Synaptic P2X7 and oxygen/glucose deprivation in organotypic hippocampal cultures.

The P2X7 receptor for extracellular ATP is the main candidate, among P2 receptors, inducing cell death in the immune system. Here, we demonstrate the direct participation of this receptor to cell damage induced by oxygen/glucose deprivation, in the ex vivo model of organotypic hippocampal cultures. By pharmacological and immunological approaches, we show that P2X7 is rapidly and transiently up regulated in hippocampal areas eliciting metabolism impairment. Moreover, the P2 antagonists 2′,3′,-dialdehyde ATP and reactive blue 2 prevent both up regulation of this receptor and hypoxic/hypoglycemic damage. By confocal laser microscopy, we show that P2X7 is present at the synaptic level of fibers extending from the CA1–2 pyramidal cell layer throughout the strata oriens and radiatum, but absent on oligodendrocytes, astrocytes or neuronal cell bodies. Colocalization of P2X7 is obtained with neurofilament-L protein and with synaptophysin, not with myelin basic protein, glial fibrillary acidic protein or a marker for neuronal nuclei. P2X7 up regulation and diffuse cellular damage are also induced by 3′-O-(4-benzoyl) benzoyl-ATP, an agonist selective but not exclusive for P2X7. In summary, our study demonstrates that P2X7 not only directly participates to the hypoxic/hypoglycemic process, but also owns specific phenotypic localization. We do not exclude that it might serve as a sensor of dysregulated neuronal activity and ATP release, both occurring during oxygen/glucose deprivation

EVENTI: un’indagine sulla resilienza delle istituzioni musicali della Svizzera italiana in tempi di pandemia

