A steady state model for analyzing the cellular binding, internalization and degradation of polypeptide ligands

We demonstrate that the interaction of polypeptide ligands with cells under physiological conditions can be described by a set of steady state equations. These equations include four new rate constants: V_r, the rate of insertion of receptors into the cell membrane; K_e, the endocytotic rate constant of occupied receptors; K_t, the turnover rate constant of unoccupied receptors; and K_h, the rate constant of hydrolysis of internalized ligand. Several simple procedures are described for determining these constants. In experiments in which epidermal growth factor and human fibroblasts were used, the cell-ligand interactions followed the predictions of the steady state model. The utility of the steady state equations is demonstrated by establishing the kinetic basis of the commonly observed “down regulation” phenomenon and by quantitating the effect of methylamine on the endocytotic and degradation rates of epidermal growth factor. We also show that the slope of a “Scatchard plot” of steady state binding data is a complex constant including terms for the endocytotic rate of both occupied and unoccupied receptors. The X-intercept of such a plot is a function of the insertion rate of new receptors, the internalization rate of occupied receptors and the degradation rate of the internalized ligand. The steady state equations allow one to predict changes in cellular ligand binding resulting from alterations in the four rate constants. They also provide a foundation for computer simulations of ligand-cell interactions, which closely correspond to experimental data. These approaches should facilitate studies on the control of cellular activities by these polypeptide ligands.     

On the mass distribution model for microbial cell populations

The full implications of a statistical model for growth of a microbial cell population using cell mass as the index of physiological state have been examined by solving the partial differential integral equations resulting from the model. Calculations reveal that a lag phase is predicted during the initial stages of batch growth although no specific cellular mechanism for the phenomenon of lag had been incorporated into the model. The model predicts several situations of batch and continuous growth in which the population density and biomass concentration show opposing trends due to significant variation in the cell mass distribution with time.     

Dynamic analysis of <Emphasis Type="Italic">Lactobacillus delbrueckii</Emphasis> subsp. <Emphasis Type="Italic">bulgaricus</Emphasis> CFL1 physiological characteristics during fermentation

This study aimed at examining and comparing the relevance of various methods in order to discriminate different cellular states of Lactobacillus bulgaricus CFL1 and to improve knowledge on the dynamics of the cellular physiological state during growth and acidification. By using four fluorescent probes combined with multiparametric flow cytometry, membrane integrity, intracellular esterase activity, cellular vitality, membrane depolarization, and intracellular pH were quantified throughout fermentations. Results were compared and correlated with measurements of cultivability, acidification activity (Cinac system), and cellular ability to recover growth in fresh medium (Bioscreen system). The Cinac system and flow cytometry were relevant to distinguish different physiological states throughout growth. Lb. bulgaricus cells maintained their high viability, energetic state, membrane potential, and pH gradient in the late stationary phase, despite the gradual decrease of both cultivability and acidification activity. Viability and membrane integrity were maintained during acidification, at the expense of their cultivability and acidification activity. Finally, this study demonstrated that the physiological state during fermentation was strongly affected by intracellular pH and the pH gradient. The critical pHi of Lb. bulgaricus CFL1 was found to be equal to pH 5.8. Through linear relationships between dpH and cultivability and pHi and acidification activity, pHi and dpH well described the time course of metabolic activity, cultivability, and viability in a single analysis.     

Stoffwechsel phenolischer Verbindungen in gesunden und Braunrost-infizierten Gersten- und Weizenvarietäten

Metabolism of phenolic compounds in healthy and brown rust-infected barley and wheat varieties
Infections with Puccinia isolates cause an accumulation of phenolic compounds both in resistant and susceptible isogenic lines of barley and wheat seedlings. The increased de novo biosynthesis of hydroxy-cinnamic and -benzoic acids is catalysed by the enzyme phenylalanine-ammonia-lyase (PAL) whose activity seems to be controlled not only by the substrate (phenylalanine) but also by the formed products (phenols). Depending on its actual physiological state every plant species shows as a reaction upon infection a typical behaviour. It is remarkable that healthy and rust-infected plants contain the same percentage of individual phenolic acids. An infection does not lead to an increase of individual hydroxycinnamic- or benzoic acids. There are only differences in quantity but no differences in quality. The pattern of phenolic acids seems to be largely genetically determined and formed according to the developmental stage of the plants. The hydroxycinnamic and -benzoic acids formed during the host-parasit-interactions seem to be, at least for the analysed combinations, more the consequence than the cause of resistance.     

Modellbank Biotechnologie – Ein Softwarepaket zur Modellauswahl,Optimierung und Simulation biotechnologischer Prozesse

The MODELBASE BIOTECHNOLOGY has been developed for gathering and processing data obtained from biochemical experiments. It covers the analysis of growth kinetics and production kinetics as well and may be used for calibration curve determination too. The implementation of models defined by the user is supported. Both, differential equations and their integrated forms are accepted. In addition to parameter estimation a statistical analysis can be carried on request. Two and three dimensional graphical representation facilitates the interpretation of results. Fields of application are biochemistry, biotechnology, microbiology, engineering, pharmacy, equipment design, education.