Modeling Takotsubo syndrome with patient-specific induced pluripotent stem cell-derived cardiomyocytes

Takotsubo syndrome (TTS) is a severe acute heart disease presumably caused by high catecholamine levels resulting in wall motion abnormalities of the left ventricle. Despite the severe phenotype of TTS, the exact disease mechanisms remain elusive. This project aims to model TTS with the help of induced pluripotent stem cell (iPSC) derived cardiomyocytes (iPSC-CMs) on a patient-specific level. To achieve this goal, 4 TTS patients and 3 control probands were recruited. Human fibroblasts or peripheral blood mononuclear cells (PBMCs) were reprogrammed to iPSCs using integration-free methods. The resulting iPSCs exhibited typical iPSC-morphology and expressed specific pluripotency-related genes on messenger RNA (mRNA) and protein level. The cells could be readily differentiated into cells from all three different germ-layers in vitro and in vivo. IPSCs were successfully differentiated to iPSC-CMs with purity over 80 % and expressing cardiac markers like cTnT and α-actinin. To mimic a TTS event, iPSC-CMs were treated with increasing concentrations of catecholamines such as epinephrine (Epi) or isoproterenol (Iso). The used high catecholamine levels did not induce significant levels of apoptosis, therefore excluding acute cytotoxicity. To test electrophysiological outcomes in the iPSC-CMs, beating rate and regularity were analyzed by using the multi-electrode array (MEA) system. MEA measurements uncovered electrical stunning for iPSC-CMs stimulated with rising levels of catecholamines. TTS-iPSC-CMs were more vulnerable to this effect. The remaining TTS-iPSC-CMs showed a stronger frequency increase when stimulated with Iso than CTRL-iPSC-CMs. Functioning calcium (Ca2+) cycling is of utmost importance in cardiomyocytes since Ca2+ is a key player in excitation-contraction coupling and Ca2+ abnormalities are involved in many heart diseases. TTS-iPSC-CMs demonstrated differences in Ca2+ cycling compared to CTRL-iPSC-CMs on multiple levels: Under basal conditions and after Iso stimulation, times to 50% decay (T50) were shorter and rise times were longer in TTS-iPSC-CMs in comparison to CTRL-iPSC-CMs. CTRL-iPSC-CMs showed a higher spark frequency at higher Iso levels (1 μM), but overall, there was no difference in total Ca2+ leakage. To analyze TTS on the tissue level, engineered heart muscles (EHM) were generated and used as a 3D-model. When challenged with Iso, the TTS-iPSC-EHMs showed a significantly higher sensitivity compared to CTRL-iPSC-EHMs as shown by a lower half-maximal effective concentration (EC50). After 24 h Iso pre-stimulation, the CTRL-iPSC-EHMs exhibiting a stronger EC50 increase than TTS-iPSC-EHM, thereby exhibiting stronger receptor desensitization. Additionally, beating frequency changes after 24 h Iso stimulation were higher in TTS-iPSC-EHM compared to CTRL-iPSC-EHM, which is in line with the reported frequency differences in the 2D MEA measurements. Interestingly, TTS-iPSC-EHM showed a lower force development than CTRL-iPSC-EHM without catecholamine stimulation. Since adrenergic stimulation results in positive inotropic and chronotropic effects of the heart and thereby regulates lipid metabolism in cardiomyocytes, lipid droplets were quantified using Oil Red O. The TTS-iPSC-CMs exhibited higher lipid accumulation in comparison to CTRL-iPSC-CMs, suggesting a role of lipotoxicity in the disease. The selective β-adrenergic receptor (β-AR) blockers CGP (β1-AR blocker) and ICI (β2-AR blocker) were used to uncover the roles of different β-ARs in CTRL- and TTS-iPSC-CMs. In CTRL-iPSC-CMs, β2-AR blocker alone was enough to reduce an Iso-dependent frequency increase and an Iso-dependent T50 and rise time decrease, thus underpinning the role of the β2-ARs in CTRL-iPSC-CMs. In TTS-iPSC-CMs, β1-AR blockage abolished Iso-dependent T50 and rise time reduction, which was not the case for β2-AR blockers and therefore underpins an important role of β1-AR in TTS-iPSC-CMs. In conclusion, these results demonstrate that CTRL- and TTS-iPSC-CMs behave differently with respect to catecholamine sensitivity, Ca2+ handling, lipid accumulation, electrophysiology, and β-AR preferences. This behavior hints towards a genetic predisposition in TTS, lowering the threshold for a TTS attack.2021-12-0

Implementation of a model for identifying Essentially Derived Varieties in vegetatively propagated Calluna vulgaris varieties

