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A retrospective study found more harm than benefit from treating elevated blood pressure in hospitalized noncardiac patients.

Virtual Reality (VR) is progressively adopted at different stages of design and product development. Consequently, evolving interaction requirements in engineering design and development for VR are essential for technology adoption. One of these requirements is real-time positional tracking. This paper aims to present an experimental design of a new real-time positional tracking device (tracker), that is more compact than the existing solution, while addressing factors such as wearability and connectivity. We compare the simulation of the proposed device and the existing solution, discuss the results, and the limitations. The new experimental shape of the device is tailored towards research, allowing the engineering designer to take advantage of a new tracker alternative in new ways, and opens the door to new VR applications in research and product development.

Additive Manufacturing is enabling the casting of complex geometries directly from digital design data, including 3D-scanned and reverse-engineered structures and even functionally graded lattices. By ink jetting binder into a bed of sand layer-by-layer, dimensionally precise sand molds and cores can be printed to serve as soft tooling for sand casting. However, the related increase in geometry complexity can lead to challenges in ensuring casting quality and yield. One recently explored remedy is to introduce sensors (the Internet of Things) to enable the collection of a diversity of data at difficult-to-access locations in molds in order to measure temperature, pressure, moisture, and core shift. This effort has explored measuring barometric pressure at strategic locations to evaluate the ventilation design of internal cores. Optimized and measurable ventilation can be leveraged to improve the quality of castings by reducing porosity and improving surface finish. By measuring the pressures that accumulate within cores due to binder decomposition, new ventilation designs and strategies—enabled with complex, 3D printed fluidic channels—can be explored. In this work, two castings with different metal temperatures were poured and internal pressures were measured and compared to simulations demonstrating that wireless disposable sensors can be used to measure pressure and that the measured pressure correlated with venting strategies now possible with 3D printed sand cores.

3D sand printing is an emerging technology that is enabling new possibilities in metal casting with respect to part complexity, casting design, and rapid mold production. The 3D printing technology, also known as additive manufacturing, is a binder jetting process which involves selectively depositing a furan-based binder into a sand bed one thin layer at a time. Over the course of the process, layers are subsequently added until the entire part has been fabricated. Although the use of 3D-printed sand molds in the foundry industry is growing, significant hesitation to widespread implementation remains. In this work, an investigation was conducted to determine the influence of machine settings on the physical characteristics of 3D-printed sand. Two factorial matrices were constructed to directly measure the significance of six settings that change the resin content and compaction characteristics of the bonded sand. Factors include: X-resolution, printhead voltage, layer thickness, and the frequency, speed, and angle of the recoater blade. Responses include: density, permeability, strength, scratch hardness, loss on ignition, and print resolution. Several relationships are reported between machine settings and physical properties of the product. These results will help inform mold manufacturing as the foundry industry continues to adopt additive technologies.

A recent Australian study demonstrated a significant reduction in A-fib recurrence and burden among regular drinkers who abstained from alcohol.

While they have long been a subject of academic study, social virtual reality (SVR) systems are now attracting increasingly large audiences on current consumer virtual reality systems. The design space of SVR systems is very large, and relatively little is known about how these systems should be constructed in order to be usable and efficient. In this paper we present Ubiq, a toolkit that focuses on facilitating the construction of SVR systems. We argue for the design strategy of Ubiq and its scope. Ubiq is built on the Unity platform. It provides core functionality of many SVR systems such as connection management, voice, avatars, etc. However, its design remains easy to extend. We demonstrate examples built on Ubiq and how it has been successfully used in classroom teaching. Ubiq is open source (Apache License) and thus enables several use cases that commercial systems cannot.

BackgroundAnti-neutrophil cytoplasmic autoantibodies specific for myeloperoxidase (MPO-ANCA) are found in 10-70% of the patients with eosinophilic granulomatosis with polyangiitis (EGPA) depending on disease activity, methodological aspects and cohort examined [1-3]. Recently, a higher prevalence of anti-pentraxin 3 (PTX3)-ANCA has been reported in EGPA compared to granulomatosis with polyangiitis (GPA) and microscopic polyangiitis (MPA) [4].ObjectivesTo investigate the spectrum of ANCA specificities in a multicenter cohort of patients with EGPA and identify novel ANCA antigens.MethodsWe conducted a retrospective analysis of 73 patients with EGPA treated between 2015 and 2020 in 3 tertiary referral centers. In addition to in-house ANCA testing with indirect immunofluorescence (IFT) on fixed human granulocytes and antigen-specific enzyme-linked immunosorbent assays (ELISA), ANCA specificities were determined using a cell-based assay (CBA; Euroimmun, Lübeck, Germany). Diagnosis was based on Chapel Hill consensus conference definitions, ACR- and MIRRA-criteria for EGPA. Patient characteristics and clinical manifestations were evaluated and compared based on ANCA status. Fisher`s exact test was employed for comparison of patient groups.ResultsANCA findings are summarized in Table 1. MPO- and proteinase 3 (PR3)-ANCA positive patients (13.7%) had a higher prevalence of peripheral neuropathy (70% vs. 44.4%, p = 0.0003) and glomerulonephritis (20% vs. 14.3%, not significant). MPO- and PR3-ANCA-negative patients (86.3%) had a higher prevalence of heart (10% vs. 46%, p vs. 14.3%, p vs. 22.2%, p = 0.0327) involvement. PTX3-ANCA were associated with a higher prevalence of ear-nose-throat (ENT) (100% vs. 85.3%, p vs. 89.7%, p = 0.0015), gastrointestinal involvement (60% vs. 17.6%, p vs. 48.5%, p vs. 16.2%, p vs. 13.2%, p = 0.0002) occurred less frequently in PTX3-ANCA positive patients. The 2 olfactomedin 4 (OLM4)-ANCA positive patients presented with ENT, lung and kidney involvement, and polyneuropathy, respectively.Table 1.ANCA in EGPA cohort (n = 73). BPI = bactericidal permeability-increasing protein.IFT / ELISANo. of patients (%)P-ANCA11 (15.1)C-ANCA5 (6.8)MPO-ANCA8 (10.9)PR3-ANCA2 (2.7)BPI-ANCA1 (1.4)PTX3-ANCA5 (6.8)OLM4-ANCA2 (2.7)ConclusionWe report on the detection of PTX3-, BPI- and OLM4-ANCA in addition to MPO- and PR3-ANCA in EGPA. OLM4-ANCA has been reported in 2 patients with non-vasculitic inflammatory symptoms previously [5]. Herein, detection of OLM4-ANCA in EGPA is reported for the first time. Our study shows that the presence of ANCA with various specificities other than MPO and PR3 contribute to a higher prevalence of ANCA in EGPA. Moreover, clinical manifestations differ between ANCA-negative EGPA and ANCA-positive EGPA, and between patients with different ANCA-specificities.References[1]Schönermarck U, et al. Prevalence and spectrum of rheumatic diseases associated with proteinase 3-antineutrophil cytoplasmic antibodies (ANCA) and myeloperoxidase-ANCA. Rheumatology 2001;40:178-84.[2]Bremer P, et al. Getting rid of MPO-ANCA: a matter of disease subtype. Rheumatology 2013:752-4.[3]Comarmond C, et al. Eosinophilic granulomatosis with polyangiitis (Churg-Strauss): clinical characteristics and long-term followup of the 383 patients enrolled in the French Vasculitis Study Group cohort. Arthritis Rheum 2013;65:270-81.[4]Padoan R, et al. IgG anti-Pentraxin 3 antibodies are a novel biomarker of ANCA-associated vasculitis and better identify patients with eosinophilic granulomatosis with polyangiitis. J Autoimmun 2021;124:102725.[5]Amirbeagi F, et al. Olfactomedin-4 autoantibodies give unusual c-ANCA staining patterns with reactivity to a subpopulation of neutrophils. J Leukoc Biol 2015;97:181-9.Disclosure of InterestsNone declared

Idiopathic pulmonary fibrosis (IPF) pathogenesis has been postulated to involve a variety of mechanisms associated with the aging process, including loss of protein homeostasis (proteostasis). Heat shock proteins are cellular chaperones that serve a number of vital maintenance and repair functions, including the regulation of proteostasis. Previously published data have implicated heat shock protein 70 (Hsp70) in the development of pulmonary fibrosis in animal models. We sought to identify alterations in Hsp70 expression in IPF lung. Hsp70 mRNA and protein were decreased in primary fibroblasts cultured from IPF versus normal donor lung tissue. In addition to cultured fibroblasts, Hsp70 expression was decreased in intact IPF lung, a stressed environment in which upregulation of protective heat shock proteins would be anticipated. In support of a mechanistic association between decreased Hsp70 and fibrosis, cultured primary lung fibroblasts deficient in Hsp70 secreted increased extracellular matrix proteins. Treatment of primary normal human lung fibroblasts in vitro with either of the profibrotic molecules IGFBP5 (insulin-like growth factor-binding protein 5) or transforming growth factor-β1 downregulated Hsp70, suggesting Hsp70 is a downstream target in the fibrotic cascade. Hsp70-knockout mice subjected to an inhalational bleomycin model of pulmonary fibrosis demonstrated accelerated fibrosis versus wild-type control animals. We therefore conclude that reduced Hsp70 protein contributes to fibrosis and that interventions aimed at restoring normal expression of Hsp70 represent a novel therapeutic strategy for pulmonary fibrosis.

In this work, real-time in-process monitoring of core motion in metal castings is demonstrated through the use of two emerging technologies. 3D sand printing (3DSP) is a binder jetting additive manufacturing process that is quickly manifesting itself as a technological disrupter in the metal casting industry. Based on its direct digital manufacturing principle, 3DSP enables complex mold and core design freedom that has been previously unavailable to foundry engineers. In addition, the miniaturization and affordability of electronics and sensing equipment is rapidly accelerating. Here, these two shifting paradigms are leveraged together. An experimental casting and mold were designed in this research to demonstrate and evaluate wireless sensing of core shifts. With the use of 3D sand printing, precisely sized and located pockets were manufactured inside of cores. Miniature wireless Bluetooth sensors capable of measuring acceleration and rotation were then embedded inside the cores. From these, high fidelity data were captured wirelessly from the sensors during the casting process. With strategically designed core prints designed to allow varying levels of core motion, it is shown that core shifts can be measured and discriminated during casting in real time.

For many years, researchers in the foundry industry and universities have been developing testing protocols to evaluate the physical, performance and emission characteristics of prepared green sand during pouring, cooling and shakeout. This paper will review the testing protocol and methods utilized in the collection and evaluation of these characteristics that is the result of the entire metal casting process. An important criterion also is the quality of the green sand mold and casting produced (defect free) when the emission characteristics are reduced. This paper will also include the laser imaging evaluation of the castings that is the result of the green sand foundry emissions.

Playing virtual reality (VR) games while seated can be challenging because many games are designed for a mobile, standing player. Sitting down not only limits mobility but also the reach of the player. This is particularly problematic if the player does not have a choice in whether they play seated or standing because it then becomes an accessibility issue. We have taken the concept of co-piloting (controls shared across multiple input devices and players) and applied it to VR with the aim of extending the reach of the VR player. To demonstrate the feasibility of the concept, a software prototype was developed that allowed a second, remote, player to take over the VR player’s controllers and help them reach out. Because the prototype manipulates the SteamVR middleware and not the game, it can be used with any application that runs on SteamVR. A small formative study was conducted to inform further research and proof the concept.

