Delineating Zinc Influx Mechanisms during Platelet Activation

Zinc (Zn2+) is released by platelets during a hemostatic response to injury. Extracellular zinc ([Zn2+]o) initiates platelet activation following influx into the platelet cytosol. However, the mechanisms that permit Zn2+ influx are unknown. Fluctuations in intracellular zinc ([Zn2+]i) were measured in fluozin-3-loaded platelets using fluorometry and flow cytometry. Platelet activation was assessed using light transmission aggregometry. The detection of phosphoproteins was performed by Western blotting. [Zn2+]o influx and subsequent platelet activation were abrogated by blocking the sodium/calcium exchanged, TRP channels, and ZIP7. Cation store depletion regulated Zn2+ influx. [Zn2+]o stimulation resulted in the phosphorylation of PKC substates, MLC, and β3 integrin. Platelet activation via GPVI or Zn2+ resulted in ZIP7 phosphorylation in a casein kinase 2-dependent manner and initiated elevations of [Zn2+]i that were sensitive to the inhibition of Orai1, ZIP7, or IP3R-mediated pathways. These data indicate that platelets detect and respond to changes in [Zn2+]o via influx into the cytosol through TRP channels and the NCX exchanger. Platelet activation results in the externalization of ZIP7, which further regulates Zn2+ influx. Increases in [Zn2+]i contribute to the activation of cation-dependent enzymes. Sensitivity of Zn2+ influx to thapsigargin indicates a store-operated pathway that we term store-operated Zn2+ entry (SOZE). These mechanisms may affect platelet behavior during thrombosis and hemostasis

The role of plasma membrane STIM1 and Ca(2+)entry in platelet aggregation. STIM1 binds to novel proteins in human platelets.

Ca(2+) elevation is essential to platelet activation. STIM1 senses Ca(2+) in the endoplasmic reticulum and activates Orai channels allowing store-operated Ca(2+) entry (SOCE). STIM1 has also been reported to be present in the plasma membrane (PM) with its N-terminal region exposed to the outside medium but its role is not fully understood. We have examined the effects of the antibody GOK/STIM1, which recognises the N-terminal region of STIM1, on SOCE, agonist-stimulated Ca(2+) entry, surface exposure, in vitro thrombus formation and aggregation in human platelets. We also determined novel binding partners of STIM1 using proteomics. The dialysed GOK/STIM1 antibody failed to reduced thapsigargin- and agonist-mediated Ca(2+) entry in Fura2-labelled cells. Using flow cytometry we detect a portion of STIM1 to be surface-exposed. The dialysed GOK/STIM1 antibody reduced thrombus formation by whole blood on collagen-coated capillaries under flow and platelet aggregation induced by collagen. In immunoprecipitation experiments followed by proteomic analysis, STIM1 was found to extract a number of proteins including myosin, DOCK10, thrombospondin-1 and actin. These studies suggest that PM STIM1 may facilitate platelet activation by collagen through novel interactions at the plasma membrane while the essential Ca(2+)-sensing role of STIM1 is served by the protein in the ER

Empirical validation of the InVEST water yield ecosystem service model at a national scale

A variety of tools have emerged with the goal of mapping the current delivery of ecosystem services and quantifying the impact of environmental changes. An important and often overlooked question is how accurate the outputs of these models are in relation to empirical observations. In this paper we validate a hydrological ecosystem service model (InVEST Water Yield Model) using widely available data. We modelled annual water yield in 22 UK catchments with widely varying land cover, population and geology, and compared model outputs with gauged river flow data from the UK National River Flow Archive. Values for input parameters were selected from existing literature to reflect conditions in the UK and were subjected to sensitivity analyses. We also compared model performance between precipitation and potential evapotranspiration data sourced from global- and UK-scale datasets. We then tested the transferability of the results within the UK by additional validation in a further 20 catchments. Whilst the model performed only moderately with global-scale data (linear regression of modelled total water yield against empirical data; slope = 0.763, intercept = 54.45, R2 = 0.963) with wide variation in performance between catchments, the model performed much better when using UK-scale input data, with closer fit to the observed data (slope = 1.07, intercept = 3.07, R2 = 0.990). With UK data the majority of catchments showed less than 10% difference between measured and modelled water yield but there was a minor but consistent overestimate per hectare (86 m3/ha/year). Additional validation on a further 20 UK catchments was similarly robust, indicating that these results are transferable within the UK. These results suggest that relatively simple models can give accurate measures of ecosystem services. However, the choice of input data is critical and there is a need for further validation in other parts of the worl

