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Objectives: Pneumocystis jirovecii is an opportunistic fungus. Pneumocystis jirovecii pneumonia (PJP) is well known in the human immunodeficiency virus (HIV)-infected population, but in non-HIV-related immunosuppressed patients, risk factors are largely unknown. We studied the characteristics and outcome of patients treated for systemic autoimmune disorders infected with P. jirovecii, aiming to clarify risk stratification to guide prophylaxis. Method: Clinical charts collected between 2010 and 2016 at the University Hospital of Leuven (Belgium) were reviewed. Information on type of systemic disorder, organ involvement, immunosuppressant use, and comorbidity was collected, and laboratory results were consulted. Results: In total, 39 cases of non-HIV PJP were retrieved, 24 of whom had pre-existing pulmonary disease. All were on immunosuppressant medication at the time of infection, the majority (36/39) taking glucocorticoids, with a median dose of 16 mg methylprednisolone over the past 3 months. Of the 39 cases, 21 were admitted to the intensive care unit and mortality reached 35%. Age and pulmonary disease correlated positively and methotrexate use negatively with mortality. When applying current prophylactic strategies to our cohort, 50% of infections could theoretically have been prevented. Conclusion: PJP is a rare but relevant clinical problem when caring for immunosuppressed patients with autoimmune systemic disorders. Pulmonary disease and age are risk factors for acquiring the infection and carry a worse prognosis. More studies are needed to further define prophylactic criteria.

To study the prognostic value of myocardial native T1 and extracellular volume (ECV), measured by cardiovascular magnetic resonance (CMR), in patients with systemic sclerosis (SSc).Thirty-three SSc patients (16/33 male, 48.5%) were studied using multiparametric CMR including native T1 mapping with ECV calculation, T2 mapping, and late gadolinium enhancement (LGE). Patients were followed-up for cardiac death, haemodynamically significant arrhythmia, or heart failure. Results were compared with 33 age- and gender-matched healthy controls.When compared with controls, SSc patients had higher myocardial native T1 (1,058.9±71 versus 989.4±21.4 ms, p0.001), higher T2 (54.9±5.7 versus 50±2.5 ms, p0.001), and ECV values (27.9±5.4% versus 24.8±2%, p0.004). LGE was present in eight patients (24%), two subendocardial, five midwall, and four subepicardial. LGE, native T1, and ECV were significantly associated with adverse events during follow-up in multivariate Cox regression analysis. Kaplan-Meier analysis demonstrated significant divergence of the survival curves based on the presence of elevated native T1 (≥1,069 ms) or ECV (≥31.4%) values.Cardiac involvement is frequent in SSc. Both native T1 mapping and ECV represent novel non-invasive markers of myocardial fibrosis and could be used in the risk stratification of patients with SSc. CMR mapping may provide a novel biomarker for disease monitoring and study of therapies aiming to reduce myocardial fibrosis in SSc.

