WIN-induced vesiculation cooperates to the inhibition of osteosarcoma cell migration

Introduction. Animal cells release vesicles that mediate the secretion of a variety of factors in the surrounding environment affecting neighboring cells. There is increasing evidence that secreted vesicles play an important role as vehicle of intercellular communication in different biological systems and are able to influence both physiological and pathological processes. Recently, we have reported that the synthetic cannabinoid WIN55,512 is able to induce osteosarcoma MG63 cell death and negatively affect cell migration. Here, we study the effects of WIN on the induction of vesicle secretion and their possible role in WIN-dependent reduction of osteosarcoma cell migratory ability. Methods. Vesicles from MG63 cells were obtained by ultracentrifuging at 140,000g media derived from cell cultures untreated and treated for 24 h with 5 uM WIN. Purified vesicles were quantified by cytofluorimetry and by detecting acetilcholinesterase activity according to established criteria. Scratch wound healing assay was employed to monitor cell migration toward the center of a gap created in a cell monolayer. Zymographic analysis was used to evaluate metalloproteinase activities in the vesicles. Results. WIN treatment induced a significant increase (about 4-fold) in the number of vesicles released by osteosarcoma cells. Wound healing assay showed that in the presence of vesicles from WIN-treated cultures, cells only partially filled the gap with respect to those conditioned with vesicles isolated from control cells which closed the gap within about 24 h. Furthermore, zymography assay showed a reduced activity of MMP-2 and MMP-9 in the vesicles obtained from WIN-treated cells. Conclusion. Data indicate that the increase in the number of vesicles released after WIN treatment and/or their probable different composition can be responsible for the relevant inhibition of MG63 cell migration induced by the cannabinoid

Enhancement of stability in systems with metastable states

The investigation of noise-induced phenomena in far from equilibrium systems is one of the approach used to understand the behaviour of physical and biological complex systems. Metastability is a generic feature of many nonlinear systems, and the problem of the lifetime of metastable states involves fundamental aspects of nonequilibrium statistical mechanics. The enhancement of the lifetime of metastable states through the noise enhanced stability effect and the role played by the resonant activation phenomenon will be discussed in models of interdisciplinary physics: (i) Ising model (ii) Josephson junction; (iii) stochastic FitzHugh-Nagumo model; (iv) a population dynamics model, and (v) a market model with stochastic volatility

Gruppo INDACO: Report della divulgazione per i progetti ASTRI e CTA Anni 2021-2022

Il gruppo INDACO (INAF per la Divulgazione di ASTRI e CTA Observatory) si è costituito alla fine del 2020 per occuparsi della divulgazione dei due progetti, ASTRI e CTA Observatory, per conto di INAF. Dopo i primi due anni di lavoro, facciamo il punto di quanto affrontato e raggiunto finora e delle prospettive future

Cinque anni di Media Inaf: report 2015-2019

In questo report, inizialmente realizzato in occasione della presentazione dei prodotti di Terza missione per la VQR 2015-2019, sono illustrate la linea editoriale della testata Media Inaf, le principali attività redazionali e i risultati conseguiti nel quinquennio di riferimento. Per i risultati quantitativi, in particolare, sono esplicitati e descritti gli indicatori utilizzati

Delega di funzioni - Quale futuro dopo il parere motivato della Ue

Gli autori analizzano la portata del parere motivato con il quale la Commissione europea ha ritenuto che la normativa interna in tema di delega di funzioni ed esonero da responsabilità del datore di lavoro in caso di delega di funzioni non sia conforme alle direttive in materia di promovimento e miglioramento della salute e sicurezza dei lavoratori durante il lavoro

Datore di lavoro. Compiti e responsabilità alla luce del Testo Unico

Gli autori delineano il quadro generale della nuova disciplina della sicurezza sul lavoro introdotta con il D.lgs n. 81/2008, mettendo in luce le più significative modifiche introdotte in tale settore, con particolare riferimento alla delega di funzioni ed al ruolo svolto dal responsabile del servizio di prevenzione e protezione

Quando la prevenzione deve salire in cattedra. Il quadro dei provvedimenti normativi, degli obblighi e delle responsabilità nelle strutture universitarie

Gli autori forniscono un quadro generale della normativa sulla sicurezza e salute nei posti di lavoro, affrontando poi la tematica concernente la specifica disciplina dettata per le università e gli istituti di istruzione universitaria in considerazione della loro peculiarità in quanto realtà nelle quali si svolgono attività di ricerca e di didattica

Disciplina penale della sicurezza sul lavoro e responsabilità amministrativa degli enti - Secondo il D.lgs. 81/08 e il D.lgs. 231/01

Nel volume viene offerto un quadro completo delle responsabilità previste dalla legge in materia di salute e sicurezza sul lavoro ed innovate con il d. lgs. 81/2008, delineando il panorama degli obblighi gravanti su tutti coloro che sono impegnati a rispettare le disposizioni in materia e delle connesse conseguenze penali ed amministrative scaturenti dalla loro inosservanza, con particolare riferimento anche alle sanzioni amministrative a carico degli enti previste dal d. lgs. 231/2001

Different modulatory effect of the synthetic cannabinoid WIN55,212-2 on tumor cell migration.

MicroRNAs are small non-coding regulatory molecules exerting pleiotropic action in different biological processes such as proliferation, differentiation, apoptosis, migration and metastasis. Deregulation of miRNA expression has been observed in various cancers, and accumulating data suggest that miRNAs can display an oncogenic, antioncogenic or an ambiguous behavior in relationship to tumor environment. In a previous research we showed that the synthetic cannabinoid WIN55,212-2 is able to reduce the migratory activity of osteosarcoma MG63 cells analyzed by means of wound healing assay. So we undertook a study to evaluate the biochemical mechanism through which WIN plays this action. To this purpose we evaluated the levels of miR-29b1, a member of miR-29 family which has been shown to impact critical steps in the migratory and metastatic cascade, such as EMT, apoptosis and angiogenesis. RT-PCR experiments showed that in MG63 cells 5 M WIN increased the level of miR-29b1 of about 700-fold. This effect was accompanied by the reduction in its putative targets MMP-2, PDGF-B and N-MYC, thus indicating that the miRNA is functionally active. Moreover, cells stably overexpressing miR-29b1 did not close the wound after 48 h, mimicking the effect of WIN in untransfected control cells. Notably, ERα(+) MCF-7 and triple negative MDA-MB-231 cells, two different breast cancer models, treated with the cannabinoid migrated into the scratched area significantly faster than the respective control cells. In these cells WIN also increased the level of miR-29b1 targets. Therefore, differently from osteosarcoma cells, these preliminary observations seem to indicate that WIN promotes migration ability in breast cancer cells. The reasons for this diverse behaviour could rely on miR-29b1, whose expression can change in different cell types or show temporal differences dictated by cell physiology and tumor microenvironment impact. Studies are in progress to shed light on the molecular mechanisms underlying this different response