The Alkylquinolone Repertoire of Pseudomonas aeruginosa is Linked to Structural Flexibility of the FabH‐like 2‐Heptyl‐3‐hydroxy‐4(1H)‐quinolone (PQS) Biosynthesis Enzyme PqsBC

Pseudomonas aeruginosa is a bacterial pathogen that causes life‐threatening infections in immunocompromised patients. It produces a large armory of saturated and mono‐unsaturated 2‐alkyl‐4(1H)‐quinolones (AQs) and AQ N‐oxides (AQNOs) that serve as signaling molecules to control the production of virulence factors and that are involved in membrane vesicle formation and iron chelation; furthermore, they also have, for example, antibiotic properties. It has been shown that the β‐ketoacyl‐acyl‐carrier protein synthase III (FabH)‐like heterodimeric enzyme PqsBC catalyzes the last step in the biosynthesis of the most abundant AQ congener, 2‐heptyl‐4(1H)‐quinolone (HHQ), by condensing octanoyl‐coenzyme A (CoA) with 2‐aminobenzoylacetate (2‐ABA), but the basis for the large number of other AQs/AQNOs produced by P. aeruginosa is not known. Here, we demonstrate that PqsBC uses different medium‐chain acyl‐CoAs to produce various saturated AQs/AQNOs and that it also biosynthesizes mono‐unsaturated congeners. Further, we determined the structures of PqsBC in four different crystal forms at 1.5 to 2.7 Å resolution. Together with a previous report, the data reveal that PqsBC adopts open, intermediate, and closed conformations that alter the shape of the acyl‐binding cavity and explain the promiscuity of PqsBC. The different conformations also allow us to propose a model for structural transitions that accompany the catalytic cycle of PqsBC that might have broader implications for other FabH‐enzymes, for which such structural transitions have been postulated but have never been observed.     

All-solid-state tunable continuous-wave ultraviolet source with high spectral purity and frequency stability

We present a novel approach for the generation of higly frequency-stable, widely tunable, single-frequency cw UV light that is suitable for high-resolution spectroscopy. Sum-frequency generation (SFG) of two solid-state sources with a single cavity resonant for both fundamental waves is employed. Using a highly stable, narrow-linewidth frequency-doubled cw Nd:YAG laser as a master laser and slaving to it the SFG cavity and the other fundamental wave from a Ti:sapphire laser, we generate UV radiation of 33-mW output power around 313 nm. Alternatively, we use a diode laser instead of the Ti:sapphire laser and produce an output power of 2.1 mW at 313 nm. With both setups we obtain a continuous tunability of >15 GHz, short-term frequency fluctuations in the submegahertz range, a long-term frequency drift below 100 MHz/h, and stable operation for several hours. The theory of optimized doubly resonant SFG is also given.     

Vector Image Segmentation by Piecewise Continuous Approximation

In this article we present an approach to the segmentation problem by a piecewise approximation of the given image with continuous functions. Unlike the common approach of Mumford and Shah in our formulation of the problem the number of segments is a parameter, which can be estimated. The problem can be stated as: Compute the optimal segmentation with a fixed number of segments, then reduce the number of segments until the segmentation result fulfills a given suitability. This merging algorithm results in a multi-objective optimization, which is not only resolved by a linear combination of the contradicting error functions. To constrain the problem we use a finite dimensional vector space of functions in our approximation and we restrict the shape of the segments. Our approach results in a multi-objective optimization: On the one hand the number of segments is to be minimized, on the other hand the approximation error should also be kept minimal. The approach is sound theoretically and practically: We show that for L ²-images a Pareto-optimal solution exists and can be computed for the discretization of the image efficiently.     

Elimination of artifacts due to occlusion and discretization problems in image space blurring techniques

Traditional computer graphics methods render images that appear sharp at all depths. Adding blur can add realism to a scene, provide a sense of scale, and draw a viewer’s attention to a particular region of a scene. Our image-based blur algorithm needs to distinguish whether a portion of an image is either from a single object or is part of more than one object. This motivates two approaches to identify objects after an image has been rendered. We illustrate how these techniques can be used in conjunction with our image space method to add blur to a scene.     

Treatment of insomnia. Getting to the root of sleeping problems

Insomnia may be periodic and transient, as caused by situational stress, or persistent, as caused by a chronic sleep disorder. Physicians can gain much information concerning the type, probable cause, onset, and duration of insomnia through history taking. A sleep diary may reveal helpful information, and input from the patient's sleeping partner can also be valuable. Complicating disorders, such as heart failure, prostatism, or depression, should be sought and specific treatment prescribed. Chemical dependency, too, requires appropriate treatment. These measures, institution of good sleep-hygiene practices, and behavior modification may resolve sleeplessness. The primary indication for use of hypnotic agents is transient sleep disruption caused by acute stress. When an agent is chosen, onset of action, metabolism, and side effects should be considered, especially in elderly patients. Addictive agents should not be given to patients with substance abuse problems. If insomnia persists, evaluation at a sleep-disorder center is recommended to facilitate design of an appropriate therapeutic regimen.     

Quantifying unfrozen water in frozen soil by high-field 2H NMR

To understand wintertime controls of biogeochemical processes in high latitude soils it is essential to distinguish between direct temperature effects and the effects of changes in water availability mediated by freezing. Efforts to separate these controls are hampered by a lack of adequate methods to determine the proportion of unfrozen water. In this study we present a high-field 2H2O NMR method for quantifying unfrozen water content in frozen soil. The experimental material consisted of the humic layer of a boreal spruce forest soil mixed with varying proportions of quartz sand and humidified with deuterium-enriched water. The relative standard deviation of unfrozen water content (measured as NMR signal integral) was less than 2% for repeated measurements on a given sample and 3.5% among all samples, based on a total of 16 measurements. As compared to 1H NMR, this 2H NMR method was found to be superior for several reasons: it is less sensitive to field inhomogeneity and paramagnetic impurities, it gives a bigger line shape difference between the ice and liquid signal, it shows a sharper response to water fusion, and it excludes the possibility of hydrogen in the organic material interfering with the measurement.     

Morphological image sequence processing

We present a morphological multi-scale method for image sequence processing, which results in a truly coupled spatio-temporal anisotropic diffusion. The aim of the method is not to smooth the level-sets of single frames but to denoise the whole sequence while retaining geometric features such as spatial edges and highly accelerated motions. This is obtained by an anisotropic spatio-temporal level-set evolution, where the additional artificial time variable serves as the multi-scale parameter. The diffusion tensor of the evolution depends on the morphology of the sequence, given by spatial curvatures of the level-sets and the curvature of trajectories (=acceleration) in sequence-time. We discuss different regularization techniques and describe an operator splitting technique for solving the problem. Finally we compare the new method with existing multi-scale image sequence processing methodologies.     

Prediction of work piece geometry in electrochemical cavity sinking

A computer-implemented model for predicting ECM work piece geometry has been developed and experimentally verified with a commercial ECM machine for cavity sinking in copper and 302-stainless steel with 2N K NO₃ electrolyte. Constant tool piece feed rates of 7–10 × 10^?4 cm/s, and applied voltages of 11–25 V were used. The model predicts the dependence of work piece geometry on operating conditions and on the electrochemical and physical properties of the metal—electrolyte pair. Comparison of eight equilibrium and six unsteady state experimental cavity profiles in copper showed satisfactory agreement with predictions, as did five equilibrium profiles for cavity sinking in 302-stainless steel.