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Background The vertebrae of the cervical spine are rather small and it may be difficult to assess if small areas with signal intensity changes represent the bones, joints or entheses, or derive from the surrounding blood vessels. Objectives To investigate if image resolution affects the assessment of inflammatory and structural lesions of the cervical spine. Methods Forty-nine patients with axial spondyloarthritis according to the ASAS criteria started anti-TNF treatment and had “standard” resolution (std-res) and “high” resolution (high-res) MRI sequences of the cervical spine performed at baseline and after 48 weeks. 3 patients had follow-up scan already after 6–24 weeks due to study exclusion. Std-res: STIR sequence: Voxel size 5.0 mm3 (slice thickness 4.0, spatial resolution 1x1.25); T1W sequence: voxel size 4.5 mm3 (slice thickness 4.0, spatial resolution 0.9x1.25). High-res: STIR sequence: Voxel size 3.1 mm3 (slice thickness 3.5, spatial resolution 0.8x1.11); T1W sequence: voxel size 1.4 mm3 (slice thickness 3.0, spatial resolution 0.6x0.76). Images were assessed in known chronology by an experienced axSpA MRI reader (SJP) blinded to clinical data. High-res and std-res were read in random order. MRI lesions of inflammation, fat and new bone formation were defined according to the Canada-Denmark working group [1,2]. Erosions were not assessed. Results Inflammatory lesions: In 9 of 43 patients (21%), inflammatory lesions were detected in the cervical spine at baseline at std-res, while this was detected in 14 of 43 patients (33%) at high-res. Using high-res, as compared to std-res, 6 patients were reclassified from negative to positive for inflammation, 1 patient was reclassified from positive to negative, and 8/28 patients remained classified as positive/negative, p=0.13 by Exact McNemar test. The mean inflammation score was significantly higher at high-res compared to std-res (1.7 (SD 4.5) vs. 0.8 (SD 2.7), p=0.04 by paired t-test). Fat lesions: 11 of 43 patients (26%) had fat lesions in the cervical spine at baseline using std-res, while 10 of 43 patients (23%) had this using high-res. The mean fat score was significantly higher at high-res compared with std-res (1.6 (SD 3.5) vs. 0.8 (SD 1.8), p=0.02 by paired t-test). Bone spurs/ankylosis: 11 of 43 patients (26%) had bone spurs/ankylosis of the cervical spine at baseline at std-res, while 10 of 43 patients (23%) using high-res. The mean new bone formation score was significantly higher at high-res compared with std-res (2.7 (SD 6.1) vs. 1.4 (SD 3.5), p=0.01 by paired t-test). Responsiveness: Standardized response mean for inflammation score at std-res was 0.15, and at high-res 0.14. Structural lesions remained largely unchanged in all patients. Conclusions More patients were classified as having inflammatory lesions in the cervical spine when using high-res MRI, compared to std-res. Likewise, mean scores of inflammatory lesions, fatty lesions and new bone formation were significantly higher compared with std-res. Further studies are needed to investigate the clinical significance of these findings as well as the frequency of these minor lesions in healthy controls. ClinicalTrials.gov: NCT01029847. References Lambert RGW, et al. J Rheumatol 2009;S84:3–17. Ostergaard M, et al. J Rheumatol 2009;S84:18–34. Disclosure of Interest None declared

Objective.Whole-body magnetic resonance imaging (WB-MRI) is a relatively new technique that can enable assessment of the overall inflammatory status of people with arthritis, but standards for image acquisition, definitions of key pathologies, and a quantification system are required. Our aim was to perform a systematic literature review (SLR) and to develop consensus definitions of key pathologies, anatomical locations for assessment, a set of MRI sequences and imaging planes for the different body regions, and a preliminary scoring system for WB-MRI in inflammatory arthritis.Methods.An SLR was initially performed, searching for WB-MRI studies in arthritis, osteoarthritis, spondyloarthritis, or enthesitis. These results were presented to a meeting of the MRI in Arthritis Working Group together with an MR image review. Following this, preliminary standards for WB-MRI in inflammatory arthritides were developed with further iteration at the Working Group meetings at the Outcome Measures in Rheumatology (OMERACT) 2016.Results.The SLR identified 10 relevant original articles (7 cross-sectional and 3 longitudinal, mostly focusing on synovitis and/or enthesitis in spondyloarthritis, 4 with reproducibility data). The Working Group decided on inflammation in peripheral joints and entheses as primary focus areas, and then developed consensus MRI definitions for these pathologies, selected anatomical locations for assessment, agreed on a core set of MRI sequences and imaging planes for the different regions, and proposed a preliminary scoring system. It was decided to test and further develop the system by iterative multireader exercises.Conclusion.These first steps in developing an OMERACT WB-MRI scoring system for use in inflammatory arthritides offer a framework for further testing and refinement.

