Retinal glycoprotein enrichment by concanavalin a enabled identification of novel membrane autoantigen synaptotagmin-1 in equine recurrent uveitis.

Complete knowledge of autoantigen spectra is crucial for understanding pathomechanisms of autoimmune diseases like equine recurrent uveitis (ERU), a spontaneous model for human autoimmune uveitis. While several ERU autoantigens were identified previously, no membrane protein was found so far. As there is a great overlap between glycoproteins and membrane proteins, the aim of this study was to test whether pre-enrichment of retinal glycoproteins by ConA affinity is an effective tool to detect autoantigen candidates among membrane proteins. In 1D Western blots, the glycoprotein preparation allowed detection of IgG reactions to low abundant proteins in sera of ERU patients. Synaptotagmin-1, a Ca2+-sensing protein in synaptic vesicles, was identified as autoantigen candidate from the pre-enriched glycoprotein fraction by mass spectrometry and was validated as a highly prevalent autoantigen by enzyme-linked immunosorbent assay. Analysis of Syt1 expression in retinas of ERU cases showed a downregulation in the majority of ERU affected retinas to 24%. Results pointed to a dysregulation of retinal neurotransmitter release in ERU. Identification of synaptotagmin-1, the first cell membrane associated autoantigen in this spontaneous autoimmune disease, demonstrated that examination of tissue fractions can lead to the discovery of previously undetected novel autoantigens. Further experiments will address its role in ERU pathology

Evaluation of phosphatidylserine-dependent antiprothrombin antibody testing for the diagnosis of antiphospholipid syndrome: results of an international multicentre study.

OBJECTIVE: A task force of scientists at the International Congress on Antiphospholipid Antibodies recognized that phosphatidylserine-dependent antiprothrombin antibodies (aPS/PT) might contribute to a better identification of antiphospholipid syndrome (APS). Accordingly, initial and replication retrospective, cross-sectional multicentre studies were conducted to ascertain the value of aPS/PT for APS diagnosis. METHODS: In the initial study (eight centres, seven countries), clinical/laboratory data were retrospectively collected. Serum/plasma samples were tested for IgG aPS/PT at Inova Diagnostics (Inova) using two ELISA kits. A replication study (five centres, five countries) was carried out afterwards. RESULTS: In the initial study (n = 247), a moderate agreement between the IgG aPS/PT Inova and MBL ELISA kits was observed (k = 0.598). IgG aPS/PT were more prevalent in APS patients (51%) than in those without (9%), OR 10.8, 95% CI (4.0-29.3), p < 0.0001. Sensitivity, specificity, positive (LR+) and negative (LR-) likelihood ratio of IgG aPS/PT for APS diagnosis were 51%, 91%, 5.9 and 0.5, respectively. In the replication study (n = 214), a moderate/substantial agreement between the IgG aPS/PT results obtained with both ELISA kits was observed (k = 0.630). IgG aPS/PT were more prevalent in APS patients (47%) than in those without (12%), OR 6.4, 95% CI (2.6-16), p < 0.0001. Sensitivity, specificity, LR + and LR- for APS diagnosis were 47%, 88%, 3.9 and 0.6, respectively. CONCLUSIONS: IgG aPS/PT detection is an easily performed laboratory parameter that might contribute to a better and more complete identification of patients with APS.info:eu-repo/semantics/publishedVersio

High temperature oxidation of EB-PVD TBCs on Pt-diffused single crystal Ni superalloy

