Induced Production of 1-Methoxy-indol-3-ylmethyl Glucosinolate by Jasmonic Acid and Methyl Jasmonate in Sprouts and Leaves of Pak Choi (Brassica rapa ssp. chinensis)

Pak choi plants (Brassica rapa ssp. chinensis) were treated with different signaling molecules methyl jasmonate, jasmonic acid, linolenic acid, and methyl salicylate and were analyzed for specific changes in their glucosinolate profile. Glucosinolate levels were quantified using HPLC-DAD-UV, with focus on induction of indole glucosinolates and special emphasis on 1-methoxy-indol-3-ylmethyl glucosinolate. Furthermore, the effects of the different signaling molecules on indole glucosinolate accumulation were analyzed on the level of gene expression using semi-quantitative realtime RT-PCR of selected genes. The treatments with signaling molecules were performed on sprouts and mature leaves to determine ontogenetic differences in glucosinolate accumulation and related gene expression. The highest increase of indole glucosinolate levels, with considerable enhancement of the 1-methoxy-indol-3-ylmethyl glucosinolate content, was achieved with treatments of sprouts and mature leaves with methyl jasmonate and jasmonic acid. This increase was accompanied by increased expression of genes putatively involved in the indole glucosinolate biosynthetic pathway. The high levels of indole glucosinolates enabled the plant to preferentially produce the respective breakdown products after tissue damage. Thus, pak choi plants treated with methyl jasmonate or jasmonic acid, are a valuable tool to analyze the specific protection functions of 1-methoxy-indole-3-carbinole in the plants defense strategy in the future.     

Heterogen katalysierte Hydrierung von Kohlendioxid zu Methan unter erhöhten Drücken

Ni‐ and Ru‐containing supported catalysts were prepared and used for the CO₂ hydrogenation to methane (Sabatier reaction) in the gas phase. Tests on the effect of the reaction temperature and pressure were in the focus. ZrO₂ and γ‐Al₂O₃ were used as suitable catalyst supports. CO₂ and H₂ conversions of 70 – 80 % and selectivity to methane of > 99 % were reached. TiO₂ and SiO₂ based catalysts exclusively lead to CO and seem to be not suited for this reaction. The investigations on the pressure effect impressively demonstrated the influence on the chemical equilibrium. CO₂ and H₂ can be nearly completely converted with > 99.9 % selectivity to methane over Ru/ZrO₂.     

Epidermal 1-O-acylceramides appear with the establishment of the water permeability barrier in mice and are produced by maturating keratinocytes

1-O-Acylceramides (1-OACs) have a fatty acid esterified to the 1-hydroxyl of the sphingosine head group of the ceramide, and recently we identified these lipids as natural components of human and mouse epidermis. Here we show epidermal 1-OACs arise shortly before birth during the establishment of the water permeability barrier in mice. Fractionation of human epidermis indicates 1-OACs concentrate in the stratum corneum. During in vitro maturation into reconstructed human epidermis, human keratinocytes dramatically increase 1-OAC levels indicating they are one source of epidermal 1-OACs. In search of potential enzymes responsible for 1-OAC synthesis in vivo, we analyzed mutant mice with deficiencies of ceramide synthases (Cers2, Cers3, or Cers4), diacylglycerol acyltransferases (Dgat1 or Dgat2), elongase of very long fatty acids 3 (Elovl3), lecithin cholesterol acyltransferase (Lcat), stearoyl-CoA desaturase 1 (Scd1), or acidic ceramidase (Asah1). Overall levels of 1-OACs did not decrease in any mouse model. In Cers3 and Dgat2-deficient epidermis they even increased in correlation with deficient skin barrier function. Dagt2 deficiency reshapes 1-OAC synthesis with an increase in 1-OACs with N-linked non-hydroxylated fatty acids and a 60% decrease compared to control in levels of 1-OACs with N-linked hydroxylated palmitate. As none of the single enzyme deficiencies we examined resulted in a lack of 1-OACs, we conclude that either there is functional redundancy in forming 1-OAC and more than one enzyme is involved, and/or an unknown acyltransferase of the epidermis performs the final step of 1-OAC synthesis, the implications of which are discussed.     