This case report describes a patient with scleroderma who developed Mycobacterium intracellulare infection, which for more than a year mimicked worsening of her connective tissue disorder. The patient was diagnosed with scleroderma based on puffy fingers that developed into sclerodactyly, abnormal nail fold capillaries, interstitial lung disease, Raynaud’s phenomenon, esophageal dysmotility, and positivity for rheumatoid factor and anti-SSA antibodies. She developed massive inflammatory changes of the cutis, the subcutis, and the muscle fasciae of the right leg, that after several failed attempts of immunosuppressive treatments were found to be caused by Mycobacterium intracellulare. While she was receiving high-dose prednisolone, as worsening of her connective tissue disease was suspected to be the cause of the inflammatory changes, she had Listeria monocytogenes meningitis and was hospitalized for several weeks, but she recovered from this without sequelae. After Mycobacterium intracellulare infection was diagnosed, she was treated with clarithromycin and rifampicin. Her skin manifestations, arthralgias, and fatigue improved considerably, and the wounds of the right leg healed, unfortunately with significant scarring. Immunodeficiency testing was unremarkable. In summary, an infection with Mycobacterium intracellulare was mistaken for an unusually severe progression of scleroderma.

The sparse number of high-resolution human membrane protein structures severely restricts our comprehension of molecular physiology and ability to exploit rational drug design. In the search for a standardized, cheap and easily handled human membrane protein production platform, we thoroughly investigated the capacity of S. cerevisiae to deliver high yields of prime quality human AQPs, focusing on poorly characterized members including some previously shown to be difficult to isolate. Exploiting GFP labeled forms we comprehensively optimized production and purification procedures resulting in satisfactory yields of all nine AQP targets. We applied the obtained knowledge to successfully upscale purification of histidine tagged human AQP10 produced in large bioreactors. Glycosylation analysis revealed that AQP7 and 12 were O-glycosylated, AQP10 was N-glycosylated while the other AQPs were not glycosylated. We furthermore performed functional characterization and found that AQP 2, 6 and 8 allowed flux of water whereas AQP3, 7, 9, 10, 11 and 12 also facilitated a glycerol flux. In conclusion, our S. cerevisiae platform emerges as a powerful tool for isolation of functional, difficult-to-express human membrane proteins suitable for biophysical characterization.

A 38-year-old woman with systemic lupus erythematosus (SLE) presented with low back pain. Following a diagnosis of SLE at the age of 15 years, the patient had experienced facial rash, polyarthritis...

ObjectiveTo validate the Canada-Denmark (CANDEN) MRI scoring system for the spine in axial spondyloarthritis with updated lesion definitions.MethodsLesion definitions in the CANDEN system were updated and illustrated by a consensus set of reference images. Sagittal spine MRIs of 40 patients with axial spondyloarthritis obtained at baseline and at week 52 after initiation of treatment with the tumour necrosis factor inhibitor golimumab were evaluated in unknown chronology by seven readers blinded to all other data.ResultsCANDEN MRI spine inflammation score had very good reliability for status scores (single-measure intraclass correlation coefficient (ICC) of 21 reader pairs median of 0.91 (IQR 0.88–0.92)) and change scores (ICC 0.88 (0.86–0.92)). CANDEN MRI spine fat score had good to very good reliability for status scores (ICC 0.79 (0.75–0.86)) and moderate to good reliability for detecting change (ICC 0.59 (0.46–0.73)). CANDEN MRI spine bone erosion score and CANDEN MRI spine new bone formation score had slight to moderate reliability for status scores (ICC 0.38 (0.32–0.52) and 0.39 (0.27–0.49), respectively).ConclusionThe CANDEN MRI spine scoring system allows a comprehensive evaluation of inflammation, fat, bone erosion and new bone formation of the spine in patients with axial spondyloarthritis. It demonstrated very good reliability for detecting change in inflammation, moderate to good reliability for detecting change in fat, and slight to moderate reliability for detecting bone erosions and new bone formation. Studies with longer follow-up or patients with more advanced spinal involvement may be needed to reliably detect change in bone erosion and new bone formation scores.Trial registration numberNCT02011386.

