Interplay between inflammation, autoimmunity and regeneration in the NOD mouse model of type 1 diabetes and Sjogren’s Syndrome.

PhDA continuous process of tissue remodelling and regeneration is a fundamental feature of the homeostatic response of the target organ of several autoimmune diseases. In type 1 diabetes (T1D) the β cell mass is in a constant process of death and renewal in order to regenerate the islets damaged by the autoimmune process. The relationship linking inflammation and regeneration during autoimmunity remains elusive. Reg genes, a multigene family discovered using cDNA libraries derived from rat regenerating islets, have been suggested to play an important role in epithelial regeneration not only in the pancreas but also in the salivary glands (SG) of Sjogren’s Syndrome (SS) during autoimmune sialoadenitis. Both in patients and animal models of T1D and SS, the chronic inflammatory/autoimmune process is heterogeneous and display high immunological variability. In particular, in a sizeable subset of cases, inflammatory lesions display ectopic lymphoid structures (ELS) characterised by T/B cell segregation, follicular dendritic cells networks and differentiation of germinal center B cells. However, there is limited evidence on the cellular and molecular mechanisms underlying ELS formation and their contribution to autoimmunity in the pancreas during autoimmune insulitis and in SG during autoimmune sialoadenitis. In this PhD project, I used the NOD mouse model of T1D and SS in order to investigate i) the cellular and molecular mechanisms regulating ELS formation, ii) the functionality of ELS in supporting in situ autoreactive B cell differentiation and iii) the relationship between formation of ELS and the expression of REG genes. In this work I showed that ELS formation was preceded by local up-regulation of lymphotoxins (LTαβ) and lymphoid chemokines CXCL13 and CCL19 and that, once formed, ELS were fully functional in promoting autoreactive B cell activation. Importantly, inhibition of the LT-β pathway prevented the formation of ELS and B cell autoimmunity. Finally, I showed that the expression pattern of Reg genes was strictly related to the development of inflammatory infiltrates in NOD 7 mice and that Reg proteins were target of the autoimmune process itself, as shown by the development of anti-Reg1 antibodies in patients with T1D. Overall, these results suggest that the processes of destruction and regeneration occurring in chronic autoimmune/inflammatory diseases are strongly interdependent whereby autoimmunity may be further enhanced by the attempt to regenerate

Lo sfondamento dell’orizzonte tradizionale: dalla prospettiva nazionale a quella globale. Stati e confessioni religiose alla prova. Religione e confessioni nell’Unione europea tra speranze ...

Il contributo, non sottoposto a valutazione, riproduce il testo integrale, corredato delle note, della relazione al Convegno Nazionale dell’ADEC sul tema “Per una disciplina che cambia. Il diritto canonico e il diritto ecclesiastico nel tempo presente” (Bologna, 7-9 novembre 2013), ed è destinato alla pubblicazione negli Atti.SOMMARIO: 1. Osservazioni introduttive - 2. Il fenomeno religioso nelle costituzioni europee del XXI secolo - 3. La bilateralità con particolare riferimento alla Germania - 4. Osservazioni conclusive. ABSTRACT This essay examines the legal condition of religious denominations in the European States’ new Constitutions. It focuses upon the legal provisions concerning the principle of cooperation and its application contained both in these Constitutions and in the German Länder’s agreements signed with religious denominations after the Reunification. The inconsistencies thereby originated are also taken into account as regards both European legal rules and the condition of religious denominations within national States

Adherence in Rheumatoid Arthritis patients assessed with a validated Italian version of the 5-item compliance questionnaire for rheumatology

