Inter and intra-population variability of the migratory behaviour of a short-distance partial migrant, the Eurasian Stone-curlew Burhinus oedicnemus (Charadriiformes, Burhinidae)

Migratory behaviour in birds shows a remarkable variability at species, population and individual levels. Short-distance migrants often adopt a partial migratory strategy and tend to have a flexible migration schedule that allows a more effective response to extreme environmental variations. Weather seasonality and environmental heterogeneity have been reported as significant factors in the diversification of migratory behaviour for Mediterranean migrants, but relatively few studies investigated the migration patterns of non-passerine birds migrating within the Mediterranean basin. In this study, we investigated the migratory strategy of 40 Eurasian Stone-curlews Burhinus oedicnemus tagged with geolocators and GPS-GSM tags and belonging to continental and Mediterranean populations of the Italian peninsula. The proportion of migrants was higher in continental populations, but we observed a significant variability also within Mediterranean populations. All birds spent the winter within the Mediterranean basin. Continental Stone-curlews departed earlier in spring and later in autumn and covered longer distances than those from Mediterranean areas. The speed of migration did not change between seasons for continental birds, while Mediterranean individuals migrated faster in spring. The likelihood of departure for autumn migration of GPS-tagged birds increased when temperatures were near or below 0 °C suggesting that Stone-curlews tend to delay departure until weather conditions worsen abruptly. As a consequence of global warming in the Mediterranean, the frequency of migratory birds in the considered populations might decrease in the near future. This could affect the distribution of species throughout the year and should be taken into account when targeting conservation measures

Lack of SARS-CoV-2 RNA environmental contamination in a tertiary referral hospital for infectious diseases in Northern Italy

