TeatroNatura® in Feudozzo

O Thiasos TeatroNatura® answered the invitation to the Cammino Lter in Feudozzo, formulating a contribution that was artistic, educational and of common thought with which to meet a community of researchers from different disciplines interested in ecology and complexity. In researching new learning processes, epistemological models, and ways of interpreting and communicating a reality that is to be integrated with scientific learning models and transform their orientations, the perceptive, imaginative and empathic relationships with one's living body and the surrounding living place becomes of a crucial importance, especially in a natural context. The proposed experience points out the living quality of participation in the theater workshop and the encounter with the aesthetic dimension through a performance by TeatroNatura® with traditional polyphonic songs from rural cultures and the oral narration of an ancient myth. The face-to-face exchange of resonances and elements of different disciplines for a shared creation of knowledge becomes beneficial for the participants, and is an integral part of the theoretical vitality of the processes that unfolded during the meeting. The hypothesis, to be examined more thoroughly, is that all these aspects can regenerate contact with the senses, the body’s memories, cognitive openness and flexibility, the poetic relationship with the archetypal figurations that live in Western culture, and in doing so start radically questioning the hegemonic patriarchal mind that condition all fields of recognized knowledge, which is fully shifted towards its intellectual, mercified and competitive aspect, to the detriment of the affective, sensorial and emphatic side. The centrality of scientific thought is not denied, but the aim is to reconnect it with the mysterious and vulnerable depth from which every human desire for knowledge comes. There is no real knowledge without any contact with that reality. To establish a new culture, we need to find a new way to make a successful alliance between science and artistic thinking and to open up to lived wisdoms, close to the health of the land of non-Western people. In this paper we consider the embedded knowledge of performative art as a contribution to common research: understanding how to reach a new ethics, the need for which has been inexorably highlighted by the current pandemic, centered on personal health, that of living beings and the planet we inhabit. Our intention is not to internalize moral dictates or uplifting intentions, but to challenge all forms of knowledge in order to find tools, languages, poetics and practices that work together to effectively take root – those forms first of all - in the care of the common good, namely in the human journey through wisdom for a generalized and personalized development of the art of living. We must learn to consider the presence at various levels of Nature in every cognitive act as essential and the hypothesis of an enlargement of the identity of humans to living beings, to learn to build an alliance among the living beings which is now indispensable. One of the questions that can no longer be postponed is on the tragic existential separation between knowing and being, both on an individual and cultural level, which prevents our civilization from acting for the common good

Interactions of Graphene Oxide and Few-Layer Graphene with the Blood−Brain Barrier

We thank Dr. Michele Dipalo (Istituto Italiano di Tecnologia, Genova, Italy) for help with Raman measurements. We also thank Ilaria Dallorto, Rossana Ciancio, Diego Moruzzo, and Arta Mehilli for administrative and technical help. The project has received funding from the European Union’s Horizon 2020 Research and Innovation Programme under Grant Agreement No. 881603 Graphene Flagship Core3 (to F.B.), the Italian Ministry of Foreign Affairs and International Cooperation (Grant Agreement No. MAE00694702021-05-20 to F.B.), and IRCCS Ospedale Policlinico San Martino, Genova, Italy (Ricerca Corrente and “5x1000” to F.B and V.C.).Materials and methods and additional figures on materials characterization, in vitro experiments, imaging and mass spectrometry analysis. The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.nano- lett.3c00377. Materials and methods and additional figures on materials characterization, in vitro experiments, imaging and mass spectrometry analysis (PDF) https://pubs.acs.org/doi/suppl/10.1021/acs.nanolett.3c00377/suppl_file/nl3c00377_si_001.pdf Supplementary File P1: Full list of quantified proteins (XLSX) Supplementary File P2: Full sets of altered proteins (XLSX) Supplementary File P3: Full sets of altered proteins (XLSX)Thanks to their biocompatibility and high cargo capability, graphene-based materials (GRMs) might represent an ideal brain delivery system. The capability of GRMs to reach the brain has mainly been investigated in vivo and has highlighted some controversy. Herein, we employed two in vitro BBB models of increasing complexity to investigate the bionano interactions with graphene oxide (GO) and few-layer graphene (FLG): a 2D murine Transwell model, followed by a 3D human multicellular assembloid, to mimic the complexity of the in vivo architecture and intercellular crosstalk. We developed specific methodologies to assess the translocation of GO and FLG in a label-free fashion and a platform applicable to any nanomaterial. Overall, our results show good biocompatibility of the two GRMs, which did not impact the integrity and functionality of the barrier. Sufficiently dispersed subpopulations of GO and FLG were actively uptaken by endothelial cells; however, the translocation was identified as a rare event.European Union's Horizon 2020 Research and Innovation Programme 881603Ministry of Foreign Affairs and International Cooperation (Italy) MAE00694702021-05-20IRCCS Ospedale Policlinico San Martino, Genova, Ital

