Identifying conformational changes with site-directed spin labeling reveals that the GTPase domain of HydF is a molecular switch

[FeFe]-hydrogenases catalyse the reduction of protons to hydrogen at a complex 2Fe[4Fe4S] center called H-cluster. The assembly of this active site is a multistep process involving three proteins, HydE, HydF and HydG. According to the current models, HydF has the key double role of scaffold, upon which the final H-cluster precursor is assembled, and carrier to transfer it to the target hydrogenase. The X-ray structure of HydF indicates that the protein is a homodimer with both monomers carrying two functional domains: a C-terminal FeS cluster-binding domain, where the precursor is assembled, and a N-terminal GTPase domain, whose exact contribution to cluster biogenesis and hydrogenase activation is still elusive. We previously obtained several hints suggesting that the binding of GTP to HydF could be involved in the interactions of this scaffold protein with the other maturases and with the hydrogenase itself. In this work, by means of site directed spin labeling coupled to EPR/PELDOR spectroscopy, we explored the conformational changes induced in a recombinant HydF protein by GTP binding, and provide the first clue that the HydF GTPase domain could be involved in the H-cluster assembly working as a molecular switch similarly to other known small GTPases

8-Chloro-Cyclic AMP and Protein Kinase A I-Selective Cyclic AMP Analogs Inhibit Cancer Cell Growth through Different Mechanisms

Cyclic AMP (cAMP) inhibits the proliferation of several tumor cells. We previously reported an antiproliferative effect of PKA I-selective cAMP analogs (8-PIP-cAMP and 8-HA-cAMP) on two human cancer cell lines of different origin. 8-Cl-cAMP, another cAMP analog with known antiproliferative properties, has been investigated as a potential anticancer drug. Here, we compared the antiproliferative effect of 8-Cl-cAMP and the PKA I-selective cAMP analogs in three human cancer cell lines (ARO, NPA and WRO). 8-Cl-cAMP and the PKA I-selective cAMP analogs had similarly potent antiproliferative effects on the BRAF-positive ARO and NPA cells, but not on the BRAF-negative WRO cells, in which only 8-Cl-cAMP consistently inhibited cell growth. While treatment with the PKA I-selective cAMP analogs was associated with growth arrest, 8-Cl-cAMP induced apoptosis. To further investigate the actions of 8-Cl-cAMP and the PKA I-selective cAMP analogs, we analyzed their effects on signaling pathways involved in cell proliferation and apoptosis. Interestingly, the PKA I-selective cAMP analogs, but not 8-Cl-cAMP, inhibited ERK phosphorylation, whereas 8-Cl-cAMP alone induced a progressive phosphorylation of the p38 mitogen-activated protein kinase (MAPK), via activation of AMPK by its metabolite 8-Cl-adenosine. Importantly, the pro-apoptotic effect of 8-Cl-cAMP could be largely prevented by pharmacological inhibition of the p38 MAPK. Altogether, these data suggest that 8-Cl-cAMP and the PKA I-selective cAMP analogs, though of comparable antiproliferative potency, act through different mechanisms. PKA I-selective cAMP analogs induce growth arrest in cells carrying the BRAF oncogene, whereas 8-Cl-cAMP induce apoptosis, apparently through activation of the p38 MAPK pathway

Palmitoylethanolamide inhibits rMCP-5 expression by regulating MITF activation in rat chronic granulomatous inflammation

Chronic inflammation, a condition frequently associated with several pathologies, is characterized by angiogenic and fibrogenic responses that may account for the development of granulomatous tissue. We previously demonstrated that the chymase, rat mast cell protease-5 (rMCP-5), exhibits pro-inflammatory and pro-angiogenic properties in a model of chronic inflammation sustained by mast cells (MCs), granuloma induced by the subcutaneous carrageenan-soaked sponge implant in rat. In this study, we investigated the effects of palmitoylethanolamide (PEA), an anti-inflammatory and analgesic endogenous compound, on rMCP-5 mRNA expression and Microphtalmia-associated Transcription Factor (MITF) activation in the same model of chronic inflammation. The levels of rMCP-5 mRNA were detected using semi-quantitative RT-PCR; the protein expression of chymase and extracellular signal-regulated kinases (ERK) were analyzed by western blot; MITF/DNA binding activity and MITF phosphorylation were assessed by electrophoretic mobility shift assay (EMSA) and immunoprecipitation, respectively. The administration of PEA (200, 400 and 800 µg/ml) significantly decreased rMCP-5 mRNA and chymase protein expression induced by λ-carrageenan. These effects were associated with a significant decrease of MITF/DNA binding activity and phosphorylated MITF as well as phosphorylated ERK levels. In conclusion, our results, showing the ability of PEA to inhibit MITF activation and chymase expression in granulomatous tissue, may yield new insights into the understanding of the signaling pathways leading to MITF activation controlled by PEA