Background:RISE-SSc (NCT02283762) was a multicenter, double-blind, Phase IIb study of riociguat in early dcSSc. Primary endpoint was change in mRSS from baseline to Wk 52.Objectives:Exploratory, descriptive analyses of riociguat target engagement and effects on disease biomarkers in RISE-SSc and their relationship with effects on the primary endpoint. All biomarker p-values are for information only.Methods:Pts with dcSSc (duration ≤18 mo; modified Rodnan skin score [mRSS] 10–22 units) were randomized to riociguat 0.5−2.5 mg tid (n=60) or placebo (n=61). Biomarkers of target engagement (cGMP), inflammation and/or vascular/endothelial function (e.g. high-sensitivity C-reactive protein [hsCRP], soluble platelet endothelial cell adhesion molecule 1 [sPECAM-1], soluble E-selectin, chemokine ligand 4 [CXCL-4]), and fibrosis (e.g. alpha-smooth muscle cell actin [alphaSMA], pro-collagen mRNA expression) were measured in plasma, serum, and skin biopsies at baseline and Wk 14.Results:Mean±SD change from baseline in mRSS was –2.09±5.66 (n=57) with riociguat and –0.77±8.24 (n=52) with placebo (p=0.08). From baseline to Wk 14, plasma cGMP rose by mean (SD) 94% (78%) (n=52) with riociguat and 10% (39%) (n=52) with placebo (nominal pConclusion:Primary study endpoint (change in mRSS) was not met. Plasma cGMP rose with riociguat, confirming engagement with the NO-sGC-cGMP pathway. Serum sPECAM-1 (marker of endothelial activation) and CXCL-4 (marker of progressive SSc) fell with riociguat; hsCRP and E-selectin did not. Some serum and skin biomarkers of higher disease activity at baseline were associated with a greater effect of riociguat on skin fibrosis.Acknowledgments:RISE-SSc was jointly funded by Bayer AG and Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc., Kenilworth, NJ, USA.Disclosure of Interests:Oliver Distler Grant/research support from: Grants/Research support from Actelion, Bayer, Boehringer Ingelheim, Competitive Drug Development International Ltd. and Mitsubishi Tanabe; he also holds the issued Patent on mir-29 for the treatment of systemic sclerosis (US8247389, EP2331143)., Consultant of: Consultancy fees from Actelion, Acceleron Pharma, AnaMar, Bayer, Baecon Discovery, Blade Therapeutics, Boehringer, CSL Behring, Catenion, ChemomAb, Curzion Pharmaceuticals, Ergonex, Galapagos NV, GSK, Glenmark Pharmaceuticals, Inventiva, Italfarmaco, iQvia, medac, Medscape, Mitsubishi Tanabe Pharma, MSD, Roche, Sanofi and UCB, Speakers bureau: Speaker fees from Actelion, Bayer, Boehringer Ingelheim, Medscape, Pfizer and Roche, Frank Kramer Employee of: Bayer AG, Josef Höfler Employee of: Josef Höfler is an employee of Staburo GmbH, Munich, Germany, contracted by Bayer AG to perform the biomarker analyses, Mercedeh Ghadessi Employee of: Bayer AG, Peter Sandner Employee of: Bayer AG, Yannick Allanore Grant/research support from: BMS, Inventiva, Roche, Sanofi, Consultant of: Actelion, Bayer AG, BMS, BI, Christopher Denton Grant/research support from: GlaxoSmithKline, CSL Behring, and Inventiva, Consultant of: Medscape, Roche-Genentech, Actelion, GlaxoSmithKline, Sanofi Aventis, Inventiva, CSL Behring, Boehringer Ingelheim, Corbus Pharmaceuticals, Acceleron, Curzion and Bayer, Masataka Kuwana Grant/research support from: Acetelion, Consultant of: Acetelion, Bayer, Chugai, Corbus Pharmaceuticals, CSL Behring and Reata Pharmaceuticals. He was a member of the SENSCIS trial Steering Committee (Boehringer Ingelheim), Marco Matucci-Cerinic Grant/research support from: Actelion, MSD, Bristol-Myers Squibb, Speakers bureau: Acetelion, Lilly, Boehringer Ingelheim, Janet Pope Grant/research support from: AbbVie, Bristol-Myers Squibb, Eli Lilly & Company, Merck, Roche, Seattle Genetics, UCB, Consultant of: AbbVie, Actelion, Amgen, Bayer, Boehringer Ingelheim, Bristol-Myers Squibb, Eicos Sciences, Eli Lilly & Company, Emerald, Gilead Sciences, Inc., Janssen, Merck, Novartis, Pfizer, Roche, Sandoz, Sanofi, UCB, Speakers bureau: UCB, Tatsuya Atsumi Grant/research support from: Eli Lily Japan K.K., Alexion Pharmaceuticals, Inc., Bristol-Myers Squibb Co., AbbVie Inc., Daiichi Sankyo Co., Ltd., Pfizer Inc., Chugai Pharmaceutical Co., Ltd., Mitsubishi Tanabe Pharma Co., Astellas Pharma Inc., Consultant of: Gilead Sciences, Inc., Eli Lilly Japan K.K., UCB Japan Co. Ltd., AbbVie Inc., Daiichi Sankyo Co., Ltd., Pfizer Inc., Chugai Pharmaceutical Co., Ltd., Speakers bureau: Eli Lilly Japan K.K., UCB Japan Co. Ltd., Bristol-Myers Squibb Co., AbbVie Inc., Eisai Co. Ltd., Otsuka Pharmaceutical Co., Ltd., Daiichi Sankyo Co., Ltd., Pfizer Inc., Chugai Pharmaceutical Co., Ltd., Mitsubishi Tanabe Pharma Co., Takeda Pharmaceutical Co., Ltd., Astellas Pharma Inc., Radim Bečvář Consultant of: Actelion, Roche, László Czirják Consultant of: Actelion, BI, Roche-Genentech, Lilly, Medac, Novartis, Pfizer, Bayer AG, Ellen De Langhe Consultant of: member of advisory board for Boehringer, Eric Hachlla: None declared, Tomonori Ishii: None declared, Osamu Ishikawa: None declared, Sindhu Johnson Grant/research support from: Boehringer Ingelheim, Corbus Pharmaceuticals, GlaxoSmithKline, Roche, Merck, Bayer, Consultant of: Boehringer Ingelheim, Ikaria, Kaisa Laapas Employee of: Partly in-sourced to Bayer, Valeria Riccieri: None declared, Elena Schiopu: None declared, Richard Silver: None declared, Vanessa Smith Grant/research support from: The affiliated company received grants from Research Foundation - Flanders (FWO), Belgian Fund for Scientific Research in Rheumatic diseases (FWRO), Boehringer Ingelheim Pharma GmbH & Co and Janssen-Cilag NV, Consultant of: Boehringer-Ingelheim Pharma GmbH & Co, Speakers bureau: Actelion Pharmaceuticals Ltd, Boehringer-Ingelheim Pharma GmbH & Co and UCB Biopharma Sprl, Chiara Stagnaro: None declared, Viginia Steen Grant/research support from: The associated affiliation has received grants/research from Boehringer Ingelheim, Corbus Pharmaceuticals, CSL Behring, Eicos, Galapagos, Immune Tolerance Network, Reata, Consultant of: Virginia Steen has acted as a consultant for Boehringer Ingelheim, Corbus, CSL Behring, Eicos, Forbius, Wendy Stevens: None declared, Gabriella Szücs: None declared, Marie-Elise Truchetet: None declared, Melanie Wosnitza Employee of: Bayer AG, Dinesh Khanna Shareholder of: Eicos Sciences, Inc./Civi Biopharma, Inc., Grant/research support from: Dr Khanna was supported by NIH/NIAMS K24AR063120, Consultant of: Acceleron, Actelion, Bayer, Boehringer Ingelheim, Bristol-Myers Squibb, Corbus Pharmaceuticals, Horizon Therapeutic, Galapagos, Roche/Genentech, GlaxoSmithKline, Mitsubishi Tanabe, Sanofi-Aventis/Genzyme, UCB

Objectives: This study, developed within the Innovative Medicines Initiative Joint Undertaking project PRECISESADS framework, aimed at functionally characterize the monocyte subsets in RA patients, and analyze their involvement in the increased CV risk associated with RA. Methods: The frequencies of monocyte subpopulations in the peripheral blood of 140 RA patients and 145 healthy donors (HDs) included in the PRECISESADS study were determined by flow cytometry. A second cohort of 50 RA patients and 30 HDs was included, of which CD14+ and CD16+ monocyte subpopulations were isolated using immuno-magnetic selection. Their transcriptomic profiles (mRNA and microRNA), proinflammatory patterns and activated pathways were evaluated and related to clinical features and CV risk. Mechanistic in vitro analyses were further performed. Results: CD14++CD16+ intermediate monocytes were extended in both cohorts of RA patients. Their increased frequency was associated with the positivity for autoantibodies, disease duration, inflammation, endothelial dysfunction and the presence of atheroma plaques, as well as with the CV risk score. CD14+ and CD16+ monocyte subsets showed distinctive and specific mRNA and microRNA profiles, along with specific intracellular signaling activation, indicating different functionalities. Moreover, that specific molecular profiles were interrelated and associated to atherosclerosis development and increased CV risk in RA patients. In vitro, RA serum promoted differentiation of CD14+CD16- to CD14++CD16+ monocytes. Co-culture with RA-isolated monocyte subsets induced differential activation of endothelial cells. Conclusions: Our overall data suggest that the generation of inflammatory monocytes is associated to the autoimmune/inflammatory response that mediates RA. These monocyte subsets, -which display specific and distinctive molecular signatures- might promote endothelial dysfunction and in turn, the progression of atherosclerosis through a finely regulated process driving CVD development in RA. ispartof: FRONTIERS IN IMMUNOLOGY vol:10 ispartof: location:Switzerland status: published

