An analysis of countercurrent exchange with emphasis on renal function

An analysis of countercurrent exchange in a U-tube is presented for a single-solute, constant-volume flow rate system with spatially varying source fluxes and permeabilities. Analytical solutions are given for the steady-state equations and numerical solutions for the unsteady-state equations. The solutions indicate that an external source of solute delivered to the stream flowing away from the U-tube bend can be distributed by the exchanger so that the concentration in both limbs increases toward the bend. In particular, there exist source fluxes whose magnitude decreases monotonically toward the bend for which the maximum solute concentration occurs at the bend. The point at which a concentration maximum occurs is governed principally by the solute permeability of the barrier separating the two limbs and by the volume flow rate through the exchanger. The system dynamics depend strongly on the relative cross-sectional areas of the two limbs or, equivalently, on the flow velocities within them. The model is used as a basis for discussion of various functional aspects of the renal vasa recta system.     

Countercurrent exchange in the inner renal medulla: Vasa recta-descending limb system

A model of countercurrent exchange has been developed to simulate transport of salt, urea and water among vasa recta and descending limbs of the loop of Henle in the inner medulla. These vessels are abstracted as three concentric cylinders: the inne one represents descending vasa recta, the middle one represents ascending vasa recta and the outer one represents descending limbs. The capillary plexus, which connects the ascending and descending vasa recta, is modeled as a series of well-mixe compartments. Multicomponent transport equations for the sytem are derived from steady state mass balances and simple passive flux relations. The resulting set of nonlinear equations are solved numerically by an iterative Gauss-Seidel algorithm with under-relaxation. Simulations yield the salt and urea concentrations as well as volume flow rates in all tubes and compartments. The simulations indicate that solute concentrations can increase monotonically toward the papillae even if all transport processes within the exchanger are passive and source fluxes decrease monotonically toward the papillae.     

Novel anti-prostate cancer scaffold identified by the combination of in silico and cell-based assays targeting the PI3K-AKT-mTOR pathway

Phenotypic assays were performed in prostate cancer cell lines to describe the biological activity of PI3K-AKT-mTOR pathway inhibitors retrieved from the virtual screening initiative. These novel chemicals share in common the aminopyridine scaffold, hitting PC-3 cells in macromolar range, with selectivity index over fibroblast cell lines. Moreover, a preliminary study of the mode of action by flow cytometry assay pointed out that these compounds had a rapamycin-like response for the PI3K-AKT-mTOR pathway modulation.     

Extracellular noise-induced stochastic synchronization in heterogeneous quorum sensing network

Quorum sensing is a bacterial mechanism used to synchronize the coordinated response of a microbial population. Because quorum sensing in Gram-negative bacteria depends on release and detection of a diffusible signaling molecule (autoinducer) among a multicellular group, it is considered a simple form of cell-cell communication for the purposes of mathematical analysis. Stochastic equation systems have provided a common approach to model biochemical or biophysical processes. Recently, the effect of noise to synchronize a specific homogeneous quorum sensing network was successfully modeled using a stochastic equation system with fixed parameters. The question remains of how to model quorum sensing networks in a general setting. To address this question, we first set a stochastic equation system as a general model for a heterogeneous quorum sensing network. Then, using two relevant biophysical characteristics of Gram-negative bacteria (the permeability of the cell membrane to the autoinducer and the symmetry of autoinducer diffusion) we construct the solution of the stochastic equation system at an abstract level. The solution indicates that stable synchronization of a quorum sensing network is robustly induced by an environment with a heterogenous distribution of extracellular and intracellular noise. The synchronization is independent of the initial state of the system and is solely the result of the connectivity of the cell network established through the effects of extracellular noise.     