none140noNAnoneColaneri M.; Seminari E.; Piralla A.; Zuccaro V.; Di Filippo A.; Baldanti F.; Bruno R.; Mondelli M.U.; Brunetti E.; Di Matteo A.; Maiocchi L.; Pagnucco L.; Mariani B.; Ludovisi S.; Lissandrin R.; Parisi A.; Sacchi P.; Patruno S.F.A.; Michelone G.; Gulminetti R.; Zanaboni D.; Novati S.; Maserati R.; Orsolini P.; Vecchia M.; Sciarra M.; Asperges E.; Sambo M.; Biscarini S.; Lupi M.; Roda S.; Chiara Pieri T.; Gallazzi I.; Sachs M.; Valsecchi P.; Perlini S.; Alfano C.; Bonzano M.; Briganti F.; Crescenzi G.; Giulia Falchi A.; Guarnone R.; Guglielmana B.; Maggi E.; Martino I.; Pettenazza P.; Pioli di Marco S.; Quaglia F.; Sabena A.; Salinaro F.; Speciale F.; Zunino I.; De Lorenzo M.; Secco G.; Dimitry L.; Cappa G.; Maisak I.; Chiodi B.; Sciarrini M.; Barcella B.; Resta F.; Moroni L.; Vezzoni G.; Scattaglia L.; Boscolo E.; Zattera C.; Michele Fidel T.; Vincenzo C.; Vignaroli D.; Bazzini M.; Iotti G.; Mojoli F.; Belliato M.; Perotti L.; Mongodi S.; Tavazzi G.; Marseglia G.; Licari A.; Brambilla I.; Daniela B.; Antonella B.; Patrizia C.; Giulia C.; Giuditta C.; Marta C.; Rossana D.; Milena F.; Bianca M.; Roberta M.; Enza M.; Stefania P.; Maurizio P.; Elena P.; Antonio P.; Francesca R.; Antonella S.; Maurizio Z.; Guy A.; Laura B.; Ermanna C.; Giuliana C.; Luca D.; Gabriella F.; Gabriella G.; Alessia G.; Viviana L.; Claudia L.; Valentina M.; Simona P.; Marta P.; Alice B.; Giacomo C.; Irene C.; Alfonso C.; Di Martino R.; Di Napoli A.; Alessandro F.; Guglielmo F.; Loretta F.; Federica G.; Alessandra M.; Federica N.; Giacomo R.; Beatrice R.; Maria S.I.; Monica T.; Nepita Edoardo V.; Calvi M.; Tizzoni M.; Nicora C.; Triarico A.; Petronella V.; Marena C.; Muzzi A.; Lago P.; Comandatore F.; Bissignandi G.; Gaiarsa S.; Rettani M.; Bandi C.Colaneri, M.; Seminari, E.; Piralla, A.; Zuccaro, V.; Di Filippo, A.; Baldanti, F.; Bruno, R.; Mondelli, M. U.; Brunetti, E.; Di Matteo, A.; Maiocchi, L.; Pagnucco, L.; Mariani, B.; Ludovisi, S.; Lissandrin, R.; Parisi, A.; Sacchi, P.; Patruno, S. F. A.; Michelone, G.; Gulminetti, R.; Zanaboni, D.; Novati, S.; Maserati, R.; Orsolini, P.; Vecchia, M.; Sciarra, M.; Asperges, E.; Sambo, M.; Biscarini, S.; Lupi, M.; Roda, S.; Chiara Pieri, T.; Gallazzi, I.; Sachs, M.; Valsecchi, P.; Perlini, S.; Alfano, C.; Bonzano, M.; Briganti, F.; Crescenzi, G.; Giulia Falchi, A.; Guarnone, R.; Guglielmana, B.; Maggi, E.; Martino, I.; Pettenazza, P.; Pioli di Marco, S.; Quaglia, F.; Sabena, A.; Salinaro, F.; Speciale, F.; Zunino, I.; De Lorenzo, M.; Secco, G.; Dimitry, L.; Cappa, G.; Maisak, I.; Chiodi, B.; Sciarrini, M.; Barcella, B.; Resta, F.; Moroni, L.; Vezzoni, G.; Scattaglia, L.; Boscolo, E.; Zattera, C.; Michele Fidel, T.; Vincenzo, C.; Vignaroli, D.; Bazzini, M.; Iotti, G.; Mojoli, F.; Belliato, M.; Perotti, L.; Mongodi, S.; Tavazzi, G.; Marseglia, G.; Licari, A.; Brambilla, I.; Daniela, B.; Antonella, B.; Patrizia, C.; Giulia, C.; Giuditta, C.; Marta, C.; D'Alterio, Rossana; Milena, F.; Bianca, M.; Roberta, M.; Enza, M.; Stefania, P.; Maurizio, P.; Elena, P.; Antonio, P.; Francesca, R.; Antonella, S.; Maurizio, Z.; Guy, A.; Laura, B.; Ermanna, C.; Giuliana, C.; Luca, D.; Gabriella, F.; Gabriella, G.; Alessia, G.; Viviana, L.; Meisina, Claudia; Valentina, M.; Simona, P.; Marta, P.; Alice, B.; Giacomo, C.; Irene, C.; Alfonso, C.; Di Martino, R.; Di Napoli, A.; Alessandro, F.; Guglielmo, F.; Loretta, F.; Federica, G.; Albertini, Alessandra; Federica, N.; Giacomo, R.; Beatrice, R.; Maria, S. I.; Monica, T.; Nepita Edoardo, V.; Calvi, M.; Tizzoni, M.; Nicora, C.; Triarico, A.; Petronella, V.; Marena, C.; Muzzi, A.; Lago, P.; Comandatore, F.; Bissignandi, G.; Gaiarsa, S.; Rettani, M.; Bandi, C

Clinical characteristics of coronavirus disease (COVID-19) early findings from a teaching hospital in Pavia, North Italy, 21 to 28 February 2020

We describe clinical characteristics, treatments and outcomes of 44 Caucasian patients with coronavirus disease (COVID-19) at a single hospital in Pavia, Italy, from 21\u201328 February 2020, at the beginning of the outbreak in Europe. Seventeen patients developed severe disease, two died. After a median of 6 days, 14 patients were discharged from hospital. Predictors of lower odds of discharge were age>65 years, antiviral treatment and for severe disease, lactate dehydrogenase >300 mg/dL

Factors affecting migratory and foraging behaviour of a partial migratory species, the Eurasian Stone-curlew Burhinus oedicnemus (Charadriiformes, Burhinidae) during the non-breeding period

