Osmotic stress-dependent serine phosphorylation of the histidine kinase homologue DokA

BACKGROUND: Two-component systems consisting of histidine kinases and their corresponding receivers are widespread in bacterial signal transduction. In the past few years, genes coding for homologues of two-component systems were also discovered in eukaryotic organisms. DokA, a homologue of bacterial histidine kinases, is an element of the osmoregulatory pathway in the amoeba Dictyostelium. The work described here addresses the question whether DokA is phosphorylated in vivo in response to osmotic stress. RESULTS: We have endogenously overexpressed individual domains of DokA to investigate post-translational modification of the protein in response to osmotic shock in vivo. Dictyostelium cells were labeled with [(32)P]-orthophosphate, exposed to osmotic stress and DokA fragments were subsequently isolated by immunoprecipitation. Thus, a stress-dependent phosphorylation could be demonstrated, with the site of phosphorylation being located in the kinase domain. We demonstrate biochemically that the phosphorylated amino acid is serine, and by mutational analysis that the phosphorylation reaction is not due to an autophosphorylation of DokA. Furthermore, mutation of the conserved histidine did not affect the osmostress-dependent phosphorylation reaction. CONCLUSIONS: A stimulus-dependent serine phosphorylation of a eukaryotic histidine kinase homologue was demonstrated for the first time in vivo. That implies that DokA, although showing typical structural features of a bacterial two-component system, might be part of a eukaryotic signal transduction pathway that involves serine/threonine kinases

Realizing Hopf Insulators in Dipolar Spin Systems

The Hopf insulator represents a topological state of matter that exists outside the conventional ten-fold way classification of topological insulators. Its topology is protected by a linking number invariant, which arises from the unique topology of knots in three dimensions. We predict that three-dimensional arrays of driven, dipolar-interacting spins are a natural platform to experimentally realize the Hopf insulator. In particular, we demonstrate that certain terms within the dipolar interaction elegantly generate the requisite non-trivial topology, and that Floquet engineering can be used to optimize dipolar Hopf insulators with large gaps. Moreover, we show that the Hopf insulator's unconventional topology gives rise to a rich spectrum of edge mode behaviors, which can be directly probed in experiments. Finally, we present a detailed blueprint for realizing the Hopf insulator in lattice-trapped ultracold dipolar molecules; focusing on the example of ${}^{40}$K$^{87}$Rb, we provide quantitative evidence for near-term experimental feasibility.Comment: 6 + 7 pages, 3 figure

Coping strategies in anxious surgical patients

Multiple hierarchical regression analysis. This supplement shows a table summarizing results of the multiple hierarchical regression analysis concerning the contribution of various patient characteristics to the prediction of surgical and anaesthesia anxiety. (DOCX 39 kb

Optimization approach for the combined planning and control of an agile assembly system for electric vehicles

For some years now, the automotive industry has been challenged by growing market dynamics, shorter product lifecycles and customers' increasing demands for individualization. In order to cope with this development, the automotive assembly needs to adapt quickly to changing demands with a low level of investment in the future. Under the current circumstances, the traditional line assembly for high volume production is reaching its limits in terms of adaptability and scalability. A promising solution to address the current challenges is the concept of the agile assembly. The concept of agile assembly breaks up the rigid linkage of assembly stations and, thus, enables full flexibility in the sequence of assembly operations only limited by the precedence graph. Therefore, the routing of electric vehicles in the agile assembly is based on the availability of resources such as assembly stations and automated guided vehicles that handle the material supply. Further, by transferring the transport function to the vehicle itself, investments for convey or systems are eliminated. This research work presents an optimization approach for the machine scheduling and transportation planning, which derives instructions for electric vehicles, assembly stations as well as automated guided vehicles. For each electric vehicle, an optimized route is calculated, taking into account product-specific precedence graphs and minimizing the overall makespan. In addition, the machine scheduling and transportation planning is integrated into a combined planning and control concept which covers the allocation of resources and the assignment of capabilities of the entire assembly system. The approach is implemented and applied to a practical case of a compact electric vehicle. Thus, the work contributes to the evaluation of agile assembly systems in automotive production

Identification and simulation of surface alpha events on passivated surfaces of germanium detectors and the influence of metalisation

Events from alpha interactions on the surfaces of germanium detectors are a major contribution to the background in germanium-based searches for neutrinoless double-beta decay. Surface events are subject to charge trapping, affecting their pulse shape and reconstructed energy. A study of alpha events on the passivated end-plate of a segmented true-coaxial n-type high-purity germanium detector is presented. Charge trapping is analysed in detail and an existing pulse-shape analysis technique to identify alpha events is verified with mirror pulses observed in the non-collecting channels of the segmented test detector. The observed radial dependence of charge trapping confirms previous results. A dependence of the probability of charge trapping on the crystal axes is observed for the first time. A first model to describe charge trapping effects within the framework of the simulation software SolidStateDetectors.jl is introduced. The influence of metalisation on events from low-energy gamma interactions close to the passivated surface is also presented

