membranes (PEM), e.g., Nafion, inside PEM fuel cell catalyst layers has significant impact on the electrochemical activity and transport phenomena that determine cell performance. In those regions, Nafion can be found as an ultra-thin film, coating the catalyst and the catalyst support surfaces. The impact of the hydrophilic/hydrophobic character of these surfaces on the structural formation of the films has not been sufficiently explored yet. Here, we report about Molecular Dynamics simulation investigation of the substrate effects on the ionomer ultra-thin film morphology at different hydration levels. We use a mean-field-like model we introduced in previous publications for the interaction of the hydrated Nafion ionomer with a substrate, characterized by a tunable degree of hydrophilicity. We show that the affinity of the substrate with water plays a crucial role in the molecular rearrangement of the ionomer film, resulting in completely different morphologies. Detailed structural description in different regions of the film shows evidences of strongly heterogeneous behavior. A qualitative discussion of the implications of our observations on the PEMFC catalyst layer performance is finally proposed

Enhancement of Sm3+emission by SnO2nanocrystals in the silica matrix

Silica xerogels containing Sm3+ions and SnO2nanocrystals were prepared in a sol–gel process. The image of transmission electron microscopy (TEM) shows that the SnO2nanocrystals are dispersed in the silica matrix. The X-ray diffraction (XRD) of the sample confirms the tetragonal phase of SnO2. The xerogels containing SnO2nanocrystals and Sm3+ions display the characteristic emission of Sm3+ions (4G5/2 → 6HJ(J = 5/2, 7/2, 9/2)) at the excitation of 335 nm which energy corresponds to the energy gap of the SnO2nanocrystals, while no emission of Sm3+ions can be observed for the samples containing Sm3+ions. The enhancement of the Sm3+emission is probably due to the energy transfer from SnO2nanocrystals to Sm3+ions

Machine learning classifies predictive kinematic features in a mouse model of neurodegeneration

Motor deficits are observed in Alzheimer’s disease (AD) prior to the appearance of cognitive symptoms. To investigate the role of amyloid proteins in gait disturbances, we characterized locomotion in APP-overexpressing transgenic J20 mice. We used three-dimensional motion capture to characterize quadrupedal locomotion on a treadmill in J20 and wild-type mice. Sixteen J20 mice and fifteen wild-type mice were studied at two ages (4- and 13-month). A random forest (RF) classification algorithm discriminated between the genotypes within each age group using a leave-one-out cross-validation. The balanced accuracy of the RF classification was 92.3 ± 5.2% and 93.3 ± 4.5% as well as False Negative Rate (FNR) of 0.0 ± 0.0% and 0.0 ± 0.0% for the 4-month and 13-month groups, respectively. Feature ranking algorithms identified kinematic features that when considered simultaneously, achieved high genotype classification accuracy. The identified features demonstrated an age-specific kinematic profile of the impact of APP-overexpression. Trunk tilt and unstable hip movement patterns were important in classifying the 4-month J20 mice, whereas patterns of shoulder and iliac crest movement were critical for classifying 13-month J20 mice. Examining multiple kinematic features of gait simultaneously could also be developed to classify motor disorders in humans

PEG Branched Polymer for Functionalization of Nanomaterials with Ultralong Blood Circulation

Nanomaterials have been actively pursued for biological and medical applications in recent years. Here, we report the synthesis of several new poly(ethylene glycol) grafted branched-polymers for functionalization of various nanomaterials including carbon nanotubes, gold nanoparticles (NP) and gold nanorods (NRs), affording high aqueous solubility and stability for these materials. We synthesize different surfactant polymers based upon poly-(g-glutamic acid) (gPGA) and poly(maleic anhydride-alt-1-octadecene) (PMHC18). We use the abundant free carboxylic acid groups of gPGA for attaching lipophilic species such as pyrene or phospholipid, which bind to nanomaterials via robust physisorption. Additionally, the remaining carboxylic acids on gPGA or the amine-reactive anhydrides of PMHC18 are then PEGylated, providing extended hydrophilic groups, affording polymeric amphiphiles. We show that single-walled carbon nanotubes (SWNTs), Au NPs and NRs functionalized by the polymers exhibit high stability in aqueous solutions at different pHs, at elevated temperatures and in serum. Morever, the polymer-coated SWNTs exhibit remarkably long blood circulation (t1/2 22.1 h) upon intravenous injection into mice, far exceeding the previous record of 5.4 h. The ultra-long blood circulation time suggests greatly delayed clearance of nanomaterials by the reticuloendothelial system (RES) of mice, a highly desired property for in vivo applications of nanomaterials, including imaging and drug delivery