Role of NADH/NAD+ transport activity and glycogen store on skeletal muscle energy metabolism during exercise: in silico studies

Skeletal muscle can maintain ATP concentration constant during the transition from rest to exercise, whereas metabolic reaction rates may increase substantially. Among the key regulatory factors of skeletal muscle energy metabolism during exercise, the dynamics of cytosolic and mitochondrial NADH and NAD+ have not been characterized. To quantify these regulatory factors, we have developed a physiologically based computational model of skeletal muscle energy metabolism. This model integrates transport and reaction fluxes in distinct capillary, cytosolic, and mitochondrial domains and investigates the roles of mitochondrial NADH/NAD+ transport (shuttling) activity and muscle glycogen concentration (stores) during moderate intensity exercise (60% maximal O2 consumption). The underlying hypothesis is that the cytosolic redox state (NADH/NAD+) is much more sensitive to a metabolic disturbance in contracting skeletal muscle than the mitochondrial redox state. This hypothesis was tested by simulating the dynamic metabolic responses of skeletal muscle to exercise while altering the transport rate of reducing equivalents (NADH and NAD+) between cytosol and mitochondria and muscle glycogen stores. Simulations with optimal parameter estimates showed good agreement with the available experimental data from muscle biopsies in human subjects. Compared with these simulations, a 20% increase (or approximately 20% decrease) in mitochondrial NADH/NAD+ shuttling activity led to an approximately 70% decrease (or approximately 3-fold increase) in cytosolic redox state and an approximately 35% decrease (or approximately 25% increase) in muscle lactate level. Doubling (or halving) muscle glycogen concentration resulted in an approximately 50% increase (or approximately 35% decrease) in cytosolic redox state and an approximately 30% increase (or approximately 25% decrease) in muscle lactate concentration. In both cases, changes in mitochondrial redox state were minimal. In conclusion, the model simulations of exercise response are consistent with the hypothesis that mitochondrial NADH/NAD+ shuttling activity and muscle glycogen stores affect primarily the cytosolic redox state. Furthermore, muscle lactate production is regulated primarily by the cytosolic redox state.     

Characterization of the Aerial Escape Response of the African Butterfly Fish, Pantodon buchholzi Peters

We studied the startle response of the African butterfly fish, Pantodon buchholzi (Osteoglossomorpha, Osteoglossoidea). It is an upward movement, mediated by abduction of the pectoral fins, and is elicited by mechanical and visual stimuli. Because this fish inhabits the first few centimeters beneath the water surface, its startle response results in an aerial excursion that may be described as ballistic-like, following a motion as defined by linear acceleration. We show that the aerial excursion is well-modeled by a parabola. On average, a fish jumps no more than twice its height and travels horizontally about five times its standard length. The fish may exhibit variable in-flight trunk and fin movements, but neither increases the travel distance in air following the initial in-water propulsive event. Similar vertical jumps also occur entirely within the water column suggesting that this motor behavior of Pantodon is a general escape behavior analogous to a Mauthner neuron-induced escape response. The variability in its posture in air and its direction of motion after reentering the water enhances this act of vertical flight as a step in this fish's escape behavior. The aerial aspect of its escape behavior is only a consequence of its position in the water column.     

System for dynamic measurements of membrane capacitance in intact epithelial monolayers.

Dynamic measurements of exocytosis have been difficult to perform in intact epithelial monolayers. We have designed a system that estimates with +/-1% accuracy (99% confidence) the total membrane capacitance of monolayers represented by a lumped model. This impedance measurement and analysis system operates through a conventional transepithelial electrophysiology clamp, performing all signal measurements as frequently as every 5 s. Total membrane capacitance (the series combination of apical and basolateral membranes) is the inverse of one of three unique coefficients that describe the monolayer impedance. These coefficients are estimated using a weighted, nonlinear, least-squares algorithm. Using the estimated coefficients, solution ranges for individual membrane parameters are calculated, frequently providing results within +/-20% of true values without additional electrophysiological measurements. We determined the measurement system specifications and statistical significance of estimated parameters using 1) analytical testing with circuit simulation software and equation-generated data; 2) a system noise analysis combined with Monte Carlo simulations; and 3) analog model circuits for calibration of the electronic system and to check equation-generated results. Finally, the time course of capacitance changes associated with purinergically stimulated mucin exocytosis are quantified in monolayers of the colonic goblet cell-like cell line HT29-CI.16E.