Single residues in the LRR domain of the wheat PM3A immune receptor can control the strength and the spectrum of the immune response.

The development of improved plant nucleotide-binding, leucine-rich repeat (LRR) immune receptors (NLRs) has mostly been based on random mutagenesis or on structural information available for specific receptors complexed with the recognized pathogen effector. Here, we use a targeted mutagenesis approach based on the natural diversity of the Pm3 powdery mildew resistance alleles present in different wheat (Triticum aestivum) genotypes. In order to understand the functional importance of the amino acid polymorphisms between the active immune receptor PM3A and the inactive ancestral variant PM3CS, we exchanged polymorphic regions and residues in the LRR domain of PM3A with the corresponding segments of PM3CS. These novel variants were functionally tested for recognition of the corresponding AVRPM3A2/F2 avirulence protein in Nicotiana benthamiana. We identified polymorphic residues in four regions of PM3A that enhance the immune response, but also residues that reduce it or result in complete loss of function. We found that the identified critical residues in PM3A modify its activation threshold towards different protein variants of AVRPM3A2/F2 . PM3A variants with a lowered threshold gave a stronger overall response and gained an extended recognition spectrum. One of these variant proteins with a single amino acid change was stably transformed into wheat, where it conferred race-specific resistance to mildew. This is a proof of concept that improved PM3A variants with an enlarged recognition spectrum can be engineered based on natural diversity by exchanging single or multiple residues that modulate resistance function.

Mid-IR refractive index sensor for detecting proteins employing an external cavity quantum cascade laser-based Mach-Zehnder interferometer.

Novel laser light sources in the mid-infrared region enable new spectroscopy schemes beyond classical absorption spectroscopy. Herein, we introduce a refractive index sensor based on a Mach-Zehnder interferometer and an external-cavity quantum cascade laser that allows rapid acquisition of high-resolution spectra of liquid-phase samples, sensitive to relative refractive index changes down to 10-7. Dispersion spectra of three model proteins in deuterated solution were recorded at concentrations as low as 0.25 mg mL-1. Comparison with Kramers-Kronig-transformed Fourier transform infrared absorbance spectra revealed high conformance, and obtained figures of merit compare well with conventional high-end FTIR spectroscopy. Finally, we performed partial least squares-based multivariate analysis of a complex ternary protein mixture to showcase the potential of dispersion spectroscopy utilizing the developed sensor to tackle complex analytical problems. The results indicate that laser-based dispersion sensing can be successfully used for qualitative and quantitative analysis of proteins.

[Use of Virtual Reality as a Component of Acute and Chronic Pain Treatment]. / Virtual Reality als Baustein in der Behandlung akuter und chronischer Schmerzen.

Future or reality? Treating acute and chronic pain is a part of the daily routine of clinical anesthesiologists. Commonly used analgesics have unwanted side effects or may even be insufficient as in chronic pain treatment. Virtual Reality (VR) could be a promising new approach which offers noninvasive therapy options for the treatment of pain. In case of the opioid misuse the adjunctive treatment is mandatory. Various phenomena occur in VR, such as immersion, presence, embodiment and Proteus effect, which can cause a change in body awareness and behavior. Experimental and clinical studies already yielded some promising results for analgesic effects for acute and chronic pain conditions using VR simulation. Potential analgesic mechanisms include distraction, cognitive behavioral change, and distance from reality, leading to neurophysiological changes at the cortical level. The quality of the virtual environment, personalized avatars, as well as the possibility of interaction and multisensory input can increase immersion, which leads to a state of presence, and thus effective VR. VR can be used as an immersive extension or alternative to mirror therapy, especially for pain disorders such as complex regional pain syndrome (CRPS) or phantom limb pain. VR can be supplemented by gamification, which increases intrinsic motivation, well-being and adherence to therapy. In summary, VR could be an effective and realistic therapy option for acute and chronic pain in clinical and home settings in the future.

Social contact patterns can buffer costs of forgetting in the evolution of cooperation.

Analyses of the evolution of cooperation often rely on two simplifying assumptions: (i) individuals interact equally frequently with all social network members and (ii) they accurately remember each partner's past cooperation or defection. Here, we examine how more realistic, skewed patterns of contact-in which individuals interact primarily with only a subset of their network's members-influence cooperation. In addition, we test whether skewed contact patterns can counteract the decrease in cooperation caused by memory errors (i.e. forgetting). Finally, we compare two types of memory error that vary in whether forgotten interactions are replaced with random actions or with actions from previous encounters. We use evolutionary simulations of repeated prisoner's dilemma games that vary agents' contact patterns, forgetting rates and types of memory error. We find that highly skewed contact patterns foster cooperation and also buffer the detrimental effects of forgetting. The type of memory error used also influences cooperation rates. Our findings reveal previously neglected but important roles of contact pattern, type of memory error and the interaction of contact pattern and memory on cooperation. Although cognitive limitations may constrain the evolution of cooperation, social contact patterns can counteract some of these constraints.

