FUNGAL SELF-ASSEMBLING PROTEIN LAYERS: NEW BIOTECH-TOOLS FOR BIO/NON-BIO HYBRID DEVICES

During recent years the demand of valuable scaffolds to biofunctionalize nanomaterials has been rapidly increasing. Self-assembling proteins, such as amyloid fibrils, are promising candidates for the functionalization of nanomaterials due to their chemical and mechanical stability. These fibrillar structures are typically associated to neurodegenerative diseases. However, it has been recently observed that many organisms, such as fungi and some bacteria, take advantage of the ability of polypeptides to form amyloids. Hydrophobins represent an example of functional amyloid fibrils produced at different growth stages by filamentous fungi. They are a large family of very active surface proteins and can be grouped into two distinct classes based on the stability of the amphipathic layers that they form. In particular, layers formed by class I share many structural properties with amyloid fibrils, solubilized only after harsh acid treatments Immobilization of active proteins on the hydrophobin layer has been proven as a simple approach for the functionalization of different surfaces useful in biotechnological applications. For most of these applications, a critical aspect concerns protein orientation with respect to the surface. To overcome this problem, design and recombinant production of fused engineered proteins combining the adhesive moiety of hydrophobins to target proteins can be a valuable choice. This PhD project has been focused on recombinant production in different hosts (Pichia pastoris and Escherichia coli) of chimera proteins built by selected target proteins (laccase, antimicrobial peptides and antibody) fused to an adhesive self-assembling class I hydrophobin, and on their exploitation in some application fields. The selected class I hydrophobin was Vmh2, extracted from the mycelium of the fungus Pleurotus ostreatus, one of the most hydrophobic hydrophobin able to spontaneously form stable and homogeneous layers on hydrophilic or hydrophobic surfaces, changing their wettability. As a first goal, the hydrophobin Vmh2 was fused to the laccase POXA1b from P. ostreatus, a high redox potential oxidase endowed with a remarkable stability at high temperature and at alkaline pH. The resulting fusion enzyme was secreted into the culture medium by the heterologous yeast P. pastoris and directly used for coating different surfaces, i.e. graphene and polystyrene, without additional purification steps. The immobilized enzyme was exploited to develop innovative optical and electrochemical biosensing platform for the detection of phenolic compounds. Subsequently, two chimeric proteins were recombinantly expressed in E. coli: in the first case, Vmh2 was fused to the antimicrobial peptide LL37, a small cationic amphiphilic peptide belonging to the family of cathelicidins which has an effective activity against a wide range of bacteria and fungi. The LL37-Vmh2 chimeric protein was successfully produced and its adhesive and antibiofilm capabilities, against Staphylococcus epidermidis, tested by coating polystyrene surfaces. The chimera HN1-Vmh2, the fusion between the hydrophobin and the anti-mesothelin ScFv antibody, was produced in the soluble fraction of E. coli whilst its purification and exploiation needs to be optimized. In conclusion, chimeric proteins obtained by the genetic fusion of the hydrophobin Vmh2 to target proteins represent a proof of concept of versatile and straightforward solutions of surface functionalizations that could be explored in several fields. Moreover, during the six months stay at the CNRS of Grenoble, the work was focused on carbon nanotubes (CNT) functionalization using the native laccase POXC from P. ostreatus. POXC is a promising biocathode for enzymatic biofuel cells (EBFCs), since it shows efficient ORR (Oxygen Reduction Reaction) performances and it is capable to operate in a conventional H2/air proton-exchange membrane fuel cell

Guida per gestire il tempo in maniera efficace

Indice: Sezione 1: Gestire il tempo. La allocazione dei tempi. Preparare un piano. Attivita'  1: Allocare i tempi. I picchi personali di performance. Attivita' 2: Il vostro momento di picco. Definizione degli obiettivi. Attivita' 3: Controllare i tempi. Identificare le priorita' . Attivita'  4: Programmare il tempo. La delega come strumento per un uso piu' efficace del tempo e delle risorse disponibili. Attivita' 5: Delegare per la massimizzazione della risorsa tempo. Attivita' 6: La delega (role-playing) Il controllo del tempo. Attivita' 7: Ridurre al minimo le interruzioni. Gli sprechi di tempo. Sezione 2: Lavorare sotto pressione, evitando lo stress. Quando lo stress diventa ansia. I sintomi dello stress. Attivita' 1: I sintomi dello stress. Lo stress come questione organizzativa. Attivita'   2: Lo stress e il vostro lavoro. Come ridurre le cause di stress. Assertivita'. Attivita' 3: Stress e assertivita'. Lavorare in grupp

