Incremental prognostic value of the exercise electrocardiogram in the initial assessment of patients with suspected angina: cohort study

Objective To determine whether resting and exercise electrocardiograms (ECGs) provide prognostic value that is incremental to that obtained from the clinical history in ambulatory patients with suspected angina attending chest pain clinics

Characterising restrictions on commercial advertising and sponsorship of harmful commodities in local government policies: a nationwide study in England.

BackgroundCommercial advertising and sponsorship drive the consumption of harmful commodities. Local authorities (LAs) have considerable powers to reduce such exposures. This study aimed to characterize local commercial policies across all English LAs.MethodsWe conducted a census of all English LAs (n = 333) to identify local commercial policies concerning advertising and sponsorship of tobacco, alcohol, less healthy foods and gambling, through online searches and Freedom of Information requests. We explored policy presence, commodity frequency and type, and associations with LA characteristics (region, urban/rural and deprivation).ResultsOnly a third (106) of LAs in England had a relevant policy (32%). These included restrictions on tobacco (91%), gambling (79%), alcohol (74%) and/or less healthy foods (24%). Policy prevalence was lowest in the East of England (22%), North East (25%) and North West (27%), higher in urban areas (36%) than rural areas (28%) and lower in the least (27%) compared with the most (38%) deprived areas. Definitions in policies varied, particularly for alcohol and less healthy foods.ConclusionsEnglish LAs currently underutilize their levers to reduce the negative impacts of harmful commodity industry marketing, particularly concerning less healthy foods. Standardized guidance, including clarity on definitions and application, could inform local policy development

Cadmium interferes with auxin physiology and lignification in poplar

Cadmium (Cd) is a phytotoxic heavy metal that causes rapid growth reduction. To investigate if Cd interferes with the metabolism of auxin, a major growth hormone in plants, poplars (Populus×canescens) expressing a heterologous GH3::GUS reporter gene were exposed to 50 μM Cd in hydroponic solutions. Growth, photosynthetic performance, lignification, peroxidase activity, auxin concentration, and GUS staining were determined in order to record the activities of GH3 enzymes in the stem apex, the elongation zone, wood in the zone of radial growth, and in roots. Cd-induced growth reductions were tissue-specific decreasing in the order: roots>wood>shoot elongation and leaf initiation, whereas Cd concentrations increased in the order: leaves<wood<roots. Cd almost abolished the GH3 signal in the stem apex but caused strong increases in the vascular system of roots as well as in parenchymatic cells in the xylem. These changes were accompanied by increases in lignin and peroxidase activities and decreases in auxin concentrations. Since GH3 enzymes remove auxin from the active pool by conjugation and act as mediators between growth and defence, our data suggest that Cd stress triggered increases in GH3 activities which, in turn, depleted auxin in wood and thereby shunted the metabolism to enhanced formation of lignin

The 2015 Annual Meeting of SETAC German Language Branch in Zurich (7-10 September, 2015): ecotoxicology and environmental chemistry-from research to application

This report provides a brief review of the 20th annual meeting of the German Language Branch of the Society of Environmental Toxicology and Chemistry (SETAC GLB) held from September 7th to 10th 2015 at ETH (Swiss Technical University) in Zurich, Switzerland. The event was chaired by Inge Werner, Director of the Swiss Centre for Applied Ecotoxicology (Ecotox Centre) Eawag-EPFL, and organized by a team from Ecotox Centre, Eawag, Federal Office of the Environment, Federal Office of Agriculture, and Mesocosm GmbH (Germany). Over 200 delegates from academia, public agencies and private industry of Germany, Switzerland and Austria attended and discussed the current state of science and its application presented in 75 talks and 83 posters. In addition, three invited keynote speakers provided new insights into scientific knowledge ‘brokering’, and—as it was the International Year of Soil—the important role of healthy soil ecosystems. Awards were presented to young scientists for best oral and poster presentations, and for best 2014 master and doctoral theses. Program and abstracts of the meeting (mostly in German) are provided as Additional file 1