Background:The presence of myositis specific anti-melanoma differentiation associated protein 5 (MDA5) autoantibodies is associated with mucocutaneous ulcerations, rapidly progressing interstitial lung disease (RPILD), arthritis and mild muscle involvement in patients. RPILD is the major cause of mortality. At present it is unknown which domain of the MDA5 protein is the main elicitor of an immunogenic response.Objectives:The aim of this study is to delineate the domains in the MDA5 protein that are the target of autoantibodies.Methods:Anti-MDA5 IgG were isolated from MDA5(+) patient plasma (7 UPMC, 1 KI and 1 KULeuven) by affinity chromatography using an in-house affinity column as described earlier in Ossipova et al, 2014(1). 8 constructs covering different regions of the MDA5 protein were recombinantly produced in E.coli (Uniprot ID Q9BYX4, Figure 1). An in-house ELISA was developed to identify the domains with the main epitope(s) by measuring the reactivity of the plasma samples and purified autoantibodies against these MDA5 protein constructs, similar to what was reported by Fernandes-Cerqueira et al, 2018(2). The biotinylated MDA5 proteins were immobilized on streptavidin coated plates and subsequently incubated with primary antibodies (purified autoantibodies(2) or original plasma) and a HRP-conjugated secondary antibody. The ELISA was developed by the addition of TMB substrate and the optical density (OD) was measured at 450 nm.Figure 1.Graphical presentation of the constructs representing different (combinations of) domains of the MDA5 protein.Results:The preliminary data suggest the main reactivity of the plasma samples and the corresponding purified autoantibodies is directed towards the helicase domains and that there is variability between the patients in the reactivity towards domains located at the end of the protein.Conclusion:The study aims to resolve the main immunogenic domain of the MDA5 protein, which will lead to more insight in the disease mechanisms. The preliminary results suggest this domain is in the center of the MDA5 protein, but further experiments are necessary. We will use this set up to study differences in reactivity between patients (from different cohorts) and assess if differences in antibody reactivity could be linked to clinical features such as RPILD. Such correlations might be beneficial to predict the disease progression and to apply personal treatment approaches.References:[1]Ossipova E, Cerqueira CF, Reed E, Kharlamova N, Israelsson L, et al. Affinity purified anti-citrullinated protein/peptide antibodies target antigens expressed in the rheumatoid joint. Arthritis Res Ther. 2014;16(4):R167.[2]Fernandes-Cerqueira C, Renard N, Notarnicola A, Wigren E, Gräslund S, et al. Patients with anti-Jo1 antibodies display a characteristic IgG Fc-glycan profile which is further enhanced in anti-Jo1 autoantibodies. Scientific reports. 2018;8(1):17958.Disclosure of Interests:Eveline Van Gompel: None declared, Catia Cerqueira: None declared, Edvard Wigren: None declared, Susanne Gräslund: None declared, Karine Chemin: None declared, Begum Horuluoglu: None declared, Ellen De Langhe: None declared, Olivier Benveniste: None declared, Ingrid E. Lundberg Consultant of: Consulting fees from Corbus Pharmaceuticals, Inc, Grant/research support from: Research grants from Bristol Myers Squibb and AstraZeneca.

Career situation of first and presenting author Student for a master or a PhD. Introduction The role of epigenetic factors in the pathophysiology of fibrosis, a hallmark of Systemic Sclerosis, is increasingly explored. DOT1L, the unique H3K79-methyltransferase, methylates histone 3 at the Lysine residue at position 79, thereby regulating gene expression programs. In cartilage and bone, DOT1L has cell-type specific effects on Wnt signaling, a pathway suggested to play an important role in fibrosis. Objectives To study the role of DOT1L in fibrosis. Methods Primary human dermal fibroblasts were treated with DOT1L-inhibitor EPZ-5676 or vehicle and stimulated with TGF-β. Expression of smooth muscle alpha 2 actin (ACTA2) and Wnt target genes was measured by RT-qPCR. Western Blot was done for dimethylated H3K79 and β-catenin. Picrosirius Red staining measured collagen deposition. 5-Bromo-2’-deoxy-uridine (BrdU) labeling for proliferation and flow cytometry with Propidium Iodide for cell cycle analysis was done. Col1a2;Cre-ERT2;DOT1lfl/flmice, injected with tamoxifen to induce a fibroblast-specific DOT1L knockout, were injected subcutaneously with bleomycin or vehicle. Injected skin was analyzed by OH-prolin assay and by histology. Results The DOT1L-inhibitor EPZ-5676 reduced H3K79 dimethylation in all samples. In breast dermal fibroblasts, the induction of ACTA2 with TGF-β was reduced with DOT1L inhibition, while in abdominal dermal fibroblasts this induction was more pronounced. After 48 hours of TGF-β, collagen deposition was higher in DOT1L-inhibited fibroblasts. After 72 hours of TGF-β however, this deposition was comparable with controls. DOT1L inhibition induced canonical Wnt signaling in fibroblasts, with a small increase in active β-catenin and expression of Lymphoid enhancer-binding factor 1 (LEF1). With DOT1L inhibition, more proliferation of fibroblasts, but also proportionally more cells in the G1/G0 phase and less cells in S and M/G2 phase were seen. In vivo, subcutaneous bleomycin increased murine dermal thickness and skin collagen content. No difference was observed between wild type and mice with a fibroblast-specific deletion of DOT1L. Conclusions In an in vitro model of fibrosis, ACTA2 induction in DOT1L-inhibited human dermal fibroblasts was dependent on the fibroblast origin. DOT1L inhibition resulted in an earlier deposition of collagen, without differences in deposition at the end points. Inhibition of DOT1L induced canonical Wnt signaling and proliferation but also led to a higher proportion of cells in the G0/G1 phase. In an in vivo murine model of skin fibrosis, no difference in bleomycin-induced skin thickness and collagen content was found when the DOT1L gene was deleted in fibroblasts. Disclosure of Interest None declared.

