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To understand bacterial reactions to environmental stress or infection-related processes, it is necessary to identify the involved proteins. In mass spectrometry-based proteomics, the method of choice for spectra-to-peptide-match is database search, but in recent times, spectral libraries have come into focus. Here, we built a mass spectral library from Streptococcus pneumoniae D39, reflecting 76% of the theoretical proteome of the organism. Besides the proteins themselves, posttranslational protein modifications especially reveal central hubs of regulation in bacterial pathogenesis. Here, for example, phosphorylation events are involved in the signal transduction and regulation of virulence. Although there have been major advances in phosphoproteomics, identification of this modification is still challenging. To enhance the number of phosphorylated peptides, which can be reproducibly detected, a comprehensive mass spectral library of the S. pneumoniae D39 phosphoproteome has been compiled in addition to the comprehensive total proteome mass spectral library. The phosphopeptide library was manually validated, and the data quality was additionally proven by analyses of synthetic phosphorylated peptides. In total, 128 phosphorylated proteins were revealed, of which many are involved in glycolysis, purine metabolism, protein biosynthesis, and virulence. The publicly available, thoroughly validated spectral libraries are an excellent resource to improve and speed up future investigations on the proteome and phosphoproteome of pneumococci.

The increasing availability of sensor data at machine tools makes automatic chatter detection algorithms a trending topic in metal cutting. Two prominent and advanced methods for feature extraction via signal decomposition are Wavelet Packet Transform (WPT) and Ensemble Empirical Mode Decomposition (EEMD). We apply these two methods to time series acquired from an acceleration sensor at the tool holder of a lathe. Different turning experiments with varying dynamic behavior of the machine tool structure were performed. We compare the performance of these two methods with Support Vector Machine (SVM), Logistic Regression, Random Forest Classification and Gradient Boosting combined with Recursive Feature Elimination (RFE). We also show that the common WPT-based approach of choosing wavelet packets with the highest energy ratios as representative features for chatter does not always result in packets that enclose the chatter frequency, thus reducing the classification accuracy. Further, we test the transfer learning capability of each of these methods by training the classifier on one of the cutting configurations and then testing it on the other cases. It is found that when training and testing on data from the same cutting configuration both methods yield high accuracies reaching in one of the cases as high as 94% and 95%, respectively, for WPT and EEMD. However, our experimental results show that EEMD can outperform WPT in transfer learning applications with accuracy of up to 95%., Informative wavelet packet numbers were edited with respect to frequency ordering in section 3. Three more supervised learning algorithms were added to compare performance of both method (see section 5 and 6). For transfer learning, results of the application where classifiers are trained with two different stickout size data and tested on remaining cases were added into section 6.3

Additional file 1: Figure S1. PCA analysis. The RNA-sequencing results of the 3 biological replicates cultivated with glucose (G1-3) or ferulic acid (FA1-3) are depicted. Table S1. OD600 and HPLC values. Table S2. Sequencing statistics. Table S3. Gene expression values. Table S4. Regulated GO terms. Table S5. Regulated KEGG terms.

We show that the output of systems with time-varying delay can exhibit a new kind of chaotic behavior characterized by laminar phases, which are periodically interrupted by irregular bursts. Within each laminar phase the output intensity remains almost constant, but its level varies chaotically from phase to phase. In scalar systems the periodic dynamics of the lengths and the chaotic dynamics of the intensity levels can be understood and also tuned via two one-dimensional maps, which can be deduced from the nonlinearity of the delay equation and from the delay variation, respectively., Comment: 6 pages, 4 figures

Apart from some strongly fluorescent wood species, the general fluorescence of wood has long been ignored. Recent studies suggest that each species has a distinct fluorescence, originating from both basic components and characteristic extractives. However, wood colour and fluorescence rapidly change upon exposure to sunlight. In this study, 288 samples of Acer pseudoplatanus, Quercus robur, Picea abies and Juglans nigra were irradiated with different bands of ultraviolet (UV) and visible (VIS) light. Photosensitivity was examined in regards of colour, infrared absorbance (FTIR), and fluorescence imaging. UV light caused strong yellowing in all examined species, mostly correlating with lignin degradation, carbonyl formation and the appearance of a broad banded fluorescence emission. VIS light above 420 nm, however, caused different, partly contradicting effects in colour and fluorescence, and did not affect lignin. J. nigra proved to be most sensitive towards VIS-induced yellowing and bleaching. The main new finding of this study is that the native long wave fluorescence of wood was strongly decreased by VIS-irradiation above 510 nm wavelength in all samples. This effect was not species-specific, probably originating from a cross-species wood component. The results have potential impacts on non-destructive image-based evaluation methods and wood identification.