Bone Mass and Osteoblast Activity Are Sex-Dependent in Mice Lacking the Estrogen Receptor α in Chondrocytes and Osteoblast Progenitor Cells

While estrogen receptor alpha (ERα) is known to be important for bone development and homeostasis, its exact function during osteoblast differentiation remains unclear. Conditional deletion of ERα during specific stages of osteoblast differentiation revealed different bone phenotypes, which were also shown to be sex-dependent. Since hypertrophic chondrocytes can transdifferentiate into osteoblasts and substantially contribute to long-bone development, we aimed to investigate the effects of ERα deletion in both osteoblast and chondrocytes on bone development and structure. Therefore, we generated mice in which the ERα gene was inactivated via a Runx2-driven cyclic recombinase (ERα^{fl/fl; Runx2Cre}). We analyzed the bones of 3-month-old ERα^{fl/fl; Runx2Cre} mice by biomechanical testing, micro-computed tomography, and cellular parameters by histology. Male ERα^{fl/fl; Runx2Cre} mice displayed a significantly increased cortical bone mass and flexural rigidity of the femurs compared to age-matched controls with no active Cre-transgene (ERα^fl/fl). By contrast, female ERα^{fl/fl; Runx2Cre} mice exhibited significant trabecular bone loss, whereas in cortical bone periosteal and endosteal diameters were reduced. Our results indicate that the ERα in osteoblast progenitors and hypertrophic chondrocytes differentially contributes to bone mass regulation in male and female mice and improves our understanding of ERα signaling in bone cells in vivo.     

An investigation into the usability of electronic voting systems for complex elections

Many studies on electronic voting evaluate their usability in the context of simple elections. Complex elections, which take place in many European countries, also merit attention. The complexity of the voting process, as well as that of the tallying and verification of the ballots, makes usability even more crucial in this context. Complex elections, both paper-based and electronic, challenge voters and electoral officials to an unusual extent. In this work, we present two studies of an electronic voting system that is tailored to the needs of complex elections. In the first study, we evaluate the effectiveness of the ballot design with respect to motivating voters to verify their ballot. Furthermore, we identify factors that motivate voters to verify, or not to verify, their ballot. The second study also addresses the effectiveness of the ballot design in terms of verification, but this time from the electoral officials’ perspective. Last, but not least, we evaluate the usability of the implemented EasyVote prototype from both the voter and electoral official perspectives. In both studies, we were able to improve effectiveness, without impacting efficiency and satisfaction. Despite these usability improvements, it became clear that voters who trusted the electronic system were unlikely to verify their ballots. Moreover, these voters failed to detect the “fraudulent” manipulations. It is clear that well-formulated interventions are required in order to encourage verification and to improve the detection of errors or fraudulent attempts.     

Injectable Peptide Decorated Functional Nanofibrous Hollow Microspheres to Direct Stem Cell Differentiation and Tissue Regeneration

Injectable microspheres are attractive stem cell carriers for minimally invasive procedures. For tissue regeneration, the microspheres need to present the critical cues to properly direct stem cell differentiation. In natural extracellular matrix (ECM), growth factors (GFs) and collagen nanofibers provide critical chemical and physical cues. However, there have been no reported technologies that integrate synthetic nanofibers and GFs into injectable microspheres. In this study, functional nanofibrous hollow microspheres (FNF‐HMS), which can covalently bind GF‐mimicking peptides, are synthesized. Two different GF‐mimicking peptides, Transforming Growth Factor‐β1 mimicking peptide Cytomodulin (CM) and Bone Morphogenetic Protein‐2 mimicking peptide P24, are separately conjugated onto the FNF‐HMS to induce distinct differentiation pathways of rabbit bone marrow‐derived mesenchymal stem cells (BMSCs). While no existing biomaterials are reported to successfully deliver CM to induce chondrogenesis, the developed FNF‐HMS are shown to effectively present CM to BMSCs and successfully induced their chondrogenesis for ­cartilage formation in both in vitro and in vivo studies. In addition, P24 is conjugated onto the newly developed FNF‐HMS and is capable of retaining its bioactivity and inducing ectopic bone formation in nude mice. These results demonstrate that the novel FNF‐HMS can effectively deliver GF‐mimicking peptides to modulate stem cell fate and tissue regeneration.     

Electrode Work Function Engineering with Phosphonic Acid Monolayers and Molecular Acceptors: Charge Redistribution Mechanisms

The uses of self‐assembled monolayers (SAMs) of dipolar molecules or of adsorbed molecular acceptors on electrode materials are common strategies to increase their work function, thereby facilitating hole injection into an organic semiconductor deposited on top. Here it is shown that a combination of both approaches can surpass the performance of the individual ones. By combined experimental and theoretical methods it is revealed that in a three‐component system, consisting of an indium‐tin‐oxide (ITO) electrode, a carbazole‐based phosphonic acid SAM, and a molecular acceptor layer on top of the SAM, charge transfer occurs from the ITO through the SAM to the acceptor layer, resulting in an electrostatic field drop over the charge‐neutral SAM. This result is in contrast to common expectations of either p‐doping the carbazole of the SAM or charge transfer complex formation between the carbazole and the acceptor molecules. A high work function of 5.7 eV is achieved with this combined system; even higher values may be accessible by exploiting the fundamental charge redistribution mechanisms identified here with other material combinations.     