Use of benzylglycinamide by a HIV-seropositive polysubstance user: : the changing pattern of novel psychoactive substance use among youths

This document is the Accepted Manuscript of the following article: Matteo Caloro, et al, ‘Use of benzylglycinamide by a HIV-seropositive polysubstance user: The changing pattern of novel psychoactive substance use among youths’, Addictive Behaviors, Vol. 60, pp. 53-57, September 2016. The Version of Record is available online at doi: https://doi.org/10.1016/j.addbeh.2016.03.032. © 2016 Elsevier Ltd. All rights reserved.A 24-year old woman with multisubstance use since the age of 13, including opioids and cocaine, and long-standing HIV/HCV seropositivity status, presented with psychosis, agitation, and insomnia at the emergency department of a university hospital. She had been abusive and physically aggressive frequently without specific reasons and was involved in criminal legal cases. She was hospitalized twice. During her first hospital stay she experienced a brief episode of detachment from her environment, similar to episodes reportedly suffered at home. Psychosis had developed following heavy polysubstance abuse. Her mother provided sachets containing benzylglycinamide, a substance with no known psychotropic effects, which were also present in the patient's urine. She was occasionally positive for cannabinoids. She used to buy various novel psychoactive substances (NPSs) from the internet and used experimentally various substances freely made available to her by drug suppliers/dealers. She was unable to explain clearly why she was taking any of the identified NPS. She stated she was taking benzylglycinamide to calm her when smoking synthetic cannabinoids. While it appears that benzylglycinamide is not likely to constitute a novel drug of abuse, her polysubstance use exemplifies trends in NPS use patterns among the youths in the Western world and should alert mental health workers as to the possible dangers of such behavior and its reflection on social behavior and psychopathology.Peer reviewedFinal Accepted Versio

ICT tools for enhancing sustainable water management in rural environments

Numerical models are relevant tools to achieve the proper application of the Water Framework Directive (WFD, EU 2000). Their major advantage is to foster a full characterization of the involved flow terms and contaminant transport pathways. Thanks to their predictive function, numerical models can help also to address planning and management activities. Many hydrological codes developed so far have faced the problem of tackling multiscale territorial planning (e.g., Bergez et al. 2012). We introduce here the GIS-integrated FREEWAT platform aimed at providing a unique modeling environment to simulate multiple hydrological processes, with a focus on the sustainable management of conjunctive use of surface- and ground-water resources in rural environments. FREEWAT (FREE and open source software tools for WATer resource management; Rossetto et al., 2015) is an EU HORIZON 2020 project, whose main goal is to simplify the application of EU water-related Directives. It aims at integrating a simulation platform in a Geographic Information System (GIS), coupling the power of GIS geo-processing and post-processing tools in spatial data analysis to that of simulation codes. The FREEWAT platform is being developed within the QGIS free open source software package and fosters the simulation of the whole hydrological cycle using open source numerical codes mainly belonging to the USGS MODFLOW family

Allocazione delle risorse per l’attuazione del “Programma dei Lavori Pubblici" per il Triennio 2020-2022 ed “Elenco dei Lavori” 2020.

Per l’attuazione degli interventi edilizi dell’Istituto relativamente al triennio 2020-2022 si è predisposto il quadro di dettaglio delle risorse che sono state rese disponibili per tale scopo, con riferimento alla opportuna allocazione delle medesime nel relativo Bilancio di previsione per l’annualità 2021. Il rapporto a tal fine preparato dalla struttura tecnica denominata “Tavolo Tecnico Permanente per il Patrimonio Immobiliare e i Lavori Pubblici”, è stato redatto sulla scorta degli approfondimenti tecnici ed amministrativi compiuti in fase di istruttoria sulle richieste formulate dalle Strutture di ricerca ubicate su tutto il territorio nazionale, costituenti il quadro generale delle esigenze

Relazione tecnica e tabella contenente gli importi, le coordinate di bilancio di provenienza e di destinazione nonché le “Strutture di Ricerca” destinatarie dei fondi di cui allo “Elenco dei lavori” per l’anno 2020 del “Programma triennale dei lavori pubblici 2020-2022” approvato con Delibera del 31 marzo 2021, numero 14.

