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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 149 (1988), S. 214-219 
    ISSN: 1432-072X
    Keywords: Protonmotive force ; Active transport ; RuMP-type methylotrophs ; δpH ; δΨ
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Formaldehyde accumulation ratios ([14CH2O]i/[14CH2O]o) as high as 12-fold were measured in anaerobic, CH3OH-energized, whole cell suspensions of the ribulose monophosphate (RuMP)-type methylotrophic strain T15. Uptake kinetics were extremely rapid, enabling the attainment of equilibrium in only 10–30 s. Transport appears to be energy-dependent and associated with the protonmotive force (pmf). Anaerobic incubation with 5 μM carbonyl p-(trifluoromethoxy)-phenylhydrazone (FCCP) led to 70%–90% reduction of the accumulation ratio. Though not as pronounced, diminished uptake was also observed in the presence of 140 μM nigericin, 161 μM valinomycin and 90 mM KSCN, commensurate with their effects on pmf. Accumulation of CH2O as a function of external pH followed a trend more similar to that of pmf than either δpH or δΨ. Preventing energization by incubation with 100 μM N,N′-dicyclohexylcarbodiimide (DCCD) led to nearly 80% inhibition of CH2O transport. Over short time periods it was possible to “chase” accumulated 14CH2O from previously loaded cells by collapsing pmf; however, this technique also indicated that significant 14CH2O incorporation began to occur within 3 min.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 55 (1997), S. 693-700 
    ISSN: 0006-3592
    Keywords: glucose ; lactate ; real-time determination ; hematopoietic cell culture ; colony-forming cells ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Glucose and lactate metabolic rates were evaluated for cultures of cord blood (CB) mononuclear cell (MNC), peripheral blood (PB) MNC, and PB CD34+ cell cultures carried out in spinner flasks and in T-flasks in both serum-containing and serum-free media. Specific glucose uptake rates (qgluc, in micromoles per cell per hour) and lactate generation rates (qlac) correlated with the percentage of colony-forming cells (CFC) present in the culture for a broad range of culture conditions. Specifically, the time of maximum CFC percentage in each culture coincided with the time of maximum qgluc and qlac in cultures with different seeding densities and cytokine combinations. A two-population model (Qlac = α[CFC] + β([TC] - [CFC]), where [TC] is total cell concentration; Qlac is volumetric lactate production rate in micromoles per milliliter per hour; α is qlac for an average CFC; and β is qlac for an average non-CFC) was developed to describe lactate production. The model described lactate production well for cultures carried out in both T-flasks and spinner flasks and inoculated with either PB or CB MNC or PB CD34+ cells. The values for α and β that were derived from the model varied with both the inoculum density and the cytokine combination. However, preliminary results indicate that cultures carried out under the same conditions from different samples with similar initial CD34+ cell content have similar values for β and β. These findings suggest that it should be possible to use lactate production data to predict the harvest time that corresponds to the maximum number of CFC in culture. The ability to harvest ex vivo hematopoietic cultures for transplantation when CFC are at a maximum has the potential to speed the rate at which immunocompromised patients recover. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 693-700, 1997.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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