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Crop protection strategies relying on the improvement of the natural plant immune system via genetic engineering are sustainable solutions against the pathogen thread on food security. Here we describe a novel way to improve the plant immune system by immune protease engineering. As proof of concept, we increased resistance against the late blight pathogen Phytopththora infestans by rendering the tomato secreted immune protease Pip1 insensitive to the P. infestans-secreted inhibitor Epic2B. This concept can be applied to secreted immune proteases in crops by precision breeding.

A facile multicomponent, catalyst-free oxazolidinone synthesis from primary aliphatic or aromatic amines, dibromoethane (DBE), and the usage of either cesium carbonate or cesium hydrogencarbonate as the simultaneous base and C1 source is reported. The applicability of this technically simple reaction was demonstrated by a broad scope with generally high yields, enabling concise late-stage functionalization of amino groups into N-substituted oxazolidinones. The proposed operating reaction mechanism consists of a first-step nucleophilic substitution reaction between DBE and the primary amine, followed by the formation of a carbamate or carbonate intermediate and subsequent cyclization. Additional versatility of the herein-developed protocol has been showcased in a medicinal chemistry approach by the generation of an oxazolidinone-modified dipeptidyl peptidase 8 (DPP8) inhibitor. in press

The natural product family of the fusicoccanes (FCs) has been shown to display anti-cancer activity, especially when combined with established therapeutic agents. FCs stabilize 14-3-3 protein-protein interactions (PPIs). Here, we tested combinations of a small library of FCs with interferon α (IFNα) on different cancer cell lines and report a proteomics approach to identify the specific 14-3-3 PPIs that are induced by IFNα and stabilized by FCs in OVCAR-3 cells. Among the identified 14-3-3 target proteins are THEMIS2, receptor interacting protein kinase 2 (RIPK2), EIF2AK2, and several members of the LDB1 complex. Biophysical and structural biology studies confirm these 14-3-3 PPIs as physical targets of FC stabilization, and transcriptome as well as pathway analyses suggest possible explanations for the observed synergistic effect of IFNα/FC treatment on cancer cells. This study elucidates the polypharmacological effects of FCs in cancer cells and identifies potential targets from the vast interactome of 14-3-3s for therapeutic intervention in oncology.

How the replicative Mcm2-7 helicase is activated during replication origin firing remains largely unknown. Our biochemical and structural studies reported here, demonstrate that the helicase activator GINS interacts with TopBP1 through two separate binding surfaces, the first involving a stretch of highly conserved amino acids in the TopBP1-GINI region, the second a surface located on TopBP1-BRCT4. The two surfaces bind to opposite ends of the A domain of the GINS subunit Psf1, and their cooperation is required for a biochemically stable TopBP1-GINS interaction. The GINI and BRCT4 domains also cooperate during replication origin firing, as revealed by immuno-replacement experiments inXenopusegg extracts using different sets of mutations in either interface. Furthermore, the TopBP1-GINS interaction is incompatible with simultaneous binding of DNA polymerase epsilon to GINS when bound to Mcm2-7-Cdc45. Our TopBP1-GINS model predicts the coordination of three molecular processes, DNA polymerase epsilon arrival, TopBP1 ejection and GINS integration into Mcm2-7-Cdc45.

The extracellular space of plant tissues contains hundreds of hydrolases that might harm colonising microbes. Successful pathogens may suppress these hydrolases to enable disease. Here, we report the dynamics of extracellular hydrolases in Nicotiana benthamiana upon infection with Pseudomonas syringae. Using activity-based proteomics with a cocktail of biotinylated probes, we simultaneously monitored 171 active hydrolases, including 109 serine hydrolases (SHs), 49 glycosidases (GHs) and 13 cysteine proteases (CPs). The activity of 82 of these hydrolases (mostly SHs) increases during infection, while the activity of 60 hydrolases (mostly GHs and CPs) is suppressed during infection. Active β-galactosidase-1 (BGAL1) is amongst the suppressed hydrolases, consistent with production of the BGAL1 inhibitor by P. syringae. One of the other suppressed hydrolases, the pathogenesis-related NbPR3, decreases bacterial growth when transiently overexpressed. This is dependent on its active site, revealing a role for NbPR3 activity in antibacterial immunity. Despite being annotated as a chitinase, NbPR3 does not possess chitinase activity and contains an E112Q active site substitution that is essential for antibacterial activity and is present only in Nicotiana species. This study introduces a powerful approach to reveal novel components of extracellular immunity, exemplified by the discovery of the suppression of neo-functionalised Nicotiana-specific antibacterial NbPR3. in press