The cofactor F420 plays a central role as a hydride carrier in the primary and secondary metabolism of many bacterial and archaeal taxa. The cofactor is best known for its role in methanogenesis, where it facilitates thermodynamically difficult reactions. As the polyglutamate tail varies in length between different organisms, length profile analyses might be a powerful tool for distinguishing and characterizing different groups and pathways in various habitats. Here, the protocol describes the extraction and optimization of cofactor F420 detection by applying solid-phase extraction combined with high-performance liquid chromatography analysis independent of cultural or molecular biological approaches. The method was applied to gain additional information on the expression of cofactor F420 from microbial communities in soils, anaerobic sludge, and pure cultures and was evaluated by spiking experiments. Thereby, the study succeeded in generating different F420 tail-length profiles for hydrogenotrophic and acetoclastic methanogens in controlled methanogenic pure cultures as well as from environmental samples such as anaerobic digester sludge and soils.

Extraintestinal pathogenic Escherichia coli (ExPEC) strains constitute a serious and emerging clinical problem, as they cause a variety of infections and are usually highly antibiotic resistant. Many ExPEC strains are capable of evading the bactericidal effects of serum and causing sepsis. One critical factor for the development of septicemia is the increased serum survival (iss) gene, which is highly correlated with complement resistance and lethality. Although it is very important, the function of the iss gene has not been elucidated so far. We have been studying the serum survival of a septicemic strain of E. coli serotype O78, which has a group 4 capsule. Here, we show that the iss gene is required for the synthesis of capsules, which protect the bacteria from the bactericidal effect of complement. Moreover, we show that the deletion of the iss gene results in significantly increased binding of the complement proteins that constitute the membrane attack complex to the bacterial surface.

Objectives: To determine the effects of reorganizing a radiology institute from decentralized/modality-based to centralized/subspecialized radiology on radiologists, radiologic technicians, and referring physicians at a multi-center radiology network. Material and Methods: In 2017/2018 our multi-center radiology network was changed from decentralized/modality-based to centralized/subspecialized reporting. A survey was conducted among radiologists, technicians and two groups of referring physicians (main hospital and non-main hospitals). The following items were tested: Overall satisfaction, perceived quality of radiological reports, subjective productivity/efficiency, confidence of radiologists in their subspecialty, availability of radiologists and turnaround time. Two of five answering options on a 5-point Likert scale were considered to represent agreement. The Mann-Whitney-U-test served for statistical analyses in agreement before and after reorganization in each group. Results: For radiologists, a significant difference was observed in perceived quality of radiological reports 42/46 (91.3%) compared to 51/52 (98.1%; p = 0.013). For technicians, no significant differences were observed. In the group of main hospital referring physicians, significant differences were observed in overall satisfaction 129/152 (84.9%) compared to 164/174 (94.3%; p < 0.001) and in perceived quality of radiological reports 125/148 (72.8%) compared to 157/170 (92.4%; p = 0.001). In the group of non-main hospital referring physicians no significant differences were observed. Conclusion: The reorganization resulted in a significantly higher perceived quality of radiological reports for the groups of radiologists and main hospital referring physicians besides overall satisfaction for main hospital referring physicians. Specialized main hospital referring physicians value reports of specialized radiology, whereas less specialized, non-main hospital referring physicians did not experience any significant effect.

Extraintestinal pathogenic Escherichia coli (ExPEC) strains constitute a serious and emerging clinical problem, as they cause a variety of infections and are usually highly antibiotic resistant. Many ExPEC strains are capable of evading the bactericidal effects of serum and causing sepsis. One critical factor for the development of septicemia is the increased serum survival (iss) gene, which is highly correlated with complement resistance and lethality. Although it is very important, the function of the iss gene has not been elucidated so far. We have been studying the serum survival of a septicemic strain of E. coli serotype O78, which has a group 4 capsule. Here, we show that the iss gene is required for the synthesis of capsules, which protect the bacteria from the bactericidal effect of complement. Moreover, we show that the deletion of the iss gene results in significantly increased binding of the complement proteins that constitute the membrane attack complex to the bacterial surface.