A study of the link between cosmic rays and clouds with a cloud chamber at the CERN PS

Recent satellite data have revealed a surprising correlation between galactic cosmic ray (GCR) intensity and the fraction of the Earth covered by clouds. If this correlation were to be established by a causal mechanism, it could provide a crucial step in understanding the long-sought mechanism connecting solar and climate variability. The Earth's climate seems to be remarkably sensitive to solar activity, but variations of the Sun's electromagnetic radiation appear to be too small to account for the observed climate variability. However, since the GCR intensity is strongly modulated by the solar wind, a GCR-cloud link may provide a sufficient amplifying mechanism. Moreover if this connection were to be confirmed, it could have profound consequences for our understanding of the solar contributions to the current global warming. The CLOUD (Cosmics Leaving OUtdoor Droplets) project proposes to test experimentally the existence a link between cosmic rays and cloud formation, and to understand the microphysical mechanism. CLOUD plans to perform detailed laboratory measurements in a particle beam at CERN, where all the parameters can be precisely controlled and measured. The beam will pass through an expansion cloud chamber and a reactor chamber where the atmosphere is to be duplicated by moist air charged with selected aerosols and trace condensable vapours. An array of external detectors and mass spectrometers is used to analyse the physical and chemical characteristics of the aerosols and trace gases during beam exposure. Where beam effects are found, the experiment will seek to evaluate their significance in the atmosphere by incorporating them into aerosol and cloud models.Recent satellite data have revealed a surprising correlation between galactic cosmic ray (GCR) intensity and the fraction of the Earth covered by clouds. If this correlation were to be established by a causal mechanism, it could provide a crucial step in understanding the long-sought mechanism connecting solar and climate variability. The Earth's climate seems to be remarkably sensitive to solar activity, but variations of the Sun's electromagnetic radiation appear to be too small to account for the observed climate variability. However, since the GCR intensity is strongly modulated by the solar wind, a GCR-cloud link may provide a sufficient amplifying mechanism. Moreover if this connection were to be confirmed, it could have profound consequences for our understanding of the solar contributions to the current global warming. The CLOUD (Cosmics Leaving OUtdoor Droplets) project proposes to test experimentally the existence a link between cosmic rays and cloud formation, and to understand the microphysical mechanism. CLOUD plans to perform detailed laboratory measurements in a particle beam at CERN, where all the parameters can be precisely controlled and measured. The beam will pass through an expansion cloud chamber and a reactor chamber where the atmosphere is to be duplicated by moist air charged with selected aerosols and trace condensable vapours. An array of external detectors and mass spectrometers is used to analyse the physical and chemical characteristics of the aerosols and trace gases during beam exposure. Where beam effects are found, the experiment will seek to evaluate their significance in the atmosphere by incorporating them into aerosol and cloud models

Fuel flexible power stations: Utilisation of ash co-products as additives for NOx emissions control