Six-Port Based Interferometry for Precise Radar and Sensing Applications.

Microwave technology plays a more important role in modern industrial sensing applications. Pushed by the significant progress in monolithic microwave integrated circuit technology over the past decades, complex sensing systems operating in the microwave and even millimeter-wave range are available for reasonable costs combined with exquisite performance. In the context of industrial sensing, this stimulates new approaches for metrology based on microwave technology. An old measurement principle nearly forgotten over the years has recently gained more and more attention in both academia and industry: the six-port interferometer. This paper reviews the basic concept, investigates promising applications in remote, as well as contact-based sensing and compares the system with state-of-the-art metrology. The significant advantages will be discussed just as the limitations of the six-port architecture. Particular attention will be paid to impairment effects and non-ideal behavior, as well as compensation and linearization concepts. It will be shown that in application fields, like remote distance sensing, precise alignment measurements, as well as interferometrically-evaluated mechanical strain analysis, the six-port architecture delivers extraordinary measurement results combined with high measurement data update rates for reasonable system costs. This makes the six-port architecture a promising candidate for industrial metrology.

Mid-IR dispersion spectroscopy - A new avenue for liquid phase analysis.

Mid-IR dispersion spectroscopy is an attractive, novel approach to liquid phase analysis that extends the possibilities of traditional methods based on the detection of absorption via intensity attenuation. This technique detects inherent refractive index changes (phase shifts) induced by IR light interaction with absorbing matter. In contrast to classic absorption spectroscopy, it provides extended dynamic range, baseline-free detection, constant sensitivity, and inherent immunity to power fluctuation. In this paper, we provide a detailed experimental and theoretical characterization and verification of this method with special focus on broadband liquid sample analysis. For this purpose, we develop a compact benchtop dispersion spectroscopy setup based on an EC-QCL coupled to a Mach-Zehnder interferometer. Phase-locked interferometric detection enables to fully harness the advantages of the technique. By instrument operation in the quadrature point combined with balanced detection, the full immunity towards laser power fluctuations and the environmental noise can be achieved. On the example of ethanol (0.5-50% v/v) dissolved in water, it is experimentally demonstrated that changes of the refractive index function are linearly related to concentration also for strongly absorbing, highly concentrated samples beyond the validity of the Beer-Lambert law. Characterization of the sensitivity and noise behavior indicates that the optimum applicable pathlength for liquid analysis can be extended beyond the ones for absorption spectroscopy. Experimental demonstration of the advantages over classical absorption spectroscopy illuminates the potential of dispersion spectroscopy as upcoming robust and sensitive way of recording IR spectra of liquid samples.

The influence of π-π-stacking on the light-harvesting properties of perylene bisimide antennas that are covalently linked to a [60]fullerene.

A flexible organic triad consisting of two perylene bisimide antennas covalently linked to a [60]fullerene by flexible spacers has been synthesized and studied by electrochemistry, steady-state and time-resolved spectroscopy. We found that π-π-stacking of the two antenna subunits has considerable impact on the photophysics of the system and leads to a reduction of the effective light-harvesting efficiency of the assembly. This finding is important for light harvesting in flexible materials based on the dye antenna-fullerene concept.

External Cavity Quantum Cascade Laser-Based Mid-Infrared Dispersion Spectroscopy for Qualitative and Quantitative Analysis of Liquid-Phase Samples.

Acquisition of classical absorption spectra of liquids in the mid-IR range with quantum cascade lasers (QCLs) is often limited in sensitivity by noise from the laser source. Alternatively, measurement of molecular dispersion (i.e., refractive index) spectra poses an experimental approach that is immune to intensity fluctuations and further offers a direct relationship between the recorded signal and the sample concentration. In this work, we present an external cavity quantum cascade laser (EC-QCL) based Mach-Zehnder interferometer setup to determine dispersion spectra of liquid samples. We present two approaches for acquisition of refractive index spectra and compare the qualitative experimental results. Furthermore, the performance for quantitative analysis is evaluated. Finally, multivariate analysis of a spectrally complex mixture comprising three different sugars is performed. The obtained figures of merit by partial least squares (PLS) regression modelling compare well with standard absorption spectroscopy, demonstrating the potential of the introduced dispersion spectroscopic method for quantitative chemical analysis.

The AvrPm3-Pm3 effector-NLR interactions control both race-specific resistance and host-specificity of cereal mildews on wheat.

