Endothelial-Mesenchymal Transition of Brain Endothelial Cells: Possible Role during Metastatic Extravasation

Cancer progression towards metastasis follows a defined sequence of events described as the metastatic cascade. For extravasation and transendothelial migration metastatic cells interact first with endothelial cells. Yet the role of endothelial cells during the process of metastasis formation and extravasation is still unclear, and the interaction between metastatic and endothelial cells during transendothelial migration is poorly understood. Since tumor cells are well known to express TGF-beta, and the compact endothelial layer undergoes a series of changes during metastatic extravasation (cell contact disruption, cytoskeletal reorganization, enhanced contractility), we hypothesized that an EndMT may be necessary for metastatic extravasation. We demonstrate that primary cultured rat brain endothelial cells (BEC) undergo EndMT upon TGF-beta 1 treatment, characterized by the loss of tight and adherens junction proteins, expression of fibronectin, beta 1-integrin, calponin and a-smooth muscle actin (SMA). B16/F10 cell line conditioned and activated medium (ACM) had similar effects: claudin-5 down-regulation, fibronectin and SMA expression. Inhibition of TGF-beta signaling during B16/F10 ACM stimulation using SB-431542 maintained claudin-5 levels and mitigated fibronectin and SMA expression. B16/F10 ACM stimulation of BECs led to phosphorylation of Smad2 and Smad3. SB-431542 prevented SMA up-regulation upon stimulation of BECs with A2058, MCF-7 and MDA-MB231 ACM as well. Moreover, B16/F10 ACM caused a reduction in trans-endothelial electrical resistance, enhanced the number of melanoma cells adhering to and transmigrating through the endothelial layer, in a TGF-beta-dependent manner. These effects were not confined to BECs: HUVECs showed TGF-beta-dependent SMA expression when stimulated with breast cancer cell line ACM. Our results indicate that an EndMT may be necessary for metastatic transendothelial migration, and this transition may be one of the potential mechanisms occurring during the complex phenomenon known as metastatic extravasation

Conceptual design of the FAST load assembly

Fusion advanced studies torus (FAST) is a proposal for a satellite facility which can contribute the rapid exploitation of ITER and prepare ITER and DEMO regimes of operation, as well as exploiting innovative DEMO technology. FAST is a compact (R0 = 1.82 m, a = 0.64 m, triangularity [delta] = 0.4) machine able to investigate non-linear dynamics effects of alpha particle behaviours in burning plasmas [1], [2] and [5]. The project is based on a dominant 30 MW of ion cyclotron resonance heating (ICRH), 6 MW of lower hybrid (LH) and 4 MW of electron cyclotron resonance heating (ECRH). FAST operates at a wide range [3] and [4] of parameters, e.g., in high performance H-mode (BT up to 8.5 T; IP up to 8 MA) as well as in advanced Tokamak operation (IP = 3 MA), and full non-inductive current scenario (IP = 2 MA). Helium gas at 30 K is used for cooling the resistive copper magnets [6]. That allows for a pulse duration up to 170 s. To limit the TF magnet ripple ferromagnetic insert have been introduced inside the vacuum vessel (VV). Ports have been designed to also accommodate up to 10 MW of negative neutral beam injection (NNBI). Tungsten (W) or liquid lithium (L-Li) have been chosen as the divertor plates material, and argon or neon as the injected impurities to mitigate the thermal loads

Tie2 Expressing Monocytes in the Spleen of Patients with Primary Myelofibrosis

<div><p>Primary myelofibrosis (PMF) is a Philadelphia-negative (Ph−) myeloproliferative disorder, showing abnormal CD34⁺ progenitor cell trafficking, splenomegaly, marrow fibrosis leading to extensive extramedullary haematopoiesis, and abnormal neoangiogenesis in either the bone marrow or the spleen. Monocytes expressing the angiopoietin-2 receptor (Tie2) have been shown to support abnormal angiogenic processes in solid tumors through a paracrine action that takes place in proximity to the vessels. In this study we investigated the frequency of Tie2 expressing monocytes in the spleen tissue samples of patients with PMF, and healthy subjects (CTRLs), and evaluated their possible role in favouring spleen angiogenesis. We show by confocal microscopy that in the spleen tissue of patients with PMF, but not of CTRLs, the most of the CD14⁺ cells are Tie2⁺ and are close to vessels; by flow cytometry, we found that Tie2 expressing monocytes were Tie2⁺CD14^lowCD16^brightCDL62⁻CCR2⁻ (TEMs) and their frequency was higher (p = 0.008) in spleen tissue-derived mononuclear cells (MNCs) of patients with PMF than in spleen tissue-derived MNCs from CTRLs undergoing splenectomy for abdominal trauma. By <i>in vitro</i> angiogenesis assay we evidenced that conditioned medium of immunomagnetically selected spleen tissue derived CD14⁺ cells of patients with PMF induced a denser tube like net than that of CTRLs; in addition, CD14⁺Tie2⁺ cells sorted from spleen tissue derived single cell suspension of patients with PMF show a higher expression of genes involved in angiogenesis than that found in CTRLs. Our results document the enrichment of Tie2⁺ monocytes expressing angiogenic genes in the spleen of patients with PMF, suggesting a role for these cells in starting/maintaining the pathological angiogenesis in this organ.</p></div