Untersuchungen zur Mobilität der Na,K-ATPase in Zellmembranen mittels Fluoreszenzmikroskopie

The Na,K-ATPase is a plasma membrane ion transporting enzyme, which is of high physiological importance especially in excitable cells of the central nervous system. It converts the energy from ATP hydrolysis to electrochemical gradients of Na+ and K+ ions across the cell membrane. The enzyme transports three sodium ions out of the cell and two potassium ions into the cell for each hydrolyzed ATP molecule. Mutations in genes coding for Na,K-ATPase isoforms lead to severe human pathologies like familial hemiplegic migraine (FHM), alternating hemiplegia of childhood (AHC) or epilepsy. Many of the reported mutations lead to loss-of-function effects on the molecular level, but others do not alter the function of the enzyme. Instead, to e.g. reduced protein stability, reduced protein expression or defective cellular localization may result. Therefore, the overall understanding of the implementation of the Na,K-ATPase into the plasma membrane is of great interest. By mutational disruption of binding sites of the Na,KATPase with ankyrin B (NaK-K456E) and caveolin-1 (NaK-ΔC, NaK-ΔN) it could be expected to observe changes in the plasma membrane targeting of the enzyme and changes of the localization within the plasma membrane and, consequently, of the diffusion behavior of the enzyme. The diffusion behavior of the different Na,K-ATPase constructs in the plasma membrane of living HEK293T cells was studied with fluorescence correlation spectroscopy (FCS) and as well with fluorescence recovery after photobleaching (FRAP). Both methods require labeling of the observed proteins with suitable fluorescent markers. Since from the literature it is known that common genetically encoded fluorescent proteins, e.g. eGFP, lead to artifacts in diffusion studies by FRAP due to photoswitching, the reversibly photoswitchable fluorescent protein Dreiklang (DRK) was used in addition as fluorescent marker. FCS studies on the Na,K-ATPase labeled with DRK lead to the conclusion that DRK is unsuitable for FCS analysis. It seems that DRK undergoes photophysical quenching processes, which occur on the same time scale as the observed diffusion times and, thus, small differences in diffusion are not measurable. The different Na,K-ATPase mutants labeled with eGFP exhibit significant differences in the diffusion behavior within the plasma membrane compared with the Na,K-ATPase wild-type. FRAP measurements on the Na,K-ATPase labeled with eGFP do not show significantly different results, but affirm the problems mentioned above. On the other side, DRK as fluorescence marker studied with standard FRAP settings also exhibits artificially shortened diffusion components, which can be attributed to reversible photoswitching of the fluorescent proteins and, therefore, these data are biased by photophysical artifacts. This problem is overcome by reversible FRAP experiments with DRK as fluorescence marker. However, these reversible FRAP data should be considered as a first qualitatively result and gives a hint about the different diffusion behavior of the various Na,K-ATPase constructs, because a thermically reactivation of the OFF-switched proteins into the ON-state is possible. These measurements open another view on the study of the implementation of the Na,KATPase into the plasma membrane and provides the opportunity to gain a deeper understanding of the binding or fixation of this enzyme within the membrane. FCS measurements can be therefore used in future investigations on Na,K-ATPase mutants linked to several diseases.Die Natrium-Kalium-ATPase (Na,K-ATPase) ist eine spannungsabhängige Ionenpumpe und eines der bedeutendsten Membrantransport-Proteine, welche insbesondere in erregbaren Zellen des zentralen Nervensystems von hoher physiologischer Bedeutung sind. Die Na,K-ATPase transportiert, gegen den chemischen Konzentrationsgradienten unter ATP Hydrolyse, drei Natriumionen aus der