The AAA-ATPase VCP/p97/Cdc48 unfolds proteins by threading them through its central pore, but how substrates are recognized and inserted into the pore in diverse pathways has remained controversial. Here, we show that p97, with its adapter p37, binds an internal recognition site (IRS) within inhibitor-3 (I3) and then threads a peptide loop into its channel to strip I3 off protein phosphatase-1 (PP1). Of note, the IRS is adjacent to the prime interaction site of I3 to PP1, and IRS mutations block I3 processing both in vitro and in cells. In contrast, amino- and carboxy-terminal regions of I3 are not required, and even circularization of I3 does not prevent I3 processing. This was confirmed by an in vitro Forster resonance energy transfer assay that allowed kinetic analysis of the reaction. Thus, our data uncover how PP1 is released from its inhibitory partner for activation and demonstrate a remarkable plasticity in substrate threading by p97.

The extracellular space of plant tissues contains hundreds of hydrolases that might harm colonizing microbes. Successful pathogens may suppress these hydrolases to enable disease. Here, we report the dynamics of extracellular hydrolases in leaves upon infection withPseudomonas syringae.Using activity-based proteomics with a cocktail of biotinylated probes we simultaneously monitored 171 active hydrolases, including 109 serine hydrolases (SHs), 49 glycosidases (GHs) and 13 cysteine proteases (CPs). The activity of 82 of these hydrolases (mostly SHs) increases during infection, whilst the activity of 60 hydrolases (mostly GHs and CPs) is suppressed during infection. Active β-galactosidase-1 (BGAL1) is amongst the suppressed hydrolases, consistent with production of the BGAL1 inhibitor byP. syringae. One of the other suppressed hydrolases, the pathogenesis-relatedNbPR3, decreases bacterial growth when transiently overexpressed. This is dependent on its active site, revealing a role forNbPR3 activity in antibacterial immunity. Despite being annotated as a chitinase,NbPR3 does not possess chitinase activity, and contains a E112Q active site substitution that is essential for antibacterial activity and is conserved only inNicotianaspecies.This study introduces a powerful approach to reveal novel components of extracellular immunity, exemplified by the discovery of the suppression of neo-functionalisedNicotiana-specificantibacterialNbPR3.

Primary attachment of rhizobia to host legume roots depends on pH and is the first physical interaction during nodulation. Genome-wide insertion sequencing, luminescence-based attachment assays and proteomic analysis demonstrate primary attachment ofRhizobium leguminosarumbiovarviciae3841 toPisum sativum(pea) roots is more complex than previously thought. In total, 115 proteins are needed for initial attachment under one or more test conditions (acid, neutral or alkaline pH), with 22 required under all conditions. These include cell-surface filamentous hemagglutinin adhesin (RL4382) and its transporter (RL4381), transmembrane protein RL2400, RL3752 (PssA, glycosyl transferase) affecting capsular polysaccharide and transcriptional regulator RL4145 (PckR). RNASeq was used to determine targets of RL4145 (PckR) and regulator RL3453. The 54 proteins required for attachment at pH 7.0 were investigated for nodulation phenotypes. Glucomannan biosynthesis protein A (GmsA) is needed at pH 6.5 and pH 7.0. Membrane proteins DgkA and ImpA are required specifically at pH 6.5, and RpoZ at pH 7.5. Sonicated cell surface fractions inhibited root attachment at alkaline pH but no overlap between proteins identified by proteomic and INseq analysis, suggests there is no single rhicadhesin needed for alkaline attachment. Our results demonstrate the complexity of primary root attachment and diversity of mechanisms involved.

