Spondylarthropathies (including psoriatic arthritis): 244. Validity of Colour Doppler and Spectral Doppler Ultrasound of Sacroilicac Joints Againts Physical Examination as Gold Standard

Background: Sacroiliac joints (SJ) involvement is a distinctive and charasteristic feature of Spondyloarthritis (SpA) and x-ray is the test routinely used to make a diagnosis. However, x-ray reveals late structural damage but cannot detect active inflammation. The objective of this study was to assess the validity of Doppler ultrasound in SJ. Methods: Prospective blinded and controlled study of SJ, in which three populations were compared. We studied 106 consecutive cases, who were divided into three groups: a) 53 patients diagnosed with SpA who had inflammatory lumbar and gluteal pain assessed by a rheumatologist; b) 26 patients diagnosed with SpA who didn't have SJ tenderness and had normal physical examination; c) control group of 27 subjects (healthy subjetcs or with mechanical lumbar pain). All patients included that were diagnosed with SpA met almost the European Spondyloarthropathy Study Group (ESSG) classification criteria. Physical examination of the SJ included: sacral sulcus tenderness, iliac gapping, iliac compression, midline sacral thrust test, Gaenslen's test, and Patrick s test were used as gold standard. Both SJ were examined with Doppler ultrasound (General Electric Logiq 9, Wauwatosa WI, USA) fitted with a 9-14 Mhz lineal probe. The ultrasonographer was blinded to clinical data. Doppler in SJ was assessed as positive when both Doppler colour and resistance index (RI) < 0.75 within the SJ area were present. Statistical analysis was performed estimating sensitivity and specificity against gold standard. The Kappa correlation coefficient was used for reliability study. Results: 106 cases (53 female, 55 male; mean age 36 10 years) were studied. There were no statistical differences between groups related to age or sex. Physical examination of SJ was positive in 38 patients (59 sacroiliac joints). US detected Doppler signal within SJ in 37 patients (58 SJ): 33 of them were symptomatic SpA (52 SJ), one of them were asymptomatic SpA (1 SJ) and one was a healthy control (1 SJ). The accuracy of US when compared to clinical data as gold standard at subject level in the overall group was: sensitivity of 68.6% and specificity of 85.7%, positive predictive value of 70.5% and negative predictive value of 84.5%. A positive likelihood ratio of 4.8, a negative likelihood ratio of 0.36 and a kappa coefficient of 0.55 were achieved. Conclusions: Doppler US of SJ seems to be a valid method to detect active SJ inflammation. Disclosure statement: The authors have declared no conflicts of interes

Basic science232. Certolizumab pegol prevents pro-inflammatory alterations in endothelial cell function

Background: Cardiovascular disease is a major comorbidity of rheumatoid arthritis (RA) and a leading cause of death. Chronic systemic inflammation involving tumour necrosis factor alpha (TNF) could contribute to endothelial activation and atherogenesis. A number of anti-TNF therapies are in current use for the treatment of RA, including certolizumab pegol (CZP), (Cimzia ®; UCB, Belgium). Anti-TNF therapy has been associated with reduced clinical cardiovascular disease risk and ameliorated vascular function in RA patients. However, the specific effects of TNF inhibitors on endothelial cell function are largely unknown. Our aim was to investigate the mechanisms underpinning CZP effects on TNF-activated human endothelial cells. Methods: Human aortic endothelial cells (HAoECs) were cultured in vitro and exposed to a) TNF alone, b) TNF plus CZP, or c) neither agent. Microarray analysis was used to examine the transcriptional profile of cells treated for 6 hrs and quantitative polymerase chain reaction (qPCR) analysed gene expression at 1, 3, 6 and 24 hrs. NF-κB localization and IκB degradation were investigated using immunocytochemistry, high content analysis and western blotting. Flow cytometry was conducted to detect microparticle release from HAoECs. Results: Transcriptional profiling revealed that while TNF alone had strong effects on endothelial gene expression, TNF and CZP in combination produced a global gene expression pattern similar to untreated control. The two most highly up-regulated genes in response to TNF treatment were adhesion molecules E-selectin and VCAM-1 (q 0.2 compared to control; p > 0.05 compared to TNF alone). The NF-κB pathway was confirmed as a downstream target of TNF-induced HAoEC activation, via nuclear translocation of NF-κB and degradation of IκB, effects which were abolished by treatment with CZP. In addition, flow cytometry detected an increased production of endothelial microparticles in TNF-activated HAoECs, which was prevented by treatment with CZP. Conclusions: We have found at a cellular level that a clinically available TNF inhibitor, CZP reduces the expression of adhesion molecule expression, and prevents TNF-induced activation of the NF-κB pathway. Furthermore, CZP prevents the production of microparticles by activated endothelial cells. This could be central to the prevention of inflammatory environments underlying these conditions and measurement of microparticles has potential as a novel prognostic marker for future cardiovascular events in this patient group. Disclosure statement: Y.A. received a research grant from UCB. I.B. received a research grant from UCB. S.H. received a research grant from UCB. All other authors have declared no conflicts of interes

