Metabolic Glycoengineering in hMSC-TERT as a Model for Skeletal Precursors by Using Modified Azide/Alkyne Monosaccharides

Metabolic glycoengineering enables a directed modification of cell surfaces by introducing target molecules to surface proteins displaying new features. Biochemical pathways involving glycans differ in dependence on the cell type; therefore, this technique should be tailored for the best results. We characterized metabolic glycoengineering in telomerase-immortalized human mesenchymal stromal cells (hMSC-TERT) as a model for primary hMSC, to investigate its applicability in TERT-modified cell lines. The metabolic incorporation of N-azidoacetylmannosamine (Ac₄ManNAz) and N-alkyneacetylmannosamine (Ac₄ManNAl) into the glycocalyx as a first step in the glycoengineering process revealed no adverse effects on cell viability or gene expression, and the in vitro multipotency (osteogenic and adipogenic differentiation potential) was maintained under these adapted culture conditions. In the second step, glycoengineered cells were modified with fluorescent dyes using Cu-mediated click chemistry. In these analyses, the two mannose derivatives showed superior incorporation efficiencies compared to glucose and galactose isomers. In time-dependent experiments, the incorporation of Ac₄ManNAz was detectable for up to six days while Ac₄ManNAl-derived metabolites were absent after two days. Taken together, these findings demonstrate the successful metabolic glycoengineering of immortalized hMSC resulting in transient cell surface modifications, and thus present a useful model to address different scientific questions regarding glycosylation processes in skeletal precursors.     

Damage microstructure evolution of helium ion irradiated SiC under fusion relevant temperatures

In-situ transmission electron microscopy (TEM) with ion irradiation has been used to study the damage microstructure evolution of He ion irradiated 4-H SiC at nuclear fusion relevant temperatures. The SiC samples were irradiated with 20?keV He ions at 25, 400, 800 and 1200?°C to a dose of 5.0 displacements per atom (DPA). At 25?°C, the material fully amorphises at 1.5 DPA and no He bubble nucleation occurs up to the doses studied. At 400 and 800?°C, He bubble nucleation occurs and the material remains crystalline. Bubble nucleation occurs at 2.0?DPA at 400?°C but occurs at only 0.5?DPA at 1200?°C. This is attributed the He atoms de-trapping from vacancies and migrating interstitially to larger He-vacancy clusters at higher temperatures, leading to faster nucleation of observable He bubbles. Helium platelets form at an irradiation temperature of 1200?°C at 0.5?DPA showing a preference for nucleation between the {0001} basal planes.     

Influence of mineral precipitation on the performance and long-term stability of Fe 0 -permeable reactive barriers: A review on the basis of 19 Fe 0 -reactive barrier sites

A comparative evaluation of the monitoring results from 19 Fe⁰-permeable reactive barriers (PRBs) is presented which considers changes of groundwater composition within the Fe⁰-PRBs, the type and distribution of mineral phases that precipitate, porosity reductions, extent of the load of inorganic groundwater constituents and the overall performance. The findings show that the changes in groundwater composition within the PRBs are extremely similar, independent of flow velocities, residence times, and concentrations of inorganic substances. In addition, the type and distribution of mineral precipitates do not show great variances with the largest mineral accumulations within the first 10 to 30 cm from the entrance face of the PRBs. From these comparisons it can be concluded that loads of dissolved inorganic species higher than 200 kg?·?a^–1?·?m^–2 limit the long-term performance of Fe⁰-PRBs up to a few years due to porosity losses caused by mineral precipitation, independent of the type of construction. Moreover, detailed calculations of porosity and hydraulic conductivity decreases of the PRBs at the Rheine and Tübingen sites show that the influence of mineral fouling on the hydraulic permeability of a PRB can only be estimated by accounting for the interaction between reduction in porosity, permeability and flow rate. The findings suggest that it is possible to pre-estimate the porosity and permeability development for a potential PRB.     

Investigating the Role of Conformational Effects on Laccase Stability and Hyperactivation under Stress Conditions

Fungal laccase from Steccherinum ochraceum 1833 displays remarkable stability under different harsh conditions: organic/buffer mixtures, thermal treatment, and microwave radiation. The behavior is particularly significant in the light of the sharp inactivation observed for two different fungal laccases. Laccase from S. ochraceum 1833 also displays hyperactivation under mild thermal treatment (60 °C). Molecular dynamics simulations at 80 °C explained how this laccase retains the geometry of the electron transfer pathway, thereby assuring electron transfer through the copper ions and thus maintaining its catalytic activity at high temperature. Spectroscopic studies revealed that the thermal activation corresponds to specific conformational changes in the protein. The results indicate that this laccase is potentially applicable under denaturing conditions that might be beneficial for the biotransformation of recalcitrant substrates.     

Achieving dynamic adaptation via management and interpretation of runtime models

In this article, we present a generic model-centric approach for realizing fine-grained dynamic adaptation in software systems by managing and interpreting graph-based models of software at runtime. We implemented this approach as the Graph-based Runtime Adaptation Framework (GRAF), which is particularly tailored to facilitate and simplify the process of evolving and adapting current software towards runtime adaptivity. As a proof of concept, we present case study results that show how to achieve runtime adaptivity with GRAF and sketch the framework's capabilities for facilitating the evolution of real-world applications towards self-adaptive software. The case studies also provide some details of the GRAF implementation and examine the usability and performance of the approach.