In general we are interested in dynamical systems coupled to complex hysteresis. Therefore as a first step we investigated recently the dynamics of a periodically driven damped harmonic oscillator coupled to independent Ising spins in a random field. Although such a system does not produce hysteresis, we showed how to characterize the dynamics of such a piecewise-smooth system, especially in the case of a large number of spins [Zech, Otto, and Radons, Phys. Rev. E 101, 042217 (2020)2470-004510.1103/PhysRevE.101.042217]. In this paper we extend our model to spin dimers, thus pairwise interacting spins. We show in which cases two interacting spins can show elementary hysteresis, and we give a connection to the Preisach model, which allows us to consider an infinite number of spin pairs. This thermodynamic limit leads us to a dynamical system with an additional hysteretic force in the form of a generalized play operator. By using methods from general chaos theory, piecewise-smooth system theory, and statistics we investigate the chaotic behavior of the dynamical system for a few spins and also in the case of a larger number of spins by calculating bifurcation diagrams, Lyapunov exponents, fractal dimensions, and self-averaging properties. We find that the fractal dimensions and the magnetization are in general not self-averaging quantities. We show how the dynamical properties of the piecewise-smooth system for a large number of spins differs from the system in its thermodynamic limit.

[Image: see text] Staphylococcus aureus with spa-type t437 has been identified as a predominant community-associated methicillin-resistant S. aureus clone from Asia, which is also encountered in Europe. Molecular typing has previously shown that t437 isolates are highly similar regardless of geographical regions or host environments. The present study was aimed at assessing to what extent this high similarity is actually reflected in the production of secreted virulence factors. We therefore profiled the extracellular proteome, representing the main reservoir of virulence factors, of 20 representative clinical isolates by mass spectrometry. The results show that these isolates can be divided into three groups and nine subgroups based on exoproteome abundance signatures. This implies that S. aureus t437 isolates show substantial exoproteome heterogeneity. Nonetheless, 30 highly conserved extracellular proteins, of which about 50% have a predicted role in pathogenesis, were dominantly identified. To approximate the virulence of the 20 investigated isolates, we employed infection models based on Galleria mellonella and HeLa cells. The results show that the grouping of clinical isolates based on their exoproteome profile can be related to virulence. We consider this outcome important as our approach provides a tool to pinpoint differences in virulence among seemingly highly similar clinical isolates of S. aureus.

Phages are viruses that specifically infect and eventually kill their bacterial hosts. Bacterial fermentation and biotechnology industries see them as enemies, however, they are also investigated as antibacterial agents for the treatment or prevention of bacterial infections in various sectors. They also play key ecological roles in all ecosystems. Despite decades of research some aspects of phage biology are still poorly understood. In this study, we used label-free quantitative proteomics to reveal the proteotypes of Lactococcus lactis MG1363 during infection by the virulent phage p2, a model for studying the biology of phages infecting Gram-positive bacteria. Our approach resulted in the high-confidence detection and quantification of 59% of the theoretical bacterial proteome, including 226 bacterial proteins detected only during phage infection and 6 proteins unique to uninfected bacteria. We also identified many bacterial proteins of differing abundance during the infection. Using this high-throughput proteomic datasets, we selected specific bacterial genes for inactivation using CRISPR-Cas9 to investigate their involvement in phage replication. One knockout mutant lacking gene llmg_0219 showed resistance to phage p2 because of a deficiency in phage adsorption. Furthermore, we detected and quantified 78% of the theoretical phage proteome and identified many proteins of phage p2 that had not been previously detected. Among others, we uncovered a conserved small phage protein (pORFN1) coded by an unannotated gene. We also applied a targeted approach to achieve greater sensitivity and identify undetected phage proteins that were expected to be present. This allowed us to follow the fate of pORF46, a small phage protein of low abundance. In summary, this work offers a unique view of the virulent phages' takeover of bacterial cells and provides novel information on phage-host interactions.

We present a numerical approach to efficiently calculate spin-wave dispersions and spatial mode profiles in magnetic waveguides of arbitrarily shaped cross section with any non-collinear equilibrium magnetization which is translationally invariant along the waveguide. Our method is based on the propagating-wave dynamic-matrix approach by Henry et al. and extends it to arbitrary cross sections using a finite-element method. We solve the linearized equation of motion of the magnetization only in a single waveguide cross section which drastically reduces computational effort compared to common three-dimensional micromagnetic simulations. In order to numerically obtain the dipolar potential of individual spin-wave modes, we present a plane-wave version of the hybrid finite-element/boundary-element method by Frekdin and Koehler which, for the first time, we extend to a modified version of the Poisson equation. Our method is applied to several important examples of magnonic waveguides including systems with surface curvature, such as magnetic nanotubes, where the curvature leads to an asymmetric spin-wave dispersion. In all cases, the validity of our approach is confirmed by other methods. Our method is of particular interest for the study of curvature-induced or magnetochiral effects on spin-wave transport but also serves as an efficient tool to investigate standard magnonic problems.