Zelle und zwei Kaliumionen in die Zelle. Diese beiden Prozesse sind strikt aneinander gekoppelt und halten somit die elektrochemischen Gradienten für Na+- und K+-Ionen über die Plasmamembran der Zelle aufrecht. Mutationen in den Genen der verschiedenen Isoformen der Na,K-ATPase α-Untereinheit können zu schweren Erkrankungen wie familiärer hemiplegischer Migräne (FHM Typ 2), alternating hemiplegia of childhood (AHC) oder Epilepsie führen. Viele der bereits bekannten Mutationen führen zum Verlust ihrer Funktion auf molekularer Ebene, wohingegen andere die Funktion des Enzyms nicht verändern oder beeinflussen. Stattdessen verursachen diese Mutationen eine reduzierte Proteinstabilität, verringern die Proteinexpression oder führen zur defekten zellulären Lokalisation. In dieser Arbeit wurden verschiedene Mutationen in der Na,K-ATPase α2-Untereinheit untersucht, die mit der Bindung der Na,K-ATPase an andere membranständige Proteine, wie z.B. Caveolin-1 (NaK-ΔC, NaK-ΔN), oder an Bindungsproteine des Zytoskeletts, wie Ankyrin B (NaK-K456E) assoziiert werden sowie der Fixierung innerhalb der Plasmamembran dienen. Es wird angenommen, dass diese Mutationen zu veränderter Zielsteuerung des Enzyms, Veränderung der Lokalisierung in der Plasmamembran und damit zur Änderung des Diffusionsverhaltens des Enzyms führen. Das Diffusionsverhalten der verschiedenen Konstrukte in der Plasmamembran von lebenden HEK293 Zellen wurde mit zwei verschiedenen Messtechniken untersucht, zum einen mit der Fluoreszenzkorrelationsspektroskopie (FCS) und zum anderen mit Fluorescence recovery after photobleaching (FRAP). Beide Verfahren erfordern die Markierung der zu beobachtenden Proteine mit geeigneten Fluoreszenzmarkern. Da in der Literatur beschrieben ist, dass genetisch kodierte Fluoreszenzproteine wie das grün fluoreszierende Protein (GFP) zu Artefakten durch Photoschalten in FRAP-Messungen führen, wurde außerdem das reversibel schaltbare Dreiklang Protein (DRK) als Marker verwendet. Es konnte gezeigt werden, dass DRK als Marker für FCS-Messungen nicht geeignet ist, da das Protein unbekannte photophysikalische Prozesse durchläuft, welche sich auf der gleichen Zeitskala, wie die zu erwartenden Diffusionsänderungen, abspielen und somit diese überlagern. Die verschiedenen eGFP-Na,K-ATPase Mutanten zeigten im Vergleich zum Wildtyp signifikante Unterschiede bei FCS-Messungen. Alle Mutanten wiesen eine deutlich schnellere Diffusion auf als der Wildtyp, was auf eine gelockerte Bindung an der Membran zurückgeführt werden kann. FRAP-Messungen unter Verwendung von eGFP als Fluoreszenzmarker zeigten deutlich eine sehr schnelle Diffusionskomponente, welche auf das reversible Photoschalten der Proteine zurückzuführen ist und damit die Messung für kleine Unterschiede im Diffusionsverhalten unmöglich machen. Die gleichen Ergebnisse wurden auch für DRK markierte Zellen unter Standard FRAP Bedingungen beobachtet. Dieses Problem konnte mit Hilfe von reversiblen FRAP-Messungen umgangen werden, indem DRK in seinen ausgeschalteten Zustand gebracht wurde, anstatt DRK irreversibel zu bleichen. Anhand dieser Messungen konnten signifikante Unterschiede der DRK-Na,K-ATPase Mutanten im Vergleich zum Wildtyp beobachtet werden. Es muss aber hier angemerkt werden, dass diese Messungen nur als qualitative Untersuchung angesehen werden können, da DRK durch thermische Reaktivierung wieder in den eingeschalteten Zustand zurückkehren kann und dieser Prozess die zu beobachtenden Diffusionsprozesse überlagert. Die in dieser Arbeit durchgeführten Messungen eröffnen eine neue Sichtweise für die Untersuchung der Na,K-ATPase und lassen Rückschlüsse auf die Fixierung innerhalb der Plasmamembran und die Bindung an andere Adapter- oder Matrixproteine zu. Dadurch kann in zukünftigen Untersuchungen die FCS Messtechnik genutzt werden um unbekannte Mutationen zu studieren, welche mit verschiedenen Krankheiten assoziiert werden und somit einen wertvollen Beitrag zur Erforschung deren Ursachen leisten können