Background:Systemic sclerosis-associated interstitial lung disease (SSc-ILD) is the main cause of death in SSc and accounts for up to 30-35% of SSc-mortality (1-2). All SSc cases, irrespective of the extent of the skin disease, should be evaluated for ILD (3). The epidemiology of SSc-ILD in Belgium is unknown. In literature, the prevalence of ILD in SSc varies between 19% and 52%. However, different criteria were used to diagnose ILD (4). In 2008, Goh et al. proposed a flow diagram to diagnose SSc-ILD based on chest high-resolution CT-scan (HRCT) and pulmonary function tests (PFTs). Their categorization into limited or extensive ILD has prognostic value (5).Objectives:To determine the prevalence and incidence rate of SSc-ILD in Flanders.Methods:Up to 12-year follow-up data of consecutive SSc patients were obtained by 2 Flemish expert centres (University Hospitals Ghent and Leuven). Patients fulfilling the LeRoy and/or ACR-EULAR classification criteria were included consecutively in the prospective cohort (6). Patients received HRCT at baseline and on indication thereafter, as well as yearly PFT. All HRCTs were centrally analyzed (Ghent) and patients were categorized according to the Goh criteria as without ILD, with limited ILD (limILD) or with extensive ILD (extILD) (5).Results:Between 2006 and 2018, 797 SSc patients (557 Ghent/240 Leuven; 22% limited SSc (LSSc)/59% limited cutaneous SSc (LcSSc)/19% diffuse cutaneous SSc (DcSSc)) had baseline HRCT and PFT. The baseline characteristics are depicted in the table. The mean age (SD) was 53 +/-15 years and the majority of patients was female (76%).272 SSc patients had ILD at baseline, implicating a baseline prevalence of 34% (272/797). The baseline prevalences were 35% and 55% for the LcSSc and DcSSc subgroups respectively. During a median follow-up of 39 months (IQR: 11-79 months), 44 patients were diagnosed with incidental SSc-ILD, resulting in an incidence rate of 21,0/1000 person-years (PY), 95% CI:15,2-28,1. The incidence rates were 21,7/1000 PY, 95%CI: 14,3-31,6 and 43.9/1000PY, 95%CI: 22.7-76.8 for the LcSSc and DcSSc subgroups respectively.Table.Baseline characteristicsSSc (n=797)LcSSc (n=470)DcSSc (n=149)age (years) °53+/-1554+/-1554+/-14♂/♀ *193(24%)/604(76%)109(23%)/361(77%)58(39%)/91(61%)Disease Duration (months) #for 718: 22 (5-72)for 443: 25 (5-80)for 145: 16 (7-52)LSSc/LcSSc/DcSSc *178(22%)/470(59%)/149(19%)follow-up (months) #39 (11-79)38.5 (9.75-81)44 (17.5-78)Anti-centromere antibodies§252/538 (47%)163/317 (51%)19/108 (18%)Anti-topoisomeraseI antibodies§119/519 (23%)66/297 (22%)45/112 (40%)ILD at baseline, *272 (34%)163 (35%)82 (55%)LimILD, *230 (29%)139 (30%)67 (45%)ExtILD, *42 (5%)24 (5%)15 (10%)New ILD during follow-up, §44/52527/30712/67°: mean +/- standard deviation, *: number of patients (percent), #: median (interquartile range), §= number of patients/total number of patients with available data (%)Conclusion:In an unselected cohort of SSc patients, a third of the patients has ILD at baseline which is in line with previous prevalence reports. Importantly, this is the first study reporting incidence rates of SSc-ILD.References:[1]Steen VD and Medsger TA, Ann Rheum Dis 2007;66:940-4[2]Elhai M et al. Ann Rheum Dis 2017;76:1897-1905[3]Smith V et al. RMD Open 2019;4:e000782. doi:10.1136/rmdopen-2018-000782[4]Bergamasco A et al. Clinical Epidemiology 2019;11:257-73[5]Goh N et al. Am J Respir Crit Care Med 2008;177:1248-54[6]van den Hoogen et al. Arthritis Rheumatol 2013;65:2737-47Disclosure of Interests:Els Vandecasteele Grant/research support from: my institution has received a research grant from the Research Foundation Flanders FWO), Speakers bureau: my institution has received speaker fees from Actelion, Karin Melsens: None declared, Daniel Blockmans Consultant of: yes, Speakers bureau: yes, Charlotte Carton: None declared, Filip De Keyser: None declared, Ellen De Langhe Consultant of: member of advisory board for Boehringer, Bernard Lauwerys: None declared, Yves Piette: None declared, Amber Vanhaecke: None declared, Koen Verbeke: None declared, Wim Wuyts Grant/research support from: my institution has received a grant from Boehringer Ingelheim and Roche, Consultant of: my institution has received payments for consultancy from Boehringer Ingelheim and Roche, Speakers bureau: my institution has received speaker fees from Boehringer Ingelheim and Roche, Guy Brusselle: None declared, Vanessa Smith Grant/research support from: The affiliated company received grants from Research Foundation - Flanders (FWO), Belgian Fund for Scientific Research in Rheumatic diseases (FWRO), Boehringer Ingelheim Pharma GmbH & Co and Janssen-Cilag NV, Consultant of: Boehringer-Ingelheim Pharma GmbH & Co, Speakers bureau: Actelion Pharmaceuticals Ltd, Boehringer-Ingelheim Pharma GmbH & Co and UCB Biopharma Sprl

