Redox environment in stem and differentiated cells: A quantitative approach.

Stem cells are believed to maintain a specific intracellular redox status through a combination of enhanced removal capacity and limited production of ROS. In the present study, we challenge this assumption by developing a quantitative approach for the analysis of the pro- and antioxidant ability of human embryonic stem cells in comparison with their differentiated descendants, as well as adult stem and non-stem cells. Our measurements showed that embryonic stem cells are characterized by low ROS level, low rate of extracellular hydrogen peroxide removal and low threshold for peroxide-induced cytotoxicity.

[CELLS FORM AND THEIR SENSITIVITY TO LYTIC ACTIVITY OF NATURAL KILLER CELLS UNDER THE ANTIOXIDANT ACTION].

The present paper is an attempt to estimate the influence of cell surface morphology changes to functional activity under the effect of antioxidant, N-acetylcysteine (NAC), and alpha-lipoic asid (ALA). Two experimental parameters were used to characterize transformed fibroblasts 3T3-SV40 status. The functional one was the cell sensitivity to lysis by natural killer (NK) mouse splenocytes, and morphology index (cell form index) was a cell area.

Dihydrolipoic but not alpha-lipoic acid affects susceptibility of eukaryotic cells to bacterial invasion.

Sensitivity of eukaryotic cells to facultative pathogens can depend on physiological state of host cells. Previously we have shown that pretreatment of HeLa cells with N-acetylcysteine (NAC) makes the cells 2-3-fold more sensitive to invasion by the wild-type Serratia grimesii and recombinant Escherichia coli expressing gene of actin-specific metalloprotease grimelysin [1]. To evaluate the impact of chemically different antioxidants, in the present work we studied the effects of α-Lipoic acid (LA) and dihydrolipoic acid (DHLA) on efficiency of S.

Telomere shortening is a sole mechanism of aging in mammals.

We adduce proof that telomere shortening is the sole mechanism of aging. All apparent contradictions, particularly the absence of an inverse correlation between residual telomere length and donor age, are explained within the bounds of telomere theory. We explain in what way telomere shortening might be the cause of aging and lifespan restriction.

N-acetylcysteine increases susceptibility of HeLa cells to bacterial invasion.

Serratia grimesii are non-pathogenic bacteria capable, however, to invade eukaryotic cells provided that they synthesize intracellular metalloprotease grimelysin (Bozhokina et al. [2011] Cell. Biol.

N-acetylcysteine-induced changes in susceptibility of transformed eukaryotic cells to bacterial invasion.

The effect of N-acetylcysteine (NAC) on morphological and physiological properties of "normal" 3T3 and 3T3-SV40 fibroblasts was studied. Incubation of the cells with 10 and 20 mM NAC for 20 h resulted in a reversible increase in the intracellular level of reduced glutathione and disorganization of actin cytoskeleton. Surprisingly, upon removal of NAC, 3T3-SV40 fibroblasts demonstrated formation of well-adhered cells with structured 3T3-like stress-fibers.

Different metabolic effects of ganglioside GM1 in brain synaptosomes and phagocytic cells.

The metabolic effects of ganglioside GM1 were found to be quite different in brain synaptosomes and phagocytic cells. Incubation of rat brain cortex synaptosomes with GM1 was shown to decrease the production of reactive oxygen species induced by Fe2+-H2O2 system and measured by chemiluminometric method in the presence of luminol. Gangliosides GM1, GD1a, and GT1b significantly diminished the induced accumulation of lipid peroxidation product in brain synaptosomes, but protein kinase inhibitor (polymyxin B) abolished this effect.

The role of reactive oxygen species in membrane potential changes in macrophages and astrocytes.

Involvement of reactive oxygen species (ROS) in changes of the plasma membrane potential of mouse peritoneal macrophages and astrocytes (U118 cell line) under the action of different agents has been studied. Membrane potential was measured using the voltage-dependent fluorescent oxonol dye DiBAC4(3). Agonists which stimulate macrophages to release ROS (the fMLP peptide and platelet activating factor) caused prolonged hyperpolarization.

Quantitative studies on the respiratory burst generated in peritoneal macrophages.

The luminol-dependent chemiluminescence of macrophages during the zymosan-stimulated respiratory burst has been studied both in the absence and in the presence of the radical inhibitor 3,5 di-tert-butyl-4-hydroxyphenyl propionic acid. In addition, the consumption of luminol and of the inhibitor has been followed analytically. Based on the rates of the consumption of the inhibitor, an iteration procedure yields a value of 2.

Cell-cycle-dependent changes in membrane potential of L-929 cells caused by the effect of hydrogen peroxide.

The changes in membrane potential of L-929 fibroblasts caused by H2O2 (10-50 microM) at different phases of the cell cycle were investigated. The membrane potential of cells in G0, early G1, and G2/M responded to H2O2 by hyperpolarizing, while cells in late G1/S responded by depolarizing. Quinidine (50 microM), a blocker of Ca2+-dependent K+ conductance, inhibited the hyperpolarization response of L-929 cells to H2O2 in all cases.

Roles of reactive oxygen species: signaling and regulation of cellular functions.

Reactive oxygen species (ROS) are the side products (H2O2, O2.-, and OH.) of general metabolism and are also produced specifically by the NADPH oxidase system in most cell types.

Superoxide release is involved in membrane potential changes in mouse peritoneal macrophages.

Participation of reactive oxygen species (ROS) in the changes in macrophage membrane potential resulted from effects of different agonists has been studied. Treatment of macrophages with chemotactic peptide fMLP or platelet-activating factor (PAF) caused a brief depolarization followed by a long-lasting hyperpolarization. Lipopolysaccharide and interferon-gamma only depolarized the plasma membrane.

Hydrogen peroxide-induced chemotaxis of mouse peritoneal neutrophils.

Directed locomotion of mouse peritoneal neutrophils under agarose was studied, and activity of hydrogen peroxide (H2O2) as a chemoattractant was tested in its concentration range of 10(-6) to 10(-3) M. It has been found that H2O2 at low concentrations (about 10 microM) induces chemotactic activity. This activity was not affected by the presence of serum in the agarose medium.

Activation of murine macrophages by hydrogen peroxide.

Hydrogen peroxide at concentrations from 0.1 to 20 microM enhances phagocytosis and oxidative burst of murine peritoneal macrophages. The activation of these macrophage functions is paralleled by prolonged hyperpolarization and a transient increase in cytoplasmic free calcium concentration.