Artrite reumatoide e analisi proteomica della saliva

L’Artrite Reumatoide (AR) è una malattia autoimmune sistemica ad eziologia sconosciuta, caratterizzata dallo sviluppo di una sinovite cronica responsabile di un danno cartilagineo e di erosioni ossee poliarticolari. L’infiammazione prende avvio da un’attivazione del sistema immunitario contro antigeni self e numerose sono le molecole che sono state proposte quali possibili auto antigeni: il collagene di tipo II, la glicoproteina-39 dei condrociti umani, la glucosio-6-fosfato isomerasi e alcune HSPs secrete durante stress, come la chaperonina BiP (GRP78). Recentemente, per l’identificazione di specifici biomarkers di malattia, sono stati avviati diversi studi di proteomica sul siero, sul liquido sinoviale e sulle cellule (sinoviociti e linfociti) di pazienti con diagnosi di AR. Scopo del lavoro – L’ipotesi di lavoro che ha guidato questo studio è stata quella di identificare, attraverso l’utilizzo dell’elettroforesi bidimensionale (2DE) combinata con la spettrografia di massa (MS), le proteine contenute nella saliva di pazienti affetti da AR e di confrontare tale pattern proteico salivare con quello di soggetti sani, al fine di evidenziare le eventuali alterazioni dell’espressione proteica che si manifestano nella malattia. Pazienti e metodi – Nello studio sono stati arruolati 20 pazienti con diagnosi di AR formulata in base ai criteri dell’American College of Rheumatology (ACR) e 20 soggetti sani confrontabili per sesso e per età a costituire un gruppo di controllo. La caratterizzazione del profilo proteico salivare di ciascun campione è stata effettuata con l’elettroforesi bidimensionale e gli spot proteici di interesse sono stati identificati con la spettrometria di massa (MALDI-TOF mass spectrometry). Risultati - L’analisi delle immagini ha permesso di identificare 10 spots proteici significativamente up-regolati nei pazienti con AR rispetto al gruppo di controllo. Tali spots sono confluiti in 8 diverse proteine: calgranulina A, calgranulina B, proteina epidermica legante gli acidi grassi (E-FABP), 6-fosfogluconato deidrogenasi (6-PGDH), perossiredossina 5 (PRX5), Apolipoproteina A-1, chaperonina GRP78/BiP e proteine della famiglia 14-3-3. L’analisi Western blot (WB) è stata utilizzata per verificare i risultati ottenuti dall’analisi dei gels dell’elettroforesi bidimensionale allargando l’indagine, per le proteine di maggiore interesse GRP78/BiP e 14.3.3, ad un gruppo di controllo patologico rappresentato da 10 pazienti affetti da artropatia psoriasica. Ciò ha permesso di identificare le due isoforme specifiche delle proteine 14-3-3, ovvero la gamma e la sigma, e di dimostrare un’up-regolazione della proteina GRP78/BiP nei soggetti affetti da AR, sia rispetto ai controlli sani che ai controlli patologici. Conclusioni - Lo studio ha consentito di evidenziare il ruolo potenziale dell’analisi proteomica salivare nella ricerca di biomarkers dell’AR. In particolare i risultati ottenuti suggeriscono la possibilità di identificare diversi aspetti della malattia attraverso l’analisi delle proteine contenute nei campioni di saliva. Da questo punto di vista, le proteine 14.3.3 potrebbero rappresentare indicatori non specifici di danno cartilagineo, mentre la proteina GRP78/BiP potrebbe esser considerata come un vero e proprio biomarker dell’AR

Development of nature inspired antiplasmodial hits possessing the thiazinoquinone pharmacophore

