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Microviridins are unique protease inhibitors from bloom-forming cyanobacteria that have both ecological and pharmacological relevance. Their peptide backbones are produced ribosomally, and ATP grasp ligases introduce omega-ester and omega-amide bonds to yield rare cage-like structures. Bioinformatic analysis of the microviridin biosynthesis gene cluster suggests a novel type of processing machinery, which could rationalize the challenging in vivo/in vitro reconstitution of the pathway. In this work, we report the establishment of a minimal expression system for microviridins. Through bioinformatics and mutational analysis of the MdnA leader peptide we identified and characterized a strictly conserved binding motif that is specific for microviridin ligases. Furthermore, we showed that the ABC transporter MdnE is crucial for cyclization and processing of microviridins and demonstrated that MdnE is essential for stability of the microviridin biosynthesis complex.
It is currently controversially discussed if the same freshwater microorganisms occur worldwide wherever their required habitats are realized, i.e., without any adaptation to local conditions below the species level. We performed laboratory experiments with flagellates and ciliates from three acidic mining lakes (AML, pH similar to 2.7) to investigate if similar habitats may affect similar organisms differently. Such man-made lakes provide suitable ecosystem models to test for the significance of strong habitat selection. To this end, we analyzed the growth response of three protist taxa (three strains of the phytoflagellate Chlamydomonas acidophila, two isolates of the phytoflagellate Ochromonas and two species of the ciliate genus Oxytricha) by exposing them to lake water of their origin and from the two other AML in a cross-factorial design. Population growth rates were measured as a proxy for their fitness. Results revealed significant effects of strain, lake (= habitat), and strain X habitat interaction. In the environmentally most adverse AML, all three protist taxa were locally adapted. In conclusion, our study demonstrates that (1) the same habitat may affect strains of the same species differently and that (2) similar habitats may harbor ecophysiologically different strains or species. These results contradict the 'everything is everywhere' paradigm.
Temperature-induced self-assembly of triple-responsive triblock copolymers in aqueous solutions
(2011)
A series of triple-thermoresponsive triblock copolymers from poly(N-n-propylacrylamide) (PNPAM, A), poly(methoxydiethylene glycol acrylate) (PMDEGA, B), and poly(N-ethylacrylamide) (PNEAM, C) was synthesized by sequential reversible addition-fragmentation chain transfer polymerizations. Polymers of differing block sequences, ABC, BAC, and ACB, with increasing phase transition temperatures in the order A < B < C were prepared. Their aggregation behavior in dilute aqueous solution was investigated using dynamic light scattering, turbidimetry, and NMR spectroscopy. The self-organization of such polymers was found to dependent strongly on the block sequence. While polymers with a terminal low-LCST (lower critical solution temperature) block undergo aggregation above the first phase transition temperature at 20-25 degrees C, triblock copolymers with the low-LCST block in the middle show aggregation only above the second phase transition. The collapse of the middle block is not sufficient to induce aggregation but produces instead stable, unimolecular micelles with a collapsed middle block, as supported by NMR and fluorescence probe data. Continued heating of all copolymers led to two additional thermal transitions at 40-55 and 70-80 degrees C, which could be correlated to the phase transitions of the B and C blocks, respectively. All polymers show a high tendency for cluster formation, once aggregation is induced. The carrier abilities of the triple responsive triblock copolymers for hydrophobic agents were probed with the solvatochromic fluorescence dye Nile Red. With passing through the first thermal transition, the block copolymers are capable of solubilizing Nile Red. In the case of block copolymers with sequences ABC or ACB, which bear the low-LCST block at one terminus, notable amounts of dye are solubilized already at this stage. In contrast, the hydrophobic probe is much less efficiently incorporated by the BAC triblock copolymer, which forms unimolecular micelles. Only after the collapse of the B block, when reaching the second phase transition at about 45 degrees C, does aggregation occur and solubilization becomes efficient. In the case of ABC and ACB polymers, the hydrophobic probe seems to partition between the originally collapsed A chains and the additional hydrophobic chains formed after the collapse of the less hydrophobic B block.
A set of double thermoresponsive diblock copolymers poly(N-n-propylacrylamide)-block-poly(N-ethylacrylamide) (PNPAM-b-PNEAM) was synthesised by sequential reversible addition-fragmentation chain transfer (RAFT) polymerisations. Using a twofold trimethylsilyl (TMS)-labeled RAFT-agent, the relative size of the two blocks was varied. While soluble as unimers below 15 degrees C, all copolymers exhibited thermally induced two-step self-assembly in water, due to distinct lower critical solution temperature (LCST) phase transitions of PNPAM (around 20 degrees C) and PNEAM (around 70 degrees C). Their temperature-dependent self-organisation in dilute aqueous solution was studied by turbidimetry, dynamic light scattering, transmission electron microscopy, and (1)H NMR spectroscopy. The copolymers show distinct, two-step self-organisation behaviour with respect to transition temperatures, aggregate type and size, which can be correlated to the relative lengths of the low and high LCST blocks. For polymers having short blocks with low LCST, the first thermal transition induces the formation of individual micelles. Further heating above the second thermal transition results reversibly either in a shrink of the micelle size or in aggregation of the micelles, with hydrodynamic diameters below 250 nm. In contrast in the case of polymers having a long block with low LCST, the first thermal transition already leads to clusters of micelles, while the second thermal transition makes the clusters shrink. Noteworthy, the twofold TMS-labeled end groups report not only on the molar masses of the polymers, but can simultaneously serve as NMR-probes for the self-assembly process. The signal of the TMS-aryl end group displays a reversible temperature dependent, two-step splitting that is indicative of the self-organisation of the block copolymers.
