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Chromatin features are widely used for genome-scale mapping of enhancers. However, discriminating active enhancers from other cis-regulatory elements, predicting enhancer strength and identifying their target genes is challenging. Here we establish histone H2B N-terminus multisite lysine acetylation (H2BNTac) as a signature of active enhancers. H2BNTac prominently marks candidate active enhancers and a subset of promoters and discriminates them from ubiquitously active promoters. Two mechanisms underlie the distinct H2BNTac specificity: (1) unlike H3K27ac, H2BNTac is specifically catalyzed by CBP/p300; (2) H2A–H2B, but not H3–H4, are rapidly exchanged through transcription-induced nucleosome remodeling. H2BNTac-positive candidate enhancers show a high validation rate in orthogonal enhancer activity assays and a vast majority of endogenously active enhancers are marked by H2BNTac and H3K27ac. Notably, H2BNTac intensity predicts enhancer strength and outperforms current state-of-the-art models in predicting CBP/p300 target genes. These findings have broad implications for generating fine-grained enhancer maps and modeling CBP/p300-dependent gene regulation.

Herkömmliche Roboter sind aufgrund der Konfiguration rigider Gelenke und empfindlicher Getriebe anfällig für Stoßbelastungen. In diesem Beitrag wird gezeigt, dass ein Robotergelenk, basierend auf einer neuartigen Topologie mit drei aktiven und drei passiven Seilen, Stoßbelastungen bewältigen kann, die bei Bearbeitungsaufgaben wie dem Hämmern auftreten können. Conventional robots are vulnerable to high impact loads due to the configuration of rigid joints and fragile gears. This paper shows that a robotic joint based on a novel topology with three active and three passive cables can cope with impact loads as they occur in machining tasks such as hammering.

Chromatin features are widely used for genome-scale mapping of enhancers. However, discriminating active enhancers from other cis-regulatory elements, predicting enhancer strength, and identifying their target genes remains challenging. Here we establish histone H2B N-terminus multisite lysine acetylation (H2BNTac) as a genuine signature of active enhancers. H2BNTac prominently marks candidate active enhancers and their target promoters and discriminates them from ubiquitously active promoters. Two mechanisms afford the distinct H2BNTac specificity. (1) Unlike H3K27ac, H2BNTac is specifically catalyzed by CBP/p300. (2) H2A-H2B, but not H3-H4, are rapidly exchanged through transcription-induced nucleosome remodeling. H2BNTac-positive candidate enhancers show a high validation rate in orthogonal enhancer activity assays, and a vast majority of endogenously active enhancers are marked by H2BNTac and H3K27ac. Notably, H2BNTac intensity predicts enhancer strength and outperforms the current state-of-the-art models in predicting enhancer target genes. These findings have broad implications for generating fine-grained enhancer maps and modeling enhancer-dependent gene regulation.

Concurrent with autonomous robots, teleoperation gains importance in industrial applications. This includes remote intervention scenarios and human-robot-cooperation during complex or harmful tasks operated from a distance. For an efficient teleoperation, an interface achieving a good user experience and relieving workload of the operators is decisive. Whereas virtual reality allows a spatial integration, environment reconstruction is often based on point clouds in combination with pre-known object models. Challenges exist in spatial representation, through bandwidth limitations and sensor interferences. Furthermore, the potential of knowledge augmentation for workload improvement is often not fully utilized. To overcome these challenges, we present a hybrid environment reconstruction method, which is adaptive regarding the integration of different data sources as well as their derived semantic information levels. For evaluation, a user study is performed. Thereby known procedures to measure usability, immersion, user experience and operator workload serve for evaluation and comparison.