Isobaric Labeling Proteomics Allows a High-Throughput Investigation of Protein Corona Orientation

The formation of the biomolecular corona represents a crucial factor in controlling the biological interactions and trafficking of nanomaterials. In this context, the availability of key epitopes exposed on the surface of the corona, and able to engage the biological machinery, is important to define the biological fate of the material. While the full biomolecular corona composition can be investigated by conventional bottom-up proteomics, the assessment of the spatial orientation of proteins in the corona in a high-throughput fashion is still challenging. In this work, we show that labeling corona proteins with isobaric tags in their native conditions and analyzing the MS/MS spectra of tryptic peptides allow an easy and high-throughput assessment of the inner/outer orientation of the corresponding proteins in the original corona. We put our results in the context of what is currently known of the protein corona of graphene-based nanomaterials. Our conclusions are in line with previous data and were confirmed by in silico calculations.European Commission 754446 881603European Union's Horizon 2020 under the Marie SklodowskaCurie Action-COFUND Athenea3i 75444

Neuronal hyperactivity causes Na+/H+ exchanger-induced extracellular acidification at active synapses

Extracellular pH impacts on neuronal activity, which is in turn an important determinant of extracellular H+ concentration. The aim of this study is to describe the spatio-temporal dynamics of extracellular pH at synaptic sites during neuronal hyperexcitability. To address this issue we created ex.E2GFP, a membrane-targeted extracellular ratiometric pH indicator exquisitely sensitive to acidic shifts. By monitoring ex.E2GFP fluorescence in real time in primary cortical neurons we were able to quantify pH fluctuations during network hyperexcitability induced by convulsant drugs or high frequency electrical stimulation. Sustained hyperactivity caused a pH decrease that was reversible upon silencing of neuronal activity and localized to active synapses. This acidic shift was not attributable to the outflow of synaptic vesicle protons into the cleft nor to the activity of membrane-exposed H+-vATPase, but rather to the activity of the Na+/H+-exchanger. Our data demonstrate that extracellular synaptic pH shifts take place during epileptic-like activity of neural cultures, underlying the strict links existing between synaptic activity and synaptic pH. This evidence may contribute to the understanding of the physio-pathological mechanisms associated with hyperexcitability in the epileptic brain

Interfacing Graphene-Based Materials With Neural Cells

The scientific community has witnessed an exponential increase in the applications of graphene and graphene-based materials in a wide range of fields, from engineering to electronics to biotechnologies and biomedical applications. For what concerns neuroscience, the interest raised by these materials is two-fold. On one side, nanosheets made of graphene or graphene derivatives (graphene oxide, or its reduced form) can be used as carriers for drug delivery. Here, an important aspect is to evaluate their toxicity, which strongly depends on flake composition, chemical functionalization and dimensions. On the other side, graphene can be exploited as a substrate for tissue engineering. In this case, conductivity is probably the most relevant amongst the various properties of the different graphene materials, as it may allow to instruct and interrogate neural networks, as well as to drive neural growth and differentiation, which holds a great potential in regenerative medicine. In this review, we try to give a comprehensive view of the accomplishments and new challenges of the field, as well as which in our view are the most exciting directions to take in the immediate future. These include the need to engineer multifunctional nanoparticles (NPs) able to cross the blood-brain-barrier to reach neural cells, and to achieve on-demand delivery of specific drugs. We describe the state-of-the-art in the use of graphene materials to engineer three-dimensional scaffolds to drive neuronal growth and regeneration in vivo, and the possibility of using graphene as a component of hybrid composites/multi-layer organic electronics devices. Last but not least, we address the need of an accurate theoretical modeling of the interface between graphene and biological material, by modeling the interaction of graphene with proteins and cell membranes at the nanoscale, and describing the physical mechanism(s) of charge transfer by which the various graphene materials can influence the excitability and physiology of neural cells

