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NetworkDynamics.jl is an easy-to-use and computationally efficient package for working with heterogeneous dynamical systems on complex networks, written in Julia, a high-level, high-performance, dynamic programming language. By combining state of the art solver algorithms from DifferentialEquations.jl with efficient data structures, NetworkDynamics.jl achieves top performance while supporting advanced features like events, algebraic constraints, time-delays, noise terms and automatic differentiation., This article may be downloaded for personal use only. Any other use requires prior permission of the author and AIP Publishing. This article appeared in Chaos 31, 063133 (2021) and may be found at https://aip.scitation.org/doi/10.1063/5.0051387

Malignant pleural mesothelioma (MPM) arises from mesothelial cells lining the pleural cavity of asbestos-exposed individuals and rapidly leads to the development of pleural effusion and death. MPM harbours loss-of-function mutations in genes likeBAP1, NF2, CDKN2A, andTP53, but isolated deletion of these genes alone in mice does not cause MPM and mouse models of the disease are sparse. Here we show that a significant proportion of human MPM harbour point mutations and copy number alterations in theKRASproto-oncogene. These mutations are likely pathogenic, since ectopic expression of mutantKRASG12Din the pleural mesothelium of conditional mice causes MPM. Murine MPM cell lines derived from these tumours carry the initiatingKRASG12Dlesions, secondaryBap1alterations, and human MPM-like gene expression profiles. Moreover, they are transplantable and actionable by KRAS inhibition. Our results indicate thatKRASmutations likely play an important and underestimated role in MPM, which warrants further exploration.

Individuals who score high in self-reported Intolerance of Uncertainty (IU) display difficulties updating threat associations to safe associations. Here we sought to determine whether individuals who score high in IU can learn and retain new safety associations if given more exposure. We recorded skin conductance response, pupil dilation and expectancy ratings during an associative threat learning task with acquisition, same-day extinction and next-day extinction phases. Participants (n = 144) were assigned to either a regular exposure (32 trials of same-day and next-day extinction) or extended exposure condition (48 trials of same-day and next-day extinction). We failed to replicate previous work showing that IU is associated with poorer safety-learning indexed via SCR, although the results were at trend and in the expected direction. We found preliminary evidence for promoted safety-retention in individuals with higher Inhibitory IU in the extended exposure condition, relative to individuals with higher Inhibitory IU in the regular exposure condition, indexed via SCR. These findings further our current understanding of the role of IU in safety-learning and -retention, informing models of IU and exposure-based treatments.

Single cell genomics enables characterization of disease specific cell states, while improvements in mass spectrometry workflows bring the clinical use of body fluid proteomics within reach. The correspondence of cell state changes in diseased organs to peripheral protein signatures is currently unknown. Here, we leverage single cell RNA-seq and proteomic analysis of large pulmonary fibrosis patient cohorts to identify disease specific changes on the cellular level and their corresponding reflection in body fluid proteomes. We discovered and validated transcriptional changes in 45 cell types across three patient cohorts that translated into distinct changes in the bronchoalveolar lavage fluid and plasma proteome. These protein signatures correlated with diagnosis, lung function, smoking and injury status. Specifically, the altered expression of a novel marker of lung health, CRTAC1, in alveolar epithelium is robustly reflected in patient plasma. Our findings have direct implications for future non-invasive prediction and monitoring of pathological cell state changes in patient organs.Abstract Figure

Single cell genomics enables characterization of disease specific cell states, while improvements in mass spectrometry bring the clinical use of body fluid proteomics within reach. The correspondence of cell state changes in diseased organs to peripheral protein signatures is currently unknown. Here, we leverage single-cell RNA-seq and proteomic analysis of large pulmonary fibrosis patient cohorts to identify disease specific changes on the cellular level and their corresponding reflection in body fluid proteomes. We discovered and validated transcriptional changes in 45 cell types across three patient cohorts that translated into distinct changes in bronchoalveolar lavage fluid and plasma proteomes. These protein signatures correlated with diagnosis, lung function, smoking and injury status. Specifically, the altered expression of a novel marker of lung health, CRTAC1, in alveolar epithelium is robustly reflected in patient plasma. Our findings have direct implications for future non-invasive prediction and monitoring of pathological cell state changes in patient organs.

Fibroblasts play an important role in lung homeostasis and disease. In lung fibrosis, fibroblasts adopt a proliferative and migratory phenotype, with increased expression of alpha-smooth muscle actin (alpha SMA) and enhanced secretion of extracellular matrix components. Comprehensive profiling of fibroblast heterogeneity is limited because of a lack of specific cell-surface markers. We have previously profiled the surface proteome of primary human lung fibroblasts. Here, we sought to define and quantify a panel of cluster of differentiation (CD) markers in primary human lung fibroblasts and idiopathic pulmonary fibrosis (IPF) lung tissue, using immunofluorescence and FACS analysis. Fibroblast function was assessed by analysis of replicative senescence. We observed the presence of distinct fibroblast phenotypes in vivo, characterized by various combinations of Desmin, alpha SMA, CD36, or CD97 expression. Most markers demonstrated stable expression over passages in vitro, but significant changes were observed for CD36, CD54, CD82, CD106, and CD140a. Replicative senescence of fibroblasts was observed from passage 10 onward. CD36- and CD97-positive but alpha SMA-negative cells were present in remodeled areas of IPF lungs. Transforming growth factor (TGF)-beta treatment induced alpha SMA and collagen I expression but repressed CD36 and CD97 expression. We identified a panel of stable surface markers in human lung fibroblasts, applicable for positive-cell isolation directly from lung tissue. TGF-beta exposure represses CD36 and CD97 expression, despite increasing alpha SMA expression; we therefore identified complex surface protein changes during fibroblast-myofibroblast activation. Coexistence of quiescence and activated fibroblast subtypes in the IPF lung suggests dynamic remodeling of fibroblast activation upon subtle changes to growth factor exposure in local microenvironmental niches.

Background Idiopathic pulmonary fibrosis (IPF) is a fatal interstitial lung disease. Repetitive injury and reprogramming of the lung epithelium are thought to be critical drivers of disease progression, contributing to fibroblast activation, extracellular matrix remodeling, and subsequently loss of lung architecture and function. To date, Pirfenidone and Nintedanib are the only approved drugs known to decelerate disease progression, however, if and how these drugs affect lung epithelial cell function, remains largely unexplored. Methods We treated murine and human 3D ex vivo lung tissue cultures (3D-LTCs; generated from precision cut lung slices (PCLS)) as well as primary murine alveolar epithelial type II (pmATII) cells with Pirfenidone or Nintedanib. Murine 3D-LTCs or pmATII cells were derived from the bleomycin model of fibrosis. Early fibrotic changes were induced in human 3D-LTCs by a mixture of profibrotic factors. Epithelial and mesenchymal cell function was determined by qPCR, Western blotting, Immunofluorescent staining, and ELISA. Results Low μM concentrations of Nintedanib (1 μM) and mM concentrations of Pirfenidone (2.5 mM) reduced fibrotic gene expression including Collagen 1a1 and Fibronectin in murine and human 3D-LTCs as well as pmATII cells. Notably, Nintedanib stabilized expression of distal lung epithelial cell markers, especially Surfactant Protein C in pmATII cells as well as in murine and human 3D-LTCs. Conclusions Pirfenidone and Nintedanib exhibit distinct effects on murine and human epithelial cells, which might contribute to their anti-fibrotic action. Human 3D-LTCs represent a valuable tool to assess anti-fibrotic mechanisms of potential drugs for the treatment of IPF patients. Electronic supplementary material The online version of this article (10.1186/s12931-018-0876-y) contains supplementary material, which is available to authorized users.

