Ligand-Directed Chemistry on Glycoside Hydrolases – A Proof of Concept Study

Selective covalent labelling of enzymes using small molecule probes has advanced the scopes of protein profiling. The covalent bond formation to a specific target is the key step of activity-based protein profiling (ABPP), a method which has become an indispensable tool for measuring enzyme activity in complex matrices. With respect to carbohydrate processing enzymes, strategies for ABPP so far involve labelling the active site of the enzyme, which results in permanent loss of activity. Here, we report in a proof of concept study the use of ligand-directed chemistry (LDC) for labelling glycoside hydrolases near – but not in – the active site. During the labelling process, the competitive inhibitor is cleaved from the probe, departs the active site and the enzyme maintains its catalytic activity. To this end, we designed a building block synthetic concept for small molecule probes containing iminosugar-based reversible inhibitors for labelling of two model β-glucosidases. The results indicate that the LDC approach can be adaptable for covalent proximity labelling of glycoside hydrolases.T. M. W. thanks the FWF (Wien, Austria) for financial support (project number P30372-B21). Authors from TU Graz acknowledge support from NAWI Graz.Peer reviewe

Demographic, clinical and antibody characteristics of patients with digital ulcers in systemic sclerosis: data from the DUO Registry

OBJECTIVES: The Digital Ulcers Outcome (DUO) Registry was designed to describe the clinical and antibody characteristics, disease course and outcomes of patients with digital ulcers associated with systemic sclerosis (SSc). METHODS: The DUO Registry is a European, prospective, multicentre, observational, registry of SSc patients with ongoing digital ulcer disease, irrespective of treatment regimen. Data collected included demographics, SSc duration, SSc subset, internal organ manifestations, autoantibodies, previous and ongoing interventions and complications related to digital ulcers. RESULTS: Up to 19 November 2010 a total of 2439 patients had enrolled into the registry. Most were classified as either limited cutaneous SSc (lcSSc; 52.2%) or diffuse cutaneous SSc (dcSSc; 36.9%). Digital ulcers developed earlier in patients with dcSSc compared with lcSSc. Almost all patients (95.7%) tested positive for antinuclear antibodies, 45.2% for anti-scleroderma-70 and 43.6% for anticentromere antibodies (ACA). The first digital ulcer in the anti-scleroderma-70-positive patient cohort occurred approximately 5 years earlier than the ACA-positive patient group. CONCLUSIONS: This study provides data from a large cohort of SSc patients with a history of digital ulcers. The early occurrence and high frequency of digital ulcer complications are especially seen in patients with dcSSc and/or anti-scleroderma-70 antibodies

Digitaler Zwilling Straßenverkehr - Infrastrukturbasierte Unterstützung für kooperatives, vernetztes, automatisiertes Fahren

Am Beginn des Projekts DIGEST stand die Herausforderung, wie ein Straßenbetreiber in Zukunft mit dem zunehmenden Aufkommen von automatisierten Fahrzeugen – und der so steigenden Komplexität des Gesamtverkehrs – einen sicheren und effizienten Betrieb gewährleisten können. Die Kernfrage der Ausschreibung war dabei, wie ein digitaler Zwilling des Verkehrssystems Straße helfen kann. Das Projekt DIGEST wurde von einem internationalen Konsortium umgesetzt. Basierend auf einem breiten Stakeholder-Involvement zur Identifikation von Nutzeranforderungen wurden ein Informationsmodell (Basis: DIKW-Modell) und eine IT-Architektur (aufbauend auf OPC UA) erarbeitet. Dieses Konzept konnte mittels dreier Demonstratoren validiert werden. Abschließend wurden Betriebs- und Rollenmodelle erarbeitet und international diskutiert. In DIGEST konnte in unterschiedlichen Umgebungen (Autobahn, Stadt) demonstriert werden, dass mittels einheitlicher Informationsmodellierung und abgeleiteter IT-Architektur eine infrastrukturbasierte Unterstützungsleistung für automatisierte Fahrzeuge möglich ist. Das in DIGEST erarbeitete Konzept des digitalen Zwillings wird als CCAM Decision Support Platform bezeichnet. Damit kann automatisiertes Fahren durch vertrauenswürdige Infrastrukturinformationen sicherer gemacht werden. Die Rolle der D-A-CH-Region und ihrer Schlüssel-Stakeholder in diesem Themenfeld konnte weiter gefestigt und konsolidiert werden

Infrastructure-Based Digital Twins for Cooperative, Connected, Automated Driving and Smart Road Services

Driving requires continuous decision making from a driver taking into account all available relevant information. Automating driving tasks also automates the related decisions. However, humans are very good at dealing with bad quality, fuzzy, informal and incomplete information, whereas machines generally require solid quality information in a formalized format. Therefore, the development of automated driving functions relies on the availability of machine-usable information. A digital twin contains quality controlled information collected and augmented from different sources, ready to be supplied to such an automated driving function. An information model that describes all conceivably relevant information is necessary. To this end, a list of requirements that such an information model should meet is proposed and each requirement is argued for. Based on the anticipated services and applications that such a system should support, a collection of requirements for system architecture is derived. Information modeling is performed for selected relevant information groups. A system architecture has been proposed and validated with three different implementations, addressing several different applications to support decisions at a highway tunnel construction site in Austria and throughout the Test Bed Lower Saxony in Germany

N-Alkylated Iminosugar Based Ligands: Synthesis and Inhibition of Human Lysosomal β-Glucocerebrosidase

The scope of a series of N-alkylated iminosugar based inhibitors in the d-gluco as well as d-xylo configuration towards their interaction with human lysosomal β-glucocerebrosidase has been evaluated. A versatile synthetic toolbox has been developed for the synthesis of N-alkylated iminosugar scaffolds conjugated to a variety of terminal groups via a benzoic acid ester linker. The terminal groups such as nitrile, azide, alkyne, nonafluoro-tert-butyl and amino substituents enable follow-up chemistry as well as visualisation experiments. All compounds showed promising inhibitory properties as well as selectivities for β-glucosidases, some exhibiting activities in the low nanomolar range for β-glucocerebrosidase.Science, Faculty ofNon UBCChemistry, Department ofReviewedFacult

Aspects of structural order in Bi-209-containing particles for potential MRI contrast agents based on quadrupole enhanced relaxation

Quadrupole relaxation enhancement (QRE) has been suggested as the key mechanism for a novel class of field-selective, potentially responsive magnetic resonance imaging contrast agents. In previous publications, QRE has been confirmed for solid compounds containing Bi-209 as the quadrupolar nucleus (QN). For QRE to be effective in aqueous dispersions, several conditions must be met, i.e. high transition probability of the QN at the H-1 Larmor frequency, water exchange with the bulk and comparatively slow motion of the Bi-carrying particles. In this paper, the potential influence of structural order within the compounds (crystallinity') on QRE was studied by nuclear quadrupole resonance (NQR) spectroscopy in one crystalline and two amorphous preparations of Triphenylbismuth (BiPh3). The amorphous preparations comprised (1) a shock-frozen melt and (2) a granulate of polystyrene which contained homogeneously distributed BiPh3 after common dissolution in THF and subsequent evaporation of the solvent. In contrast to the crystalline powder which exhibits strong, narrow NQR peaks the amorphous preparations did not reveal any NQR signals above the noise floor. From these findings, we conclude that the amorphous state leads to a significant spectral peak broadening and that for efficient QRE in potential contrast agents structures with a high degree of order (near crystalline) are required