This work investigated the effects of different ash co-products on the combustion of solid fuels, in particular the fuel-nitrogen behaviour: The fuel-ash additive combinations investigated were: Firstly, biomass ashes added to bituminous coals, representative of those used in power stations; Secondly, a low reactivity coal; Thirdly, a high-N biomass (olive cake) was chosen as a high reactivity fuel and studied with a power-station pulverised coal fly ash as an additive. These five solid fuels have a wide fuel ratio FR (i.e. the ratio of fixed carbon to volatile matter content). The ash additives were a pulverised fly ash (PFA) and a furnace bottom ash (FBA) from wood pellet combustion in a UK power station. Fuels (with and without additives) were studied for nitrogen partitioning during (i) devolatilisation and for (ii) NOX formation during combustion, using two different electrically heated drop tube furnaces (DTF) operating at 1373 K. Devolatilisation was also studied via ballistic-heated thermogravimetric analysis (TGA). The extent of impact of additives on volatile yield under devolatilisation conditions was dependent on fuel ratio, high FR has the greatest increase in volatile release when co-feeding the additive. Under devolatilisation conditions, there is a correlation between volatile nitrogen and carbon conversion for all the fuels tested. Thus, additives liberate more volatile-nitrogen from the coals and also deliver enhanced carbon conversion. A mechanism is proposed whereby ultra-fine particles and vapours of reactive compounds from the additives interact with the reacting fuel/char particle and influence N-release during both devolatilisation and char burn-out. The enhanced conversion of fuel-nitrogen to volatile-nitrogen and the reduction of char-nitrogen can lead to reductions of NOX emissions in emissions-controlled furnaces. This approach could assist fuel-flexible power stations in achieving their NOX emission targets

Modeling thrombosis in silico: Frontiers, challenges, unresolved problems and milestones

Hemostasis is a complex physiological mechanism that functions to maintain vascular integrity under any conditions. Its primary components are blood platelets and a coagulation network that interact to form the hemostatic plug, a combination of cell aggregate and gelatinous fibrin clot that stops bleeding upon vascular injury. Disorders of hemostasis result in bleeding or thrombosis, and are the major immediate cause of mortality and morbidity in the world. Regulation of hemostasis and thrombosis is immensely complex, as it depends on blood cell adhesion and mechanics, hydrodynamics and mass transport of various species, huge signal transduction networks in platelets, as well as spatiotemporal regulation of the blood coagulation network. Mathematical and computational modeling has been increasingly used to gain insight into this complexity over the last 30 years, but the limitations of the existing models remain profound. Here we review state-of-the-art-methods for computational modeling of thrombosis with the specific focus on the analysis of unresolved challenges. They include: a) fundamental issues related to physics of platelet aggregates and fibrin gels; b) computational challenges and limitations for solution of the models that combine cell adhesion, hydrodynamics and chemistry; c) biological mysteries and unknown parameters of processes; d) biophysical complexities of the spatiotemporal networks' regulation. Both relatively classical approaches and innovative computational techniques for their solution are considered; the subjects discussed with relation to thrombosis modeling include coarse-graining, continuum versus particle-based modeling, multiscale models, hybrid models, parameter estimation and others. Fundamental understanding gained from theoretical models are highlighted and a description of future prospects in the field and the nearest possible aims are given. © 2018 Elsevier B.V

'Lambing ears': a blistering disorder affecting farmers at lambing time.

At lambing time some farmers experience blistering and crusting of the pinnae. This occupational disease, termed 'lambing ears', does not feature in the medical literature. Objectives To define the condition and explore its pathogenesis. Methods We obtained five biopsies from affected individuals and sent questionnaires to 69 farmers in the U.K. Farming communities abroad were also contacted. Results The eruption lasts for the duration of the lambing practice. The histological features are dominated by a pandermal perivascular and diffuse, predominantly T-cell lymphocytic infiltrate. Only the pinnae are affected and its incidence is related to the degree of involvement a farmer has with the animals around parturition. The condition also occurs, but less frequently, in farmers who are calving. Conclusions This occupational disease occurs with close contact to lambing ewes or calving cows. The histology and distribution are comparable with the juvenile spring eruption variant of polymorphic light eruption, but its demographics are unique

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