Role of Glycosylation in Conformational Stability, Activity, Macromolecular Interaction and Immunogenicity of Recombinant Human Factor VIII

Factor VIII (FVIII) is a multi-domain glycoprotein that is an essential cofactor in the blood coagulation cascade. Its deficiency or dysfunction causes hemophilia A, a bleeding disorder. Replacement using exogenous recombinant human factor VIII (rFVIII) is the first line of therapy for hemophilia A. The role of glycosylation on the activity, stability, protein–lipid interaction, and immunogenicity of FVIII is not known. In order to investigate the role of glycosylation, a deglycosylated form of FVIII was generated by enzymatic cleavage of carbohydrate chains. The biochemical properties of fully glycosylated and completely deglycosylated forms of rFVIII (degly rFVIII) were compared using enzyme-linked immunosorbent assay, size exclusion chromatography, and clotting activity studies. The biological activity of degly FVIII decreased in comparison to the fully glycosylated protein. The ability of degly rFVIII to interact with phosphatidylserine containing membranes was partly impaired. Data suggested that glycosylation significantly influences the stability and the biologically relevant macromolecular interactions of FVIII. The effect of glycosylation on immunogenicity was investigated in a murine model of hemophilia A. Studies showed that deletion of glycosylation did not increase immunogenicity

Surface-sensitive X-ray diffraction across the pressure gap

In this chapter surface-sensitive X-ray diffraction is introduced as an important crystallographic tool for the investigation of surfaces and nanostructures under high pressure reaction conditions and elevated temperatures which are relevant for industrial catalysis. After the introduction surface-sensitive X-ray diffraction methods are briefly explained and specialized instrumentation developed for the in situ investigation of surfaces and nanostructures across the pressure gap is presented combined with simultaneous measurement of the concentrations of reactants and products. In the following an overview of the experimental results is given: First the (near)-ambient pressure oxidation of 3d, 4d, and 5d transition metals is discussed which are relevant for oxidation catalysis. Afterwards catalytic reaction experiments in batch mode are reported, followed by an overview of current research using a flow reactor for surface-sensitive X-ray diffraction. Finally a perspective is given for future research directions

Who am I?:shaping young children’s identities through everyday narratives

Abstract Young children have largely been neglected in narrative identity research. This chapter is based on the premise that identity construction is a process that begins in the early years and is formed through the everyday narratives that comprise the daily interactions through which identities are constructed, negotiated, and performed. These everyday narratives provide children with the sense of who they are, who they will become, what kind of children they are expected to be, and the nature of the world in which they live. The chapter is based on research into everyday narratives produced in early education settings. I suggest that four themes frame the construction of children’s narrative identities: the possession of material items; skills and competences; peer relationships; and the child’s relation to the rules and conventions of the child care center