Malaria accounts globally for more than 200 million new cases and 438,000 deaths per year. Since malaria is a disease of worldwide implications, combating it is one of the highest priority programs of the WHO. A worrisome increase in the number of fatal cases has been registered in recent years and it is principally due to the diffusion of multi-drug resistant strains of Plasmodium, making less effective the limited armamentarium of available drugs. Therefore, there is an urgent need of new antimalarial drugs with high efficacy against resistant strains and broad stage mode of action. To reach these challenging aims, the identification and selection of new lead compounds constitutes a crucial point. In this regard, nature remains an ever evolving resource. Recently, the antiplasmodial activity of marine secondary metabolites characterized by a quinone scaffold has been reported. In particular, it is worth to point out that a number of quinones have been shown to be effective antimalarials. The observed effects are most likely related to the most prominent chemical feature of these kind of molecules, that is their ability to undergo redox reaction i) shuttling electrons from reduced flavoproteins to acceptors such as hemoglobin-associated or free Fe(III)-protoporphyrin IX or ii) inhibiting the mitochondrial electron transport chain. In this context, recently, we were inspired by two marine metabolites Aplidinone A and B isolated from the Mediterranean ascidian Aplidium conicum, and we developed a series of synthetic analogues featuring the thiazinoquinone chemotype present in the natural metabolites with simplified side chains and different substituents. Manipulation of this chemical scaffold afforded additional analogues with improved pharmacological proprieties compared to the starting hits identified in the previous series

Investigating the antiparasitic potential of the marine sesquiterpene avarone, its reduced form avarol, and the novel semisynthetic thiazinoquinone analogue thiazoavarone

The chemical analysis of the sponge Dysidea avara afforded the known sesquiterpene quinone avarone, along with its reduced form avarol. To further explore the role of the thiazinoquinone scaffold as an antiplasmodial, antileishmanial and antischistosomal agent, we converted the quinone avarone into the thiazinoquinone derivative thiazoavarone. The semisynthetic compound, as well as the natural metabolites avarone and avarol, were pharmacologically investigated in order to assess their antiparasitic properties against sexual and asexual stages of Plasmodium falciparum, larval and adult developmental stages of Schistosomamansoni (eggs included), and also against promastigotes and amastigotes of Leishmania infantum and Leishmania tropica. Furthermore, in depth computational studies including density functional theory (DFT) calculations were performed. A toxic semiquinone radical species which can be produced starting both from quinone- and hydroquinone-based compounds could mediate the anti-parasitic effects of the tested compounds

Indagine di mercato su Realtà Aumentata e Virtuale

Questo rapporto costituisce il risultato della ricerca su possibili aree di interesse che possano attirare l’attenzione di utenti terzi (rispetto al partenariato del progetto 3DLab-Sicilia) al fine di individuare scenari di utilizzo e valorizzazione degli asset (VR Cave, tecnologie, strumenti, competenze, ecc.) sviluppati nel progetto e messi a disposizione dai partner. L’obiettivo della ricerca è quello di individuare aree di applicazione delle tecnologie di Realtà Aumentata, Mista e Virtuale (AR/MR/VR), soprattutto quelle non esplorate già dal progetto 3DLab-Sicilia, che possano essere opportunamente comunicate e diffuse, per raccogliere la manifestazione di interesse, da parte di piccole e medie imprese del territorio, a partecipare al “contest” che verrà attivato nel corso della seconda parte del progetto finanziato. La ricerca è stata effettuata da Xenia Progetti, consulente del Parco Scientifico e Tecnologico della Sicilia per la fornitura di servizi relativi alla realizzazione del WP4 del progetto 3DLab-Sicilia. Un ulteriore scopo dell’indagine è quello di fornire uno strumento al partenariato 3DLab-Sicilia per rendere sostenibile l’iniziativa, dopo la conclusione del progetto, attraverso delle indicazioni su aree tematiche e possibili applicazioni verso le quali dirigere l’azione di promozione dei risultati del progetto

Salivary gland ultrasonography: a highly specific tool for the early diagnosis of primary Sjögren's syndrome