Background:RISE-SSc (NCT02283762) was a multicenter Phase IIb trial of riociguat in pts with early (duration ≤18 months) dcSSc and modified Rodnan skin score (mRSS) 10−22 units. Pts were randomized double-blind to placebo or riociguat 0.5–2.5 mg t.i.d. for 52 weeks. The primary endpoint, mRSS change from baseline to Week (Wk) 52, did not reach statistical significance (p=0.08, riociguat vs placebo), but there were favorable trends in some other outcomes.Objectives:To present open-label long-term extension (LTE) results of RISE-SSc.Methods:Pts who completed Wk 52 of double-blind therapy could enter LTE on riociguat. Endpoints included mRSS, adverse events (AEs), and serious AEs (SAEs).Results:Of 60 pts randomized to riociguat and 61 to placebo, 42 (riociguat−riociguat group) and 45 (former placebo group), respectively, entered LTE. At LTE start, mean±SD mRSS was 16.4±3.2 and 16.3±4.2 units, and mean disease duration was 8.9±7.8 and 8.9±5.8 months, in the riociguat−riociguat and former placebo groups, respectively. Other demographics/disease characteristics were also comparable. Median duration of riociguat treatment was 1092 d in riociguat−riociguat pts and 649 d in former placebo pts. Throughout the study, mRSS decreased in both groups (Figure 1). From Wk 52 to last visit, mRSS fell by −3.02±5.51 in riociguat−riociguat patients and −3.96±5.43 in former placebo pts. Rates of mRSS regression (decrease by >5 units and ≥25% from Wk 52 to last visit) and of % declines in mRSS were similar in the two groups (Figure 2). mRSS progression (increase by >5 units and ≥25% from Wk 52 to last visit) occurred in 1 pt (2%) in each group. During the entire study, rescue therapy agents were used in 15 (36%) riociguat−riociguat pts and 17 (38%) former placebo pts. AEs were reported from Wk 52 to last visit in 82 pts (94%): 40 (95%) riociguat−riociguat and 42 (93%) former placebo. Most common AEs overall: nasopharyngitis (24%), gastroesophageal reflux disease (17%), diarrhea (15%), and hypotension (14%). AEs of special interest (dizziness, postural dizziness, or hypotension) occurred in 5 riociguat−riociguat pts (12%) and 4 former placebo pts (9%). SAEs were reported in 21 (24%) pts: 10 (24%) riociguat−riociguat pts and 11 (24%) former placebo pts, with no SAE reported in >1 patient, no SAEs of special interest, and no deaths.Conclusion:During LTE riociguat treatment, mRSS decreased in both groups from Wk 52 onwards and mRSS progression was uncommon. Riociguat had acceptable safety, similar to the main study, with no new safety signal.Acknowledgments:RISE-SSc was jointly funded by Bayer AG and Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc., Kenilworth, NJ, USA.Disclosure of Interests:Dinesh Khanna Shareholder of: Eicos, Grant/research support from: NIH NIAID, NIH NIAMS, Consultant of: Acceleron, Actelion, Bayer, BMS, Boehringer-Ingelheim, Corbus, Galapagos, Genentech/Roche, GSK, Mitsubishi Tanabi, Sanofi-Aventis/Genzyme, UCB Pharma, Janet Pope Grant/research support from: AbbVie, Bristol-Myers Squibb, Eli Lilly & Company, Merck, Roche, Seattle Genetics, UCB, Consultant of: AbbVie, Actelion, Amgen, Bayer, Boehringer Ingelheim, Bristol-Myers Squibb, Eicos Sciences, Eli Lilly & Company, Emerald, Gilead Sciences, Inc., Janssen, Merck, Novartis, Pfizer, Roche, Sandoz, Sanofi, UCB, Speakers bureau: UCB, Marco Matucci-Cerinic Grant/research support from: Actelion, MSD, Bristol-Myers Squibb, Speakers bureau: Acetelion, Lilly, Boehringer Ingelheim, Masataka Kuwana Grant/research support from: Acetelion, Consultant of: Acetelion, Bayer, Chugai, Corbus Pharmaceuticals, CSL Behring and Reata Pharmaceuticals. He was a member of the SENSCIS trial Steering Committee (Boehringer Ingelheim), Christopher Denton Grant/research support from: GlaxoSmithKline, CSL Behring, and Inventiva, Consultant of: Medscape, Roche-Genentech, Actelion, GlaxoSmithKline, Sanofi Aventis, Inventiva, CSL Behring, Boehringer Ingelheim, Corbus Pharmaceuticals, Acceleron, Curzion and Bayer, Yannick Allanore Grant/research support from: BMS, Inventiva, Roche, Sanofi, Consultant of: Actelion, Bayer AG, BMS, BI, Melanie Wosnitza Employee of: Bayer AG, Marie-Elise Truchetet: None declared, Gabriella Szücs: None declared, Wendy Stevens: None declared, Viginia Steen Grant/research support from: The associated affiliation has received grants/research from Boehringer Ingelheim, Corbus Pharmaceuticals, CSL Behring, Eicos, Galapagos, Immune Tolerance Network, Reata, Consultant of: Virginia Steen has acted as a consultant for Boehringer Ingelheim, Corbus, CSL Behring, Eicos, Forbius, Chiara Stagnaro: None declared, Vanessa Smith Grant/research support from: The affiliated company received grants from Research Foundation - Flanders (FWO), Belgian Fund for Scientific Research in Rheumatic diseases (FWRO), Boehringer Ingelheim Pharma GmbH & Co and Janssen-Cilag NV, Consultant of: Boehringer-Ingelheim Pharma GmbH & Co, Speakers bureau: Actelion Pharmaceuticals Ltd, Boehringer-Ingelheim Pharma GmbH & Co and UCB Biopharma Sprl, Richard Silver: None declared, Elena Schiopu: None declared, Valeria Riccieri: None declared, Frank Kramer Employee of: Bayer AG, Sindhu Johnson Grant/research support from: Boehringer Ingelheim, Corbus Pharmaceuticals, GlaxoSmithKline, Roche, Merck, Bayer, Consultant of: Boehringer Ingelheim, Ikaria, Osamu Ishikawa: None declared, Tomonori Ishii: None declared, Eric Hachlla: None declared, Ellen De Langhe Consultant of: member of advisory board for Boehringer, László Czirják Consultant of: Actelion, BI, Roche-Genentech, Lilly, Medac, Novartis, Pfizer, Bayer AG, Radim Bečvář Consultant of: Actelion, Roche, Tatsuya Atsumi Grant/research support from: Eli Lily Japan K.K., Alexion Pharmaceuticals, Inc., Bristol-Myers Squibb Co., AbbVie Inc., Daiichi Sankyo Co., Ltd., Pfizer Inc., Chugai Pharmaceutical Co., Ltd., Mitsubishi Tanabe Pharma Co., Astellas Pharma Inc., Consultant of: Gilead Sciences, Inc., Eli Lilly Japan K.K., UCB Japan Co. Ltd., AbbVie Inc., Daiichi Sankyo Co., Ltd., Pfizer Inc., Chugai Pharmaceutical Co., Ltd., Speakers bureau: Eli Lilly Japan K.K., UCB Japan Co. Ltd., Bristol-Myers Squibb Co., AbbVie Inc., Eisai Co. Ltd., Otsuka Pharmaceutical Co., Ltd., Daiichi Sankyo Co., Ltd., Pfizer Inc., Chugai Pharmaceutical Co., Ltd., Mitsubishi Tanabe Pharma Co., Takeda Pharmaceutical Co., Ltd., Astellas Pharma Inc., Oliver Distler Grant/research support from: Grants/Research support from Actelion, Bayer, Boehringer Ingelheim, Competitive Drug Development International Ltd. and Mitsubishi Tanabe; he also holds the issued Patent on mir-29 for the treatment of systemic sclerosis (US8247389, EP2331143)., Consultant of: Consultancy fees from Actelion, Acceleron Pharma, AnaMar, Bayer, Baecon Discovery, Blade Therapeutics, Boehringer, CSL Behring, Catenion, ChemomAb, Curzion Pharmaceuticals, Ergonex, Galapagos NV, GSK, Glenmark Pharmaceuticals, Inventiva, Italfarmaco, iQvia, medac, Medscape, Mitsubishi Tanabe Pharma, MSD, Roche, Sanofi and UCB, Speakers bureau: Speaker fees from Actelion, Bayer, Boehringer Ingelheim, Medscape, Pfizer and Roche

Background Systemic sclerosis (SSc) is a severe chronic connective tissue disease with a high disease burden. Oral involvement with impaired oral aperture (microstomia) is frequent and associated with impaired food intake, oral hygiene and secondary dental problems. Preventive measures through mouth-stretching and oral augmentation exercises have been shown to reverse the progression of microstomia. Objectives This exploratory study assesses the effectiveness and feasibility of two different exercise approaches designed to increase oral aperture. Methods Two groups had to exercise for 10 min, 3 times/day for 3 months. Group A exercised with a passive jaw motion device (Therabite), and Group B did mouth-stretching exercises. Patients were contacted 4 times by telephone to address encountered problems. The subjects used an exercise diary to document compliance. Patients were evaluated at baseline, 3 months (period without intervention), 6 months (at the end of the treatment after 3 months of intervention) and 9 months (follow-up). Results At present, 9 patients (Therabite n=4, mouth-stretching exercises n=5) were included and recruitment is ongoing. Seven patients completed the study and increase of oral aperture was observed in all patients in both groups. In the Therabite group, after 3 months of exercise, increase of oral aperture was 9, 2, 9 and 10 mm. In the mouth-stretching exercise group the increase of oral aperture was 11, 10 and 4 mm after 3 months. The compliance, measured as the ratio of executed exercises relative to the planned number of exercises was 95,2%, 85,7%, 98,9% and 63,7% in the Therabite group and 97,4%, 48,6% and 68,3% in the mouth-stretching exercise group. Conclusions An increase of oral aperture is observed in all patients after 3 months of exercising with the Therabite device as well as after mouth-stretching exercises. No clear differences are observed between both groups, but the study was not designed nor powered for this. Remarkably, a high compliance for the treatment regime was observed in most patients. References [1] Agarwal SK. The genetics of systemic sclerosis. Discov Med. 2010Aug;10(51):134–43. [2] Alantar A, Cabane J, Hachulla E, Princ G, Glinist D, Hassin M, Sorel M, Maman L, Pilat A, Mouthon L. Recommendations for the Care of Oral Involvement in Patients With Systemic Sclerosis. Arthritis Care and Research. VOL. 63. No8, august 2011, pp 1126–1133. [3] Albilia JB, Lam DK, Blanas N, Clokie CML, Sandor GKB. Small Mouths…Big Problems? A review of Scleroderma and its Oral Health Implications. www.cda-adc.ca/jcda. November 2007, vol 73,no 9. [4] Maddali-Bongi S, Landi G, Galluccio F, Del Rosso A, Miniati I, Conforti M.L, Casale R, Matucci-Cerinic M. The rehabilitation of facial involvement in systemic sclerosis: efficacy of the combination of connective tissue massage, Kabat’s technique and kinesitherapy: a randomized controlled trial. Rheumatol Int (201131:895–901. [5] Pizzo G, Scardina GA, Mesina P. Effects of nonsurgical exercise program on the decreased mouth opening in patient with systemic sclerosis. Clin Oral Invest2003; 7:175–178. Disclosure of Interest None declared

