Proteomic Analysis Reveals a Mitochondrial Remodeling of βTC3 Cells in Response to Nanotopography

Recently, using cluster-assembled zirconia substrates with tailored roughness produced by supersonic cluster beam deposition, we demonstrated that \u3b2 cells can sense nanoscale features of the substrate and can translate these stimuli into a mechanotransductive pathway capable of preserveing \u3b2-cell differentiation and function in vitro in long-term cultures of human islets. Using the same proteomic approach, we now focused on the mitochondrial fraction of \u3b2TC3 cells grown on the same zirconia substrates and characterized the morphological and proteomic modifications induced by the nanostructure. The results suggest that, in \u3b2TC3 cells, mitochondria are perturbed by the nanotopography and activate a program involving metabolism modification and modulation of their interplay with other organelles. Data were confirmed in INS1E, a different \u3b2-cell model. The change induced by the nanostructure can be pro-survival and prime mitochondria for a metabolic switch to match the new cell needs

MosChito rafts as effective and eco-friendly tool for the delivery of a Bacillus thuringiensis-based insecticide to Aedes albopictus larvae

Adult mosquito females, through their bites, are responsible for the transmission of different zoonotic pathogens. Although adult control represents a pillar for the prevention of disease spread, larval control is also crucial. Herein we characterized the effectiveness of a suitable tool, named "MosChito raft", for the aquatic delivery of a Bacillus thuringiensis var. israelensis (Bti) formulate, a bioinsecticide active by ingestion against mosquito larvae. MosChito raft is a floating tool composed by chitosan cross-linked with genipin in which a Bti-based formulate and an attractant have been included. MosChito rafts (i) resulted attractive for the larvae of the Asian tiger mosquito Aedes albopictus, (ii) induced larval mortality within a few hours of exposure and, more importantly, (iii) protected the Bti-based formulate, whose insecticidal activity was maintained for more than one month in comparison to the few days residual activity of the commercial product. The delivery method was effective in both laboratory and semi-field conditions, demonstrating that MosChito rafts may represent an original, eco-based and user-friendly solution for larval control in domestic and peri-domestic aquatic habitats such as saucers and artificial containers in residential or urban environments

Proteomic dissection of nanotopography-sensitive mechanotransductive signaling hubs that foster neuronal differentiation in PC12 cells

Neuronal cells are competent in precisely sensing nanotopographical features of their microenvironment. The perceived microenvironmental information will be \ue2\u80\u9cinterpreted\ue2\u80\u9d by mechanotransductive processes and impacts on neuronal functioning and differentiation. Attempts to influence neuronal differentiation by engineering substrates that mimic appropriate extracellular matrix (ECM) topographies are hampered by the fact that profound details of mechanosensing/-transduction complexity remain elusive. Introducing omics methods into these biomaterial approaches has the potential to provide a deeper insight into the molecular processes and signaling cascades underlying mechanosensing/-transduction but their exigence in cellular material is often opposed by technical limitations of major substrate top-down fabrication methods. Supersonic cluster beam deposition (SCBD) allows instead the bottom-up fabrication of nanostructured substrates over large areas characterized by a quantitatively controllable ECM-like nanoroughness that has been recently shown to foster neuron differentiation and maturation. Exploiting this capacity of SCBD, we challenged mechanosensing/-transduction and differentiative behavior of neuron-like PC12 cells with diverse nanotopographies and/or changes of their biomechanical status, and analyzed their phosphoproteomic profiles in these settings. Versatile proteins that can be associated to significant processes along the mechanotransductive signal sequence, i.e., cell/cell interaction, glycocalyx and ECM, membrane/f-actin linkage and integrin activation, cell/substrate interaction, integrin adhesion complex, actomyosin organization/cellular mechanics, nuclear organization, and transcriptional regulation, were affected. The phosphoproteomic data suggested furthermore an involvement of ILK, mTOR, Wnt, and calciumsignaling in these nanotopography- and/or cell mechanics-related processes. Altogether, potential nanotopography-sensitive mechanotransductive signaling hubs participating in neuronal differentiation were dissected

Scale invariant disordered nanotopography promotes hippocampal neuron development and maturation with involvement of mechanotransductive pathways