We present a general approach for realizing safe collaborative behavior and high-accuracy operations in industrial robots. For example, collaborative behavior is required in human robot collaboration, whereas high-accuracy is needed in robotic machining. The operation sequence of a robot with corresponding often-contradictory demands on safety functions, control mode and accuracy is specified in a task-oriented path planner. The task planner generates optimized reference trajectories based on an environmental model, kinematic and task-specific constraints. The resulting reference path is fed into an online operation management, which monitors the workspace, ensures safety, and realizes different online control strategies especially for the dynamic tasks such as human robot collaboration or high-precision operations. For the high-accuracy tasks, we implemented a model predictive controller using a mechatronic model for predicting future configurations of the robot based on the measured present configuration. The model-predictive control is demonstrated for the compensation of deviations due to external forces that acts on the end effector of the robot.

Objectives The objective of this study was to compare the radiology report turnaround time (RTAT) between decentralized/modality-based and centralized/subspecialized radiological reporting at a multi-center radiology enterprise. Methods RTAT values for MRI, CT, and conventional radiography were compared between decentralized/modality-based (04 September 2017–22 December 2017) and centralized/subspecialized radiology (03 September 2018–21 December 2018) reporting grouped into three subspecializations (body radiology, musculoskeletal radiology, and neuroradiology) at eleven sites of a multi-center radiology enterprise. For the objective of this investigation, hospitals were defined as major and minor hospitals. The Mann-Whitney U test served for statistical analyses. Results Change of reporting system from decentralized/modality-based to centralized/subspecialized radiology resulted overall in a significant decrease of the RTAT: from 82 to 77 min for the first signature (p < 0.001), and 119 to 107 min and 295 to 238 min for the second signature (p < 0.001). Subgroup analyses demonstrate a significant decrease of the RTAT for MRI reports (e.g., second signature RTAT, 1051 to 401 min; p < 0.001) and conventional radiographs (e. g., second signature RTAT, 278 to 171 min; p < 0.001). The RTAT at major hospitals decreased from 288 to 245 min (second signature; p < 0.001) while the corresponding RTAT of minor hospitals decreased more remarkably, from 300 to 198 min (p < 0.001). However, the results were heterogenous; in some analyses, the RTAT even increased. The effect size analyses represent small effects. Conclusions Change of reporting system from decentralized/modality-based to centralized/subspecialized radiology was associated with a significant decreased RTAT. Specifically, the RTAT for MRI reports and conventional radiographs was significantly reduced. A pronounced RTAT decrease was observed at minor hospitals.

Our study presents new insights into the molecular mechanism that regulates the activity of SppA, a widely conserved bacterial membrane protease. We show that the membrane proteins SppA and SppI form a complex in the Gram-positive model bacterium B. subtilis and that SppI inhibits SppA protease activity in vitro and in vivo. Furthermore, we demonstrate that the C-terminal domain of SppI is involved in SppA inhibition. Since SppA, through its protease activity, contributes directly to resistance to lantibiotic peptides and cationic antibacterial peptides, we propose that the conserved SppA-SppI complex could play a major role in the evasion of bactericidal peptides, including those produced as part of human innate immune defenses., The membrane protease SppA of Bacillus subtilis was first described as a signal peptide peptidase and later shown to confer resistance to lantibiotics. Here, we report that SppA forms octameric complexes with YteJ, a membrane protein of thus-far-unknown function. Interestingly, sppA and yteJ deletion mutants exhibited no protein secretion defects. However, these mutant strains differed significantly in their resistance to antimicrobial peptides. In particular, sppA mutant cells displayed increased sensitivity to the lantibiotics nisin and subtilin and the human lysozyme-derived cationic antimicrobial peptide LP9. Importantly, YteJ was shown to antagonize SppA activity both in vivo and in vitro, and this SppA-inhibitory activity involved the C-terminal domain of YteJ, which was therefore renamed SppI. Most likely, SppI-mediated control is needed to protect B. subtilis against the potentially detrimental protease activity of SppA since a mutant overexpressing sppA by itself displayed defects in cell division. Altogether, we conclude that the SppA-SppI complex of B. subtilis has a major role in protection against antimicrobial peptides. IMPORTANCE Our study presents new insights into the molecular mechanism that regulates the activity of SppA, a widely conserved bacterial membrane protease. We show that the membrane proteins SppA and SppI form a complex in the Gram-positive model bacterium B. subtilis and that SppI inhibits SppA protease activity in vitro and in vivo. Furthermore, we demonstrate that the C-terminal domain of SppI is involved in SppA inhibition. Since SppA, through its protease activity, contributes directly to resistance to lantibiotic peptides and cationic antibacterial peptides, we propose that the conserved SppA-SppI complex could play a major role in the evasion of bactericidal peptides, including those produced as part of human innate immune defenses.