The wheat Pm3 resistance gene against the powdery mildew pathogen occurs as an allelic series encoding functionally different immune receptors which induce resistance upon recognition of isolate-specific avirulence (AVR) effectors from the pathogen. Here, we describe the identification of five effector proteins from the mildew pathogens of wheat, rye, and the wild grass Dactylis glomerata, specifically recognized by the PM3B, PM3C and PM3D receptors. Together with the earlier identified AVRPM3A2/F2, the recognized AVRs of PM3B/C, (AVRPM3B2/C2), and PM3D (AVRPM3D3) belong to a large group of proteins with low sequence homology but predicted structural similarities. AvrPm3b2/c2 and AvrPm3d3 are conserved in all tested isolates of wheat and rye mildew, and non-host infection assays demonstrate that Pm3b, Pm3c, and Pm3d are also restricting the growth of rye mildew on wheat. Furthermore, divergent AVR homologues from non-adapted rye and Dactylis mildews are recognized by PM3B, PM3C, or PM3D, demonstrating their involvement in host specificity.

Fungicide prochloraz and environmental pollutant dioxin induce the ABCG2 transporter in bovine mammary epithelial cells by the arylhydrocarbon receptor signaling pathway.

The molecular mechanisms by which environmental pollutants including 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or widely used imidazole fungicide prochloraz display their toxic effects in vertebrates are still not well understood. Using computer analysis, we recently identified nuclear aryl hydrocarbon receptor (AhR) binding sites termed "dioxin response elements" (DREs) in the 5'-untranslated region (5'-UTR) of efflux transporter ABCG2 (Accession No. EU570105) from the bovine mammary gland. As these regulatory motifs mediate regulation of target genes by AhR agonists including TCDD and prochloraz, we have systematically investigated the effect of both contaminants on functional ABCG2 transport activity in primary bovine mammary epithelial cells. TCDD or prochloraz doubled ABCG2-mediated Hoechst H33342 secretion. This effect was almost completely reversed by specific ABCG2 inhibitor Ko143. In further mechanistic studies, we showed that this induction was due to binding of activated AhR to DRE sequences in the ABCG2 5'-UTR. Receptor binding was significantly reduced by specific AhR antagonist salicyl amide. Induction of AhR by TCDD and prochloraz resulted in a time- and dose-dependent increase of ABCG2 gene expression and transporter protein levels. As ABCG2 represents the main mammary transporter for xenobiotics including drugs and toxins, exposure to prevalent AhR agonists may enhance transporter-mediated secretion of potential harmful compounds into milk. Through identification of mammary ABCG2 as a novel target gene of pesticide prochloraz and dioxin, our results may therefore help to improve the protection of breast-feeding infants and the consumer of dairy products.

Determination of functional ABCG2 activity and assessment of drug-ABCG2 interactions in dairy animals using a novel MDCKII in vitro model.

The ATP-binding cassette subfamily G member 2 (ABCG2) transporter is a member of the ATP-binding cassette (ABC) family of efflux carriers that mediates cellular extrusion of various drugs and toxins. In the mammary gland, ABCG2 is expressed at the apical membrane of alveolar epithelial cells and is induced during lactation. It is well established that ABCG2 plays the main role in active secretion of xenobiotics into milk of humans and mice. In contrast, no detailed information is as yet available about functional activity and substrate spectrum of ABCG2 in dairy animals. Therefore, we cloned full-length ABCG2 from bovine, ovine and caprine lactating mammary gland tissues using rapid amplification of complementary DNA (cDNA) ends polymerase chain reaction. The generated full-length ABCG2 cDNA constructs were stably transduced in MDCKII cells. Functional ABCG2 efflux activity was demonstrated with the Hoechst H33342 accumulation assay using the specific ABCG2 inhibitor Ko143. The established ruminant MDCKII-ABCG2 cell culture models in conjunction with the H33342 transport assay showed interaction of various drugs such as cefalexin and albendazole with bABCG2, oABCG2 or cABCG2. Moreover, the flavonoids equol and quercetin exhibited interaction with all ruminant ABCG2 clones. Altogether, our generated cell culture models allowed rapid and high-throughput screening of potential ruminant ABCG2 substrates and thus increase the understanding of carrier-associated secretion of xenobiotics into milk.

Mutual interplay of light harvesting and triplet sensitizing in a perylene bisimide antenna-fullerene dyad.

A flexible organic dyad consisting of a perylene bisimide antenna covalently linked to a [60]fullerene has been synthesized and studied by electrochemistry, steady-state spectroscopy, and time-resolved spectroscopy. We found that the energy absorbed by the perylene bisimide is transferred to the fullerene with an efficiency close to 100%. The fullerene in turn undergoes intersystem crossing followed by triplet energy transfer back to the perylene bisimide with an efficiency of at least 20%. Hence the perylene bisimide unit acts as an antenna for the fullerene, i.e., effectively extending the fullerene absorption far into the visible spectral range, while at the same time the fullerene acts as a triplet sensitizer for the perylene bisimide. This has severe consequences for the exploitation of the dye antenna-fullerene concept for light harvesting in solar cells.