Benchmarking to the Gold Standard: Hyaluronan‐Oxime Hydrogels Recapitulate Xenograft Models with In Vitro Breast Cancer Spheroid Culture

Many 3D in vitro models induce breast cancer spheroid formation; however, this alone does not recapitulate the complex in vivo phenotype. To effectively screen therapeutics, it is urgently needed to validate in vitro cancer spheroid models against the gold standard of xenografts. A new oxime‐crosslinked hyaluronan (HA) hydrogel is designed, manipulating gelation rate and mechanical properties to grow breast cancer spheroids in 3D. This HA‐oxime breast cancer model maintains the gene expression profile most similar to that of tumor xenografts based on a pan‐cancer gene expression profile (comprising 730 genes) of three different human breast cancer subtypes compared to Matrigel or conventional 2D culture. Differences in gene expression between breast cancer cultures in HA‐oxime versus Matrigel or 2D are confirmed for 12 canonical pathways by gene set variation analysis. Importantly, drug response is dependent on the culture method. Breast cancer cells respond better to the Rac inhibitor (EHT‐1864) and the PI3K inhibitor (AZD6482) when cultured in HA‐oxime versus Matrigel. This study demonstrates the superiority of an HA‐based hydrogel as a platform for in vitro breast cancer culture of both primary, patient‐derived cells and cell lines, and provides a hydrogel culture model that closely matches that in vivo.     

Standard machine learning algorithms applied to UPLC-TOF/MS metabolic fingerprinting for the discovery of wound biomarkers in Arabidopsis thaliana

Metabolomics experiments involve the simultaneous detection of a high number of metabolites leading to large multivariate datasets and computer-based applications are required to extract relevant biological information. A high-throughput metabolic fingerprinting approach based on ultra performance liquid chromatography (UPLC) and high resolution time-of-flight (TOF) mass spectrometry (MS) was developed for the detection of wound biomarkers in the model plant Arabidopsis thaliana. High-dimensional data were generated and analysed with chemometric methods.Besides, machine learning classification algorithms constitute promising tools to decipher complex metabolic phenotypes but their application remains however scarcely reported in that research field. The present work proposes a comparative evaluation of a set of diverse machine learning schemes in the context of metabolomic data with respect to their ability to provide a deeper insight into the metabolite network involved in the wound response. Standalone classifiers, i.e. J48 (decision tree), kNN (instance-based learner), SMO (support vector machine), multilayer perceptron and RBF network (neural networks) and Naive Bayes (probabilistic method), or combinations of classification and feature selection algorithms, such as Information Gain, RELIEF-F, Correlation Feature-based Selection and SVM-based methods, are concurrently assessed and cross-validation resampling procedures are used to avoid overfitting.This study demonstrates that machine learning methods represent valuable tools for the analysis of UPLC-TOF/MS metabolomic data. In addition, remarkable performance was achieved, while the models' stability showed the robustness and the interpretability potential. The results allowed drawing attention to both temporal and spatial metabolic patterns in the context of stress signalling and highlighting relevant biomarkers not evidenced with standard data treatment.     

Magnetic multicore nanoparticles for hyperthermia--influence of particle immobilization in tumour tissue on magnetic properties

When using magnetic nanoparticles as a heating source for magnetic particle hyperthermia it is of particular interest to know if the particles are free to move in the interstitial fluid or are fixed to the tumour tissue. The immobilization state determines the relaxation behaviour of the administered particles and thus their specific heating power. To investigate this behaviour, magnetic multicore nanoparticles were injected into experimentally grown tumours in mice and magnetic heating treatment was carried out in an alternating magnetic field (H = 25 kA m(-1), f = 400 kHz). The tested particles were well suited for magnetic heating treatment as they heated a tumour of about 100 mg by about 22 K within the first 60 s. Upon sacrifice, histological tumour examination showed that the particles form spots in the tissue with a mainly homogeneous particle distribution in these spots. The magnetic ex vivo characterization of the removed tumour tissue gave clear evidence for the immobilization of the particles in the tumour tissue because the particles in the tumour showed the same magnetic behaviour as immobilized particles. Therefore, the particles are not able to rotate and a temperature increase due to Brown relaxation can be neglected. To accurately estimate the heating potential of magnetic materials, the respective environments influencing the nanoparticle mobility status have to be taken into account.