Axitinib induces DNA damage response leading to senescence, mitotic catastrophe, and increased NK cell recognition in human renal carcinoma cells.

Tyrosine kinase inhibitors (TKIs) including axitinib have been introduced in the treatment of renal cell carcinoma (RCC) because of their anti-angiogenic properties. However, no evidence are presently available on a direct cytotoxic anti-tumor activity of axitinib in RCC.Herein we reported by western blot analysis that axitinib treatment induces a DNA damage response (DDR) initially characterized by γ-H2AX phosphorylation and Chk1 kinase activation and at later time points by p21 overexpression in A-498 and Caki-2 RCC cells although with a different potency. Analysis by immunocytochemistry for the presence of 8-oxo-7,8-dihydro-2'-deoxyguanosine in cellular DNA and flow cytometry using the redox-sensitive fluorescent dye DCFDA, demonstrated that DDR response is accompanied by the presence of oxidative DNA damage and reactive oxygen species (ROS) generation. This response leads to G2/M cell cycle arrest and induces a senescent-like phenotype accompanied by enlargement of cells and increased senescence-associated β-galactosidase activity, which are abrogated by N-acetyl cysteine (NAC) pre-treatment. In addition, axitinib-treated cells undergo to cell death through mitotic catastrophe characterized by micronucleation and abnormal microtubule assembly as assessed by fluorescence microscopy.On the other hand, axitinib, through the DDR induction, is also able to increase the surface NKG2D ligand expression. Accordingly, drug treatment promotes NK cell recognition and degranulation in A-498 RCC cells in a ROS-dependent manner.Collectively, our results indicate that both cytotoxic and immunomodulatory effects on RCC cells can contribute to axitinib anti-tumor activity

Capsaicin triggers autophagic cell survival which drives epithelial mesenchymal transition and chemoresistance in bladder cancer cells in an Hedgehog-dependent manner

Bladder cancer (BC) is a common urologic tumor characterized by high risk of recurrence and mortality. Capsaicin (CPS), used as an intravesical drug for overactive bladder, was demonstrated to induce cell death in different cancer cells including BC cells.Here we found that treatment of high-grade BC cells with high dose of CPS triggers autophagy. Infact, the CPS treatment alters the redox homeostasis by inducing production of radicals, mitochondrial depolarization, alterations of ADP/ATP ratio and activation of AMPK pathway stimulating the autophagic process in BC cells. The inhibition of autophagy, by using the specific inhibitor bafilomycin A or Beclin 1 knock-down, enhanced the CPS-induced cell death, demonstrating that CPS-induced autophagy acts as a pro-survival process in BC cells. By using PCR arrays and FACS analysis, we found that the CPS-treated BC cells displayed typical mesenchymal features of the epithelial mesenchymal transition (EMT) as elongated shape and over-expression of vimentin, α5 and β1 integrin subunits, integrin-like kinase and the anti-apoptotic Bcl-2 proteins. Moreover, we demonstrated that CPS treatment stimulates upregulation of Dhh/Ptch2/Zeb2 members of the Hedgehog signaling pathway, increases CD24, VEGFA and TIMP1 and decreases CD44 and ALCAM mRNA expression levels. By PTCH2 knock-down we found that the Hedgehog signaling pathway is involved in the CPS-induced autophagy and EMT phenotype.Finally, we also showed that the CPS-resistant EMT-positive BC cells displayed an increased drug-resistance to the cytotoxic effects of mitomycin C, gemcitabine and doxorubicine drugs commonly used in BC therapy

Axitinib induces senescence-associated cell death and necrosis in glioma cell lines: The proteasome inhibitor, bortezomib, potentiates axitinib-induced cytotoxicity in a p21(Waf/Cip1) dependent manner.