Mitochondrial Rho GTPase 1 (Miro1) protein is a well-known adaptor for mitochondrial transport and also regulates mitochondrial quality control and function. Furthermore, Miro1 was associated with mitochondrial-endoplasmic reticulum (ER) contact sites (MERCs), which are key regulators of cellular calcium homeostasis and the initiation of autophagy. Impairments of these mechanisms were linked to neurodegeneration in Parkinson’s disease (PD). We recently revealed that PD fibroblasts harboring Miro1 mutations displayed dysregulations in MERC organization and abundance, affecting mitochondrial homeostasis and clearance. We hypothesize that mutant Miro1 impairs the function of MERCs and mitochondrial dynamics, altering neuronal homeostasis and integrity in PD. PD skin fibroblasts harboring the Miro1-R272Q mutation were differentiated into patient-derived neurons. Live-cell imaging and immunocytochemistry were used to study mitophagy and the organization and function of MERCs. Markers of autophagy or mitochondrial function were assessed by western blotting. Quantification of organelle juxtapositions revealed an increased number of MERCs in patient-derived neurons. Live-cell imaging results showed alterations of mitochondrial dynamics and increased sensitivity to calcium stress, as well as reduced mitochondrial clearance. Finally, western blot analysis indicated a blockage of the autophagy flux in Miro1-mutant neurons. Miro1-mutant neurons display altered ER-mitochondrial tethering compared with control neurons. This alteration likely interferes with proper MERC function, contributing to a defective autophagic flux and cytosolic calcium handling capacity. Moreover, mutant Miro1 affects mitochondrial dynamics in neurons, which may result in disrupted mitochondrial turnover and altered mitochondrial movement.

Wie bei allen Themengebieten ist auch im E-Sport die journalistische Arbeit von immenser Bedeutung. Zeitungen und TV-Sendungen berichten über das Thema, Leitmedien schreiben via Internet zum E-Sport und Bücher entstehen. Von besonderer Wichtigkeit sind für den E-Sport- Journalismus aber Portale, die sich in den Bereichen Gaming und E-Sport bewegen. Dieses Buch zeigt die wichtige Arbeit von Journalisten im E-Sport auf und was genau alles dahintersteckt.

Increasing evidence suggests that neurodevelopmental alterations might contribute to increase the susceptibility to develop neurodegenerative diseases. We investigate the occurrence of developmental abnormalities in dopaminergic neurons in a model of Parkinson’s disease (PD). We monitor the differentiation of human patient-specific neuroepithelial stem cells (NESCs) into dopaminergic neurons. Using high-throughput image analyses and single-cell RNA sequencing, we observe that the PD-associated LRRK2-G2019S mutation alters the initial phase of neuronal differentiation by accelerating cell-cycle exit with a concomitant increase in cell death. We identify the NESC-specific core regulatory circuit and a molecular mechanism underlying the observed phenotypes. The expression of NR2F1, a key transcription factor involved in neurogenesis, decreases in LRRK2-G2019S NESCs, neurons, and midbrain organoids compared to controls. We also observe accelerated dopaminergic differentiation in vivo in NR2F1-deficient mouse embryos. This suggests a pathogenic mechanism involving the LRRK2-G2019S mutation, where the dynamics of dopaminergic differentiation are modified via NR2F1. publishedVersion

Estimating the current scene and understanding the potential maneuvers are essential capabilities of automated vehicles. Most approaches rely heavily on the correctness of maps, but neglect the possibility of outdated information. We present an approach that is able to estimate lanes without relying on any map prior. The estimation is based solely on the trajectories of other traffic participants and is thereby able to incorporate complex environments. In particular, we are able to estimate the scene in the presence of heavy traffic and occlusions. The algorithm first estimates a coarse lane-level intersection model by Markov chain Monte Carlo sampling and refines it later by aligning the lane course with the measurements using a non-linear least squares formulation. We model the lanes as 1D cubic B-splines and can achieve error rates of less than 10cm within real-time., Comment: IV 2020

The p.G2019S mutation of the leucine-rich repeat kinase 2 (LRRK2) has been identified as the most prevalent genetic cause of familial and sporadic Parkinson's disease (PD). The Cre-LoxP recombination system has been used to correct the LRRK2-G2019S mutation in patient derived human induced pluripotent stem cells (hiPSCs) in order to generate isogenic controls. However, the remaining LoxP site can influence gene expression. In this study, we report the generation of a footprint-free LRRK2-G2019S isogenic hiPS cell line edited with the CRISPR/Cas9 and piggyBac technologies. We observed that the percentage of Tyrosine Hydroxylase (TH) positive neurons with a total neurite length of >2000 μm was significantly reduced in LRRK2-G2019S dopaminergic (DA) neurons. The average branch number in LRRK2-G2019S DA neurons was also decreased. In addition, we have shown that in vitro TH positive neurons with a total neurite length of >2000 μm were positive for Serine 129 phosphorylated (S129P) alpha-Synuclein (αS) and we hypothesize that S129P-αS plays a role in the maintenance or formation of long neurites. In summary, our footprint-free LRRK2-G2019S isogenic cell lines allow standardized, genetic background independent, in vitro PD modeling and provide new insights into the role of LRRK2-G2019S and S129P-αS in the pathogenesis of PD.