Witherite [BaCO3] and norsethite [BaMg(CO3)2] are perceived as chemical and structural analogs of aragonite [CaCO3] and dolomite [CaMg(CO3)2], respectively. However, norsethite, unlike dolomite, readily precipitates from aqueous solutions at ambient conditions. This is of special interest as the dehydration barrier of Mg2+ may be a likely cause of the dolomite growth inhibition. The easiness of norsethite growth shows that the problem of dolomite formation is more complex. In order to attain a comprehensive understanding of the analog BaCO3-MgCO3 system and of the formation of ordered anhydrous Mg-bearing double carbonates, we investigated the fate and behavior of aqueous magnesium during growth of witherite. Growth experiments were conducted on witherite seeds in mixed-flow reactors at 50 °C and various Mg-concentrations (0.25-2 mM Ba2+, 0-20 mM Mg2+, pH 7.8-8.5, ionic strength 0.1 M). At Mg:Ba ratios in solution smaller than 6:1, Mg2+ did not affect witherite growth kinetics. No significant amount of Mg2+ was incorporated. The rate constant k and reaction order n for witherite growth were determined for the first time (k = 0.65 ± 0.05·10-7 mol m-2 s-1; n =1.3 ± 0.1; supersaturation Ω = "IAP" /"K" _"s" = 1-4, where IAP is the ionic activity product and Ks the solubility constant). The insensitivity of witherite growth kinetics to these levels of Mg is analogous to aragonite growth. The general absence of the formation of solid solutions in the entire BaCO3-MgCO3 system, however, is not shared by the CaCO3-MgCO3 system, for which it is well known that substitution in the 6-fold coordinated cation sites occurs extensively. Mg:Ba ratios in solution larger than 12:1 led to a replacement of witherite by norsethite. This replacement also is in strong contrast to the CaCO3-MgCO3 system, where higher temperatures and/or much longer timescales are necessary to obtain dolomite. The replacement rate of witherite at 50 °C was estimated to be ~200 times faster than the analogous replacement of aragonite by dolomite observed over 7 years at even 60 °C (Usdowski 1989). We speculate that the preferential formation of ordered norsethite over a solid solution is facilitated by the large difference in Mg2+ and Ba2+ ionic radii. Due to the presumably very high free energy of formation of the solid solution, ordering into distinct Ba- and Mg-layers is the only way to combine both cations within one phase. In the CaCO3-MgCO3 system, solid solution occurrence is common and effectively contributes to the inhibition of the formation of the ordered double carbonate dolomite over a wide range of conditions (cf. Arvidson and Mackenzie 1999).

The easiness of norsethite [BaMg(CO3)2] precipitation at ambient conditions is in vast contrast to the growth problems of other anhydrous Mg containing carbonate minerals such as magnesite [MgCO3] and dolomite [CaMg(CO3)2]. The striking structural similarity of dolomite and norsethite makes the different growth behavior even more noteworthy. In order to investigate this remarkable discrepancy, this study focuses on the fate and behavior of aqueous barium during growth of magnesite as magnesium endmember in the BaCO3-MgCO3 system. Knowledge of barium partitioning between carbonate minerals and solution, moreover, has important implications for the assessment of the mobility of toxic elements such as radium. Growth experiments have been conducted on magnesite seeds in hydrothermal mixed-flow reactors (T = 100 °C, pH ~ 7.8, 0–100 μM Ba2+, supersaturations Ω with respect to magnesite: ~ 100–200) and by hydrothermal atomic force microscopy (T = 100 °C, pH ~ 8.2, 0–50 μM Ba2+, Ω ~ 60–90). The experiments showed that aqueous barium leaves magnesite growth rates unaffected but leads to norsethite precipitation. At the conditions of the experiments, norsethite growth rates were found to be controlled by the aqueous Ba2+ concentration. Given enough Ba2+, Mg2+ withdrawal from solution by norsethite clearly exceeded the withdrawal by magnesite growth. The logarithm of the solubility product of norsethite was found to be lower than − 18.2 at 100 °C. Microscopic investigations of the growth on the (104) surface of magnesite did not reveal any signs of Ba2+ incorporation yielding a partitioning coefficient of barium between magnesite and solution in the range of 10−2 or smaller. Preferential formation of the ordered phase norsethite over a solid solution is presumably facilitated by the large difference in Mg2+ and Ba2+ ionic radii. Due to the likely very high free energy of formation of the solid solution, ordering into distinct Ba- and Mg-layers is the only way to combine both cations within one phase. The lack of a solid solution, therefore, is one important prerequisite for the direct and unimpeded growth of an ordered double carbonate from additive free solution at ambient conditions. This behavior is in great contrast to the CaCO3-MgCO3 system where solid solution occurrence is common and growth of the ordered double carbonate (i.e. dolomite) is largely inhibited below 120 °C.

Blockade of lymphotoxin beta-receptor (LT beta R) signalling restores WNT signalling and epithelial repair in a model of chronic obstructive pulmonary disease. Lymphotoxin beta-receptor (LT beta R) signalling promotes lymphoid neogenesis and the development of tertiary lymphoid structures(1,2), which are associated with severe chronic inflammatory diseases that span several organ systems(3-6). How LT beta R signalling drives chronic tissue damage particularly in the lung, the mechanism(s) that regulate this process, and whether LT beta R blockade might be of therapeutic value have remained unclear. Here we demonstrate increased expression of LT beta R ligands in adaptive and innate immune cells, enhanced non-canonical NF-kappa B signalling, and enriched LT beta R target gene expression in lung epithelial cells from patients with smoking-associated chronic obstructive pulmonary disease (COPD) and from mice chronically exposed to cigarette smoke. Therapeutic inhibition of LT beta R signalling in young and aged mice disrupted smoking-related inducible bronchus-associated lymphoid tissue, induced regeneration of lung tissue, and reverted airway fibrosis and systemic muscle wasting. Mechanistically, blockade of LT beta R signalling dampened epithelial non-canonical activation of NF-kappa B, reduced TGF beta signalling in airways, and induced regeneration by preventing epithelial cell death and activating WNT/beta-catenin signalling in alveolar epithelial progenitor cells. These findings suggest that inhibition of LT beta R signalling represents a viable therapeutic option that combines prevention of tertiary lymphoid structures(1) and inhibition of apoptosis with tissue-regenerative strategies.

