Charakterisierung des Migrationsverhaltens von pankreatischen duktalen Adenokarzinomzellen bei der Comigration mit pankreatischen Sternzellen

Pankreatische Sternzellen (PSC) spielen eine tragende Rolle bei der Progression und Invasivität des duktalen Adenokarzinoms des Pankreas (PDAC). Ziel dieser Arbeit war die Charakterisierung des Migrationsverhaltens von PDAC-Zellen bei der Comigration mit PSC und der Beteiligung von ausgewählten Proteinen. Dazu wurden Videomikroskopie von einzelnen Zellen und Zellverbänden, proteinanalytische Methoden und Messungen der intrazellulären Calciumkonzentration durchgeführt. Eine Steigerung der Zellmigration von vereinzelten PDAC-Zellen zeigte sich unter Einfluss von EGF, konditioniertem Medium von PSC-Kulturen und durch Inhibition des KCa3.1-Kaliumkanals. Eine Verringerung der PDAC-Motilität ergab sich durch Inhibition des Na+/H+-Austauschers NHE1. Die An- oder Abwesenheit von PSC führte bei PDAC-Zellverbänden zu keiner Änderung der Migrationsaktivität und PDAC-Zellen waren umgekehrt nicht zur Stimulation eines PSC-Verbandes in der Lage.<br

Data Governance and Sovereignty in Urban Data Spaces Based on Standardized ICT Reference Architectures

European cities and communities (and beyond) require a structured overview and a set of tools as to achieve a sustainable transformation towards smarter cities/municipalities, thereby leveraging on the enormous potential of the emerging data driven economy. This paper presents the results of a recent study that was conducted with a number of German municipalities/cities. Based on the obtained and briefly presented recommendations emerging from the study, the authors propose the concept of an Urban Data Space (UDS), which facilitates an eco-system for data exchange and added value creation thereby utilizing the various types of data within a smart city/municipality. Looking at an Urban Data Space from within a German context and considering the current situation and developments in German municipalities, this paper proposes a reasonable classification of urban data that allows the relation of various data types to legal aspects, and to conduct solid considerations regarding technical implementation designs and decisions. Furthermore, the Urban Data Space is described/analyzed in detail, and relevant stakeholders are identified, as well as corresponding technical artifacts are introduced. The authors propose to setup Urban Data Spaces based on emerging standards from the area of ICT reference architectures for Smart Cities, such as DIN SPEC 91357 “Open Urban Platform” and EIP SCC. In the course of this, the paper walks the reader through the construction of a UDS based on the above-mentioned architectures and outlines all the goals, recommendations and potentials, which an Urban Data Space can reveal to a municipality/city. Finally, we aim at deriving the proposed concepts in a way that they have the potential to be part of the required set of tools towards the sustainable transformation of German and European cities in the direction of smarter urban environments, based on utilizing the hidden potential of digitalization and efficient interoperable data exchange.EC/H2020/646578/EU/Triangulum: The Three Point Project / Demonstrate. Disseminate. Replicate./TriangulumBMBF, 13NKE012, Datenaustausch und Zusammenarbeit im urbanen Raum - Bestandsanalyse (Urban Data Space

Nutritional status of flexitarians compared to vegans and omnivores - a cross-sectional pilot study

Background: In the Western world, there has been a notable rise in the popularity of plant-based, meat-reduced flexitarian diets. Nevertheless, there is insufficient data on the nutritional status of individuals following this dietary pattern. The aim of this study was to investigate the intake and endogenous status of various nutrients in a healthy German adult study population consisting of flexitarians (FXs), vegans (Vs) and omnivores (OMNs). Methods: In this cross-sectional study, dietary intake of 94 non-smoking adults (32 FXs, 33 Vs, 29 OMNs) between 25 and 45 years of age was assessed using 3-day dietary records. In addition, blood samples were collected to determine different endogenous nutrient status markers. Results: 32%, 82% and 24% of the FXs, Vs, and OMNs respectively reported using dietary supplements. In the FXs, intake of total energy as well as macronutrients and most micronutrients were within the reference range. FXs had higher intakes of fiber, retinol-equ., ascorbic acid, folate-equ., tocopherol-equ., calcium, and magnesium compared to OMNs. However, cobalamin intake in FXs (2.12 µg/d) was below the reference (4 µg/d). Based on 4cB12, 13% of FXs showed a cobalamin undersupply [< -0.5 to -2.5] compared to 10% of OMNs, and 9% of Vs. The median 25(OH)D serum concentrations in FXs, Vs and OMNs were 46.6, 55.6, and 59.6 nmol/L. The prevalence of an insufficient/deficient vitamin-D status [< 49.9 nmol 25(OH)D/L] was highest in FXs (53%), followed by Vs (34%) and OMNs (27%). In FXs and Vs, the supplement takers had better cobalamin and vitamin-D status than non-supplement takers. Anemia and depleted iron stores were found only occasionally in all groups. In women, the prevalence of pre-latent iron deficiency and iron deficiency was highest in FXs (67%) compared to Vs (61%) and OMNs (54%). Conclusion: Our findings indicated that all three diets delivered sufficient amounts of most macro- and micronutrients. However, deficiencies in cobalamin, vitamin-D, and iron status were common across all diets. Further studies are needed to investigate the nutrient supply status and health consequences of meat-reduced plant-based diets. The study was registered in the German Clinical Trial Register (number: DRKS 00019887, data: 08.01.2020)