Again-st Abandon. National Report Index

Lo studio e' stato realizzato da Stoa' e Studio Staff nell'ambito del progetto "AGAIN-ST ABANDON Contrastare il fenomeno degli abbandoni, ripensare i linguaggi della consulenza". Il progetto Again-st Abandon, finanziato dal Programma europeo "Leonardo da Vinci", si pone l'obiettivo di contrastare il fenomeno dei drop-out (DO) - quei giovani che interrompono il percorso scolastico e formativo rendendosi invisibili agli operatori del sociale - nell'ottica di un loro reinserimento nei contesti formali di istruzione, formazione ed impiego. Nel paper viene sviluppata un'analisi territoriale dell'area del Comune di Ercolano tesa ad indagare la domanda e l'offerta di servizi di orientamento e formazione, oltre alla visibilita' e all'accessibilita' degli stessi, attraverso l'utilizzo di dati statistici e indicatori di tipo qualitativo

Le azioni a favore della crescita del potenziale di innovazione all'interno di sistemi territoriali

Indice: Inquadramento teorico delle politiche a favore dei sistemi territoriali. Casi di studio: Ristrutturazione industriale e sviluppo locale il caso di Sesto San Giovanni; Il modello Stoa' Progetto pilota 4 circoscrizion

Transfer of H2O2 from Mitochondria to the endoplasmic reticulum via Aquaporin-11

Some aquaporins (AQPs) can transport H2O2 across membranes, allowing redox signals to proceed in and between cells. Unlike other peroxiporins, human AQP11 is an endoplasmic reticulum (ER)-resident that can conduit H2O2 to the cytosol. Here, we show that silencing Ero1α, an ER flavoenzyme that generates abundant H2O2 during oxidative folding, causes a paradoxical increase in luminal H2O2 levels. The simultaneous AQP11 downregulation prevents this increase, implying that H2O2 reaches the ER from an external source(s). Pharmacological inhibition of the electron transport chain reveals that Ero1α downregulation activates superoxide production by complex III. In the intermembrane space, superoxide dismutase 1 generates H2O2 that enters the ER channeled by AQP11. Meanwhile, the number of ER-mitochondria contact sites increases as well, irrespective of AQP11 expression. Taken together, our findings identify a novel interorganellar redox response that is activated upon Ero1α downregulation and transfers H2O2 from mitochondria to the ER via AQP11.This work was supported through grants from the Associazione Italiana Ricerca sul Cancro (IG 2019-23285 to R.S.) and Ministero dell Istruzione, dell Università e della Ricerca (MIUR)-PRIN (grant no. 2017XA5J5N). I.M.-F. was supported by the Madrid Government (Comunidad de Madrid) under the Multiannual Agreement with UC3M in the line of Research Funds for Beatriz Galindo Fellowships (REDOXSKIN-CM-UC3M), and in the context of the V PRICIT (Regional Programme of Research and Technological Innovation", and by Proyectos de I + D + I (PID2020-114230 GA-I00 to I.M.-F.) funded by MCIN/AEI/10.13039/501100011033/. MG was supported by the European Unions Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 754432 and the Polish Ministry of Science and Higher Education, from financial resources for science in 2018 2023 granted for the implementation of an international co-financed project

Ricerca preliminare sui fabbisogni formativi per la definizione della figura professionale del Manager dello sviluppo locale

Indice della ricerca: Introduzione. L'esperienza del Master in International Development: l'azione locale nella prospettiva internazionale. L'evoluzione dell'esperienza. Il Master Operatori allo Sviluppo Gli obiettivi della ricerca. L'analisi di campo Definizione ed interpretazione della figura professionale. Premessa. La dimensione locale dei processi di sviluppo. L'agente di sviluppo: un profilo strattonato. Verso una comunita' professionale? I percorsi formativi della figura professionale. Premessa. La ricognizione dell'offerta formatica post laurea in materia di sviluppo locale. La metodologia. La lettura incrociata dei dati e la comparazione con il Master in Local Development. I percorsi di carriera dei diplomati Master. Premessa. Alcuni casi di successo L'analisi di campo sulla figura professionale del Manager dello sviluppo L'approccio metodologico. L'analisi dei dati. Una lettura dei dati per tipologie di intervistati. La figura professionale del Manager dello sviluppo locale: un tentativo di sintesi. Bibliografi

Divergent Effect of Cobalt and Beryllium Salts on the Fate of Peripheral Blood Monocytes and T Lymphocytes