Air pollution is a major risk factor for various respiratory diseases, in particular for disease progression in COPD. Therefore, it is very important to understand pathophysiological mechanisms that determine specific interactions between air pollutants and the bronchial epithelium, thereby informing future diagnostic and treatment regimen. Primary human bronchial epithelial cells (pHBECS) from healthy/ diseased lung tissue were exposed on air-liquid interface (ALI) with relevant aerosolized nanoparticles (NPs; Carbon black nanoparticles /CNP, Zinc oxide /ZnO) and LPS, using the ALICE-CLOUD system. The functional cell viability, inflammatory and anti-oxidant response were analyzed by TEER, WST-1, LDH, qPCR and ELISA. For further validation, a novel ex-vivo airway model was established by culturing native bronchial mucosa on ALI and analyzed by 3D confocal immunofluorescence. ZnO NPs (31,8µg/cm2) induced epithelial barrier impairment after 4h (81%) and reduced viability (37% after 4h and 23% after 24h). Only ZnO NPs revealed an antioxidant response with 100-fold increased HMOX-RNA expression after 24h. qPCR analysis suggested increased TNF (3-fold) and IL8-RNA (2-fold) expression, 24h after ZnO (31,8µg/cm2) exposure. The ELISA analysis suggested an 1,5-fold increased IL8 concentration 4h after ZnO (9,3ng/ml) or LPS (10,8ng/ml) exposure. ZnO but not CNP cause epithelial barrier and cell viability impairment with the involvement of inflammatory and antioxidant pathways. The novel ex-vivo culture model based on bronchial tissue might be an important tool to validate the differentiation of the pHBECs and to better understand the inflammatory processes at the level of main bronchi.

We study two generalizations of the basin of attraction of a stable state, to the case of stochastic dynamics, arbitrary regions, and finite-time horizons. This is done by introducing generalized committor functions and studying soujourn times. We show that the volume of the generalized basin, the basin stability, can be efficiently estimated using Monte Carlo--like techniques, making this concept amenable to the study of high-dimension stochastic systems. Finally, we illustrate in a set of examples that stochastic basins efficiently capture the realm of attraction of metastable sets, which parts of phase space go into long transients in deterministic systems, that they allow us to deal with numerical noise, and can detect the collapse of metastability in high-dimensional systems. We discuss two far-reaching generalizations of the basin of attraction of an attractor. The basin of attraction of an attractor are those states that eventually will get to the attractor. In a generic stochastic system, all regions will be left again; no attraction is permanent. To obtain the equivalent of the basin of attraction of a region we need to generalize the notion to cover finite-time horizons and finite regions. We do so by considering soujourn times, the fraction of time that a trajectory spends in a set, and by generalizing committor functions which arise in the study of hitting probabilities. In a simplified setting we show that these two notions reduce to the normal notions of the basin of attraction in the appropriate limits. We also show that the volume of these stochastic basins can be efficiently estimated for high-dimensional systems at computational cost comparable to that for deterministic systems. To fully illustrate the properties captured by the stochastic basins, we show a set of examples ranging from simple conceptual models to high-dimensional inhomogeneous oscillator chains. These show that stochastic basins efficiently capture metastable attraction, the presence of long transients, that they allow us to deal with numerical and approximation noise, and can detect the collapse of metastability with increasing noise in high-dimensional systems.

During the last few years, hyperthermic intrathoracic chemotherapy (HITOC) has been performed in several departments for thoracic surgery in Germany. The objective of this expert recommendation is to provide elementary recommendations for a standardised HITOC treatment, which are based on clinical experiences and research data.Between October and December 2018, a group of experts for thoracic surgery in five departments of thoracic surgery developed recommendations for the HITOC procedure in Germany. These experts were selected by the latest national survey for HITOC and had the most clinical experience with HITOC. All recommendations are based on clinical experience, the experts' research data and recent literature.All recommendations were evaluated by all participating departments in one consensus survey. Finally, a total of six main conclusions including a total of 17 recommendations were developed. For each recommendation, the strength of the consensus is presented in percentages. 100% agreement was established for nomenclature, technique, the chemotherapeutic agent, the perioperative management, the safety measures and the indications for HITOC. All experts recommended cisplatin as the first choice chemotherapeutic agent for HITOC. The dosage of cisplatin is specified in mg/mThese expert recommendations provide a standardised and consistent implementation of the HITOC procedure. On this basis, postoperative complications associated to HITOC should be reduced and comparison of the results should be improved.Die hypertherme intrathorakale Chemotherapie (HITOC) wird bereits seit einigen Jahren in mehreren thoraxchirurgischen Kliniken angewendet. Diese Expertenempfehlung soll nun basierend auf der klinischen Erfahrung und der vorhandenen Literatur grundlegende Empfehlungen für eine standardisierte Durchführung der HITOC liefern.Im Zeitraum von Oktober bis Dezember 2018 wurden Empfehlungen zur Anwendung der HITOC in Deutschland durch eine Gruppe von Experten aus 5 thoraxchirurgischen Kliniken erarbeitet. Die Auswahl der Experten mit den meisten Operationen mit HITOC erfolgte anhand einer deutschlandweiten HITOC-Umfrage aus dem Jahr 2017. Die Empfehlungen basieren auf den klinischen Erfahrungen sowie wissenschaftlichen Arbeiten bzw. Daten der einzelnen Kliniken und einer aktuellen Literaturrecherche zur HITOC.Die Empfehlungen wurden durch alle 5 teilnehmenden Kliniken in einer Konsensusumfrage bewertet. Hieraus ergaben sich 6 Schwerpunkte mit insgesamt 17 Empfehlungen. Bei jeder Empfehlung wurde eine Konsensstärke in Prozent berechnet. Eine 100%ige Konsensstärke wurde bei der Nomenklatur (HITOC), der technischen Durchführung, dem Chemotherapeutikum, dem perioperativen Management, den Schutzmaßnahmen und den Indikationen erreicht. Als Basis-Chemotherapeutikum für die HITOC wird von allen Experten Cisplatin empfohlen, da hier die meisten klinischen Erfahrungen und Daten vorliegen. Die Dosierung von Cisplatin wird in mg/mDiese Expertenempfehlungen zeigen eine standardisierte und einheitliche Durchführung der HITOC. Hierdurch sollen postoperative Komplikationen durch die HITOC vermindert und eine bessere Vergleichbarkeit der Ergebnisse ermöglicht werden.