A Novel NHE1-Centered Signaling Cassette Drives Epidermal Growth Factor Receptor–Dependent Pancreatic Tumor Metastasis and Is a Target for Combination Therapy

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers principally because of early invasion and metastasis. The epidermal growth factor receptor (EGFR) is essential for PDAC development even in the presence of Kras, but its inhibition with erlotinib gives only a modest clinical response, making the discovery of novel EGFR targets of critical interest. Here, we revealed by mining a human pancreatic gene expression database that the metastasis promoter Na+/H+ exchanger (NHE1) associates with the EGFR in PDAC. In human PDAC cell lines, we confirmed that NHE1 drives both basal and EGF-stimulated three-dimensional growth and early invasion via invadopodial extracellular matrix digestion. EGF promoted the complexing of EGFR with NHE1 via the scaffolding protein Na&nbsp;+/H&nbsp;+ exchanger regulatory factor 1, engaging EGFR in a negative transregulatory loop that controls the extent and duration of EGFR oncogenic signaling and stimulates NHE1. The specificity of NHE1 for growth or invasion depends on the segregation of the transient EGFR/Na&nbsp;+/H&nbsp;+ exchanger regulatory factor 1/NHE1 signaling complex into dimeric subcomplexes in different lipid raftlike membrane domains. This signaling complex was also found in tumors developed in orthotopic mice. Importantly, the specific NHE1 inhibitor cariporide reduced both three-dimensional growth and invasion independently of PDAC subtype and synergistically sensitized these behaviors to low doses of erlotinib

A Potent and Selective S1P1 Antagonist with Efficacy in Experimental Autoimmune Encephalomyelitis

SummaryLymphocyte trafficking is critically regulated by the Sphingosine 1-phosphate receptor-1 (S1P1), a G protein-coupled receptor that has been highlighted as a promising therapeutic target in autoimmunity. Fingolimod (FTY720, Gilenya) is a S1P1 receptor agonist that has recently been approved for the treatment of multiple sclerosis (MS). Here, we report the discovery of NIBR-0213, a potent and selective S1P1 antagonist that induces long-lasting reduction of peripheral blood lymphocyte counts after oral dosing. NIBR-0213 showed comparable therapeutic efficacy to fingolimod in experimental autoimmune encephalomyelitis (EAE), a model of human MS. These data provide convincing evidence that S1P1 antagonists are effective in EAE. In addition, the profile of NIBR-0213 makes it an attractive candidate to further study the consequences of S1P1 receptor antagonism and to differentiate the effects from those of S1P1 agonists

Genetic diversity of the arbuscular mycorrhizal fungus Glomus intraradices as determined by mitochondrial large subunit rRNA gene sequences is considerably higher than previously expected

Summary: • Glomus intraradices is a widespread arbuscular mycorrhizal fungus (AMF), which has been found in an extremely broad range of habitats, indicating a high tolerance for environmental factors and a generalist life history strategy. Despite this ecological versatility, not much is known about the genetic diversity of this fungal species across different habitats or over large geographic scales. • A nested polymerase chain reaction (PCR) approach for the mitochondrial rRNA large subunit gene (mtLSU), distinguished different haplotypes among cultivated isolates of G. intraradices and within mycorrhizal root samples from the field. • From analysis of 16 isolates of this species originating from five continents, 12 mitochondrial haplotypes were distinguished. Five additional mtLSU haplotypes were detected in field‐collected mycorrhizal roots. Some introns in the mtLSU region appear to be stable over years of cultivation and are ancestral to the G. intraradices clade. • Genetic diversity within G. intraradices is substantially higher than previously thought, although some mtLSU haplotypes are widespread. A restriction fragment length polymorphism approach also was developed to distinguish mtLSU haplotypes without sequencing. Using this molecular tool, intraspecific genetic variation of an AMF species can be studied directly in field plants