Summary Research on human brain development and neurological diseases is limited by the lack of advanced experimental in vitro models that truly recapitulate the complexity of the human brain. Here, we describe a robust human brain organoid system that is highly specific to the midbrain derived from regionally patterned neuroepithelial stem cells. These human midbrain organoids contain spatially organized groups of dopaminergic neurons, which make them an attractive model for the study of Parkinson’s disease. Midbrain organoids are characterized in detail for neuronal, astroglial, and oligodendrocyte differentiation. Furthermore, we show the presence of synaptic connections and electrophysiological activity. The complexity of this model is further highlighted by the myelination of neurites. The present midbrain organoid system has the potential to be used for advanced in vitro disease modeling and therapy development., Graphical Abstract, Highlights • Generation of a human in vitro midbrain model from neural precursor cells • Midbrain organoids show neuronal, astroglial, and oligodendrocyte differentiation • Midbrain organoids contain spatially organized groups of dopaminergic neurons • Detection of synaptic connections, electrophysiological activity, and myelination, Monzel and colleagues demonstrate the derivation of human midbrain-specific brain organoids from lineage-restricted human neural precursor cells. They further show that the stem cells can self-organize into spatially patterned groups of dopaminergic neurons. Moreover, the presence of synaptic connections, myelinated of neurites, and electrophysiological activity of neurons is demonstrated.

Autophagic processes play a central role in cellular homeostasis. In pathological conditions, the flow of autophagy can be affected at multiple and distinct steps of the pathway. Current analyses tools do not deliver the required detail for dissecting pathway intermediates. The development of new tools to analyze autophagic processes qualitatively and quantitatively in a more straightforward manner is required. Defining all autophagy pathway intermediates in a high-throughput manner is technologically challenging and has not been addressed yet. Here, we overcome those requirements and limitations by the developed of stable autophagy and mitophagy reporter-iPSC and the establishment of a novel high-throughput phenotyping platform utilizing automated high-content image analysis to assess autophagy and mitophagy pathway intermediates.

The etiology of Parkinson’s disease (PD) is only partially understood despite the fact that environmental causes, risk factors, and specific gene mutations are contributors to the disease. Biallelic mutations in the PTEN-induced putative kinase 1 (PINK1) gene involved in mitochondrial homeostasis, vesicle trafficking, and autophagy, are sufficient to cause PD. By comparing PD patient-derived cells, we show differences in their energetic profile, imbalanced proliferation, apoptosis, mitophagy, and a reduced differentiation efficiency to dopaminergic neurons compared to control cells. Using CRISPR/Cas9 gene editing, correction of a patient’s point mutation ameliorated the metabolic properties and neuronal firing rates but without reversing the differentiation phenotype. However, treatment with 2-Hydroxypropyl-β-Cyclodextrin (HP-β-CD) increased the mitophagy capacity of neurons leading to an improved dopaminergic differentiation of patient specific neurons in midbrain organoids. In conclusion, we show that treatment with a repurposed compound is sufficient for restoring dopaminergic differentiation of PD patient-derived cells.

