Transcriptome profiling reveals exposure to predicted end-of-century ocean acidification as a stealth stressor for Atlantic cod larvae

Ocean acidification (OA), a direct consequence of increasing atmospheric CO2 concentration dissolving in ocean waters, is impacting many fish species. Little is known about the molecular mechanisms underlying the observed physiological impacts in fish. We used RNAseq to characterize the transcriptome of 3 different larval stages of Atlantic cod (Gadus morhua) exposed to simulated OA at levels (1179 µatm CO2) representing end-of-century predictions compared to controls (503 µatm CO2), which were shown to induce tissue damage and elevated mortality in G. morhua. Only few genes were differentially expressed in 6 and 13 days-post-hatching (dph) (3 and 16 genes, respectively), during a period when maximal mortality as a response to elevated pCO2 occurred. At 36 dph, 1413 genes were differentially expressed, most likely caused by developmental asynchrony between the treatment groups, with individuals under OA growing faster. A target gene analysis revealed only few genes of the universal and well-defined cellular stress response to be differentially expressed. We thus suggest that predicted ocean acidification levels constitute a “stealth stress” for early Atlantic cod larvae, with a rapid breakdown of cellular homeostasis leading to organismal death that was missed even with an 8-fold replication implemented in this study

Impacts of highway traffic exhaust in alpine valleys on the respiratory health in adults: a cross-sectional study

BACKGROUND: Most studies having shown respiratory health effects from traffic exhaust were conducted in urban areas with a complex mixture of air pollution sources. This study has investigated the potential impact of traffic exhaust on respiratory symptoms among adults living along a Swiss alpine highway corridor, where traffic exhaust from the respective trans-Alpine highway is the predominant source of air pollution. METHODS: In summer 2005, we recruited 1839 adults aged 15 to 70 from a random sample of 10 communities along the Swiss alpine highway corridors. Subjects answered a questionnaire on respiratory health (asthmatic and bronchitic symptoms), risk factors, and potential confounding variables. We used logistic regression models to assess associations between respiratory symptoms and traffic exposure being defined a) as living within 200 m of the highway, and b) as a bell-shaped function simulating the decrease of pollution levels with increasing distance to the highway. RESULTS: Positive associations were found between living close to a highway and wheezing without cold (OR = 3.10, 95%-CI: 1.27-7.55) and chronic cough (OR = 2.88, 95%-CI: 1.17-7.05). The models using a bell-shaped function suggested that symptoms reached background levels after 400-500 m from the highway. The association with chronic cough was driven by a subgroup reporting hay fever or allergic rhinitis. CONCLUSIONS: Highway traffic exhaust in alpine highway corridors, in the absence of other industrial sources, showed negative associations with the respiratory health of adults, higher than those previously found in urban areas

Cerebrospinal fluid biomarker candidates associated with human WNV neuroinvasive disease

During the last decade, the epidemiology of WNV in humans has changed in the southern regions of Europe, with high incidence of West Nile fever (WNF) cases, but also of West Nile neuroinvasive disease (WNND). The lack of human vaccine or specific treatment against WNV infection imparts a pressing need to characterize indicators associated with neurological involvement. By its intimacy with central nervous system (CNS) structures, modifications in the cerebrospinal fluid (CSF) composition could accurately reflect CNS pathological process. Until now, few studies investigated the association between imbalance of CSF elements and severity of WNV infection. The aim of the present study was to apply the iTRAQ technology in order to identify the CSF proteins whose abundances are modified in patients with WNND. Forty-seven proteins were found modified in the CSF of WNND patients as compared to control groups, and most of them are reported for the first time in the context of WNND. On the basis of their known biological functions, several of these proteins were associated with inflammatory response. Among them, Defensin-1 alpha (DEFA1), a protein reported with anti-viral effects, presente

Hedgehog partial agonism drives warburg-lie metabolism in muscle and brown fat

Diabetes, obesity, and cancer affect upward of 15% of the world&rsquo;s population. Interestingly, all three diseases juxtapose dysregulated intracellular signaling with altered metabolic state. Exactly which genetic factors define stable metabolic set points in vivo remains poorly understood. Here, we show that hedgehog signaling rewires cellular metabolism. We identify a cilium-dependent Smo-Ca2+-Ampk axis that triggers rapid Warburg-like metabolic reprogramming within minutes of activation and is required for proper metabolic selectivity and flexibility. We show that Smo modulators can uncouple the Smo-Ampk axis from canonical signaling and identify cyclopamine as one of a new class of &ldquo;selective partial agonists,&rdquo; capable of concomitant inhibition of canonical and activation of noncanonical hedgehog signaling. Intriguingly, activation of the Smo-Ampk axis in vivo drives robust insulin-independent glucose uptake in muscle and brown adipose tissue. These data identify multiple noncanonical endpoints that are pivotal for rational design of hedgehog modulators and provide a new therapeutic avenue for obesity and diabetes.<br /

