Phenotypic characterization of aberrant stem and progenitor cell populations in myelodysplastic syndromes

Recent reports have revealed myelodysplastic syndromes (MDS) to arise from cancer stem cells phenotypically similar to physiological hematopoietic stem cells. Myelodysplastic hematopoiesis maintains a hierarchical organization, but the proportion of several hematopoietic compartments is skewed and multiple surface markers are aberrantly expressed. These aberrant antigen expression patterns hold diagnostic and therapeutic promise. However, eradication of MDS requires targeting of early myelodysplasia propagating stem cells. This warrants an exact assessment of the differentiation stage at which aberrant expression occurs in transformed hematopoiesis. Here, we report results on the prospective and extensive dissection of the hematopoietic hierarchy in 20 patients with either low-risk MDS or MDS with excess blasts and compare it to hematopoiesis in patients with non-malignancy associated cytopenia or B cell lymphoma without bone marrow infiltration. We found patients with MDS with excess blasts to exhibit characteristic expansions of specific immature progenitor compartments. We also identified the aberrant expression of several markers including ALDH, CLL-1, CD44, and CD47 to be specific features of hematopoiesis in MDS with excess blasts. We show that amongst these, aberrant CLL-1 expression manifested at the early uncommitted hematopoietic stem cell level, suggesting a potential role as a therapeutic target

CRP-cAMP mediates silencing of Salmonella virulence at the post-transcriptional level

Invasion of epithelial cells by Salmonella enterica requires expression of genes located in the pathogenicity island I (SPI-1). The expression of SPI-1 genes is very tightly regulated and activated only under specific conditions. Most studies have focused on the regulatory pathways that induce SPI-1 expression. Here, we describe a new regulatory circuit involving CRP-cAMP, a widely established metabolic regulator, in silencing of SPI-1 genes under non-permissive conditions. In CRP-cAMP-deficient strains we detected a strong upregulation of SPI-1 genes in the mid-logarithmic growth phase. Genetic analyses revealed that CRP-cAMP modulates the level of HilD, the master regulator of Salmonella invasion. This regulation occurs at the post-transcriptional level and requires the presence of a newly identified regulatory motif within the hilD 3’UTR. We further demonstrate that in Salmonella the Hfq-dependent sRNA Spot 42 is under the transcriptional repression of CRP-cAMP and, when this transcriptional repression is relieved, Spot 42 exerts a positive effect on hilD expression. In vivo and in vitro assays indicate that Spot 42 targets, through its unstructured region III, the 3’UTR of the hilD transcript. Together, our results highlight the biological relevance of the hilD 3’UTR as a hub for post-transcriptional control of Salmonella invasion gene expression.Spanish Ministry of Economy and Competitiveness BIO2010-15417 BIO2013-44220-R AGL2013-45339-RRecerCaixa program 2012/ACUP/00048Catalonian government 2017SGR49

Bildung einer Fiskalunion: Ein wirkungsvolles Instrument zur Stabilisierung der Eurozone?

Im Juni 2015 legten Jean-Claude Juncker, Donald Tusk, Jeroen Dijsselbloem, Mario Draghi und Martin Schulz Vorschläge zur »langfristigen Stärkung der Währungsunion« vor. Sie sehen unter anderem auch die Einsetzung eines beratenden Europäischen Fiskalausschusses vor, der die Arbeit der nationalen Räte für Finanzpolitik koordinieren soll. Kann das ein wirkungsvolles Instrument zur Stabilisierung der Eurozone sein? Nach Ansicht von Charles B. Blankart, Humboldt-Universität zu Berlin und Universität Luzern, hat sich die Europäische Union seit etwa 1992 grundlegend verändert. Während sie bis dahin dem zweiseitigen Handel, einem privaten Gut diente, stehe seither der Euro als ein öffentliches Gut im Vordergrund. Blankart sieht in den Zielen der Vorschläge die bekannten »Öffentlichen-Gut-Probleme« des Euro. Die Vorleistungen des einen bewirken nicht automatisch Gegenleistungen des anderen. Das Freifahrerverhalten wird nicht beseitigt. Katharina Gnath, Bertelsmann Stiftung, und Jörg Haas, Jacques Delors Institut – Berlin, argumentieren, dass eine Fiskalunion nur dann stabilisieren kann, wenn sie sowohl glaubwürdige Haushaltsregeln als auch Elemente der Teilung von Risiko umfasst. Sie sei ein wichtiges Mittel zur wirksamen Stärkung der Eurozone, dürfe aber nicht das einzige bleiben. Thiess Büttner, Universität Erlangen-Nürnberg, ist der Ansicht, dass der Vorschlag kaum geeignet sei, die Europäische Währungsunion durchgreifend zu stabilisieren. Je nach der Ausgestaltung könnte er sich sogar als kontraproduktiv erweisen. Frank Westermann, Universität Osnabrück, sieht Parallelen in der jüngsten Entwicklung der Europäischen Währungsunion zu derjenigen Ostdeutschlands nach der Wiedervereinigung. Die reale Lohnentwicklung eile der Produktivität voraus, und es entwickle sich ein Transfersystem, das einen Aufschub notwendiger Arbeitsmarktreformen ermögliche. Eine langfristig expansive Fiskalpolitik, die in einer Fiskalunion ins­titutionalisiert würde, könne nur kurzfristig die Nachf