Background The association between systemic sclerosis (SSc) and primary biliary cholangitis (PBC) is well known. However, classifications criteria of the 2 diseases have been recently revised and may change the relationships. Furthermore, the specific outcomes with regards to organ involvement and also of liver aspects have been only scarcely investigated. Objectives To describe clinical characteristics of SSc-PBC patients compared to SSc using a large series Methods A multicentre EUSTAR study collection data of SSc patients with known PBC or with PBC-specific antibodies (abs) and of SSc controls matched for disease duration and cutaneous subset. Data were recorded at baseline and at the last available visit. Results 229 patients were enrolled (85 SSc-PBC and 144 SSc). The mean age of the population at the SSc diagnosis was of 53±12.8. Baseline characteristics. The limited cutaneous subset was the most common. Anticentromere abs (ACA) were present in 82.9% of SSc-PBC patients (vs 68.5% in SSc, p-value=0.01) while anti-topoisomerase I abs were less frequently (1.2% vs 9.7%), p-value=0.02. Out of 85 SSc-PBC patients, antimitochondrial abs (AMA) were present in 80%, only 9.4% presented anti-gp210 abs and 7.1% anti-sp100 abs. The two populations did not differ for fibrosis at HRCT, lung function tests, the value of creatinine and for pulmonary arterial hypertension. A trend towards statistical significance was found in the prevalence of digital ulcers (DUs) as patients that have never suffered from past or current DUs were greater in the SSc-PBC group (78.6% vs 66%, p-value=0.051). Regarding other autoimmune associated diseases, a greater prevalence of Hashimoto thyroiditis was found in SSc-PBC (p-value=0.03). At baseline, transaminase, alkaline phosphatase and γGT levels were all higher in PBC-SSc (p-value Conclusions PBC is more present in ACA positive lcSSc; SSc-PBC patients have a higher risk of polyautoimmunity. Of the most interest, SSc-PBC patients seem to have a milder SSc phenotype with less severe organ involvement and progression. Regarding PBC phenotype, only 4 patients presented portal hypertension and nobody was subjected to liver transplantation. At baseline, the SSc-PBC group had higher cholestatic liver enzymes and more than 60% were treated with deoxycholic acid. In the future, it will be useful to evaluate the PBC phenotype during the follow-up enrolling a greater number of patients. Disclosure of Interest None declared

Introduction Fibrosis is a key process in Systemic Sclerosis, a devastating disease that is not well understood. The role of epigenetic disease mechanisms is increasingly explored. DOT1L, the unique H3K79-methyltransferase, methylates histone 3 at the Lysine residue at position 79, and regulates gene expression. Inhibition of DOT1L has cell-type specific effects on Wnt-signalling, a pathway suggested to play an important role in fibrosis. Objectives To study the role of DOT1L in fibrosis. Methods Primary cell cultures of fibroblasts, isolated from healthy human skin and treated with DOT1L-inhibitor EPZ-5676 or vehicle for 14 days, were stimulated with TGFβ 5 ng/ml after starvation. Smooth muscle alpha 2 actin (ACTA2) gene expression was measured by RT-qPCR. Efficiency of DOT1L-inhibition was analysed with Western Blot for dimethylated H3K79. Picrosirius Red staining measured collagen I and III deposition. Proliferation was analysed with 5-Bromo-2’-deoxy-uridine (BrdU) labelling. 7–9 week old Col1a2; Cre-ERT2; DOT1lfl/fl mice were injected intraperitoneally with tamoxifen (1 mg/day,5 days) to induce a fibroblast-specific DOT1L knockout. Bleomycin (0.1 mg) or vehicle was injected subcutaneously (5 days/week, 4 weeks). Injected skin was analysed by OH-prolin assay for collagen content, and by histology for dermal thickness measurement. Results The DOT1L-inhibitor EPZ-5676 reduced H3K79 dimethylation in all samples. TGFβ increased expression of ACTA2 but 3 out of 4 DOT1L inhibited samples showed a significantly reduced effect of TGFβ on ACTA2 expression. The amount of collagen I and III in the extracellular matrix after 72 hours of TGFβ, was comparable between control and EPZ treated fibroblasts. BrdU labelling assay showed increased proliferation with DOT1L inhibition. In vivo, subcutaneous bleomycin induced an increased dermal thickness and skin collagen content in mice. No difference in the effect of bleomycin was found between mice with a conditional fibroblast-specific DOT1L knockout or wild type mice. Conclusions In an in vitro model of fibrosis, primary human dermal fibroblasts treated with a DOT1L-inhibitor showed increased proliferation and reduced upregulation of ACTA2 but did not result in detectable differences in collagen deposition. In an in vivo murine model of skin fibrosis, no difference in bleomycin-induced skin thickness and collagen content was found when DOT1L was knocked out in fibroblasts. Disclosure of interest None declared

The Global Strategy for the Conservation and Use of Musa Genetic Resources (hereafter referred to as the ‘Global Strategy’) has been expanded in 2016 by Musa genetic resources and breeding experts within the framework of the Global Musa Genetic Resources Network, MusaNet. MusaNet’s mandate is to oversee the further development and monitoring of the implementation of the Global Strategy. The updated Global Strategy aims to provide a clear framework and roadmap to be used by the Musa community for the efficient and effective conservation of the globally important collections of Musa and to strengthen the utilization of the genetic resources toward an increased use of available diversity. It includes recommendations and priorities indicated in several consultation processes following the 2006 Global Musa Strategy and particularly the expertise and key groups represented, including the Regional Research Networks (BAPNET, BARNESA, Innovate Plantain and MusaLAC) and global networks such as ProMusa. The Global Strategy covers numerous topics dealing with Musa genetic resources, with the 12 chapters divided into four main parts: Diversity, Identity, Management and Use. Each chapter contains the sections titled Where we are now, Where do we want to go and How will we get there. For Musa researchers, including taxonomists and breeders, but also end users such as farmers, decisions on the management of banana diversity are often made with limited information. With this in mind, the Global Strategy is a core reference on the taxonomy, characterization, evaluation and genetic improvement of cultivars, leading to actions such as the selection of new and improved cultivars. The use of a more diverse genepool can lead to higher production while at the same time promote ecosystem services such as resilience to pest and disease and the effects of climate change.