Translation of novel discoveries to human disease is limited by the availability of human tissue-based models of disease. Precision-cut lung slices (PCLS) used as 3D lung tissue cultures (3D-LTCs) represent an elegant and biologically highly relevant 3D cell culture model, which highly resemble in situ tissue due to their complexity, biomechanics and molecular composition. Tissue slicing is widely applied in various animal models. 3D-LTCs derived from human PCLS can be used to analyze responses to novel drugs, which might further help to better understand the mechanisms and functional effects of drugs in human tissue. The preparation of PCLS from surgically resected lung tissue samples of patients, who experienced lung lobectomy, increases the accessibility of diseased and peritumoral tissue. Here, we describe a detailed protocol for the generation of human PCLS from surgically resected soft-elastic patient lung tissue. Agarose was introduced into the bronchoalveolar space of the resectates, thus preserving lung structure and increasing the tissue's stiffness, which is crucial for subsequent slicing. 500 µm thick slices were prepared from the tissue block with a vibratome. Biopsy punches taken from PCLS ensure comparable tissue sample sizes and further increase the amount of tissue samples. The generated lung tissue cultures can be applied in a variety of studies in human lung biology, including the pathophysiology and mechanisms of different diseases, such as fibrotic processes at its best at (sub-)cellular levels. The highest benefit of the 3D-LTC ex vivo model is its close representation of the in situ human lung in respect of 3D tissue architecture, cell type diversity and lung anatomy as well as the potential for assessment of tissue from individual patients, which is relevant to further develop novel strategies for precision medicine.

Previous studies of change blindness have suggested a distinction between detection and localisation of changes in a visual scene. Using a simple paradigm with an array of coloured squares, the present study aimed to further investigate differences in event-related potentials (ERPs) between trials in which participants could detect the presence of a colour change but not identify the location of the change (sense trials), versus those where participants could both detect and localise the change (localise trials). Individual differences in performance were controlled for by adjusting the difficulty of the task in real time. Behaviourally, reaction times for sense, blind, and false alarm trials were distinguishable when comparing across levels of participant certainty. In the EEG data, we found no significant differences in the visual awareness negativity ERP, contrary to previous findings. In the N2pc range, both awareness conditions (localise and sense) were significantly different to trials with no change detection (blind trials), suggesting that this ERP is not dependent on explicit awareness. Within the late parietal positivity range, all conditions were significantly different. These results suggest that changes can be ‘sensed’ without knowledge of the location of the changing object, and that participant certainty scores can provide valuable information about the perception of changes in change blindness.

Background: Malignant pleural mesothelioma (MPM) is an aggressive malignancy with few long-term survivors. Despite the dismal prognosis, hyperthermic intrathoracic chemoperfusion (HITHOC) was shown to improve survival in a selective group of patients. We analyzed the influence of HITHOC following pleurectomy and decortication on postoperative morbidity and overall survival for patients suffering from localized mesothelioma. Methods: From 2009 until 2013, 71 patients with localized pleural mesothelioma underwent pleurectomy and decortication followed by HITHOC with cisplatin and doxorubicin. We analyzed postoperative morbidity, age, overall survival and influence of macroscopic resection on survival. Results: Median patient age was 70 years (range, 65-73 years). Patients having the sarcomatoid subtype of mesothelioma showed a poor median survival of 9.2 months. In contrast, patients having the epithelioid subtype had a median survival of 17.9 months. Patients following macroscopic complete resection had a significantly better survival with 28.2 months compared to 13.1 months in patients with incomplete resection of the mesothelioma (P

Transforming growth factor-β (TGF-β)-induced fibroblast-to-myofibroblast differentiation contributes to remodeling in chronic obstructive pulmonary disease and idiopathic pulmonary fibrosis, but whether this impacts the ability of fibroblasts to support lung epithelial repair remains little explored. We pretreated human lung fibroblasts [primary (phFB) or MRC5 cells] with recombinant human TGF-β to induce myofibroblast differentiation, then cocultured them with adult mouse lung epithelial cell adhesion molecule-positive cells (EpCAM+) to investigate their capacity to support epithelial organoid formation in vitro. While control phFB and MRC5 lung fibroblasts supported organoid formation of mouse EpCAM+cells, TGF-β pretreatment of both phFB and MRC5 impaired organoid-supporting ability. We performed RNA sequencing of TGF-β-treated phFB, which revealed altered expression of key Wnt signaling pathway components and Wnt/β-catenin target genes, and modulated expression of secreted factors involved in mesenchymal-epithelial signaling. TGF-β profoundly skewed the transcriptional program induced by the Wnt/β-catenin activator CHIR99021. Supplementing organoid culture media recombinant hepatocyte growth factor or fibroblast growth factor 7 promoted organoid formation when using TGF-β pretreated fibroblasts. In conclusion, TGF-β-induced myofibroblast differentiation results in Wnt/β-catenin pathway skewing and impairs fibroblast ability to support epithelial repair likely through multiple mechanisms, including modulation of secreted growth factors.

A Correction to this paper has been published: https://doi.org/10.1038/s41586-020-03087-6

Wir berichten uber den Fall eines jungen Mannes mit einer Echinokokkuszyste. Die differenzialdiagnostische Einordnung von zystischen Lasionen der Lunge kann schwierig sein, zumal die zur Verfugung stehenden serologischen Testsysteme insbesondere bei pulmonaler Manifestation der Echinokokkose eine eingeschrankte Sensitivitat aufweisen. Die bronchoskopische Ultraschalldarstellung zystischer Lasionen bzw. die Analyse des Zystenpunktates kann zur weiteren diagnostischen Einordung beitragen. Allerdings ist im vorliegenden Fall eine eosinophile Pneumonie als Folge der bronchoskopischen Punktion zu diskutieren.

RATIONALE\nChronic obstructive pulmonary disease patients and in particular smokers are more susceptible to respiratory infections contributing to acute exacerbations of disease. The immunoproteasome is a specialized type of proteasome destined to improve major histocompatibility complex (MHC) class I-mediated antigen presentation for the resolution of intracellular infections.\nOBJECTIVES\nTo characterize immunoproteasome function in COPD and its regulation by cigarette smoke.\nMETHODS\nImmunoproteasome expression and activity were determined in bronchoalveolar lavage (BAL) and lungs of human donors, COPD, and IPF patients, as well as in cigarette smoke-exposed mice. Smoke-mediated alteration of immunoproteasome activity and MHC I surface expression were analysed in human blood-derived macrophages. Immunoproteasome-specific MHC I antigen presentation was evaluated in spleen and lung immune cells that had been smoke-exposed in vitro or in vivo.\nMEASUREMENTS AND MAIN RESULTS\nImmunoproteasome and MHC I mRNA expression was reduced in BAL cells of COPD patients and in isolated alveolar macrophages of COPD and IPF patients. Exposure of immune cells to cigarette smoke extract in vitro reduced immunoproteasome activity and impaired immunoproteasome-specific MHC I antigen presentation. In vivo, acute cigarette smoke exposure dynamically regulated immunoproteasome function and MHC I antigen presentation in mouse BAL cells. End-stage COPD lungs showed markedly impaired immunoproteasome activities.\nCONCLUSIONS\nWe here show for the first time that the activity of the immunoproteasome is impaired by cigarette smoke resulting in reduced MHC I antigen presentation. Regulation of immunoproteasome function by cigarette smoke may thus alter adaptive immune responses and add to prolonged infections and exacerbations in COPD and IPF.