Microbial communities are significant drivers of global biogeochemical cycles, yet accurate function prediction of their proteome and discerning their activityin situfor bioprospecting remains challenging. Here, we present environmental activity-based protein profiling (eABPP) as a novel proteomics-based approach bridging the gap between environmental genomics, correct function annotation andin situenzyme activity. As a showcase, we report the successful identification of active thermostable serine hydrolases by combining genome-resolved metagenomics and mass spectrometry-based eABPP of natural microbial communities from two independent hot springs in Kamchatka, Russia. eABPP does not only advance current methodological approaches by providing evidence for enzyme and microbial activityin situbut also represents an alternative approach to sequence homology-guided biocatalyst discovery from environmental ecosystems.

The ClpC1:ClpP1P2 protease is a core component of the proteostasis system in mycobacteria. To improve the efficacy of antitubercular agents targeting the Clp protease, we characterized the mechanism of the antibiotics cyclomarin A and ecumicin. Quantitative proteomics revealed that the antibiotics cause massive proteome imbalances, including upregulation of two unannotated yet conserved stress response factors, ClpC2 and ClpC3. These proteins likely protect the Clp protease from excessive amounts of misfolded proteins or from cyclomarin A, which we show to mimic damaged proteins. To overcome the Clp security system, we developed a BacPROTAC that induces degradation of ClpC1 together with its ClpC2 caretaker. The dual Clp degrader, built from linked cyclomarin A heads, was highly efficient in killing pathogenic Mycobacterium tuberculosis, with >100-fold increased potency over the parent antibiotic. Together, our data reveal Clp scavenger proteins as important proteostasis safeguards and highlight the potential of BacPROTACs as future antibiotics.

Dipeptidyl peptidases 8 and 9 (DPP8/9) have gathered interest as drug targets due to their important roles in biological processes like immunity and tumorigenesis. Elucidation of their distinct individual functions remains an ongoing task and could benefit from the availability of novel, chemically diverse and selective chemical tools. Here, we report the activity-based protein profiling (ABPP)-mediated discovery of 4-oxo-β-lactams as potent, non-substrate-like nanomolar DPP8/9 inhibitors. X-ray crystallographic structures revealed different ligand binding modes for DPP8 and DPP9, including an unprecedented targeting of an extended S2′ (eS2′) subsite in DPP8. Biological assays confirmed inhibition at both target and cellular levels. Altogether, our integrated chemical proteomics and structure-guided small molecule design approach led to novel DPP8/9 inhibitors with alternative molecular inhibition mechanisms, delivering the highest selectivity index reported to date.

In this paper, we present a model-based reinforcement learning system where the transition model is treated in a Bayesian manner. The approach naturally lends itself to exploit expert knowledge by introducing priors to impose structure on the underlying learning task. The additional information introduced to the system means that we can learn from small amounts of data, recover an interpretable model and, importantly, provide predictions with an associated uncertainty. To show the benefits of the approach, we use a challenging data set where the dynamics of the underlying system exhibit both operational phase shifts and heteroscedastic noise. Comparing our model to NFQ and BNN+LV, we show how our approach yields human-interpretable insight about the underlying dynamics while also increasing data-efficiency.

The AAA-ATPase VCP/p97 cooperates with the SEP-domain adapters p37, UBXN2A and p47 in stripping inhibitor-3 (I3) from protein phosphatase-1 (PP1) for activation. In contrast to p97-mediated degradative processes, PP1 complex disassembly is ubiquitin-independent. It is therefore unclear how selective targeting is achieved. Using biochemical reconstitution and crosslink mass spectrometry, we show here that SEP-domain adapters use a multivalent substrate recognition strategy. An N-terminal sequence element predicted to form a helix, together with the SEP-domain, binds and engages the direct target I3 in the central pore of p97 for unfolding, while its partner PP1 is held by a linker between SHP box and UBX domain locked onto the peripheral N-domain of p97. Although the I3-binding element is functional in p47, p47 in vitro requires a transplant of the PP1-binding linker from p37 for activity stressing that both sites are essential to control specificity. Of note, unfolding is then governed by an inhibitory segment in the N-terminal region of p47, suggesting a regulatory function. Together, this study reveals how p97 adapters engage a protein complex for ubiquitin-independent disassembly while ensuring selectivity for one subunit.

This article explores origins, structure and functions of repatriate organisations created by Kazakh return migrants in Kazakhstan. The article examines the factors that stimulate Kazakh repatriate...