This case report describes a patient with arthritis of the large joints, bilateral sacroiliitis, and positive anti-SSA and anti-dsDNA antibody, who received sulfasalazine and shortly thereafter became critically ill. He developed toxic epidermal necrolysis, hemolytic anemia, lymphopenia, markedly elevated ferritin, and muscle wasting. A diagnosis of systemic lupus erythematosus was made, and mycophenolate mofetil and systemic glucocorticoids brought this severe disease under control. Toxic epidermal necrolysis-like lesions and hemophagocytic syndrome have been reported as manifestations of systemic lupus erythematosus. This patient possibly had spondyloarthritis or an undifferentiated connective tissue disease at presentation, and we suggest, based on the timing of events, that sulfasalazine may have acted as a trigger of the severe disease manifestations.

Most purinergic receptors activate intracellular Ca2+ signalling, and in epithelia they stimulate transport of major ions. Aim of the present study on pancreatic ducts was to find whether P2 receptors also regulate cellular Ca2+ transport, such as that via the Na+/Ca2+ exchanger (NCX). Since NCX can also be connected with epithelial Ca2+ transport, we also investigated expression of some Ca2+-handling/transporting proteins. Expression analysis revealed that pancreatic ducts of rat and human duct cell line CFPAC-1 (also PANC-1 and Capan-1) express the Na+/Ca2+ exchanger (splice variants NCX1.3 and 1.7), as well as the Ca2+-sensing receptor and the Ca2+-binding protein calbindin-D 9k. These proteins localize to the luminal and lateral membranes of pancreatic duct epithelia. Ca2+ imaging experiments showed that activation of purinergic P2Y and P2X receptors stimulated NCX in human and rat duct cells. Application of ATP to CFPAC-1 monolayers also stimulated Ca2+ transport from the luminal to the basolateral side. Taken together, these results show that pancreatic ducts express a number of Ca2+-handling/transporting proteins and we propose that these together with purinergic receptors play a role in regulation of duct lumen Ca2+ content.

Selecting the appropriate analytical methods for characterizing the assembly and morphology of polymer-based vesicles, or polymersomes are required to reach their full potential in biotechnology. This work presents and compares 17 different techniques for their ability to adequately report size, lamellarity, elastic properties, bilayer surface charge, thickness and polarity of polybutadiene-polyethylene oxide (PB-PEO) based polymersomes. The techniques used in this study are broadly divided into scattering techniques, visualization methods, physical and electromagnetical manipulation and sorting/purification. Of the analytical methods tested, Cryo-transmission electron microscopy and atomic force microscopy (AFM) turned out to be advantageous for polymersomes with smaller diameter than 200 nm, whereas confocal microscopy is ideal for diameters >400 nm. Polymersomes in the intermediate diameter range can be characterized using freeze fracture Cryo-scanning electron microscopy (FF-Cryo-SEM) and nanoparticle tracking analysis (NTA). Small angle X-ray scattering (SAXS) provides reliable data on bilayer thickness and internal structure, Cryo-TEM on multilamellarity. Taken together, these tools are valuable for characterizing polymersomes per se but the comparative overview is also intended to serve as a starting point for selecting methods for characterizing polymersomes with encapsulated compounds or polymersomes with incorporated biomolecules (e.g. membrane proteins).

To explore if the reliability of synovitis assessment by unenhanced MRI is influenced by different MRI field-strengths, coil types and image resolutions in RA patients. Forty-one RA patients and 12 healthy controls underwent hand MRI (wrist and 2nd--5th metacarpophalangeal joints) at 4 different field-strengths (0.23 T/0.6 T/1.5 T/3.0 T) on the same day. Seven protocols using a STIR sequence with different field-strengths, coils (flex coils/dedicated phased-array extremity coils) and resolution were applied and scored blindly for synovitis (OMERACT-RAMRIS method). A 1.5 T post-contrast T1-weighted sequence was used as gold standard reference. Fair-good agreement (ICC=0.38--0.72) between the standard reference and the different STIR protocols (best agreement with extremity coil and small voxel size at 1.5 T). The accuracy for presence/absence of synovitis was very high per person (0.80--1.0), and moderate-high per joint (0.63--0.85), whereas exact agreements on scores were moderate (0.50--0.66). The intrareader agreement (15 patients and 3 controls) on presence/absence of synovitis was very high (0.87--1.0). Unenhanced MRI using STIR sequence is only moderately reliable for assessing hand synovitis in RA, when contrast-enhanced MRI is considered the gold standard reference. Contrast injection, field strength and coil type influence synovitis assessment, and should be considered before performing MRI in clinical trials and practice. • STIR is only moderately reliable for synovitis assessment, compared with post–contrast-T1-w. • Contrast injection, field strength, and coil type influence synovitis assessment. • Contrast injection is recommended for reliable and reproducible hand synovitis assessment.