BackgroundCytotoxic Natural Killer (NK) cells are an important target of new drugs entering the clinics, including checkpoint inhibitors and cell-depleting therapeutic antibodies. Still, basic blood NK cell parameters are poorly defined in healthy adults and in chronic inflammatory diseases like ANCA-associated vasculitis (AAV) which may alter the distribution of lymphocytes. The aims of this study were 1) to establish reference values of NK cell counts and percentages in healthy adults; 2) to describe these parameters in AAV; and 3) to investigate whether NK cell counts and percentages may be used as activity biomarker in the care of AAV patients, as suggested by a preceding study.MethodsCD3-(CD56 or 16)+ NK cell counts and percentages were determined in 120 healthy adults. Lymphocyte subset data from two German vasculitis centers were retrospectively analyzed (in total 407 measurements, including 201/49/157 measurements from 64/16/39 patients with granulomatosis with polyangiitis (GPA), microscopic polyangiitis (MPA) and eosinophilic granulomatosis with polyangiitis (EGPA), respectively).ResultsCD3-(CD56 or 16)+ NK cell counts and percentages in healthy adults were highly variable, not Gaussian distributed and independent of age and sex. NK cell percentages ranged from 1.9 to 37.9% of lymphocytes, and were significantly more dispersed in AAV (0.3 to 57.6%). We further found that NK cell counts and percentages were different between AAV entities. However, during active disease, NK cell counts were consistently low in each AAV entity compared to healthy donors. NK cells were especially low in inactive EGPA. In 18% of EGPA patients we observed percentages of 1% or below which may be interpreted as temporary NK cell deficiency. Findings on differences between active and inactive GPA were discrepant between vasculitis centers.ConclusionsNK cell counts and percentages in blood are highly variable. This variability is further enhanced in systemic inflammatory diseases, and includes patients with temporary NK cell deficiency, in particular in EGPA. NK cell counts and percentages can presently not be recommended as biomarker in clinical care of AAV patients.

BackgroundB differentiation in the bone marrow (BM) is impaired in patients carrying mutation in the IKFZ1 gene, coding for Ikaros a zinc-finger transcription factor. High Ikaros expression is on the contrary associated with systemic lupus erythematosus[1] and multiple myeloma[2]. Targeted treatment with iberdomide, a cereblon modulator which enhances degradation of Ikaros and Aiolos, is under clinical investigation in multiple myeloma patients and systemic lupus erythematosus. However, consequences of the treatment on human early B cell development remain elusive. Immature B cells develop in the BM from hematopoietic stem cells. An intricate network of transcription factors regulates the maturation process. Ikaros and Aiolos regulate gene expression during B cell development. As reported in mice, Ikaros is essential for the commitment to the lymphoid lineage and later, together with Aiolos, ensures the transition from pre-BII large to pre-BII small cells.ObjectivesInvestigate the effect of iberdomide (CC-220) on human early B cell development simulated in vitro.MethodsWe tested the impact of iberdomide on short term culture of BM-derived lymphocytes and in a unique in vitro modeling of early B cell development starting from cord blood (CB)- CD34+ progenitors [3, 4]. We used multi-dimensional spectra flow cytometry (17-color pan-el) to dissect early B cell subpopulations.ResultsIberdomide treatment led to enhanced degradation of Ikaros and Aiolos in both BM- and CB-derived cultures. Addition of iberdomide early (day 7) to the CB-derived culture impaired the specification to the lymphoid lineage and later also the commitment to the B cell lineage. These observations were confirmed by reduced E2A and PAX5 gene expression, respectively. Treatment with iberdomide on B cell precursors (pro- and pre-B cells, day 28 of culture) on one side it enhanced the proliferation of early progenitors resulting in increased amount of CD10+CD38+ lymphoid-committed cells. On the other side, it resulted in a accumulation of pre-B cells and inefficient development of immature B cells.ConclusionIberdomide impairs the commitment to the lymphoid lineage by enhancing Ikaros’ degrada-tion. When targeting already committed B cells, iberdomide treatment undermines the transition of pre-BII large to pre-BII small cells due to increased Aiolos’ degradation, conse-quently impairing the development of immature B cells. Our data can instruct immunologi-cal monitoring of patients treated with iberdomide, and provide insights in the mechanisms of therapeutic efficacy.References[1]Rivellese, F., et al., Effects of targeting the transcription factors Ikaros and Aiolos on B cell activation and differentiation in systemic lupus erythematosus. Lupus Sci Med, 2021. 8(1).[2]Thakurta, A., et al., Developing next generation immunomodulatory drugs and their combinations in multiple myeloma. Oncotarget, 2021. 12(15): p. 1555-1563.[3]Kraus, H., et al., A Feeder-Free Differentiation System Identifies Autonomously Proliferating B Cell Precursors in Human Bone Marrow. The Journal of Immunology, 2014. 192(3): p. 1044-1054.[4]Troilo, A., et al., Nonpermissive bone marrow environment impairs early B-cell development in common variable immunodeficiency. Blood, 2020. 135(17): p. 1452-1457.Disclosure of InterestsIga Janowska: None declared, Jakov Korzhenevich: None declared, Julian Staniek: None declared, Raquel Lorenzetti: None declared, Lukas Konstantinidis: None declared, Miriam Erlacher: None declared, Peter Schafer Employee of: BMS, Reinhard Voll: None declared, Jens Thiel Grant/research support from: BMS (former Cellgene), Nils Venhoff: None declared, Marta Rizzi Grant/research support from: BMS (former Cellgene)

BackgroundRespiratory tract infections (RTIs) are the most common infections in patients with rheumatic diseases under immunosuppressive treatment. RTIs may cause significant morbidity with reduced quality of life (QOL), increased healthcare costs and may lead to interruption of DMARD therapy. However, to date only limited data on infection risk in spondyloarthritis (SpA) patients are available.ObjectivesTo assess the occurrence of respiratory tract infections in a real-world SpA cohort and determine associated factors.MethodsQuestionnaire-based screening and retrospective medical chart analysis of a monocentric cohort of 314 SpA patients comprising 168 psoriatic arthritis (PsA) and 146 axial spondyloarthritis (axSpA) patients.ResultsOut of 314 SpA patients, 89% had a history of upper respiratory tract infections (URTI) and 31.1% of lower respiratory tract infections (LRTI) within the last two years (Table 1). In a linear regression model LRTIs were associated with glucocorticoid (GC) therapy (p=0.015), CRP level (p=0.018), previous history of severe respiratory infections (p=0.007) as well as absence of HLA B27 (p=0.024). In general, patients with LRTIs were significantly older (p=0.007), had increased functional impairment (pTable 1.Patient characteristics and infectionsaxSpA (n=146)PsA (n=168)Total (n=314)Age, years, mean (SD)49.6 (14.2)57.4 (12.4)53.8 (13.9)Male / Female, %56.8 / 43.253.0 / 47.054.8 / 45.2BMI, kg/m2, mean (SD)27.1 (7.2)27.4 (5.4)27.3 (6.2)Smokers, n (%)41 (31.8)27 (16.3)68 (23.1)HLA B27, n (%) (n=230)97 (71.9)23 (24)120 (52.2)Therapy:n (%) csDMARD29 (19.6)84 (50.6)113 (36.2) bDMARD109 (75.2)102 (60.7)211 (67.4) Glucocorticoids14 (9.9)15 (8.9)29 (9.4)Hypogammaglobulinemia (IgG5 (3.5)6 (3.6)11 (3.6)Polyclonal IgA (>4g/l) elevation, n (%)15 (10.9)29 (17.8)44 (14.7)URTI: n (%)114 (88.4)148 (90.2)262 (89.4)Rhinitis, %87.187.587.3Laryngitis/pharyngitis, %3736.936.9Sinusitis, %40.530.234.7Otitis media, %14.36.810.1LRTI: n (%)39 (30.5)52 (32.1)91 (31.1)Bronchitis, %28.030.629.6Pneumonia, %3.93.13.4Pleuritis, %2.41.21.7There were no significant differences between PsA and axSpA regarding frequency of URTI or LRTI, though PsA patients had tendentially more overall RTIs. Biological therapy did not lead to a significantly increased occurrence of infections, but was associated with increased antibiotic therapy (p=0.039). Patients with a history of pneumonia had received anti-IL17 therapy more frequently (p=0.002), while there was no significant association with anti-TNF therapy (p=0.156). Patients on GC had a relative risk for LRTIs of 2.04.Hypogammaglobulinemia was rare in SpA patients (3.6%) despite continuous immunosuppressive treatment, occurred with equal frequency in axSpa and PsA patients and was associated with pneumonia (p=0.007) and increased antibiotic use (p=0.016). Polyclonal IgA elevation was observed in 14.7% of patients (mean 4.98g/l) and was associated with fewer episodes of rhinitis (p=0.027), whereas LRTIs and antibiotic use did not differ significantly.ConclusionThis study quantifies the incidence and effects of RTIs in a real-world SpA cohort. While infections constitute significant adverse events of biologicals, and URTI were common, severe respiratory tract infections were rare. Differences in infection risk between SpA and PsA need to be studied more closely.Disclosure of InterestsNatalie Frede Grant/research support from: Novartis study grant, Eva Rieger: None declared, Raquel Lorenzetti Grant/research support from: Novartis study grant, Ana Venhoff: None declared, Anna-Maria Kanne: None declared, Marcus von Deimling: None declared, Nora Bartholomä: None declared, Jens Thiel Speakers bureau: Novartis, AbbVie, Pfizer, BMS, UCB, Consultant of: Novartis, AbbVie, Pfizer, BMS, UCB, Grant/research support from: BMS, Novartis study grants, Reinhard Voll Speakers bureau: Novartis, AbbVie, Pfizer, BMS, UCB, Consultant of: Novartis, AbbVie, Pfizer, BMS, UCB, Lilly, Grant/research support from: Novartis study grant, Nils Venhoff Speakers bureau: Novartis, AbbVie, Pfizer, BMS, UCB, Consultant of: Novartis, AbbVie, Grant/research support from: Novartis study grant