INTRODUCTION:Recently, a great interest has arisen for salivary gland ultrasonography (SGUS) as a valuable tool for the assessment of major salivary gland involvement in primary Sjögren's syndrome (pSS. The aims of this study were to test the accuracy of SGUS for the early detection of pSSand to compare the diagnostic performance of SGUS with minor salivary gland biopsy (MSGB) and unstimulated salivary flow (USFR) in this context. METHOD:Patients with suspected pSS and symptoms duration of ≤5 years were consecutively enrolled in this study. The diagnosis of pSS was made according to the AECG criteria. SGUS was performed by two radiologists blinded to the diagnosis and a previously reported ultrasound scoring system (De Vita et al. 1992, cut-off ≥ 1) was used to grade the echostructure alterations of the salivary glands. Statistical analysis was performed using SPSS v16. RESULTS: This study included 50 pSS patients and 57 controls with no-SS sicca symptoms. The mean(SD) age of the pSS group was lower than non-SS group (47(13) vs 53(12)yrs, p = 0.006). No further differences between the two groups were observed. Patients with pSS showed a significantly higher SGUS score in comparison with controls (mean(SD) = 2.1(1.8) vs 0.0(0.4), p = 0.000). The SGUS cut-off ≥ 1 showed a sensitivity (SE) of 66 %, a specificity (SP) of 98 %, a positive predictive value (PPV) of 97 % and a negative predictive value (NPV) of 73 % for pSS diagnosis. The SGUS score correlated also with patients' MSGB/FS and USFR. CONCLUSIONS: This study confirmed the good performance of SGUS for the early non-invasive diagnosis of pSS. Further research in larger international cohort of patients is mandatory in order to assess the role of SGUS in the diagnostic algorithm of pSS

MS Dereplication for Rapid Discovery of Structurally New or Novel Natural Products

In order to accelerate the isolation and characterisation of structurally new or novel natural products, it is crucial to develop efficient strategies that prioritise samples with greatest promise early in the workflow so that resources can be utilised in a more efficient and cost-effective manner. Two complementary approaches have been developed: One is based on targeted identification of known compounds held in a database based on high resolution MS and predicted LC retention time data [1]. The second is an MS metrics-based approach where the software algorithm calculates metrics for sample novelty, complexity, and diversity after interrogating databases of known compounds, and contaminants. These metrics are then used to prioritise samples for isolation and structure elucidation work [2]. Both dereplication approaches have been validated using natural product extracts resulting in the isolation and characterization of new or novel natural products

Direct event location techniques based on Adams multistep methods for discontinuous ODEs

In this paper we consider numerical techniques to locate the event points of the differential system x′=f(x), where f is a discontinuous vector field along an event surface splitting the state space into two different regions R1 and R2 and f(x)=fi(x) when x∈Ri, for i=1,2 while f1(x)≠f2(x) when x∈Σ. Methods based on Adams multistep schemes which approach the event surface Σ from one side only and in a finite number of steps are proposed. Particularly, these techniques do not require the evaluation of the vector field f1 (respectively, f2) in the region R2 (respectively R1) and are based on the computation–at each step– of a new time ste

Upadacitinib effectiveness and factors associated with minimal disease activity achievement in patients with psoriatic arthritis: preliminary data of a real-life multicenter study

Background Upadacitinib (UPA) is a selective JAK inhibitor recently approved for the treatment of psoriatic arthritis (PsA). In this post-approval study, we aimed to evaluate the effectiveness and safety of UPA over 24 weeks and identify clinical predictors of response, in a multicentric cohort of patients affected by PsA.Methods One hundred and twenty-six patients with PsA treated with UPA were enrolled in 10 Italian centres. UPA effectiveness outcomes, such as the proportion of patients with MDA status, DAPSA remission, and low disease activity, ASDAS-CRP inactive and low disease activity, and change from baseline in DAPSA and ASDAS-CRP scores, were evaluated every 12 weeks until week 24. The proportion of DAPSA minor, moderate, and major improvement, and ASDAS clinically important improvement (CII) and major improvement (MI) were considered as well. All treatment-related adverse events were collected during the observation period. Clinical predictors of MDA response at week 24 were evaluated through multivariate analysis.Results At baseline, 124/126 (98%) and 54/126 (43%) patients showed peripheral and axial involvement, respectively; 110 (87%) patients were intolerant or resistant to biologic DMARDs.At 24 weeks, MDA status, DAPSA remission, and ASDAS-CRP inactive disease were achieved in 47%, 23%, and 48% of patients, respectively. Minor, moderate, and major DAPSA improvement was observed in 67%, 39%, and 23%, respectively; while 65% and 35% achieved ASDAS-CRP CII and MI, respectively. The mean change from baseline was 15.9 +/- 13.5 (p < 0.001) for DAPSA and 1.21 +/- 0.97 (p < 0.001) for ASDAS-CRP. thirteen patients (10%) discontinued UPA due to a lack of efficacy or non-serious adverse events. No serious adverse events were observed. Male gender (OR 2.54, 95% CI 1.03-6.25 p = 0.043), being naive to biological DMARDs (OR 4.13, 95% CI 1.34-12.71, p = 0.013) and elevated baseline CRP (OR 2.49, 95% CI 1.02-6.12, p = 0.046) were associated with MDA response at week 24.conclusions this is one of the first real-life studies supporting the effectiveness of UPA and its safety profile in PsA patients. Furthermore, the study identifies predictors of MDA response to UPA treatment at 6 months