Proteolysis is a key catalytic event in protein and thus cellular homeostasis. Despite the importance and wide implications of proteolytic processing and degradation, methods describing the degradation of folded proteins at high temporal and spatial resolution are not well established. However, this information is required to obtain a deep mechanistic understanding of proteolytic events and their consequences. Here, we describe an integrated method comprising time-resolved mass spectrometry, circular dichroism spectroscopy and bioinformatics to reveal the sequential degradation and unfolding of the model substrate annexin A1 by the human serine protease HTRA1. This workflow represents a general strategy for obtaining precise molecular insights into protease-substrate interactions that can be conveniently adapted to studying other posttranslational modifications such as phosphorylation in dynamic protein complexes.

Reaction kinetic modeling, model-based optimization and experimental validation are performed for the direct synthesis of dimethyl ether from CO2 rich synthesis gas. Among these disciplines, experimental methods and models are aligned in a stringent way of action, i.e., the same setup and models are applied throughout the whole contribution. First, a lumped reaction kinetic model from the literature is modified and parametrized to fit a vast array of 240 data points measured in a laboratory fixed bed reactor. The data were acquired using a mechanical mixture of the commercial catalysts CuO/ZnO/Al2O3 and γ-Al2O3. For this setup, a predictive model is derived and applied within dynamic model-based optimization. Here, the single-pass COx conversion serves as objective function while the operating conditions and composition of the mixed catalyst bed are the optimization variables. Finally, the optimization results obtained numerically are validated experimentally verifying the identified performance enhancement qualitatively. The remaining quantitative deviations yield valuable insights into model and methodological weaknesses or inaccuracies, closing the loop between kinetic investigations, model-based optimization and experimental validation.

The most recent work in the field of model-based design of catalytic reactors with a special focus on the multi-level reactor design (MLRD) methodology is reviewed. The key idea of this method is to track a fluid element on its way through the — not yet specified — reactor and to optimize the reaction conditions along its way by providing the necessary material and energy fluxes. On the basis of these optimal flux profiles novel reactor concepts tailored to the needs of the reaction system can be derived and analyzed. Recent advances and extensions to handle the specifics of reaction systems with different complexity are presented. The MLRD approach enables the predictive determination of the best reaction concept considering highly innovative process intensification options. Thereby, this method provides a key stone for the development of more economical and more sustainable chemical reactors and processes of the future.

Model-based design of chemical reactors via dynamic optimization can be performed using a recently developed approach called Multi-Level Reactor Design (MLRD). As a start, the MLRD method, its extensions and applications over the last ten years are reviewed compactly. In the main part of this contribution, a further extension to the model framework applied within this method allowing for the rigorous consideration of intrapellet transport processes in heterogeneously catalyzed systems is introduced. Accordingly, a system of coupled differential equations describing the mass and energy balance of both, reactor and catalyst pellet is set up. Balance equations are kept strictly distinguished from kinetic approaches, leaving the latter to be easily exchangeable. This leads to a multimodular model structure used to incorporate different models for diffusion flux, heat transport and pressure drop. The type of the extended model structure is “pseudoheterogeneous”, i.e., the domain in which chemical reactions take place is shifted from a commonly applied pseudohomogeneous mixed phase to the catalyst interior while a solid phase on reactor scale is not modelled. Applying the extended and modularized method to methanol synthesis as a case study reveals considerable concentration and temperature gradients inside the catalyst. These are neglected implicitly in pseudohomogeneous approaches which can lead to a violation of the allowed range of operating conditions for a given catalyst. The extended MLRD model framework using a pseudoheterogeneous model overcomes this drawback and allows for apparatus independent reactor design while meeting process specific constraints in both the bulk gas and the catalyst phase. Further, a systematic investigation applying 27 combinations of diffusion flux, heat transport and pressure drop models reveals that a certain performance can be reached with any combination, while the deduced reactor design differs significantly.

The AAA-ATPase VCP/p97/Cdc48 unfolds proteins by threading them through its central pore, but how substrates are recognized and inserted into the pore in diverse pathways has remained controversial. Here, we show that p97, with its adapter p37, binds an internal recognition site (IRS) within inhibitor-3 (I3) and then threads a peptide loop into its channel to strip I3 off protein phosphatase-1 (PP1). Of note, the IRS is adjacent to the prime interaction site of I3 to PP1, and IRS mutations block I3 processing both in vitro and in cells. In contrast, amino- and carboxy-terminal regions of I3 are not required, and even circularization of I3 does not prevent I3 processing. This was confirmed by an in vitro Förster resonance energy transfer assay that allowed kinetic analysis of the reaction. Thus, our data uncover how PP1 is released from its inhibitory partner for activation and demonstrate a remarkable plasticity in substrate threading by p97.