BackgroundPrimary Sjögren Syndrome (PSS) is an autoimmune disorder with a diverse spectrum of clinical manifestations ranging from sicca symptoms to severe systemic organ involvement. Little is known about the symptoms at the onset of PSS, as these are often ignored by both, patients and physicians leading to a substantial delay of diagnosis.ObjectivesThe aim of this study was to investigate patients’ recollection of the first symptoms before diagnosis of PSS in qualitative interviews. The second aim was to verify and quantify these aspects in a representative cohort.MethodsAll PSS patients fulfilled the EULAR/ACR 2016 classification criteria. In the first part of the study, consecutive PSS patients were recruited for individual, semi-structured interviews. A discussion guide with five open-ended questions was developed to explore patients’ experiences on the onset of PSS. All interviews were audio-recorded and transcribed verbatim, and an inductive thematic data analysis was performed using MAXQDA software (VERBI, Berlin, Germany).In the second part, the identified aspects of the qualitative analysis were grouped to a checklist with ten items. Patients were asked to complete the checklist before their routine clinical assessment.ResultsOne-hundred and thirty-four patients participated in the study. The qualitative part was completed by 31 PSS patients; 90.3% (n=28) were female and patients had a mean disease duration of 6.9 years (±5.7(SD)) and a mean age of 58.1 years (±12.6).Four different major aspects emerged of how patients experienced the beginning and first symptoms of PSS: (1) sicca symptoms started after initial swelling of parotis and/or lymph nodes (2) “Classic” PSS symptoms (fatigue, pain, dryness): patients reported wandering joint pain before diagnosis with a long time apart from first symptoms until diagnosis. Patients described joint pain, chronic malaise, and fatigue over months. (3) Hormonal changes (e.g. after birth, hysterectomy) or infections before the onset of PSS symptoms. (4) Slowly progressing discomfort due to sicca: patients reported a slow progression of symptoms with no initial recognition of sicca discomfort. In these patients recurrent dental problems and loss of teeth in the years prior to diagnosis was common.In the second part of the study, the four themes were verified in an independent cohort of 103 PSS patients. Patients were 59.9 (±13.7) years old and six patients were male. The main symptom before diagnosis was dryness (n=77, 74.8%) with wandering joint pain (n=51, 49.5%) and fatigue (n=47, 45.6%). In 38.8% (n=40), patients reported a swelling/inflammation of the parotid gland at the onset of disease.ConclusionWe identified four themes describing the initial symptoms of PSS. Raising awareness of these symptoms among physicians and among the general public may allow earlier diagnosis of PSS.Disclosure of InterestsNone declared

BackgroundCD19 is a membrane glycoprotein interacting with different surface molecules like the B cell receptor (BCR) and is crucial for antigen-independent development as well as immunoglobulin-induced activation of B cells.[1] Alterations in this signalling pathway can incline autoantibody production and systemic autoimmunity in humans. Rituximab (RTX), a CD20 antagonist appears to be an effective candidate in the treatment of different autoimmune diseases that are partly driven by autoreactive B cells, such as systemic sclerosis (SSc).[2] It has been speculated that RTX might work not only by depleting B cells but also to down regulate activation markers, such as CD19.ObjectivesIn-depth analysis of CD19 abundancy and activation on B cells in SSc patients with and without RTX treatmentMethodsPeripheral blood samples from 41 patients suffering from SSc (median ± standard deviation SD, age: 54.3 ± 10.6 years, female ratio: 0.8) and 45 age- and sex-matched healthy controls (HC) (age: 51.0 ± 13.9 years, female ratio: 0.8) were drawn and PBMCs were isolated on-site. We performed flow cytometry analysis on a standardized BD LSRFortessa platform to identify B cell (CD19+CD20+) subpopulations. The geometric mean fluorescence intensity (gMFI) for CD19 in all B cell subtypes was extracted from the data set and used for further statistical analysis. Additionally, a quantitative flow cytometric bead-based assay (QuantiBRITE PE kit from Becton Dickinson) was used for the estimation of CD19 antibodies bound per cell.Results3 out of 41 SSc patients were in high disease activity at the time of blood drawal. 23 SSc patients were under RTX therapy of whom 5 patients still displayed measurable B cells frequencies. Naïve B cells made up the most abundant B cell population in SSc patients. Thus, the frequency of IgM+/IgD+/CD27- B cells was 67.9% ±13.2 (mean ±SD), followed by class-switched memory B cells (IgM-/IgD-/CD27+, 10.5 ± 4.9), non-switched memory B cells (IgM+/IgD+/CD27+, 4.0 ± 3.6) and plasmablasts (0.3 ± 4.4). Pairwise Wilcoxon Tests (Bonferroni-corrected for multiple testing) showed significant differences (p < 0.001) between frequencies of naïve B cells and all other cell types. In contrast, naïve B cells displayed the second lowest CD19 gMFI levels (7601.0 ± 1912.0) in the dataset. Non-switched memory B cells in SSc patients showed the highest CD19 gMFI (10620.0 ± 15689.8), followed by class-switched (9388 ± 3048.6). As expected, Plasmablasts displayed the lowest CD19 gMFI levels (4799.0 ± 4185.7). The decrease in CD19 gMFI was again highly significant. This trajectory in decreasing CD19 gMFI was found in both HCs and SSc patients. We saw a significant reduction in percentages of non-switch B cells and class-switched B cells in SSc patients compared to HCs (4.0 ± 3.6 vs 6.5 ± 4.2, p = 0.029, 10.5 ± 4.9 vs 13.2 ± 7.2 p = 0.04) but an increase in CD19 gMFI in non-switched B cells (HC: 9204.5 ± 2116.8, p = 0.05). Interestingly, SSc patients under RTX treatment had significantly lower class-switched memory B cell frequencies compared to HCs (6.4 ± 4.2 vs. 13.2 ± 7.2, p = 0.015). However, RTX did not affect CD19 gMFI or bound CD19 in SSc.ConclusionRTX treatment in SSc is not associated with downregulation of the co-stimulation marker CD19. Thus, the main effect of this drug is the reduction of B cells, especially class-swtched memory B cells that might have a high capacity to activate other cells involved in the pathogenesis of SSc.References[1]M. Wang et al., “Identification and Validation of Predictive Biomarkers to CD19- and BCMA-Specific CAR T-Cell Responses in CAR T-Cell Precursors,” Blood, vol. 134, no. Supplement_1, pp. 622–622, Nov. 2019, doi: 10.1182/blood-2019-122513.[2]S. Ebata et al., “Safety and efficacy of rituximab in systemic sclerosis (DESIRES): a double-blind, investigator-initiated, randomised, placebo-controlled trial,” Lancet Rheumatol., 2021.AcknowledgementsThis work was funded by a grant from JDRF, LRA and NMSS (grant key: 2-SRA-2021-1043-S-B) and the Austrian Federal Government within the COMET K1 Centre Program, Land Steiermark and Land Wien.Disclosure of InterestsBarbara Dreo: None declared, Barbara Prietl: None declared, Selina Kofler: None declared, Verena Pfeifer: None declared, Harald Sourij: None declared, Florentine Moazedi-Fürst: None declared, Sonja Kielhauser: None declared, Monica D’Orazio: None declared, Sabine Zenz: None declared, Jens Thiel Speakers bureau: Novartis, GSK, Vifor, BMS, Consultant of: Novartis, GSK, Vifor, Grant/research support from: BMS, Martin Stradner: None declared, Hans-Peter Brezinsek: None declared

BackgroundAnti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitides (AAV) include granulomatosis with polyangiitis (GPA) and microscopic polyangiitis (MPA) (1, 2). ANCA recognize the antimicrobial proteins proteinase 3 (PR3) or myeloperoxidase (MPO) (1,2) and trigger the formation of neutrophil extracellular traps (NETs), which release DNA into the extracellular space and systemic circulation. This cell-free (cf)DNA induces endothelial damage, vascular inflammation and necrosis (3).ObjectivesThe nature, diagnostic and prognostic value of cfDNA in AAV is still unknown. The aim of the present study was to examine the clinical utility of cfDNA quantification as a biomarker in AAV.MethodsTotal DNA was isolated from platelet-free plasma samples of healthy controls (HC) and consecutive AAV patients. Plasma and clinical data were collected at baseline and follow-up. Copy numbers were quantified by qPCR for mtDNA (ATP-6 gene) and nuclear (n) DNA (GAPDH gene) (4). Patients with eosinophilic GPA (EGPA) were excluded.ResultsNinety-two HC (median age 51 ± 9, 48.2% female) and 104 AAV patients (median age 64 ± 10, 48% female, mean BVAS: 0; range: 0-40) were available for analysis. Eighty-four (80.8%) of these patients were diagnosed with GPA, and 20 with MPA (19.2%).mtDNA levels were significantly elevated in AAV plasma (8.7x107 copies/ml plasma, 95% CI: 5.3x107 to 1.3 x108)), compared to HC plasma (6.7x106 copies/ml plasma, 95% CI: 5.4x106 to 9.1x107, p6 copies/ml plasma, 95%CI: 2.7x106 to 5.0x106) and HC (3.3x106 copies/ml plasma, 95%CI: 2.4x106 to 4.7x106, p=0.30). ROC analysis showed that a cut-off value of 1.3x107 mtDNA copy numbers differentiated between AAV and HC with 89.4% sensitivity, 82.6% specificity and an AUC of 0.94. For AAV patients with active AAV, a cut-off value of 2.9x107 mtDNA copy numbers differentiated between AAV and HC with 96.1% sensitivity, 98.9% specificity and an AUC of 0.99 (Figure 1a).Figure 1.(a.) ROC curve for mtDNA plasma concentrations to discriminate between HC and active AAV patients (BVAS>0). AUC: area under the curve. (b.) Plasma mtDNA levels distinguish between AAV patients with active disease versus patients in a state of remission. Whiskers represent 95% CI. (c.) Plasma mtDNA levels in AAV patients correlate with the evolution of disease activity at follow-up.With the exception of the peripheral nervous system involvement, there was no association of mtDNA elevation with any particular type of active organ involvement at the time of blood sampling. A positive correlation between all cell-free DNA species and anti-MPO antibody titres was observed, as expected (for cfDNA, nDNA and mtDNA - r=0.25, p=0.01; r=0.21, p=0.02; r=0.22, p=0.02, respectively).AAV patients with active disease (BVAS>0) had a mean of 2.0x108 copies/ml of mtDNA in plasma which was higher compared to HC (p7copies/ml, p=0.03). For nDNA on the other hand, there were similar levels in active disease as in remission (5.3x106 and 4.8x106 copies/ml, respectively; p=0.64) (Figure 1b).Follow-up data were available for 27 AAV patients (median follow-up: 6 ± 6 months, IQR: 12). Longitudinal changes in mtDNA levels robustly correlated with changes in BVAS (r=0.56, p=0.002, Figure 1c).ConclusionThe quantification of cell free mtDNA - but not nDNA - copy numbers allows a sensitive and specific distinction between healthy individuals and patients with active AAV. mtDNA levels correlate cross sectionally with disease activity in AAV patients. Plasma mtDNA quantification may therefore aid in the diagnosis of AAV and in monitoring AAV activity.References[1]Kitching, A.R., et al. Nat Rev Dis Primers6, 71 (2020).[2]Kallenberg, C. Nat Rev Rheumatol10, 484–493 (2014).[3]Kessenbrock, K., et al. Nat Med15, 623–625 (2009).[4]Giaglis S, et al. RMD Open 2021;7:e002010.(2021)Disclosure of InterestsNone declared

