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Supplementary Table S1. GBM-sLCR features

Glioblastoma is a lethal brain tumor that exhibits heterogeneity and resistance to therapy. Our understanding of tumor homeostasis is limited by a lack of genetic tools to selectively identify tumor states and fate transitions. Here, we use glioblastoma subtype signatures to construct synthetic genetic tracing cassettes and investigate tumor heterogeneity at cellular and molecular levels, in vitro and in vivo. Through synthetic locus control regions, we demonstrate that proneural glioblastoma is a hardwired identity, whereas mesenchymal glioblastoma is an adaptive and metastable cell state driven by proinflammatory and differentiation cues and DNA damage, but not hypoxia. Importantly, we discovered that innate immune cells divert glioblastoma cells to a proneural-to-mesenchymal transition that confers therapeutic resistance. Our synthetic genetic tracing methodology is simple, scalable, and widely applicable to study homeostasis in development and diseases. In glioblastoma, the method causally links distinct (micro)environmental, genetic, and pharmacologic perturbations and mesenchymal commitment.Significance:Glioblastoma is heterogeneous and incurable. Here, we designed synthetic reporters to reflect the transcriptional output of tumor cell states and signaling pathways' activity. This method is generally applicable to study homeostasis in normal tissues and diseases. In glioblastoma, synthetic genetic tracing causally connects cellular and molecular heterogeneity to therapeutic responses.This article is highlighted in the In This Issue feature, p. 521

The identification of the most competent embryos for transfer to the uterus constitutes the main challenge of

This study presents an application of artificial neural network (ANN) and Bayesian network (BN) for evaluation of jamming risk of the shielded tunnel boring machines (TBMs) in adverse ground conditions such as squeezing grounds. The analysis is based on database of tunneling cases by numerical modeling to evaluate the ground convergence and possibility of machine entrapment. The results of initial numerical analysis were verified in comparison with some case studies. A dataset was established by performing additional numerical modeling of various scenarios based on variation of the most critical parameters affecting shield jamming. This includes compressive strength and deformation modulus of rock mass, tunnel radius, shield length, shield thickness, in situ stresses, depth of over-excavation, and skin friction between shield and rock. Using the dataset, an ANN was trained to predict the contact pressures from a series of ground properties and machine parameters. Furthermore, the continuous and discretized BNs were used to analyze the risk of shield jamming. The results of these two different BN methods are compared to the field observations and summarized in this paper. The developed risk models can estimate the required thrust force in both cases. The BN models can also be used in the cases with incomplete geological and geomechanical properties. Keywords: Bayesian network (BN), Artificial neural network (ANN), Shielded tunnel boring machine (TBM), Jamming risk, Numerical simulation, Squeezing ground

The identification of the most competent embryos for transfer to the uterus constitutes the main challenge of in-vitro fertilization (IVF). We established a metabolomic-based approach applying Fourier Transform Infrared spectroscopy (FTIR) on 130 samples of 3-days embryo culture supernatants from 26 embryos that implanted and 104 that failed. Examining the internal structure of the data by unsupervised multivariate analysis, it was observed that the supernatants of nonimplanted embryos contained highly heterogeneous spectral features. These features were overlapping with metabolic-implantation fingerprints, thus demonstrating that in establishing embryo-assessment models a one-class modelling involving only the samples with positive-implantation outcomes should be applied. Analysis of variance confirmed that the women´s age (>40 years) undermined the implantation of the embryos exhibiting implantation metabolomics, and also that constituted a condition triggering embryos to express nonimplantation metabolomics. We conclude that IVF-success rates can be significantly improved if FTIR spectroscopy is used as an embryo-selection criterion.

Das Kap. 5 beschreibt die Bedeutung des Grundwassers fur das Bauwesen, die geohydraulischen Bedingungen und die Grundlagen der Stromung von Wasser im Baugrund. Die Wirkung des ruhenden und des stromenden Grundwassers auf die Bodenkorner und den Baugrund werden erlautert. Dazu wird die Kapillarwirkung des Bodens dargestellt.