Optical spectral transmission imaging (OI) measures the blood-specific absorption of light transmitted through tissue.1–5 OI of joints may measure the level of inflammation in joint diseases such as rheumatoid arthritis (RA), where the inflammatory process is associated with angiogenesis and increased blood flow. We compared OI by Hemics Full Hand Proto with colour Doppler (CD) ultrasonography as gold standard reference for inflammation. Sixty-two patients fulfilling the 2010 European League Against Rheumatism (EULAR)/American College of Rheumatology (ACR) criteria for RA, with no significant deformation of hands or wrists and no surgery for the last 3 months, had OI performed of the wrists and all metacarpophalangeal (MCP) and proximal interphalangeal (PIP) joints. Median age was 62 years, 76% were female, median DAS28 was 3.6 and 71% were anti-CCP-positive. Fifty-eight received disease-modifying antirheumatic drugs (synthetic, biological or in combination), five of these in combination with prednisolone. Four patients did not receive antirheumatic treatment. An OI score of …

Background This study investigates the effect of adalimumab (ADA) in patients with axial spondyloarthritis (axSpA). Objectives In an investigator-initiated randomized, placebo-controlled, double-blinded trial (NCT01029847) the efficacy of adalimumab was investigated in patients with axSpA according to the Assessment of Spondyloarthritis International Society (ASAS) criteria.(1,2) Methods 49 patients with axSpA, BASDAI≥4.0 despite treatment with NSAID and clinical indication for TNFα inhibitor treatment were randomized to s.c. placebo (PLA) or ADA 40 mg eow from baseline to week 6. All patients received ADA from weeks 6 to 24. At week 24, patients with a BASDAI reduction >50% or 2.0 were considered responders. MRI of the spine was scored according to SPARCC Spine Inflammation Index by an experienced reader blinded to all other data. Data from the randomized period (week 0–6) were analysed by Chi-square, unpaired t-tests and Mann-Whitney U tests as appropriate. Results Mean (range) age was 38 years (20–61), symptom duration 12 years (0–45) and time since diagnosis 3 years (0–38). 51% were males, 74% HLA-B27 positive, 20% had a history of uveitis, 4% of psoriasis and 6% of inflammatory bowel disease. At week 24, a BASDAI reduction >50% or 2.0 was attained by 32 of 49 (65%) patients (58% and 72% in the PLA and ADA groups, respectively). 61% of patients achieved ASAS 20 response and 61% ASAS 40 response. From baseline to week 6, the SPARCC MRI spine score improved significantly more in the ADA group than in the PLA group, as did clinical variables (Table): Treatment was discontinued in 5 patients due to adverse events during ADA treatment, due to rash (1), unspecified allergic reaction (1), paresthesia and reduced sensibility of 2 fingers (1), back pain worsening (1), and hospitalization due to myalgia and malaise (1). Conclusions Adalimumab significantly improved clinical and MRI measures of disease activity already at week 6 compared to placebo. 2/3 of patients achieved the clinical endpoint at week 24, confirming the clinical efficacy in axSpA. References Maxwell et al. Cochrane Database Syst Rev. 2015;4:CD005468. Rudwaleit et al. Ann Rheum Dis 2009;68:777–83 Disclosure of Interest None declared

Background On MRI, patients with axial spondyloarthritis (axSpA) demonstrate lesions at several anatomical locations. These locations include the vertebral bodies, e.g. anterior and posterior corners and lesions without relation to the corners, and also the posterior segments of the vertebrae, e.g. the costo-transversal joints, costo-vertebral joints, facet joints, spinous/transverse processes, and surrounding soft tissue. The Canada-Denmark (CanDen) definitions of MRI lesions in the spine allows a detailed evaluation of all these anatomical locations for inflammation, fat, erosions and new bone formation.(1–3) Objectives To assess the anatomical location of positive scores by the CanDen MRI scoring system of the spine. Methods 49 patients with axial SpA (ASAS criteria) and clinical indication for tumor necrosis factor alpha (TNFα) inhibitor, including BASDAI >40 mm, had MRI of the spine performed at baseline. 51% were male. Median symptom duration (range) was 12 (0–45) years. Images were scored using the CANDEN MRI scoring system by a reader blinded to clinical data. Results In total, 10 of 49 patients (20%) had involvement of the costo-transversal joints, facet joints, spinous/transverse processes or soft tissue. Conclusions The most frequent lesion in patients with axSpA were inflammation and fat lesions in the rim of the vertebral bodies (the anterior, posterior, antero-lateral and postero-lateral corner lesions) in the thoracic and lumbar spine. 1/5 of the patients had involvement of the posterior segments. References Ostergaard et al. J Rheumatol 2009;36-S84:18–34 Lambert et al. J Rheumatol 2009;36-S84:3–17 Pedersen et al. J Rheumatol 2009;36-S84:35–38 Chiowchanwisawakit et al. J Rheumatol 2009;36-S84:39–47 Disclosure of Interest None declared