OBJECTIVES: The 5-item Compliance Questionnaire for Rheumatology (CQR5) proved reliability and validity in respect of identification of patients likely to be high adherers (HAs) to anti-rheumatic treatment, or low adherers (LAs), i.e. taking<80% of their medications correctly. The objective of the study was to validate an Italian version of CQR5 (I-CQR5) in rheumatoid arthritis (RA) patients and to investigate factors associated with high adherence. METHODS: RA patients, undergoing treatment with ≥1 self-administered conventional synthetic disease-modifying anti-rheumatic drug (csDMARD) or biological DMARD (bDMARD), were enrolled. The cross-cultural adaptation and validation of I-CQR5 followed standardised guidelines. I-CQR5 was completed by patients on one occasion. Data were subjected to factor analysis and Partial Credit model Parametrisation (PCM) to assess construct validity of I-CQR5. Analysis of factors associated with high adherence included demographic, social, clinical and treatment information. Factors achieving a p<0.10 in univariate analysis were included in multivariable analysis. RESULTS: Among 604 RA patients, 274 patients were included in the validation and 328 in the analysis of factors associated with adherence. Factor analysis and PCM confirmed the construct validity and consistency of I-CQR5. HAs were found to be 109 (35.2%) of the patients. bDMARD treatment and employment were found to be independently associated with high adherence: OR 2.88 (1.36-6.1), p=0.006 and OR 2.36 (1.21-4.62), p=0.012, respectively. CONCLUSIONS: Only one-third of RA patients were HAs according to I-CQR5. bDMARDs and employment status increased by almost 3-fold the likelihood of being highly adherent to the anti-rheumatic treatment.Peer reviewe

In vitro digestibility of field pea as influenced by processing methods

Field pea meals exposed to different treatments (flaking, extrusion, expansion, dry heating at 150°C/15' or 30', dry heating at 150°C/30' after addition of 1% of xylose, 4% NaOH addition, microwave irradiation at 800 W for 6' or 9') were controlled for their 6 and 24 hours in vitro fermentability by the gas production (GP) technique. Flaking and extrusion accelerated initial fermentation but tended to reduce 24h GP, whereas dry heating and microwaves mainly improved final gas volume, but NaOH had the opposite effect. Apparent dry matter digestion at 6h was lowered by dry heating, NaOH addition and the shorter microwave irradiation. Xylose addition did not substantially change the effects of dry heating, but lowered the initial disappearance. Ammonia concentration was in general lowered by the treatments, suggesting a reduction in protein degradability but also a possible higher microbial uptake for protein synthesis. Microwave irradiation had limited effects on all the parameters. Dry heating, with or without xylose addition, seems interesting to increase rumen escaping protein fraction without accelerating starch fermentation that could expose to higher risks of rumen acidosis

CD4 T lymphocyte autophagy is upregulated in the salivary glands of primary Sjögren’s syndrome patients and correlates with focus score and disease activity

Background: Primary Sjögren’s syndrome (pSS) is a common chronic autoimmune disease characterized by lymphocytic infiltration of exocrine glands and peripheral lymphocyte perturbation. In the current study, we aimed to investigate the possible pathogenic implication of autophagy in T lymphocytes in patients with pSS. Methods: Thirty consecutive pSS patients were recruited together with 20 patients affected by sicca syndrome a nd/or chronic sialoadenitis and 30 healthy controls. Disease activity and damage were evaluated according to SS disease activity index, EULAR SS disease activity index, and SS disease damage index. T lymphocytes were analyzed for the expression of autophagy-specific markers by biochemical, molecular, and histological assays in peripheral blood and labial gland biopsies. Serum interleukin (IL)-23 and IL-21 levels were quantified by enzyme-linked immunosorbent assay. Results: Our study provides evidence for the first time that autophagy is upregulated in CD4+ T lymphocyte salivary glands from pSS patients. Furthermore, a statistically significant correlation was detected between lymphocyte autophagy levels, disease activity, and damage indexes. We also found a positive correlation between autophagy enhancement and the increased salivary gland expression of IL-21 and IL-23, providing a further link between innate and adaptive immune responses in pSS. Conclusions: These findings suggest that CD4+ T lymphocyte autophagy could play a key role in pSS pathogenesis. Additionally, our data highlight the potential exploitation of T cell autophagy as a biomarker of disease activity and provide new ground to verify the therapeutic implications of autophagy as an innovative drug target in pSS

Atrial fibrillation pattern, left atrial diameter and risk of cardiovascular events and mortality. A prospective multicenter cohort study.