Recent advances in ankylosing spondylitis: understanding the disease and management

The term spondyloarthritis refers to a group of immune-mediated diseases characterised by inflammation of the axial skeleton, peripheral joints, and entheses. Ankylosing spondylitis (AS) is the most common and characteristic of these entities and even though it was first described over two centuries ago, the understanding of the underlying disease mechanism remains incomplete. It is known that around 40% of patients with AS have subclinical bowel inflammation, suggesting that the origin of the disease could be in the gut. Also, more genes and new molecules have demonstrated a role in the pathogenesis of AS. In this review, we analyse the latest therapies for spondyloarthritis and the most relevant discoveries over the last three years, together with their implications for different aspects of the disease

P-142 ICSI Semi Automation powered by AI: Gateway for Oocyte Quality Assessment real time based on biomechanics during ICSI

Abstract Study question Embryo selection based on oolemma elasticity and cytoplasm viscosity using semi automated CL-ICSI technology assisted by Artificial Intelligence Summary answer CL-ICSI syringe/technology is capable of converting embryologist visual and feel of each ICSI procedure into a measurable form for objectivity of assessment in real time. What is known already In last four decades, understanding and/or assessing oocyte quality (based on biomechanics) become one of the needs in order to define the embryo formation and selection. initial days of ICSI, embryologist identifies the oocyte pool as fragile, normal or hyper elastic groups based on visual and indentation methods. Further, a variety of advanced techniques were implemented such as ultra fast imaging, atomic force sensing, micro tactile sensors, microfluidics and zone modulus measuring methods. These techniques were implemented on oocyte donation programs, and limited to research, due to the complexity and technical difficulties in implementing. Study design, size, duration From our observations, we would like to introduce our innovative technology (CL-ICSI) that is semi automation of ICSI procedure which is capable of measuring oolemma elasticity and cytoplasmic viscosity. we show the performance using limited number of discarded oocytes as a proof of concept of our technology. We believe that, exploitation of such technology become an asset/gateway for better understanding of oocyte quality and perhaps contribute in embryo selection based on oocyte biomechanics/quality. Participants/materials, setting, methods Current technology (CL-ICSI) is based on ICSI syringes that are calculated/designed to improve objectivity during the ICSI procedure among inter and intra users. CL-ICSI technology comprises of a semi automated syringe equipped with micro controllers that utilises force sensing and pressure mapping techniques to convert oocyte biomechanics data into measurable read outs on display unit. These measurable readouts are finally fed to AI algorithms that calculate the oocyte quality score as an output. Main results and the role of chance CL-ICSI semi automation technology is designed to minimise embryologist subjectivity during the ICSI procedure. Existing syringes works on manual rotatory mechanism which does not provide the start and stop point of the pressure applied on oocyte for puncturing, which genratlly controlled by embryologists skill and experience. Current technology comes with a click button that is designed to be loaded with potential energy which is proportional to the pressure/force required to puncture oolemma subsequently for sperm deposition. In case of classical syringes, post puncturing opposite rotation is implemented to withdraw the negative pressure or cytoplasmic content flow into the needle. Due to the lack of precise positioning the system become subjective. For this reason, we introduced mechanical potential energy build up, where oolemma puncture occurs with a button click and compensation of negative pressure occurs with releasing the button. This mechanism provides objectivity as well requires less than half of the time for oocyte intervention during ICSI over conventional rotatory syringes. More over, this technology works on pneumatics and is adaptable to all existing micro manipulators. Utilisation of such technologies may help improve the objectivity during ICSI and help understand oocyte quality in addition to the visual morphology assessment using AI. Limitations, reasons for caution CL-ICSI technology may be considered as next generation ICSI systems. Application of this technology in clinical setting in collecting the data from multicenters and different ethnicities might provide information that is a step closer to understand the oocyte quality and its role in embryo formation using artificial intelligence. Wider implications of the findings Embryo selection is one of the key event for single embryo transfer for healthy pregnancies. Morphology and morphokinetics data on oocytes and embryos provide valuable information but are limited, if there are no embryos formed. Current technology (CL-ICSI) may provide new avenues for better understanding on embryo prediction and selection. Trial registration number `not applicable` </jats:sec