BackgroundRheumatoid arthritis (RA) is a systemic autoimmune disease leading to erosive joint destruction. Although the exact pathogenesis is still elusive, the strong association of certain HLA class II molecules, such as HLA-DRB1*0401 (HLA-DR4), suggest involvement of CD4+ T cells (1,2). Mouse models of RA mimic specific aspects of the disease but are limited by the differences between human and murine immune systems.ObjectivesWe aimed to establish a humanized mouse model (humice) carrying DR4+ RA PBMCs to study its role in the pathogenesis of RA without putting patients at risk.MethodsPeripheral blood mononuclear cells (PBMC) of HLA-DR4 positive RA patients or controls were isolated and injected into NSG-Ab0 DR4 mice (NOD-scid IL2Rgammanull mice lacking MHC class II while expressing the human HLA-DR4) to create humice. Human immune cell composition within humice was profiled using flow cytometry. Development of RA was monitored by examination of the joints and micro computed tomography analysis. Joints were analysed by histology regarding pannus formation, bone erosions, cartilage damage, and human cell infiltration.ResultsTransfer of RA PBMCs induced arthritis in humice recapitulating hallmarks of RA including immune cell infiltration, pannus formation, increased osteoclastogenesis, cartilage damage, and bone erosions. Arthritis was dependent on the implanted human cells as NSG-Ab0 DR4 mice without transfer of human PBMCs did not develop arthritis. T-helper 1 (Th1) cells, dominated the human immune cell composition in humice, while regulatory T cells (Tregs) were diminished compared to donor PBMC composition. Mice humanized with cells from RA patients were more likely to develop inflammatory joint disease, compared to healthy HLA-DR4 positive controls (RA donor 70% vs. healthy control 20%, p=0.00196). CTLA-4 Ig treatment prevented arthritis development in this model (p=0.0055).ConclusionHumice carrying DR4+ RA PBMCs developed an RA-like erosive joint disease driven by the implanted human immune system. The data implies that the disease can be transferred by arthritogenic cells found in the peripheral blood of RA patients. This model will allow new insights into the pathogenesis of RA.References[1]Goulielmos GN, Zervou MI, Myrthianou E, Burska A, Niewold TB, Ponchel F. Genetic data: The new challenge of personalized medicine, insights for rheumatoid arthritis patients. Gene Available from: http://www.ncbi.nlm.nih.gov/pubmed/26869316[2]Holoshitz J. The rheumatoid arthritis HLA–DRB1 shared epitope. Curr Opin Rheumatol Available from: http://www.ncbi.nlm.nih.gov/pubmed/20061955Disclosure of InterestsNone declared.

BackgroundLittle is known about glucocorticoid-sparing agents in giant cell arteritis (GCA) except for IL-6 inhibition. Secukinumab (SEC) has shown significant improvements in the signs and symptoms of IL-17A driven medical conditions such as plaque psoriasis, psoriatic arthritis, and axial spondyloarthritis.1,2 It has a favourable long-term safety profile.1,2ObjectivesTitAIN is the first randomised controlled trial investigating the potential efficacy, safety, and tolerability of SEC in GCA patients (pts).MethodsThis phase 2, randomised, double-blind, placebo (PBO) controlled, multicentre, proof-of-concept trial enrolled pts (aged ≥50 years) with new onset (diagnosed within 6 weeks (wks) of baseline) or relapsing (diagnosed >6 wks from baseline) GCA, naïve to biological therapy. Pts were randomised (1:1) to SEC 300 mg or PBO initially administered wkly (5 doses) and every 4 wks thereafter through Wk 48 (last dose), in combination with a 26-wk prednisolone taper regimen starting from baseline. Proportion of GCA pts in sustained remission until Wk 28 was the primary endpoint assessed by a Bayesian analysis of the posterior distribution with non-responder imputation. Other key endpoints included proportion of GCA pts in sustained remission until Wk 52 (based on study data with non-responder imputation) and time to first GCA flare after baseline.ResultsOut of 52 randomised pts (SEC, n=27; PBO, n=25), 71.2% (n=37) completed study treatment (SEC, 81.5%; PBO, 60.0%). Overall, 42 (80.8%) pts had new onset GCA and 10 (19.2%) pts had relapsing GCA at baseline. Proportion (posterior median with 95% credibility interval) of GCA pts in sustained remission until Wk 28 was higher with SEC, 70.1% (51.6%-84.9%), than with PBO, 20.3% (12.4%-30.0%); odds ratio (posterior median with 95% credibility interval), 9.31 (3.54-26.29) (Table 1). Until Wk 52, proportion (95% confidence interval) of GCA pts in sustained remission were 59.3% (38.8%-77.6%) in SEC group and 8.0% (1.0%-26.0%) in PBO group (Table 1). Median (95% confidence interval) time to first GCA flare after baseline was not reached for GCA pts treated with SEC and was 197.0 (101.0-280.0) days for PBO (Figure 1). Overall, treatment-emergent adverse events (AEs) occurred in 98.1% (SEC vs PBO, 100.0% vs 96.0%) and serious AEs in 32.7% (SEC vs PBO, 22.2% vs 44.0%) pts. Two pts in each SEC and PBO groups had AEs that led to study drug discontinuation and 1 pt in each group had AEs that led to death (not treatment-related). There were no new or unexpected safety signals identified with SEC treatment.Table 1.Proportion of GCA patients with sustained remission (Full analysis set) until Week 28 and 52Proportion of ptsSecukinumab (N=27)Placebo (N=25)Median percentage (95% credibility interval), Wk 2870.1% (51.6%, 84.9%)20.3% (12.4%, 30.0%)Percentage (95% confidence interval), Wk 5259.3% (38.8%, 77.6%)8.0% (1.0%, 26.0%)The full analysis set comprises all pts to whom study treatment has been assigned by randomisation and who received at least one dose of randomised study treatment (secukinumab or placebo).GCA, giant cell arteritis; N, number of pts in each treatment group in the full analysis set, pts, patients; Wk, WeekFigure 1.Kaplan-Meier plot of time to first GCA flare from baseline up to Week 52 (Full analysis set)ConclusionSEC demonstrated a higher sustained remission rate and longer time to first GCA flare vs PBO through 52 wks in pts with GCA. This proof-of-concept phase 2 study supports further development of SEC as a potential treatment in combination with 26 wk glucocorticoid taper for pts with GCA.References[1]Mease PJ, et al. ACR Open Rheumatol. 2020;2(1):18-25[2]Baraliakos X, et al. RMD Open. 2019;5:e001005Disclosure of InterestsNils Venhoff Speakers bureau: AbbVie, Novartis, Bristol-Myers-Squibb, Chugai, Roche, Vifor, Consultant of: AbbVie, Chugai, Novartis, Vifor, Grant/research support from: Bristol-Myers-Squibb, Novartis, Wolfgang A. Schmidt Speakers bureau: Abbvie, Chugai, Medac, Novartis, Roche, Sanofi, Consultant of: Advisory board member: Abbvie, Chugai, GlaxoSmithKline, Novartis, Roche, Sanofi, Grant/research support from: principle investigator in GCA trials: Abbvie, GlaxoSmithKline, Novartis, Sanofi, Raoul Bergner Speakers bureau: Abbvie, Bristol Myers Squibb, Chugai, Novartis, Roche, Consultant of: Advisory board member: Gilead, GlaxoSmithKline, Vifor, Jürgen Rech Speakers bureau: Abbvie, Biogen, BMS, Chugai, GSK, Janssen, Lilly, MSD; Novartis, Roche, Sanofi, Sobi, UCB, Consultant of: Abbvie, Biogen, BMS, Chugai, GSK, Janssen, Lilly, MSD, Novartis, Roche, Sanofi, Sobi, UCB, Leonore Unger Paid instructor for: Novartis, Hans-Peter Tony Consultant of: Abbvie, BMS, Chugai, Gilead, Lilly, Novartis, Roche, Sanofi, Meryl Mendelson Shareholder of: Novartis Pharmaceuticals Corporation, Employee of: Novartis Pharmaceuticals Corporation, Christian Sieder Employee of: Novartis Pharma GmbH, Meron Maricos Employee of: Novartis Pharma GmbH, Jens Thiel Speakers bureau: Novartis, GSK, Bristol-Myers-Squibb, Roche, AstraZeneca, Vifor, Consultant of: Novartis, Janssen, GSK; research grants: Bristol-Myers-Squibb, Novartis

BackgroundPrimary Sjögren’s Syndrome (pSS) is a chronic autoimmune disease. Symptoms range from sicca to systemic, potentially life-threatening organ damage. Little is known about the onset of the disease. Anti-Ro antibodies are described to develop years before the first symptoms. In addition, first degree relatives of pSS patients have an 11- to 19- fold increased risk of developing pSS themselves.ObjectivesTo identify and follow-up individuals at risk for pSS in order to study symptoms and immune pathology before and at development of pSS.MethodsIn this ongoing long-term study individuals at risk for developing pSS but not fulfilling the ACR-EULAR classification criteria of pSS were included, defined as: 1.) Anti-SSA positive individuals (Anti-SSA+) without any sicca symptoms or diagnosis of an underlying systemic autoimmune disease; 2.) First degree relatives of patients (relatives) with an established diagnosis of pSS and typical autoantibodies (ANA ≥ 1:160 and/or anti-SSA+ and/or rheumatoid factor+); 3.) Individuals with at least one feature of the ACR-EULAR classification criteria for pSS, but not fulfilling the criteria (incomplete).At baseline and at annual visits, demographic data, blood, saliva and urine samples were collected and stored. Salivary and lacrimal flow, salivary gland ultrasonography (SGUS), and patient-related outcome measures were analysed. A lip salivary gland biopsy was performed at baseline and upon development of symptoms suggestive of pSS. The primary endpoint was the development of definite pSS according to the ACR-EULAR classification criteria.ResultsAfter the first year of recruitment, 50 individuals (Anti-SSA+ n=27, relatives n=21, incomplete n=2) were screened at baseline, of whom 28 were identified as individuals at risk for pSS and were included in the study. Twenty-two individuals were excluded from the study, most of whom were “relatives” with negative autoantibodies. Of these 28 individuals at risk, 89% were female (n=25), they had a median age of 53 years (IQR: 19) and 57% (n=16) had positive antinuclear antibodies. 86 percent were positive for anti-SSA and 14% were positive for anti-SSB. Decreased complement C3 and C4 were found in 18% and 4%, respectively. Serum IgG concentration was elevated in 29% of individuals. A reduction of lacrimal flow was found in 29% and stimulated whole salivary flow was reduced in 29%. The median of the ESSPRI was 1.6 (3.0). Eight-teen percent of the investigated individuals had a pathological ultrasound [Hocevar score median 4,5 (9,0)] and in 9% a focus score ≥ 1 [median 0.15 (0.57)] was found in the lip salivary gland biopsies. Four patients (14%) met the primary endpoint and were diagnosed with pSS within the first year.ConclusionThe design of the PRESTIGE study allows us to follow individuals at risk for pSS and will help to unveil symptoms and immune pathology as pSS develops. We suggest to establish a larger international pre-pSS cohort to increase statistical power.Disclosure of InterestsNone declared

