Theratyping of the Rare CFTR Variants E193K and R334W in Rectal Organoid-Derived Epithelial Monolayers

Background: The effect of presently available CFTR modulator combinations, such as elexacaftor (ELX), tezacaftor (TEZ), and ivacaftor (IVA), on rare CFTR alleles is often unknown. Several assays have been developed, such as forskolin-induced swelling (FIS), to evaluate the rescue of such uncommon CFTR alleles both by established and novel modulators in patient-derived primary cell cultures (organoids). Presently, we assessed the CFTR-mediated electrical current across rectal organoid-derived epithelial monolayers. This technique, which allows separate measurement of CFTR-dependent chloride or bicarbonate transport, was used to assess the effect of ELX/TEZ/IVA on two rare CFTR variants. Methods: Intestinal organoid cultures were established from rectal biopsies of CF patients carrying the rare missense mutations E193K or R334W paired with F508del. The effect of the CFTR modulator combination ELX/TEZ/IVA on CFTR-mediated Cl- and HCO3- secretion was assessed in organoid-derived intestinal epithelial monolayers. Non-CF organoids were used for comparison. Clinical biomarkers (sweat chloride, FEV1) were monitored in patients receiving modulator therapy. Results: ELX/TEZ/IVA markedly enhanced CFTR-mediated bicarbonate and chloride transport across intestinal epithelium of both patients. Consistent with the rescue of CFTR function in cultured intestinal cells, ELX/TEZ/IVA therapy improved biomarkers of CFTR function in the R334W/F508del patient. Conclusions: Current measurements in organoid-derived intestinal monolayers can readily be used to monitor CFTR-dependent epithelial Cl- and HCO3- transport. This technique can be explored to assess the functional consequences of rare CFTR mutations and the efficacy of CFTR modulators. We propose that this functional CFTR assay may guide personalized medicine in patients with CF-like clinical manifestations as well as in those carrying rare CFTR mutations

An imaging system for microbial corrosion analysis

This paper describes a viable and self-contained imaging system able to assess and quantify the effects of microbial corrosion on metals surface. The proposed image processing uses Scanning Electron Microscope micrographs to analyze bacteria attachment on sample surface and to estimate the degree of degradation of the material. After a preliminary brightness and contrast normalization, which refines the image taken by the operator, the software is able to identify dark spots on the clear metal surface. These are then attributed to singly attached bacteria or to larger clusters, which are the most dangerous ones, as they could overlay corrosion pits. After that, the degradation of the material is evaluated through the quantification of microbial attachment on the surface and through dimensional distribution of bacteria clusters

The Interaction between Reactive Peritoneal Mesothelial Cells and Tumor Cells via Extracellular Vesicles Facilitates Colorectal Cancer Dissemination

Simple SummaryEmerging evidence has suggested that cancer-derived extracellular vesicles (EVs) have a crucial role in mediating directional metastasis to the peritoneal surface in colorectal cancer (CRC). We investigated the EV-mediated crosstalk between tumor and mesothelial cells which may drive remodeling of the premetastatic niche to allow tumor spread to the peritoneal surface. Our findings demonstrated that cancer-derived EVs triggered apoptosis and reduced mesothelial cell invasiveness and mesothelial-to-mesenchymal transition. On the other hand, mesothelial cells actively supported tumor invasion by releasing EVs, which induced upregulation of the major pro-invasive system in tumor cells. For the first time, we provide evidence of EV-driven mechanisms of CRC progression in patient-derived models, highlighting the crucial role of EVs in the reprogramming of mesothelial and tumor cells to establish the metastatic process.Advanced colorectal cancer (CRC) is highly metastatic and often results in peritoneal dissemination. The extracellular vesicles (EVs) released by cancer cells in the microenvironment are important mediators of tumor metastasis. We investigated the contribution of EV-mediated interaction between peritoneal mesothelial cells (MCs) and CRC cells in generating a pro-metastatic environment in the peritoneal cavity. Peritoneal MCs isolated from peritoneal lavage fluids displayed high CD44 expression, substantial mesothelial-to-mesenchymal transition (MMT) and released EVs that both directed tumor invasion and caused reprogramming of secretory profiles by increasing TGF-beta 1 and uPA/uPAR expression and MMP-2/9 activation in tumor cells. Notably, the EVs released by tumor cells induced apoptosis by activating caspase-3, peritoneal MC senescence, and MMT, thereby augmenting the tumor-promoting potential of these cells in the peritoneal cavity. By using pantoprazole, we reduced the biogenesis of EVs and their pro-tumor functions. In conclusion, our findings provided evidence of underlying mechanisms of CRC dissemination driven by the interaction of peritoneal MCs and tumor cells via the EVs released in the peritoneal cavity, which may have important implications for the clinical management of patients