SummaryHijacking the cellular protein degradation system offers unique opportunities for drug discovery, as exemplified by proteolysis targeting chimeras (PROTACs). Despite their superior properties over classical inhibitors, it has so far not been possible to reprogram the bacterial degradation machinery to interfere with microbial infections. Here, we develop small-molecule degraders, so-called BacPROTACs, that bind to the substrate receptor of the ClpC:ClpP protease, priming neo-substrates for degradation. In addition to their targeting function, BacPROTACs activate ClpC, transforming the resting unfoldase into its functional state. The induced higher-order oligomer was visualized by cryo-EM analysis, providing a structural snapshot of activated ClpC unfolding a protein substrate. Finally, degradation assays performed in mycobacteria demonstrate in vivo activity of BacPROTACs, highlighting the potential of the technology to provide next generation antibiotics.

Behorden sind hierarchisch aufgebaute Organisationen. Anders als in vielen mittelstandischen Unternehmen erfolgt die interne und externe Kommunikation sehr stark abgestimmt, Sprachregelungen werden vereinbart, nicht jeder darf via Social Media, Vortrag oder Zeitungsinterview nach ausen kommunizieren. Dies bedeutet fur Change Communication, also die Kommunikation im Rahmen von Veranderungsprojekten, dass mit langen, teils sogar langwierigen Abstimmungsprozessen zu rechnen ist.

Um Veranderungen erfolgreich durchfuhren zu konnen, ist eine entsprechende Innovationskultur notig. Im Folgenden wird beispielhaft aufgezeigt, was der Public Sector dafur noch verinnerlichen muss.

„Change mich am Arsch“ lautet der provokante Buchtitel von Axel Koch daruber, wie Unternehmen ihre Mitarbeiter und sich selbst angeblich kaputtverandern. Aufwand und Nutzen vieler Change-Prozesse stunden in keinem Verhaltnis, mahnt der Autor aus dem Econ-Verlag in seinem an vielen Stellen polemischen Bestseller aus dem Jahr 2018. Dies ist die eine Seite der Medaille, Veranderungsprojekte moglichst gut vorauszuplanen, vor allem auch zu priorisieren und auf ihre Notwendigkeit hin vorher zu prufen.

In der bayerischen Verfassung steht seit 1973, dass Rundfunk allein in öffentlich-rechtlicher Trägerschaft betrieben wird. Seit Mitte der 1980er Jahre hat sich aber gerade im Freistaat die vielfältigste privatwirtschaftliche Radio- und Fernsehlandschaft entwickelt. In jeder größeren Stadt, in allen Regierungsbezirken gibt es lokale Sender. Auf 600 Seiten bietet der Band einen Überblick, wie sich diese Medienszene unter dem Dach der Bayerischen Landeszentrale für neue Medien (BLM) entwickelt hat – von der Vorgeschichte bis in die Gegenwart. In sechs Kapiteln und mehr als 30 Aufsätzen werden Einblicke vermittelt in die Anbieterstruktur sowie die rechtlichen, technischen und ökonomischen Grundlagen, in die Programmangebote und deren Nutzung, wie auch beispielsweise in Ansätze, die Medienkompetenz zu fördern und die Qualität dessen, was da tagaus und tagein, landauf und landab gesendet wird, vergleichend zu messen. Detailstudien bieten darüber hinaus aktuelle Befunde etwa zu Ansätzen crossmedialen Arbeitens und zur Entwicklung von Redaktionsstrukturen sowie zur Stellung von Frauen in den Redaktionen, zu Hochschulradioangeboten, zur inhaltlichen Ausgestaltung von Regionalnachrichten, zur wachsenden Bedeutung auch von Podcasts, zu Musikformaten und Moderationsformen im Wandel der Jahrzehnte und zu vielem anderen mehr. Die Vielfalt vor Ort des privaten Rundfunks in Bayern wird damit umfassend abgebildet. Since 1973, the Bavarian State Constitution requires broadcasting services licensed in Bavaria to be organised under public control. However, since the mid 1980s the most diverse commercial radio and television landscape has developed in Bavaria. There are local broadcasting stations in every major city and in every government district. On 600 pages, the anthology offers an overview of how this media scene has developed under the oversight of the Bayerische Landeszentrale für neue Medien (BLM), the regulatory authority for new media in Bavaria, over the last decades. In six chapters and more than 30 articles, historians and communication scientists discuss the broadcasting structure, the legal, technical and economic basics, the program offers and their use, as well as, for example, approaches to promote media literacy and media quality. In addition, detailed studies provide current findings, for example, on approaches to cross-media work and the development of editorial structures as well as the position of women in editorial offices, campus radio offers, the content of regional news, the growing importance of podcasts, music formats and forms of moderation over the decades and much more. Thus, the anthology comprehensively represents the diversity of the commercial broadcasting in Bavaria.

