P. Di Marco

Parthenolide generates reactive oxygen species and autophagy in MDA-MB231 cells. A soluble parthenolide analogue inhibits tumour growth and metastasis in a xenograft model of breast cancer

Triple-negative breast cancers (TNBCs) are clinically aggressive forms associated with a poor prognosis. We evaluated the cytotoxic effect exerted on triple-negative MDA-MB231 breast cancer cells both by parthenolide and its soluble analogue dimethylamino parthenolide (DMAPT) and explored the underlying molecular mechanism. The drugs induced a dose- and time-dependent decrement in cell viability, which was not prevented by the caspase inhibitor z-VAD-fmk. In particular in the first hours of treatment (1–3 h), parthenolide and DMAPT strongly stimulated reactive oxygen species (ROS) generation. The drugs induced production of superoxide anion by activating NADPH oxidase. ROS generation caused depletion of thiol groups and glutathione, activation of c-Jun N-terminal kinase (JNK) and downregulation of nuclear factor kB (NF-kB). During this first phase, parthenolide and DMAPT also stimulated autophagic process, as suggested by the enhanced expression of beclin-1, the conversion of microtubule-associated protein light chain 3-I (LC3-I) to LC3-II and the increase in the number of cells positive to monodansylcadaverine. Finally, the drugs increased RIP-1 expression. This effect was accompanied by a decrement of pro-caspase 8, while its cleaved form was not detected and the expression of c-FLIPS markedly increased. Prolonging the treatment (5–20 h) ROS generation favoured dissipation of mitochondrial membrane potential and the appearance of necrotic events, as suggested by the increased number of cells positive to propidium iodide staining. The administration of DMAPT in nude mice bearing xenografts of MDA-MB231 cells resulted in a significant inhibition of tumour growth, an increment of animal survival and a marked reduction of the lung area invaded by metastasis. Immunohistochemistry data revealed that treatment with DMAPT reduced the levels of NF-kB, metalloproteinase-2 and -9 and vascular endothelial growth factor, while induced upregulation of phosphorylated JNK. Taken together, our data suggest a possible use of parthenolide for the treatment of TNBCs.

Differentiation and characterization of rat adipose tissue mesenchymal stem cells into endothelial-like cells

In this study, mesenchymal stem cells were isolated from rat adipose tissue (AD‐MSCs) to characterize and differentiate them into endothelial‐like cells. AD‐MSCs were isolated by mechanical and enzymatic treatments, and their identity was verified by colony‐forming units (CFU) test and by differentiation into cells of mesodermal lineages. The endothelial differentiation was induced by plating another aliquot of cells in EGM‐2 medium, enriched with specific endothelial growth factors. Five subcultures were performed. The expression of stemness genes (OCT4, SOX2 and NANOG) was investigated. The presence of CD90 and the absence of the CD45 were evaluated by flow cytometry. The endothelial‐like cells were characterized by the evaluation of morphological changes and gene expression analysis for endothelial markers (CD31, CD144, CD146). Characterization of AD‐MSCs showed their ability to form clones, to differentiate in vitro and the OCT‐4, SOX‐2, NANOG genes expression. Immunophenotypic characterization showed the CD90 presence and the CD45 absence. The endothelial‐like cells showed morphological changes, the expression of CD31, CD144, CD146 genes and the presence of CD31 membrane receptor. Matrigel assay showed their ability to form network and vessels‐like structures. This study lays the foundations for future evaluation of the potential AD‐MSCs pro‐angiogenic and therapeutic role.

Obtaining Mesenchymal Stem Cells From Adipose Tissue Of Murin Origin: Experimental Study

1 Phd School in Surgical Biotechnology and Regenerative Medicine. School of Medicine – School of Biotechnology, University of Palermo, Italy. 2 DICHIRONS Department, School of Medicine. University of Palermo. Italy. 3 Istituto Zooprofilattico Sperimentale della Sicilia A.Mirri, Palermo, Italy. 4 AUOP “P. Giaccone”, Universitary Hospital, Palermo, Italy. 5 School of Biotechnology, University of Palermo, Italy. 6 Euromediterranean Institute of Science and Technology (IEMEST), Palermo, Italy.

West Nile Disease (WND) in Sicily

BACKGROUND WND is an emerging vector-borne zoonosis caused by an RNA virus included in the Japanese encephalitis group within the Flavivirus genus (3, 5). The virus is reported in several countries of the Mediterranean Basin (Fig. 2) and is maintained in nature by cycling through birds and mosquitoes (Fig. 1). In Italy, after the first appearance in Tuscany in 1998 (1), West Nile Virus (WNV) has been continuously circulating since 2008. A National Surveillance Program consisting of monitoring horses, “sentinel poultries”, wild bird mortality and mosquitoes is active in the entire national territory since 2002 (4). Authors describe the outbreaks which occurred in Sicily in 2010 and 2011 (Fig. 3). Case study: Between September-October 2010, seven horses from the western part of the island, in the Trapani province, showed neurological symptoms (2). In October 2011, another two horses suffering from neurological disorders, were observed around Messina, in the eastern part of the island, and near Palermo, in the north west of Sicily.