Prothrombotic gene expression profile in vascular smooth muscle cells of human saphenous vein, but not internal mammary artery

BACKGROUND: In patients with coronary artery disease and reduced ejection fraction, amiodarone reduces mortality by decreasing sudden death. Because the latter may be triggered by coronary artery thrombosis as much as ventricular arrhythmias, amiodarone might interfere with tissue factor (TF) expression and thrombus formation. METHODS AND RESULTS: Clinically relevant plasma concentrations of amiodarone reduced TF activity and impaired carotid artery thrombus formation in a mouse photochemical injury model in vivo. PTT, aPTT, and tail bleeding time were not affected; platelet number was slightly decreased. In human endothelial and vascular smooth muscle cells, amiodarone inhibited tumor necrosis factor (TNF)-alpha and thrombin-induced TF expression as well as surface activity. Amiodarone lacking iodine and the main metabolite of amiodarone, N-monodesethylamiodarone, inhibited TF expression. Amiodarone did not affect mitogen-activated protein kinase activation, TF mRNA expression, and TF protein degradation. Metabolic labeling confirmed that amiodarone inhibited TF protein translation. CONCLUSIONS: Amiodarone impairs thrombus formation in vivo; in line with this, it inhibits TF protein expression and surface activity in human vascular cells. These pleiotropic actions occur within the range of amiodarone concentrations measured in patients, and thus may account at least in part for its beneficial effects in patients with coronary artery disease

Cyclophilin A differentially activates monocytes and endothelial cells Role of purity, activity, and endotoxin contamination in commercial preparations

BACKGROUND: Cyclophilin A (CyPA) is a cytoplasmic protein secreted under inflammatory conditions. Extracellular CyPA is detected in atherosclerotic plaques and has been observed to activate endothelial cells as well as monocytes. METHODS AND RESULTS: Commercially available recombinant CyPA-induced expression of tissue factor (TF) and vascular cell adhesion molecule-1 (VCAM-1) in human aortic endothelial cells (HAEC). However, CyPA from commercial sources contained lipopolysaccharide at concentrations up to 18.9 ng/ml; moreover, it exhibited low purity as determined by protein spectrum analysis and low activity as assessed by peptidyl prolyl cis-trans isomerase (PPIase) assay. An in-house preparation of pure, active, and uncontaminated CyPA failed to induce endothelial TF or VCAM-1 expression; moreover, it was not chemotactic for HAEC. In contrast, such CyPA exhibited potent chemotactic activity on monocytic THP-1 cells, with a maximal effect on migration occurring at a concentration of 5.5 x 10(-9)mol/l. Pretreatment of CyPA with cyclosporine A prevented its effect on THP-1 cell migration; similarly, PPIase-deficient mutant CyPA protein did not induce migration of these cells. In-house prepared CyPA induced the release of Il-6, but not TNF-alpha, from THP-1 cells. CONCLUSIONS: Commercially available CyPA exhibits low purity and activity and may be contaminated by endotoxin. Pure, active, and uncontaminated CyPA does not induce endothelial TF or VCAM-1 expression; instead, it acts as a potent monocyte chemoattractant and induces monocyte Il-6 release, implying a role for extracellular CyPA in the pathogenesis of atherosclerosis via activation of monocytes rather than endothelial cells