// Arne Warth 1, # , Volker Endris 1 , Albrecht Stenzinger 1 , Roland Penzel 1 , Alexander Harms 1 , Thomas Duell 2 , Amir Abdollahi 3, + , Michael Lindner 4 , Peter Schirmacher 1 , Thomas Muley 5, # , Hendrik Dienemann 6, # , Ludger Fink 7, # , Alicia Morresi-Hauf 8 , Nicole Pfarr 1, 10, § , Wilko Weichert 1, 9, 10, +, § 1 Institute of Pathology, Heidelberg University, Heidelberg, Germany 2 Department of Pneumology and Thoracic Oncology, Asklepios Hospital, Munich-Gauting, Germany 3 Department of Radiation Oncology, Heidelberg University, Heidelberg, Germany 4 Department of Thoracic Surgery, Asklepios Hospital, Munich-Gauting, Germany 5 Translational Research Unit, Thoraxklinik at Heidelberg University, Heidelberg, Germany 6 Department of Thoracic Surgery, Thoraxklinik at Heidelberg University, Heidelberg, Germany 7 Institute of Pathology, Wetzlar, Germany 8 Institute of Pathology, Asklepios Hospital, Munich-Gauting, Germany 9 National Center for Tumor Diseases (NCT), Heidelberg, Germany 10 Institute of Pathology, Technical University (TUM), Munich, Germany # Translational Lung Research Centre Heidelberg, Member of the German Center for Lung Research (DZL-TLRC) + Member of the German Cancer Consortium (DKTK) § shared senior authorship Correspondence to: Arne Warth, email: arne.warth@med.uni-heidelberg.de Wilko Weichert, email: wilko.weichert@tum.de Keywords: lung cancer, molecular evolution, driver mutations, therapy Received: November 06, 2015 Accepted: March 28, 2016 Published: April 20, 2016 ABSTRACT Background: Large scale sequencing efforts defined common molecular alterations in non-small cell lung cancer (NSCLC) and revealed potentially druggable mutations. Yet, systematic data on the changes of the respective molecular profiles under standard therapy in NSCLC are limited. Results: 14 out of 68 observed coding mutations (21%) and 6 out of 33 (18%) copy number variations (CNV) were lost or gained during therapy. Mutational and CNV changes clustered in 6/37 (16%) and 3/37 (8%) patients. Changes in clinically relevant mutations were rare but present in single cases for genes such as BRAF and PIK3CA . The type of radiochemotherapy but not the duration of therapy impacted on the frequency of mutational changes. Methods: We established a lung cancer specific next-generation sequencing panel covering ~7500 hotspots of 41 genes frequently mutated in NSCLC and performed ultradeep multigene sequencing of 37 corresponding pre- and post-therapeutic formalin fixed paraffin-embedded specimens to discover mutational changes and copy number variations under neo-adjuvant radio- (RTX) and/or chemotherapy (CTX). Conclusion: We unraveled changes in common driver gene candidates in NSCLC under neo-adjuvant therapy. Our data shed first light on the genetic changes of NSCLC under conventional therapy and might be taken into account when the relevance of sequential biopsy approaches is discussed.

Motivated by the strong effect of aqueous Mg on calcite growth rates, this study used hydrothermal atomic force microscopy (HAFM) and hydrothermal mixed-flow reactor (HMFR) experiments to explore the effect of aqueous Ca on magnesite growth kinetics at 100 °C and pH ∼7.7. Obtuse step velocities on (1 0 4) surfaces during magnesite growth were measured to be 4 ± 3 nm/s at fluid saturation states, equal to the ion activity quotient divided by the equilibrium constant for the magnesite hydrolysis reaction, of 86–117. These rates do not vary systematically with aqueous Ca concentration up to 3 × 10 − 3 mol/kg. Magnesite growth rates determined by HAFM are found to be negligibly affected by the presence of aqueous Ca at these saturation states and are largely consistent with those previously reported in aqueous Ca-free systems by Saldi et al. (2009) and Gautier et al. (2015). Similarly, magnesite growth rates measured by HMFR exhibit no systematic variation on aqueous Ca concentrations. Rates in this study, however, were extended to higher degrees of fluid supersaturation with respect to magnesite than previous studies. All measured HMFR rates can be accurately described taking account the combined effects of both the spiral growth and two dimensional nucleation/growth mechanisms. Despite the lack of a clear effect of aqueous Ca on magnesite growth rates, Raman spectroscopy confirmed the incorporation of up to 8 mol percent of Ca 2+ into the growing magnesite structure.

Nanoparticle‐based targeted drug delivery holds promise for treatment of cancers. However, most approaches fail to be translated into clinical success due to ineffective tumor targeting in vivo. Here, the delivery potential of mesoporous silica nanoparticles (MSN) functionalized with targeting ligands for epidermal growth factor receptor and C─C chemokine receptor type 2 is explored in lung tumors. The addition of active targeting ligands on MSNs enhances their uptake in vitro but fails to promote specific delivery to tumors in vivo, when administered systemically via the blood or locally to the lung into immunocompetent murine lung cancer models. Ineffective tumor targeting is due to efficient clearance of the MSNs by the phagocytic cells of the liver, spleen, and lung. These limitations, however, are successfully overcome using a novel organ‐restricted vascular delivery (ORVD) approach. ORVD in isolated and perfused mouse lungs of Kras‐mutant mice enables effective nanoparticle extravasation from the tumor vasculature into the core of solid lung tumors. In this study, ORVD promotes tumor cell‐specific uptake of nanoparticles at cellular resolution independent of their functionalization with targeting ligands. Organ‐restricted vascular delivery thus opens new avenues for optimized nanoparticles for lung cancer therapy and may have broad applications for other vascularized tumor types.

Human sample, from patients or healthy donors, are a valuable link between basic research and clinic. Especially in translational research, they play an essential role in understanding development and progression of diseases as well as in developing new diagnostic and therapeutic tools. Stored in biobanks, fast access to appropriate material becomes possible. However, biobanking in a clinical context faces several challenges. In practice, collecting samples during clinical routine does not allow to strictly adhere to protocols of sample collection in all aspects. This may influence sample quality to variable degrees. Time from sample draw to asservation is a variable factor, and influences of prolonged storage at ambient temperature of tissues are not well understood. We investigated whether delays between 5 minutes and 3 hours, and the use of RNAlater RNA-preserving reagent would lead to a relevant drop in sample quality, measured by quantitative mRNA expression analysis. Our findings suggest that even under ambient conditions, delays up to 3 hours do not have a major impact on sample quality as long as the tissue remains intact.

