Effizienz der Gentherapie mit adenoviralen Vektoren bei Ornithintranscarbamylse defizienten Mäusen in Abhängigkeit von OTC-Leadersequenz

Die Effizienz einer somatischen Gentherapie des Ornithintranscarbamylase-Mangels mit adenoviralen Vektoren wird am Modell der spfash-Maus untersucht. Spfash-Mäuse werden mit Vektoren transfiziert, die das komplette OTC-Transgen der Maus, des Menschen oder Hybride mit vertauschten Leadersequenzen enthalten. Auf ultradünnen Gefrierschnitten des Lebergewebes von Kontrollmäusen, unbehandelten und transfizierten spfash-Mäusen wird die intrazelluläre Verteilung von OTC, CPS I und ATPase(c) immunelektronenmikroskopisch dargestellt und morphometrisch quantifiziert. Die Ergebnisse dieser In-vivo-Studie zeigen, dass die transgene OTC sehr effizient in die Mitochondrien von spfash-Hepatozyten importiert wird und die Voraussetzung für eine metabolische Korrektur des OTC-Mangels gegeben ist. Die mitochondriale Importkapazität der transgenen OTC und damit der gentherapeutische Erfolg hängen jedoch entscheidend von spezies-spezifischen Unterschieden in der Leadersequenz ab. The efficiency of somatic gene therapy of OTC deficiency with adenoviral vectors is examined in the spfash mouse model. Spfash mice are transfected with vectors containing the complete murine OTC transgene, the complete human OTC transgene or hybrids with exchanged leadersequences. On ultrathin frozen sections of liver tissue of control mice, untreated and transfected spfash mice the intracellular distribution of OTC, CPS I and ATPase(c) is shown by immune electronmicroscopy and is morphometrically quantified. The results of this in-vivo study reveal a very efficient import of transgenic OTC into mitochondria of spfash hepatocytes, so that the precondition for a metabolic correction of OTC deficiency is fulfilled. However, the capacity of mitochondrial import of transgenic OTC and, therefore, of gene therapeutic success strongly depend on species specific differences in the leadersequence

Effects of Exogenous Yeast and Bacteria on the Microbial Population Dynamics and Outcomes of Olive Fermentations.

In this study, we examined Sicilian-style green olive fermentations upon the addition of Saccharomyces cerevisiae UCDFST 09-448 and/or Pichia kudriazevii UCDFST09-427 or the lactic acid bacteria (LAB) Lactobacillus plantarum AJ11R and Leuconostoc pseudomesenteroides BGM3R. Olives containing S. cerevisiae UCDFST 09-448, a strain able to hydrolyze pectin, but not P. kudriazevii UCDFST 09-427, a nonpectinolytic strain, exhibited excessive tissue damage within 4 weeks. DNA sequencing of fungal internal transcribed spacer (ITS) regions and comparisons to a yeast-specific ITS sequence database remarkably showed that neither S. cerevisiae UCDFST 09-448 nor P. kudriazevii UCDFST 09-427 resulted in significant changes to yeast species diversity. Instead, Candida boidinii constituted the majority (>90%) of the total yeast present, independent of whether S. cerevisiae or P. kudriazevii was added. By comparison, Lactobacillus species were enriched in olives inoculated with potential starter LAB L. plantarum AJ11R and L. pseudomesenteroides BGM3R according to community 16S rRNA gene sequence analysis. The bacterial diversity of those olives was significantly reduced and resembled control fermentations incubated for a longer period of time. Importantly, microbial populations were highly dynamic at the strain level, as indicated by the large variations in AJ11R and BGM3R cell numbers over time and reductions in the numbers of yeast isolates expressing polygalacturonase activity. These findings show the distinct effects of exogenous spoilage and starter microbes on indigenous communities in plant-based food fermentations that result in very different impacts on product quality. IMPORTANCE Food fermentations are subject to tremendous selective pressures resulting in the growth and persistence of a limited number of bacterial and fungal taxa. Although these foods are vulnerable to spoilage by unintended contamination of certain microorganisms, or alternatively, can be improved by the deliberate addition of starter culture microbes that accelerate or beneficially modify product outcomes, the impact of either of those microbial additions on community dynamics within the fermentations is not well understood at strain-specific or global scales. Herein, we show how exogenous spoilage yeast or starter lactic acid bacteria confer very different effects on microbial numbers and diversity in olive fermentations. Introduced microbes have long-lasting consequences and result in changes that are apparent even when levels of those inoculants and their major enzymatic activities decline. This work has direct implications for understanding bacterial and fungal invasions of microbial habitats resulting in pivotal changes to community structure and function

Turbulence: introduction to theory and applications of turbulent flows

This book provides a general introduction to the topic of turbulent flows. Apart from classical topics in turbulence, attention is also paid to modern topics. After studying this work, the reader will have the basic knowledge to follow current topics on turbulence in scientific literature. The theory is illustrated with a number of examples of applications, such as closure models, numerical simulations and turbulent diffusion, and experimental findings. The work also contains a number of illustrative exercises