Nel quadro di una più ampia riflessione sulle ragioni della persistente segmentazione del pubblico degli eventi culturali, questo contributo nasce da un progetto strategico di ricerca sostenuto dalla Scuola Universitaria Professionale della Svizzera Italiana (SUPSI) con il quale si intende indagare come le istituzioni culturali e artistiche attive nella Svizzera italiana stiano affrontando la fase seguita al lock down imposto dalla pandemia di COVID-19. Un team multidisciplinare, in collaborazione con le principali istituzioni culturali attive sul territorio, intende identificare i fattori, le variabili e le soluzioni imposte dalla nuova situazione, allo scopo di mettere a disposizione dei differenti attori strumenti di conoscenza e di lavoro aggiornati. Nella prima fase del progetto, i rappresentanti di cinque enti attivi nella produzione di eventi di musica classica e jazz sono stati intervistati allo scopo di raccogliere informazioni sulla maniera in cui è stato vissuto e gestito il periodo tra la primavera del 2020 e quella del 2021. I primi risultati indicano un paradossale aumento del lavoro organizzativo e degli oneri amministrativi causato dalle procedure per la richiesta di fondi compensativi e dalla necessità di aggiornare costantemente l’offerta. Allo stesso tempo si registra una forte accelerazione nell’uso delle tecnologie digitali e dei social network. Tuttavia, l’ibridazione dei modelli performativi dovuta a questa accelerazione sembra rispondere ad una funzione palliativa, piuttosto che all’esigenza sentita di ridefinire i tradizionali modelli di consumo musicale.Within a broader reflection on the reasons for the persistent segmentation of the audience of cultural events, this contribution stems from a strategic research project supported by the University of Applied Sciences and Arts of Southern Switzerland (SUPSI). The project investigates how cultural and artistic institutions active in Italian-speaking Switzerland have been coping with the constraints imposed by the COVID-19 pandemic. A multidisciplinary team, in collaboration with the main cultural stakeholder in the region, intends to identify the factors, variables and solutions imposed by the new situation, and provide the various actors with up-to-date knowledge and working tools. Representatives of five organisations active in the production of classical and jazz music events were interviewed in order to collect information on how the period between spring 2020 and spring 2021 was experienced and managed. First results indicate a paradoxical increase in organisational work and administrative burdens caused by the procedures for applying for compensatory funds and the need to constantly update the offer. At the same time, there is a strong acceleration in the use of digital technologies and social networks. However, the hybridisation of performance models related to this acceleration seems to respond to a palliative function, rather than to the felt need to redefine traditional patterns of music consumption.Nel quadro di una più ampia riflessione sulle ragioni della persistente segmentazione del pubblico degli eventi culturali, questo contributo nasce da un progetto strategico di ricerca sostenuto dalla Scuola Universitaria Professionale della Svizzera Italiana (SUPSI) con il quale si intende indagare come le istituzioni culturali e artistiche attive nella Svizzera italiana stiano affrontando la fase seguita al lock down imposto dalla pandemia di COVID-19. Un team multidisciplinare, in collaborazione con le principali istituzioni culturali attive sul territorio, intende identificare i fattori, le variabili e le soluzioni imposte dalla nuova situazione, allo scopo di mettere a disposizione dei differenti attori strumenti di conoscenza e di lavoro aggiornati. Nella prima fase del progetto, i rappresentanti di cinque enti attivi nella produzione di eventi di musica classica e jazz sono stati intervistati allo scopo di raccogliere informazioni sulla maniera in cui è stato vissuto e gestito il periodo tra la primavera del 2020 e quella del 2021. I primi risultati indicano un paradossale aumento del lavoro organizzativo e degli oneri amministrativi causato dalle procedure per la richiesta di fondi compensativi e dalla necessità di aggiornare costantemente l’offerta. Allo stesso tempo si registra una forte accelerazione nell’uso delle tecnologie digitali e dei social network. Tuttavia, l’ibridazione dei modelli performativi dovuta a questa accelerazione sembra rispondere ad una funzione palliativa, piuttosto che all’esigenza sentita di ridefinire i tradizionali modelli di consumo musicale.Nel quadro di una più ampia riflessione sulle ragioni della persistente segmentazione del pubblico degli eventi culturali, questo contributo nasce da un progetto strategico di ricerca sostenuto dalla Scuola Universitaria Professionale della Svizzera Italiana (SUPSI) con il quale si intende indagare come le istituzioni culturali e artistiche attive nella Svizzera italiana stiano affrontando la fase seguita al lock down imposto dalla pandemia di COVID-19. Un team multidisciplinare, in collaborazione con le principali istituzioni culturali attive sul territorio, intende identificare i fattori, le variabili e le soluzioni imposte dalla nuova situazione, allo scopo di mettere a disposizione dei differenti attori strumenti di conoscenza e di lavoro aggiornati. Nella prima fase del progetto, i rappresentanti di cinque enti attivi nella produzione di eventi di musica classica e jazz sono stati intervistati allo scopo di raccogliere informazioni sulla maniera in cui è stato vissuto e gestito il periodo tra la primavera del 2020 e quella del 2021. I primi risultati indicano un paradossale aumento del lavoro organizzativo e degli oneri amministrativi causato dalle procedure per la richiesta di fondi compensativi e dalla necessità di aggiornare costantemente l’offerta. Allo stesso tempo si registra una forte accelerazione nell’uso delle tecnologie digitali e dei social network. Tuttavia, l’ibridazione dei modelli performativi dovuta a questa accelerazione sembra rispondere ad una funzione palliativa, piuttosto che all’esigenza sentita di ridefinire i tradizionali modelli di consumo musicale

Synergistic inhibition of the Hedgehog pathway by newly designed Smo and Gli antagonists bearing the isoflavone scaffold