Il presente rapporto tecnico si pone l’obiettivo di individuare gli importi da destinare alle “Strutture di ricerca” di cui allo “elenco dei lavori” per l’anno 2020 del “Programma triennale lavori pubblici 2020-2022” approvato con delibera del 31 marzo 2021, numero 14 con le relative coordinate di bilancio sia di provenienza che di destinazione secondo quanto previsto con riferimento al Decreto 25 gennaio 2019, “Aggiornamento dell’allegato 1 del Decreto del Presidente della Repubblica 4 ottobre 2013, numero 132 “Piano dei conti integrato”, ai sensi dell’articolo 5 del medesimo D.P.R. e del comma 4, articolo 4 del Decreto Legislativo 31 maggio 2011, numero 91”

I Lavori Pubblici nello “Istituto Nazionale di Astrofisica” nel triennio 2020 - 2022. Adozione dello schema di “Programma Triennale Lavori Pubblici dello "Istituto Nazionale di Astrofisica" Triennio 2020-2022, "Elenco dei Lavori” per l’anno 2020, ed “Elenco degli interventi sotto 100 mila euro”.

Il rapporto tecnico contiene approfondimenti ai fini della adozione del “Programma Triennale dei Lavori Pubblici 2020 – 2022”, nella sua articolazione annuale e triennale, volto alla riqualificazione, gestione, recupero, attraverso una attenta e corretta programmazione, dell’ingente patrimonio immobiliare dell’Ente, caratterizzato prevalentemente da strutture di grande pregio storico, artistico, scientifico e paesaggistico. Il documento ha inoltre la finalità di definire gli interventi da inserire nello “Schema di Programma Triennale dei Lavori Pubblici 2020-2022", in particolare nell’“Elenco dei Lavori” per l’anno 2020, selezionati in considerazione delle priorità stabilite per Legge in relazione alle disponibilità economiche autonome delle Strutture di Ricerca e della proposta di assegnazione dei fondi destinati ad interventi di edilizia nel Bilancio dell’Ente avanzata dal “Tavolo Tecnico Permanente per il Patrimonio Immobiliare e i Lavori Pubblici”

Application of Patients Reported Outcome measures in Cochlear Implant patients: implications for the design of specific rehabilitation programs

Introduction: Cochlear implants (CI) had been developed to enable a satisfying verbal communication, music experiences have remained in the background of research and development and consequently many CI users are dissatisfied by the music they listen to. Nonetheless, concise indications for clinicians to test music abilities and prescribe rehabilitation programs are still lacking. The main aim of the present study was to test the utility of the application of two different Patient Reporting Outcomes (PRO) measures in a group of CI users. A secondary objective was to identify items capable of driving the indication and design specific music rehabilitation programs for CI patients. Material and Methods: A consecutive series of 73 CI patients referred to the Audiology Unit - University of Padova - was enrolled from November 2021 to May 2022 and evaluated with audiological battery test and PRO measures: Musica e Qualità della Vita (MUSQUAV) and Nijmegen Cochlear Implant Questionnaire (NCIQ) Italian version. Results: The Reliability analysis showed good consistency between the different PRO measures, (Cronbach alpha = 0.873). After accounting for epidemiological and clinical variables, PRO measures showed a correlation with audiological outcomes in only one case (rho=-0.170 for NCIQ-T with CI-Pure Tone Average. A willingness for musical rehabilitation was present in 63% of patients, (Rehab Factor, mean value of 0.791±0.675). Conclusions: we support the role of the application of MUSQUAV and NCIQ to improve the clinical and audiological evaluation of CI patients. Moreover, we proposed a derivative item, called Rehab Factor, which could be used in clinical practice and future studies to clarify the indication and priority of specific music rehabilitation programs