Die Grundlegen fur ein erfolgreiches Change Management gelten unabhangig von der Branche, unabhangig vom konkreten Veranderungsprojekt. Trotzdem gibt es je nach Bereich ein paar Besonderheiten, auf die geachtet werden sollte. Die Kultur im Public Sector ist eine andere als in der Energie- und wiederum in der Pharmaziebranche.

Wenn eine Behorde neue Multispace-Arbeitsplatze einfuhrt, wird sofort offensichtlich, dass dies von der Behordenleitung angeordnet oder zumindest gutgeheisen werden muss. Das heist, die Rolle der obersten Fuhrungsebene im Change-Prozess ist sichtbar. Doch auf allen Ebenen kommt den Fuhrungskraften eine entscheidende Rolle bei der Durchfuhrung von Veranderungen zu.

Die Einfuhrung der E-Akte, neue Arbeitsweisen wie Multispace-Arbeitsplatze, verstarkte Burgerorientierung oder neue Software-Programme: Bei all diesen Projekten muss bedacht werden, was dies jeweils fur den individuellen Verwaltungsangestellten oder Beamten bedeutet. Denn all das sind radikale Veranderungsprozesse, die gesteuert werden mussen. Wie dies gelingen kann, wird in diesem Kapitel dargestellt.

RNA-binding proteins (RBPs) play critical roles in post-transcriptional gene regulation and are known to contribute to plant immunity. To understand the responses of cellular RBPs to an immune elicitor, we applied RNA interactome capture to Arabidopsis leaves treated with flg22. Strikingly, flg22 induced a pervasive remodelling of the cellular RBPome affecting 186 proteins. Flg22-responsive RBPs included classical RBPs involved in RNA metabolism as well as non-canonical RBPs. RBP responders detected after 2h of treatment are enriched in putative sites for post-translational modifications, which may play a regulatory role. By contrast, changes in RBP abundance becomes increasingly important for the RBPome responses to flg22 after 12h. Plant resistance to Pseudomonas syringae is strongly altered in mutant lines lacking individual flg22-responsive RBPs, supporting the importance of RBP dynamics in plant immunity. This study provides a comprehensive and systematic census of flg22 responsive plant RBPs, discovering novel components of plant immunity.

Melanoma is a highly plastic tumor characterized by dynamic interconversion of different cell identities depending on the biological context. For example, melanoma cells with high expression of the H3K4 demethylase KDM5B (JARID1B) rest in a slow-cycling, yet reversible persister state. Over time, KDM5Bhigh cells can promote rapid tumor repopulation with equilibrated KDM5B expression heterogeneity. The cellular identity of KDM5Bhigh persister cells has not been studied so far, missing an important cell state-directed treatment opportunity in melanoma. Here, we have established a doxycycline-titratable system for genetic induction of permanent intratumor expression of KDM5B and screened for chemical agents that phenocopy this effect. Transcriptional profiling and cell functional assays confirmed that the dihydropyridine phenoxyethyl 4-(2-fluorophenyl)-2,7,7-trimethyl-5-oxo-1,4,5,6,7,8-hexa-hydro-quinoline-3-carboxylate (termed Cpd1) supports high KDM5B expression and directs melanoma cells towards differentiation along the melanocytic lineage and to cell cycle-arrest. The high KDM5B state additionally prevents cell proliferation through negative regulation of cytokinetic abscission. Moreover, treatment with Cpd1 promoted the expression of the melanocyte-specific tyrosinase gene specifically sensitizing melanoma cells for the tyrosinase-processed antifolate prodrug 3-O-(3,4,5-trimethoxybenzoyl)-(-)-epicatechin (TMECG). In summary, our study provides proof-of-concept for a new dual hit strategy in melanoma, in which persister state-directed transitioning limits tumor growth and plasticity and primes melanoma cells towards lineage-specific elimination.