Aberrant activation of the Hedgehog (Hh) pathway is responsible for the onset and progression of several malignancies. Small molecules able to block the pathway at the upstream receptor Smoothened (Smo) or the downstream effector Gli1 have thus emerged recently as valuable anticancer agents. Here, we have designed, synthesized, and tested new Hh inhibitors taking advantage by the highly versatile and privileged isoflavone scaffold. The introduction of specific substitutions on the isoflavone's ring B allowed the identification of molecules targeting preferentially Smo or Gli1. Biological assays coupled with molecular modeling corroborated the design strategy, and provided new insights into the mechanism of action of these molecules. The combined administration of two different isoflavones behaving as Smo and Gli antagonists, respectively, in primary medulloblastoma (MB) cells highlighted the synergistic effects of these agents, thus paving the way to further and innovative strategies for the pharmacological inhibition of Hh signaling

Inhibition of Hedgehog-dependent tumors and cancer stem cells by a newly identified naturally occurring chemotype

Hedgehog (Hh) inhibitors have emerged as valid tools in the treatment of a wide range of cancers. Indeed, aberrant activation of the Hh pathway occurring either by ligand-dependent or -independent mechanisms is a key driver in tumorigenesis. The smoothened (Smo) receptor is one of the main upstream transducers of the Hh signaling and is a validated target for the development of anticancer compounds, as underlined by the FDA-approved Smo antagonist Vismodegib (GDC-0449/Erivedge) for the treatment of basal cell carcinoma. However, Smo mutations that confer constitutive activity and drug resistance have emerged during treatment with Vismodegib. For this reason, the development of new effective Hh inhibitors represents a major challenge for cancer therapy. Natural products have always represented a unique source of lead structures in drug discovery, and in recent years have been used to modulate the Hh pathway at multiple levels. Here, starting from an in house library of natural compounds and their derivatives, we discovered novel chemotypes of Hh inhibitors by mean of virtual screening against the crystallographic structure of Smo. Hh functional based assay identified the chalcone derivative 12 as the most effective Hh inhibitor within the test set. The chalcone 12 binds the Smo receptor and promotes the displacement of Bodipy-Cyclopamine in both Smo WT and drug-resistant Smo mutant. Our molecule stands as a promising Smo antagonist able to specifically impair the growth of Hh-dependent tumor cells in vitro and in vivo and medulloblastoma stem-like cells and potentially overcome the associated drug resistance

TiO2-Ag-NP adhesive photocatalytic films able to disinfect living indoor spaces with a straightforward approach

TiO2-Ag doped nanoparticulate (TiO2-Ag-NP) adhesive photocatalytic films were used to assess the ability in dropping down the burden of indoor microbial particles. The application of an easy-to use photocatalytic adhesive film to cleanse indoor living spaces from microbial pollution, represents a novelty in the field of photocatalytic devices. Reduction was attained by photocatalysis in selected spaces, usually with overcrowding (≥ 3 individuals) in the common working daily hours, and upon indoor microclimate monitoring. TiO2-Ag doped nanoparticulate (TiO2-Ag-NP) adhesive photocatalytic films were applied within five types of living spaces, including schools and job places. The microbial pollution was assessed at time 0 (far from routine clean, ≥ 9 h) and throughout 2-4 weeks following the photocatalyst application by relative light unit (RLU) luminometry and microbial indirect assessment (colony forming units per cubic meter, CFU/m3). TiO2-Ag-NP photocatalyst reduced RLU and CFU/m3 by rates higher than 70% leading to RLU ≤ 20 and microbial presence ≤ 35 CFU/m3. The described TiO2-Ag-NP is able to reduce microbial pollution to the lowest RLU threshold (≤ 20) within 60 min in open daylight in a standardized test room of 100 m2. The correlation between RLU and CFU/m3 was positive (r = 0.5545, p < 0.05), assessing that the microbial reduction of indoor areas by the TiO2-Ag-NP adhesive film was real. Titania photocatalysts represent promising tools to ensure air cleaning and sanitization in living indoor microclimates with a low cost, feasible and straightforward approach. This approach represents an easy to handle, cost effective, feasible and efficacious approach to reduce microbial pollution in indoor spaces, by simply attaching a TiO2-Ag-NP adhesive film on the wall