Fur Nachweise und Bemessungen von Bauwerken und Bauteilen in der Geotechnik ist der Eurocode 7 anzuwenden. Das Kap. 8 erlautert das Nachweiskonzept und die Normung in der Geotechnik. Dabei werden die Grundlagen der Bemessung beschrieben und es erfolgt eine Darstellung und Erlauterung der Geotechnischen Kategorien. Ebenso werden die Bemessungssituationen und die Grenzzustande in der Geotechnik dargestellt.

Das mechanische Verhalten bzw. das Festigkeits- und Formanderungsverhalten eines Bodens wird durch zahlreichen Faktoren beeinflusst. Im Kap. 7 werden hierfur die grundsatzlichen Zusammenhange beschrieben und erlautert. Neben der Betrachtung der Zusammenhange bzgl. der Scherfestigkeiten von Boden und den dazugehorigen Versuchen (z. B. Triaxialversuch) werden auch die Parameter fur die Verformungseigenschaften von Boden und den dazugehorigen Versuchen (z. B. Odometerversuch) aufgezeigt.

Die Standsicherheit und die Gebrauchstauglichkeit von Bauwerken (Grundbruch, Gelandebruch, Setzung, Gleitsicherheit u. a.) ist abhangig von der Art und der Grose der einwirkenden Krafte sowie von den bodenphysikalischen Eigenschaften des Bodens. Erfullen die Eigenschaften des Baugrunds nicht die zu stellenden Anforderungen, so kann eine Verbesserung der Eigenschaften in Erwagung gezogen werden. Das Kap. 11 beschreibt die gangigsten und in der Praxis etablierten Verfahren fur die Verbesserung der Eigenschaften des Baugrunds. Neben den verfahrenstechnischen Erlauterungen werden die Voraussetzungen fur die Anwendung der Verfahren sowie Angaben wie z. B. maximal verbesserbare Tiefen oder erreichbare Festigkeiten dargestellt. Des Weiteren wurden in Kap. 11 Verfahrensvergleiche mittels Tabellen zusammengestellt, auf deren Grundlage es moglich ist, eine grobe Vorauswahl fur mogliche Baugrundverbesserungsverfahren treffen zu konnen.

Unter Bodenmechanik wird die Lehre von den Kraften im Boden und ihren Wirkungen verstanden. Zum Fachgebiet Bodenmechanik zahlen die Lehre vom Baugrund, seinem Aufbau und seinen Eigenschaften sowie die Lehre von den Spannungsverhaltnissen, den Formanderungs- und den Festigkeitseigenschaften des Bodens.

Neben der Beschreibung der normativen Grundlagen werden in Kap. 4 sehr umfassend die Versuche (z. B. Sedimentationsanalyse, Tauchwageverfahren oder Proctorversuch) dargestellt, die notwendig sind, um Bodenkenngrosen zu bestimmen. Ebenso wird erlautert, welche Bedeutung die einzelnen Bodenkenngrosen haben.

Das Kap. 6 beschreibt grundsatzliche Zusammenhange zu Spannungszustanden im Boden und bildet die Grundlage fur alle nachfolgenden Kapitel. Es werden der Primarspannungs- oder Ausgangsspannungszustand, die effektiven und die totalen Spannungen, der Porenwasserdruck u und der Porenwasseruberdruck Δu und die Konsolidation erlautert.

Chromatin modulators with antagonistic functions on epithelial-mesenchymal interconversion regulate common pan-cancer genes., Epithelial-mesenchymal transition (EMT) is a developmental process hijacked by cancer cells to modulate proliferation, migration, and stress response. Whereas kinase signaling is believed to be an EMT driver, the molecular mechanisms underlying epithelial-mesenchymal interconversion are incompletely understood. Here, we show that the impact of chromatin regulators on EMT interconversion is broader than that of kinases. By combining pharmacological modulation of EMT, synthetic genetic tracing, and CRISPR interference screens, we uncovered a minority of kinases and several chromatin remodelers, writers, and readers governing homeostatic EMT in lung cancer cells. Loss of ARID1A, DOT1L, BRD2, and ZMYND8 had nondeterministic and sometimes opposite consequences on epithelial-mesenchymal interconversion. Together with RNAPII and AP-1, these antagonistic gatekeepers control chromatin of active enhancers, including pan-cancer-EMT signature genes enabling supraclassification of anatomically diverse tumors. Thus, our data uncover general principles underlying transcriptional control of cancer cell plasticity and offer a platform to systematically explore chromatin regulators in tumor-state–specific therapy.

