In vitro study of the effects of endothelin-1: relevance to the pathogenesis of pulmonary arterial hypertension in systemic sclerosis.

Background: Pulmonary arterial hypertension is a devastating disease with an estimated patients\u2019 survival of 68% at 1 year, 48% at 3 years and 34% at 5 years from diagnosis. The role of endothelin-1 has been studied in different conditions including systemic sclerosis. In systemic sclerosis patients, circulating endothelin- 1 is present at high levels, suggesting its involvement in the development of fibrotic and vascular damage. Indeed endothelin-1 is a potent pro-fibrotic peptide and inhibitors of endothelin-1 receptors (ETA and ETB) may exert an anti-fibrotic effect. The balance of endothelin-1 effects on smooth muscle cells and on endothelial cells is responsible for the homeostasis of vascular tone. Another interesting effect of endothelin-1 is the induction of aldosterone in endothelial cells that eventually results in the production of reactive oxygen species. Aldosterone is a key mediator of the pulmonary vascular injury response to hypoxia: mineralocorticoid receptor activation by aldosterone is involved in the remodeling/fibrosis response to hypoxia in endothelial cells in vitro and aldosterone antagonism attenuates these events. Methods: Reverse transcriptase-PCR, western blot and cytofluorimetric analyses were performed to evaluate the expression of the two isoforms of endothelin-1 receptors and mineralocorticoid receptor. Activation of fibroblast was evaluated measuring the amount of collagen-1, TGF-\u3b2 and PDGF with ELISA kits. The detection of intracellular oxidative stress formation was measured with flow cytometry using fluorescence of CM-H2DCFDA after long or short incubations with endothelin-1 and its receptors inhibitors or with aldosterone and its antagonist. Results: Both endothelin-1 receptors are present in commercially available human pulmonary fibroblasts and normal human dermal fibroblasts, human dermal fibroblasts from healthy donors and scleroderma human dermal fibroblasts. Mineralocorticoid receptor expression was detected in human pulmonary fibroblasts, scleroderma human dermal fibroblasts and human pulmonary artery endothelial cells while in normal human dermal fibroblasts and human dermal fibroblasts from healthy donors the amount of this receptor was lower than the other cells analyzed. In human pulmonary fibroblasts, scleroderma human dermal fibroblasts and human pulmonary artery endothelial cells, endothelin-1 stimulates the production of reactive oxygen species after a long incubation. In addition, the production of oxygen radicals is inhibited by the use of ETB antagonist. Interestingly, aldosterone stimulates the production of reactive oxygen species after a short incubation in human pulmonary fibroblasts, scleroderma human dermal fibroblasts and human pulmonary artery endothelial cells, but not in normal human dermal fibroblasts and human dermal fibroblasts from healthy donors. The same results were obtained by human pulmonary fibroblasts, scleroderma human dermal fibroblasts and human pulmonary artery endothelial cells stimulated with endothelin-1 for long time with mineralocorticoid receptor inhibitor. Conclusions: These results suggest that endothelin-1 activates fibroblasts and the production of aldosterone, the true responsible for oxidative stress. The inhibition of reactive oxygen species production using the ETB antagonist indicates that the ETB receptor is implicated in this process. Moreover we found differences between fibroblasts from healthy donors and from scleroderma patients in oxidative stress production after endothelin-1 stimulation. Finally we found differences in mineralocorticoid receptor presence in the same cell types

Linked Data approach for selection process automation in Systematic Reviews

Background: a systematic review identifies, evaluates and synthesizes the available literature on a given topic using scientific and repeatable methodologies. The significant workload required and the subjectivity bias could affect results. Aim: semi-automate the selection process to reduce the amount of manual work needed and the consequent subjectivity bias. Method: extend and enrich the selection of primary studies using the existing technologies in the field of Linked Data and text mining. We define formally the selection process and we also develop a prototype that implements it. Finally, we conduct a case study that simulates the selection process of a systematic literature published in literature. Results: the process presented in this paper could reduce the work load of 20% with respect to the work load needed in the fully manually selection, with a recall of 100%. Conclusions: the extraction of knowledge from scientific studies through Linked Data and text mining techniques could be used in the selection phase of the systematic review process to reduce the work load and subjectivity bia

O8D.8 Severity of carpal tunnel syndrome and manual work: findings from a case-control study