<p>Abstract</p> <p>Background</p> <p>Variety protection is of high relevance for the horticultural community and juridical cases have become more frequent in a globalized economy due to essential derivation of varieties. This applies equally to <it>Calluna vulgaris</it>, a vegetatively propagated species from the <it>Ericaceae </it>family that belongs to the top-selling pot plants in Europe. We therefore analyzed the genetic diversity of 74 selected varieties and genotypes of <it>C. vulgaris </it>and 3 of <it>Erica </it>spp. by means of RAPD and iSSR fingerprinting using 168 mono- and polymorphisms. The same data set was utilized to generate a system to reliably identify Essentially Derived Varieties (EDVs) in <it>C. vulgaris</it>, which was adapted from a method suggested for lettuce and barley. This system was developed, validated and used for selected tests of interest in <it>C. vulgaris</it>.</p> <p>Results</p> <p>As expected following personal communications with breeders, a very small genetic diversity became evident within <it>C. vulgaris </it>when investigated using our molecular methods. Thus, a dendrogram-based assay to detect Essentially Derived Varieties in this species is not suitable, although varieties are propagated vegetatively. In contrast, the system applied in lettuce, which itself applies pairwise comparisons using appropriate reference sets, proved functional with this species.</p> <p>Conclusion</p> <p>The narrow gene pool detected in <it>C. vulgaris </it>may be the genetic basis for juridical conflicts between breeders. We successfully tested a methodology for identification of Essentially Derived Varieties in highly identical <it>C. vulgaris </it>genotypes and recommend this for future proof of essential derivation in <it>C. vulgaris </it>and other vegetatively propagated crops.</p

Grundlegende Untersuchungen zur Genetik, Züchtung und Blütenorganidentität von Calluna vulgaris (L.) Hull

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Förderung sportlicher Begabung und soziale Unterstützung an Eliteschulen des Sports in Brandenburg

Das Spannungsfeld der leistungssportlichen Talentförderung im Kindes- und Jugendalter ist an seinen Endpunkten durch die schulische Allgemeinbildung und die sportliche Spezialbildung zu markieren. Einen allgemeinbildenden Schulabschluss bei gleichzeitigem spitzensportlichen Engagement zu absolvieren, fordert den schulpflichtigen Athleten und Athletinnen dabei einen erheblichen Ressourceneinsatz ab, der ohne Unterstützungsleistungen des sozialen Umfelds kaum zu leisten ist. In diesem Zusammenhang geraten vor allem die Schule-Leistungssport-Verbundsysteme in den Blick, die mittels effektiver struktureller Arrangements bei der Bewältigung einer solchen dualen Karriere unterstützen sollen. Die vorliegende Arbeit geht der Frage nach, welche Mechanismen der sozialen Unterstützung bei der Bewältigung einer dualen Karriere im Kindes- und Jugendalter wirken und welche Rolle die Verbundsysteme in diesem Kontext einnehmen.Of the achievement-sporty talent promotion in the younger age is to be marked at its terminator points by the school general education and the sporty special formation. To complete a general-forming graduation with simultaneous pointed-sporty commitment, claims from the schoolable athletes thereby a substantial resources employment, which is hardly to be carried out without support achievements of the social surrounding field. In this context above all the school achievement sport compound systems turn out into the view, which are to support with effective structural arrangements during the accomplishment of such a binary career. This book follows to the question, which mechanisms of the social support work during the accomplishment of a binary career in the younger age and which role the compound systems in this context take

AFLP-based genetic mapping of the " bud-flowering" trait in heather (Calluna vulgaris)

Background: Calluna vulgaris is one of the most important landscaping plants produced in Germany. Its enormous economic success is due to the prolonged flower attractiveness of mutants in flower morphology, the so-called bud-bloomers. In this study, we present the first genetic linkage map of C. vulgaris in which we mapped a locus of the economically highly desired trait " flower type" .Results: The map was constructed in JoinMap 4.1. using 535 AFLP markers from a single mapping population. A large fraction (40%) of markers showed distorted segregation. To test the effect of segregation distortion on linkage estimation, these markers were sorted regarding their segregation ratio and added in groups to the data set. The plausibility of group formation was evaluated by comparison of the " two-way pseudo-testcross" and the " integrated" mapping approach. Furthermore, regression mapping was compared to the multipoint-likelihood algorithm. The majority of maps constructed by different combinations of these methods consisted of eight linkage groups corresponding to the chromosome number of C. vulgaris.Conclusions: All maps confirmed the independent inheritance of the most important horticultural traits " flower type" , " flower colour" , and " leaf colour". An AFLP marker for the most important breeding target " flower type" was identified. The presented genetic map of C. vulgaris can now serve as a basis for further molecular marker selection and map-based cloning of the candidate gene encoding the unique flower architecture of C. vulgaris bud-bloomers.BLE (Federal Office of Agriculture and Food, Germany)/511–06.01-28-1-43.038-0

Heat-stable metagenomic carbonic andydrases and their use

The present invention relates to polypeptides having carbonic anhydrase activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides

What can the activation energy tell about the energetics at grain boundaries in polycrystalline organic films?

