Reciprocal regulation of PKA and rac signaling

Activated G protein-coupled receptors (GPCRs) and receptor tyrosine kinases relay extracellular signals through spatial and temporal controlled kinase and GTPase entities. These enzymes are coordinated by multifunctional scaffolding proteins for precise intracellular signal processing. The cAMP-dependent protein kinase A (PKA) is the prime example for compartmentalized signal transmission downstream of distinct GPCRs. A-kinase anchoring proteins tether PKA to specific intracellular sites to ensure precision and directionality of PKA phosphorylation events. Here, we show that the Rho-GTPase Rac contains A-kinase anchoring protein properties and forms a dynamic cellular protein complex with PKA. The formation of this transient core complex depends on binary interactions with PKA subunits, cAMP levels and cellular GTP-loading accounting for bidirectional consequences on PKA and Rac downstream signaling. We show that GTP-Rac stabilizes the inactive PKA holoenzyme. However, β-adrenergic receptor-mediated activation of GTP-Rac–bound PKA routes signals to the Raf-Mek-Erk cascade, which is critically implicated in cell proliferation. We describe a further mechanism of how cAMP enhances nuclear Erk1/2 signaling: It emanates from transphosphorylation of p21-activated kinases in their evolutionary conserved kinase-activation loop through GTP-Rac compartmentalized PKA activities. Sole transphosphorylation of p21-activated kinases is not sufficient to activate Erk1/2. It requires complex formation of both kinases with GTP-Rac1 to unleash cAMP-PKA–boosted activation of Raf-Mek-Erk. Consequently GTP-Rac functions as a dual kinase-tuning scaffold that favors the PKA holoenzyme and contributes to potentiate Erk1/2 signaling. Our findings offer additional mechanistic insights how β-adrenergic receptor-controlled PKA activities enhance GTP-Rac–mediated activation of nuclear Erk1/2 signaling

LPMLE3 : a novel 1-D approach to study water flow in streambeds using heat as a tracer

We introduce LPMLE3, a new 1-D approach to quantify vertical water flow components at streambeds using temperature data collected in different depths. LPMLE3 solves the partial differential equation for coupled water flow and heat transport in the frequency domain. Unlike other 1-D approaches it does not assume a semi-infinite halfspace with the location of the lower boundary condition approaching infinity. Instead, it uses local upper and lower boundary conditions. As such, the streambed can be divided into finite subdomains bound at the top and bottom by a temperature-time series. Information from a third temperature sensor within each subdomain is then used for parameter estimation. LPMLE3 applies a low order local polynomial to separate periodic and transient parts (including the noise contributions) of a temperature-time series and calculates the frequency response of each subdomain to a known temperature input at the streambed top. A maximum-likelihood estimator is used to estimate the vertical component of water flow, thermal diffusivity, and their uncertainties for each streambed subdomain and provides information regarding model quality. We tested the method on synthetic temperature data generated with the numerical model STRIVE and demonstrate how the vertical flow component can be quantified for field data collected in a Belgian stream. We show that by using the results in additional analyses, nonvertical flow components could be identified and by making certain assumptions they could be quantified for each subdomain. LPMLE3 performed well on both simulated and field data and can be considered a valuable addition to the existing 1-D methods

Die Jesuitenreduktionen in Paraguay im Lichte der kolonialethischen Diskussion

Leitmotiv der Arbeit ist die Frage nach dem Nächsten, dem Fremden, dem Unbekannten und dessen Wahrnehmung, die gerade in der heutigen Zeit nichts an ihrer Aktualität eingebüßt hat. Der Rahmen ist ein historischer, nämlich die Zeit der „Entgrenzung“ des geographischen Horizonts des Mittelalters und seine insbesondere in Spanien sich unmittelbar stellende Frage nach der Rechtmäßigkeit der Konquista, die sich in der Frage nach dem Nächsten und seinem Menschsein verdichtet. Die Reaktion darauf und die innovative Denkleistung von Vitoria bilden den Ausgangspunkt. Ferner wird auf den Zeitzeugen Las Casas und seinen Kampf für die Indios eingegangen. Dieser gipfelt in dem Disput von Valladolid mit seinem Hauptkontrahenten Sepúlveda. Breiten Raum nimmt das jesuitische Experiment in Paraguay, im speziellen die Guaraníreduktionen ein. Diese bilden die reale historische Grundlage für zahlreiche utopische europäische Projektionen. Es wird versucht, nach einer perspektivenreichen Durchleuchtung des Experiments zu einer abschließenden ethischen Würdigung zu kommen. In den Schlussgedanken wird der Kreis geschlossen und es finden aktuelle gegenwartsbezogene Bezüge statt.The study´s dominant leitmotif concerns the unknown neighbour, i.e. the stranger and how we perceive him, which is an issue much debated these days. The setting is historical, namely the period of a radical dissolution of boundaries and of expansion of Medieval geographical horizons. Closely connected to these phenomena of transition is the question of the legitimacy of the Spanish Conquista, an ethical dilemma which is of great urgency when considering the unknown neighbour as a fellow human being. Immediate responses to this challenging question and Vitoria´s innovative reflections provide the first impetus to the analysis. A further focus is on the contemporary eyewitness Las Casas and his struggles for the cause of the Indios, a conflict which culminates in the dispute of Valladolid with his main opponent, i.e. Sepúlveda. The Jesuit experiment in Paraguay, in particular the so-called Guaraní-reductions, is given expansive attention. These experiments are the historical and really existing origins of numerous European utopian fantasies and projections. It is the aim of the study to investigate the experiment from a plurality of perspectives and thus to reach conclusions which do justice to the ethical issues involved. The final section will close the argumentative circle and will attempt to refer to related current concerns

