Anti-fibroblast antibodies detected by cell-based ELISA in systemic sclerosis enhance the collagenolytic activity and matrix metalloproteinase-1 production in dermal fibroblasts

Objectives. Antibodies binding to the surface of fibroblasts (anti-fibroblast antibodies: AFA) have been described in systemic sclerosis (SSc). We aimed to assess the effect of AFA on extracellular matrix (ECM) turnover and whether AFA were associated with anti-topoisomerase-I antibody. Methods. IgG were purified from AFA-positive and AFA-negative sera selected within 20 SSc and 20 healthy individuals, and tested on normal dermal fibroblasts, at protein and mRNA level, for their capacity to induce collagen deposition or degradation. Results. Fibroblasts stimulated with AFA-positive but not with AFA-negative and control IgG showed an increased capacity to digest collagen matrix and produce metalloproteinase-1 (MMP-1) while their production of total collagen, type I collagen and tissue inhibitor of metalloproteinase-1 (TIMP-1) was unaffected. The steady-state mRNA levels of MMP-1, COL1A1 and TIMP-1 paralleled the protein levels. AFA-positive IgG did not induce Smad 2/3 phosphorylation, indicating that this transforming growth factor-β signalling pathway was not involved. IL-1 and tumour necrosis factor (TNF) neutralization did not reverse the enhanced production of MMP-1, suggesting a direct effect of AFA on fibroblasts. Finally, anti-topoisomerase-I antibodies were present in 11 of 12 AFA-negative IgG, and an anti-topoisomerase-I monoclonal antibody failed to enhance MMP-1 production, thus indicating a lack of correlation between AFA and anti-topoisomerase-I antibody. Conclusions. These results indicate that SSc antibodies binding to fibroblasts enhance matrix degradation and MMP production events that may favour inflammation but do not directly impact on fibrosis developmen

Mir-101-3p downregulation promotes fibrogenesis by facilitating hepatic stellate cell transdifferentiation during insulin resistance

Insulin resistance (IR) and microRNAs (miRNAs), which regulate cell-to-cell communication between hepatocytes and hepatic stellate cells (HSCs), may intertwine in nonalcoholic fatty liver disease (NAFLD) pathogenesis. The aim of this study was to evaluate whether epigenetics and environmental factors interact to promote progressive NAFLD during IR. We examined the miRNA signature in insulin receptor haploinsufficient (InsR+/ 12) and wild-type (wt) HSCs by RNAseq (n = 4 per group). Then, we evaluated their impact in an IR-NASH (nonalcoholic steatohepatitis) model (InsR+/ 12 mice fed standard or methionine choline deficient (MCD) diet, n = 10 per group) and in vitro. InsR+/ 12 HSCs displayed 36 differentially expressed miRNAs (p < 0.05 vs. wt), whose expression was then analyzed in the liver of InsR+/ 12 mice fed an MCD diet. We found that miR-101-3p negatively associated with both InsR+/ 12 genotype and MCD (p < 0.05) and the histological spectrum of liver damage (p < 0.01). miR-101-3p was reduced in InsR+/ 12 hepatocytes and HSCs and even more in InsR+/ 12 cells exposed to insulin (0.33 \ub5M) and fatty acids (0.25 mM), resembling the IR-NASH model. Conversely, insulin induced miR-101-3p expression in wt cells but not in InsR+/ 12 ones (p < 0.05). In conclusion, IR combined with diet-induced liver injury favors miR-101-3p downregulation, which may promote progressive NAFLD through HSC and hepatocyte transdifferentiation and proliferation

Anti-atherogenic modification of serum lipoprotein function in patients with rheumatoid arthritis after tocilizumab treatment, a pilot study

Lipid metabolism derangement contributes to increased cardiovascular risk in Rheumatoid Arthritis (RA). It is still debated whether and how tocilizumab, an interleukin-6 receptor inhibitor used in active RA, impacts cardiovascular risk. We studied the effect of tocilizumab on the regulation of macrophage cholesterol homeostasis, measuring patient serum ability to respectively load (cholesterol loading capacity, CLC) and discharge (cholesterol efflux capacity, CEC) cells with cholesterol. Patients with RA (n = 8) were studied before and after 4 and 12 weeks of tocilizumab treatment. CLC was measured by a fluorimetric assay of intracellular cholesterol content in human macrophages and CEC was measured for the three main pathways, mediated by the transporters Scavenger Receptor class B-type I (SR-BI), ATP binding cassette-G1 (ABCG1) and-A1 (ABCA1) in specific cell models. After 12 weeks of tocilizumab treatment, serum LDL cholesterol levels were increased, while CLC was reduced. HDL cholesterol levels were unchanged, but CEC was significantly ameliorated for the SR-BI and ABCG1 pathways with respect to baseline. Tocilizumab reduces LDL pro-atherogenic potential despite increasing their serum levels and increases HDL protective activity in RA. The data of our pilot study suggest that tocilizumab regulates lipoprotein function in selected patient populations and lay the groundwork for future larger studies