BACKGROUND There are conflicting evidence on the association between atrial fibrillation (AF) pattern, such as persistent/permanent (Pers/Perm) and paroxysmal (PAF) AF and risk of ischemic events. We investigated if left atrial diameter (LAd) may affect the risk of cardiovascular outcomes according to AF pattern. METHODS Prospective multicenter observational including 1,252 non-valvular AF patients (533 PAF and 719 Pers/Perm AF). Study endpoints were cardiovascular events (CVEs), major adverse cardiac events (MACE) and CV death. LA anteroposterior diameter (LAd) was obtained by transthoracic echocardiography. RESULTS Pers/Perm AF patients had a higher proportion of LAd above median than PAF (≥44 mm, 59.5% vs 37.5% respectively, P < .001). In a mean follow-up of 42.2 ± 31.0 months (4,315 patients/year) 179 CVEs (incidence rate [IR] 4.2%/year), 133 MACE (IR 3.1%/year), and 97 CV deaths (IR 2.2%/year) occurred. Compared to patients with LAd below median, those with LAd above the median had a higher rate of CVEs (log-rank test, P < .001), MACE (log-rank test P < .001), and CV death (log-rank test P < .001). Multivariable Cox regression analysis showed that LAd above the median was associated with CVEs, (HR 1.569, 95% CI 1.129-2.180, P = .007) MACE (HR 1.858, 95% CI 1.257-2.745, P = .002) and CV death (HR 2.106, 95% CI 1.308-3.390, P = .002). The association between LAd and outcomes was evident both in PAF and Pers/Perm AF patients. No association between AF pattern and outcomes was found. CONCLUSION LAd is a simple parameter that can be obtained in virtually all AF patients and can provide prognostic information on the risk of CVEs, MACE and CV death regardless of AF pattern

Immunopathogenesis of rheumatoid arthritis

Rheumatoid arthritis (RA) is the most common inflammatory arthropathy. The majority of evidence, derived from genetics, tissue analyses, models, and clinical studies, points to an immune-mediated etiology associated with stromal tissue dysregulation that together propogate chronic inflammation and articular destruction. A pre-RA phase lasting months to years may be characterized by the presence of circulating autoantibodies, increasing concentration and range of inflammatory cytokines and chemokines, and altered metabolism. Clinical disease onset comprises synovitis and systemic comorbidities affecting the vasculature, metabolism, and bone. Targeted immune therapeutics and aggressive treatment strategies have substantially improved clinical outcomes and informed pathogenetic understanding, but no cure as yet exists. Herein we review recent data that support intriguing models of disease pathogenesis. They allude to the possibility of restoration of immunologic homeostasis and thus a state of tolerance associated with drug-free remission. This target represents a bold vision for the future of RA therapeutics

Standardisation of labial salivary gland histopathology in clinical trials in primary Sjögren's syndrome

Labial salivary gland (LSG) biopsy is used in the classification of primary Sjögren's syndrome (PSS) and in patient stratification in clinical trials. It may also function as a biomarker. The acquisition of tissue and histological interpretation is variable and needs to be standardised for use in clinical trials. A modified European League Against Rheumatism consensus guideline development strategy was used. The steering committee of the ad hoc working group identified key outstanding points of variability in LSG acquisition and analysis. A 2-day workshop was held to develop consensus where possible and identify points where further discussion/data was needed. These points were reviewed by a subgroup of experts on PSS histopathology and then circulated via an online survey to 50 stakeholder experts consisting of rheumatologists, histopathologists and oral medicine specialists, to assess level of agreement (0–10 scale) and comments. Criteria for agreement were a mean score ≥6/10 and 75% of respondents scoring ≥6/10. Thirty-nine (78%) experts responded and 16 points met criteria for agreement. These points are focused on tissue requirements, identification of the characteristic focal lymphocytic sialadenitis, calculation of the focus score, identification of germinal centres, assessment of the area of leucocyte infiltration, reporting standards and use of prestudy samples for clinical trials. We provide standardised consensus guidance for the use of labial salivary gland histopathology in the classification of PSS and in clinical trials and identify areas where further research is required to achieve evidence-based consensus