Dual RNA-seq of pathogen and host

Abstract | A comprehensive understanding of host-pathogen interactions requires a knowledge of the associated gene expression changes in both the pathogen and the host. Traditional, probe-dependent approaches using microarrays or reverse transcription PCR typically require the pathogen and host cells to be physically separated before gene expression analysis. However, the development of the probe-independent RNA sequencing (RNA-seq) approach has begun to revolutionize transcriptomics. Here, we assess the feasibility of taking transcriptomics one step further by performing 'dual RNA-seq', in which gene expression changes in both the pathogen and the host are analysed simultaneously. R E V I E W S 618 | SEPTEMBER 2012 | VOLUME 10 www.nature.com/reviews/micro R E V I E W S © 2012 Macmillan Publishers Limited. All rights reserved Nature Reviews | Microbiology Compare with target genome (or genomes), and enumerate sequence

A comparison of constitutive models for describing the flow of uncured styrene-butadiene rubber

Uncured styrene-butadiene rubber (SBR) can be modelled as a viscoelastic material with at least two different relaxation mechanisms. In this paper we compare multi-mode constitutive models combining two viscoelastic modes (linear and/or nonlinear) in three possible ways. Our particular choice of the two modes was inspired by models originally developed to describe the response of asphalt binders. We select the model that best fits the experimental data obtained from a modified stress relaxation experiment in the torsional configuration of the plate-plate rheometer. The optimisation of the five model parameters for each model is achieved by minimising the weighted least-squares distance between experimental observations and the computer model output using a tree-structured Parzen estimator algorithm to find an initial guess, followed by further optimisation using the Nelder-Mead simplex algorithm. The results show that the model combining the linear mode and the nonlinear mode is the most suitable variant to describe the observed behavior of SBR in the given regime. The predictive capabilities of the three models are further examined in changed experimental and numerical configurations. Full data and code to produce the figures in this article are included as supplementary material

miR-9-5p in Nephrectomy Specimens is a Potential Predictor of Primary Resistance to First-Line Treatment with Tyrosine Kinase Inhibitors in Patients with Metastatic Renal Cell Carcinoma

Approximately 20-30% of patients with metastatic renal cell carcinoma (mRCC) in first-line treatment with tyrosine kinase inhibitors (TKIs) do not respond due to primary resistance to this drug. At present, suitable robust biomarkers for prediction of a response are not available. Therefore, the aim of this study was to evaluate a panel of microRNAs (miRNAs) in nephrectomy specimens for use as predictive biomarkers for TKI resistance. Archived formalin-fixed, paraffin embedded nephrectomy samples from 60 mRCC patients treated with first-line TKIs (sunitinib, n = 51; pazopanib, n = 6; sorafenib, n = 3) were categorized into responders and non-responders. Using the standard Response Evaluation Criteria in Solid Tumors, patients with progressive disease within 3 months after the start of treatment with TKI were considered as non-responders and those patients with stable disease and complete or partial response under the TKI treatment for at least 6 months as responders. Based on a miRNA microarray expression profile in the two stratified groups of patients, seven differentially expressed miRNAs were validated using droplet digital reverse-transcription quantitative real-time polymerase chain reaction (RT-qPCR) assays in the two groups. Receiver operating characteristic curve analysis and binary logistic regression of response prediction were performed. MiR-9-5p and miR-489-3p were able to discriminate between the two groups. MiR-9-5p, as the most significant miRNA, improved the correct prediction of primary resistance against TKIs in comparison to that of conventional clinicopathological variables. The results of the decision curve analyses, Kaplan-Meier analyses and Cox regression analyses confirmed the potential of miR-9-5p in the prediction of response to TKIs and the prediction of progression-free survival after the initiation of TKI treatment

