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Whatever the oxygenation condition, CaO2 was increased after rhEpo (Table 2) resulting in a higher maximal systemic O2 delivery both in normoxia (P = 0.06) and in hypoxia (P<0.05) (Fig. 6B).
C-labelling experiments were performed in at least two replicate cultures under each oxygenation condition.
After a warm-up period of 10 min at a low intensity (40 % Pmax), the subjects cycled for 30 min at 65%% Pmax in the respective oxygenation condition during training.
The extent of the differences induced by the drug treatment can be better highlighted by performing a PCA on each subset of NMR data limited to one cell line and one oxygenation condition.
The measured uptake and excretion rates and the rates of metabolic precursor depletion to biosynthesis, as determined from the composition of P. pastoris biomass previously reported for each oxygenation condition [ 18], were combined with a set of linearly independent equations obtained from METAFoR analysis to render the complete linear system solvable.
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In this comparative study, the gene expression profiles of S. cerevisiae and P. pastoris, a Crabtree positive and Crabtree negative yeast, respectively, were analyzed for three different oxygenation conditions (normoxic, oxygen-limited and hypoxic) under recombinant protein producing conditions in chemostat cultivations.
Completely respirative metabolism was observed in fully aerobic conditions and fully fermentative metabolism in anaerobic conditions and in the three different reduced oxygenation conditions the actual amount of oxygen available was observed to result in different flux contributions through respirative and fermentative pathways.
However, the fluxes of glucose 6-phosphate isomerase, fructose bisphosphate aldolase, 3-phosphoglycerate kinase, pyruvate decarboxylase, and alcohol dehydrogenase reactions were substantially lower under 2.8% oxygenation conditions than under even lower oxygen availability [ 23] whereas the corresponding transcript levels did not, as expected, show consistent behavior [ 21].
The appearance of the transporters in the up-regulated molecular paths suggests that in 0.5% oxygenation conditions the low availability of oxygen may have limited the generation of proton gradient across the mitochondrial membrane by the electron transfer chain of S. cerevisiae and, thus, the transport required reorganization.
We used three oxygenation conditions in our experiments: unregulated dissolved oxygen (DO), "Oxy-const", regulated DO, "Oxy-regul", and high DO, "Oxy-high" conditions.
Chemostat cultivations were performed with five different oxygenation conditions: 20.9%, 2.8%, 1.0%, 0.5% and 0.0% oxygen of the chemostat inlet gas.
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