E-learning e futuri studenti in mobilità internazionale. Riflessioni su aspetti e potenzialità di un corso di lingua italiana

The A1 online Italian course offered by the CLA (University Linguistic Centre) of the University of Perugia, is one of the results achieved during various research projects, which has contributed, on the one hand, to the internationalization of the Institution, on the other, to the enhancement of digital technologies providing future university mobility students with the opportunity to acquire linguistic-cultural knowledge, even before the beginning of their mobility exchange programme in Italy. The experience reported in this article reflects on an evolving work, describing its design phase – course structure, selection and creation of linguistic and didactic materials, tools available in the Moodle open-source learning platform – and its subsequent phases of course activation and verification. Throughout the entire project, we focused on two fundamental aspects: inspiring and maintaining student motivation in addition to constructing an assisted, and above all, interactive self-learning path

Human trophoblast differentiation: possible role for trophoblast cell surface antigen 2

Human trophoblast cell surface antigen 2 (Trop2) is a 40-kDa transmembrane glycoprotein, encoded by TACSTD2 gene and identified for the first time in human trophoblast and choriocarcinoma cell lines. Trop2 has a short intracytoplasmic tail essential for the control of several pathways that regulate cellular functions such as cell- cell adhesion, cell proliferation and mobility [1]. We analysed the expression of Trop2 in human normal placentas during gestation and in placentas complicated by preeclampsia (PE). Trop2 protein expression and miR125b1 were analysed by morphological and bio-molecular techniques. Trop2 increased during gestation, i.e. from first to third trimester of gestation while it was low expressed in placental tissues collected from patients with PE. Since PE is a pathology associated with placental hypoxia, we demonstrated that Trop2 is downregulated in hypoxic conditions by in vitro model. Our study suggests a possible involvement of Trop2 in maintaining trophoblast morphology and function during placental development in normal and PE conditions

miR125b1 and TROP2 in preeclampsia complicated by foetal growth restriction: a morphological and biomolecular study

Trophoblast cell surface antigen 2 (TROP2) is a transmembrane glycoprotein originally identified in human trophoblast cell lines and is highly expressed in a variety of epithelial cancers. The TROP2 gene was validated as a direct target of miR-125b1. The purpose of our study was: - to investigate the expression of TROP2 protein in normal placental tissues, in placentas affected by preeclampsia as well as in placentas with preeclampsia complicated by foetal growth restriction (IUGR); - to verify how miR-125b1 was involved in the regulation of TROP2 gene expression. TROP2 protein expression was assessed by immunohistochemistry and quantitative western blotting analyses while miR-125b1 expression was detected by quantitative real-time PCR. The studies were made in normal and pathologic placental tissues. Increasing expression of TROP2 was detected in physiological placental tissue, in according with the increasing gestational age. Probably, it means that TROP2 is related with the differentiation of the cytotrophoblast in syncytiotrophoblast, that occurs during the development of placenta. Moreover, miR-125b1 showed an unchanged expression during normal pregnancy. Higher expression of TROP2 protein was detected in placental tissues collected from patients with preeclampsia complicated by foetal growth restriction, compared with those from preeclampsia and gestational age-matched control samples. The miR-125b1 expression in samples from placentas affected by preeclampsia complicated by IUGR was detected higher than in normal placentas and in placentas affected by preeclampsia. These results suggest that miR-125b1 is not involved I the overproduction of the TROP2 mRNA although the high expression of the miRNA. Our study suggests a possible involvement of TROP2 in the differentiation of the syncytiotrophoblast from villous cytotrophoblast and a possible role of this protein in preeclampsia complicated by foetal growth restriction

Analysis of tight junctions in placentas affected by chorioamnionitis: in vivo and in vitro analysis