Quantitative (phospho)-proteomics identifies a novel role for hedgehog in reprogramming adipocyte energy metabolism

Zsfassung in dt. SpracheDer Hedgehog Signaltransduktionsweg (Hh) spielt eine essentielle Rolle in Entwicklungsprozessen von der Fruchtfliege bis hin zum Mensch.Eine unzureichende Funktion des Hh führt zu schweren Missbildungen, wobei eine Überreaktion mit Tumorentstehung in Verbindung steht.Daten unserer Forschungsarbeiten bestätigten (i) eine Schlüsselfunktion des Hh in der Inhibierung der Differenzierung weißer, aber nicht brauner Fettzellen sowohl in vitro als auch in vivo und, (ii) dass diese hemmende Wirkung durch das T-Zellen Zytokin Interferon-gamma blockiert wird. Interessanterweise hat sich bis dato keine Studie damit befasst, den Hh auf reife Fettzellen zu untersuchen und keine (phospho)-proteomischen Daten sind vorhanden. Somit setzten wir uns das Ziel, neue Mechanismen des Hh auf reife Fettzellen, mit Hilfe quantitativer proteomischer Methoden zu erfassen.2D-PAGE und 1D-GeLC/MSMS mit (i) angereicherten Phosphoproteinen und (ii) der zytoplasmatischen Fraktion reifer Adipozyten wurde angewandt.Ziel war es, Änderungen in (De)-Phosphorylierungen beziehungsweise des Expressionsmusters in Hedgehog stimulierten Fettzellen zu detektieren.Western blot Analysen, sowie ELISA Messungen untermauerten diese Ergebnisse.Phosphoprotein-spezifische 1D- und 2D-Gele identifizierten eine hoch reproduzierbare Phosphorylierung der Pyruvate Dehydrogenase (PDH) E1 alpha in Hedgehog stimulierten Adipozyten. Passend dazu war die zugehörige PDH Kinase überexpremiert in der cytoplasmatischen Fraktion reifer Fettzellen anzutreffen. Die glykolytischen Schlüsselenzyme Phosphofructokinase und Pyruvat Kinase M2 waren ebenfalls signifikant verändert in Hedgehog behandelten Proben. Diese Änderungen zeigten einen Glukosefluss in Richtung erhöhter Laktatproduktion. Detailuntersuchungen bestätigten Änderungen der Glykolyse und der mitochondrialen Funktion und zeigten auch erhöhte NAD+/NADH und NADP+/NADPH Ratios.Schlussendlich ermittelten diese (phospho)-proteomischen Daten Hh als einen unbekannten Regulator des Energiestoffwechsels, der zu einem erhöhten Glucosedurchsatz und Laktatproduktion führt.Hedgehog (Hh) signaling plays a fundamental role in developmental processes conserved from flies to humans, whereas aberrant Hh reactivation has been linked to cancer. Data from our laboratory identified (i) a key role for Hh in inhibiting white but not brown adipocyte differentiation in vitro and in vivo, and (ii) that this inhibitory Hh effect on white fat cell differentiation can be blocked by the activated T-cell cytokine interferon-gamma. However, no studies have so far addressed the impact of Hh on mature adipocytes and no data at all are available on Hh induced (phospho)-proteomic changes. Therefore, we set out to address both aspects and made use of quantitative proteomic approaches to identify new Hh targets and mechanisms in mature adipocytes.We performed 2D-PAGE and 1D-GeLC/MSMS with (i) phospho-protein enriched samples and (ii) cytoplasmic fractions to gain unbiased insight into rapid phosphorylation and lasting expression changes, respectively.Western blots and ELISA were performed to validate results. Phospho-protein-specific 1D- and 2D-gels revealed a highly reproducible phosphorylation of pyruvate dehydrogenase (PDH) E1 alpha in Hh treated samples. In line with this data we observed a significant up-regulation of PDH-kinase in the cytoplasmic fraction. Furthermore, the glycolytic key enzymes phosphofructokinase and pyruvate kinase M2 were significantly altered in Hh treated samples, leading to a shift towards lactate production. Pathway analyses identified major changes in glycolysis and mitochondrial function. Of note, increased NAD+/NADH and NADP+/NADPH ratios further reflected Hh induced metabolic reprogramming. In summary, our unbiased (phospho)-proteomic screen identified Hh as a hitherto unknown regulator of cellular energy metabolism, leading to increased glucose turnover and lactate production.<br /