MALDI analysis of Bacilli in spore mixtures by applying a quadrupole ion trap time-of-flight tandem mass spectrometer

A novel ion trap time-of-flight hybrid mass spectrometer (qIT-TOF MS) has been applied for peptide sequencing in proteolytic digests generated from spore mixtures of Bacilli. The method of on-probe solubilization and in situ proteolytic digestion of small, acid-soluble spore proteins has been recently developed in our laboratory, and microorganism identification in less than 20 min was accomplished. In this study, tryptic peptides were generated in situ from complex spore mixtures of B. subtilis 168, B. globigii, B. thuringiensis subs. Kurstaki, and B. cereus T, respectively. MALDI analysis of bacterial peptides generated was performed with an average mass resolving power of 6200 and a mass accuracy of up to 10 ppm using a trap-TOF tandem configuration. Precursor ions of interest were usually selected and stored in the quadrupole ion trap with their complete isotope distribution by choosing a window of +/- 2 Da. Sequence-specific information on isolated protonated peptides was gained via tandem MS experiments with an average mass resolving power of 4450 for product ion analysis, and protein and bacterial sources were identified by database searching.     

Characterization of Bacillus spore species and their mixtures using postsource decay with a curved-field reflectron

A strategy is proposed for the rapid identification of Bacillus spores, which relies on the selective release of a family of proteins, referred to as small, acid-soluble spore proteins (SASPs). In this work, SASPs were selectively solubilized from Bacillus spores on the MALDI sample plate by using 10% TFA. Proteolytic digests of SASPs generated in situ from spores of B. subtilis 168, B. globigii, B. thuringiensis subs. Kurstaki HD-1, B. cereus T, and the nonpathogenic strain B. anthracis Sterne were prepared in 5-25 min by using trypsin immobilized on Agarose beads and subsequently analyzed by MALDI-TOFMS using a curved-field reflectron. Protein identification was obtained by partial sequencing of distinctive tryptic peptides from Bacillus spores via post-source decay analysis combined with genome-based database searches by Mascot Sequence Query. Various unique SASPs were identified, allowing the characterization of Bacillus species by obtaining sequence-specific information on single peptides. The applicability of this approach for the rapid identification of Bacillus species was further established by analyzing spore mixtures.     

Microflow-based control of near-wall fluctuations for large viscous drag reduction

The stabilizing effect of microgroove surface morphology on viscous drag reduction was studied experimentally in the inlet region of a plane channel flow. The stabilization is thought to be due to the ability of a microgrooved surface pattern to suppress the velocity fluctuations in the spanwise direction on a restricted portion of the wetted surface, which prevents vorticity development at the wall and consequently across the entire flow field. This smart microflow control strategy, which works successfully only under very particular circumstances, was implemented in a microgroove-modified channel flow in which the front part has a microgrooved surface topology. The results of pressure drop measurements indicate that microgrooved surfaces can effectively stabilize laminar boundary layer development, leading to a significant reduction in the viscous drag. In the rear flat part of the microgroove-modified channel test section, a maximum drag reduction of DR @ 35%{\rm DR\simeq 35\%} was measured. This corresponds to an overall drag reduction of DR @ 16%{\rm DR\simeq 16\%} at a length Reynolds number of Re_x @ 10⁶.Re_x\simeq 10^6. The drag reduction effect persisted in a narrow range of flow velocities and for the reported experimental conditions corresponds to microgroove dimensions between 1.5 and 2.5 viscous length-scales.     

Elastic SI-Cache: consistent and scalable caching in multi-tier architectures

The new vision of cloud computing demands scalable, available and autonomic software platforms in order to deploy applications and services accessible anywhere and anytime. Multi-tier architectures are an important building block for many applications that are deployed in the cloud. This paper presents a novel caching and replication infrastructure that facilitates the scalable and elastic deployment of multi-tier architectures. Our Elastic SI-Cache is a novel multi-version cache that attains high performance and consistency in multi-tier systems. In contrast to most existing caches, Elastic SI-Cache provides snapshot isolation coherently across all tiers. Furthermore, Elastic SI-Cache supports scalable replication of the different tiers where replicas can be added or removed dynamically as needed, making the cache amenable for cloud computing environments. Elastic SI-Cache has been implemented and integrated into an open source JEE application server and its performance evaluated with the industrial benchmark SPECjAppServer.     

Discovery of interactive graphs for understanding and searching time-indexed corpora

Rich information spaces (like the Web or scientific publications) are full of “stories”: sets of statements that evolve over time, manifested as, for example, collections of news articles reporting events that relate to an evolving crime investigation, sets of news articles and blog posts accompanying the development of a political election campaign, or sequences of scientific papers on a topic. In this paper, we formulate the problem of discovering such stories as Evolutionary Theme Pattern Discovery, Summary and Exploration (ETP3). We propose a method and a visualisation tool for solving ETP3 by understanding, searching and interacting with such stories and their underlying documents. In contrast to existing approaches, our method concentrates on relational information and on local patterns rather than on the occurrence of individual concepts and global models. In addition, it relies on interactive graphs rather than natural language as the abstracted story representations. Furthermore, we present an evaluation framework. Two real-life case studies are used to illustrate and evaluate the method and tool.     