Background Different remission criteria are available for patients with rheumatoid arthritis (RA). None of the criteria includes imaging remission, even though studies have shown that many RA patients in sustained clinical remission have synovitis detectable by ultrasound (US), i.e. grey scale synovial hypertrophy and/or Doppler signal in the synovium. Objectives We aimed to investigate the agreement between different clinical remission criteria and remission defined by US in patients with RA receiving biological therapy in routine care. Methods A total of 117 RA patients in sustained remission (i.e. DAS28 ≤2.6 for ≥1 year and no radiographic progression the last year) on biological therapy were recruited. All patients underwent US of 24 joints for grading of grey scale (GS) synovial hypertrophy (0–3) and synovial Colour Doppler activity (CD) (0–3). Imaging remission was defined in two ways: either GS=0 and CD=0 in all 24 joints, or GS≤1 and CD=0 in all 24 joints. Results Baseline characteristics are shown in table 1. At baseline patients received the following disease-modifying antirheumatic drugs (DMARDs): methotrexate (MTX; 72%), sulphasalazine (3%), azathioprine (2%), leflunomide (2%), no DMARDs (21%) and the following treatment with biological therapy: abatacept (2%), adalimumab (28%), certolizumab (3%), etanercept (28%), golimumab(3%), infliximab (27%), tocilizumab (9%). For patients fulfilling DAS28 remission criteria, US defined remission was present in 9% (GS=0 and CD=0 in all 24 joints, termed “very strict” US remission) or 29% (GS ≤1 per joint and CD=0 in all 24 joints, termed “strict” US remission) and 24% had CD>1 in at least 1 joint (table 1). For patients fulfilling other criteria for clinical remission (CDAI, SDAI and ACR/EULAR remission), patients were in very strict US remission in 12%, 12% and 10%, respectively and in strict US remission in 28%, 29% and 20%, respectively. CD activity >1 in at least 1 joint was present in 22%, 25% and 22% of patients, respectively. The anatomical distribution of the GS synovial hypertrophy was 31% in metacarpophalangeal joints, 27% in metatarsophalangeal joints, 19% in wrists, 10% in knees, 7% in elbows and 6% in ankles. Thus, 33% of the total GS synovial hypertrophy was observed in the feet, which are not included in the DAS28 remission criteria. There were no significant differences in US findings (GS or CD) between patients in remission vs patients not in remission, no matter which clinical remission criteria were used. Conclusions Subclinical synovitis was frequently detected by US in RA patients treated with biological drugs in routine care showing poor agreement between the clinical remission criteria and US remission. Further studies are needed to evaluate the benefit of US remission as a goal for treat to target strategies on patient-reported outcomes and radiographic progression. Disclosure of Interest C. H. Brahe: None declared, L. Terslev: None declared, S. Krabbe: None declared, M. Ostergaard Grant/research support from: Abbvie, BMS, Boehringer-Ingelheim, Eli Lilly, Janssen, Merck, Pfizer, Roche, UCB, Celgene, Sanofi, Regeneron, Novartis, T.Jensen, H. Rogind: None declared, H. Jensen: None declared, A. Hansen: None declared, J. Norregaard: None declared, S. Jacobsen: None declared, K. Ellegaard: None declared, V. Fana: None declared, L. Juul: None declared, T. Huynh: None declared, D. Jensen: None declared, N. Manilo: None declared, K. Asmussen Speakers bureau: Novartis, Abbvie, Pfizer, Berlin Chemie, MSD, UCB, P. Frandsen: None declared, S. Pedersen: None declared, N. Krogh: None declared, M. Hetland: None declared