Simultaneous MMC readout using a tailored {\mu}MUX based readout system

Magnetic microcalorimeters (MMCs) are cryogenic, energy-dispersive single-particle detectors providing excellent energy resolution, intrinsically fast signal rise time, quantum efficiency close to 100\%, large dynamic range as well as almost ideal linear response. One of the remaining challenges to be overcome to ultimately allow for the utilization of large-scale MMC based detector arrays with thousands to millions of individual pixels is the realization of a SQUID based multiplexing technique particularly tailored for MMC readout. Within this context, we report on the first truly multiplexed readout of an MMC based detector array using a frequency-division multiplexing approach realized by a custom microwave SQUID multiplexer based readout system.Comment: Conference: ASC2022 (accepted for publication in IEEE Transactions on Applied Superconductivity

Temperature Dependence of the Electron-Drift Anisotropy and Implications for the Electron-Drift Model

The electron drift in germanium detectors is modeled making many assumptions. Confronted with data, these assumptions have to be revisited. The temperature dependence of the drift of electrons was studied in detail for an n-type segmented point-contact germanium detector. The detector was mounted in a temperature controlled, electrically cooled cryostat. Surface events were induced with collimated 81 keV photons from a $^{133}$Ba source. A detailed analysis of the rise time of pulses collected in surface scans, performed at different temperatures, is presented. The longitudinal anisotropy of the electron drift decreases with rising temperature. A new approach, making use of designated rise-time windows determined by simulations using SolidStateDetectors.jl, was used to isolate the longitudinal drift of electrons along different axes to quantify this observation. The measured temperature dependence of the longitudinal drift velocities combined with the standard electron drift model as widely used in relevant simulation packages results in unphysical predictions. A first suggestion to modify the electron-drift model is motivated and described. The results of a first implementation of the modified model in SolidStateDetectors.jl are shown. They describe the data reasonably well. A general review of the model and the standard input values for mobilities is suggested

Integrated Strategy for Sustainable Cattle Fever Tick Eradication in USA is Required to Mitigate the Impact of Global Change

The ticks Rhipicephalus (Boophilus) annulatus and R. (B.) microplus, commonly known as cattle and southern cattle tick, respectively, impede the development and sustainability of livestock industries throughout tropical and other world regions. They affect animal productivity and wellbeing directly through their obligate blood-feeding habit and indirectly by serving as vectors of the infectious agents causing bovine babesiosis and anaplasmosis. The monumental scientific discovery of certain arthropod species as vectors of infectious agents is associated with the history of research on bovine babesiosis and R. annulatus. Together, R. microplus and R. annulatus are referred to as cattle fever ticks (CFT). Bovine babesiosis became a regulated foreign animal disease in the United States of America (U.S.) through efforts of the Cattle Fever Tick Eradication Program (CFTEP) established in 1906. The U.S. was declared free of CFT in 1943, with the exception of a permanent quarantine zone in south Texas along the border with Mexico. This achievement contributed greatly to the development and productivity of animal agriculture in the U.S. The permanent quarantine zone buffers CFT incursions from Mexico where both ticks and babesiosis are endemic. Until recently, the elimination of CFT outbreaks relied solely on the use of coumaphos, an organophosphate acaricide, in dipping vats or as a spray to treat livestock, or the vacation of pastures. However, ecological, societal, and economical changes are shifting the paradigm of systematically treating livestock to eradicate CFT. Keeping the U.S. CFT-free is a critical animal health issue affecting the economic stability of livestock and wildlife enterprises. Here, we describe vulnerabilities associated with global change forces challenging the CFTEP. The concept of integrated CFT eradication is discussed in reference to global change

Hsa-miR-375 is a predictor of local control in early stage breast cancer

Background: A long-term analysis by the Early Breast Cancer Trialist Group (EBCTG) revealed a strong correlation between local control and cancer-specific mortality. MicroRNAs (miRs), short (20-25 nucleotides) non-coding RNAs, have been described as prognosticators and predictors for breast cancer in recent years. The aim of the current study was to identify miRs that can predict local control after breast conserving therapy (BCT) in early stage breast cancer. Results: Clinical data of 46 early stage breast cancer patients with local relapse after BCT were selected from the institutional database. These patients were matched to 101 control patients showing identical clinical features but without local relapse. The study was conducted in two steps. (1) In the pilot study, 32 patients (16 relapses versus 16 controls) were screened for the most de-regulated microRNAs (= candidate microRNAs) in a panel of 1250 miRs by microarray technology. Eight miRs were found to be significantly de-regulated. (2) In the validation study, the candidate microRNAs were analyzed in an independent cohort of 115 patients (30 relapses versus 85 controls) with reverse transcription quantitative polymerase chain reaction (RT-qPCR). From these eight candidates, hsa-miR-375 could be validated. Its median fold change was 2.28 (Mann-Whitney U test, corrected p value = 0.008). In the log-rank analysis, high expression levels of hsa-miR-375 correlated with a significantly higher risk of local relapse (p = 0.003). In a multivariate analysis (forward stepwise regression) including established predictors and prognosticators, hsa-miR-375 was the only variable that was able to distinguish the statistical significance between relapse and control groups (raw p value = 0.000195 HR = 0.76, 95 % CI 0.66-0.88;corrected p value = 0.005). Conclusions: Hsa-miR-375 predicts local control in patient with early stage breast cancer, especially in estrogen receptor alpha (ER-alpha)-positive patients. It can therefore serve as an additional molecular marker for treatment choice independently from known predictors and prognosticators. Validation in larger prospective studies is warranted