Approaches for molecular characterization of modified biopolymers

In this thesis, research on the molecular characterization of products obtained after structure modification of oligosaccharides, starch, model peptides, and bovine α-lactalbumin is described. The research goals comprised the development of analytical tools as well as the elucidation of molecular structures by using these tools. The structure of 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-oxidized gelatinized potato starch was analyzed after degrading it to fragments, using acid hydrolysis and methanolysis. Mass spectrometric analysis of the resulting oligomers revealed that the TEMPO oxidation proceeds in a block-wise manner. Also, a high acid-resistance of especially the α(1à4) glucuronic acid-glucuronic acid glycosidic linkage was observed, making oxidized starch a possible dietary fiber. The structure of epoxidized granular 1-allyloxy-2-hydroxypropyl-waxy maize starch was characterized by a similar approach using enzymatic hydrolysis. This showed that the created epoxy groups took part in subsequent reactions to form cross links and/or diols. Discrepancies in literature concerning the specificity of immobilized Candida antarctica lipase B in the acylation of oligosaccharides were explained. Molecular sieves, used to remove free water from the organic reaction medium, were found to be responsible for the catalysis of side-reactions. Next, hydroxy-aryl esters of various oligosaccharides were chemically produced. Via a peroxidase-mediated reaction, these esters could subsequently be coupled to the model peptide Gly-Tyr-Gly, as a proof-of-principle for enzymatic protein glycosylation. In addition, α-lactalbumin was glycated with various saccharides via the Maillard reaction. Products were studied using UPLC-ESI-TOF MS and size exclusion chromatography. A detailed view on the extent of glycation and the dispersity of the products was obtained. The glycation rate, extent of protein cross-linking, and the foam stability of the glycated α-lactalbumin depended on the type of saccharide used. Similar saccharide structures led to similar behavior. The glycation rate and the extent of protein cross-linking decreased when the degree of polymerization of the saccharide increased. Dehydrated Amadori products, as identified by UPLC ESI-TOF MS, were found to be an indicator of the formation of cross-linked protein. </p

Isolation of a suppressible nonsense mutant in mammalian cells

Journal ArticleAn HGPRT- cell line derived from mouse L cells has been shown to have the following properties: it is CRM'; the defective HGPRT molecules are altered in the carboxyterminal peptide; the mutant cells regain HGPRT activity when ochre-suppressor tRNA is microinjected into them, but not when amber-suppressor or wild-type tRNAs are injected. We conclude from these properties that this mutant cell line contains an ochre nonsense mutation (UAA) in the structural gene for HGPRT

Meteorological application of Apollo photography Final report

Development of meteorological information and parameters based on cloud photographs taken during Apollo 9 fligh

Evaluation of radiography as a screening method for detection and characterisation of congenital vertebral malformations in dogs

Congenital vertebral malformations (CVM) are common in brachycephalic ‘screw-tailed’ dogs; they can be associated with neurological deficits and a genetic predisposition has been suggested. The purpose of this study was to evaluate radiography as a screening method for congenital thoracic vertebral malformations in brachycephalic ‘screw-tailed’ dogs by comparing it with CT. Forty-nine dogs that had both radiographic and CT evaluations of the thoracic vertebral column were included. Three observers retrospectively reviewed the images independently to detect CVMs. When identified, they were classified according to a previously published radiographic classification scheme. A CT consensus was then reached. All observers identified significantly more affected vertebrae when evaluating orthogonal radiographic views compared with lateral views alone; and more affected vertebrae with the CT consensus compared with orthogonal radiographic views. Given the high number of CVMs per dog, the number of dogs classified as being CVM free was not significantly different between CT and radiography. Significantly more midline closure defects were also identified with CT compared with radiography. Malformations classified as symmetrical or ventral hypoplasias on radiography were frequently classified as ventral and medial aplasias on CT images. Our results support that CT is better than radiography for the classification of CVMs and this will be important when further evidence of which are the most clinically relevant CVMs is identified. These findings are of particular importance for designing screening schemes of CVMs that could help selective breeding programmes based on phenotype and future studies

Gaussian noise and time-reversal symmetry in non-equilibrium Langevin models

We show that in driven systems the Gaussian nature of the fluctuating force and time-reversibility are equivalent properties. This result together with the potential condition of the external force drastically restricts the form of the probability distribution function, which can be shown to satisfy time-independent relations. We have corroborated this feature by explicitly analyzing a model for the stretching of a polymer and a model for a suspension of non-interacting Brownian particles in steady flow.Comment: 6 pages, submitted to PR