Automatic processing system for shadowgraph and interference patterns

The design and operation of an automatic system for the processing of shadowgraph and interference images are described. The system includes a two-coordinate processing table with an optical system for the projection of transparent images onto the photodetector, an image filter in the photodetector field, and a device for controlling the movement of the table and transmitting information to the minicomputer

A Tool for Computation of Changes in Na+, K+, Cl− Channels and Transporters Due to Apoptosis by Data on Cell Ion and Water Content Alteration

Monovalent ions are involved in a vast array of cellular processes. Their movement across the cell membrane is regulated by numerous channels and transporters. Identification of the pathways responsible for redistribution of ions and cell water in living cells is hampered by their strong interdependence. This difficulty can be overcome by computational analysis of the whole cell flux balance. Our previous computational studies were concerned with monovalent ion fluxes in cells under the conditions of balanced ion distribution or during transition processes after stopping the Na+/K+ pump. Here we analyze a more complex case—redistribution of ions during cell apoptosis when the parameters keep changing during the process. New experimental data for staurosporine-induced apoptosis of human lymphoma cells U937 have been obtained: the time course of changes in cellular K+, Na+, Cl−, and water content, as well as Rb+ fluxes as a marker of the Na/K pump activity. Using a newly developed computational tool, we found that alteration of ion and water balance was associated with a 55% decrease in the Na+/K+-ATPase rate coefficient over a 4-h period, with a time-dependent increase in potassium channel permeability, and a decrease in sodium channel permeability. The early decrease in [Cl−]i and cell volume were associated with an ~5-fold increase in chloride channel permeability. The developed approach and the presented executable file can be used to identify the channels and transporters responsible for alterations of cell ion and water balance not only during apoptosis but in other physiological scenarios

Metformin attenuates the effect of Staphylococcus aureus on airway tight junctions by increasing PKCζ-mediated phosphorylation of occludin.

Airway epithelial tight junction (TJ) proteins form a resistive barrier to the external environment, however, during respiratory bacterial infection TJs become disrupted compromising barrier function. This promotes glucose flux/accumulation into the lumen which acts as a nutrient source for bacterial growth. Metformin used for the treatment of diabetes increases transepithelial resistance (TEER) and partially prevents the effect of bacteria but the mechanisms of action are unclear. We investigated the effect of metformin and Staphylococcus aureus on TJ proteins, zonula occludins (ZO)-1 and occludin in human airway epithelial cells (H441). We also explored the role of AMP-activated protein kinase (AMPK) and PKCζ in metformin-induced effects. Pretreatment with metformin prevented the S. aureus-induced changes in ZO-1 and occludin. Metformin also promoted increased abundance of full length over smaller cleaved occludin proteins. The nonspecific PKC inhibitor staurosporine reduced TEER but did not prevent the effect of metformin indicating that the pathway may involve atypical PKC isoforms. Investigation of TJ reassembly after calcium depletion showed that metformin increased TEER more rapidly and promoted the abundance and localization of occludin at the TJ. These effects were inhibited by the AMPK inhibitor, compound C and the PKCζ pseudosubstrate inhibitor (PSI). Metformin increased phosphorylation of occludin and acetyl-coA-carboxylase but only the former was prevented by PSI. This study demonstrates that metformin improves TJ barrier function by promoting the abundance and assembly of full length occludin at the TJ and that this process involves phosphorylation of the protein via an AMPK-PKCζ pathway

On the effects of mechanical stress of biological membranes in modeling of swelling dynamics of biological systems

We highlight mechanical stretching and bending of membranes and the importance of membrane deformations in the analysis of swelling dynamics of biological systems, including cells and subcellular organelles. Membrane deformation upon swelling generates tensile stress and internal pressure, contributing to volume changes in biological systems. Therefore, in addition to physical (internal/external) and chemical factors, mechanical properties of the membranes should be considered in modeling analysis of cellular swelling. Here we describe an approach that considers mechanical properties of the membranes in the analysis of swelling dynamics of biological systems. This approach includes membrane bending and stretching deformations into the model, producing a more realistic description of swelling. We also discuss the effects of membrane stretching on swelling dynamics. We report that additional pressure generated by membrane bending is negligible, compared to pressures generated by membrane stretching, when both membrane surface area and volume are variable parameters. Note that bending deformations are reversible, while stretching deformation may be irreversible, leading to membrane disruption when they exceed a certain threshold level. Therefore, bending deformations need only be considered in reversible physiological swelling, whereas stretching deformations should also be considered in pathological irreversible swelling. Thus, the currently proposed approach may be used to develop a detailed biophysical model describing the transition from physiological to pathological swelling mode.National Aeronautics & Space Administration (NASA):80NSSC19M0049; PR Space Grant (NASA):NNX15AI11Hinfo:eu-repo/semantics/publishedVersio

Computation of Pump-Leak Flux Balance in Animal Cells

Background/Aims: Many vital processes in animal cells depend on monovalent ion transport across the plasma membrane via specific pathways. Their operation is described by a set of nonlinear and transcendental equations that cannot be solved analytically. Previous computations had been optimized for certain cell types and included parameters whose experimental determination can be challenging. Methods: We have developed a simpler and a more universal computational approach by using fewer kinetic parameters derived from the data related to cell balanced state. A file is provided for calculating unidirectional Na+, K+, and Cl- fluxes via all major pathways (i.e. the Na/K pump, Na+, K+, Cl- channels, and NKCC, KC and NC cotransporters) under a balanced state and during transient processes. Results: The data on the Na+, K+, and Cl- distribution and the pump flux of K+ (Rb+) are obtained on U937 cells before and after inhibiting the pump with ouabain. There was a good match between the results of calculations and the experimentally measured dynamics of ion redistribution caused by blocking the pump. Conclusion: The presented approach can serve as an effective tool for analyzing monovalent ion transport in the whole cell, determination of the rate coefficients for ion transfer via major pathways and studying their alteration under various conditions

Transition of the system to the balanced state as displayed in the file RESP.txt.

<p>Transition of the system to the balanced state as displayed in the file RESP.txt.</p