La difesa dai terremoti in Lombardia: stato dell’arte e prospettive

L’INGV (http://www.mi.ingv.it/) svolge in Lombardia ricerche nel campo della mitigazione del rischio sismico, mediante studi mirati al miglioramento delle conoscenze sulla storia sismica, sul modello strutturale legato al regime tettonico in atto e alla definizione del moto del suolo atteso. Accanto alle indagini necessarie alla caratterizzazione sismogenetica e dei possibili effetti dello scuotimento sismico, si pone l’attività di monitoraggio sismico, che la Sezione di Milano-Pavia espleta mediante la rete accelerometrica (RAIS, http://rais.mi.ingv.it/) che consta di 20 postazioni distribuite in prevalenza sul territorio regionale. In un recente lavoro di sintesi - di studi pluriennali su fonti storiche e di revisioni critiche di materiali pubblicati - si è tentato di colmare l’evidente carenza conoscitiva sulle caratteristiche della sismicità storica dell’area lombarda. Tale carenza è particolarmente evidente se si rapportano le informazioni oggi disponibili sui terremoti del passato in quest’area con quelle relative al settore veneto-friulano. La regione analizzata, compresa tra il bacino del fiume Adda e il Lago di Garda, è stata caratterizzata da alcuni terremoti con Mw>5.5 (es., 1117, Veronese; 1222, Brescia; 1901, Salò) e vari eventi con Mw compresa fra 4.8 e 5.5 (es., 1065, Brescia; 1396, Monza; 1642, Bergamo). Per molti degli eventi sismici fino al 1700 le informazioni sono desumibili soltanto da scarse fonti storiche. La determinazione epicentrale e l’attribuzione della magnitudo per tali eventi sono da considerarsi, pertanto, con una certa cautela al fine di definire le caratteristiche sismiche del territorio. I terremoti del 1117 e del 1222, in tale contesto, rappresentano un’eccezione rispetto ai dati generalmente disponibili. E tuttavia vari problemi tuttora aperti rendono assai difficile l’utilizzo della distribuzione del danno attribuibile a questi eventi nella prospettiva di un’affidabile parametrizzazione. Le indagini geologico-strutturali e di geologia del Quaternario, finalizzate a definire un quadro strutturale compatibile con il regime tettonico in atto, sono necessarie per giustificare la storia sismica e, in sostanza, per definire il comportamento sismogenetico della regione. Le geometrie dei sistemi di faglia attivi alpini sono ora sufficientemente noti. Le conoscenze permettono di formulare ipotesi sismotettoniche relative all’origine dei terremoti dell’area gardesana e del Bresciano. In via di definizione sono invece le geometrie dei fronti appenninici, cui sono attribuibili i terremoti al di sopra della soglia nel settore padano. Le ricerche attuali sono altresì indirizzate ad una migliore caratterizzazione del complesso settore compreso tra la parte meridionale del Lago di Garda (area di Sirmione), Verona e Mantova, all’interno del quale potrebbe collocarsi l’area epicentrale del terremoto del 1117 (o una delle aree epicentrali, qualora si considerasse questo evento come rappresentato da una sequenza sismica). Uno dei settori regionali con maggiore frequenza di eventi sismici è l’area gardesana occidentale, per questo motivo la rete di monitoraggio presenta una notevole densità di postazioni nel Bresciano e se ne è pianificato l’addensamento nel Veronese. Nel corso del 2007, la rete ha consentito la registrazione di 516 forme d’onda relative a 28 eventi locali e regionali (di cui una decina localizzati nell’area citata) con magnitudo da 1.3 a 4.2, di cui sono stati calcolati i parametri di interesse ingegneristico. L’analisi delle registrazioni ha permesso di ricavare informazioni utili per il calcolo di scenari di scuotimento. Un esempio di taali applicazioni è rappresentato dallo scenario realizzato utilizzando come terremoto di riferimento l'evento del 24 Novembre 2004 (M 5.2)

The role of insulin resistance in nonalcoholic steatohepatitis and liver disease development : a potential therapeutic target?