The identification of biomaterials which promote neuronal maturation up to the generation of integrated neural circuits is fundamental for modern neuroscience. The development of neural circuits arises from complex maturative processes regulated by poorly understood signaling events, often guided by the extracellular matrix (ECM). Here we report that nanostructured zirconia surfaces, produced by supersonic cluster beam deposition of zirconia nanoparticles and characterized by ECM-like nanotopographical features, can direct the maturation of neural networks. Hippocampal neurons cultured on such cluster-assembled surfaces displayed enhanced differentiation paralleled by functional changes. The latter was demonstrated by single-cell electrophysiology showing earlier action potential generation and increased spontaneous postsynaptic currents compared to the neurons grown on the featureless unnaturally flat standard control surfaces. Label-free shotgun proteomics broadly confirmed the functional changes and suggests furthermore a vast impact of the neuron/nanotopography interaction on mechanotransductive machinery components, known to control physiological in vivo ECM-regulated axon guidance and synaptic plasticity. Our results indicate a potential of cluster-assembled zirconia nanotopography exploitable for the creation of efficient neural tissue interfaces and cell culture devices promoting neurogenic events, but also for unveiling mechanotransductive aspects of neuronal development and maturation

Scene di fine Ottocento. L'Italia fin de si\ue8cle a teatro

Muovendosi fra palcoscenici e platee , fra editori e imprenditori teatrali, fra interpreti, traduttori e autori di generi vecchi e nuovi il volume mostra un mondo teatrale multiforme e complesso che nei decenni a cavallo tra 800 e 900 sta adeguandosi ai cambiamenti della modernit\ue0, a una pi\uf9 diffusa domanda di intrattenimento e all'esigenza di definire gerarchie culturali inedite

Neuronal Cells Confinement by Micropatterned Cluster-Assembled Dots with Mechanotransductive Nanotopography

Artificially grown neuronal cultures of brain cells have been used for decades in the attempt to reproduce and study in vitro the complexity of brain circuits. It soon became evident that this alone was insufficient, because of the random architecture of these artificial networks. Important groundwork therefore resulted in the development of methods to confine neuronal adhesion at specific locations to match predefined network topologies and connectivity. Despite this notable progress in neural circuitry engineering, there is still need for micropatterned substrates that recapitulate better biophysical cues of the neuronal microenvironment, taking into account recent findings of their significance for neuronal differentiation and functioning. Here, we report the development and characterization of a novel approach that, by using supersonic cluster beam deposition of zirconia nanoparticles, allows the patterning of small nanostructured cell-adhesive areas according to predefined geometries onto elsewhere nonadhesive antifouling glass surfaces. As distinguishing features, compared to other micropatterning approaches in this context, the integrated nanostructured surfaces possess extracellular matrix-like nanotopographies of predetermined roughness; previously shown to be able to promote neuronal differentiation due to their impact on mechanotransductive processes, and can be used in their original state without any coating requirements. These micropatterned substrates were validated using (i) a neuron-like PC12 cell line and (ii) primary cultures of rat hippocampal neurons. After initial uniform plating, both neuronal cells types were found to converge and adhere specifically to the nanostructured regions. The cell-adhesive areas effectively confined cells, even when these were highly mobile and repeatedly attempted to cross boundaries. Inside these small permissive islands, cells grew and differentiated, in the case of the hippocampal neurons, up to the formation of mature, functionally active, and highly connected synaptic networks. In addition, when spontaneous instances of axon bridging between nearby dots occurred, a functional interdot communication between these subgroups of cells was observed

Nanostructured zirconia surface induce differentiation and maturation events in neonatal neuronal cells from rat hippocampus