The paper presents the results of high temperature oxidation of EB-PVD (Electron Beam Physical Vapor Deposition) TBCs (Thermal Barrier Coatings) deposited on a Pt-diffused second generation single crystal Ni-based superalloy. The Pt-diffused bond coating was obtained using a PVD (Physical Vapor Deposition) method applying a Closed Hollow Cathode (CHC) for the deposition of 5 µm of Pt layer and subsequent annealing at 1140 °C for 2 h in vacuum. The TBC system was thermally cycled in 1.25 h cycles at 1100 °C for 730 h (584 cycles) until delamination of 10% of Yttria Stabilized Zirconia (YSZ) top coating. A non-destructive 3D optical scanning method was applied for a macroscopic examination of the YSZ delamination which allowed predicting and revealing of the regions of YSZ buckling prior to its spallation. The microstructure of the TBC system in the as-deposited and thermally cycled conditions was studied using FEG-SEM. The growth and evolution of the Thermally Grown Oxide (TGO) scale was performed using high resolution FEG-S/TEM and Focused Ion Beam (FIB) methods. The TGO in the as-deposited condition was found to consist of porous a-Al2O3 which grew to form distinctive dense and columnar grains with ionic diffusion of reactive elements (Hf and Y) through grain boundaries during high temperature exposure. Additionally the formation of nanometric NiAl2O4 islands as well as high volume oxides containing Ni was found to accelerate the spallation of the YSZ top coat

Identification of autoantigens in body fluids by combining pull-downs and organic precipitations of intact immune complexes with quantitative label-free mass spectrometry.

Most autoimmune diseases are multifactorial diseases and are caused by the immunological reaction against a number of autoantigens. Key for understanding autoimmune pathologies is the knowledge of the targeted autoantigens, both initially and during disease progression. We present an approach for autoantigen identification based on isolation of intact autoantibody-antigen complexes from body fluids. After organic precipitation of high molecular weight proteins and free immunoglobulins, released autoantigens were identified by quantitative label-free liquid chromatography mass spectrometry. We confirmed feasibility of target enrichment and identification from highly complex body fluid proteomes by spiking of a pre-defined antigen-antibody complex at low level of abundance. As a proof of principle we studied the blinding disease autoimmune uveitis, which is caused by autoreactive T-cells attacking the inner eye and is accompanied by autoantibodies. We identified three novel autoantigens in the spontaneous animal model equine recurrent uveitis (secreted acidic phosphoprotein osteopontin, extracellular matrix protein 1 and metalloproteinase inhibitor 2) and confirmed presence of the corresponding autoantibodies in 15 - 25% of patient samples by enzyme-linked immunosorbent assay. Thus, this workflow led to the identification of novel autoantigens in autoimmune uveitis and may provide a versatile and useful tool to identify autoantigens in other autoimmune diseases in future

Interactive Learning of Inverse Kinematics with Null-space Constraints using Recurrent Neural Networks

Emmerich C, Nordmann A, Swadzba A, Wrede S, Steil JJ. Interactive Learning of Inverse Kinematics with Null-space Constraints using Recurrent Neural Networks. Presented at the International Conference on Cognitive Systems, Vienna

Assisted Gravity Compensation to Cope with the Complexity of Kinesthetic Teaching on Redundant Robots

Emmerich C, Nordmann A, Swadzba A, Steil JJ, Wrede S. Assisted Gravity Compensation to Cope with the Complexity of Kinesthetic Teaching on Redundant Robots. Presented at the International Conference on Robotics and Automation (ICRA), Karlsruhe.Facilitating efficient reconfiguration of advanced robot systems to new tasks or environments is an ongoing research challenge. While imitation learning methods as well as modeling and simulation tools are continuously improving to reduce the inherent complexity of reconfiguration, novel robot systems are entering the field which pose new challenges. This paper addresses one important challenge in this area, which is the programming of kinematically redundant robots. We show as a result of a field study with 48 industrial workers that standard programming-by-demonstration methods for teaching task-space trajectories on a redundant robot using physical human-robot interaction are too complex for non-expert human tutors. We therefore propose a new interaction concept for redundant robot systems, Assisted Gravity Compensation, based on a hierarchical control scheme, separating task-space programming from the redundancy resolution. The user is actively assisted by a given redundancy resolution while kinesthetically teaching task-space trajectories. The results of the field study show that by this means the complexity of a kinesthetic teaching task is reduced, which is revealed by an improved task performance and user experience, making kinesthetic teaching an efficient programming-by-demonstration method for redundant robots

Thermochromism and switchable paramagnetism of cobalt(ii) in thiocyanate ionic liquids

Temperature-dependent switching of paramagnetism of a cobalt(ii) complex is observed in an ionic liquid solution.</p