Spermidine biases the resolution of Holliday junctions by phage λ integrase

Holliday junctions are a central intermediate in diverse pathways of DNA repair and recombination. The isomerization of a junction determines the directionality of the recombination event. Previous studies have shown that the identity of the central sequence of the junction may favor one of the two isomers, in turn controlling the direction of the pathway. Here we demonstrate that, in the absence of DNA sequence-mediated isomer preference, polycations are the major contributor to biasing strand cleavage during junction resolution. In the case of wild-type phage λ excision junctions, spermidine plays the dominant role in controlling the isomerization state of the junction and increases the rate of junction resolution. Spermidine also counteracts the sequence-imposed bias on resolution. The spermidine-induced bias is seen equally on supercoiled and linear excisive recombination junction intermediates, and thus is not just an artefact of in vitro recombination conditions. The contribution of spermidine requires the presence of accessory factors, and results in the repositioning of Int's core-binding domains on junctions, perhaps due to DNA-spermidine–protein interactions, or by influencing DNA conformation in the core region. Our results lead us to propose that spermidine together with accessory factors promotes the formation of the second junction isomer. We propose that this rearrangement triggers the activation of the second pair of Int active sites necessary to resolve Holliday junctions during phage λ Int-mediated recombination

Dunes in the world’s big rivers are characterized by low-angle lee-side slopes and a complex shape

Dunes are present in all the worlds’ big rivers and form critical agents of bedload transport, constitute appreciable sources of bed roughness and flow resistance, and generate stratification that is the most common depositional element of ancient alluvium. Yet our current models of dunes are conditioned by the geometry of bedforms observed in small rivers and laboratory experiments, and in which the downstream leeside angle is often assumed to be at the angle-of-repose. Here we show, using high-resolution bathymetry from a range of the worlds great rivers, that dunes are instead characterized predominantly by low-angle leeside slopes

Farewell, welfare state – hello, welfare regions? Chances and constraints of welfare management in the German federal system

The German welfare state is in crisis. Alarming long-term demographic trends, the still not fully digested consequences of German unification and the current economic downturn in much of the Eurozone have combined to create an urgent need for welfare reform. Yet the constitutional arrangements which govern the German political system, and well-entrenched political practice, mean that any such reform process is a daunting challenge. Thus, the welfare crisis is also a crisis of German-style co-operative federalism. Current empirical evidence makes for uncomfortable reading, and triggers debate on the nature of the German federation: have the two constitutional principles of federalism and establishing equal living conditions throughout the federation become mutually exclusive? However, as much of the welfare state is centred on the best utilisation of scarce financial resources, it is debatable to what extent alterations in the functional distribution of welfare responsibilities among the territorial levels of government can be regarded as a solution for the current problems. The article concludes that in the search for long-term sustainability of the welfare state the territorial dimension is likely to remain a secondary issue

Angiotensin II for the Treatment of Vasodilatory Shock

BACKGROUND Vasodilatory shock that does not respond to high-dose vasopressors is associated with high mortality. We investigated the effectiveness of angiotensin II for the treatment of patients with this condition. METHODS We randomly assigned patients with vasodilatory shock who were receiving more than 0.2 mu g of norepinephrine per kilogram of body weight per minute or the equivalent dose of another vasopressor to receive infusions of either angiotensin II or placebo. The primary end point was a response with respect to mean arterial pressure at hour 3 after the start of infusion, with response defined as an increase from baseline of at least 10 mm Hg or an increase to at least 75 mm Hg, without an increase in the dose of background vasopressors. RESULTS A total of 344 patients were assigned to one of the two regimens; 321 received a study intervention (163 received angiotensin II, and 158 received placebo) and were included in the analysis. The primary end point was reached by more patients in the angiotensin II group (114 of 163 patients, 69.9%) than in the placebo group (37 of 158 patients, 23.4%) (odds ratio, 7.95; 95% confidence interval [CI], 4.76 to 13.3; P<0.001). At 48 hours, the mean improvement in the cardiovascular Sequential Organ Failure Assessment (SOFA) score (scores range from 0 to 4, with higher scores indicating more severe dysfunction) was greater in the angiotensin II group than in the placebo group (-1.75 vs. -1.28, P = 0.01). Serious adverse events were reported in 60.7% of the patients in the angiotensin II group and in 67.1% in the placebo group. Death by day 28 occurred in 75 of 163 patients (46%) in the angiotensin II group and in 85 of 158 patients (54%) in the placebo group (hazard ratio, 0.78; 95% CI, 0.57 to 1.07; P = 0.12). CONCLUSIONS Angiotensin II effectively increased blood pressure in patients with vasodilatory shock that did not respond to high doses of conventional vasopressors. (Funded by La Jolla Pharmaceutical Company; ATHOS-3 ClinicalTrials.gov number, NCT02338843.)Peer reviewe

