Mediated Happiness and Digital Well-Being

<p The king sat on one end of the scale and the other end was filled with gold until a balance was reached. The king’s worth was measured by his weight in gold. Presumably, this made him happy.p>The king sat on one end of the scale and the other end was filled with gold until a balance was reached. The king’s worth was measured by his weight in gold. Presumably, this made him happy. He said that the gold on the opposite end of the scale made him happy, but we only have his word and monarchs are famously devious. In any case, whether the king said he was happy and even truly believed that he was happy when he said he was happy is beside the point. The point is: Was he really happy

Mediated Happiness and Digital Well-Being

The king sat on one end of the scale and the other end was filled with gold until a balance was reached. The king’s worth was measured by his weight in gold. Presumably, this made him happy. He said that the gold on the opposite end of the scale made him happy, but we only have his word and monarchs are famously devious. In any case, whether the king said he was happy and even truly believed that he was happy when he said he was happy is beside the point. The point is: Was he really happy

Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018.

Over the past decade, the Nomenclature Committee on Cell Death (NCCD) has formulated guidelines for the definition and interpretation of cell death from morphological, biochemical, and functional perspectives. Since the field continues to expand and novel mechanisms that orchestrate multiple cell death pathways are unveiled, we propose an updated classification of cell death subroutines focusing on mechanistic and essential (as opposed to correlative and dispensable) aspects of the process. As we provide molecularly oriented definitions of terms including intrinsic apoptosis, extrinsic apoptosis, mitochondrial permeability transition (MPT)-driven necrosis, necroptosis, ferroptosis, pyroptosis, parthanatos, entotic cell death, NETotic cell death, lysosome-dependent cell death, autophagy-dependent cell death, immunogenic cell death, cellular senescence, and mitotic catastrophe, we discuss the utility of neologisms that refer to highly specialized instances of these processes. The mission of the NCCD is to provide a widely accepted nomenclature on cell death in support of the continued development of the field

La Influencia de los Medios de Comunicación sobre los Contenidos

Mucha gente sabe que la botella que contiene vino puede influir en el gusto y calidad del mismo, pero pocos se detienen a pensar que un medio de comunicación, el cual contiene un mensaje, posee el mismo poder de alterar el significado contenido en ese medio. Un buen vino puede estar contenido en una botella de vidrio o en un envase de plástico, pero como un conocedor de vinos dirá, su gusto – lo que podríamos llamar su significado- está considerablemente afectado. O bien, si el vino se almacena en una botella de cristal tallada y antigua, hecha con una aleación de vidrio y punta de metal, sabemos que el vino se vuelve tóxico. En resumen, el envase, ya sea una botella o un medio de comunicación, afecta el contenido que contiene, incluso si uno no reflexiona porque se presta toda la atención al contenido, al vino y no a la botella que entra en jueg

Búhos y ruiseñores: un examen del autoengaño

No presenta resumen

La Influencia de los Medios de Comunicación sobre los Contenidos

Mucha gente sabe que la botella que contiene vino puede influir en el gusto y calidad del mismo, pero pocos se detienen a pensar que un medio de comunicación, el cual contiene un mensaje, posee el mismo poder de alterar el significado contenido en ese medio. Un buen vino puede estar contenido en una botella de vidrio o en un envase de plástico, pero como un conocedor de vinos dirá, su gusto – lo que podríamos llamar su significado- está considerablemente afectado. O bien, si el vino se almacena en una botella de cristal tallada y antigua, hecha con una aleación de vidrio y punta de metal, sabemos que el vino se vuelve tóxico. En resumen, el envase, ya sea una botella o un medio de comunicación, afecta el contenido que contiene, incluso si uno no reflexiona porque se presta toda la atención al contenido, al vino y no a la botella que entra en jueg

Mediated Happiness and Digital Well-Being

The king sat on one end of the scale and the other end was filled with gold until a balance was reached. The king’s worth was measured by his weight in gold. Presumably, this made him happy. He said that the gold on the opposite end of the scale made him happy, but we only have his word and monarchs are famously devious. In any case, whether the king said he was happy and even truly believed that he was happy when he said he was happy is beside the point. The point is: Was he really happy

Developing tools to optimize function analysis of tyrosine phosphorylated proteins based on cell-free protein synthesis