Presented at the 11th CIRP Conference on Photonic Technologies [LANE 2020] on September 7-10, 2020. © 2020 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/) Peer-review under responsibility of the Bayerisches Laserzentrum GmbH Abstract The presence of a gap (from tens to hundreds of microns) between two components may cause defects during laser-welding processes. Within this paper the process is investigated for two stainless steel plates in overlap configuration by means of multiphysyical CFD modeling. As a first step modeling results are compared with experimental data. Then the gap influence on the fluid dynamics and on the appearance of welding defects are analyzed. Based on the results adapted beam shapes are proposed that allow for an optimized energy distribution and thus an improved weld quality.

The lactococcal virulent phage p2 is a model for studying the Skunavirus genus, the most prevalent group of phages causing milk fermentation failures in cheese factories worldwide. This siphophage infects Lactococcus lactis MG1363, a model strain used to study Gram-positive lactic acid bacteria. The structural proteins of phage p2 have been thoroughly described, while most of its non-structural proteins remain uncharacterized. Here, we developed an integrative approach, making use of structural biology, genomics, physiology, and proteomics to provide insights into the function of ORF47, the most conserved non-structural protein of unknown function among the Skunavirus genus. This small phage protein, which is composed of three &alpha, helices, was found to have a major impact on the bacterial proteome during phage infection and to significantly reduce the emergence of bacteriophage-insensitive mutants.

The opportunistic pathogen

To understand bacterial reactions to environmental stress or infection-related processes, it is necessary to identify the involved proteins. In mass spectrometry-based proteomics, the method of choice for spectra-to-peptide-match is database search, but in recent times, spectral libraries have come into focus. Here, we built a mass spectral library from

We aim at an understanding of the dynamical properties of a periodically driven damped harmonic oscillator coupled to a Random Field Ising Model (RFIM) at zero temperature, which is capable to show complex hysteresis. The system is a combination of a continuous (harmonic oscillator) and a discrete (RFIM) subsystem, which classifies it as a hybrid system. In this paper we focus on the hybrid nature of the system and consider only independent spins in quenched random local fields, which can already lead to complex dynamics such as chaos and multistability. We study the dynamic behavior of this system by using the theory of piecewise-smooth dynamical systems and discontinuity mappings. Specifically, we present bifurcation diagrams, Lyapunov exponents as well as results for the shape and the dimensions of the attractors and the self-averaging behavior of the attractor dimensions and the magnetization. Furthermore we investigate the dynamical behavior of the system for an increasing number of spins and the transition to the thermodynamic limit, where the system behaves like a driven harmonic oscillator with an additional nonlinear smooth external force., Comment: https://journals.aps.org/pre/abstract/10.1103/PhysRevE.101.042217