Movement is a critical component of animal biology, affecting individual fitness and survival and playing an important role in structuring populations and the functioning of ecosystems. Among the different types of movement, migration and foraging movements play an essential role in bird biology. Both types of movements show great variability at species, population and individual level. For what concern migratory behaviour, short-distance migrants for example often adopt a partial migratory strategy and tend to have a flexible migration schedule that allows a more effective response to extreme environmental variations. Weather seasonality and environmental heterogeneity have been reported as significant factors in the diversification of migratory behaviour for Mediterranean migrants, but relatively few studies investigated the migration patterns of non-passerine birds migrating within this area. Foraging movements occur generally at smaller scale, but like migration they show high variability in patterns that may change between and within individuals even belonging to the same population. External factors such as food supply and habitat structure can influence the variability of foraging behaviour and the agricultural ecosystem, where habitat modifications act at different temporal and spatial scales, is a good setting to study this topic. In Europe, intensive farming practices were reported as the main factors in changing of foraging behaviour in farmland species due to the loss of suitable areas during the breeding period, but relatively few studies investigated the spatial behaviour during the non-breeding period. Understanding the factors that influence animal movements at different spatial and temporal scales is crucial in the study of movement ecology, especially because it can allow us to predict how animals will respond to changes in environmental conditions. In this dissertation, I focused on the factors affecting the spatial behaviour of the Eurasian Stone-curlew Burhinus oedicnemus, a short distance migrant inhabiting the European agricultural landscape, during the non-breeding In chapter 2 I studied the migratory strategy of 40 individuals tagged with geolocators and GPS-GSM tags during the period 2010-2019 and belonging to continental and Mediterranean populations of the Italian peninsula. The proportion of migrants turned out to be higher in continental populations, but I observed a significant variability also within Mediterranean populations. All birds spent the winter within the Mediterranean basin. Continental Stone-curlews departed earlier in spring and later in autumn and covered longer distances than those from Mediterranean areas. The speed of migration did not change between seasons for continental birds, while Mediterranean individuals migrated faster in spring. The likelihood of departure for autumn migration of GPS-tagged birds increased when temperatures were near or below 0 °C suggesting that Stone-curlews tend to delay departure until weather conditions worsen abruptly. As a consequence of global warming in the Mediterranean, I suggest that the frequency of migratory birds in the considered populations might decrease in the near future. This could affect the distribution of species throughout the year and should be taken into account when targeting conservation measures. In chapter 3 I compared the roosting and foraging behaviour during the pre-migratory and the wintering periods (years 2013-2021) of 28 GPS-tagged Stone-curlews belonging to two populations (Piave and Maremma) living in rather different ecological contexts due to different predominant agricultural practices. In Maremma, Stone-curlews tended to change roost site more frequently than in the Piave area, at least in winter, perhaps because of the greater hunting disturbance. The wide distribution of roost sites in Maremma suggested that there was a high availability of suitable sites in the area, which could at least partially compensate for the effect of disturbance. In both areas the Stone-curlews foraged near the roost site early in the season and then sought feeding areas further afield, perhaps in response to the depletion of resources near the roost sites. The distances travelled in winter were greater than those recorded in the pre-migratory period in both areas. The size of individual foraging areas tended to be larger in the Piave area in both periods, suggesting that food availability was lower in the former possibly due to the dominance of the intensive agricultural practices. This is further suggested by the low degree of overlap between foraging areas used on consecutive nights record in Piave. In conclusion, the collected data indicated that during the non-breeding period the Stone-curlew shows a noticeable degree of ecological and behavioural flexibility that allows the species to exploit different habitat types, including those with a significant degree of intensification of agricultural practices. However, the presence of natural habitats and/or extensive cultivation seems essential to buffer the effect of intensive farming practices and ensure the conservation of the Stone-curlew and possibly of other farmland species. Finally, in chapter 4 I investigated the environmental factors affecting the selection of winter roost and foraging sites of the Stone-curlew in the Grosseto province (Central Italy). To this aim, annual surveys of 24-36 potential roost sites, carried out in the period 2013-2020, were integrated with tracking data collected on eight birds equipped with GPS tags in the period 2013-2017. Roost occurrence probability turned out to be higher in flat terrain but was negatively affected by olive groves cover and road density, a proxy of human disturbance. Other land cover types did not exert a significant effect on roost occurrence probability, but, together with vegetation height, significantly affected roost size. In particular, the highest number of roosting individuals was found in fallows with short vegetation heights (< 5 cm). Contrary to what was observed for roost sites, olive groves were significantly selected by birds during foraging. The use of livestock to control the vegetation cover in traditional olive groves seems to be a disturbing factor during the day, but their grazing activity and the attractive effect of their droppings for invertebrates could favor the foraging activity of the Stone-curlew. In addition to olive groves, birds used a variety of agricultural habitats and showed considerable inter-individual variability in their preferences. These specializations for particular habitats could be an important factor for winter survival, as could help to optimize foraging movements and to reduce competition. The mosaic of different habitats recorded in the province of Grosseto, where extensive agricultural practices are still relatively widespread, could be a critical factor for wintering Stone-curlews. Preserving this heterogeneity is probably fundamental for the conservation of this species in the area. In conclusion, this dissertation focused on the factors affecting the spatial behaviour of the Eurasian Stone-curlew, Burhinus oedicnemus, that could be relevant to predict the future distribution of the species that live in a context as the Western Palearctic, area characterized by a strong effect of climate change and land-use modification