On the mechanism of RNA phosphodiester backbone cleavage in the absence of solvent

Ribonucleic acid (RNA) modifications play an important role in the regulation of gene expression and the development of RNA-based therapeutics, but their identification, localization and relative quantitation by conventional biochemical methods can be quite challenging. As a promising alternative, mass spectrometry (MS) based approaches that involve RNA dissociation in ‘top-down’ strategies are currently being developed. For this purpose, it is essential to understand the dissociation mechanisms of unmodified and posttranscriptionally or synthetically modified RNA. Here, we have studied the effect of select nucleobase, ribose and backbone modifications on phosphodiester bond cleavage in collisionally activated dissociation (CAD) of positively and negatively charged RNA. We found that CAD of RNA is a stepwise reaction that is facilitated by, but does not require, the presence of positive charge. Preferred backbone cleavage next to adenosine and guanosine in CAD of (M+nH)n+ and (M−nH)n− ions, respectively, is based on hydrogen bonding between nucleobase and phosphodiester moieties. Moreover, CAD of RNA involves an intermediate that is sufficiently stable to survive extension of the RNA structure and intramolecular proton redistribution according to simple Coulombic repulsion prior to backbone cleavage into c and y ions from phosphodiester bond cleavage

A power market-based operation support model for sub-daily hydropower regulation practices

AbstractWith increasing power production from renewable energy sources, sub-daily variations in energy demand need to be balanced. Today, hydropower is commonly used as balancing power. In this study, we quantified the impact of capacity constraints, in terms of reservoir volume and hydropower capacity, on the potential to comply with instant energy demand. To evaluate the impact, we developed two new metrics, power market impact and system efficiency ratio, which are based on two threshold flow regimes derived from natural flow as lower threshold release and regulated flow (based on hourly energy prices) as upper threshold release. The operation support model comprises 96 different regulation scenarios based on varying combinations of hydropower and reservoir capacities. For each scenario, an hourly water balance was simulated, to obtain the highest complying with upper threshold release based on actual energy demand. We tested the framework on the Kemijoki river with defined thresholds based on the natural flow regime (tributary river Ounasjoki) and the hourly energy price in Finland in 2017, and estimated the impact of regulation on hourly flow regime at the Taivalkoski hydropower station. The annual flow regime impact in 2013, 2014 and 2015 was estimated to be 74%, 84% and 61%, respectively, while the monthly impact varied from 27% to 100%. Our framework for evaluating interactions between the power market and sub-daily regulation practices is a useful novel tool for sustainable river management and can be easily applied to different rivers and regions and evaluated for different timescales.Abstract With increasing power production from renewable energy sources, sub-daily variations in energy demand need to be balanced. Today, hydropower is commonly used as balancing power. In this study, we quantified the impact of capacity constraints, in terms of reservoir volume and hydropower capacity, on the potential to comply with instant energy demand. To evaluate the impact, we developed two new metrics, power market impact and system efficiency ratio, which are based on two threshold flow regimes derived from natural flow as lower threshold release and regulated flow (based on hourly energy prices) as upper threshold release. The operation support model comprises 96 different regulation scenarios based on varying combinations of hydropower and reservoir capacities. For each scenario, an hourly water balance was simulated, to obtain the highest complying with upper threshold release based on actual energy demand. We tested the framework on the Kemijoki river with defined thresholds based on the natural flow regime (tributary river Ounasjoki) and the hourly energy price in Finland in 2017, and estimated the impact of regulation on hourly flow regime at the Taivalkoski hydropower station. The annual flow regime impact in 2013, 2014 and 2015 was estimated to be 74%, 84% and 61%, respectively, while the monthly impact varied from 27% to 100%. Our framework for evaluating interactions between the power market and sub-daily regulation practices is a useful novel tool for sustainable river management and can be easily applied to different rivers and regions and evaluated for different timescales

NALP3 inflammasome upregulation and CASP1 cleavage of the glucocorticoid receptor cause glucocorticoid resistance in leukemia cells