We propose a novel strategy for global sensitivity analysis of ordinary differential equations. It is based on an error-controlled solution of the partial differential equation (PDE) that describes the evolution of the probability density function associated with the input uncertainty/variability. The density yields a more accurate estimate of the output uncertainty/variability, where not only some observables (such as mean and variance) but also structural properties (e.g., skewness, heavy tails, bi-modality) can be resolved up to a selected accuracy. For the adaptive solution of the PDE Cauchy problem we use the Rothe method with multiplicative error correction, which was originally developed for the solution of parabolic PDEs. We show that, unlike in parabolic problems, conservation properties necessitate a coupling of temporal and spatial accuracy to avoid accumulation of spatial approximation errors over time. We provide convergence conditions for the numerical scheme and suggest an implementation using approximate approximations for spatial discretization to efficiently resolve the coupling of temporal and spatial accuracy. The performance of the method is studied by means of low-dimensional case studies. The favorable properties of the spatial discretization technique suggest that this may be the starting point for an error-controlled sensitivity analysis in higher dimensions.
Die Konsequenzen des Grollens in der arabischen Welt seit Anfang 2011 werden sich möglicherweise als weniger hilfreich für die Lösung des Nahostkonflikts erweisen als erhofft. Zumindest auf kurze Sicht hat sich die Nachbarschaft Israels in einen noch unberechenbareren und instabileren Ort verwandelt. Israel würde die Entstehung guten und repräsentativen Regierens in der arabischen Welt willkommen heißen. Doch sind die weiteren Entwicklungen in der Region kaum abzuschätzen.
In blue-light photoreceptors using flavin (BLUF), the signaling state is formed already within several 100 ps after illumination, with only small changes of the absorption spectrum. The accompanying structural evolution can, in principle, be monitored by femtosecond stimulated Raman spectroscopy (FSRS). The method is used here to characterize the excited-state properties of riboflavin and flavin adenine dinucleotide in polar solvents. Raman modes are observed in the range 90-1800 cm(-1) for the electronic ground state S-0 and upon excitation to the S-1 state, and modes >1000 cm(-1) of both states are assigned with the help of quantum-chemical calculations. Line shapes are shown to depend sensitively on resonance conditions. They are affected by wavepacket motion in any of the participating electronic states, resulting in complex amplitude modulation of the stimulated Raman spectra. Wavepackets in S-1 can be marked, and thus isolated, by stimulated-emission pumping with the picosecond Raman pulses. Excited-state absorption spectra are obtained from a quantitative comparison of broadband transient fluorescence and absorption. In this way, the resonance conditions for FSRS are determined. Early differences of the emission spectrum depend on excess vibrational energy, and solvation is seen as dynamic Stokes shift of the emission band. The ne state is evidenced only through changes of emission oscillator strength during solvation. S-1 quenching by adenine is seen with all methods in terms of dynamics, not by spectral intermediates.
Analysis of behavioural consistency is an important aspect of software engineering. In process and service management, consistency verification of behavioural models has manifold applications. For instance, a business process model used as system specification and a corresponding workflow model used as implementation have to be consistent. Another example would be the analysis to what degree a process log of executed business operations is consistent with the corresponding normative process model. Typically, existing notions of behaviour equivalence, such as bisimulation and trace equivalence, are applied as consistency notions. Still, these notions are exponential in computation and yield a Boolean result. In many cases, however, a quantification of behavioural deviation is needed along with concepts to isolate the source of deviation.
In this article, we propose causal behavioural profiles as the basis for a consistency notion. These profiles capture essential behavioural information, such as order, exclusiveness, and causality between pairs of activities of a process model. Consistency based on these profiles is weaker than trace equivalence, but can be computed efficiently for a broad class of models. In this article, we introduce techniques for the computation of causal behavioural profiles using structural decomposition techniques for sound free-choice workflow systems if unstructured net fragments are acyclic or can be traced back to S-or T-nets. We also elaborate on the findings of applying our technique to three industry model collections.
Process compliance measurement is getting increasing attention in companies due to stricter legal requirements and market pressure for operational excellence. In order to judge on compliance of the business processing, the degree of behavioural deviation of a case, i.e., an observed execution sequence, is quantified with respect to a process model (referred to as fitness, or recall). Recently, different compliance measures have been proposed. Still, nearly all of them are grounded on state-based techniques and the trace equivalence criterion, in particular. As a consequence, these approaches have to deal with the state explosion problem. In this paper, we argue that a behavioural abstraction may be leveraged to measure the compliance of a process log - a collection of cases. To this end, we utilise causal behavioural profiles that capture the behavioural characteristics of process models and cases, and can be computed efficiently. We propose different compliance measures based on these profiles, discuss the impact of noise in process logs on our measures, and show how diagnostic information on non-compliance is derived. As a validation, we report on findings of applying our approach in a case study with an international service provider.