Detailed Structure–Function Correlations of Bacillus subtilis Acetolactate Synthase

Isobutanol is deemed to be a next‐generation biofuel and a renewable platform chemical. 1 Non‐natural biosynthetic pathways for isobutanol production have been implemented in cell‐based and in vitro systems with Bacillus subtilis acetolactate synthase (AlsS) as key biocatalyst. 2 – 6 AlsS catalyzes the condensation of two pyruvate molecules to acetolactate with thiamine diphosphate and Mg²⁺ as cofactors. AlsS also catalyzes the conversion of 2‐ketoisovalerate into isobutyraldehyde, the immediate precursor of isobutanol. Our phylogenetic analysis suggests that the ALS enzyme family forms a distinct subgroup of ThDP‐dependent enzymes. To unravel catalytically relevant structure‐function relationships, we solved the AlsS crystal structure at 2.3 Å in the presence of ThDP, Mg²⁺ and in a transition state with a 2‐lactyl moiety bound to ThDP. We supplemented our structural data by point mutations in the active site to identify catalytically important residues.     

Holzbrücken – Erkenntnisse zum Feuchteverhalten in den Tragquerschnitten aus Langzeitmessungen

Timber bridges – Long term monitoring results of moisture diffusion in the load carrying cross‐sections Timber road bridges have been built worldwide for centuries. The high performance of wood is approved through many constructions. However, there are still doubts about using wood by the planning engineer, which reduce the number of realized projects. The moisture induced stresses in cross‐sections according to the varied ambient climate are mainly investigated under laboratory conditions. The results show that the moisture content changes differently over the cross‐section and leads to moisture induced stresses. The discussion about methods for the assessment of the moisture induced stresses in timber bridge cross‐ections is continuing. Results observed in the long term monitoring of seven timber bridges provide first guidelines for practitioners. For all constructions, the moisture content was measured regarding the ambient micro climate. As result, the influences according to the ambient micro climate are not insignificant. Positive is that the directly measured moisture content is less compared to the equilibrium moisture content calculated from the data of a close by meteorological station. The analysis of the moisture gradient over the cross‐section allows the definition of an active or passive zone.     

Hydroxyl radical production via the photo-Fenton reaction in the presence of fulvic acid

The photo-Fenton reaction, the reaction of photoproduced Fe(II) with H2O2 to form the highly reactive hydroxyl radical (OH*), could be an important source of OH* in sunlit natural waters. To determine if the photo-Fenton reaction is significant in mildly acidic surface waters, we conducted experiments simulating conditions representative of natural freshwaters using solutions of standard fulvic acid and amorphous iron oxide at pH 6.0. A probe method measuring 14CO2 produced by the reaction of 14C-labeled formate with OH* was used to detect small OH* production rates without otherwise influencing the chemical reactions occurring in the experiments. Net H2O2 accumulation was simultaneously measured using an acridinium ester chemiluminescence method. Measured losses of H2O2 by reaction with Fe(II) in dark experiments produced approximately the expected quantities of OH*. The difference between H2O2 accumulation in the presence and absence of Fe(III) in fulvic acid solutions exposed to light was interpreted as the loss of H2O2 by reaction with photoproduced Fe(II), consistent with measured OH* production rates. The Fe ligand desferrioxamine mesylate eliminated both OH* production and H2O2 photoloss induced by Fe. Our results imply that when Fe is a major sink of H2O2, the photo-Fenton reaction is likely to be the most important source of OH*, leading to a significant sink of organic compounds in a wide variety of sunlit freshwaters.     

Indications for the applicability of element signature analysis for the determination of the geographic origin of dried beef and poultry meat

In order to determine the geographic origin of poultry and dried beef, concentrations of a total of 72 different elements (occasionally represented with several isotopes) were analyzed by inductively coupled plasma high resolution mass spectrometry (ICP-HRMS). Additionally, gross chemical composition (GCC) was analyzed. The 25 poultry breast filets samples originated from Switzerland, France, Germany, Hungary, Brazil, and Thailand, and the 23 dried beef samples, made from M. biceps femoris and M. semitendinosus, were produced in Switzerland, Austria, Australia, United States, and Canada out of raw meat originating either from these or from other countries. A total of 66 and 46 of the elements and isotopes followed were detected in beef and poultry, respectively. For statistical analyses, only the most abundant isotopes per element were used. For both poultry meat and dried beef, a differentiation of the origins was possible using those elements, which were significantly different across countries (As, Na, Rb, and Tl in poultry; B, Ca, Cd, Cu, Dy, Eu, Ga, Li, Ni, Pd, Rb, Sr, Te, Tl, Tm, V, Yb, and Zn in beef). No sufficient differentiation between origins was possible with GCC. Further studies have to confirm the suitability of this approach for meat authentication with more samples.     

APPLICATION OF CONCENTRATED SOLAR RADIATION TO HIGH TEMPERATURE DETOXIFICATION AND RECYCLING PROCESSES OF HAZARDOUS WASTES

In many cases, hazardous wastes are subject to thermal treatment at elevated temperatures. Some types of wastes do not have a sufficient calorific value to cover the heat demand of the high temperature process. For thermal treatment of e.g. filter residues, dusts, sulfuric acid, aluminium dross, foundry sand, or waste water, supplementary energy supply is needed. The specific energy demand ranges from 0.5 to 2.5 kWh/kg (2–10 MJ/kg). An important aim of process optimisation is the reduction of (fossil) energy consumption and exhaust gas flow. Concentrated solar energy promises advantages when applied to high energy consuming waste treatment processes with regard to substitute fossil or electric energy consumption, to reduce CO₂ emissions, and exhaust gas flow. In parallel to conceptional studies, a solar-heated rotary kiln mini-plant has been designed and constructed for tests in the DLR solar furnace. The tests will give indications of boundary conditions for solar thermal treatment or conversion of selected hazardous materials.