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Background Drug toxicity currently represents the main challenge of tumour chemotherapy. Our group recently developed a new method for drug delivery inspired by the 'Trojan Horse' concept. Human mesenchymal stem cells (hMSCs) have been shown to play the role of new 'horses' in delivering anti-tumour agents, without involving any genetic manipulation. As human stromal dermal fibroblasts (hSDFs) represent an interesting alternative to hMSCs, being easy to isolate, they could be an ideal candidate for this kind of procedure. Aim To investigate whether hSDFs can take up and deliver paclitaxel (PTX) in sufficient concentrations to inhibit a very aggressive melanoma tumour (IgR39) in vitro. Methods hSDFs were primed with high doses of PTX, and then the effect of drug delivery on IgR39 melanoma proliferation in vitro was evaluated using several assays (antiproliferation, transwell cocultures, rosette assays and colony growth assays). Furthermore, the cell cycle and PTX uptake/release mechanism of hSDFs were studied both under both normal and hypoxic conditions. Results hSDFs incorporated PTX and then released it with unaffected pharmacological activity, inhibiting human IgR39 melanoma growth in vitro. The hypoxic conditions did not induce changes in cell cycle pattern and the uptake-release mechanism with PTX was not affected. Conclusions hSDFs can be used as a Trojan horse, as the released drug was functionally active. These results indicated that these cells could be used for clinical treatment as the drug was released into the cellular environment and the primed cells underwent apoptosis.

We recently developed a method for the isolation and purification of tumour-derived endothelium. In this study the phenotypic and functional properties of human tumour-derived microvascular endothelial cells (TdMEC) were examined. Endothelium obtained from human adrenal gland specimens (HAMEC) was used as a reference microvascular endothelial cell population. TdMEC formed a confluent monolayer with the typical morphological appearance of endothelium and were positive for endothelial markers such as Ulex-1 lectin, CD31 antigen, von Willebrand Factor and VE-cadherin. The addition of acidic Fibroblast Growth Factor (aFGF), basic FGF (bFGF) or Vascular Endothelial Growth Factor (VEGF) substantially improved proliferation of TdMEC; and kidney carcinoma derived endothelial cells were more responsive to FGFs, whereas glioblastoma derived endothelial cells greatly responded to VEGF. TdMEC expressed high levels of the VEGF receptors, KDR/flk-1 and Flt-1, as shown by northern blot analysis. TdMEC expressed the adhesion molecules ICAM-1, VCAM-1 and E-selectin that could be further increased by exposing TdMEC culture to interleukin-1. All the TdMEC expressed interleukin-8 mRNA. These findings show that TdMEC in vitro maintain several of the features described for microvasculature. Thus, TdMEC represent a useful tool to study markers for tumor vasculature

L'anestesia al linguaggio simbolico delle cose e delle esperienze nel mondo contemporaneo è aumentata enormemente. Occorre esercitare, attraverso l'osservazione accurata e paziente di opere simboliche, questa facoltà vitale, per l'uomo e il suo ambiente. In questo senso il ruolo dell'arte, sotto il profilo formativo, è nevralgico.

Mesenchymal stromal cells (MSCs) have been proposed for delivering anticancer agents because of their ability to home in on tumor microenvironment. We found that MSCs can acquire strong anti-tumor activity after priming with Paclitaxel (PTX) through their capacity to uptake and then release the drug. Because MSCs secrete a high amount of membrane microvesicles (MVs), we here investigated the role of MVs in the releasing mechanism of PTX. The murine SR4987 line was used as MSC model. The release of PTX from SR4987 in the conditioned medium (CM) was checked by HPLC and the anti-tumor activity of both CM and MVs was tested on the human pancreatic cell line CFPAC-1. MVs were isolated by ultracentrifugation, analyzed by transmission (TEM) and scanning electron microscopy (SEM), and the presence of PTX by the Fourier transformed infrared (FTIR) microspectroscopy. SR4987 loaded with PTX (SR4987PTX) secreted a significant amount of PTX and their CM possessed strong anti-proliferative activity on CFPAC-1. At TEM and SEM, SR4987PTX showed an increased number of "vacuole-like" structures and shed a relevant number of MVs, but did not differ from untreated SR4987. However, SR4987PTX-derived-MVs (SR4987PTX-MVs) demonstrated a strong anti proliferative activity on CFPAC-1. FTIR analysis of SR4987PTX-MVs showed the presence of an absorption spectrum in the corresponding regions of the PTX marker, absent in MVs from SR4987. Our work is the first demonstration that MSCs are able to package and deliver active drugs through their MVs, suggesting the possibility of using MSCs as a factory to develop drugs with a higher cell-target specificity. © 2014 Elsevier B.V.