Background The Spondyloarthritis Research Consortium of Canada (SPARCC) Spine Inflammation Index is a well-established MRI scoring system for vertebral body inflammation.(1) It does not take the exact anatomical location of lesions into account. The Canada-Denmark (CanDen) MRI definitions were developed to allow detailed anatomical evaluation of inflammatory and structural lesions of the vertebral bodies as well as the posterior segments in patients with axial spondyloarthritis (axSpA).(2–3) Objectives The aim of this study was to assess the distribution of scores and the responsiveness of this new method, which combines the CanDen MRI definitions into a scoring system, as compared with the SPARCC Spine Inflammation Index in an axSpA randomized controlled trial. Methods For each disco-vertebral unit, inflammatory lesions of vertebral bodies (VB) in the anterior and posterior corners on central sagittal slices were scored as 1 (small) or 2 (large). Non-corner lesions were scored as 2 (small) or 4 (large). Antero-lateral and postero-lateral inflammatory corner lesions in lateral sagittal slices were scored as 1. Inflammatory lesions in the posterior segments (PS) were each scored as 1. Fat (4), erosion and new bone formation were scored in a comparable way (details not shown due to length restrictions). 49 patients with axSpA and indication for TNFα inhibitor were randomized to placebo or adalimumab 40 mg sc. eow for 6 weeks. MRI of the spine was scored according to CanDen definitions and SPARCC by an experienced reader blinded to the clinical data. Responsiveness was assessed by standardized response mean (SRM) and Guyatt9s responsiveness index (GRI). Values ≥0.8 represent a large degree of responsiveness. Results Conclusions A new anatomy-based scoring system for spine inflammatory and structural lesions was described, demonstrating similar responsiveness as the SPARCC method. Further studies are needed to investigate the additional value of assessing posterior segments. References Maksymowych et al. Arthritis Rheum 2005;53:502–9; Lambert et al. J Rheumatol 2009;36-S84:3–17; Ostergaard et al. J Rheumatol 2009;36-S84:18–34; Pedersen et al. Arthritis Res Ther 2013;15:R216 Disclosure of Interest None declared

Pancreas secretes fluid rich in digestive enzymes and bicarbonate. The alkaline secretion is important in buffering of acid chyme entering duodenum and for activation of enzymes. This secretion is formed in pancreatic ducts, and studies to date show that plasma membranes of duct epithelium express H(+)/HCO(3)(-) transporters, which depend on gradients created by the Na(+)/K(+)-ATPase. However, the model cannot fully account for high-bicarbonate concentrations, and other active transporters, i.e. pumps, have not been explored. Here we show that pancreatic ducts express functional gastric and non-gastric H(+)-K(+)-ATPases. We measured intracellular pH and secretion in small ducts isolated from rat pancreas and showed their sensitivity to H(+)-K(+) pump inhibitors and ion substitutions. Gastric and non-gastric H(+)-K(+) pumps were demonstrated on RNA and protein levels, and pumps were localized to the plasma membranes of pancreatic ducts. Quantitative analysis of H(+)/HCO(3)(-) and fluid transport shows that the H(+)-K(+) pumps can contribute to pancreatic secretion in several species. Our results call for revision of the bicarbonate transport physiology in pancreas, and most likely other epithelia. Furthermore, because pancreatic ducts play a central role in several pancreatic diseases, it is of high relevance to understand the role of H(+)-K(+) pumps in pathophysiology.

Most purinergic receptors activate intracellular Ca(2+) signalling, and in epithelia they stimulate transport of major ions. Aim of the present study on pancreatic ducts was to find whether P2 receptors also regulate cellular Ca(2+) transport, such as that via the Na(+)/Ca(2+) exchanger (NCX). Since NCX can also be connected with epithelial Ca(2+) transport, we also investigated expression of some Ca(2+)-handling/transporting proteins. Expression analysis revealed that pancreatic ducts of rat and human duct cell line CFPAC-1 (also PANC-1 and Capan-1) express the Na(+)/Ca(2+) exchanger (splice variants NCX1.3 and 1.7), as well as the Ca(2+)-sensing receptor and the Ca(2+)-binding protein calbindin-D 9k. These proteins localize to the luminal and lateral membranes of pancreatic duct epithelia. Ca(2+) imaging experiments showed that activation of purinergic P2Y and P2X receptors stimulated NCX in human and rat duct cells. Application of ATP to CFPAC-1 monolayers also stimulated Ca(2+) transport from the luminal to the basolateral side. Taken together, these results show that pancreatic ducts express a number of Ca(2+)-handling/transporting proteins and we propose that these together with purinergic receptors play a role in regulation of duct lumen Ca(2+) content.