Glucocorticoids are universally used in the treatment of acute lymphoblastic leukemia (ALL), and resistance to glucocorticoids in leukemia cells confers poor prognosis. To elucidate mechanisms of glucocorticoid resistance, we determined the prednisolone sensitivity of primary leukemia cells from 444 patients newly diagnosed with ALL and found significantly higher expression of CASP1 (encoding caspase 1) and its activator NLRP3 in glucocorticoid-resistant leukemia cells, resulting from significantly lower somatic methylation of the CASP1 and NLRP3 promoters. Overexpression of CASP1 resulted in cleavage of the glucocorticoid receptor, diminished the glucocorticoid-induced transcriptional response and increased glucocorticoid resistance. Knockdown or inhibition of CASP1 significantly increased glucocorticoid receptor levels and mitigated glucocorticoid resistance in CASP1-overexpressing ALL. Our findings establish a new mechanism by which the NLRP3-CASP1 inflammasome modulates cellular levels of the glucocorticoid receptor and diminishes cell sensitivity to glucocorticoids. The broad impact on the glucocorticoid transcriptional response suggests that this mechanism could also modify glucocorticoid effects in other diseases

Spectral Decomposition of Regulatory Thresholds for Climate-Driven Fluctuations in Hydro- and Wind Power Availability

Abstract Climate-driven fluctuations in the runoff and potential energy of surface water are generally large in comparison to the capacity of hydropower regulation, particularly when hydropower is used to balance the electricity production from covarying renewable energy sources such as wind power. To define the bounds of reservoir storage capacity, we introduce a dedicated reservoir volume that aggregates the storage capacity of several reservoirs to handle runoff from specific watersheds. We show how the storage bounds can be related to a spectrum of the climate-driven modes of variability in water availability and to the covariation between water and wind availability. A regional case study of the entire hydropower system in Sweden indicates that the longest regulation period possible to consider spans from a few days of individual subwatersheds up to several years, with an average limit of a couple of months. Watershed damping of the runoff substantially increases the longest considered regulation period and capacity. The high covariance found between the potential energy of the surface water and wind energy significantly reduces the longest considered regulation period when hydropower is used to balance the fluctuating wind power

A power market-based operation support model for sub-daily hydropower regulation practices

Highlights • Investigate the impact of instant energy demand on sub-daily river regime. • Introducing power market impact index. • Introducing system efficiency ratio index. • Provides an efficient tool for sustainable river management. • Assess the interaction of power market and regulation practices.With increasing power production from renewable energy sources, sub-daily variations in energy demand need to be balanced. Today, hydropower is commonly used as balancing power. In this study, we quantified the impact of capacity constraints, in terms of reservoir volume and hydropower capacity, on the potential to comply with instant energy demand. To evaluate the impact, we developed two new metrics, power market impact and system efficiency ratio, which are based on two threshold flow regimes derived from natural flow as lower threshold release and regulated flow (based on hourly energy prices) as upper threshold release. The operation support model comprises 96 different regulation scenarios based on varying combinations of hydropower and reservoir capacities. For each scenario, an hourly water balance was simulated, to obtain the highest complying with upper threshold release based on actual energy demand. We tested the framework on the Kemijoki river with defined thresholds based on the natural flow regime (tributary river Ounasjoki) and the hourly energy price in Finland in 2017, and estimated the impact of regulation on hourly flow regime at the Taivalkoski hydropower station. The annual flow regime impact in 2013, 2014 and 2015 was estimated to be 74%, 84% and 61%, respectively, while the monthly impact varied from 27% to 100%. Our framework for evaluating interactions between the power market and sub-daily regulation practices is a useful novel tool for sustainable river management and can be easily applied to different rivers and regions and evaluated for different timescales

Distinct 5-methylcytosine profiles in poly(A) RNA from mouse embryonic stem cells and brain

Abstract Background Recent work has identified and mapped a range of posttranscriptional modifications in mRNA, including methylation of the N6 and N1 positions in adenine, pseudouridylation, and methylation of carbon 5 in cytosine (m5C). However, knowledge about the prevalence and transcriptome-wide distribution of m5C is still extremely limited; thus, studies in different cell types, tissues, and organisms are needed to gain insight into possible functions of this modification and implications for other regulatory processes. Results We have carried out an unbiased global analysis of m5C in total and nuclear poly(A) RNA of mouse embryonic stem cells and murine brain. We show that there are intriguing differences in these samples and cell compartments with respect to the degree of methylation, functional classification of methylated transcripts, and position bias within the transcript. Specifically, we observe a pronounced accumulation of m5C sites in the vicinity of the translational start codon, depletion in coding sequences, and mixed patterns of enrichment in the 3\u2032 UTR. Degree and pattern of methylation distinguish transcripts modified in both embryonic stem cells and brain from those methylated in either one of the samples. We also analyze potential correlations between m5C and micro RNA target sites, binding sites of RNA binding proteins, and N 6-methyladenosine. Conclusion Our study presents the first comprehensive picture of cytosine methylation in the epitranscriptome of pluripotent and differentiated stages in the mouse. These data provide an invaluable resource for future studies of function and biological significance of m5C in mRNA in mammals

Leitplanken gegen Studiengebühren und Bremer Finanzausgleichs-Tricksereien

Methylated cytosines in total poly(A) ESC RNA. (XLSX 761 kb