Die Regulation zellulärer Vorgänge wird maßgeblich durch die Ausbildung von Proteininter-aktionsnetzwerken gesteuert. Während deren Ausbildung von vielen Faktoren beeinflusst wird, besteht deren Grundeinheit in der bimolekularen Interaktion zweier Proteine. In eukaryotischen Zellen ist dabei besonders die Bindung tyrosinphosphorylierter Proteine durch SH2-Domänen von fundamentaler Bedeutung (für die Signaltransduktion). Die direkte Untersuchung solcher Wechselwirkungen bildet die Grundlage für das detaillierte Verständnis zellulärer Vorgänge. Von besonderem Interesse ist dabei die Bestimmung der Bindungsaffinität, welche die Vorhersage von Bindungshierarchien erleichtert. Bisher war es aber sehr schwer, die Bindungsaffinität Phosphotyrosin- abhängiger Wechselwirkungen in Form der Gleichgewichts-Dissoziationskonstante (Kd) auf Proteinebene zu bestimmen. In der vorliegenden Arbeit wurde am Beispiel des menschlichen T-Zell Proteins ADAP gezeigt, dass sich die Untersuchung Phosphotyrosin-abhängiger Protein-Protein-Wechsel-wirkungen durch die Kombination zellfreier Proteinsynthese mit ortspezifischer Markierung verbessern und beschleunigen lässt. Dabei wurde demonstriert, dass sich eine den Erfordernissen gerechte Auswahl an biotinylierten SH2-Domänen durch zellfreie Proteinsynthese schnell und funktional darstellen lässt. Nach zellfreier Synthese, ortspezifischer Fluoreszenzmarkierung und in vitro Phosphorylierung von ADAP gelang es, durch Kombination sensitiver Pulldowns, Überstandsabreicherung und Microscale Thermophorese (MST) unbekannte Bindungen zu identifizieren, bekannte Bindungen zu bestätigen und in Form von Kd-Werten zu quantifizieren. Für die im Vorfeld vorhergesagte Interaktion zu Rasa1 konnte dabei eine bemerkenswert hohe Affinität mit einer Kd von etwa 100 nM bestimmt werden, während die Interaktion des ebenfalls als Bindungspartner vorhergesagten SH2D1A mit einer Kd von etwa 5 µM als vernachlässigbar einzustufen ist. Die Quantifizierung der Interaktionen ergab das Rahmenwerk einer Bindungshierarchie, derzufolge ADAP an Rasa1 am stärksten bindet, gefolgt von den bekannten Interaktionspartnern SLP-76 und Fyn, während SH2D1A den Grenzbereich zu unspezifischen Bindungen markiert. Zusammenfassend wird gezeigt, dass sich durch den im Rahmen dieser Arbeit etablierten Arbeitsfluss direkte Wechselwirkungen bestätigen und zu Bindungshierarchien bezüglich des zu untersuchenden Proteins erweitern lassen. Des Weiteren wurde die Eignung der Staudinger-Phosphit-Reaktion, welche die ortspezifische Generierung von Phosphoramidaten an in Proteinen eingebautes p-Azido-L-Phenylalanin ermöglicht, als Ersatz für die Phosphorylierung durch Kinasen untersucht. Die Bildung des ungeschützten Phosphoramidats wurde auf Proteinebene nachgewiesen. Eine Wechselwirkung der modifizierten Proteine mit SH2-Domänen wurde aber anhand der im Arbeitsfluss optimierten Methoden nicht beobachtet.The regulation of cellular processes is controlled by the formation of protein interaction networks. While their assembly depends on many factors, the essential event of network formation is the bimolecular interaction between two proteins. In Eukaryotic cells, the interaction between tyrosine phosphorylated proteins and SH2 domains is of major importance for signal transduction. The direct analysis of such interactions constitutes a corner stone for the detailed interpretation of cellular processes. Thereby, the determination of binding affinities is especially beneficial, as it allows the prediction of binding hierarchies. To this point, however, it was practically impossible to determine the affinity of phosphotyrosine dependent interactions on a protein level in the form of equilibrium rate dissociation constants (Kd). Using the example of the human T-cell protein ADAP, this work demonstrates that the analysis of phosphotyrosine dependent protein-protein interactions can be improved and accelerated on the basis of cell-free protein synthesis. Cell-free protein synthesis allowed the rapid synthesis of site- specifically biotinylated SH2 domains that meet the requirement of the selection. After cell-free synthesis, site-specific fluorescence labelling and in vitro phosphorylation of ADAP, the combination of sensitive pull-downs, supernatant depletion and microscale thermophoresis (MST) enabled an identification of interactions de novo, a confirmation of known interactions and their quantification in the form of Kd values. Thereby, the previously predicted interaction between ADAP and Rasa1 shows a remarkably high affinity with a Kd in the range of 100 nM, while the equally predicted interaction to SH2D1A can be rated as negligible, since a Kd in the range of only ~5 µM was measured. The affinity quantification yielded the framework of a binding hierarchy with Rasa1 as the strongest binder, followed by ADAPs known interaction partners SLP-76 and Fyn, while SH2D1A marks a threshold range towards non-specificity. In summary, the workflow established in this study allows the identification and verification of direct protein-protein interactions and the establishment of binding hierarchies. Furthermore, the Staudinger-Phosphite Reaction, which enables the site-directed formation of phosphoramidates within proteins containing p-Azido-L-phenylalanine, was tested on its applicability to replace a kinase-mediated natural tyrosine phosphorylation. The formation of the deprotected phosphoramidate could be detected. However, no interaction of the modified proteins towards SH2 domains could be observed by means of the methods optimized in the established workflow