ObjectiveCarpal tunnel syndrome (CTS) is a socially relevant condition. This case-control study aims to investigate the association between CTS severity and manual work considering personal anthropometric risk factors as well.MethodsWe consecutively enrolled one CTS case for two controls (subjects without clinical and electrophysiological CTS signs) regardless of age and gender who were admitted to the same three outpatient electromyography labs.CTS cases were grouped in three classes of progressive clinical and electrophysiological severity according to two validated five-stage scales. Anthropometric measures and occupational history were collected. Job titles were coded according to the International Standard Classification of Occupations (ISCO 88) by two occupational physicians who were blind to case/control status. Job titles were grouped in two main occupational categories: manual workers and non-manual workers.To assess the association between CTS severity and manual work, ordered logistic regression models (adjusted for age, sex, wrist-palm ratio and waist-stature ratio) were performed. Odds ratios (OR) and 95% confidence intervals (95% CI) were calculated.ResultsThis case-control study enrolled 370 cases and 747 controls. After the exclusion of retired subjects, subjects older than 65 years and subjects with no information about occupational history, we included 183 cases and 445 controls in the main analysis.For manual workers with respect to non-manual workers, the OR for the electrophysiological severity scale was 2.4 (95%CI 1.5–3.7). Regarding the clinical severity scale, the OR for manual workers compared to non-manual workers were 2.3 (95%CI 1.5–3.7).ConclusionThis study confirms that manual work is an important risk factor for CTS. The association between manual work and CTS severity tends to increase from mild to severe stage of both electrophysiological and clinical scale

Endothelin Receptors Expressed by Immune Cells Are Involved in Modulation of Inflammation and in Fibrosis: Relevance to the Pathogenesis of Systemic Sclerosis

open13noEndothelin-1 (ET-1) plays a pivotal role in vasoconstriction, fibrosis, and inflammation, the key features of systemic sclerosis (SSc). ET-1 receptors (ETA and ETB) are expressed on endothelial cells, smooth muscle cells, and fibroblasts, but their presence on immune cells has not been deeply investigated so far. Endothelin receptors antagonists such as bosentan have beneficial effects on vasoconstriction and fibrosis, but less is known about their potential anti-inflammatory effects. We studied the expression of ET-1 receptors on immune cells (T and B lymphocytes, monocytes, and neutrophils) and the link between ET-1 and inflammation in patients with SSc. We show here that ET-1 exerts a proinflammatory effect in CD4+ T cells, since it induces an increased IFN-γ production; preincubation with antagonists of both receptors reduces IFN-γ production. Moreover, following ET-1 stimulation, neutrophils produce proinflammatory mediators, thus amplifying the effects of activated CD4+ T cells. Our data indicate that ET-1 system is involved in the pathogenesis of inflammation and fibrosis typical of SSc, through the activation of T lymphocytes and neutrophils and the consequent release of proinflammatory and profibrotic cytokines. These findings suggest that dual ET-1 receptors antagonist therapy, besides its effect on vasculopathy, has a profound impact on the immune system favouring antiinflammatory and antifibrogenic effects.openElisa, Tinazzi; Antonio, Puccetti; Giuseppe, Patuzzo; Alessandro, Barbieri; Giuseppe, Argentino; Federico, Confente; Marzia, Dolcino; Ruggero, Beri; Giacomo, Marchi; Andrea, Ottria; Daniela, Righetti; Mariaelisa, Rampudda; Claudio, LunardiTinazzi, Elisa; Puccetti, Antonio; Patuzzo, Giuseppe; Barbieri, Alessandro; Argentino, Giuseppe; Confente, Federico; Dolcino, Marzia; Beri, Ruggero; Marchi, Giacomo; Ottria, Andrea; Righetti, Daniela; Rampudda, Mariaelisa; Lunardi, Claudi

Hidden Comorbidities in Asthma: A Perspective for a Personalized Approach

: Bronchial asthma is the most frequent inflammatory non-communicable condition affecting the airways worldwide. It is commonly associated with concomitant conditions, which substantially contribute to its burden, whether they involve the lung or other districts. The present review aims at providing an overview of the recent acquisitions in terms of asthma concomitant systemic conditions, besides the commonly known respiratory comorbidities. The most recent research has highlighted a number of pathobiological interactions between asthma and other organs in the view of a shared immunological background underling different diseases. A bi-univocal relationship between asthma and common conditions, including cardiovascular, metabolic or neurodegenerative diseases, as well as rare disorders such as sickle cell disease, α1-Antitrypsin deficiency and immunologic conditions with hyper-eosinophilia, should be considered and explored, in terms of diagnostic work-up and long-term assessment of asthma patients. The relevance of that acquisition is of utmost importance in the management of asthma patients and paves the way to a new approach in the light of a personalized medicine perspective, besides targeted therapies

Profibrotic Effects of Endothelin-1 on Fibroblasts Are Mediated by Aldosterone in Vitro: Relevance to the Pathogenesis and Therapy of Systemic Sclerosis and Pulmonary Arterial Hypertension

