Adverse environmental conditions are detrimental to plant growth and development. Acclimation to abiotic stress conditions involves activation of signaling pathways which often results in changes in gene expression via networks of transcription factors (TFs). Mediator is a highly conserved co-regulator complex and an essential component of the transcriptional machinery in eukaryotes. Some Mediator subunits have been implicated in stress-responsive signaling pathways; however, much remains unknown regarding the role of plant Mediator in abiotic stress responses. Here, we use RNA-seq to analyze the transcriptional response of Arabidopsis thaliana to heat, cold and salt stress conditions. We identify a set of common abiotic stress regulons and describe the sequential and combinatorial nature of TFs involved in their transcriptional regulation. Furthermore, we identify stress-specific roles for the Mediator subunits MED9, MED16, MED18 and CDK8, and putative TFs connecting them to different stress signaling pathways. Our data also indicate different modes of action for subunits or modules of Mediator at the same gene loci, including a co-repressor function for MED16 prior to stress. These results illuminate a poorly understood but important player in the transcriptional response of plants to abiotic stress and identify target genes and mechanisms as a prelude to further biochemical characterization.

Signaling pathways in biological systems rely on specific interactions between multiple biomolecules. Fluorescence fluctuation spectroscopy provides a powerful toolbox to quantify such interactions directly in living cells. Cross-correlation analysis of spectrally separated fluctuations provides information about intermolecular interactions but is usually limited to two fluorophore species. Here, we present scanning fluorescence spectral correlation spectroscopy (SFSCS), a versatile approach that can be implemented on commercial confocal microscopes, allowing the investigation of interactions between multiple protein species at the plasma membrane. We demonstrate that SFSCS enables cross-talk-free cross-correlation, diffusion, and oligomerization analysis of up to four protein species labeled with strongly overlapping fluorophores. As an example, we investigate the interactions of influenza A virus (IAV) matrix protein 2 with two cellular host factors simultaneously. We furthermore apply raster spectral image correlation spectroscopy for the simultaneous analysis of up to four species and determine the stoichiometry of ternary IAV polymerase complexes in the cell nucleus.

The mechanical muscular oscillations are rarely the objective of investigations regarding the identification of a biomarker for Parkinson's disease (PD). Therefore, the aim of this study was to investigate whether or not this specific motor output differs between PD patients and controls. The novelty is that patients without tremor are investigated performing a unilateral isometric motor task. The force of armflexors and the forearm acceleration (ACC) were recorded as well as the mechanomyography of the biceps brachii (MMGbi), brachioradialis (MMGbra) and pectoralis major (MMGpect) muscles using a piezoelectric-sensor-based system during a unilateral motor task at 70% of the MVIC. The frequency, a power-frequency-ratio, the amplitude variation, the slope of amplitudes and their interlimb asymmetries were analysed. The results indicate that the oscillatory behavior of muscular output in PD without tremor deviates from controls in some parameters: Significant differences appeared for the power-frequency-ratio (p=0.001, r=0.43) and for the amplitude variation (p=0.003, r=0.34) of MMGpect. The interlimb asymmetries differed significantly concerning the power-frequency-ratio of MMGbi (p=0.013, r=0.42) and MMGbra (p=0.048, r=0.39) as well as regarding the mean frequency (p=0.004, r=0.48) and amplitude variation of MMGpect (p=0.033, r=0.37). The mean (M) and variation coefficient (CV) of slope of ACC differed significantly (M: p=0.022, r=0.33; CV: p=0.004, r=0.43). All other parameters showed no significant differences between PD and controls. It remains open, if this altered mechanical muscular output is reproducible and specific for PD.

The quality of the reference signal is essential for the adaptation process of an LMS or one of its derivatives. The reference signal affects the stability, the convergence rate and the maximum achievable attenuation. Since the error signal and the control signal are available as numerical values in the algorithm for the LMS, the reference signal can be calculated from both signals. The error signal is the interference between the control signal and the reference signal. This interference of the control signal and the reference signal can be noted mathematically as a simple addition. It is therefore possible to deduce the reference signal from a known error signal and control signal. This approach is the basis of the generated-x LMS (GxLMS) developed by us. It calculates the reference signal itself without having to rely on an externally supplied reference signal. The advantages of the GxLMS are primarily in fields where the reference signal is difficult or impossible to detect. For example, the detection of the reference signal can be problematic due to design reasons or measurement technology. For example, flow noise could have a negative effect on an acoustic detection of the reference signal. However, the calculation of the reference signal in the GxLMS represents a further feedback signal path, which affects the stability of the algorithm as a whole. Based on the theoretical principles mathematically sufficient convergence conditions can be formulated taking into account the delays existing in the signal paths. The experimental testing took place on an acoustic duct with monofrequency disturb signals. Since the use of an efficient design of experiments (DoE) could be excluded, the measurement was designed as parameter variation (one factor at time) and therefore very time-consuming. The theoretical background of the GxLMS as well as the results from the experiments are presented.