Abstract Due to a high unresponsiveness to chemotherapy, biofilm formation is an important medical problem that frequently occurs during infection with many bacterial pathogens. In this study, the marine sponge-derived natural compounds 4,6-dibromo-2-(2′,4′-dibromophenoxy)phenol and 3,4,6-tribromo-2-(2′,4′-dibromophenoxy)phenol were found to exhibit broad antibacterial activity against medically relevant gram-positive and gram-negative pathogens. The compounds were not only bactericidal against both replicating and stationary phase–persistent planktonic cells of methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa; they also killed biofilm-incorporated cells of both species while not affecting biofilm structural integrity. Moreover, these compounds were active against carbapenemase-producing Enterobacter sp. This simultaneous activity of compounds against different growth forms of both gram-positive and gram-negative bacteria is rare. Genome sequencing of spontaneous resistant mutants and proteome analysis suggest that resistance is mediated by downregulation of the bacterial EIIBC phosphotransferase components scrA and mtlA in MRSA likely leading to a lower uptake of the molecules. Due to their only moderate cytotoxicity against human cell lines, phenoxyphenols provide an interesting new scaffold for development of antimicrobial agents with activity against planktonic cells, persisters and biofilm-incoporated cells of ESKAPE pathogens. Key points • Brominated phenoxyphenols kill actively replicating and biofilm-incorporated bacteria. • Phosphotransferase systems mediate uptake of brominated phenoxyphenols. • Downregulation of phosphotransferase systems mediate resistance.

To improve the transport characteristics of a catalytic fixed-bed via optimal design of catalyst pellet specifications such as the pellet diameter it is necessary to account for all physicochemical phenomena influenced by the pellet. For the approximative description of the transport phenomena on the catalyst pellet scale a new shortcut method is developed in this work. It enables a generalized and system independent treatment of the aforementioned processes and can be applied to arbitrary reaction networks and reaction kinetic models. Based on linearization and decoupling of the pellet balance equations the method yields an analytical solution. This allows for model-based design of the reactor-catalyst system via dynamic optimization at reduced computational costs as the use of complex heterogeneous models is avoided. In order to ensure accurate predictions of the method, regions with high catalyst utilization are targeted. To indicate the potential of improved bed transport characteristics, the shortcut method is applied to the reactor-catalyst system for ethylene oxide synthesis. The system is optimized in terms of reducing the pressure drop while meeting other reactor performance constraints. The pressure drop could be reduced by more than 60%. The shortcut method is validated using a rigorous stand-alone model of the catalyst pellet.

The HTRA1 gene encoding an evolutionary conserved protein quality-control factor can be epigenetically silenced or inactivated by mutation under pathologic conditions such as cancer. Recent evidence suggests that the loss of HTRA1 function causes multiple phenotypes, including the acceleration of cell growth, delayed onset of senescence, centrosome amplification, and polyploidy, suggesting an implication in the regulation of the cell cycle. To address this model, we performed a large-scale proteomics study to correlate the abundance of proteins and HTRA1 levels in various cell cycle phases using label-free-quantification mass spectrometry. These data indicate that the levels of 4723 proteins fluctuated in a cell-cycle-dependent manner, 2872 in a HTRA1-dependent manner, and 1530 in a cell-cycle- and HTRA1-dependent manner. The large number of proteins affected by the modulation of HTRA1 levels supports its general role in protein homeostasis. Moreover, the detected changes in protein abundance, in combination with pull-down data, implicate HTRA1 in various cell cycle events such as DNA replication, chromosome segregation, and cell-cycle-dependent apoptosis. These results highlight the wide implications of HTRA1 in cellular physiology.