Clostridioides difficile is the major pathogen causing diarrhea following antibiotic treatment. It is considered to be a strictly anaerobic bacterium, however, previous studies have shown a certain and strain-dependent oxygen tolerance. In this study, the model strain C. difficile 630Δerm was shifted to micro-aerobiosis and was found to stay growing to the same extent as anaerobically growing cells with only few changes in the metabolite pattern. However, an extensive change in gene expression was determined by RNA-Seq. The most striking adaptation strategies involve a change in the reductive fermentation pathways of the amino acids proline, glycine and leucine. But also a far-reaching restructuring in the carbohydrate metabolism was detected with changes in the phosphotransferase system (PTS) facilitated uptake of sugars and a repression of enzymes of glycolysis and butyrate fermentation. Furthermore, a temporary induction in the synthesis of cofactor riboflavin was detected possibly due to an increased demand for flavin mononucleotid (FMN) and flavin adenine dinucleotide (FAD) in redox reactions. However, biosynthesis of the cofactors thiamin pyrophosphate and cobalamin were repressed deducing oxidation-prone enzymes and intermediates in these pathways. Micro-aerobically shocked cells were characterized by an increased demand for cysteine and a thiol redox proteomics approach revealed a dramatic increase in the oxidative state of cysteine in more than 800 peptides after 15 min of micro-aerobic shock. This provides not only a catalogue of oxidation-prone cysteine residues in the C. difficile proteome but also puts the amino acid cysteine into a key position in the oxidative stress response. Our study suggests that tolerance of C. difficile towards O2 is based on a complex and far-reaching adjustment of global gene expression which leads to only a slight change in phenotype.

Recently, it was shown that certain systems with large time-varying delay exhibit different types of chaos, which are related to two types of time-varying delay: conservative and dissipative delays. The known high-dimensional Turbulent Chaos is characterized by strong fluctuations. In contrast, the recently discovered low-dimensional Laminar Chaos is characterized by nearly constant laminar phases with periodic durations and a chaotic variation of the intensity from phase to phase. In this paper we extend our results from our preceding publication [J. D. Hart, R. Roy, D. M\"uller-Bender, A. Otto, and G. Radons, PRL 123 154101 (2019)], where it is demonstrated that Laminar Chaos is a robust phenomenon, which can be observed in experimental systems. We provide a time-series analysis toolbox for the detection of robust features of Laminar Chaos. We benchmark our toolbox by experimental time series and time series of a model system which is described by a nonlinear Langevin equation with time-varying delay. The benchmark is done for different noise strengths for both the experimental system and the model system, where Laminar Chaos can be detected, even if it is hard to distinguish from Turbulent Chaos by a visual analysis of the trajectory., Comment: 16 pages, 14 figures; v2: minor corrections, typos; v3: minor corrections, typos, added journal reference and DOI

Aromatic compounds like phenyl acids derived from lignocellulose degradation have been suspected to negatively influence biogas production processes. However, results on this topic are still inconclusive. To study phenyl acid formation in batch reactors during the start-up phase of anaerobic degradation, different amounts of straw from grain were mixed with mesophilic and thermophilic sludge, respectively. Molecular biological parameters were assessed using next-generation sequencing and qPCR analyses. Metagenomic predictions were done via the program, piphillin. Methane production, concentrations of phenylacetate, phenylpropionate, phenylbutyrate, and volatile fatty acids were monitored chromatographically. Methanosarcina spp. was the dominant methanogen when high straw loads were effectively degraded, and thus confirmed its robustness towards overload conditions. Several microorganisms correlated negatively with phenyl acids, however, a negative effect, specifically on methanogens, could not be proven. A cascade-like increase/decrease from phenylacetate to phenylpropionate, and then to phenylbutyrate could be observed when methanogenesis was highly active. Due to these results, phenylacetate was shown to be an early sign for overload conditions, whereas an increase in phenylbutyrate possibly indicated a switch from degradation of easily available to more complex substrates. These dynamics during the start-up phase might be relevant for biogas plant operators using complex organic wastes for energy exploitation.

The strictly anaerobic bacterium C. difficile has become one of the most problematic hospital acquired pathogens and a major burden for health care systems. Although antibiotics work effectively in most C. difficile infections (CDIs), their detrimental effect on the intestinal microbiome paves the way for recurrent episodes of CDI. To develop alternative, non-antibiotics-based treatment strategies, deeper knowledge on the physiology of C. difficile, stress adaptation mechanisms and regulation of virulence factors is mandatory. The focus of this work was to tackle the thiol proteome of C. difficile and its stress-induced alterations, because recent research has reported that the amino acid cysteine plays a central role in the metabolism of this pathogen. We have developed a novel cysteine labeling approach to determine the redox state of protein thiols on a global scale. Applicability of this technique was demonstrated by inducing disulfide stress using the chemical diamide. The method can be transferred to any kind of redox challenge and was applied in this work to assess the effect of bile acids on the thiol proteome of C. difficile. We present redox-quantification for more than 1,500 thiol peptides and discuss the general difficulty of redox analyses of peptides possessing more than a single cysteine residue. The presented method will be especially useful not only when determining redox status, but also for providing information on protein quantity. Additionally, our comprehensive data set reveals protein cysteine sites particularly susceptible to oxidation and builds a groundwork for redox proteomics studies in C. difficile.