Analisi funzionale degli effetti della suberoanilide dell'acido idrossamico (SAHA) sulla riprogrammazione di fibroblasti in fotorecettori.

Difetti genetici di alcune proteine localizzate in specifici fotorecettori possono portare alla degenerazione di quest’ultimi e possono essere la causa di alcune patologie, tra cui la retinite pigmentosa, che può portare alla progressiva perdita della vista. Attualmente non esistono farmaci che prevengono o curano la degenerazione retinale. Approcci rigenerativi possono fornire una cura per la cecità andando a garantire un’adeguata fonte di cellule transfettabili. La generazione di precursori di fotorecettori ottenuta tramite riprogrammazione genetica di cellule somatiche ha molti vantaggi rispetto all’utilizzo di cellule staminali embrionali (ES) e cellule staminali pluripotenti indotte (iPS) in termini di sicurezza e di numero di cellule. In questa tesi abbiamo testato l’effetto della suberoylanilide dell’acido idrossamico (SAHA), un inibitore delle deacetilasi degli istoni (HDAC) di classe I e II, sulle proprietà funzionali di precursori di fotorecettori a bastoncello generati in vitro da fibroblasti embrionali di topo (MEFs). I nostri risultati indicano che dopo il trattamento di SAHA, le cellule precursori dei bastoncelli esprimono i canali del cGMP, che sono regolati dal cGMP endogeno. Gli effetti del SAHA sono specifici per i precursori dei bastoncelli e richiedono un tempo di induzione di alcuni giorni. Tuttavia il trattamento con SAHA non è riuscito a promuovere l’espressione dell’attività fosfodiesterasica. I nostri dati suggeriscono che lo stato epigenetico dei fibroblasti di topo non permette l’acquisizione delle caratteristiche dei bastoncelli, e le modificazioni posttraduzionali in aggiunta all’acetilazione degli istoni possono ostacolare la trascrizione di geni di specifici fotorecettori nei fibroblasti riprogrammati

L'impiego dei Dispositivi Medici nella ventilazione dei pazienti Covid in area critica: esperienza dell'Ospedale di Livorno.