Risk assessment after acute coronary syndrome

Lots of potential but will it end up being yet another risk score

Empowering community health professionals for effective air pollution information communication

Abstract Background Air pollution remains a significant public health risk, particularly in urban areas. Effective communication strategies remain integral to overall protection by encouraging the adoption of personal air pollution exposure reduction behaviours. This study aims to explore how community health professionals can be empowered to communicate air pollution information and advice to the wider community, to encourage the uptake of desired behaviours in the population. Methods The study adopted a qualitative methodology, where four homogenous Focus Group Discussions (FGDs) were held with a range of community health professionals, including Health Care Professionals, Community Health and Wellbeing Workers (CHWWs) and Social Prescribing Link Workers (SPLW). A classical content analysis was conducted with the Structural Empowerment Theory (SET) and Psychological Empowerment Theory (PET) as guiding concepts. Results Five key themes were identified: from a structural empowerment perspective: [1] resources and support, [2] knowledge. From a psychological empowerment perspective: [3] confidence as advisor, [4] responsibility as advisor, and [5] residents’ receptiveness to advice. It was concluded that advice should be risk stratified, clear, easy to follow and provide alternatives. Conclusion This study identified ways for community health professionals to be empowered by local councils or other organisations in providing advice on air pollution, through the provision of essential structural support and opportunities to enhance their knowledge and confidence in the subject. Implementing recommendations from this study would not only empower community health professionals to advise on air pollution to the wider community but also increase adherence to health advice

ATP1A2 Mutations in Migraine: Seeing through the Facets of an Ion Pump onto the Neurobiology of Disease

Mutations in four genes have been identified in familial hemiplegic migraine (FHM), from which CACNA1A (FHM type 1) and SCN1A (FHM type 3) code for neuronal voltage-gated calcium or sodium channels, respectively, while ATP1A2 (FHM type 2) encodes the α2 isoform of the Na+,K+-ATPase's catalytic subunit, thus classifying FHM primarily as an ion channel/ion transporter pathology. FHM type 4 is attributed to mutations in the PRRT2 gene, which encodes a proline-rich transmembrane protein of as yet unknown function. The Na+,K+-ATPase maintains the physiological gradients for Na+ and K+ ions and is, therefore, critical for the activity of ion channels and transporters involved neuronal excitability, neurotransmitter uptake or Ca2+ signaling. Strikingly diverse functional abnormalities have been identified for disease-linked ATP1A2 mutations which frequently lead to changes in the enzyme's voltage-dependent properties, kinetics, or apparent cation affinities, but some mutations are truly deleterious for enzyme function and thus cause full haploinsufficiency. Here, we summarize structural and functional data about the Na+,K+-ATPase available to date and an overview is provided about the particular properties of the α2 isoform that explain its physiological relevance in electrically excitable tissues. In addition, current concepts about the neurobiology of migraine, the correlations between primary brain dysfunction and mechanisms of headache pain generation are described, together with insights gained recently from modeling approaches in computational neuroscience. Then, a survey is given about ATP1A2 mutations implicated in migraine cases as documented in the literature with focus on mutations that were described to completely destroy enzyme function, or lead to misfolded or mistargeted protein in particular model cell lines. We also discuss whether or not there are correlations between these most severe mutational effects and clinical phenotypes. Finally, perspectives for future research on the implications of Na+,K+-ATPase mutations in human pathologies are presented.DFG, 53182490, EXC 314: Unifying Concepts in Catalysi