BackgroundThe most frequent form of vasculitis in elderly people is giant cell arteritis (GCA) with an annual incidence rate less than 10 per 100,000 persons over the age of 50. Like most vasculitides, GCA is characterized by chronicity and relapses, leading to significant overall morbidity and higher mortality in a subset of patients with aortic involvement and dissection. Most studies carried out so far have been retrospective, used monocentric study designs and small patient cohorts. Therefore, the Joint Vasculitis Registry in German-speaking countries (GeVas) has been established to record patients, who have been recently diagnosed with vasculitis or who have changed their treatment due to a relapse (inception cohort). The GeVas-Registry allows a long-term follow-up of a substantial cohort of vasculitis patients in a prospective and multicenter manner.ObjectivesTo describe the subgroup of GCA and its characteristics within the GeVas registry.MethodsGeVas is a prospective, web-based, multicenter, clinician-driven registry for the documentation of organ manifestations, damage, long-term outcomes, and therapy regimens in various types of vasculitis. Recruitment started in June 2019. By January 2022, 17 centers in Germany were initiated and have begun enrolling patients. Meanwhile, more than 350 patients have been documented in the registry. Sites in Austria and the German-speaking cantons of Switzerland will be integrated soon (1).ResultsBy mid-October 2021, the participating centers recruited 131 GCA patients into the registry. 21.7% of patients (n=28) were enrolled in the registry due to relapse, and 78.3% (n=101) due to a first-time diagnosis. In accordance with long-standing epidemiology data, the majority of patients (67,2%), were female (n=88), and 32.8% (n=43) were male. Mean age was 74 years (max. 92y, min. 52y). The most frequently recorded organ manifestations in GCA patients addressed cranial and ophthalmic symptoms, and the cardiovascular system. However, vascular lung/chest involvement was also observed in 3% of cases (n=4). Out of the 131 patients, 97.7% (n=128) received immunosuppressive therapy, three refused to take any medication. An equal number of patients were treated with glucocorticoid (GC) therapy. While about two equal parts were treated by stable long term oral GC therapy (47,7%, n=62) or by i.v. pulse therapy followed by tapering (49,2%, n=64), only about 2.3% (n=3) were treated by oral GC therapy with intermittent i.v. pulses. 48.5% (n=63) of patients received tocilizumab as additional immunosuppressive therapy, 19.2% (n=25) methotrexate, and 18.5% (n=24) cyclophosphamide i.v. pulses.ConclusionIn June 2019, we successfully established the prospective multicenter vasculitis registry GeVAS. It describes the first systematically recorded prospective GCA cohort in German-speaking countries. Its characteristics correspond to those that can be expected from the literature, with some unexpected finding e.g. the high proportion of patients treated with cyclosphosphamid, probably reflecting a sicker patient population with e.g. aortic or central nervous involvement. After 2.5 years of follow-up documentation, the first long-term results will be systematically evaluated and interpreted. The newly acquired data on disease manifestation, diagnostics and therapy regimens will provide important insights into the treatment of GCA patients in Germany and may generate further research goals.ReferencesTrial registration: German Clinical Trials Register (Deutsches Register Klinischer Studien): DRKS00011866. Registered 10 May 2019. 3[1]C Iking-Konert; P Wallmeier; S Arnold; S Adler; K de Groot; B Hellmich; B Hoyer; K Holl-Ulrich; Ihorst; M Kaufmann; I Kötter; U Müller-Ladner; T Magnus; J. Rech; H. Schulze-Koops; N. Venhoff; T. Wiech; P. Villiger; F. Schubach; P. Lamprecht. The Joint Vasculitis Registry in German-speaking countries (GeVas) – a prospective, multicenter registry for the follow-up of long-term outcomes in vasculitis. BMC Rheumatol. 2021 Jul 31;5(1):40. doi: 10.1186/s41927-021-00206-2.AcknowledgementsGeVas was supported by unrestricted grants by: DGRh, John Grube Foundation, Vifor and Roche PharmaDisclosure of InterestsPia Wallmeier: None declared, Sabrina Arnold: None declared, Fabian Schubach: None declared, Gabriele Ihorst: None declared, Peer Aries: None declared, Raoul Bergner Consultant of: Advisory Board VIFOR, Grant/research support from: John-Grube Research Award 2021, Jan Philip Bremer: None declared, Norman Görl: None declared, Bernhard Hellmich: None declared, Jörg Henes: None declared, Bimba Hoyer: None declared, Antje Kangowski: None declared, Ina Kötter: None declared, Tim Magnus: None declared, Claudia Metzler: None declared, Ulf Müller-Ladner: None declared, Matthias Schaier: None declared, Ulf Schönermark: None declared, Jens Thiel: None declared, Leonore Unger: None declared, Nils Venhoff Speakers bureau: Roche and Vifor, Consultant of: Roche and Vifor, Grant/research support from: John-Grube Research Award 2021, Julia Weinmann-Menke: None declared, Jana Petersen: None declared, Peter Lamprecht Speakers bureau: Lecture fees from: Chugai, GSK, Roche, Consultant of: Consulting & lecture fees from: Chugai, GSK, Roche, and Vifor., Grant/research support from: Research grants for GeVas: DGRh, John Grube Foundation, Roche, and Vifor, Christof Iking-Konert Speakers bureau: lecture fees from: Chugai, GSK, Roche, and Vifor., Consultant of: Consulting fees from: Chugai, GSK, Roche, and Vifor., Grant/research support from: Research grants for GeVas: DGRh, John Grube Foundation, Roche, and Vifor

BackgroundImmunocompromised patients are considered high-risk and prioritized for vaccination against COVID-19 (1). Furthermore, vaccination-induced CD4 and CD8 T-cell responses have been suggested to have a protective role in COVID-19 (2). If T-cell responses are diminished after vaccination in immunocompromised individuals is not known to date.ObjectivesTo investigate cellular immunity following mRNA vaccination against COVID-19 in healthy individuals and patients undergoing B-cell depletion therapy.MethodsIn this interim analysis of the CoVVac study (NCT04858607), we analyzed T-cell responses in autoimmune patients treated with B-cell depleting therapy (BD, n=41) and age-matched healthy controls (HCs, n=50) 3-4 weeks after the second dose of mRNA vaccination against COVID-19. Therefore, we isolated PBMCs and stimulated them with a peptide pool covering the spike protein in vitro. Reactive CD4 and CD8 T-cells were determined by staining for IFNg, TNFa, IL-2 and GzmB by flow cytometry. Anti-SARS-CoV-2 antibody assays targeting the receptor-binding domain (RBD) or trimeric S protein (TSP) were performed to elucidate concomitant B-cell responses.ResultsWe observed significant alterations in anti-SARS-CoV-2 antibody responses in our cohort, the frequency of IFNg+ and IL-2+ CD4 and CD8 T-cells was similar in BD patients and controls. On the other hand, TNFa+ CD4 T-cells were significantly enriched in healthy controls versus BD patients (p=0.017) and correlated significantly with antibody titres (p=0.003). Similarly, GzmB+ CD8 T-cells were significantly diminished in our patient cohort (pOnly 21 (42%) healthy individuals and 14 (34%) patients showed reactive T-cells for all the cytokines tested. This observation is mainly explained by a lack of cytokine production of CD8 T-cells in 26 (52%) HCs and 27 (66%) BD patients. In turn, 22 (44%) HCs and 17 (42%) patients didn’t show any IL-2 producing CD8 cells. Of note, only 2 (4%) of HCs showed no GzmB+ CD8 T-cells whereas the number increased to 15 (37%) of BD individuals (pConclusionOur data suggest that most patients with B-cell depleting therapy are able to mount T-cell responses similar to those of healthy individuals while a minority of these patients did not show complete immunity against SARS CoV-2. Further analyses are needed to better understand a possible link of B-cell depletion therapy and CD8 T-cell responses.References[1]Shields AM, Burns SO, Savic S, Richter AG; UK PIN COVID-19 Consortium. COVID-19 in patients with primary and secondary immunodeficiency: The United Kingdom experience. J Allergy Clin Immunol. 2021 Mar;147(3):870-875.e1.[2]Angyal, A., Longet, S., Chalk, J., 2022. T-cell and antibody responses to first BNT162b2 vaccine dose in previously infected and SARS-CoV-2-naive UK health-care workers: a multicentre prospective cohort study. The Lancet Microbe 3Disclosure of InterestsNone declared

BackgroundThe Interstitial lung disease (ILD) represent a major cause of mortality in Systemic sclerosis (SSc). Validated methods for screening and therapy monitoring of SSc-ILD are needed. In the recent literatur, the transthoracic lung sonography (LUS) emerges as a reliable tool for the early detection of lung alterations. It was recently reported that SSc-ILD-patients have thicker artefacts reflecting the pleuraline compared with SSc-patients without ILD, but it still remains unknown if the thickness of the pleuraline changes with a proper therapy response.ObjectivesThe aim of the study was to compare the thickness of the pleuraline of SSc-ILD-patients without biological Disease Modifying Anti-Rheumatic Drugs (bDMARDS) with those successfully treated with bDMARDs in order to establish if LUS can be a reliable surveillance tool in SSc-ILD. Additionally, since our Grazer-Schema (Mycofenolat mofetil (MMF)+ rituximab (RTX) 500 mg i.v. in week 0 and 2 every 3 months) is succesfully used in our clinic since more than 10 years in patients nonresponding to methotrexate (MTX) or cyclophosphamide, we report 10 years follow-up data of 5 Patients before and after receiving that schema.Methods29 prospectively enrolled SSc-patients were assessed according to the European League Against Rheumatism Scleroderma Trial and Research standards. Written informed consent was obtained from all patients and the study was approved by the Ethics Committee of Graz. Validated clinical scores, such as the scleroderma disease activity (SScAS) and the scleroderma disease severity scale (SScSS) were assessed. The thickness of the pleuraline was detected with LUS. The presence of ILD was assessed by high-resolution computed tomography. Twenty-five SSc-patients enrolled between 2008 and 2009, who underwent LUS with the same technical instruments and some of the same operators, were taken as a control group since they were untreated yet or were not assuming bDMARDs(1).ResultsOf the 29 SSc-patients (27 females/2 males) enrolled between 2019 and 2020, seventeen patients had radiographic signs of ILD (SSc-ILD+/2019-20, n=17). Until the LUS was performed, all SSc-ILD+/2019-20 showed a good clinical response to the therapy with RTX in combination with MMF. Of the 25 SSc-patients enrolled in our previous study, (n=25; 23 females/ 2 males), twelve of these patients had radiographic signs of ILD (SScILD+/2009-10, n=12); 7 were under MTX or Cyclosporin and 5 were untreated. Additionally, data collected from 5 SSc-ILD+/2009-2010 at baseline were analysed in a 10- years- follow up. As expected, SSc-ILD+ patients had a thicker pleuraline than SSc-ILD- patients. The pleura was significantly thinner in patients under MMF und RTX compared to the pleuraline of patients without bDMARDs (1,8 mm ± 0,7 Vs 0,95 ± 0,31; p < 0,00). Interestingly those patients reported less frequently dry cough or dyspnoea, showing that the lower pleura thickness could be associated with a better clinical outcome. In a 10 Year follow-up after continued treatment with MMF and RTX, a significant reduction of the thickness of the pleura could be found (Mean 0,68 ± 0,2 VS 1,5 ± 0,4; p < 0,00). Interestingly, such significant improvement could be found in all of these 5 patients receiving our Grazer Schema. Remarkably, the pleuraline-thickness was comparable to the thickness of the pleura of SSc-ILD-patients. This finding shows that the increase in thickness of the pleuraline can be totally reversible under treatment.ConclusionDue to the encouraging validity, reliability and simplicity of LUS, it has been increasingly considered as an excellent screening tool for SSc-ILD. Its sensitivity to reveal parenchyma and pleura changes over time in treated patients, make it also a helpful and safe methodology to follow up SSc-ILD patients.References[1]Moazedi-Fuerst, F. C. et al. Pulmonary echography in systemic sclerosis. Clinical Rheumatology (2012) doi:10.1007/s10067-012-2055-8.Disclosure of InterestsNone declared