Endothelin-1 (ET-1) is a vasoactive and profibrotic peptide that plays a pivotal role in diseases such as systemic sclerosis (SSc) and pulmonary arterial hypertension (PAH), by inducing fibrosis and vascular remodeling. Such effects may be sustained by the induction of aldosterone production and reactive oxygen species (ROS). We have used fibroblasts obtained from skin of healthy donors and SSc patients and commercial fibroblasts from lung to evaluate whether ET-1 is able to stimulate ROS production directly or indirectly through aldosterone induction. We found that ET-1 receptors are present in all types of fibroblasts analyzed, whereas the expression of mineralocorticoid receptor (MCR) is lower in dermal fibroblasts from healthy donors (HDFs) compared to fibroblasts derived from lung (HPFs) or from skin of SSc patients (SScHDFs). ET-1 induces ROS production in HDFs and SScHDFs after 24 h of incubation involving its receptor B (ETB), whereas aldosterone exerts its effects after 40 min of incubation. Moreover, ROS production was inhibited by the pre-incubation of cells with MCR inhibitor. Our results indicate that ET-1 induces ROS indirectly through aldosterone production suggesting that aldosterone may play a pivotal role in the pathogenesis of SSc and PAH

IN VITRO STUDY OF THE EFFECTS OF ENDOTELIN-1: RELEVANCE TO THE PATHOGENESIS OF PULMONARY ARTERIAL HYPERTENSION IN SYSTEMIC SCLEROSIS