Glioblastoma is associated with a poor overall survival despite new treatment advances. Antiangiogenic strategies targeting VEGF based on tyrosine kinase inhibitors (TKIs) are currently undergoing extensive research for the treatment of glioma. Herein we demonstrated that the TKI axitinib induces DNA damage response (DDR) characterized by γ-H2AX phosphorylation and Chk1 kinase activation leading to G2/M cell cycle arrest and mitotic catastrophe in U87, T98 and U251 glioma cell lines. Moreover, we found that p21(Waf1/Cip1) increased levels correlates with induction of ROS and senescence-associated cell death in U87 and T98 cell lines, which are reverted by N-acetyl cysteine pretreatment. Conversely, U251 cell line showed a resistant phenotype in response to axitinib treatment, as evidenced by cell cycle arrest but no sign of cell death. The combinatorial use of axitinib with other therapies, with the aim of inhibiting multiple signaling pathways involved in tumor growth, can increase the efficiency of this TKI. Thus, we addressed the combined effects of axitinib with no toxic doses of the proteasome inhibitor bortezomib on the growth of U87 and T98 axitinib- sensitive and axitinib-resistant U251 cell lines. Compared to single treatments, combined exposure was more effective in inhibiting cell viability of all glioma cell lines, although with different cell death modalities. The regulation of key DDR and cell cycle proteins, including Chk1, γ-H2AX and p21(Waf1/Cip1) was also studied in glioma cell lines. Collectively, these findings provide new perspectives for the use of axitinib in combination with Bortezomib to overcome the therapy resistance in gliomas

MicroRNA-222 regulates muscle alternative splicing through Rbm24 during differentiation of skeletal muscle cells

A number of microRNAs have been shown to regulate skeletal muscle development and differentiation. MicroRNA-222 is downregulated during myogenic differentiation and its overexpression leads to alteration of muscle differentiation process and specialized structures. By using RNA-induced silencing complex (RISC) pulldown followed by RNA sequencing, combined with in silico microRNA target prediction, we have identified two new targets of microRNA-222 involved in the regulation of myogenic differentiation, Ahnak and Rbm24. Specifically, the RNA-binding protein Rbm24 is a major regulator of muscle-specific alternative splicing and its downregulation by microRNA-222 results in defective exon inclusion impairing the production of muscle-specific isoforms of Coro6, Fxr1 and NACA transcripts. Reconstitution of normal levels of Rbm24 in cells overexpressing microRNA-222 rescues muscle-specific splicing. In conclusion, we have identified a new function of microRNA-222 leading to alteration of myogenic differentiation at the level of alternative splicing, and we provide evidence that this effect is mediated by Rbm24 protei

Remote landslide mapping using a laser rangefinder binocular and GPS

We tested a high-quality laser rangefinder binocular coupled with a GPS receiver connected to a Tablet PC running dedicated software to help recognize and map in the field recent rainfall-induced landslides. The system was tested in the period between March and April 2010, in the Monte Castello di Vibio area, Umbria, Central Italy. To test the equipment, we measured thirteen slope failures that were mapped previously during a visual reconnaissance field campaign conducted in February and March 2010. For reference, four slope failures were also mapped by walking the GPS receiver along the landslide perimeter. Comparison of the different mappings revealed that the geographical information obtained remotely for each landslide by the rangefinder binocular and GPS was comparable to the information obtained by walking the GPS around the landslide perimeter, and was superior to the information obtained through the visual reconnaissance mapping. Although our tests were not exhaustive, we maintain that the system is effective to map recent rainfall induced landslides in the field, and we foresee the possibility of using the same (or similar) system to map landslides, and other geomorphological features, in other areas

Identification and mapping of recent rainfall-induced landslides using elevation data collected by airborne Lidar

International audienceA high resolution Digital Elevation Model with a ground resolution of 2 m×2 m (DEM2) was obtained for the Collazzone area, central Umbria, through weighted linear interpolation of elevation points acquired by Airborne Lidar Swath Mapping. Acquisition of the elevation data was performed on 3 May 2004, following a rainfall period that resulted in numerous landslides. A reconnaissance field survey conducted immediately after the rainfall period allowed mapping 70 landslides in the study area, for a total landslide area of 2.7×105 m2. Topographic derivative maps obtained from the DEM2 were used to update the reconnaissance landslide inventory map in 22 selected sub-areas. The revised inventory map shows 27% more landslides and 39% less total landslide area, corresponding to a smaller average landslide size. Discrepancies between the reconnaissance and the revised inventory maps were attributed to mapping errors and imprecision chiefly in the reconnaissance field inventory. Landslides identified exploiting the Lidar elevation data matched the local topography more accurately than the same landslides mapped using the existing topographic maps. Reasons for the difference include an incomplete or inaccurate view of the landslides in the field, an unfaithful representation of topography in the based maps, and the limited time available to map the landslides in the field. The high resolution DEM2 was compared to a coarser resolution (10 m×10 m) DEM10 to establish how well the two DEMs captured the topographic signature of landslides. Results indicate that the improved topographic information provided by DEM2 was significant in identifying recent rainfall-induced landslides, and was less significant in improving the representation of stable slopes

Dell'attenzione. Fenomenologia ed etica per la relazione medico-paziente in ambiente digitale.