Simultaneous multi-slice (SMS) imaging is a popular technique for increasing acquisition speed in echo-planar imaging (EPI) fMRI. However, SMS data are prone to motion sensitivity and slice leakage artefacts, which spread signal between simultaneously acquired slices. Relevant to motion sensitivity, artefacts from moving anatomic structures propagate along the phase-encoding (PE) direction. This is particularly relevant for eye movement. As signal from the eye is acquired along with signal from simultaneously excited slices during SMS, there is potential for signal to spread in-plane and between spatially remote slices. After identifying an artefact temporally coinciding with signal fluctuations in the eye and spatially distributed in correspondence with multiband slice acceleration and parallel imaging factors, we conducted a series of small experiments to investigate eye movement artefacts in SMS data and the contribution of PE direction to the invasiveness of these artefacts. Five healthy adult volunteers were scanned during a blinking task using a standard SMS-EPI protocol with posterior-to-anterior (P ≫ A), anterior-to-posterior (A ≫ P) or right-to-left (R ≫ L) PE direction. The intensity of signal fluctuations (artefact severity) was measured at expected artefact positions and control positions. We demonstrated a direct relationship between eye movements and artefact severity across expected artefact regions. Within-brain artefacts were apparent in P ≫ A- and A ≫ P-acquired data but not in R ≫ L data due to the shift in artefact positions. Further research into eye motion artefacts in SMS data is warranted but researchers should exercise caution with SMS protocols. We recommend rigorous piloting of SMS protocols and switching to R ≫ L/L ≫ R PE where feasible.

At ambient conditions, both dolomite [CaMg(CO3)2] and magnesite [MgCO3] do not readily grow from aqueous solution. A common explanation for this is the highly hydrated character of Mg2+. The exceptionally easy growth of norsethite [BaMg(CO3)2], however, clearly shows that Mg2+ ions, in principle, can rapidly dehydrate and incorporate into anhydrous carbonate minerals even at ambient conditions. Still, the lack of reliable quantitative data prevents the necessary comparison of the reactivities of these magnesium-bearing minerals. In order to shed light on the span of possible incorporation rates of anhydrous Mg2+ ions, we present the first systematic quantitative study of norsethite growth kinetics as well as the determination of norsethite solubility product over a wide range of temperatures to replace the diffuse qualitative knowledge existing so far. Norsethite solubility was determined in 0.1 M NaCl aqueous solutions from 30 to 150 °C using a hydrogen-electrode concentration cell, which provides a continuous in-situ measurement of hydrogen ion molality. The solubility product of norsethite can be described by log10Ksp°-nrs = a + b/T + cT, where a = 31.007, b = −7321.122, and c = −0.0811. Gibbs free energy ( Δ f G 298.15 0 ) and enthalpy ( Δ f H 298.15 0 ) of norsethite formation were determined to be −2167 ± 2 kJ/mol and −2351 ± 2 kJ/mol, respectively. Growth experiments were conducted in mixed-flow reactors covering a significant span of solution compositions (pH: 7.0–8.5, [Ba]: 3 × 10−6–5 × 10−3 M, [Mg]: 1 × 10−4–9 × 10−2 M, ionic strength: 0.1 M, Ωnorsethite = 1–290) and temperatures (40, 65, and 100 °C). From the experimental data, the apparent activation energy of norsethite growth rate constant was determined to be Ea = 80 ± 7 kJ/mol. An extrapolation to 25 °C resulted in a rate constant of k nrs 25 ∘ C = 1.8 × 10−2 nmol m−2 s−1 with a reaction order of 1.2 ± 0.1. These results allowed for a direct, quantitative comparison of the growth rates of different anhydrous Mg-bearing carbonate minerals. This comparison revealed that the growth rate constant of norsethite at 100 °C is approximately three orders of magnitude higher than that of magnesite and five orders of magnitude higher than that of dolomite. In the case of norsethite, obviously some effective means must exist which promotes the dehydration of the Mg ion and allows for the rapid incorporation of dehydrated Mg2+ into the growing mineral. This promotion has to take place at the norsethite surface where the hydration energy of Mg2+ can significantly differ from the well-known value in bulk solution. Consequently, not only the stability of the aqueous metal complex per se is an important factor controlling the growth rate of anhydrous magnesium bearing carbonate minerals but also the means of a given surface to weaken the stability of this complex.

The proteasome system is the main cellular protein degradation machinery and acts as a key regulator of cellular growth, survival and signaling. Tumor cells are highly dependent on its function and the proteasome has been shown to be activated in colon or breast cancer. Its regulation in lung cancer, however, is largely unknown. We here investigated the role of the proteasome activator PA200 in lung cancer to determine its so far unknown regulation in disease and to better characterize proteasome regulation in lung cancer. The specificity of PA200 antibodies was first validated by siRNA-mediated silencing of PA200 in A549 adenocarinoma cells and analysis of wildtype and PA200-/- mouse tissues by Western blotting and immunohistochemistry (IHC). IHC indicated a strong induction of PA200 protein levels in lung tumors of KrasLA2 mutant mice when compared to wildtype lungs. Analysis of human lung cancers by Western blot and IHC analysis revealed a strong upregulation of PA200 when compared to non-tumorous tissue of the same patient. A transcriptomic screen of A549 adenocarcinoma cells with PA200 silencing for 72 h revealed inactivation of key cellular processes such as cellular proliferation, migration and infiltration. We are currently screening the PA200 expression of a large set of biopsies from different lung tumor subtypes, including adenocarcinoma, squamous cell carcinoma and small cell lung cancer, in order to evaluate its regulation in different lung cancer subtypes and to correlate its expression with with the clinical data of the patients. Concluding, our findings indicate that PA200 is strongly upregulated in lung cancer tissues and suggest a function as a potential driver of tumor cell growth and invasion.

Animal studies have shown that the striatal cholinergic system plays a role in behavioural flexibility but, until recently, this system could not be studied in humans due to a lack of appropriate non-invasive techniques. Using proton magnetic resonance spectroscopy (1H-MRS) we recently showed that the concentration of dorsal striatal choline (an acetylcholine precursor) changes during reversal learning (a measure of behavioural flexibility) in humans. The aim of the present study was to examine whether regional average striatal choline was associated with reversal learning. 36 participants (mean age = 24.8, range = 18-32, 20 female) performed a probabilistic learning task with a reversal component. We measured choline at rest in both the dorsal and ventral striatum using 1H-MRS. Task performance was described using a simple reinforcement learning model that dissociates the contributions of positive and negative prediction errors to learning. Average levels of choline in the dorsal striatum were associated with performance during reversal, but not during initial learning. Specifically, lower levels of choline in the dorsal striatum were associated with a lower number of perseverative trials. Moreover, choline levels explained inter-individual variance in perseveration over and above that explained by learning from negative prediction errors. These findings suggest that the dorsal striatal cholinergic system plays an important role in behavioural flexibility, in line with evidence from the animal literature and our previous work in humans. Additionally, this work provides further support for the idea of measuring choline with 1H-MRS as a non-invasive way of studying human cholinergic neurochemistry.SIGNIFICANCE STATEMENTBehavioural flexibility is a crucial component of adaptation and survival. Evidence from the animal literature shows the striatal cholinergic system is fundamental to reversal learning, a key paradigm for studying behavioural flexibility, however, this system remains understudied in humans. Using proton magnetic resonance spectroscopy, we showed that choline levels at rest in the dorsal striatum are associated with performance specifically during reversal learning. These novel findings help to bridge the gap between animal and human studies by demonstrating the importance of cholinergic function in the dorsal striatum in human behavioural flexibility. Importantly, the methods described here can not only be applied to furthering our understanding of healthy human neurochemistry, but also to extending our understanding of cholinergic disorders.