Analysis of the root morphology of European anterior teeth

Aim of this study was to investigate the gross anatomy of the root of European anterior teeth. A review of the dental literature shows that in the past the root morphology was investigated from the inner pulp chamber for endodontic therapies. In order to be admitted to the study, the teeth had to be undamaged. Each tooth was identified by a serial number and gauged by a millimeter tape (for the root length), a goniometer (for the root angle), and a millimeter gauge (for the root diameter). Furthermore, a statistical elaboration of the data was performed to underline the shape variations of the surface around the different sides of the root. At the end of the analysis, 12 parameters for each single-root tooth were described. The study highlights significant differences (p<0.01) only in two teeth of the maxillary arch (central incisor and canine) and in one tooth of the mandibular arch (central incisor). In both cases, the observed differences may be due to the sinuosity of the cement-enamel line. The Tables for each measured param- eter were obtained for all examined classes of teeth, but a comparison with literature data was possible only for the “root length” parameter. This study can be considered innovative for the absence, in the scientific literature, of a statistical analysis of all parameters with the exception of the “root length”. Moreover, it gives a detailed updating of the data relative to the European population creating a useful tool as well for surgical interventions during periodontal therapy (for example in the choice of the right ultrasonic handpiece) as for new CAD/CAM assisted implant manufacturing techniques

Identification of an L-Rhamnose Synthetic Pathway in Two Nucleocytoplasmic Large DNA Viruses

Nucleocytoplasmic large DNA viruses (NCLDVs) are characterized by large genomes that often encode proteins not commonly found in viruses. Two species in this group are Acanthocystis turfacea chlorella virus 1 (ATCV-1) (family Phycodnaviridae, genus Chlorovirus) and Acanthamoeba polyphaga mimivirus (family Mimiviridae), commonly known as mimivirus. ATCV-1 and other chlorovirus members encode enzymes involved in the synthesis and glycosylation of their structural proteins. In this study, we identified and characterized three enzymes responsible for the synthesis of the sugar L-rhamnose: two UDP-D-glucose 4,6-dehydratases (UGDs) encoded by ATCV-1 and mimivirus and a bifunctional UDP-4-keto-6-deoxy-D-glucose epimerase/reductase (UGER) from mimivirus. Phylogenetic analysis indicated that ATCV-1 probably acquired its UGD gene via a recent horizontal gene transfer (HGT) from a green algal host, while an earlier HGT event involving the complete pathway (UGD and UGER) probably occurred between a protozoan ancestor and mimivirus. While ATCV-1 lacks an epimerase/reductase gene, its Chlorella host may encode this enzyme. Both UGDs and UGER are expressed as late genes, which is consistent with their role in posttranslational modification of capsid proteins. The data in this study provide additional support for the hypothesis that chloroviruses, and maybe mimivirus, encode most, if not all, of the glycosylation machinery involved in the synthesis of specific glycan structures essential for virus replication and infection

Detection and Quantitative Analysis of Two Independent Binding Modes of a Small Ligand Responsible for DC-SIGN Clustering

DC-SIGN (dendritic cell-specific ICAM-3 grabbing non-integrin) is a C-type lectin receptor (CLRs) present, mainly in dendritic cells (DCs), as one of the major pattern recognition receptors (PRRs). This receptor has a relevant role in viral infection processes. Recent approaches aiming to block DC-SIGN have been presented as attractive anti-HIV strategies. DC-SIGN binds mannose or fucose-containing carbohydrates from viral proteins such as the HIV envelope glycoprotein gp120. We have previously demonstrated that multivalent dendrons bearing multiple copies of glycomimetic ligands were able to inhibit DC-SIGN-dependent HIV infection in cervical explant models. Optimization of glycomimetic ligands requires detailed characterization and analysis of their binding modes because they notably influence binding affinities. In a previous study we characterized the binding mode of DC-SIGN with ligand 1, which shows a single binding mode as demonstrated by NMR and X-ray crystallography. In this work we report the binding studies of DC-SIGN with pseudotrisaccharide 2, which has a larger affinity. Their binding was analysed by TR-NOESY and STD NMR experiments, combined with the CORCEMA-ST protocol and molecular modelling. These studies demonstrate that in solution the complex cannot be explained by a single binding mode. We describe the ensemble of ligand bound modes that best fit the experimental data and explain the higher inhibition values found for ligand