Purinergic receptors regulate various processes including epithelial transport. There are several studies on P2 receptors in pancreatic ducts of various species, but relatively little is known about these receptors in human tissue. The aim of this study was to identify purinergic receptors in human pancreatic duct cell lines PANC-1 and CFPAC-1. Expression of P2 receptors was examined using RT-PCR and immunocytochemistry. Both cell lines, and also Capan-1 cells, express RNA transcripts for the following receptors: P2Y1, P2Y2, P2Y4, P2Y6, P2Y11-14 and P2X1, P2X2, P2X4, P2X5, P2X6 and P2X7. Using Fura-2 and single-cell imaging we tested effects of various nucleotide analogues on intracellular Ca(2+) signals in PANC-1 and CFPAC-1 cells. The cell lines responded to all nucleotides with the following efficiency: UTP >or= ATP = ATPgammaS > BzATP. ATP, UTP and ATPgammaS elicited oscillatory responses. BzATP, commonly used to stimulate P2X7 receptors, elicited non-oscillatory and transient Ca(2+) responses. Ivermectin, a potentiator of P2X4 receptors, increased Ca(2+) signals evoked by ATP. The single cell Ca(2+) measurements indicated functional expression of P2Y2 and other P2Y receptors, and notably expression of P2X4 and P2X7 receptors. Expression of P2Y2, P2X4 and P2X7 receptors was confirmed by immunocytochemistry. This fingerprint of P2 receptors in human pancreatic duct models forms the basis for studying effect of nucleotides on ion and fluid secretion, as well as on Ca(2+) and tissue homeostasis relevant in physiology and pathophysiology of pancreas.

Background A multi-biomarker disease activity (MBDA) score has been validated as a measure of disease activity in rheumatoid arthritis (RA). Objectives This study aimed to further validate the MBDA score in relation to “imaging inflammation” (MRI bone marrow edema and synovitis, US power Doppler (PD) score and grey-scale synovitis (GSS)) and “imaging damage” (CT/MRI/US erosions, radiographs) in an investigator-initiated trial (HURRAH trial, NCT00696059). Methods Fifty-two RA patients with active disease despite conventional DMARD therapy were included. All started adalimumab treatment at baseline and continued on methotrexate. MRI, CT, radiographs and US were performed at baseline, week (w)26 and w52. Blood tests were drawn at baseline, w2, 6, 12, 26, 39 and 52 for measurement of 12 serum biomarkers (VCAM-1, EGF, VEGF-A, IL-6, TNF-RI, MMP-1, MMP-3, YKL-40, leptin, resistin, SAA, CRP) used to generate a Vectra® DA algorithm score. Associations between MBDA score, DAS28-CRP and imaging inflammation and damage were evaluated using Spearman9s rank correlations (rho). Smallest detectable difference (SDD) was defined as 2 SD of the intrareader difference. Results Median (IQR) MBDA score was 63 (47-74) at baseline, declined to 48 (35-65) already at w2 and remained at roughly this level at all following visits. Change in MBDA score correlated with change in DAS28 from baseline to w26 (rho=0.71, p 10 patients (19%) had progressive joint damage above the SDD by one or more imaging modalities from baseline to w52. Two of these 10 patients were in clinical remission (DAS28 Baseline MBDA score and change/percent change in MBDA score at w2 or w6 generally did not correlate to change in imaging scores at w26 and w52. However, percent change in MBDA score at w6 correlated to change in MRI synovitis score from baseline to w26 (rho=0.44, p=0.009), and percent change in MBDA score at w2 correlated to change in Sharp/van der Heijde total score from baseline to w26 (rho=0.45, p=0.004). In similar analyses with DAS28 no correlations were found at 0.01 significance level. 15 patients achieved clinical remission (DAS28 Conclusions This study further validates the MBDA score as a measure of disease activity in RA, as it correlates with imaging measures of inflammation and joint damage. Disclosure of Interest S. Krabbe: None declared, R. Bolce Shareholder of: Shareholder of Crescendo Bioscience, Employee of: Employee of Crescendo Bioscience, C. Brahe: None declared, U. Dohn: None declared, G. Wu Employee of: Employee of Crescendo Bioscience, B. Ejbjerg: None declared, M. Hetland: None declared, E. Sasso Shareholder of: Shareholder of Crescendo Bioscience, Employee of: Employee of Crescendo Bioscience, D. Chernoff Shareholder of: Shareholder of Crescendo Bioscience, Consultant for: Consultant for Crescendo Bioscience, M. Hansen: None declared, L. Knudsen: None declared, A. Hansen: None declared, O. Madsen: None declared, M. Hasselquist: None declared, J. Moller: None declared, M. Ostergaard: None declared DOI 10.1136/annrheumdis-2014-eular.4460