La relazione medico-paziente sta vivendo un momento di grande difficoltà: aumentano le opportunità diagnostiche e terapeutiche, ci sono maggiori strumentazioni tecniche, ma al tempo stesso diminuiscono le occasioni di incontro, si impoverisce la comunicazione e sempre più pazienti lamentano di non sentirsi pienamente accolti nel processo di cura. Le difficoltà che caratterizzano la relazione di cura hanno una storia lunga, e sembrano essersi esacerbate nel corso del Novecento, ma di certo oggi sono esposte a sfide inedite di fronte al pervasivo impatto delle nuove tecnologie sanitarie. Lo scenario contemporaneo presenta, in particolar modo, due principali rischi: da un lato, si crea uno sbilanciamento tra le possibilità del to cure e le occasioni del to care, ovvero tra l’efficientamento delle prestazioni tecniche impiegate per la cura del corpo malato e l’opportunità di farsi carico delle ben più complesse dimensioni esistenziali che connotano l’accoglienza e la cura della persona; dall’altra, da parte dei medici, si incentivano dei riduzionismi nel modo di “percepire” la persona malata e trattare la sua storia di malattia, oppure si ripropongono nuove riduzioni quando la cura viene riallocata negli ambienti digitali della salute. Nel crinale di tali questioni nasce l’appello della persona malata a non essere ridotta alle occorrenze del corpo o della propria malattia, ma anzi ad essere considerata con attenzione. Ciò sollecita a rileggere le condizioni di possibilità della relazione, a ripensare certe prassi relazionali e risemantizzare alcuni concetti propri del lessico medico. Lo scopo di questa ricerca è quello di analizzare, secondo un approccio fenomenologico ed ermeneutico, quali siano i caratteri del rapporto medico-paziente, quali rilievi etici emergono negli ambienti digitali della salute e come questo rapporto possa essere riletto sotto la lente del concetto di attenzione. Il tema dell’attenzione, infatti, si rivela particolarmente adeguato a questo proposito per diversi motivi: anzitutto perché permette di inquadrare l’analisi dentro un orizzonte etico che non guarda semplicemente a cosa i medici debbano prestare attenzione, ma soprattutto a come farlo e secondo quali posture relazionali eticamente accettabili; in secondo luogo, perché l’attenzione rappresenta un concetto nomade ed ibrido, un grande tema mancato del pensiero morale classico e moderno, che costringe sia ad un suo rigoroso approfondimento concettuale sia ad una sua applicazione nella medicina; infine, perché proprio l’attenzione alla persona è ciò che rischia di venire impoverita quando l’intermediazione del digitale e delle nuove economie dell’attenzione la frammentano o esauriscono negli ambienti digitali della salute. Per questi motivi, all’interno di un quadro articolato e in costante mutamento, l’indagine si strutturata seguendo tre principali scansioni, tese a delineare la fenomenologia della persona malata, il concetto di attenzione e degli ambienti digitali nei quali si presta attenzione alla persona. A partire da una ricostruzione fenomenologica dell’esistenza che ha visto il coinvolgimento di differenti fenomenologi quali Husserl, Merleau-Ponty, Kay Toombs, Carel, Costa e Cesana, la ricerca ha posto una distinzione preliminare tra malattia (disease) ed esperienza di malattia (illness), tra la considerazione organica o bio-fisiologica del corpo materiale (Körper) e la dimensione percettiva, esistenziale e incarnata dalla coscienza del corpo vissuto (Leib). Anche sotto il profilo epistemologico, si è indagato come intendere i concetti di salute e malattia (Canguilhem, Gadamer) al fine di distinguere, senza separare, la prospettiva promossa dallo sguardo medico con l’istanza di salute proveniente dal soggetto vulnerabile. Non riducendola alla deviazione da una norma statistica, né al mancato assolvimento di una funzione fisiologica o di un meccanismo biochimico del corpo, l’esperienza di malattia rivela una dimensione carica di significati. E altrettanto il corpo, mai ridotto a un sostrato materiale come nel corpo morto, svela la dimensione esistenziale del corpo vivo: il luogo privilegiato in cui “prende corpo” l’esperienza di malattia. Saper prestare attenzione a tali dimensioni non è un atteggiamento spontaneo per i professionisti della salute, soprattutto a seguito di un percorso formativo che li ha visti riconfigurare la percezione e l’epistemologia del proprio sguardo medico (Foucault, Good); eppure tali considerazioni permettono di ripensare la postura clinica della medicina e della diagnosi. Dopo aver ricostruito e analizzato la fenomenologia dell’attenzione, che la vede strettamente legata alla percezione e al mondo della coscienza, la ricerca si è proposta di risemantizzare il significato stesso di attenzione, non riducendolo al significato spesso incentivato dalle scienze cognitive e inteso perlopiù come un’opera di selezione percettiva, focalizzazione o concentrazione. Si è voluto offrire una semantica ampia del concetto sulla scorta della filosofia morale e dell’etica della cura, come sinonimo di considerazione personale, riconoscimento, interesse e partecipazione coinvolta a ciò verso cui si presta attenzione. Ciò ha permesso di delineare pure una diversa epistemologia dell’attenzione nella clinica che sembra sempre abitare sulla soglia degli eventi tra passività e attività, tra il pathos dell’attenzione che viene a noi e la risposta di considerazione che viene da noi (Waldenfels). Si è così proceduto ad approfondire gli scenari odierni della medicina al fine di approcciare direttamente la questione inedita degli ambienti digitali della salute, nei quali nascono benefici ed effetti collaterali. In tali ambienti non solo si riscrivono occasioni, ma si delineano nuove condizioni di (im)possibilità: il dettato dell’incontro clinico è ripensato, i protagonisti della cura sono riallocati in nuovi ambienti, la figura e il sapere medico sembrano essere al tempo stesso ovunque e in nessun luogo. In particolare, con il costante processo di datificazione informazionale, si assiste a un’epocale riscrittura dei concetti di corpo vivo, di salute e di persona malata, dei quali la ricerca ne ha mostrato le ripercussioni etiche, le conseguenze sul percorso di cura e i paradossi che possono crearsi sul piano dell’esperienza. Perciò si è analizzato come il digitale modifica i modi dell’attenzione alla persona, ad esempio quando un teleconsulto medico è intermediato da uno schermo digitale o quando la mole informazionale supera e disperde le capacità attentive del medico. Questi scenari spingono a valutare la digitalizzazione della medicina come un processo di efficientamento, ma non privo di rischi; un’opportunità da saper concertare senza lasciarsi assuefare dalle false promesse di un futuro utopico. L’attenzione dimostra una rilevanza etica anche quando diventa sinonimo di cautela e pensiero critico nei confronti dei cambiamenti in atto, dell’umano e di tutto ciò che si reputa ancora importante per l’esistenza, singolare e collettiva