BackgroundGranulomatosis with polyangiitis (GPA) is the second most frequent vasculitis in Germany with an annual incidence of 34 per million and a prevalence of 210 per million [1]. GPA is characterized by its chronic course, frequent relapses, significant overall morbidity and mortality, and substantial socio-economic impact. Multiorgan involvement affecting the respiratory tract, kidney, and other organs is common. Limited variants also occur [2]. So far, prospective long-term observational data on the disease course of GPA are missing in Germany. Therefore, the Joint Vasculitis Registry in German-speaking countries (GeVas) has been established to follow the course of patients recently diagnosed with vasculitis or a change of their treatment due to a relapse (inception cohort). The GeVas registry allows long-term follow-up of a substantial cohort of vasculitis patients in a multicenter setting.ObjectivesTo present the first data on the follow-up of newly diagnosed and relapsing GPA enrolled in the GeVas registry.MethodsGeVas is a prospective, web-based, multicenter, clinician-driven registry for the documentation of organ manifestations, damage, long-term outcomes, and therapy regimens in various types of vasculitis. Recruitment started in June 2019. By January 2022, 17 centers in Germany were initiated and started enrolling patients. Meanwhile, more than 350 patients have been documented in the registry. Sites in Austria and the German-speaking cantons of Switzerland will be integrated soon [3].ResultsBy mid-October 2021, the participating centers included 113 patients with GPA. The majority of patients were PR3-ANCA positive and affected by general symptoms, ENT, lung, renal, and neurological involvement. Patients commonly received cyclophosphamide or rituximab in combination with glucocorticoids for the induction of remission. Fewer patients received methotrexate or other immunosuppressants. Patient characteristics and therapy are summarized in Table 1.Table 1.Patient characteristics (n = 113). *Unless otherwise specified.CategoryFeaturen (%)*AgeAge (years); median [range]60 [51 - 70]GenderMale61 (54.0)Female52 (46.0)Reason for inclusion in the registryNewly diagnosed vasculitis57 (51.4)Relapse56 (49.6)ANCA statusPR3-ANCA99 (87.6)MPO-ANCA4 (3.6)ANCA negative9 (7.9)Organ manifestationGeneral symptoms86 (76.1)ENT69 (61.1)Lung/chest66 (58.4)Renal35 (31.0)Cardiovascular7 (6.2)GI3 (2.7)Neurological27 (23.9)TherapyGlucocorticoid102 (90.3)Rituximab56 (49.6)Cyclophosphamide37 (32.7)Methotrexate and other immunosuppressants, respectively26 (23.0) and 19 (16.8), respectivelyConclusionHere, we present the first interim analysis of the GeVas registry. Clinical manifestations of GPA reported herein show less frequent renal involvement in comparison with a recent report from another European registry (POLVAS) and an UK study [4, 5]. This is potentially related to the predominance of recruiting rheumatology centers thus far. By contrast, respiratory tract involvement is more frequent and PR3-ANCA less common in Japan [5]. Further data are prospectively documented and a follow up analysis is in progress.References[1]Hellmich B, et al. New insights into the epidemiology of ANCA-associated vasculitides in Germany: results from a claims data study. Rheumatology 2021;60:4868-73.[2]Kitching AR, et al. ANCA-associated vasculitis. Nat Rev Dis Primers 2020;6:71.[3]Iking-Konert C, et al. The Joint Vasculitis Registry in German-speaking countries (GeVas) – a prospective, multicenter registry for the follow-up of long-term outcomes in vasculitis. BMC Rheumatol 2021;5:40.[4]Wójcik K, et al. Clinical characteristics of Polish patients with ANCA-asscoiated vasculitides – retrospective analysis of POLVAS registry. Clin Rheumatol 2019;38:2553-63.[5]Furuta S, et al. Comparison of the phenotype and outcome of granulomatosis with polyangiitis between UK and Japanese cohorts. J Rheumatol 2017;44:216-22.AcknowledgementsGeVas was supported by unrestricted grants by: DGRh, John Grube Foundation, Vifor and Roche PharmaDisclosure of InterestsSabrina Arnold: None declared, Pia Wallmeier: None declared, Fabian Schubach: None declared, Gabriele Ihorst: None declared, Peer Aries: None declared, Raoul Bergner Consultant of: VIFOR, Jan Philip Bremer: None declared, Norman Görl: None declared, Bernhard Hellmich: None declared, Jörg Henes: None declared, Bimba Hoyer: None declared, Antje Kangowski: None declared, Ina Kötter: None declared, Claudia Metzler: None declared, Ulf Müller-Ladner: None declared, Matthias Schaier: None declared, Ulf Schönermark: None declared, Jens Thiel: None declared, Leonore Unger: None declared, Nils Venhoff Speakers bureau: Roche and Vifor: speaker honoraries, Consultant of: Roche and Vifor: advisory boards, Grant/research support from: John-Grube Research Award 2021, Julia Weinmann-Menke: None declared, Jana Petersen: None declared, Christof Iking-Konert Speakers bureau: Lecture fees from: Chugai, GSK, Roche, and Vifor, Consultant of: Consulting fees from: Chugai, GSK, Roche, and Vifor, Grant/research support from: Research grants for GeVas: Roche, Vifor, DGRh, John Grube Foundation, Peter Lamprecht Speakers bureau: Chugai, GSK, Roche, and Vifor, Consultant of: Chugai, GSK, Roche, and Vifor, Grant/research support from: DGRh, John Grube Foundation, Roche, and Vifor

BackgroundNumerous cytokines that influence disease activity in psoriatic arthritis (PsA) are modulators of the Janus Kinases/Signal Transducers and Activators of Transcription (JAK/STAT) pathway. The JAK1/STAT1/STAT3/STAT5 network can drive the expansion of Th17 and regulatory T cells via proinflammatory cytokines in PsA joints,[1], [2] while hyperphosphorylation of STAT3 in immune cells has previously been shown to promote PsA pathogenesis through the Interleukin (IL)-23/IL-17/IL-22 axis.[3] Therefore, the phosphorylation status of STAT molecules in leucocytes of PsA patients may indicate active disease and could potentially guide treatment with JAK inhibitors.ObjectivesTo analyse phosphorylated STAT (pSTAT) levels of circulating leucocyte subsets in PsA patients with active and inactive diseaseMethodsWhole blood was drawn on consecutive PsA patients fulfilling the CASPAR criteria[4] to perform flow cytometry analysis using the BD FACSLyric platform. Disease activity was assessed using the Disease activity for psoriasis arthritis (DAPSA) score.[5] All steps from storage of drawn blood to cell fixation were performed at 4°C to prevent auto-activation of leucocytes. The geometric mean fluorescence intensities (gMFI) of pSTATs in granulocytes, monocytes, B cells and CD4+/- naïve/memory T cells were compared between patients with moderate to high (MoDA/HDA) and remission to low disease activity (REM/LDA). Correlation analysis between gMFIs and DAPSA scores were performed.ResultsForty-two patients (female ratio: 0.48) with established PsA (median ± standard deviation, age: 56 ± 12.54 years, disease duration: 8.50 ± 7.10 years) were included in this study. Twenty-one percent of patients were in MoDA/HDA, while the remaining 79% were in REM/LDA. Patients in MoDA/HDA showed significantly higher pSTAT3 levels in CD4+ naïve (gMFI median ± standard deviation: 284.5 ± 79.9 vs 238 ± 92.9, p = 0.011), CD4- naïve (297 ± 107.5 vs 238 ± 98.4, p = 0.04), CD4+ memory (227 ± 62.9 vs 190.5 ± 72.2, p = 0.009) and CD4- memory T cells (209 ± 66.8 vs 167.0 ± 64.9, p = 0.036). On the other hand, PsA patients in remission or low disease activity displayed higher pSTAT1 levels in granulocytes (2509 ± 1887 vs 1330.5 ± 784.1, p = 0.040) and monocytes (255 ± 230 vs 144 ± 62.5, p = 0.049). Positive correlations were found between DAPSA scores and pSTAT3 in CD4+ naïve and memory T cells (Spearman’s correlation coefficient rho (ρ) = 0.5, p = 0.0012 and ρ = 0.47, p = 0.0025 resp.) whereas pSTAT1 in granulocytes and monocytes were negatively correlated with the DAPSA scores (ρ = -0.45, p = 0.0074 and ρ = -0.34, p = 0.05).ConclusionDifferential phosphorylation of STAT3 and STAT1 molecules in circulating leucocyte subsets indicates PsA disease activity. Further studies to examine the value of STAT phosphorylation patterns guiding JAK inhibitor therapy are underway.References[1]U. Fiocco et al., “Ex vivo signaling protein mapping in T lymphocytes in the psoriatic arthritis joints,” J. Rheumatol., vol. 93, pp. 48–52, 2015, doi: 10.3899/jrheum.150636.[2]S. K. Raychaudhuri, C. Abria, and S. P. Raychaudhuri, “Regulatory role of the JAK STAT kinase signalling system on the IL-23/IL-17 cytokine axis in psoriatic arthritis,” Ann. Rheum. Dis., vol. 76, no. 10, pp. e36–e36, 2017.[3]E. Calautti, L. Avalle, and V. Poli, “Psoriasis: A STAT3-centric view,” International Journal of Molecular Sciences, vol. 19, no. 1. MDPI AG, Jan. 06, 2018, doi: 10.3390/ijms19010171.[4]W. Taylor, D. Gladman, P. Helliwell, A. Marchesoni, P. Mease, and H. Mielants, “Classification criteria for psoriatic arthritis: Development of new criteria from a large international study,” Arthritis Rheum., vol. 54, no. 8, pp. 2665–2673, 2006, doi: 10.1002/art.21972.[5]M. M. Schoels, D. Aletaha, F. Alasti, and J. S. Smolen, “Disease activity in psoriatic arthritis (PsA): Defining remission and treatment success using the DAPSA score,” Ann. Rheum. Dis., vol. 75, no. 5, pp. 811–818, 2016, doi: 10.1136/annrheumdis-2015-207507.Disclosure of InterestsBarbara Dreo: None declared, Daniel Ruben Pietsch: None declared, Rusmir Husic Speakers bureau: MSD, Lilly und Abbvie, Angelika Lackner: None declared, Johannes Fessler: None declared, Janine Rupp: None declared, Anirudh Subramanian Muralikrishnan: None declared, Jens Thiel Speakers bureau: GSK, BMS, AbbVie, Novartis, Consultant of: GSK, Novartis, Grant/research support from: BMS, Martin Stradner Speakers bureau: Eli Lilly, Pfizer, MSD, BMS, AbbVie, Janssen, Consultant of: Eli Lilly, AbbVie, Janssen, Philipp Bosch Grant/research support from: Pfizer