Background Myositis-specific autoantibodies (MSA) are increasingly recognized as important diagnostic and prognostic markers in idiopathic inflammatory myopathies (IIM) (polymyositis, dermatomyositis, sporadic inclusion body myositis and necrotizing autoimmune myositis). The prevalence of these MSAs in other systemic autoimmune rheumatic diseases and neuromuscular diseases is unclear. Objectives To compare positivity of MSA in a cohort of IIM patients to positivity in healthy controls and different systemic autoimmune rheumatic diseases (SARD) or chronic inflammatory demyelinating polyneuropathy (CIDP). Methods A line immunoassay (Myositis 12 IgG DOT for BlueDiver) for IgG autoantibodies against Jo-1, PL-7, PL-12, EJ, SRP, Mi-2, MDA-5, TIF1-γ, HMGCR, SSA/Ro52kD, SAE1/2 and NXP-2 antigens was performed in patients with IIM (n=146), healthy controls (blood donors, n=40) and disease controls (n=200). The disease control group consisted of patients with other SARD (rheumatoid arthritis, RA; systemic sclerosis, Ssc; Sjogren9s syndrome, SjS; and systemic lupus erythematosus, SLE) (n=40 for each disease group) and CIDP (n=40). A result >10 arbitrary units was considered significantly positive. Results 50% of 146 patients with IIM tested positive for an MSA (table 1), compared to 3,5% of 200 disease controls (table 1). 1 SSc patient was positive for Jo-1, 1 CIDP patient was positive for PL12, 2 SSc patients were positive for TIF1-gamma, 2 patients (1 SSc and 1 SjS patient) were positive for SAE1/2, and 1 SjS patient was positive for NXP2. The prevalence of SAE1/2 and TIF1-gamma positivity was similar in the IIM and disease control group. No healthy control had a significantly positive MSA. Conclusions MSA positivity in patients with a clinical non-IIM diagnosis (other SARD or CIDP) is infrequent compared to positivity in the IIM group. For TIF1-gamma and SAE1/2 assay performance may need to be optimized. The distribution of subtypes of MSA in this IIM cohort is consistent with data of previous studies.1 References Allenbach Y, Benveniste O. Diagnostic Utility of Autoantibodies in Inflammatory Muscle Diseases. J Neuromuscul Dis. 2015; 2:13–25. Disclosure of Interest None declared

Background Cardiac involvement in systemic sclerosis (SSc) is a frequent complication, but end-stage cardiac failure remains uncommon and represents a poor prognosis. Heart-lung and lung transplant is an established treatment option for SSc-related pulmonary disease. Due to the limited published data, no recommendations exist for cardiac transplant in the context of SSc. Objectives We present our monocentric experience of 3 patients with SSc who underwent cardiac transplant for SSc-related end-stage heart disease (multiple hospitalisations due to failure of medical therapy and life-threatening complications). Results Case 1 is a 59-year-old male with limited cutaneous SSc. Antinuclear antibody (ANA) was negative. He had vascular (digital ulcers) and cardiac (heart failure (left ventricular ejection fraction (LVEF) 20%, NYHA class IV)) involvement, without major gastrointestinal or pulmonary involvement (no interstitial lung disease (ILD) or pulmonary arterial hypertension (PAH: assessed by right heart catheterization (RHC))). He underwent a cardiac transplant at the age of 51, after a disease duration of 6 years. Post-transplantation immunosuppressant therapy consists of tacrolimus and mycophenolic acid, initially associated with methylprednisolon, which is the standard immunosuppression protocol at our institution. Case 2 is a 55-year-old male with limited cutaneous SSc. ANA was positive, 1/320, speckled pattern, but no SSc-related autoantibody has been identified. He had gastrointestinal (upper gastrointestinal tract dysmotility), muscular (myositis) and cardiac (heart failure with secondary cardiac cirrhosis (LVEF 40%, NYHA class III)) involvement, without major pulmonary involvement (no ILD or PAH). He underwent a cardiac transplant at the age of 54, after a disease duration of 7 years. Standard immunosuppressants were initiated. Case 3 is a 50-year-old male with diffuse cutaneous SSc. ANA was negative. He had vascular (digital ulcers), gastrointestinal (upper gastrointestinal tract dysmotility) and cardiac (heart failure with secondary cardiac cirrhosis (LVEF 40%, NYHA class III)) involvement, without major pulmonary involvement (no ILD or PAH). He underwent a cardiac transplant at the age of 49, after a disease duration of 4 years. Standard immunosuppressants were initiated. At present, 1,5 years (case 2 and 3) and 8 years (case 1) after transplant, the donor hearts are still functioning well. No other SSc-related organ manifestations have occurred. Conclusions We present 3 patients with SSc who successfully underwent cardiac transplant for SSc-related end-stage heart disease. None had other major SSc-related organ involvement. This supports the limited published data that cardiac transplant is feasible and can be considered in end-stage SSc-related cardiomyopathy. Disclosure of Interest None declared

The genus Musa is not native to Africa. It evolved in tropical Asia, from southwest India eastward to the island of New Guinea. There is a growing circumstantial evidence which suggests that the East African Highland banana and the tropical lowland plantain were cultivated on the African continent since before 1 AD. It is also probable that ABB cooking and AB and AAB dessert cultivars were brought to the continent from India by Arabian traders from 600 AD, and that these were disseminated throughout East Africa. During the colonial era, the main centres of distribution for banana cultivars were botanical gardens, such as Zomba in Malawi, Entebbe in Uganda and Amani in Tanzania. It appears that the very early introductions of Highland banana and plantain arrived in Africa as a relatively clean material without the conspicuous pests and diseases that affect them in Asia. In contrast, several devastating problems now impact the crop in Africa, including nematodes, the borer weevil and diseases, most notably banana bunchy top, banana streak, Sigatoka leaf spots, Xanthomonas wilt and Fusarium wilt. We (a) provide chronological overviews of the first reports/observations of different Musa pests and pathogens/diseases in Africa, (b) highlight specific examples of when a pest or pathogen/disease was introduced via planting materials and (c) give recent examples of how the pests and pathogens spread to new regions via planting materials. In total, these production constraints threaten banana and plantain production throughout the continent and impact those who can ill afford lost production, the small-holder producer. Our intent in this review is to highlight the significance of these problems and the great importance that infested planting materials have played in their development.