Occupational exposure to metals such as cobalt and beryllium represents a risk factor for respiratory health and can cause immune-mediated diseases. However, the way they act may be different. We show here that the two metals have a divergent effect on peripheral T lymphocytes and monocytes: BeSO4 induces cell death in monocytes but not in T lymphocytes, which instead respond by producing Interferon gamma (IFN-γ); conversely, CoCl2 induces apoptosis in T lymphocytes but not in monocytes. Interestingly, both metals induce p53 overexpression but with a dramatic different outcome. This is because the effect of p53 in CoCl2-treated monocytes is counteracted by the antiapoptotic activity of cytoplasmic p21Cip1/WAF1, the activation of nuclear factor κB, and the inflammasome danger signaling pathway leading to the production of proinflammatory cytokines. However, CoCl2-treated monocytes do not fully differentiate into macrophage or dendritic cells, as inferred by the lack of expression of CD16 and CD83, respectively. Furthermore, the expression of HLA-class II molecules, as well as the capability of capturing and presenting the antigens, decreased with time. In conclusion, cobalt keeps monocytes in a partially activated, proinflammatory state that can contribute to some of the pathologies associated with the exposure to this meta

Human aquaporin-11 guarantees efficient transport of H2O2 across the endoplasmic reticulum membrane

Hydrogen peroxide (H2O2) is an essential second intracellular messenger. To reach its targets in the cytosol, H2O2 must cross a membrane, a feat that requires aquaporins (AQP) endowed with 'peroxiporin' activity (AQP3, AQP8, AQP9). Here, we exploit different organelle-targeted H2O2-sensitive probes to show that also AQP11 efficiently conduits H2O2. Unlike other peroxiporins, AQP11 is localized in the endoplasmic reticulum (ER), accumulating partly in mitochondrial-associated ER membranes (MAM). Its downregulation severely perturbs the flux of H2O2 through the ER, but not through the mitochondrial or plasma membranes. These properties make AQP11 a potential regulator of ER redox homeostasis and signaling.This work was supported in part through grants from the Associazione Italiana Ricerca sul Cancro (IG 2016-18824 to R.S.), the Fondazione Cariplo (2015-0591 to R.S.), and the Telethon (GGP15059 to R.S.). A.R. was supported by local funds from the University of Ferrara and the Italian Ministry of Health (GR-2016-02364602). Italian Association for Cancer Research (AIRC, IG-18624), Telethon (GGP11139B), and local funds from the University of Ferrara to P.P

A Health Technology Assessment in Maxillofacial Cancer Surgery by Using the Six Sigma Methodology

Squamous cell carcinoma represents the most common cancer affecting the oral cavity. At the University of Naples “Federico II”, two different antibiotic protocols were used in patients undergoing oral mucosa cancer surgery from 2006 to 2018. From 2011, there was a shift; the combination of Cefazolin plus Clindamycin as a postoperative prophylactic protocol was chosen. In this paper, a health technology assessment (HTA) is performed by using the Six Sigma and DMAIC (Define, Measure, Analyse, Improve, Control) cycle in order to compare the performance of the antibiotic protocols according to the length of hospital stay (LOS). The data (13 variables) of two groups were collected and analysed; overall, 136 patients were involved. The American Society of Anaesthesiologist score, use of lymphadenectomy or tracheotomy and the presence of infections influenced LOS significantly (p-value < 0.05) in both groups. Then, the groups were compared: the overall difference between LOS of the groups was not statistically significant, but some insights were provided by comparing the LOS of the groups according to each variable. In conclusion, in light of the insights provided by this study regarding the comparison of two antibiotic protocols, the utilization of DMAIC cycle and Six Sigma tools to perform HTA studies could be considered in future research

Risorsa cultura: modelli internazionali di management dei beni culturali

Indice: Beni culturali e sviluppo del territorio Il ruolo della cultura nella provincia di Napoli Stoa' per il management dei beni culturali RSO ed i beni culturali Le best practice L'idea progettuale. Considerazioni preliminari. Lo study tour come strumento di formazione Partecipanti al progetto Esperti che hanno partecipato al progetto I progetti: La valorizzazione culturale del waterfront dell'area di Napoli est. Dal vetro al cristallo. Dal decentramento al coordinamento: le biblioteche comunali di Napoli. Pianificazione e controllo dell'offerta culturale. La rete delle realta' culturali... in Comune. Ecomuseo del monte Somma: l'altra faccia del Vesuvio. Siti e miti delle sirene. Santi in provincia. Rete di qualita' integrata dei servizi culturali dei comuni a nord di Napoli. Beni culturali e vantaggio competitivo territorial