Bio-based building components: A newly sustainable solution for traditional walls made of Arundo donax and gypsum

To contribute to the use of bio-based materials in the building sector, a novel bio-based wall panel, with a high thermal performance level, is proposed in this work. The panel is based on an ancient rural technique, widely diffused in southern Italy, which makes use of Arundo donax L. canes combined with gypsum plaster to build walls and ceilings of rural buildings. The enhancement of the thermal capacity of these panels by means of the introduction in the canes of a natural wax oleogel (WO) is proposed in this paper. A specific experimental campaign aiming at the comparison of traditional and innovative panels was carried out to assess the enhanced thermal performance of the proposed solution. The maximum value of heat flow absorbed from the panel with WO was 61.08 W/m(2) around a mean panel temperature of 24 & DEG;C, corresponding to the melting temperature range of the WO. The panel without WO at the same temperature absorbed an incoming heat flow of 34.64 W/m(2) which is about 57% of the panel with WO. The panel with WO released at a temperature of about 27.5 & DEG;C, a heat flow of 43.42 W/m(2). At the same temperature of about 27.5 & DEG;C, the panel without WO released a heat flow of 34.38 W/m(2) which is about 80% that of the panel with WO. The results highlighted that the addition of natural WO has enhanced the thermal capacity of the panel facilitating heat dissipation through the borders. These characteristics make the panel a suitable component for internal partitions of controlled temperature zones such as residential rooms, storage food areas, livestock buildings, and where it is necessary to obtain a constant environmental temperature. In particular, the null or low toxicity of the panel's materials allows for partition use, also in hygienically safe environments