Excavation of long tunnels by shielded TBMs is a safe, fast, and efficient method of tunneling that mitigates many risks related to ground conditions. However, TBM applications in difficult conditions such as squeezing ground, which often happens in a combination of weaker rock and/or high depth, may lead to problems such as shield jamming. Insufficient thrust force, which cannot overcome the shield skin friction between the machine and the ground, causes shield entrapment. For successful tunneling in such conditions, it is essential to study the pertinent ground and operational factors and their impact on ground behavior, to assess the possibility of shield entrapment and to develop possible safeguards to mitigate the related risks. In this study, the controlling parameters influencing the risks of tunneling by single shield TBMs in squeezing ground are examined. For this purpose, a parametric study was performed by using a full 3D numerical simulation to quantify the influence of various factors on machine entrapment. Sensitivity analysis of most important parameters and their impacts on entrapment risks has been conducted and the results are discussed in this paper. Furthermore, nonlinear regression analysis was performed to develop a relationship between ground and machine parameters, and the required thrust force needed to overcome contact/frictional forces for propelling machine forward. The result of the modeling was compiled in a database to observe the trends and develop relationship between different parameters and risk of machine entrapment. This allows for assessment of the applicability of shield TBM in weak formations along the tunnel. Furthermore, the results of the analysis can be used to evaluate the operational parameters such as the installation of a higher thrust force to overcome contact friction loads and to make a choice between increasing the overcut or using lubricants between shield and ground. The results of this study show the role of ground conditions and in-situ stresses, as well as machine operational parameters on convergence and shield pressures.

SummaryAltering ubiquitination by disruption of individual deubiquitinating enzymes (DUBs) has proven to affect hematopoietic stem cell (HSC) maintenance. However, comprehensive knowledge of DUB function during hematopoiesis in vivo is lacking. To accomplish this goal, we systematically inactivated DUBs in mouse hematopoietic progenitors using in vivo small hairpin RNAs (shRNAs) screens. We found that multiple DUBs may be individually required for hematopoiesis and that the ubiquitin-specific protease 15 (USP15) is particularly important for the maintenance of murine hematopoietic stem and progenitor cells in vitro and in vivo. Consistently, Usp15 knockout mice exhibited a reduced HSC pool. The defect was intrinsic to HSCs, as demonstrated by competitive repopulation assays. Importantly, USP15 is highly expressed in normal human hematopoietic cells and leukemias, and USP15 depletion in murine early progenitors and myeloid leukemia cells impaired in vitro expansion and increased genotoxic stress. Our study underscores the importance of DUBs in preserving normal hematopoiesis and uncovers USP15 as a critical DUB in safeguarding genome integrity in HSC and in leukemia cells.

Mit der Einfuhrung von Industrie 4.0 werden neue Anforderungen an industrielle Netzwerke gestellt. Eine der neuen Anforderungen ist, deterministische Kommunikation nicht nur in der Maschine, sondern auch Maschinenubergreifend zu ermoglichen. Deterministische Maschinen-Maschinen Kommunikation soll in Zukunft auch Dynamik, Robustheit und hohe Verfugbarkeit unterstutzen. Um herstellerubergreifende deterministische Kommunikation auf Ethernet basierenden Netzwerken umzusetzen, werden Standards benotigt. Die IEEE 802.1 Arbeitsgruppe fur Time-Sensitive-Networking (TSN) hat sich zur Aufgabe gestellt Bausteine fur deterministisches Ethernet fur zeitkritische Applikationen zu standardisieren, welche auch die erweiterten industriellen Anforderungen erfullen. Dieser Beitrag erlautert, wie sich die Bausteine fur klassenbasierte Streams zusammenfugen lassen um flexible, robuste und redundante deterministische Kommunikation zu ermoglichen