Parkinson's disease (PD) is a multifactorial disorder with complex etiology. The most prevalent PD associated mutation, LRRK2-G2019S is linked to familial and sporadic cases. Based on the multitude of genetic predispositions in PD and the incomplete penetrance of LRRK2-G2019S, we hypothesize that modifiers in the patients' genetic background act as susceptibility factors for developing PD. To assess LRRK2-G2019S modifiers, we used human induced pluripotent stem cell-derived neuroepithelial stem cells (NESCs). Isogenic controls distinguish between LRRK2-G2019S dependent and independent cellular phenotypes. LRRK2-G2019S patient and healthy mutagenized lines showed altered NESC self-renewal and viability, as well as impaired serine metabolism. In patient cells, phenotypes were only partly LRRK2-G2019S dependent, suggesting a significant contribution of the genetic background. In this context we identified the gene serine racemase (SRR) as a novel patient-specific, developmental, genetic modifier contributing to the aberrant phenotypes. Its enzymatic product, n-serine, rescued altered cellular phenotypes. Susceptibility factors in the genetic background, such as SRR, could be new targets for early PD diagnosis and treatment.

The metazoan-specific acetyltransferase p300/CBP is involved in activating signal-induced, enhancer-mediated transcription of cell-type-specific genes. However, the global kinetics and mechanisms of p300/CBP activity-dependent transcription activation remain poorly understood. We performed genome-wide, time-resolved analyses to show that enhancers and super-enhancers are dynamically activated through p300/CBP-catalyzed acetylation, deactivated by the opposing deacetylase activity, and kinetic acetylation directly contributes to maintaining cell identity at very rapid (minutes) timescales. The acetyltransferase activity is dispensable for the recruitment of p300/CBP and transcription factors but essential for promoting the recruitment of TFIID and RNAPII at virtually all enhancers and enhancer-regulated genes. This identifies pre-initiation complex assembly as a dynamically controlled step in the transcription cycle and reveals p300/CBP-catalyzed acetylation as the signal that specifically promotes transcription initiation at enhancer-regulated genes. We propose that p300/CBP activity uses a "recruit-and-release" mechanism to simultaneously promote RNAPII recruitment and pause release and thereby enables kinetic activation of enhancer-mediated transcription.

Modern cars are becoming increasingly connected. While connectivity enhances the car’s functional portfolio, it fosters the relevance of information privacy in the private vehicle. This research paper sets out to elucidate the role of data disclosure in the acceptance of connected services in the car. Based on Davis’ Technology Acceptance Model, we postulate an acceptance model which accounts for informational privacy in the connected car. In a high-fidelity driving simulator study, 116 participants interacted with a connected parking service and subsequently responded to an acceptance questionnaire. Structural equation modelling revealed a significant influence of privacy-related factors on attitude towards using the system, which in turn directly influences usage intention. The results underscore the relevance of informational privacy for the acceptance of connected vehicular services.

The advent of connected vehicles has increased the relevance of privacy in cars. While current approaches to increase security and privacy in connected vehicles are mainly driven from technological perspectives, users do not have active control over their personal data. Therefore, the user-centered privacy-aware control system PrivacyController (PRICON) has been developed which incorporates expertise from judicial, technical and user-centered perspectives. PRICON provides users with a user-friendly possibility to define self-determined privacy policies which are applied to the vehicular system. In this paper, we report the evaluation of PRICON from a legal, technical and user-centered point-of-view. The evaluation results are discussed and practical implications are derived.

The advent of connected vehicles has increased the relevance of privacy in cars. Thus, a system is required that increases transparency and helps passengers to control their privacy in a self-determined manner. Here, we derive an interface based on the privacy calculus model and prove the validity of the theory-driven design approach in course of a usability test. Based on fourteen participants, we demonstrate an above-average usability of the theory-driven design which incorporates the visual comparison of functional benefits and privacy risks. Interviews reveal that users especially acknowledge this direct comparison and underscore the successful implementation of the privacy calculus model in a vehicular privacy user interface.

Connectivity is one of the major prerequisites of automated driving. Enabled by numerous connected sensors, new cars offer new functionalities, provide higher security levels and promise to enhance the comfort of travelling. However, by connecting a vehicle with its environment, the car becomes more transparent. The integration of the car into a smart grid seems to conflict with the users’ expectation of their car as a private retreat, thus reducing the acceptance and usage adoption of connected cars. This article aims at helping developers and engineers to consider the user’s expectations when designing a connected car. Furthermore, this article reviews and compares recent international surveys on user’s privacy with our own results on the user’s attitude towards connected vehicular services.