Experimental separation of the onset of slip and sharkskin melt instabilities during the extrusion of silica‑filled, styrene–butadiene rubber compounds

The flow curves of polymers often reveal the onset of melt instabilities such as sharkskin, stick–slip, or gross melt fracture, in order of increasing shear rates. The focus of this work lies in the application of the Göttfert sharkskin option to the investigation of flow curves of styrene-butadiene rubber (SBR) compounds. The sharkskin option consists of highly sensitive pressure transducers located inside a slit die of a capillary rheometer. This tool allows the detection of in-situ pressure fluctuation characteristics of different melt instabilities. It is shown that the change of slope of the transition region in the flow curves is only linked to slip. Dynamic Mechanical Analysis (DMA) measurements furthermore show that the shear rate at which the change of slope can be observed shows the same temperature dependency as the viscous and elastic properties of the compounds

Successful long-term monotherapy with rituximab in a patient with chronic lymphocytic leukemia of the B-cell-lineage: a case report

<p>Abstract</p> <p>Introduction</p> <p>Treatment of chronic lymphocytic leukemia of the B-cell-lineage is strongly based upon clinical staging because of the heterogeneous clinical course of this disease.</p> <p>Case presentation</p> <p>We describe a 62-year-old patient with newly diagnosed chronic lymphocytic leukemia of the B-cell-lineage who did not respond to several chemotherapy regimens including chlorambucil, fludarabine and cyclophosphamide, developing a marked neutropenia and thrombocytopenia with life-threatening infections. Further chemotherapy appeared not feasible because of bone marrow toxicity. The patient was treated with 600 mg/m²rituximab weekly followed by eight courses of biweekly therapy and then by long-term maintenance therapy, achieving almost complete remission of the symptoms and disease control.</p> <p>Conclusion</p> <p>After resistance to standard chemotherapy with chlorambucil and fludarabine, a patient with chronic lymphocytic leukemia of the B-cell-lineage was successfully treated with rituximab.</p

Proteinase-activated receptor 2 (PAR2) in hepatic stellate cells – evidence for a role in hepatocellular carcinoma growth in vivo

Background Previous studies have established that proteinase-activated receptor 2 (PAR2) promotes migration and invasion of hepatocellular carcinoma (HCC) cells, suggesting a role in HCC progression. Here, we assessed the impact of PAR2 in HCC stromal cells on HCC growth using LX-2 hepatic stellate cells (HSCs) and Hep3B cells as model. Methods PAR2 expression and function in LX-2 cells was analysed by RT-PCR, confocal immunofluorescence, electron microscopy, and [Ca2+]i measurements, respectively. The impact of LX-2-expressed PAR2 on tumour growth in vivo was monitored using HCC xenotransplantation experiments in SCID mice, in which HCC-like tumours were induced by coinjection of LX-2 cells and Hep3B cells. To characterise the effects of PAR2 activation in LX-2 cells, various signalling pathways were analysed by immunoblotting and proteome profiler arrays. Results Following verification of functional PAR2 expression in LX-2 cells, in vivo studies showed that these cells promoted tumour growth and angiogenesis of HCC xenografts in mice. These effects were significantly reduced when F2RL1 (encoding PAR2) was downregulated by RNA interference (RNAi). In vitro studies confirmed these results demonstrating RNAi mediated inhibition of PAR2 attenuated Smad2/3 activation in response to TGF-β1 stimulation in LX-2 cells and blocked the pro-mitotic effect of LX-2 derived conditioned medium on Hep3B cells. Furthermore, PAR2 stimulation with trypsin or a PAR2-selective activating peptide (PAR2-AP) led to activation of different intracellular signalling pathways, an increased secretion of pro-angiogenic and pro-mitotic factors and proteinases, and an enhanced migration rate across a collagen- coated membrane barrier. Silencing F2RL1 by RNAi or pharmacological inhibition of Src, hepatocyte growth factor receptor (Met), platelet-derived growth factor receptor (PDGFR), p42/p44 mitogen activated protein kinase (MAPK) or matrix-metalloproteinases (MMPs) blocked PAR2-AP-induced migration. Conclusion PAR2 in HSCs plays a crucial role in promoting HCC growth presumably by mediating migration and secretion of pro-angiogenic and pro-mitotic factors. Therefore, PAR2 in stromal HSCs may have relevance as a therapeutic target of HCC