Ring1b-dependent epigenetic remodelling is an essential prerequisite for pancreatic carcinogenesis

BACKGROUND AND AIMS Besides well-defined genetic alterations, the dedifferentiation of mature acinar cells is an important prerequisite for pancreatic carcinogenesis. Acinar-specific genes controlling cell homeostasis are extensively downregulated during cancer development; however, the underlying mechanisms are poorly understood. Now, we devised a novel in vitro strategy to determine genome-wide dynamics in the epigenetic landscape in pancreatic carcinogenesis. DESIGN With our in vitro carcinogenic sequence, we performed global gene expression analysis and ChIP sequencing for the histone modifications H3K4me3, H3K27me3 and H2AK119ub. Followed by a comprehensive bioinformatic approach, we captured gene clusters with extensive epigenetic and transcriptional remodelling. Relevance of Ring1b-catalysed H2AK119ub in acinar cell reprogramming was studied in an inducible Ring1b knockout mouse model. CRISPR/Cas9-mediated Ring1b ablation as well as drug-induced Ring1b inhibition were functionally characterised in pancreatic cancer cells. RESULTS The epigenome is vigorously modified during pancreatic carcinogenesis, defining cellular identity. Particularly, regulatory acinar cell transcription factors are epigenetically silenced by the Ring1b-catalysed histone modification H2AK119ub in acinar-to-ductal metaplasia and pancreatic cancer cells. Ring1b knockout mice showed greatly impaired acinar cell dedifferentiation and pancreatic tumour formation due to a retained expression of acinar differentiation genes. Depletion or drug-induced inhibition of Ring1b promoted tumour cell reprogramming towards a less aggressive phenotype. CONCLUSIONS Our data provide substantial evidence that the epigenetic silencing of acinar cell fate genes is a mandatory event in the development and progression of pancreatic cancer. Targeting the epigenetic repressor Ring1b could offer new therapeutic options

Subunit composition, molecular environment, and activation of native TRPC channels encoded by their interactomes.

peer reviewedIn the mammalian brain TRPC channels, a family of Ca2+-permeable cation channels, are involved in a variety of processes from neuronal growth and synapse formation to transmitter release, synaptic transmission and plasticity. The molecular appearance and operation of native TRPC channels, however, remained poorly understood. Here, we used high-resolution proteomics to show that TRPC channels in the rodent brain are macro-molecular complexes of more than 1 MDa in size that result from the co-assembly of the tetrameric channel core with an ensemble of interacting proteins (interactome). The core(s) of TRPC1-, C4-, and C5-containing channels are mostly heteromers with defined stoichiometries for each subtype, whereas TRPC3, C6, and C7 preferentially form homomers. In addition, TRPC1/C4/C5 channels may co-assemble with the metabotropic glutamate receptor mGluR1, thus guaranteeing both specificity and reliability of channel activation via the phospholipase-Ca2+ pathway. Our results unveil the subunit composition of native TRPC channels and resolve the molecular details underlying their activation

Baseline Chest Computed Tomography as Standard of Care in High-Risk Hematology Patients

Baseline chest computed tomography (BCT) in high-risk hematology patients allows for the early diagnosis of invasive pulmonary aspergillosis (IPA). The distribution of BCT implementation in hematology departments and impact on outcome is unknown. A web-based questionnaire was designed. International scientific bodies were invited. The estimated numbers of annually treated hematology patients, chest imaging timepoints and techniques, IPA rates, and follow-up imaging were assessed. In total, 142 physicians from 43 countries participated. The specialties included infectious diseases (n = 69; 49%), hematology (n = 68; 48%), and others (n = 41; 29%). BCT was performed in 57% (n = 54) of 92 hospitals. Upon the diagnosis of malignancy or admission, 48% and 24% performed BCT, respectively, and X-ray was performed in 48% and 69%, respectively. BCT was more often used in hematopoietic cell transplantation and in relapsed acute leukemia. European centers performed BCT in 59% and non-European centers in 53%. Median estimated IPA rate was 8% and did not differ between BCT (9%; IQR 5-15%) and non-BCT centers (7%; IQR 5-10%) (p = 0.69). Follow-up computed tomography (CT) for IPA was performed in 98% (n = 90) of centers. In high-risk hematology patients, baseline CT is becoming a standard-of-care. Chest X-ray, while inferior, is still widely used. Randomized, controlled trials are needed to investigate the impact of BCT on patient outcome