During the last years, complex network approaches have demonstrated their great potentials as versatile tools for exploring the structural as well as dynamical properties of complex systems from a variety of different fields. Among others, recent successful examples include their application to studying flow systems in both, abstract mathematical and real-world geophysical contexts. In this context, two recent developments are particularly notable: on the one hand, correlation-based functional network approaches allow inferring statistical interrelationships, for example between macroscopic regions of the Earth’s climate system, which are hidden to more classical statistical analysis techniques. On the other hand, Lagrangian flow networks provide a new tool to identify dynamically relevant structures in atmosphere, ocean or, more generally, the phase space of complex systems. This chapter summarizes these recent developments and provides some illustrative examples highlighting the application of both concepts to selected paradigmatic low-dimensional model systems.

Animal studies have shown that acetylcholine (ACh) levels in the dorsal striatum play a role in reversal learning. However, this has not been studied in humans due to a lack of appropriate non-invasive techniques. Proton magnetic resonance spectroscopy (1H-MRS) can be used to measure metabolite levels in humans in vivo. Although it cannot be used to study ACh directly, 1H-MRS can be used to study choline, an ACh precursor which is linked to activity-dependent ACh release. The aim of this study was to use functional-1H-MRS (fMRS) to measure changes in choline levels in the human dorsal striatum during performance of a probabilistic reversal learning task. We demonstrate a task-dependent decrease in choline, specifically during reversal, but not initial, learning. We interpret this to reflect a sustained increase in ACh levels, which is in line with findings from the animal literature. This task-dependent change was specific to choline and was not observed in control metabolites. These findings provide support for the use of fMRS in the in vivo study of the human cholinergic system.

Background In idiopathic pulmonary fibrosis (IPF), fibroblasts gain a more migratory phenotype and excessively secrete extracellular matrix (ECM), ultimately leading to alveolar scarring and progressive dyspnea. Here, we analyzed the effects of deficiency of FK506-binding protein 10 (FKBP10), a potential IPF drug target, on primary human lung fibroblast (phLF) adhesion and migration. Methods Using siRNA, FKBP10 expression was inhibited in phLF in absence or presence of 2ng/ml transforming growth factor-β1 (TGF-β1) and 0.1mM 2-phosphoascorbate. Effects on cell adhesion and migration were monitored by an immunofluorescence (IF)-based attachment assay, a conventional scratch assay, and single cell tracking by time-lapse microscopy. Effects on expression of key players in adhesion dynamics and migration were analyzed by qPCR and Western Blot. Colocalization was evaluated by IF microscopy and by proximity ligation assays. Results FKBP10 knockdown significantly attenuated adhesion and migration of phLF. Expression of collagen VI was decreased, while expression of key components of the focal adhesion complex was mostly upregulated. The effects on migration were 2-phosphoascorbate-dependent, suggesting collagen synthesis as the underlying mechanism. FKBP10 colocalized with collagen VI and coating culture dishes with collagen VI, and to a lesser extent with collagen I, abolished the effect of FKBP10 deficiency on migration. Conclusions These findings show, to our knowledge for the first time, that FKBP10 interacts with collagen VI and that deficiency of FKBP10 reduces phLF migration mainly by downregulation of collagen VI synthesis. The results strengthen FKBP10 as an important intracellular regulator of ECM remodeling and support the concept of FKBP10 as drug target in IPF.

In this paper we discuss the results of an intercomparison for electric field strength measurements within the German Calibration Service (Deutscher Kalibrierdienst – DKD). The comparison has been carried out on the field strength value required to reach a display reading of 20 V m−1 of the field probes for frequencies between 100 MHz and 18 GHz. Five laboratories joined the intercomparison including the Physikalisch-Technische Bundesanstalt (PTB), the German National Metrology Institute that keeps the primary standard for electric field strength. As measurement artefacts both a small 1-axis probe usually used as transfer sensor at PTB and a larger 3-axis commercial field probe have been used. While the results agree well for the small field probe and when the larger commercial 3-axis field probe is oriented in the direction of the magnetic field, larger deviations occur, when the larger 3-axis field probe is oriented into the direction of the Poynting vector of the calibration field.

Cognitive flexibility is crucial for adaptation and is disrupted in neuropsychiatric disorders and psychopathology. Human studies of flexibility using reversal learning tasks typically contrast error trials before and after reversal, which provides little information about the mechanisms that support learning and expressing a new response. However, animal studies suggest a specific role in this latter process for the connections between the dorsal striatum and the centromedian parafascicular (CM-Pf) thalamus, a system which may recruit the striatal cholinergic interneurons, but which is not well understood in humans. This study investigated the role of this system in human probabilistic reversal learning, specifically with respect to learning a new response strategy, contrasting its function to that of the better understood orbitoftontal-striatal systems. Using psychophysiological interaction (PPI) analysis of functional magnetic resonance imaging (fMRI) data we show that connectivity between the striatum and both the lateral orbitofrontal cortex (lOFC) and CM-Pf pathways increased during reversal, but not initial learning. However, while the strength of lOFC-striatal connectivity was associated with the speed of the reversal, the strength of CM-Pf-striatal connectivity was associated specifically with the quality of the reversal (reduced regressive errors). These findings expand our understanding of flexibility mechanisms in the human brain, bridging the gap with animal studies of this system.

Cues from the extracellular matrix (ECM) and their functional interplay with cells play pivotal roles for development, tissue repair, and disease. However, the precise nature of this interplay remains elusive. We used an innovative 3D cell culture ECM model by decellularizing 300-µm-thick ex vivo lung tissue scaffolds (d3D-LTCs) derived from diseased and healthy mouse lungs, which widely mimics the native (patho)physiological in vivo ECM microenvironment. We successfully repopulated all d3D-LTCs with primary human and murine fibroblasts, and moreover, we demonstrated that the cells also populated the innermost core regions of the d3D-LTCs in a real 3D fashion. The engrafted fibroblasts revealed a striking functional plasticity, depending on their localization in distinct ECM niches of the d3D-LTCs, affecting the cells’ tissue engraftment, cellular migration rates, cell morphologies, and protein expression and phosphorylation levels. Surprisingly, we also observed fibroblasts that were homing to the lung scaffold’s interstitium as well as fibroblasts that were invading fibrotic areas. To date, the functional nature and even the existence of 3D cell matrix adhesions in vivo as well as in 3D culture models is still unclear and controversial. Here, we show that attachment of fibroblasts to the d3D-LTCs evidently occurred via focal adhesions, thus advocating for a relevant functional role in vivo. Furthermore, we found that protein levels of talin, paxillin, and zyxin and phosphorylation levels of paxillin Y118, as well as the migration-relevant small GTPases RhoA, Rac, and CDC42, were significantly reduced compared with their attachment to 2D plastic dishes. In summary, our results strikingly indicate that inherent physical or compositional characteristics of the ECM act as instructive cues altering the functional behavior of engrafted cells. Thus, d3D-LTCs might aid to obtain more realistic data in vitro, with a high relevance for drug discovery and mechanistic studies alike.