Cyclin-dependent kinase-5 (Cdk5) is over-expressed in both neurons and microvessels in hypoxic regions of stroke tissue and has a significant pathological role following hyper-phosphorylation leading to calpain-induced cell death. Here, we have identified a critical role of Cdk5 in cytoskeleton/focal dynamics, wherein its activator, p35, redistributes along actin microfilaments of spreading cells co-localising with p(Tyr15)Cdk5, talin/integrin beta-1 at the lamellipodia in polarising cells. Cdk5 inhibition (roscovitine) resulted in actin-cytoskeleton disorganisation, prevention of protein co-localization and inhibition of movement. Cells expressing Cdk5 (D144N) kinase mutant, were unable to spread, migrate and form tube-like structures or sprouts, while Cdk5 wild-type over-expression showed enhanced motility and angiogenesis in vitro, which was maintained during hypoxia. Gene microarray studies demonstrated myocyte enhancer factor (MEF2C) as a substrate for Cdk5-mediated angiogenesis in vitro. MEF2C showed nuclear co-immunoprecipitation with Cdk5 and almost complete inhibition of differentiation and sprout formation following siRNA knock-down. In hypoxia, insertion of Cdk5/p25-inhibitory peptide (CIP) vector preserved and enhanced in vitro angiogenesis. These results demonstrate the existence of critical and complementary signalling pathways through Cdk5 and p35, and through which coordination is a required factor for successful angiogenesis in sustained hypoxic condition. © 2013 Bosutti et al.

Mesenchymal stem cells (MSCs) were first isolated more than 50 years ago from the bone marrow. Currently MSCs may also be isolated from several alternative sources and they have been used in more than a hundred clinical trials worldwide to treat a wide variety of diseases. The MSCs mechanism of action is undefined and currently under investigation. For in vivo purposes MSCs must be produced in compliance with good manufacturing practices and this has stimulated research on MSCs characterization and safety. The objective of this review is to describe recent developments regarding MSCs properties, physiological effects, delivery, clinical applications and possible side effects.

Five studies document the validity of a new 8-item scale designed to measure positivity, defined as the tendency to view life and experiences with a positive outlook. In the first study (N = 372), the psychometric properties of Positivity Scale (P Scale) were examined in accordance with classical test theory using a large number of college participants. In Study 2, the unidimensionality of the P Scale was corroborated with confirmatory factor analysis in 2 independent samples (N1 = 322; N2 = 457). In Study 3, P Scale invariance across sexes and its relations with self-esteem, life satisfaction, optimism, positive negative affect, depression, and the Big Five provided further evidence of the internal and construct validity of the new measure in a large community sample (N = 3,589). In Study 4, test–retest reliability of the P Scale was found in a sample of college students (N = 262) who were readministered the scale after 5 weeks. In Study 5, measurement invariance and construct validity of P Scale were further supported across samples in different countries and cultures, including Italy (N = 689), the United States (N = 1,187), Japan (N = 281), and Spain (N = 302). Psychometric findings across diverse cultural context attest to the robustness of the P Scale and to positivity as a basic disposition

Aim – Literature documents that the judgments people hold about themselves, their life, and their future are important ingredients of their psychological functioning and well-being, and are commonly related to each other. Methods – We used confirmatory factor analysis (CFA) under the classical test theory, regression analysis, and a cross national design. Results – This study provides new findings attesting to the hypothesis that evaluations about oneself, one’s life, and one’s future rest on a common mode of viewing experiences which we named “Positive Orientation”. Conclusions – Results from an Italian and a Canadian study are presented, attesting to a latent dimension that lies at the core of positive evaluations and that corroborates the utility of the new construct as a critical component of individuals’ well-functioning.

This study aims to explore the utility of resilient, overcontrolled and undercontrolled personality prototypes in discriminating 735 elderly Italian adults with regards to their well-being, quality of interpersonal relationships, and leisure activities. Prototype membership, corresponding to the three types, was identified through cluster analysis of Big Five self-ratings. The three prototypes clearly differed in terms of their life and health satisfaction, positive affectivity, interpersonal trust, civic and social engagement and leisure activities. Resilients showed the most positive profile; undercontrollers mistrusted their family members, whereas overcontrollers reported particularly low satisfaction and a scarce involvement in social and recreation activities. © 2009 Elsevier Ltd. All rights reserved.

The present study examined the longitudinal relations between individuals' prosociality and their self-efficacy beliefs in regard to emotional regulation and responding empathically to others' needs. The participants were 244 females and 222 males with a mean age of 17 years (SD=1.5) at T1, 19 years (SD=1.4) at T2, and 21 years (SD=1.6) at T3. The findings corroborated the posited paths of relations assigning empathic self-efficacy a major role in predicting the level of individuals' prosociality. Empathic self-efficacy beliefs mediated the relations of regulative emotional self-efficacy beliefs to prosocial tendencies such as caring, sharing, helping, and empathic concern toward others. The posited conceptual model accounted for a significant portion of variance in prosociality and has implications for interventions designed to promote and sustain prosociality. © 2009, Wiley Periodicals, Inc.

Certain personality characteristics such as self-esteem, life satisfaction, and optimism are fundamental components of positive mental health status and well-being. There is consistent evidence that these traits tend to be substantially correlated in individuals. However, no previous studies have investigated the origin of such correlation. This research used the twin method to unravel the genetic and environmental architecture of self-esteem, life satisfaction, and optimism, along with their mutual interplay. The sample was derived from the population-based Italian Twin Register, and included 428 twin pairs, aged 23-24 years. Multivariate genetic modeling showed that genes influencing self-esteem, life satisfaction, and optimism are largely overlapping. Furthermore, results indicated that the environmental components of the traits may overlap only modestly, and suggested that a sizeable amount of variance in the traits may be explained by environmental effects specific to each of them. © 2009 Springer Science+Business Media, LLC.