Dose-Dependent Impairment of the Immune Response to the Moderna-1273 mRNA Vaccine by Mycophenolate Mofetil in Patients with Rheumatic and Autoimmune Liver Diseases

The purpose of this study was to evaluate the efficacy and safety of the Moderna-1273 mRNA vaccine for SARS-CoV-2 in patients with immune-mediated diseases under different treatments. Anti-trimeric spike protein antibodies were tested in 287 patients with rheumatic or autoimmune diseases (10% receiving mycophenolate mofetil, 15% low-dose glucocorticoids, 21% methotrexate, and 58% biologic/targeted synthetic drugs) at baseline and in 219 (76%) 4 weeks after the second Moderna-1273 mRNA vaccine dose. Family members or caretakers were enrolled as the controls. The neutralizing serum activity against SARS-CoV-2-G614, alpha, and beta variants in vitro and the cytotoxic T cell response to SARS-CoV-2 peptides were determined in a subgroup of patients and controls. Anti-SARS-CoV-2 antibody development, i.e., seroconversion, was observed in 69% of the mycophenolate-treated patients compared to 100% of both the patients taking other treatments and the controls (p < 0.0001). A dose-dependent impairment of the humoral response was observed in the mycophenolate-treated patients. A daily dose of >1 g at vaccination was a significant risk factor for non-seroconversion (ROC AUC 0.89, 95% CI 0.80-98, p < 0.0001). Moreover, in the seroconverted patients, a daily dose of >1 g of mycophenolate was associated with significantly lower anti-SARS-CoV-2 antibody titers, showing slightly reduced neutralizing serum activity but a comparable cytotoxic response compared to other immunosuppressants. In non-seroconverted patients treated with mycophenolate at a daily dose of >1 g, the cytotoxic activity elicited by viral peptides was also impaired. Mycophenolate treatment affects the Moderna-1273 mRNA vaccine immunogenicity in a dose-dependent manner, independent of rheumatological disease

LA PALAFITTA DI TOMBOLA DI CEREA (VR). LO SCAVO 1999.