Nel dicembre 2019 in Cina un nuovo virus, denominato SARS-CoV-2, ha causato una serie di casi di polmonite associati ad elevata mortalità. Il virus si è rapidamente diffuso in tutto il mondo, tanto che l’11 marzo 2020 la WHO ha dichiarato lo stato di pandemia. L’infezione da SARS-CoV-2 è in grado di determinare una sindrome multiorgano, con principale coinvolgimento polmonare e insufficienza respiratoria acuta: si arriva ad un blocco totale dell’attività respiratoria, rendendo indispensabile l’assistenza ventilatoria. In seguito all’avvento della pandemia sono stati messi a punto piani di riorganizzazione delle reti ospedaliere che hanno previsto il potenziamento dei reparti di Terapia Intensiva e sub-intensiva con aumento di posti letto, dotazioni strumentali e aumento massiccio dei dispositivi di supporto respiratorio non invasivo o di ventilazione meccanica. La fornitura dei ventilatori meccanici ha costituito un problema nelle fasi iniziali, in considerazione dell’elevato numero contemporaneo di pazienti critici ricoverati in Terapia intensiva. In questo contesto la Farmacia Ospedaliera ha assunto un ruolo fondamentale nello stoccaggio, gestione e distribuzione di questi dispositivi e ha collaborato strettamente con i clinici dei reparti Covid e Terapia intensiva. Lo scopo di questa tesi è quella di condurre un’analisi delle caratteristiche dei vari dispositivi necessari per il supporto respiratorio confrontandoli e definendo chiaramente le varie indicazioni d’uso. Inoltre è stata condotta un’analisi sull’utilizzo di questi dispositivi in vari periodi dell’emergenza Covid al fine di valutare e verificare se le maggiori conoscenze acquisite sul virus e sul decorso della malattia hanno portato anche ad un cambiamento dei protocolli clinici e quindi di utilizzo di questi dispositivi. Tutto ciò anche in relazione alla definizione di nuove linee guida nazionali ed internazionali. Sono stati pertanto analizzati i pazienti Covid afferenti al Reparto di Terapia Intensiva dell’Ospedale di Livorno andando a valutare le caratteristiche cliniche, il decorso della malattia, il tasso di mortalità ed il tipo di supporto respiratorio utilizzato. Infine è stato valutato l’impatto economico sulla spesa farmaceutica che ne è derivato dall’utilizzo massiccio di questa tipologia di dispositivi rispetto al periodo pre-Covid

NON PEPTIDIC 14-3-3 INHIBITORS AND THE USE THEREOF

The present invention refers to compounds with inhibitory activity against 14-3-3 proteins and their use in the treatment of tumor, in particular chronic myeloid leukemia. The invention also provides methods for the identification of14-3-3 protein inhibitors

Comportamento spaziale dell’occhione in aree caratterizzate da diversi livelli di antropizzazione

Lo studio del comportamento spaziale di una specie in differenti contesti ambientali è fondamentale per comprenderne l’ecologia e per predire la sua risposta a varie tipologie di impatto antropico. L’occhione (Burhinus oedicnemus) è una specie di interesse conservazionistico in Europa, ma nonostante questo, gli studi sulla sua ecologia sono ancora in numero ridotto e raramente basati su dati di tracking individuale. In questo è stato analizzato il comportamento spaziale in periodo riproduttivo e non riproduttivo di animali marcati con GPS data-logger nel periodo 2013-2019 in due popolazioni italiane (provincia di Grosseto e provincia di Treviso) che nidificano in aree caratterizzate da diversi livelli di antropizzazione. La popolazione grossetana insiste su un’area dominata dal pascolo ovino, mentre quella trevigiana nidifica prevalentemente in ambiente fluviale entro un’area dominata da colture intensive. È stata valutata la fedeltà degli animali alle aree frequentate durante le ore diurne (territori di nidificazione/roost post-riproduttivi) e a quelle trofiche frequentate durante la notte. I risultati hanno mostrato una significativa plasticità dei movimenti degli animali prevalentemente legata al relativo dinamismo dell’ambiente di nidificazione. Questa capacità di rispondere ai cambiamenti ambientali spesso di origine antropica rappresenta un fattore importante per le prospettive di conservazione della specie nel medio/lungo periodo

Genetic and metabolic labelling of Extracellular Vescicles: a new tool for biogenesis studies