Cytokines and inflammatory mediators: 25. Certolizumab Pegol has a Different Profile from the other Anti-TNFS, Including Golimumab, in a Variety of in Vitro Assays

Background: Activities of the anti-TNFs, certolizumab pegol (CZP), etanercept (ETA), infliximab (IFX) and adalimumab (ADA), have been compared in a range of in vitro assays. CZP is the only licensed PEGylated Fab' anti-TNF; ETA is a fusion protein with an IgG1 Fc, and IFX and ADA are both antibodies with an IgG1 Fc. Golimumab (GLM) is a monoclonal IgG1 TNF inhibitor recently approved for a number of indications; it is thus of interest to assess the in vitro activity of GLM. In vitro assays previously used were neutralisation of TNF in the L929 bioassay, inhibition of LPS-driven cytokine production by monocytes, induction of apoptosis in activated lymphocytes and monocytes, and induction of neutrophil necrosis. Methods: Neutralisation of human TNF was assessed in the L929 bioassay using a range of concentrations of the anti-TNFs and a fixed concentration of TNF (100 pg/mL). Activity of the anti-TNFs at inhibiting LPS-driven IL-1β secretion by monocytes was assessed by incubating peripheral blood monocytes with various concentrations of the anti-TNF for 1 hour (hr) and then washing the cells. LPS was added for 4 hrs, the supernatants collected and the IL-1β level measured by ELISA. To assess induction of apoptosis, peripheral blood lymphocytes were activated for 2 days with 2 μg/mL CD3/CD28 and monocytes with 300 U/mL IL-4 and GMCSF for 3 days. The effect of the anti-TNFs on apoptosis was assessed by Annexin V staining using flow cytometry 24 hrs later. The effect of the anti-TNFs on neutrophil necrosis was determined by measuring myeloperoxidase release after 12 hrs. An isotype-matched control was used in all assays except the L929 bioassay. Results: IC90 neutralisation activity of the anti-TNFs in the L929 bioassay was 0.3 ng/mL for ETA, 4 ng/mL for GLM, 15 ng/mL for ADA, and 20 ng/mL for IFX, compared with 2.5 ng/mL for CZP. CZP was the most potent inhibitor of LPS-driven IL-1β secretion (IC50 ∼0.1 ng/mL), followed by GLM (20 ng/mL) and IFX (50 ng/mL). GLM, ADA, IFX and ETA induced apoptosis of monocytes and lymphocytes to a similar degree reaching a level of 23% and ∼40% at 100 μg/mL, respectively. CZP caused no increase in apoptosis above the levels seen with the isotype-matched control. In the neutrophil necrosis assay, ADA,IFX and GLM caused ∼70% necrosis at 100 μg/mL, and ETA 48%. CZP did not increase the level of necrosis above the level of the control. Conclusions: Bioactivity of the IgG1 molecules GLM, IFX and ADA in neutralising human TNF was inferior to that of CZP and ETA. CZP, the only PEGylated anti-TNF, had a different profile to the other anti-TNFs as it was the most potent at inhibiting LPS-driven IL-1β production by monocytes, did not induce apoptosis of activated monocytes and lymphocytes, and did not cause neutrophil necrosis. The clinical relevance of these in vitro effects is unknown. Nevertheless, these assays show interesting in vitro differences between the anti-TNFs. Disclosure statement: G.F. and A.N. are employees of UC