Elliptic and hyperelliptic magnetohydrodynamic equilibria

The present study is a continuation of a previous one on "hyperelliptic" axisymmetric equilibria started in [Tasso and Throumoulopoulos, Phys. Plasmas 5, 2378 (1998)]. Specifically, some equilibria with incompressible flow nonaligned with the magnetic field and restricted by appropriate side conditions like "isothermal" magnetic surfaces, "isodynamicity" or P + B^2/2 constant on magnetic surfaces are found to be reducible to elliptic integrals. The third class recovers recent equilibria found in [Schief, Phys. Plasmas 10, 2677 (2003)]. In contrast to field aligned flows, all solutions found here have nonzero toroidal magnetic field on and elliptic surfaces near the magnetic axis.Comment: 9 page

Exploring the galaxy cluster-group transition regime at high redshifts: Physical properties of two newly detected z > 1 systems

Context: Multi-wavelength surveys for clusters of galaxies are opening a window on the elusive high-redshift (z>1) cluster population. Well controlled statistical samples of distant clusters will enable us to answer questions about their cosmological context, early assembly phases and the thermodynamical evolution of the intracluster medium. Aims: We report on the detection of two z>1 systems, XMMU J0302.2-0001 and XMMU J1532.2-0836, as part of the XMM-Newton Distant Cluster Project (XDCP) sample. We investigate the nature of the sources, measure their spectroscopic redshift and determine their basic physical parameters. Methods: The results of the present paper are based on the analysis of XMM-Newton archival data, optical/near-infrared imaging and deep optical follow-up spectroscopy of the clusters. Results: We confirm the X-ray source XMMU J0302.2-0001 as a gravitationally bound, bona fide cluster of galaxies at spectroscopic redshift z=1.185. We estimate its M500 mass to (1.6+/-0.3) times 10^{14} Msun from its measured X-ray luminosity. This ranks the cluster among intermediate mass system. In the case of XMMU J1532.2-0836 we find the X-ray detection to be coincident with a dynamically bound system of galaxies at z=1.358. Optical spectroscopy reveals the presence of a central active galactic nucleus, which can be a dominant source of the detected X-ray emission from this system. We provide upper limits of X-ray parameters for the system and discuss cluster identification challenges in the high-redshift low-mass cluster regime. A third, intermediate redshift (z=0.647) cluster, XMMU J0302.1-0000, is serendipitously detected in the same field as XMMU J0302.2-0001. We provide its analysis as well.Comment: Accepted to A&A, 13/04/2011. 15 pages, 18 figures, 5 tables, 2 appendice

Ancient oral microbiomes support gradual Neolithic dietary shifts towards agriculture

The human microbiome has recently become a valuable source of information about host life and health. To date little is known about how it may have evolved during key phases along our history, such as the Neolithic transition towards agriculture. Here, we shed light on the evolution experienced by the oral microbiome during this transition, comparing Palaeolithic hunter-gatherers with Neolithic and Copper Age farmers that populated a same restricted area in Italy. We integrate the analysis of 76 dental calculus oral microbiomes with the dietary information derived from the identification of embedded plant remains. We detect a stronger deviation from the hunter-gatherer microbiome composition in the last part of the Neolithic, while to a lesser extent in the early phases of the transition. Our findings demonstrate that the introduction of agriculture affected host microbiome, supporting the hypothesis of a gradual transition within the investigated populations

Multipronged dental analyses reveal dietary differences in last foragers and first farmers at Grotta Continenza, central Italy (15,500–7000 BP)

This paper provides results from a suite of analyses made on human dental material from the Late Palaeolithic to Neolithic strata of the cave site of Grotta Continenza situated in the Fucino Basin of the Abruzzo region of central Italy. The available human remains from this site provide a unique possibility to study ways in which forager versus farmer lifeways affected human odonto-skeletal remains. The main aim of our study is to understand palaeodietary patterns and their changes over time as reflected in teeth. These analyses involve a review of metrics and oral pathologies, micro-fossils preserved in the mineralized dental plaque, macrowear, and buccal microwear. Our results suggest that these complementary approaches support the assumption about a critical change in dental conditions and status with the introduction of Neolithic foodstuff and habits. However, we warn that different methodologies applied here provide data at different scales of resolution for detecting such changes and a multipronged approach to the study of dental collections is needed for a more comprehensive and nuanced understanding of diachronic changes