Hintergrund: Tuberkulose zahlt weltweit neben HIV und Malaria immer noch zu den haufigsten Infektionserkrankungen und stellt eine ernste Problematik im Gesundheitsbereich dar. Durch Einwanderer und Fluchtlinge ist die Erkrankung auch in Europa weiterhin prasent. Die globale Zunahme einer multiresistenten Tuberkulose fuhrt somit zu einer hoch bedeutsamen und aktuellen Wichtigkeit suffizienter Therapiemasnahmen. Dies hat in den letzten Jahren zu einer erneuten Evaluation der chirurgischen Therapie im Rahmen der interdisziplinaren und multimodalen Therapie der multiresistenten Tuberkulose gefuhrt. Zudem bestehen trotz effektiver medikamentoser Therapie der antibiotikaempfindlichen Tuberkulose weiterhin Indikationen zur chirurgischen Therapie. Neben der massiven Hamoptoe als notfallmasige Operationsindikation stehen hier vor allem tuberkulose Sekundarkomplikationen wie Aspergillome, chronische Hamoptysen, Pneumothorax, bronchopleurale Fisteln und zerstorte Areale des Lungenparenchyms („destroyed lung“) im Vordergrund. Schlussfolgerung: Die interdisziplinare OP-Indikation sollte stets in einem multimodalen Therapiekonzept unter Berucksichtigung von Patientenalter und Funktionsanalysen gestellt werden. Eine antituberkulostatische Therapie sollte vor und nach einer Operation suffizient durchgefuhrt werden, um einen nachhaltigen Therapieerfolg verzeichnen zu konnen.

Chronic obstructive pulmonary disease (COPD) is characterised by a progressive loss of lung tissue. Inducing repair processes within the adult diseased lung is of major interest and Wnt/β-catenin signalling represents a promising target for lung repair. However, the translation of novel therapeutic targets from model systems into clinical use remains a major challenge.We generated murine and patient-derived three-dimensional (3D) ex vivo lung tissue cultures (LTCs), which closely mimic the 3D lung microenvironment in vivo. Using two well-known glycogen synthase kinase-3β inhibitors, lithium chloride (LiCl) and CHIR 99021 (CT), we determined Wnt/β-catenin-driven lung repair processes in high spatiotemporal resolution using quantitative PCR, Western blotting, ELISA, (immuno)histological assessment, and four-dimensional confocal live tissue imaging.Viable 3D-LTCs exhibited preserved lung structure and function for up to 5 days. We demonstrate successful Wnt/β-catenin signal activation in murine and patient-derived 3D-LTCs from COPD patients. Wnt/β-catenin signalling led to increased alveolar epithelial cell marker expression, decreased matrix metalloproteinase-12 expression, as well as altered macrophage activity and elastin remodelling. Importantly, induction of surfactant protein C significantly correlated with disease stage (per cent predicted forced expiratory volume in 1 s) in patient-derived 3D-LTCs.Patient-derived 3D-LTCs represent a valuable tool to analyse potential targets and drugs for lung repair. Enhanced Wnt/β-catenin signalling attenuated pathological features of patient-derived COPD 3D-LTCs.

Nanoparticles allow for controlled and targeted drug delivery to diseased tissues and therefore bypass systemic side effects. Spatiotemporal control of drug release can be achieved by nanocarriers that respond to elevated levels of disease-specific enzymes. For example, matrix metalloproteinase 9 (MMP9) is overexpressed in tumors, is known to enhance the metastatic potency of malignant cells, and has been associated with poor prognosis of lung cancer. Here, we report the synthesis of mesoporous silica nanoparticles (MSNs) tightly capped by avidin molecules via MMP9 sequence-specific linkers to allow for site-selective drug delivery in high-expressing MMP9 tumor areas. We provide proof-of-concept evidence for successful MMP9-triggered drug release from MSNs in human tumor cells and in mouse and human lung tumors using the novel technology of ex vivo 3D lung tissue cultures. This technique allows for translational testing of drug delivery strategies in diseased mouse and human tissue. Using this method we show MMP9-mediated release of cisplatin, which induced apoptotic cell death only in lung tumor regions of Kras mutant mice, without causing toxicity in tumor-free areas or in healthy mice. The MMP9-responsive nanoparticles also allowed for effective combinatorial drug delivery of cisplatin and proteasome inhibitor bortezomib, which had a synergistic effect on the (therapeutic) efficiency. Importantly, we demonstrate the feasibility of MMP9-controlled drug release in human lung tumors.

Leiomyoma of the pulmonary artery represents a curiosity in the literature. We describe a case of a 54-year-old female patient who presented with recurrent cough of a few weeks' duration. Computed tomography of the thorax located a smooth, limited tumor in the left thorax near the interlobar space. Thoracoscopic exploration showed a tumor mass, fused with the pulmonary artery. After anterolateral thoracotomy, a complete resection of the tumor was performed. The histopathologic examination showed the presence of a vascular leiomyoma of the tunica media of the pulmonary artery.

Background: DLPD are frequently detected and better evaluated by means of current diagnostic guidelines. Methods: Prospective assessment of patients with DLPD referred to a regional reference center in Germany using clinical, functional and radiological parameters, and results of bronchoscopic or surgical results. The diagnosis was made after a multidisciplinary discussion (MDD). Results: Between January and June of 2016 342 patients were admitted to the hospital for diagnosis and therapy of DPLD (144 women, average age 63,5 years), on average with a mild restrictive ventilatory defects (O VC 76,1 %, O DLCOcSB 61,1 %). IPF was the subgroup detected with the lowest DLCOcSB (O 46,7 %), pneumoconiosis showed the highest results (O 87,2 %); alveolar proteinosis showed the lowest VC (O 63,0 %), LAM showed the highest VC (O 93,3 %). 50 patients were diagnosed with the help of clinical and radiological findings. 292 patients underwent a bronchoscopy, in 133 cases a transbronchial cryobiopsy was performed. A surgical lung biopsy for the diagnosis of DPLD was performed in 20 patients. In 323 cases a precise diagnose could be obtained after MDD (94,4%): IPF (n= 59), sarcoidosis (n=50), hypersensitivity pneumonitis (n=47), collagen vascular disease (n=36), organizing pneumonia (n=32), nonspecific interstitial pneumonia (n=23), pneumoconiosis (n=14), other DLPD DLPD (n=62). In 19 cases a specific diagnose could not be obtained. Conclusion: Using a standardised approach including bronchoscopic assessment and the MDD, a specific diagnosis of DPLD can be made in 94 % of cases. A robust histological result could be obtained in 63% of patients using transbronchial cryobiopsies. The most prevalent DLPD are IPF, sarcoidosis and hypersensitivity pneumonitis.