BackgroundAxial spondyloarthritis (axSpA) and psoriatic arthritis (PsA) may have a profound impact on health-related quality of life (HRQoL) and sleep despite effective treatment.ObjectivesTo assess sleep and HRQoL in SpA and determine associated factors.MethodsMonocentric questionnaire-based assessment of HRQoL, function, sleep and depression in 314 SpA patients (n=168 PsA, n=146 axSpA).ResultsUnder effective treatment 138 SpA patients (46.5%) demonstrated abnormal sleep behaviour. 49.3% reported not being able to sleep through the night, with 6.1 % needing sleeping pills. 11.9% indicated feeling unrefreshed most mornings. Abnormal sleep behaviour was associated with female sex (p=0.005), HLAB27 (p=0.034), functional impairment (p=0.001) and depression (pConclusionDespite treatment many SpA patients have a reduced HRQoL and impaired sleep quality with significant differences between male and female patients. Impact of COVID19 pandemic was low.Table 1.Patient characteristicsTotal (n=314)Women n=142 (45.2%)Men n=172 (54.8%)p-valueAge, years, mean (SD)53.8 (13.9)53.7 (14.5)53.8 (13.4)0.983BMI, kg/m2, mean (SD)27.3 (6.3)27.1 (6.5)27.4 (6.1)0.706Smokers, n (%)68 (23.1)34 (24.6)34 (21.7)0.544HLA B27, n (%) (n=230)120 (52.2)46 (45.1)54 (54.9)0.055CRP, mg/l, mean (SD)6.14 (11.9)5.88 (6.0)6.37 (15.2)0.721Significant functional impairment, n (%)54 (17.5)29 (20.6)25 (15.0)0.198Therapy, n (%) csDMARD113 (36.2)56 (39.7)57 (33.3)0.243 bDMARD211 (67.4)91 (64.1)120 (70.2)0.252Depressive symptoms, n (%) Mild109 (37.5)53 (39.3)56 (35.9)0.040 Moderate/severe48 (16.5)27 (20.0)21 (13.5)WHO-QOL mean (SD) Physical60.2 (19.4)57.4 (19.7)62.6 (19.0)0.019 Mental67.7 (17.2)64.5 (17.8)70.3 (16.2)0.003 Social67.8 (20.0)67.4 (19.6)68.2 (20.3)0.703 Environmental77.3 (13.3)76.6 (13.6)78.0 (13.1)0.363Sleep, n (%) Abnormal sleep behaviour138 (46.5)76 (55.1)62 (39.0)0.006 Inability to sleep through the night145 (49.4)76 (55.1)69 (44.2)0.024 Waking too early89 (30.4)43 (31.4)46 (29.5)0.459 Need for sleeping pills18 (6.1)11 (8.0)7 (4.4)0.133 Unrefreshing sleep most or all nights35 (11.9)24 (17.5)11 (7.0)Figure 1.Female SpA patients have increased depressive symptoms and a reduced HRQoL.Disclosure of InterestsNatalie Frede Grant/research support from: Novartis study grant, Eva Rieger: None declared, Raquel Lorenzetti Grant/research support from: Novartis study grant, Ana Venhoff: None declared, Carolin Hentze: None declared, Ilona Jandova: None declared, Cornelia Glaser: None declared, Jens Thiel Speakers bureau: Novartis, AbbVie, Pfizer, BMS, UCB, Consultant of: Novartis, AbbVie, Pfizer, BMS, UCB, Grant/research support from: BMS, Novartis study grants, Reinhard Voll Speakers bureau: Novartis, AbbVie, Pfizer, BMS, UCB, Consultant of: Novartis, AbbVie, Pfizer, BMS, UCB, Lilly, Grant/research support from: Novartis study grant, Nils Venhoff Speakers bureau: Novartis, AbbVie, Pfizer, BMS, UCB, Consultant of: Novartis, AbbVie, Grant/research support from: Novartis study grant

BackgroundPrevious studies have provided evidence that T cells may play a significant role in the pathogenesis of systemic sclerosis (SSc) and observational studies reported that Abatacept, which interferes with T cells activation, appeared to be safe and effective in SSc-patients with skin and muscle involvement. So far, there was no significant effective treatment for SSC-ILD. We have recently demonstrated the effectiveness of intensified –RTX- long term treatment (“Grazerprotocol” with 500 mg RTX in week 0 and 2, every 3 months +/- MMF) in SSc patients with severe organ involvement and/or progressive form non responding to methotrexate or cyclophosphamide. However, no data are available concerning how long Patients with a stable disease need to be treated. B-cell-depletion therapy over years for SSC in remission might represent a risk factor for infections. Additionally, the parenteral administration is associated with an increase in health care and patients are bound to regular hospital visits, which is a limiting factor for life quality.ObjectivesWe aimed to investigate if subcutaneous Abatacept could be used as a maintenance treatment in SSc-patients reaching a stable disease activity after a treatment with RTX administered according to our Grazer protocol over several years.MethodsIn this retrospective analysis, we retrieved data from 20 patients who fulfilled the diagnosis criteria for systemic sclerosis (SSc) according to the European League Against Rheumatism Scleroderma Trial and Research standards, who started a treatment with abatacept (ABA) after years of 500 mg RTX therapy every three months +/- MMF (Grazer protocol). The following clinical parameter were evaluated: modified Rodnan Skin Score (mRSS), Systemic Sclerosis Activity Score (SScAS), Systemic Sclerosis Severity Score (SScSS) and lung diffusing capacity for carbon monoxide (DLCO). Lab parameters like IgG, ANA, ENA and inflammation parameters were routinely assessed. Clinical data from baseline visit (BSL) (before ABA treatment start) and follow-up visit (FU) (after 6 months of treatment) were collected.ResultsWe included 20 SSc patients in this retrospective analysis who changed from RTX to Abatacept. The majority were female (n=16; 94.1%), with a mean age ±SD of 54.8 years ±11 and an average disease duration of 7.7 years ±4.5. In 17.9% (n=4) treatment needed to be stopped due to disease flare after three months (lung n=2, skin and tendons n=2).However, interestingly, Abatacept further decreased significantly mRSS between baseline visit and follow-up visit regarding the affected skin of the fingers. Thus, the mRSS went from 3.9±3.4 to 2.5±1.8 (pConclusionAbatacept might be a feasible option as a maintenance therapy after intensive immunomodulation with RTX and may give our patients the possibility to further improve their quality of life.The fact that 4 patients experienced a relapse after switching to Abatacept warrants further studies to find prognostic factors.Disclosure of InterestsNone declared

Digitalisierung der Gewässerschau

Additive manufacturing, also commonly referred to as 3D printing, stands to transform sand casting with binder jetting technology that can create sand molds with unmatched geometric complexity. With printed sand molds, castings can be optimized with regard to the strength-versus-weight trade-off and structures such as periodic lattices are now available within molds that are not possible with traditional casting technology. However, an increase in design complexity invites more challenges in terms of understanding and managing both the thermodynamics and physics of the casting process. Simulations of castings are more important than ever, and empirical in situ sensor data are required to validate high fidelity computer modeling (e.g., MAGMASOFT®). One novel solution is to leverage the design freedom of CAD-based solid modeling to introduce unique mold features specifically for housing sensors (Internet of Things) within the mold to enable the collection of a diversity of data at manifold locations: temperature, pressure, moisture, gas chemistries, motion of the molds and internal cores (shifting or rotation), and magnetic field. This report describes a proof of concept in which unprecedented levels of process monitoring were integrated—both wirelessly and wired—at strategic locations throughout a printed mold and inside of internal cores. The collected data were used to validate the quality of a casting in situ as well as to provide feedback for optimizing the casting process, mold design, and simulations. A trade-off was explored between sensor survivability and disposability.

Surface finish is an integral part of casting quality specifications. Rough internal surface finishes on castings can cause the loss of efficiency for both fluids and high-velocity gasses, and such is the case for turbocharger and intake manifold components. The University of Northern Iowa has been investigating mold material characteristics that affect surface smoothness for castings. The research was conducted on aluminum castings but has applications and relevance in ferrous alloys that do not exhibit defects such as penetration or fused sand defects. The study investigates the influence of molding media characteristics such as sand fineness, material type, and refractory coating selection. The goal of the project was to accomplish investment casting surface finishes in sand cast parts.

On the way to the Channel Management 4.0 Auf dem Weg zum Kanalmanagement 4.0

Computer-based process modeling has long been used for simulating the casting of metals. The information from these simulations has allowed metalcasters to reduce the occurrence of some defects that result from the metal’s volumetric contraction or entrainment of gasses while filling of the mold cavity. The majority of the analysis of the casting process has been limited to the metallic casting itself rather than the mold that contains the metal. High-temperature characterization of molding materials has yielded an increased understanding of the condition of the mold and core during solidification. Properties such as thermal expansion, high-temperature reactions, and changes in surface viscosity and decomposition gasses have yielded an increased understanding of the casting process. By using actual high-temperature mechanical property data generated at the foundry, the accuracy of the solidification modeling is improved and the range of capabilities is increased. The use of applied programming interface code can extend the software’s capabilities and yield additional information on the condition of the mold before, during and after solidification. Although once limited in release, new versions of the process modeling software will include the capability to both optimize process variables and develop material-specific datasets that increase both the accuracy and extend the capabilities of the analysis. Using applied programming interface code, the range of capabilities of the process modeling software can be extended to include information on veining defects, final casting dimensions and decomposition gas generation that can result in porosity defects or changes to the thermal properties of the mold.

Three-dimensional printing of sand molds and cores is changing the way castings are produced. During the last 5 years major advances in equipment have allowed metalcasters to realize casting designs at faster speeds than ever before. The elimination of tooling for mold and cores assemblies has allowed the industry new flexibility in design optimization, reduced labor, increased dimensional accuracy, and eliminated some defects associated with core assembly. While the equipment has been advancing, the materials used for printing have been very limited. The University of Northern Iowa has conducted new research into increasing the number of materials available for printing. These materials include regionally available resins, aggregates, and additives. These new materials have the ability to reduce the cost of printing and increase the number of applications while improving the casting quality. Veining defects in heavy iron or steel castings once prevalent on printed sand molds can be eliminated with engineered sand additives. Solidification rate and penetration defects can be addressed with specialty molding aggregates. This research has allowed the users of printed sand molds and cores to realize the potential of their sand printers and improve the quality of the castings they produced. The paper will detail the experiences with regionally available materials and compare their properties and performance with conventionally supplied materials.

