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

Further evidence supporting the role of GTDC1 in glycine metabolism and neurodevelopmental disorders

Copy number variants (CNVs) represent the genetic cause of about 15-20% of neurodevelopmental disorders (NDDs). We identified a similar to 67 kb de novo intragenic deletion on chromosome 2q22.3 in a female individual showing a developmental encephalopathy characterised by epilepsy, severe intellectual disability, speech delay, microcephaly, and thin corpus callosum with facial dysmorphisms. The microdeletion involved exons 5-6 of GTDC1, encoding a putative glycosyltransferase, whose expression is particularly enriched in the nervous system. In a previous study, a balanced de novo translocation encompassing GTDC1 was reported in a male child with global developmental delay and delayed speech and language development. Based on these premises, we explored the transcriptomic profile of our proband to evaluate the functional consequences of the novel GTDC1 de novo intragenic deletion in relation to the observed neurodevelopmental phenotype. RNA-seq on the proband's lymphoblastoid cell line (LCL) showed expression changes of glycine/serine and cytokine/chemokine signalling pathways, which are related to neurodevelopment and epileptogenesis. Subsequent analysis by ELISA (enzyme-linked immunosorbent assay) and HPLC (high-performance liquid chromatography) revealed increased levels of glycine in the proband's LCL and serum compared to matched controls. Given that an increased level of glycine has been observed in the plasma samples of individuals with Rett syndrome, a condition sharing epilepsy, microcephaly, and intellectual disability with our proband, we proposed that the GTDC1 downregulation is implicated in neurodevelopmental impairment by altering glycine metabolism. Furthermore, our findings expanded the phenotypic spectrum of the novel GTDC1-related condition, including microcephaly and epilepsy among relevant clinical features

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