La palafitta \ue8 stata scoperta nel 1955 all'interno della Valle del Menago, poche centinaia di metri a Sud del gruppo di case di Tombola di Cerea (VR). Dopo un primo scavo effettuato nello stesso anno da parte di Francesco Zorzi, direttore del Museo Civico di Storia Naturale di Verona, le ricerche sono riprese nel 1999 da parte della Soprintendenza Archeologica del Veneto. I risultati di queste ricerche hanno accertato che l'abitato preistorico era posto su palafitta all'interno di un ambiente di tipo palustre. Sono conservati solo dei pali verticali e alcune travi orizzontali, che fanno ipotizzare l'esistenza di una piattaforma lignea. Gli elementi strutturali della palafitta trovano confronti in altri abitati d'ambiente umido della media et\ue0 del Bronzo. L\u2019inquadramento geomorfologico finalizzato alla ricostruzione paleogeografica mostra che il sito si colloca al centro della Paleovalle del f. Menago, un\u2019incisione larga alcune centinaia di metri e profonda oltre una decina di metri, alla base di un potente spessore di torbe. Gli studi pregressi, relativi all\u2019origine della profonda incisione generata dall\u2019omonimo fiume di risorgiva, si richiamano principalmente a fattori geodinamici legati a linee tettoniche sepolte, quali moventi principali per le incisioni causate dalle diversione di canali di rotta atesini richiamati verso il bacino delle Valli Grandi Veronesi, verificatesi in et\ue0 tardoglaciale-olocenica antica. L\u2019analisi di nuove litostratigrafie rilevate su esposizioni individuate all\u2019interno della paleovalle, ha condotto a riscontrare la presenza di uno spesso e pressoch\ue9 continuo riempimento di sedimenti organico-torbosi, che contrassegnano gli accrescimenti e le sedimentazioni verificatesi a partire dai livelli basali della stazione palafitticola, deposizioni che sono perdurate sino a date piuttosto recenti. Una serie di dettagliati confronti operati con i termini delle sequenze di riempimento torboso-detritiche del bacino che si sviluppava all\u2019esterno delle arginature del vicino e coevo sito di Fondo Paviani, ha portato a verificare un\u2019analoga evoluzione negli accrescimenti che contrassegnano i tratti superiori di queste due importanti successioni alluvio-organiche. Si giunge in definitiva a far intervenire una medesima incidenza di fattori antropici e climatici quali concause principali nello sviluppo dei riempimenti sommitali della Paleovalle del Menago, in corrispondenza della stazione palafitticola della Tombola di Cerea e del sito arginato di Fondo Paviani. Lo scavo ha restituito un notevole quantitativo di materiale ceramico (ca. 620 kg) e una percentuale minoritaria di manufatti in bronzo, terracotta e osso-corno. Il presente contributo si concentra sull'analisi crono-tipologica dei materiali focalizzando l'attenzione sulla ceramica di impasto medio-fine ai fini dell'inquadramento complessivo e del confronto con materiale proveniente da siti geograficamente limitrofi. Le datazioni assolute delle strutture lignee, ottenute dall\u2019applicazione della dendrocronologia e dal radiocarbonio, collocano gli elementi verticali di Tombola di Cerea nella seconda met\ue0 del XV secolo cal BC, o al pi\uf9 tardi nei primi decenni del XIV secolo cal BC; essi sono stati ottenuti da alberi di quercia abbattuti in momenti successivi, a intervalli di circa 10 anni. Le ricerche archeobotaniche hanno compreso un\u2019analisi palinologica condotta su 18 campioni presi da una carota prelevata all'interno dell\u2019abitato palafitticolo, ed un\u2019analisi carpologica e xilo-antracologica su due campioni provenienti da unit\ue0 stratigrafiche cronologicamente corrispondenti alla parte basale della carota. L\u2019analisi dei resti vegetali ha permesso di disegnare il quadro ambientale dell\u2019insediamento. Il luogo era ricco di ambienti umidi soprattutto ripariali, con boscaglie d\u2019igrofile e con elofite, con situazioni di prati periodicamente inondati e fossati poco profondi. La presenza antropica \ue8 suggerita dalla coltivazioni di cereali e altri indicatori antropogenici. Inoltre i dati carpologici e/o pollinici testimoniano la disponibilit\ue0 nell\u2019area di varie piante con frutti eduli, come prugnolo, nocciolo, ribes, noce e vite. Il ritrovamento di reperti di Baldellia ranuncoloides -una specie oggi minacciata sottolinea come le ricerche archeobotaniche forniscano informazioni anche dal punto di vista naturalistico utili per conoscere meglio la storia di specie oggi in rarefazione. L\u2019insieme faunistico \ue8 composto complessivamente da 1995 reperti ossei. Quasi l\u201980% dei resti \ue8 stato identificato a livello specifico. La maggior parte dei resti appartiene alle specie domestiche: bue, pecora, capra e maiale, seguite da cavallo e cane. La distribuzione delle classi d\u2019et\ue0 mostra come l\u2019allevamento degli Ungulati fosse finalizzato non solo all\u2019ottenimento di carni, ma anche alla produzione di latte e lana. La pratica della caccia, volta allo sfruttamento di specie di