BACKGROUND-AIM Biomaterials with a positive impact on neurogenic processes could give a valuable contribution to prospective cell replacement therapies for neurodegenerative diseases. The idea behind is the in vitro recapitulation of neurogenesis for the substitution of damaged neurons by healthy ones. In general, the capacity of biomaterials to favour differentiation-promoting effects relies on the cellular competence for mechanotransduction, the perception of biophysical signals (i.e. rigidity and topography) and their conversion into biochemical cellular responses. So far most approaches in the context of neuronal cells were based on the modulation of substrate rigidity. Here we present the possibility to promote neuron differentiation and maturation by appropriate nanorough zirconia surfaces produced by supersonic cluster beam deposition (SCBD). METHODS Via quantitative shotgun proteomic analysis, we compared the protein profile of hippocampal cells interacting for 3 days with the differentiation-promoting nanorough zirconia surface with the cells grown on glass kept as control. An Anova test (FDR 0.05) was carried out to identify proteins differently expressed among different conditions. RESULTS In total almost 600 proteins were altered in their expression level. There was a quite drastic change of the cellular program versus mature neurons. Infact, neonatal rat hippocampal neuronal cells showed accelerated and enhanced maturation and synaptogenesis on these surfaces. This was demonstrated by the earlier and stronger presence of synaptic markers, electrophysiological activity on the single cell and network level and a protein profile confirming an advanced state of neurogenic events. Congruent to the nature of the biophysical signal input, the proteomic data also suggested a strong involvement of cytoskeleton- and integrin adhesome-related processes. CONCLUSIONS Our results strongly indicate a promising potential of nanotopographic features of these surfaces in supporting pivotal neuronal differentiation processes

Proteomic profile confirms nanostructure-induced neuritogenesis and reflects alterations of the mechanotransductive processes in PC12 cells

BACKGROUND-AIM Cells are competent to perceive biophysical signals of their microenvironment and to convert them into biochemical responses. These signals comprise the microenvironmental nanotopography and in fact cells can sense surface differences on a nanoscopic level that can have a strong impact on cellular mechanics and on cell\u2019s behavior. In this work we tried to understand how nanoroughness of zirconia surfaces, produced by Supersonic Cluster Beam Deposition using a Pulsed Microplasma Cluster Source (PMCS-SCBD) technique, can guide cellular activities in the context of neuronal differentiation. The potential of nanostructured surfaces in inducing neurogenesis would have enormous clinical relevance regarding cell replacement strategies for neurodegenerative diseases. METHODS We performed a label-free shotgun proteomic analysis to compare the proteome of PC12 cells grown on neuritogenesis-inducing zirconia nanostructure (nrZr) with the one of cells grown on flat zirconia (flZr) and Poly-LLysine (PLL) in the presence of Nerve Growth Factor (NGF). An Anova test (FDR 0.05) was carried out to identify proteins differentially expressed among the different conditions. We focused only on the comparison between cells grown on nrZr 15 nm rms and on flZr in order to better understand the effect of the surface nanotopography. Proteins were considered differentially expressed if they were present only in flZr or nrZr 15 nm rms or showed significant t-test difference (Post hoc Bonferroni test p value = 0.0167). RESULTS 52 proteins were up-regulated or present only in cells grown on nrZr 15 nm rms, while 54 proteins were down-regulated in cells grown on nrZr 15 nm rms or were present only in cells grown on flZr. Among these proteins, several of them reflect the differentiation processes induced by the nanostructure. We discovered that most of the proteins are involved in adhesome and/or cytoskeletal organisation and that their up- or down regulation is in line with their functions in neuronal differentiation processes and/or neuritogenesis. CONCLUSIONS Altogether, our results have unravelled some very interesting candidates for more detailed future analysis of mechanotransductive processes induced by nanostructured surfaces

Appassionati Dissodatori. Storia e storiografia della televisione in Italia. Studi in onore di Aldo Grasso

Questo volume nasce come un omaggio al lavoro di Aldo Grasso che, dagli anni Settanta, attraverso l\u2019attivit\ue0 di ricerca e di didattica presso l\u2019Universit\ue0 Cattolica del Sacro Cuore di Milano, ma anche \u2013 dagli anni Novanta \u2013 con il quotidiano \u2018corpo a corpo\u2019 coi programmi \u2013 \uabi testi\ubb \u2013 della televisione italiana, e i suoi protagonisti, ha insegnato ai suoi allievi, e al pi\uf9 vasto pubblico raggiunto dalle pagine del \uabCorriere della Sera\ubb, un approccio critico e contemporaneamente appassionato al medium saldamente centrale nella cultura italiana della seconda met\ue0 del Novecento e, ancora, del nuovo secolo. Il volume si focalizza su alcune questioni rilevanti: in che modo \ue8 possibile riflettere criticamente e fare ricerca sul ruolo di un mezzo tanto immediato e popolare quanto complesso nelle sue articolazioni e rilevante dal punto di vista culturale e sociale? Quali prospettive pu\uf2 adottare lo studio storiografico della televisione, e a quali fonti deve ricorrere