Charge-carrier transport at the semiconductor-gate dielectric interface in organic field-effect transistors is critically dependent on the degree of disorder in the typically semi-crystalline semiconductor layer. Charge trapping can occur at the interface as well as in the current-carrying semiconductor layer itself. A detailed and systematic understanding of the role of grain boundaries between crystallites and how to avoid their potentially detrimental effects is still an important focus of research in the organic electronics community. A typical macroscopic measurement technique to extract information about the energetics of the grain boundaries is an activation energy measurement. Here, we compare detailed experiments on the energetic properties of monolayer thin films implemented in organic field-effect transistors, having controlled numbers of grain boundaries within the channel region to kinetic Monte-Carlo simulations of charge-carrier transport to elucidate the influence of grain boundaries on the extracted activation energies. Two important findings are: 1) whereas the energy at the grain boundary does not change with the number of grain boundaries in a thin film, both the measured and simulated activation energy increases with the number of grain boundaries. 2) In simulations where both energy barriers and valleys are present at the grain boundaries there is no systematic relation between the number of grain boundaries and extracted activation energies. We conclude, that a macroscopic measurement of the activation energy can serve as general quality indicator of the thin film, but does not allow microscopic conclusions about the energy landscape of the thin film

Entwicklung und Erprobung eines Agroforstsystems im ökologischen Landbau zur Energieholzgewinnung

Das Projekt „Entwicklung und Erprobung eines Agroforstsystems im ökologischen Landbau zur Energieholzgewinnung“ ist eine Kooperation der beiden bayerischen Landesanstalten für Landwirtschaft (LfL) und für Wald und Forstwirtschaft (LWF). Die Versuchsstandorte liegen in der nördlichen Münchener Ebene bei Freising und im südlichen Frankenjura in der Nähe von Kaisheim. Die zentrale Frage des Projektes ist, welche Erträge und Qualitäten die landwirtschaftlichen Feldfrüchte in einem Agroforstsystem im Vergleich zur herkömmlichen Bewirtschaftung ohne Bäume auf dem Acker liefern. Es wird eine positive Wirkung von Baumstreifen im Kurzumtrieb auf den Ertrag der dazwischen liegenden landwirtschaftlichen Kulturen erwartet. Außerdem wird untersucht, ob in einem Agroforstsystem die gesamte Biomasseerzeugung im Vergleich zur reinen landwirtschaftlichen Nutzung nachhaltig höher ist. Die möglichst effiziente Etablierung von schnellwachsenden Baumarten auf Flächen des Ökolandbaus ist eine weitere Fragestellung des Projekts. Erste Beobachtungen unterschiedlicher Begründungsverfahren und zum Leistungsvergleich verschiedener Hybrid-Pappelklone mit heimischen Baumarten werden vorgestellt

Benchmarking in a rotating annulus: a comparative experimental and numerical study of baroclinic wave dynamics

The differentially heated rotating annulus is a widely studied tabletop-size laboratory model of the general mid-latitude atmospheric circulation. The two most relevant factors of cyclogenesis, namely rotation and meridional temperature gradient are quite well captured in this simple arrangement. The radial temperature difference in the cylindrical tank and its rotation rate can be set so that the isothermal surfaces in the bulk tilt, leading to the formation of baroclinic waves. The signatures of these waves at the free water surface have been analyzed via infrared thermography in a wide range of rotation rates (keeping the radial temperature difference constant) and under different initial conditions. In parallel to the laboratory experiments, five groups of the MetStr\"om collaboration have conducted numerical simulations in the same parameter regime using different approaches and solvers, and applying different initial conditions and perturbations. The experimentally and numerically obtained baroclinic wave patterns have been evaluated and compared in terms of their dominant wave modes, spatio-temporal variance properties and drift rates. Thus certain ``benchmarks'' have been created that can later be used as test cases for atmospheric numerical model validation

Praktische Erfahrungen bei der Erfassung der räumlichen Variabilität von pH-Werten in Ackerböden durch das Online-Messverfahren Veris MSP

Der pH-Wert ist ein relevanter Parameter für die Ertragsfähigkeit, auf dessen räumliche Variation zu reagieren ist. Der hohe Bodenuntersuchungsaufwand verhindert dies oft in der Praxis. Die mobile Sensorplattform MSP der Firma Veris Technologies Inc. ermöglicht eine räumlich hoch aufgelöste pH-Kartierung. In diesem Artikel wird die Einsatzfähigkeit dieses Systems für den Ökolandbau unter Praxisbedingungen geprüft. Eine effiziente Erstellung von Boden-pH-Karten ist grundsätzlich möglich. Allerdings sind weitere Untersuchungen zur Vergleichbarkeit der Ergebnisse mit der in Deutschland üblichen Methode notwendig und technische Modifikationen erforderlich, um einen störungsfreien Einsatz unter Praxisbedingungen sicherzustellen