ambiente forestale (cervo, capriolo, cinghiale), appare un\u2019attivit\ue0 marginale. Le tracce di macellazione sono numerose e attestano l\u2019utilizzo di strumenti sia litici sia metallici. Durante lo studio sono stati riconosciuti 18 manufatti in materia dura animale, ben conservati, che hanno permesso l\u2019identificazione delle tracce di lavorazione utili per la ricostruzione delle catene operative.The pile dwelling was discovered in 1955 in the valley of the river Menago, just a few hundred meters south of the settlement of Tombola di Cerea (VR). A first excavation was carried out that very year by Francesco Zorzi, director of the Museo Civico di Storia Naturale of Verona; the digging has been resumed in 1999 by the Soprintendenza Archeologica of Veneto. The results of these excavations have ascertained that the prehistoric dwelling site was built in a marshy environment. Only some vertical poles and some horizontal beams were preserved, which leads to the hypothesis that there existed a wooden platform on which the sheds were standing. The structural elements of the palafitte find correspondents in others settlements in moist environment from the Middle Bronze Age. The paleogeographic reconstruction of the immediate hinterland confirm the location of the site at the center of the Menago River Paleovalley, an incision a few hundred meters wide and more than ten meters deep, at the base of a thick layer of peat. Previous studies, related to the origin of the deep incision generated by the homonymous spring-line river, mainly refer to geodynamic factors linked to buried tectonic lines, as main motives for the incisions caused by the diversion of Adige breach-channels captured towards the basin of Great Veronese Valleys, which occurred in the late Lateglacial-Holocene age. The analysis of new lithostratigraphies detected on exposures inside the paleovalley, has led to find the presence of a thick and almost continuous filling of organic-peaty sediments, which mark the accretions and the sedimentations verified starting from the basal levels of the pile-dwelling site, depositions that have lasted until fairly recent dates. A series of detailed comparisons made with the terms of the peat-debris-filling sequences of the basin that developed outside the embankments of the near and coeval Fondo Paviani site, led to verify a similar evolution in the growths that mark the upper reaches of these two important alluvio-organic successions. Ultimately, is claimed that the same incidence, of anthropogenic and climatic factors, is involved as the main contributors to the development of the upper fills of the Menago Paleovalley, in correspondence with the pile-dwelling station of the Tombola di Cerea and the embanked site of Fondo Paviani. A great number of ceramic fragments (ca 620 kg) and a smaller amount of bronze, terracotta and bone-horn artefacts had been found. This paper focus on chrono-typological analysis of the archaeological finds, in particular on ceramic of the medium-fine tipe. The goal is the overall classification artefacts, accompanied by a comparison made from materials found in geographically nearbysites. Dendrochronology and radiocarbon allow to date the wooden structures to the second half of the 15th Century cal BC, or in the first decades of the 14th Century cal BC at the latest; the vertical posts come from oak tree felled down once about every ten years. Archaeobotanical study includes pollen analysis carried out on 18 subsamples taken from a core collected in the pile-dwelling settlement and carpological and xilo-anthacological analysis, from 2 samples taken from layers (stratigraphic units) corresponding to the bottom of the core. The results allows to draw the environmental framework of the settlement. The pollen diagram shows that the landscape was rich of wet environments: ponds, wet grassland and hygrophilous wood. The human activity was suggested by a number of anthropogenic indicators: cereal fields, ruderals and weeds. Seeds/fruits and pollen data testify to the presence in the area of edible fruits from plants such as blackthorn, hazelnut, currant, walnut and grapevine. The finds of Baldellia ranuncoloides \u2013 a threatened plants today - underline that the archaeobotanical research provide naturalistic information that allow us to better understand the history of plant species becoming rare today. The faunal assemblage is composed by 1995 bone remains. More than 80% of the faunal remains was identified at species level. Domestic animals are the most frequent. Age at death highlights different breeding strategies focused on meat consumption and secondary products. Hunting was barely practiced on wild ungulates (red deer, roe deer, wild boar). Several anthropic marks were identified on the faunal remains emphasizing the use of lithic and metal tools. During the analysis 18 animal hard material artefacts were identified. The good state of preservation allowed us to identify working traces linked to the reduction sequences of the tools