The human placenta and fetal membranes provide a barrier regulating the transfer of materials between the mother and the developing fetus throughout gestation. Chorioamnionitis is an important risk factor for preterm delivery that is associated with high perinatal morbidity and mortality. Chorioamnionitis is the term applied to infections of the placenta and membranes resulting in high concentrations of IL- 1beta, IL-6, IL-8 and TGF-beta in the amniotic fluid (D’Alquen et al., 2005). With progression of inflammation, immune cells penetrate blood vessels and infiltrate the umbilical cord, resulting in funisitis (Romero and Mazor, 1988). In normal conditions the two important physical entities in endothelial/epithelial paracellular clefts are adherens junctions and tight junctions. Tight junction governs the paracellular movement of water, solutes and immune cells, through the intercellular space creating a boundary between the apical and basolateral sides of cellular barriers (Gruenheid and Finlay, 2003). We have evaluated the localization of tight junctions studying the Zonula Occludens-1 (ZO-1) and Occludin expressions as well as the localization of adherent junctions, testing the expression of VE-cadherin and beta-catenin in placentas from normal gestations, from preterm idiopathic deliveries and from chorioamnionitis by immunohistochemistry. In addition, we have evaluated the mRNAs by real time PCR, the protein levels of these molecules by Western blot analysis in placental tissues, and to better clarify the action of some cytokines on occludin we performed in vitro analysis of HUVEC cultures. Our more striking result is the decrease of occludin expression in placentas from chorioamnionitis and an evident action of the cytokines on this molecule

Expression of Trop2 in bladder cancer is modulated by miR125b: in vivo and in vitro analyses

Human trophoblastic cell surface antigen 2 (Trop-2) is a 40-kDa transmembrane glycoprotein, first identified as a cell surface marker for human trophoblast cells (1). Elevated expression of Trop-2 has been shown in several types of epithelial cancers and correlated with tumour aggressive and poor prognosis (2-3). The first aim of this study was to evaluate the variation of the Trop-2 expression in normal urothelium and urothelial bladder cancer. The immunohistochemical results showed an increase of Trop-2 levels in bladder cancer tissues with the increase of the severity of the pathology. Recent data identified Trop-2 as a target for miR-125b suggesting a pos sible role of miR-125b in the modulation of Trop-2 protein expression (4). The second aim was to verify if Trop-2 could be a target for miR-125b in bladder cells and to evaluate the possible role of miR-125b in the modulation of Trop-2 protein expression in normal bladder as well as in urothelial bladder cancer. In vitro we showed a contribution of miR-125b in deregulation of Trop-2 protein expression in a bladder cell line and we found that the expression of miR-125b was inversely correlated with the expression of Trop-2 protein on a cohort of bladder cancer tissues. We concluded to investigate in a larger population the use of Trop-2 and/or miR-125b as potential diagnostic markers in urothelial bladder cancer

HtrA1 in differentiation and growth of human placental tissues

HtrA1 is a secreted multidomain protein with serine protease activity. We used immunohistochemistry, western blotting, real time PCR and ELISA techniques to analyse the role of HtrA1 in normal and pathological development of human placental villous trees. In addition, we evaluated the alterations of maternal plasma HtrA1 level in preeclampsia (PE) complicated by intrauterine growth restriction (IUGR). HtrA1 is expressed in the mesenchymal villi which are considered the basis of growth and differentiation of the villous trees and in the villous stroma directly opposed to cell islands and cell columns in first trimester placentas. In addition, the villous trophoblast, the syncytial knots and the foetal vessels are stained for HtrA1 in first as well as third trimester placentas [1]. When the placenta escapes the normal differentiation and growth control mechanisms, which are present during normal pregnancy, it may develop gestational diseases, such as trophoblastic disease as well as PE and IUGR [1,2]. The most striking finding of our investigation is the decrease of this protease in placental tissues with increasing severity of gestational diseases and the increase of HtrA1 in maternal plasma of PE complicated by IUGR [3]. Based on these data HtrA1 could be considered as a possible marker of an occurring IUGR in preeclamptic women