Introduzione: L'ipertensione arteriosa polmonare è una patologia grave con una sopravvivenza dei pazienti stimata del 68% a 1 anno, 48% a 3 anni e il 34% a 5 anni dalla diagnosi. Il ruolo dell’endotelina-1 è stato studiato in diverse condizioni patologiche tra le quali la sclerosi sistemica; nei pazienti affetti da sclerosi sistemica, l’endotelina-1 circolante è presente ad alto titolo suggerendo il suo coinvolgimento nello sviluppo del danno fibrotico e vascolare. L’endotelina-1 è, infatti, un peptide con attività pro-fibrotica, pertanto gli inibitori dei recettori endotelinici (ETA e ETB) possono esercitare un effetto anti-fibrotico. A livello vasale, inoltre, l'equilibrio degli effetti mediati dall’endotelina-1 sulle cellule muscolari lisce e sulle cellule endoteliali è responsabile dell'omeostasi del tono vascolare. Un altro interessante effetto dell’endotelina-1 sulle cellule endoteliali è l'induzione della produzione di aldosterone che, a sua volta, sembrerebbe incrementare i livelli di specie reattive dell'ossigeno. L'aldosterone è un mediatore chiave nel danno vascolare polmonare in risposta all’ipossia: l'attivazione del recettore per mineralcorticoidi dovuto all’aldosterone nelle cellule endoteliali in vitro è coinvolta nel rimodellamento vascolare e nella fibrosi in risposta all'ipossia e l’utilizzo di antagonisti aldosteronici attenua questi fenomeni.Metodi: Per valutare l'espressione delle due isoforme recettoriali dell’endotelina-1 e del recettore per mineralcorticoidi, sono state effettuate analisi di reverse transcriptase-PCR, western blot e citofluorimetria. L'attivazione dei fibroblasti è stata valutata misurando con kit ELISA la quantità di collagene-1, TGF-β e PDGF nel terreno di coltura. La rilevazione della formazione di stress ossidativo intracellulare è stata valutata con citometria a flusso misurando la fluorescenza emessa dalla CM-H2DCFDA dopo incubazioni lunghe o brevi con endotelina-1 e i suoi inibitori recettoriali o con l’aldosterone e il suo antagonista.Risultati: Entrambi i recettori per l’endotelina-1 sono presenti sulla superficie dei fibroblasti polmonari umani e dei fibroblasti dermici normali umani disponibili incommercio, ma anche su quella dei fibroblasti dermici ottenuti da donatori sani e da pazienti sclerodermici. L’espressione del recettore per mineralcorticoidi è stata rilevata nei fibroblasti polmonari umani, nei fibroblasti dermici sclerodermici e nelle cellule endoteliali polmonari umane mentre nei fibroblasti dermici normali umani e in quelli da donatori sani, la quantità di proteina recettoriale rilevata è stata più bassa che nelle altre cellule analizzate. Nei fibroblasti polmonari umani, in quelli dermici sclerodermici e nelle cellule endoteliali polmonari umane, l’endotelina-1 ha stimolato la produzione di specie reattive dell’ossigeno dopo incubazioni lunghe; tale produzione è stata inibita dall’utilizzo del bloccante per il recettore ETB. L’aldosterone invece, ha stimolato la produzione di specie reattive dell’ossigeno dopo brevi incubazioni nei fibroblasti polmonari umani, in quelli dermici sclerodermici e nelle cellule endoteliali polmonari umane ma non nei fibroblasti dermici umani normali e in quelli dermici da donatori sani. Analoghi risultati sono stati ottenuti nei fibroblasti polmonari umani, in quelli dermici sclerodermici e nelle cellule endoteliali polmonari umane stimolandoli a lungo con endotelina-1 in presenza o assenza dell’inibitore del recettore per mineralcorticoidi.Conclusioni: Questi risultati suggeriscono che l'endotelina-1 attiva i fibroblasti e la produzione di aldosterone, il vero responsabile dello stress ossidativo. L'inibizione della produzione di specie reattive dell'ossigeno dovuta all’utilizzo dell'antagonista del recettore ETB, indica che tale recettore è implicato in questo processo. Sono state trovate differenze nella produzione di stress ossidativo dopo stimolazione con endotelina-1 tra i fibroblasti dermici ottenuti da donatori sani e quelli da pazienti sclerodermici. Inoltre, negli stessi tipi cellulari, sono state evidenziate differenze nella presenza del recettore per mineralcorticoidi.Background: Pulmonary arterial hypertension is a devastating disease with an estimated patients’ survival of 68% at 1 year, 48% at 3 years and 34% at 5 years from diagnosis. The role of endothelin-1 has been studied in different conditions including systemic sclerosis. In systemic sclerosis patients, circulating endothelin- 1 is present at high levels, suggesting its involvement in the development of fibrotic and vascular damage. Indeed endothelin-1 is a potent pro-fibrotic peptide and inhibitors of endothelin-1 receptors (ETA and ETB) may exert an anti-fibrotic effect. The balance of endothelin-1 effects on smooth muscle cells and on endothelial cells is responsible for the homeostasis of vascular tone. Another interesting effect of endothelin-1 is the induction of aldosterone in endothelial cells that eventually results in the production of reactive oxygen species. Aldosterone is a key mediator of the pulmonary vascular injury response to hypoxia: mineralocorticoid receptor activation by aldosterone is involved in the remodeling/fibrosis response to hypoxia in endothelial cells in vitro and aldosterone antagonism attenuates these events.Methods: Reverse transcriptase-PCR, western blot and cytofluorimetric analyses were performed to evaluate the expression of the two isoforms of endothelin-1 receptors and mineralocorticoid receptor. Activation of fibroblast was evaluated measuring the amount of collagen-1, TGF-β and PDGF with ELISA kits. The detection of intracellular oxidative stress formation was measured with flow cytometry using fluorescence of CM-H2DCFDA after long or short incubations with endothelin-1 and its receptors inhibitors or with aldosterone and its antagonist.Results: Both endothelin-1 receptors are present in commercially available human pulmonary fibroblasts and normal human dermal fibroblasts, human dermal fibroblasts from healthy donors and scleroderma human dermal fibroblasts. Mineralocorticoid receptor expression was detected in human pulmonary fibroblasts, scleroderma human dermal fibroblasts and human pulmonary arteryendothelial cells while in normal human dermal fibroblasts and human dermal fibroblasts from healthy donors the amount of this receptor was lower than the other cells analyzed. In human pulmonary fibroblasts, scleroderma human dermal fibroblasts and human pulmonary artery endothelial cells, endothelin-1 stimulates the production of reactive oxygen species after a long incubation. In addition, the production of oxygen radicals is inhibited by the use of ETB antagonist. Interestingly, aldosterone stimulates the production of reactive oxygen species after a short incubation in human pulmonary fibroblasts, scleroderma human dermal fibroblasts and human pulmonary artery endothelial cells, but not in normal human dermal fibroblasts and human dermal fibroblasts from healthy donors. The same results were obtained by human pulmonary fibroblasts, scleroderma human dermal fibroblasts and human pulmonary artery endothelial cells stimulated with endothelin-1 for long time with mineralocorticoid receptor inhibitor.Conclusions: These results suggest that endothelin-1 activates fibroblasts and the production of aldosterone, the true responsible for oxidative stress. The inhibition of reactive oxygen species production using the ETB antagonist indicates that the ETB receptor is implicated in this process. Moreover we found differences between fibroblasts from healthy donors and from scleroderma patients in oxidative stress production after endothelin-1 stimulation. Finally we found differences in mineralocorticoid receptor presence in the same cell types

Competenze alfabetiche degli adulti calabresi

Dottorato di ricerca in Modelli di formazione, analisi teorica e comparazione, XXII ciclo, a.a. 2008-200

Varietà del'italiano nei corsi li lingua per stranieri

Dottorato di ricerca in Modelli di formazione, analisi teorica e comparazione, XXIII ciclo, A.a. 2009-2010Università della Calabria, Dipartimento di scienze dell'educazion