The insufficient penetration through the cell membranes is one of the major drawbacks of chemotherapeutics such as 5-fluorouracil (5-FU; 1). To improve the penetration, a useful strategy is the attachment of lipophilic moieties. Thus, we have synthesized a series of nucleolipid derivatives of 5-fluorouridine (5-FUrd; 2a), carrying lipophilic moieties at N(3) and/or at the 2',3'-O position, i.e., 3a, 3b, 4-7, and tested their cytostatic/cytotoxic activities towards three carcinoma cell lines (colon (HT-29), hepatocellular (HepG2), and renal (RENCA)) in comparison with 5-FU (1) and 5-FUrd (2a). After 48 h of incubation, four derivatives, 3a, 3b, 5, and 7, showed inhibitory effects on the survival of HT-29, HepG2, and RENCA cells. Additionally, to differentiate between anticancer and side-effects, we tested the cytotoxicity of the derivatives in human macrophages. Interestingly, the derivatives 4, 5, and 6 did not exhibit any effects on survival of THP-1 macrophages. Furthermore, we investigated the apoptosis induction of compound 1 and 2a, and the above-mentioned derivatives in HT-29 cells. Derivative 5 showed the highest significant (p

One of the major drawbacks of chemotherapeutics is their insufficient penetration through cell membranes due to a high hydrophobicity. Thus, we have synthesized a series of selected nucleolipid derivatives of 5-fluorouridine (5-FUrd; 2a), carrying lipophilic moieties at N(3) and/or in the 2',3'-O-position (i.e., 3a-7a and 3c), and tested their cytostatic/cytotoxic activities using HT-29 human colon carcinoma cells, in comparison with, e.g., 5-FU (1) and 5-FUrd (2a). Incorporation and intracellular localization of the substances under test were performed after conjugation with the fluorochrome Atto 425. We showed that all 5'-O-labelled Atto 425 derivatives were incorporated by the human HT-29 cells and accumulated in their cytoplasm. Moreover, after 24-h treatment of HT-29 human colon carcinoma cells, 1 or 2a (10, 20, 40, or 80 μM) revealed a significant (14-23 or 33-45%, resp.) decrease of the viability in comparison with the (negative) control. Interestingly, derivatives 3a and 3c (40 and 80 μM) led to a significant (77-95 or 89-96%, resp.) inhibition of survival of human HT29 cells, i.e., these two substances were ca. 63-72% or ca. 75%, respectively more effective than 5-FU (1; positive control). Furthermore, derivative 5a showed a significant, i.e., 30 and 86%, inhibition of the survival at 40 and 80 μM, respectively in comparison with the (negative) control. Some synthesized 5-FUrd derivatives turned out to be more effective than 5-FU (1) or 5-FUrd (2a).

Zur Untersuchung der Ausbildung des Porenwasserdrucks im Bereich der Ortsbrust von maschinell aufgefahren Tunnelbauwerken wurden instationare gekoppelte mechanisch-hydraulische Variationsberechnungen an einem dreidimensionalen numerischen Simulationsmodell eines Erddruckschilds unter Anwendung der Finite-Differenzen-Methode durchgefuhrt. Im vorliegenden Aufsatz werden nach einer kurzen Darstellung des Stands der Forschung das Simulationsmodell, die Kalibrierungsberechnung und die Ergebnisse der Parameterstudien vorgestellt. Investigations of pore pressure development at the working face during mechanised tunnelling. In order to investigate the pore pressure development at the working face during mechanised tunnelling, unsteady fluid-mechanical calculations with a three-dimensional numeric simulation model were performed. In the first part of the present work, a short review of the state of the art is given. In the second part, the computation model, the calibration calculation and the results of the parameter studies are discussed.