recognized as important extracellular signaling molecules in many biological systems (1). In epithelia, ATP signaling is thought to regulate fluid transport, and exocrine glands are one of the first epithelia where the effect of ATP on ion channels and Ca signaling was observed (2, 3). Nevertheless, many steps in ATP signaling, also known as purinergic signaling, remain to be clarified. This includes the questions of where and how ATP is released, which purinergic receptors are stimulated and where and how ATP is hydrolyzed by ecto-nucleotidases. In general, our studies aim to understand the role of purinergic signaling in communication between pancreatic acini and ducts. We have established that pancreatic acini release ATP and nucleotidases (4, 5) possibly from zymogen granules and we propose that pancreatic ducts are important sites for purinergic signaling, where the effect on specific receptors is translated into epithelial cell responses. Therefore, our recent studies focused on determining expression and effect of purinergic and adenosine receptors and their effect on Ca and Cl transport.

Objectives With contrast-enhanced MRI as the reference method, to explore to which extent synovitis in wrists and metacarpophalangeal (MCP) joints of patients with rheumatoid arthritis (RA) can be reliably assessed by unenhanced MRI. Furthermore, we explored the influence of different MRI field strengths, coil types and image resolutions. Methods 41 RA patients and 12 healthy controls underwent 7 MRI examinations (Coronal short tau inversion recovery (STIR) and T1-weighted (T1w) sequences) on 4 MRI units (0.23 T, 0.6 T, 1.5 T, and 3.0 T, with different coils, see table for details) within 24 hours. Additional post-contrast T1w sequence was performed at the 1.5T unit (gold standard). Images were scored according to OMERACT RA MRI score (RAMRIS) (1) by an experienced reader blinded to participant’s details and MRI field strength. Gold standard images were scored on a different session. Intrareader reliability was evaluated on a subset of 18 patients and controls. Results Fair to good correlation (ICC=0.38-0.72) was seen between different STIR protocols and the gold standard reference (Table 1). The highest correlation was found using the 1.5T unit with the smaller voxel size and extremity coil. Sensitivity for the presence or absence of synovitis was moderate-high in all units (per patient: 0.8-1.0, per joint: 0.63-0.85) whereas the exact agreement on scores was poorer (0.5-0.66). Conclusions Unenhanced MRI, using STIR sequences, is only moderately reliable for assessing synovitis in RA MCP and wrist joints, when contrast-enhanced MRI is considered the gold standard reference. 0.6 T and 1.5 MRI performed best. References Ostergaard M. et al. J Rheumatol 2003;30:1385-6. Disclosure of Interest None Declared

OBJECTIVE The aim of this study was to examine the influence of different MRI unit field strengths, coil types and image resolutions on the OMERACT RA MRI scoring system (RAMRIS) of bone marrow oedema (BME) and image quality. METHODS Forty-one patients and 12 healthy controls participated in this cross-sectional study. Coronal short tau inversion recovery (STIR) and T1-weighted sequences were obtained at 0.23, 0.6, 1.5 and 3T using flex coils (Flex). Additional STIR sequences were obtained with phased array extremity coils (Extr) (at 0.6 and 1.5T) and higher resolution (at 1.5T). In otal, 338 STIR image sets were anonymized and scored according to RAMRIS and parameters of image quality were measured. RESULTS The BME sum scores were similar overall when comparing the different MRI units, coil types and voxel sizes, yet significantly higher at the higher resolution of 1.5T Extr compared with 0.23T Flex (P = 0.004), 0.6T Flex (P = 0.03), 1.5T Flex (P = 0.05) and 3T Flex (P = 0.001). Mean differences were relatively minor (0-3.5). Intrareader reliability of BME scores was high [intraclass correlation coefficient ≥ 0.90 for all except 0.23T (0.81) and percentage exact agreement 81-88%]. The smallest detectable difference was better at 0.6, 1.5 and 3T (9-29% of maximum value) than at 0.23T (40%). Image quality was lowest at 0.23T. CONCLUSION No major, consistent differences were found between BME scores using STIR sequences obtained at different field strengths, coil types and image resolutions, suggesting that these are equally suited for assessment of BME in RA. However, parameters of image quality and intrareader reliability (favouring 0.6, 1.5 and 3T) should be considered when selecting the MRI acquisition strategy.