Insulin resistance (IR) is defined by the inability of insulin to exert its metabolic actions, due to impaired activation of intracellular insulin signaling. This condition is caused by genetic defects or by environmental conditions, among which the most common is obesity. Systemic IR determines the development of hepatic fat accumulation, which can progress to nonalcoholic steatohepatitis, cirrhosis and hepatocellular carcinoma, and is a major determinant of liver disease independently of coexisting factors. Therefore, insulin-sensitizing drugs are currently under evaluation to improve steatohepatitis. Indeed, manipulation of nuclear hormone receptors is already under scrutiny for liver disease prevention by amelioration of IR, whereas NOTCH signaling inhibition represents a novel approach. Nevertheless, further research is warranted to better understand the mechanism linking IR to progressive fibrogenesis in the absence of inflammation and to identify novel drug targets

Mother-to-child transmission of KPC-producing Klebsiella pneumoniae: Potential relevance of a low microbial urinary load for screening purposes

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Complement activation predicts negative outcomes in COVID-19: The experience from Northen Italian patients

Coronavirus disease 19 (COVID-19) may present as a multi-organ disease with a hyperinflammatory and prothrombotic response (immunothrombosis) in addition to upper and lower airway involvement. Previous data showed that complement activation plays a role in immunothrombosis mainly in severe forms. The study aimed to investigate whether complement involvement is present in the early phases of the disease and can be predictive of a negative outcome. We enrolled 97 symptomatic patients with a positive RT-PCR for SARS-CoV-2 presenting to the emergency room. The patients with mild symptoms/lung involvement at CT-scan were discharged and the remaining were hospitalized. All the patients were evaluated after a 4-week follow-up and classified as mild (n. 54), moderate (n. 17) or severe COVID-19 (n. 26). Blood samples collected before starting any anti-inflammatory/immunosuppressive therapy were assessed for soluble C5b-9 (sC5b-9) and C5a plasma levels by ELISA, and for the following serum mediators by ELLA: IL-1β, IL-6, IL-8, TNFα, IL-4, IL-10, IL-12p70, IFNγ, IFNα, VEGF-A, VEGF-B, GM-CSF, IL-2, IL-17A, VEGFR2, BLyS. Additional routine laboratory parameters were measured (fibrin fragment D-dimer, C-reactive protein, ferritin, white blood cells, neutrophils, lymphocytes, monocytes, platelets, prothrombin time, activated partial thromboplastin time, and fibrinogen). Fifty age and sex-matched healthy controls were also evaluated. SC5b-9 and C5a plasma levels were significantly increased in the hospitalized patients (moderate and severe) in comparison with the non-hospitalized mild group. SC5b9 and C5a plasma levels were predictive of the disease severity evaluated one month later. IL-6, IL-8, TNFα, IL-10 and complement split products were higher in moderate/severe versus non-hospitalized mild COVID-19 patients and healthy controls but with a huge heterogeneity. SC5b-9 and C5a plasma levels correlated positively with CRP, ferritin values and the neutrophil/lymphocyte ratio. Complement can be activated in the very early phases of the disease, even in mild non-hospitalized patients. Complement activation can be observed even when pro-inflammatory cytokines are not increased, and predicts a negative outcome

Elevated NET, Calprotectin, and Neopterin Levels Discriminate between Disease Activity in COVID-19, as Evidenced by Need for Hospitalization among Patients in Northern Italy

Coronavirus disease 2019 (COVID-19) displays clinical heterogeneity, but little information is available for patients with mild or very early disease. We aimed to characterize biomarkers that are useful for discriminating the hospitalization risk in a COVID-19 cohort from Northern Italy during the first pandemic wave. We enrolled and followed for four weeks 76 symptomatic SARS-CoV-2 positive patients and age/sex-matched healthy controls. Patients with mild disease were discharged (n.42), and the remaining patients were hospitalized (n.34). Blood was collected before any anti-inflammatory/immunosuppressive therapy and assessed for soluble C5b-9/C5a, H3-neutrophil extracellular traps (NETs), calprotectin, and DNase plasma levels via ELISA and a panel of proinflammatory cytokines via ELLA. Calprotectin and NET levels discriminate between hospitalized and non-hospitalized patients, while DNase negatively correlates with NET levels; there are positive correlations between calprotectin and both NET and neopterin levels. Neopterin levels increase in patients at the beginning of the disease and do so more in hospitalized than non-hospitalized patients. C5a and sC5b-9, and other acute phase proteins, correlate with neopterin, calprotectin, and DNase. Both NET and neopterin levels negatively correlate with platelet count. We show that calprotectin, NETs, and neopterin are important proinflammatory parameters potentially useful for discriminating between COVID-19 patients at risk of hospitalization