scRNA-Seq reveals distinct stem cell populations that drive hair cell regeneration after loss of Fgf and Notch signaling.

Loss of sensory hair cells leads to deafness and balance deficiencies. In contrast to mammalian hair cells, zebrafish ear and lateral line hair cells regenerate from poorly characterized support cells. Equally ill-defined is the gene regulatory network underlying the progression of support cells to differentiated hair cells. scRNA-Seq of lateral line organs uncovered five different support cell types, including quiescent and activated stem cells. Ordering of support cells along a developmental trajectory identified self-renewing cells and genes required for hair cell differentiation. scRNA-Seq analyses of fgf3 mutants, in which hair cell regeneration is increased, demonstrates that Fgf and Notch signaling inhibit proliferation of support cells in parallel by inhibiting Wnt signaling. Our scRNA-Seq analyses set the foundation for mechanistic studies of sensory organ regeneration and is crucial for identifying factors to trigger hair cell production in mammals. The data is searchable and publicly accessible via a web-based interface

Surviving Sepsis Campaign: International guidelines for management of severe sepsis and septic shock: 2008

SCOPUS: ar.jinfo:eu-repo/semantics/publishe

From Butler to Thornton: A Typology of Conflicting Readings of the Two Books of Scripture and Nature in the Church of England from the Eighteenth to the Twentieth Century

This dissertation studies the persistence within modern Anglicanism of (a) a classical "high view" of Scripture as the exemplar of creation, (b) the apologetic and phenomenological advantages of this view in the wake of Joseph Butler (1692-1752) due to (c) the kind of theodicy the Bible displays in contrast to more rationalistic proposals from deism to panentheism. (d) Finally, the view of theodicy and Scripture one takes will determine whether or not one reads the two books of Scripture and nature figurally. The "high view" was undercut by nineteenth-century sectarian polemics between Protestants and Tractarians. Yet a minority kept this tradition alive. Lionel Thornton (1884-1960) is important to this project because he escaped from Tractarian and Protestant dead ends. The dissertation lays out this broader Anglican story and then focuses on Thornton, for whose work I provide historical context and a detailed examination. I first analyze his early, philosophical-theological period where he defended an incarnational theodicy over against the panentheistic-monist alternative: the "soul-making" theodicy. Thornton's theodicy led him to take up a realist ("Platonist") metaphysic and phenomenology in order to resist the monist tendency to smooth over antinomies in Scripture and nature, especially the problem of evil. Next, I look at the theodical alternatives through the lens of Thornton and his mentor, John Neville Figgis (1866-1919). In contrast to the Modernist monism of Charles Raven and others, Figgis and Thornton resisted the temptation to offer an etiology of evil. They concluded that grace overcame evil by reordering the past. The last chapter, therefore, looks at Thornton’s view of the temporal and cosmic reach of Christ's reordering work of "recapitulation", and the hermeneutical consequences that follow: namely, that having reunited creation by rescuing it from the dispersive power of evil, every trivial detail of creation came to reflect Christ. I argue that this way of handling the Bible follows consistently from a biblical, non-monist, theodicy. Furthermore, I believe Thornton's project shared a family resemblance to Butler's, for, like the latter's, it indicated that the rejection of figural reading implied methodological atheism. This larger argument touches on contemporary hermeneutical debates within the Church.Doctor of Theology (ThD