Residual tumor micro-foci and overwhelming regulatory T lymphocyte infiltration are the causes of bladder cancer recurrence

Bladder cancer has an unexplained, high recurrence rate. Causes of recurrence might include the presence of sporadic tumor micro-foci in the residual urothelial tissue after surgery associated with an inverted ratio between intratumoral effector and regulatory T cell subsets. Hence, surgical specimens of both tumors and autologous, macroscopically/histologically free-of-tumor tissues were collected from 28 and 20 patients affected by bladder or renal cancer, respectively. The frequencies of effector (IFN?+ and IL17+ T cells) and regulatory (CD4+CD25hiCD127lo and CD8+CD28-CD127loCD39+ Treg) T cell subpopulations among tumor infiltrating lymphocytes were analyzed by immunofluorescence, while the gene expression of MAGE-A1 and MAGE-A2 tumor-associated antigens was studied by RT-PCR. The results show that both the T cell infiltrate and the frequency of MAGE-A1/A2 gene expression were comparable in tumors and in autologous free-of-tumor tissues in bladder cancer, while the autologous free-of-tumor renal tissues showed reduced T cell infiltrate and frequency of MAGE gene expression as compared to the autologous tumors. Importantly, the intra-tumor T effector/Treg cell ratio was consistently <1 in bladder cancer patients (n. 7) who relapsed within two years, while it was always >1 in patients (n. 6) without recurrence (regardless of tumor stage) (P = 0.0006, Odds ratio = 195). These unprecedented findings clarify the pathogenic mechanism of bladder cancer recurrence and suggest that microscopically undetectable micro-foci of tumor may predispose to recurrence when associated with an inverted intratumoral T effector/Treg cell ratio

Age-dependent roles of peroxisomes in the hippocampus of a transgenic mouse model of Alzheimer’s disease

BACKGROUND: Alzheimer's Disease (AD) is a progressive neurodegenerative disease, especially affecting the hippocampus. Impairment of cognitive and memory functions is associated with amyloid beta-peptide-induced oxidative stress and alterations in lipid metabolism. In this scenario, the dual role of peroxisomes in producing and removing ROS, and their function in fatty acids beta-oxidation, may be critical. This work aims to investigating the possible involvement of peroxisomes in AD onset and progression, as studied in a transgenic mouse model, harboring the human Swedish familial AD mutation. We therefore characterized the peroxisomal population in the hippocampus, focusing on early, advanced, and late stages of the disease (3, 6, 9, 12, 18 months of age). Several peroxisome-related markers in transgenic and wild-type hippocampal formation were comparatively studied, by a combined molecular/immunohistochemical/ultrastructural approach. RESULTS: Our results demonstrate early and significant peroxisomal modifications in AD mice, compared to wild-type. Indeed, the peroxisomal membrane protein of 70 kDa and acyl-CoA oxidase 1 are induced at 3 months, possibly reflecting the need for efficient fatty acid beta-oxidation, as a compensatory response to mitochondrial dysfunction. The concomitant presence of oxidative damage markers and the altered expression of antioxidant enzymes argue for early oxidative stress in AD. During physiological and pathological brain aging, important changes in the expression of peroxisome-related proteins, also correlating with ongoing gliosis, occur in the hippocampus. These age- and genotype-based alterations, strongly dependent on the specific marker considered, indicate metabolic and/or numerical remodeling of peroxisomal population. CONCLUSIONS: Overall, our data support functional and biogenetic relationships linking peroxisomes to mitochondria and suggest peroxisomal proteins as biomarkers/therapeutic targets in pre-symptomatic AD