Elucidating the influence of social context on drug reward is critical for understanding substance use disorders. Adolescents demonstrate enhanced sensitivity to drug and social rewards. However, the extent to which methamphetamine interacts with social reward in adolescents has not been thoroughly examined. Therefore, the present study used the conditioned place preference (CPP) model to examine the relationship between methamphetamine and social rewards in adolescent male rats. Sprague-Dawley rats (PND 30) were randomly assigned to one of the following four conditioning groups: saline alone (SA), methamphetamine alone (MA), saline with a social partner (SS) or methamphetamine with a social partner (MS). Testing occurred in a two-chamber biased apparatus across seven consecutive days using parameters presumed to be sub-threshold for establishing social- and methamphetamine-induced CPP. Similar to previous reports for nicotine and cocaine, the present results indicate that rats receiving methamphetamine with a social partner (i.e., MS) during conditioning demonstrated a significantly greater preference shift compared to all other groups. These findings further highlight the importance of social context in influencing the magnitude of drug reward during adolescence.

Background Rheumatoid arthritis is an immune-mediated disease, in which immune cell activation leads to destructive inflammation of the joints. In this context the treatment with JAK inhibitor tofacitinib has been proven to be effective. In mice treatment with tofacitinib resulted in reduced specific antibody responses, failure to generate germinal centres, and partial block of B cell development in the bone marrow. Conversely in vivo treatment of psoriasis arthritis patients as well as of rheumatioid arthritis patients with tofacitinib results in an increase of relative and absolute numbers of B cells in the first 4–8 weeks from beginning of treatment. It is known that mouse early B cell development is strongly dependent on IL-7 signalling, while this is not essential in humans. Nevertheless other cytokines are important in determining the fate and development of B lymphocytes. Hence, the outcome of JAK inhibition in early B cell development remains to be studied Objectives To assess the impact of JAK inhibition on early B cell development in vitro Methods We used a in vitro model in which CD34+ cells isolated from cord blood are cultivated subsequently in SCF, Flt3-L and IL-6, then SCF, Flt3-L and IL-7 and finally in cytokine-free medium(.1 The culture reproduces all stages of development from common lymphocytes progenitors to immature B cells. Results With the addition of tofacitinib to the in vitro culture we observed an increase in the absolute numbers of lymphoid precursors developing in vitro especially at week 5 and 6 of culture. Specifically, JAK inhibition led to an increase of pre-B and immature B cells by week 5 and 6. These data are in contrast with the early B cell development block observed in the tofacitinib treated mouse, but are in line with the rapid increase of B cells in peripheral blood after 4–8 weeks of tofacitinib treatment in patients. Analysis of induction of fate determining genes (EBF, E2A, PAX-5) showed an earlier and stronger induction of fate determining genes. Conclusions Our data indicate that JAK inhibition may promote early B cell development by enhancing the commitment of lymphoid precursors to the B cell compartment, contributing to a temporary increase in relative and absolute numbers of B cell in peripheral blood of treated patients. These data contribute to our understanding of human B cell development, prompt us to further analyse the quality of B cell output from the bone marrow in JAK inhibited patients, and may provide cues to understand the outcome of JAK inhibition treatment in rheumatic diseases. Reference [1] Kraus H, et al. A feeder-free differentiation system identifies autonomously proliferating B cell precursors in human bone marrow. J Immunol. 2014Feb 1;192(3):1044–54. Disclosure of Interest None declared

Background CTLA-4 modulates T-cell activation by inducing inhibitory signalling in T-cells. To date the direct effect of CTLA-4-Ig (abatacept) on B-cells has not been studied in detail. We hypothesise that CTLA-4 modulates B-cell function, B-cell homeostasis and B-cell receptor signalling in a T-cell-dependent and T-cell-independent manner. Objectives To assess the impact of abatacept on 1. B-cell activation and function in vitro; 2. B-cell signalling in vitro; 3 maintenance and specificity of memory B-cells specific for citrullinated protein antigens in vivo. Methods The time kinetic of CD80 and CD86 expression was measured by flow cytometry on isolated B-cells after stimulation via CD40, IL-21R, TLR9, and BCR in presence and absence of CTLA-4-Ig. Human B-cells lines (BJAB) deficient either for CD80 or CD86 were created using the CRISPR Cas9 technology and the effect of abatacept on CD80 respectively CD86 expression was assessed in these K.O. cell lines. CD80 and CD86 internalisation upon abatacept binding was studied by fluorescence microscopy. The effect of abatacept on PI3K-, NF-kB, PLCy-signalling and Ca-flux in B-cells was assessed. The effect of CTLA-4-Ig in vivo on the pool of memory cells specific for anti-citrullinated protein antigens was studied by limiting dilution assays performed before and during CTLA-4-Ig treatment. Results The expression of CD80 and CD86 on isolated human B-cells peaked at day 2–3 of culture after stimulation with CD40-L and IL-21. CTLA-4-Ig led to a decrease in CD80 and CD86 expression. These results were confirmed in human B-cell lines deficient either for CD80 or CD86. By fluorescence microscopy we were able to demonstrate, that abatacept treatment of cultured B-cells leads to a reduction in CD80 and CD86 expression mediated by a mechanism that involves internalisation of the receptors and bound CTLA-4-Ig. To test if CTLA-4-Ig binding activates signalling pathways, we stimulated B-cells with CTLA4-Ig or via BCR or CD40. We detected phosphorylation of p65 indicating activation of the canonical NF-kB pathway. The effect of CTLA-Ig in vivo on the pool of memory cells specific for autoantigens (anti-citrullinated protein antigens) was studied by limiting dilution assays in patients with rheumatoid arthritis. This assay is based on polyclonal stimulation of memory B-cells that leads to blast formation and antibody secretion in the supernatant. We performed limiting dilution experiments before and after start of CTLA-4-Ig treatment and observed a significant reduction of the absolute frequency of citrullinated antigen specific memory B-cells in response to treatment. Conclusions CTLA-4-Ig-treatment leads to a decrease in B-cells’ CD80 and CD86 expression, mediated by a mechanism that involves internalisation of the receptors and bound CTLA-4-Ig. CTLA-4-Ig activates the canonical NF-kB pathway in B-cells. In vivo CTLA-4-Ig treatment reduces the frequency of citrullinated antigen specific memory B-cells in patients with rheumatoid arthritis. Disclosure of Interest J. Thiel Grant/research support from: Research Grant from Bristol Myers Squibb, R. Lorenzetti: None declared, I. Janovska: None declared, C. Smulski: None declared, L. Walter: None declared, J. Staniek: None declared, T. Schleyer: None declared, R. Voll: None declared, N. Venhoff: None declared, M. Rizzi Grant/research support from: Research Grant from Bristol Myers Squibb

Background Some rheumatologic disorders (RD) may initially manifest with central nervous system (CNS) affection, mimicking the clinical, magnetic resonance imaging, and cerebrospinal fluid (CSF) findings of multiple sclerosis (MS). Vice versa MS might be difficult to separate from some RD because of the presence of autoantibodies (e.g. ANA) in up to 50%. The MRZ reaction (MRZR), composed of the three respective antibody indices (AI) against measles, rubella, and varicella zoster virus, has been found positive frequently in MS patients. However, it is unclear whether the MRZR is helpful to distinguish rheumatologic disorders with CNS involvement (RDwCNS) from MS. Objectives To investigate the MRZ reaction as a diagnostic tool to distinguish patients with RDwCNS from patients with MS. Methods The MRZR was evaluated in 35 patients with RDwCNS and compared to 70 sex- and age- matched MS patients. An AI result ≥1.5 was indicative for intrathecal IgG production against the respective pathogen. Two previously established stringency levels, MRZR-1 (≥1 of 3 AIs positive) and MRZR-2 (≥2 of 3 AIs positive), were applied. CNS involvement of RDwCNS was defined as clinical manifestation with neurological symptoms and signs of inflammation in CSF analysis and/or cerebral/spinal magnetic resonance imaging (MRI). MRZR results were compared using the Fisher’s exact test with p Results Within the RDwCNS group, 31 patients suffered from systemic lupus erythematosus, four had a small vessel vasculitis. In both groups 77.1% were female, mean age (±SD) was 43.2 years (±18.7) in RDwCNS and 47.5 years (±7.8) in MS (p=n .s.). All RDwCNS patients showed clinical symptoms indicative for CNS involvement and signs of inflammation in CSF analyses and/or MRI of the brain. In 52 MS patients autoantibody screening was performed. 42% were positive for ANA (n=20) or ANCA (n=2) in indirect immunofluorescence. Only 14.3% of RDwCNS patients had a positive MRZR-1 compared to 85.7% within the MS group (p 2.0 for a positive AI, the prevalence of positive MRZR-2 dropped to 5.7% (n=2) in the RDwCNS group compared to 54.3% (n=38) in the MS group (p Conclusions Considering the high specificity of the MRZR-2 for MS confirmed in this study, this laboratory test may be a helpful diagnostic tool to distinguish RDwCNS from MS. Disclosure of Interest None declared

The topic of the examinations deals with the numeric methods and a method comparison of the ground water modeling of the infiltration of surface water. The preservation of the ground water balance sheets in connection with the required diminution of the drain crowds upper two-dimensionally drainage represents a major theme in all industrial nations. For this reason the infiltration by surface water is legally stipulated in Germany in different federal states. At the example of the most simple form of a hollow infiltration two mathematical or numerical models are introduced and their efficiency is compared. This is the set of rules ATV and the RICHARDS method. The comparison shows the efficiency of the hollow infiltration which is partly confirmed in insitu measurements. The causes of the differences are partly also in attemps in the model attempts as these don’t appear in nature, however. Therefore it gets required to adapt the models in their form. Horizontal flow processes should be included also. The permeability coefficients kv and kh permeability of the soil are to use enriched liquid under consideration of the infiltration ones by air in form of the rain water far.

Purpose: To demonstrate a case of isolated pauci-immune vasculitis of the pituitary gland in which the diagnosis was attained by stereotactic biopsy. Methods: We report a 36-year-old man who presented with diabetes insipidus and secondary hypogonadism. Cranial MRI revealed an enhancing lesion of the pituitary stalk. A histological sample was obtained by stereotactic biopsy and pathological work-up provided the diagnosis of isolated pauci-immune vasculitis. No further organ manifestations or serologic signs of systemic vasculitis were identified. There was no response to immunosuppressive therapy and the patient refused treatment with Rituximab. Conclusions: Vasculitis of the pituitary gland is a rare condition. Less than 50 cases have been described to date in patients with granulomatosis with polyangiitis (GPA, or Wegener’s granulomatosis) and few case reports exist on pituitary involvement in other systemic vasculitis like Behcet’s disease and Cogan’s syndrome 1 2 3 4 5 6 7 8 9 . Pituitary involvement in GPA predominantly affects the posterior pituitary gland resulting in central diabetes insipidus (DI) but global or partial anterior pituitary dysfunction and compression of the optic chiasm has also been described (10). Relapse and permanent residual pituitary insufficiency is common despite immunosuppressive therapy 7 . Vasculitis of the pituitary gland without manifestation in other organs has been described in few patients and diagnosis of GPA was attained by the presence of proteinase-3 specific ANCA and/or transsphenoidal biopsy in these cases 10 11 12 . The case presented here demonstrates that pauci-immune vasculitis of the pituitary can occur in the absence of systemic disease. The diagnosis can be made by stereotactic biopsy but therapy remains a challenge.