The process windows for welding Al and Cu are quite limited due to process instabilities like spiking or humping that lead to severe welding failures. Within this paper the fluiddynamical reasons for these instabilities will be analysed by means of multiphysical numerical simulations of the laser beam welding process. After a short introduction into the process model, the simulation results will be verified on basis of experimental results. Then we will focus on the analysis of the different process dynamics during welding for both Al and Cu. The reasons for the humping and spiking instabilities will be revealed. Finally an outlook on current developments, e.g. concerning the grain growth during solidification, will be given.

Numerous reviews have summarized the epidemiology, pathophysiology and the various therapeutic aspects of Coronavirus disease 2019 (COVID-19), but a practical guide on "how to treat whom with what and when" based on an understanding of the immunological background of the disease stages remains missing. This review attempts to combine the current knowledge about the immunopathology of COVID-19 with published evidence of available and emerging treatment options. We recognize that the information about COVID-19 and its treatment is rapidly changing, but hope that this guide offers those on the frontline of this pandemic an understanding of the host response in COVID-19 patients and supports their ongoing efforts to select the best treatments tailored to their patient's clinical status.

This work focuses on the wettability transition of nodular cast iron (NCI) modified using a femtosecond laser-produced micro/nano-structure. Laser parameters such as laser fluences (0.10 to 3.18 J/cm2), and scan speed (0.2 to 1.0 mm/s) were varied to realize hierarchical or dual-phase structures. Raman spectroscopy was performed on samples treated at 0.10 to 0.63 J/cm2 to investigate the mechanism involved in the disappearance of the graphite nodule with increasing laser fluence. Just-after the laser treatment, surface was superhydrophilic in nature and progressively became superhydrophobic after cleaning and subsequent application of vacuum pressure for 8 h. In the present investigation, a static contact angle (SCA) of 164 ± 4° was measured. The temperature-dependent durability of the superhydrophobic surface was analyzed by heating at a temperature range from 80 °C to 240 °C. The results showed a stable superhydrophobic characteristic up to 120 °C even after five cycles of repetition. Scanning electron microscope (SEM) analysis was conducted to capture the micrograph of structures. Moreover, the chemisorption mechanism during the vacuum process and over the period was analyzed using X-ray photoelectron spectroscopy (XPS) that indicated a higher concentration of non-polar functional groups on the surface could effectively increase the SCA.

Differential proteomics targeting the protein abundance is commonly used to follow changes in biological systems. Differences in localization and degree of post-translational modifications of proteins including phosphorylations are of tremendous interest due to the anticipated role in molecular regulatory processes. Because of their particular low abundance in prokaryotes, identification and quantification of protein phosphorylation is traditionally performed by either comparison of spot intensities on two-dimensional gels after differential phosphoprotein staining or gel-free by stable isotope labeling, sequential phosphopeptide enrichment and following LC-MS analysis. In the current work, we combined in a proof-of-principle experiment these techniques using 14 N/15 N metabolic labeling with succeeding protein separation on 2D gels. The visualization of phosphorylations on protein level by differential staining was followed by protein identification and determination of phosphorylation sites and quantification by LC-MS/MS. This approach should avoid disadvantages of traditional workflows, in particular the limited capability of peptide-based gel-free methods to quantify isoforms of proteins. Comparing control and stress conditions allowed for relative quantification in protein phosphorylation in Bacillus pumilus exposed to hydrogen peroxide. Altogether, we quantified with this method 19 putatively phosphorylated proteins.

Research organizations in the spatial sciences face a demanding and changing context, characterized by multiple tensions and significant uncertainty. Increasingly, researchers are also asked to provide policy advice through method-driven processes, rather than ad hoc. In this policy advice context, the paper focuses on a specific tension: strategic planning and developing policy-oriented projects. Until now, little is known about how non-university research organizations deal with this tension as regards providing policy advice in the spatial sciences. Therefore, the paper analyzes the strategy development of the Leibniz Institute of Ecological Urban and Regional Development (IOER) in Dresden. Both authors are affiliated to the IOER. Hence, the paper is written from an “insider perspective” based on scientific concepts such as strategic planning. The paper analyzes project examples that refer to climate change adaptation in cities and regions. It shows that challenges, aims and practices of policy...