ObjectiveTo identify a core set of preliminary items considered as important for the very early diagnosis of systemic sclerosis (SSc).MethodsA list of items provided by European League Against Rheumatism (EULAR) Scleroderma Trial and Research(EUSTAR) centres were subjected to a Delphi exercise among 110 experts in the field of SSc. In round 1, experts were asked to choose the items they considered as the most important for the very early diagnosis of SSc. In round 2, experts were asked to reconsider the items accepted after the first stage. In round 3, the clinical relevance of selected items and their importance as measures that would lead to an early referral process were rated using appropriateness scores.ResultsPhysicians from 85 EUSTAR centres participated in the study and provided an initial list of 121 items. After three Delphi rounds, the steering committee, with input from external experts, collapsed the 121 items into three domains containing seven items, developed as follows: skin domain (puffy fingers/puffy swollen digits turning into sclerodactily); vascular domain (Raynaud's phenomenon, abnormal capillaroscopy with scleroderma pattern) and laboratory domain (antinuclear, anticentromere and antitopoisomerase-I antibodies). Finally, the whole assembly of EUSTAR centres ratified with a majority vote the results in a final face-to-face meeting.ConclusionThe three Delphi rounds allowed us to identify the items considered by experts as necessary for the very early diagnosis of SSc. The validation of these items to establish diagnostic criteria is currently ongoing in a prospective observational cohort.

In March 2007 the assembly of the Silicon Strip Tracker was completed at the Tracker Integration Facility at CERN. Nearly 15% of the detector was instrumented using cables, fiber optics, power supplies, and electronics intended for the operation at the LHC. A local chiller was used to circulate the coolant for low temperature operation. In order to understand the efficiency and alignment of the strip tracker modules, a cosmic ray trigger was implemented. From March through July 4.5 million triggers were recorded. This period, referred to as the Sector Test, provided practical experience with the operation of the Tracker, especially safety, data acquisition, power, and cooling systems. This paper describes the performance of the strip system during the Sector Test, which consisted of five distinct periods defined by the coolant temperature. Significant emphasis is placed on comparisons between the data and results from Monte Carlo studies., Comment: 40 pages, 39 figures

Dry powder inhalers (DPIs) are increasingly replacing metered dose inhalers in elderly chronic obstructive pulmonary disease (COPD) patients. However, most DPIs are dependent on inspiratory flow, which is compromised by the ageing process itself. Using the in-check dial method, the present study compared peak inspiratory flow (PIF) rates in 26 elderly COPD patients and 14 matched control subjects, at a pre-set resistance level of the Aeroliser, Diskus and Turbuhaler inhalers. It was found that the PIF measured by the in-check method positively correlated with the PIF derived from spirometry, forced vital capacity and maximal inspiratory pressure, while a negative, but significant, correlation was observed with age. PIF derived from spirometry and age were independent variables which determined PIF across the device, whereas the presence or absence of COPD was not related. When comparing elderly COPD patients with matched elderly controls no difference could be found in PIF at the different resistances. However, an important number of patients did not reach the recommended flow rate, especially when using the Turbuhaler (30%). In conclusion, the present study demonstrates that, in elderly patients, the ability to generate sufficient inspiratory flow across a dry powder inhaler is compromised, irrespective of the presence of chronic obstructive pulmonary disease.

Background Pulmonary fibrosis, either idiopathic or secondary to diseases such as systemic sclerosis, is a devastating and life threatening disorder for which effective treatment is still lacking. The bleomycin-induced pulmonary fibrosis model is well-characterized and the most widely used mouse model. The resulting fibrosis is routinely quantified by labor-intensive end-stage histological assessments, requiring many animals and lack the ability to follow-up on disease progression and potential therapeutic effects in the individual animal. At present, imaging tools for the evaluation of lung disease with good temporal and spatial resolution in vivo are limited. Objectives To optimize and evaluate lung MRI protocols to visualize disease onset and progression in the bleomycin-induced model of lung fibrosis. We compared prospectively and retrospectively gated MRI sequences and validated our results with established CT imaging of lung fibrosis and histochemical techniques. Methods Animal Model: Male C57Bl/6 mice were intratracheally instilled with bleomycin (0.05U in 50 µl of PBS) or sham. The mice were scanned with MRI and CT at baseline and weekly until 3 weeks after instillation. After the last imaging time point, mice were sacrificed, ex vivo CT data were acquired and the lungs were isolated for histological analysis and quantification as described before 3. MRI images were acquired at 9.4T (Bruker Biospin, 20 cm) in combination with a 7.5cm quadrature coil, using the following sequences: (1) a respiratory triggered RARE sequence (TR 6000ms TEeff=15.9ms, 50slices of 0.5mm thick), (2) a respiratory triggered ultra short echo (UTE) sequence (FID mode, TR= 20ms, TE=0.4ms, 8 slices, 0.6mm slice thickness) and (3) a retrospectively gated FLASH sequence IntraGate ( TR/TE = 30/1.26 ms, 17 deg flip angle, 5 slices covering the lung, slice thickness 1 mm). For reconstruction, 70% of the respiration and ECG period was used (Paravision 5.1, Bruker)). MRI data were quantified using ImageJ. CT methods: retrospectively gated CT images were acquired on a small animal µCT scanner (SkyScan 1076, Bruker microCT) and quantified. Results The prospectively gated UTE and RARE protocols as well as retrospectively gated IntraGate-FLASH imaging were able to visualize an increase of hyperintense focal spots over time, corresponding to progression of lung fibrosis as corroborated by lung CT images. Quantification of the mean lung signal intensity shows an increase over time, which was confirmed by the decrease in aerated lung volume quantified from the CT data and by histology. UTE, RARE and IntraGate-FLASH images of control animals confirmed the absence of contrast without fibrosis induction. Conclusions The evaluated MRI protocols were all able to non-invasively visualize and quantify lung disease progression. Moreover, the IntraGate-FLASH protocol does not need setup of respiratory triggering for lung imaging, making it an easy to use and efficient alternative to more conventional sequences. Where CT provides poor soft tissue contrast, MRI has the potential to provide contrast differences between vasculature, fibrotic areas and inflammation, without concerns for radiotoxicity when scanning the same animal repeatedly. Disclosure of Interest None Declared

A simple digital-analogue lock-in circuit is described which is used to stabilise the frequency of a laser on the central frequency of a narrow fluorescence line at very low fluorescence intensities (about 102 to 105 detected photons s-1).

2 páginas.-- Póster presentado al 5º European Workshop on Immune-Mediated Inflammatory Diseases celebrado en Sitges (Barcelona, España) del 1 al 3 de Diciembre de 2010.-- et al.