Trop-2 expression and regulation in urothelial bladder cancer

Human trophoblastic cell surface antigen 2 (Trop-2) è una glicoproteina transmembrana di 40-kDa, identificata per la prima volta nelle cellule di trofoblasto umano. Trop-2 presenta un’elevata espressione in molti tipi di tumore epiteliale. Il primo scopo di questo lavoro è stato quello di valutare la variazione dell’espressione di Trop-2 nell’urotelio normale e nel cancro uroteliale della vescica. I risultati immunoistochimici hanno mostrato un aumento dei livelli di espressione di Trop-2 nei tessuti di cancro vescicale correlato con l’aumento della gravità della patologia. Dati recenti hanno identificato la proteina Trop-2 come target per il miR-125b suggerendo un possibile ruolo di questo miRNA nella modulazione dell’espressione di Trop-2. Il secondo obiettivo del lavoro è stato quello di verificare se Trop-2 poteva agire da target per il miR-125b a livello delle cellule vescicali e di valutare un possibile ruolo del miRNA nella modulazione dell’espressione di Trop-2 nei tessuti vescicali fisiologici cosi come in quelli tumorali. In vitro, abbiamo mostrato come il miR-125b contribuisca alla disregolazione di Trop-2 nelle cellule epiteliali di vescica. Abbiamo inoltre visto come su un gruppo di tessuti di cancro uroteliale della vescica l’espressione del miR-125b sia inversamente correlata all’espressione di Trop-2. In conclusione, suggeriamo di valutare su dimensione campionaria più ampia l’uso di una o di entrambe le molecole come potenziali marker diagnostici nel carcinoma uroteliale della vescica.Human trophoblastic cell surface antigen 2 (Trop-2) is a 40-kDa transmembrane glycoprotein, first identified as a cell surface marker for human trophoblast cells. Elevated expression of Trop-2 has been documented in several types of epithelial cancers. The first aim of this study was to evaluate the variation of the Trop-2 expression in normal urothelium and urothelial bladder cancer. The immunohistochemical results showed an increase of Trop-2 levels in bladder cancer tissues with the increase of the severity of the pathology. Recent data identified Trop-2 as a target for miR-125b suggesting a possible role of miR-125b in the modulation of Trop-2 protein expression. The second aim was to verify if Trop-2 could be a target for miR-125b in bladder cells and to evaluate the possible role of miR-125b in the modulation of Trop-2 protein expression in normal bladder as well as in urothelial bladder cancer. In vitro we showed a contribution of miR-125b in deregulation of Trop-2 protein expression in a bladder cell line and we found that the expression of miR-125b was inversely correlated with the expression of Trop-2 protein on a cohort of bladder cancer tissues. We concluded to investigate in a larger population the use of Trop-2 and/or miR-125b as potential diagnostic markers in urothelial bladder cancer

Analysis of cell-cell junctions in human amnion and chorionic plate affected by chorioamnionitis

Chorioamnionitis is an acute inflammatory reaction associated with the premature rupture of the fetal membranes. It is caused mainly by invasion of bacteria from the vaginal tract that can penetrate the intact membranes and invade the amnion cavity and the decidua. Tight junctions (TJ) and adherent junctions (AJ) are intercellular junctions crucial for epithelia adhesion and permeability regulation in a wide variety of tissues and organs. Our aim is to investigate if TJ and AJ molecules are involved in human chorioamnionitis. We studied the protein expression (by immunohistochemistry and western blotting) and the mRNA levels (by RT-PCR) of some junction proteins such as Zonula Occludens-1 (ZO-1), occludin, VE-cadherin and βcatenin in fetal membranes from women with chorioamnionitis compared to those membranes derived from idiopathic pregnancies. Western blotting and immunohistochemical data established that occludin expression was decreased in amnion with chorioamnionitis compared to amnion from idiopathic pregnancies. Samples tested for ZO-1, VEcadherin and β-catenin (proteins and mRNAs) showed no differences between idiopathic and pathological membranes. One of the most relevant results is the decrease of occludin in membranes with chorioamnionitis. Since we have previously demonstrated that some cytokines, particularly elevated in the chorioamnionitis, cause the disruption of TJs in placental villi, we suggest that the decrease of occludin in amnion may be the first change that leads to the rupture of the amniotic membrane in this pathology