It has been described that the Human Immunodeficiency Virus (HIV)-1 Nef protein associates with exosomes through anchoring its N-terminal myristoylation to lipid raft microdomains at the endosome membranes. To study Nef association with exosomes, we used a Nef protein mutant defective for all anti-cellular Nef functions that presents a N-terminal palmitoylation, fused at C-terminal with Green Fluorescent Protein (GFP). To demonstrate that Nefmut-GFP goes to compartments like late endosomes or other organelles involved in exosomes biogenesis, we used a novel methodology developed in our laboratory to metabolically label exosomes by incubating cells with a red fluorescent fatty acid BODIPY 558/568 C12 (C12). The lipid is readily taken up by cells and transformed into phospholipids that will ultimately form the exosome lipid bilayer. By transfecting HEK293 cells with the Nefmut-GFP vector and pulsing them with C12 we could purify exosomes containing Nefmut-GFP (Nef-GFP exo) and/or C12 (C12 exo). Fluorescent exosomes were characterized for typical exosomes markers and were analyzed by density gradients showing that Nef-GFP exo could be separated in two distinct peaks whereas C12 exo displayed only one fluorescent peak. Furthermore, they could be sorted by Fluorescence-activated Cell Sorting (FACS) that can be further characterized. In conclusion Nefmut-GFP efficiently incorporates into exosomes that display typical exosome markers (CD63, CD81, CD9, Alix, TSG101). NanoSight analysis showed that vesicles have a mean diameter of 124 nm, compatible with the dimensions of the exosomes reported in literature. Analysis pre-sorting by FACS of double labelled Nefmut-GFP/C12 exosomes showed that the majority of exosomes had incorporated the Nefmut-GFP protein, (92,4% ), of which 40% of the total labeled exosomes is double labeled with Nefmut-GFP and C12 fluorescent lipid. Density gradient separation of double labelled Nefmut-GFP/C12 exosomes showed two distinct peaks for Nefmut-GFP exo whereas C12 exo displayed only one fluorescent peak. Distribution analysis of exo markers by western blot suggested we may have unraveled two different populations of exosomes that can be further characterized

Metabolically labeled exosomes for biogenesis and functional studies

Introduction: Exosomes are small Extracellular Vesicles (sEV) formed by an endosomal route by inward budding of the lateendosome/multivesicular body (MVB) membrane. Despite in recent years much progress has been made to better define sEVcomposition and biogenesis pathways, their small size and heterogeneity pose challenges to find new reliable labelling strategies toidentify specific exosome populations. We developed an innovative methodology to metabolically label fluorescent sEV throughthe use of a fluorescent lipid (BODIPY C16) that is readily internalized by cells and is transformed into phospholipids which willform part of the lipid bilayer of the secreted vesicles.Methods: Fluorescent sEV secreted in the conditioned media of melanoma cells pulsed with BODIPY FL C16 were purifiedby differential ultracentrifugation, quantified by Flow Cytometry (FC) and Nanoparticle Tracking Analysis (NTA), sorted byFluorescence Activated Cell Sorting (FACS) and further characterized by density gradient separation and Western Blot analysisfor typical sEVs markers. Colocalization studies were performed by confocal microscopy and electron microscopy.Results: Confocal images showed colocalization of BODIPY lipids with lipid transformation sites such as ER and mitochondriaand with specific markers of late endosomes/MVB or other organelles (tetraspanins, Golgi markers, lysosomes) but not with the of ABSTRACTplasma membrane. Secretion of fluorescent sEV (Bodipy sEV) was followed over time showing an early release of Bodipy sEVinto the extracellular medium with a constant ratio of Bodipy sEV/total EVs, as determined by NTA, up to 6 hours. Bodipy sEVsecreted in the conditioned media purified by differential ultracentrifugation were separated by density gradient fractionation.Fractions analysed by FC displayed a single low density peak at 1,08-1,09 g/ml that is detergent sensitive demonstrating thatfluorescent particles are indeed lipid vesicles and contain tetraspanins (CD63, CD81 and CD9), syntenin and ESCRT componentswhen analysed by Western Blot. Electron microscopy analysis of ultracentrifuged and sorted Bodipy sEV showed that BodipysEVhavethetypicalshapeandsize(about80nm)ofasubpopulationofsEVoftenreferredtoassmallexosomes(Exo-S).Finally,colocalization studies of single Bodipy sEV with tetraspanins fluorescent antibodies showed colocalization of Bodipy sEV withCD63, CD81 and CD9.Summary/Conclusion: Taken together these results show a very specific and effective labelling of a discrete sEV subpopulationthat can be further exploited for biogenesis, internalization and functional studies. This work was supported by the Italian Ministry of Health (grant RF-2019-12369719