Graphene Oxide Upregulates the Homeostatic Functions of Primary Astrocytes and Modulates Astrocyte-to-Neuron Communication

Graphene-based materials are the focus of intense research efforts to devise novel theranostic strategies for targeting the central nervous system. In this work, we have investigated the consequences of long-term exposure of primary rat astrocytes to pristine graphene (GR) and graphene oxide (GO) flakes. We demonstrate that GR/GO interfere with a variety of intracellular processes as a result of their internalization through the endolysosomal pathway. Graphene-exposed astrocytes acquire a more differentiated morphological phenotype associated with extensive cytoskeletal rearrangements. Profound functional alterations are induced by GO internalization, including the upregulation of inward-rectifying K+ channels and of Na+-dependent glutamate uptake, which are linked to the astrocyte capacity to control the extracellular homeostasis. Interestingly, GO-pretreated astrocytes promote the functional maturation of co-cultured primary neurons by inducing an increase in intrinsic excitability and in the density of GABAergic synapses. The results indicate that graphene nanomaterials profoundly affect astrocyte physiology in vitro with consequences for neuronal network activity. This work supports the view that GO-based materials could be of great interest to address pathologies of the central nervous system associated with astrocyte dysfunctions

Graphene nanoplatelets render poly(3-hydroxybutyrate) a suitable scaffold to promote neuronal network development

The use of composite biomaterials as innovative bio-friendly neuronal interfaces has been poorly developed so far. Smart strategies to target neuro-pathologies are currently exploiting the mixed and complementary characteristics of composite materials to better design future neural interfaces. Here we present a polymer-based scaffold that has been rendered suitable for primary neurons by embedding graphene nanoplatelets (GnP). In particular, the growth, network formation, and functionality of primary neurons on poly(3-hydroxybutyrate) [P(3HB)] polymer supports functionalized with various concentrations of GnP were explored. After growing primary cortical neurons onto the supports for 14 days, all specimens were found to be biocompatible, revealing physiological growth and maturation of the neuronal network. When network functionality was investigated by whole patch-clamp measurements, pure P(3HB) led to changes in the action potential waveform and reduction in firing frequency, resulting in decreased neuronal excitability. However, the addition of GnP to the polymer matrix restored the electrophysiological parameters to physiological values. Interestingly, a low concentration of graphene was able to promote firing activity at a low level of injected current. The results indicate that the P(3HB)/GnP composites show great potential for electrical interfacing with primary neurons to eventually target central nervous system disorders

An Increase in Membrane Cholesterol by Graphene Oxide Disrupts Calcium Homeostasis in Primary Astrocytes

The use of graphene nanomaterials (GNMs) for biomedical applications targeted to the central nervous system is exponentially increasing, although precise information on their effects on brain cells is lacking. In this work, the molecular changes induced in cortical astrocytes by few-layer graphene (FLG) and graphene oxide (GO) flakes are addressed. The results show that exposure to FLG/GO does not affect cell viability or proliferation. However, proteomic and lipidomic analyses unveil alterations in several cellular processes, including intracellular Ca2+ ([Ca2+ ]i ) homeostasis and cholesterol metabolism, which are particularly intense in cells exposed to GO. Indeed, GO exposure impairs spontaneous and evoked astrocyte [Ca2+ ]i signals and induces a marked increase in membrane cholesterol levels. Importantly, cholesterol depletion fully rescues [Ca2+ ]i dynamics in GO-treated cells, indicating a causal relationship between these GO-mediated effects. The results indicate that exposure to GNMs alters intracellular signaling in astrocytes and may impact astrocyte-neuron interactions