Effect of gold nanoparticles coated with plasma components on ADP-induced platelet aggregation.

We studied the effect of gold nanospheres coated with components of autologous blood plasma on ADP-induced platelet aggregation. Gold nanoparticles in the chosen concentration range (5-40 µM) and particle size (5-30 nm) with or without coating produced no activating effect on platelet aggregation caused by aggregation inductor ADP in all applied doses (1.6, 2.0, and 5.0 µM). Nanoparticles with a diameter of >60 nm inhibited platelet aggregation. These findings and published data confirm the biological safety of gold nanoparticles for targeted delivery of drugs and phototherapy.

Studies of oxidant-induced changes in albumin transport function with a fluorescent probe K-35. Metal-catalyzed oxidation.

The dynamics of albumin transport function was studied during metal-catalyzed oxidation of albumin in diluted blood plasma from healthy donors and in the solution of purified albumin using fluorescent probe K-35. The changes were compared with the dynamics of free radical oxidation markers. For oxidation, different concentrations of Cu(2+), Fe(2+), Fe(3+) ions as well as EDTA and H(2)O(2) were used. Oxidative modification of proteins was assessed by carbonyl and bityrosine fluorescent products. Oxidation of plasma lipids was assessed by the levels of TBA-reactive products. It was found that oxidation markedly decreased effective concentration of albumin characterizing albumin binding capacity, and leads to accumulation of carbonyl products of protein oxidation, bityrosine fluorescent products in proteins, and TBA-active products of lipid oxidation. It was hypothesized that reduced effective concentration of albumin is related to impairment of its binding sites and/or accumulation of free-radical oxidation products filling the binding sites of albumin.

Studies of oxidant-induced changes in albumin transport function with a fluorescent probe k-35. Effect of hypochlorite.

The dynamics of changes in albumin transport function during hypochlorite-induced oxidation of isolated albumin in blood plasma and serum was studied with a fluorescent probe K-35. Binding of the probe K-35 to albumin was characterized by effective concentration of albumin. Oxidative modification of proteins was evaluated by the content of carbonyl products of protein oxidation and bityrosine fluorescent products. Oxidation with hypochlorite was accompanied by a decrease in the effective concentration of albumin in albumin, diluted plasma, and serum and accumulation of carbonyl products of protein oxidation and bityrosine fluorescent products. The decrease in the effective concentration of albumin during oxidation with hypochlorite can be explained by oxidative damage to albumin binding sites. Oxidative modification of probe K-35 binding sites with hypochlorite contributes to a decrease in effective concentration of albumin under pathological conditions.

Effect of oxidized fibrinogen on aggregation of activated platelets and neutrophils.

The effect of oxidized fibrinogen on platelet-neutrophil complex formation was evaluated by studying the platelet aggregation (changes in light transmission and turbidimetric assay). Activation of cells by thrombin (0.015 U/ml) in the presence of oxidized fibrinogen was accompanied by the formation of larger intermolecular aggregates of platelets and leukocytes as compared to those detected in experiments with non-oxidized fibrinogen. Addition of thrombin (0.2 U/ml) in the presence of oxidized fibrinogen was followed by the formation of more stable complexes of platelets and leukocytes as compared to those revealed in experiments with non-oxidized fibrinogen. An increase in the width of aggregation curves was most pronounced in the system of 10(-4) M Fe(2+) and 10(-4) M H(2)O(2) with oxidized fibrinogen. Our results indicate that oxidized fibrinogen contributes to the "floating" or suspension of platelet-leukocyte complexes.

Oxidative modification of fibrinogen inhibits its transformation into fibrin under the effect of thrombin.

Changes in the capacity of fibrinogen subjected to oxidative modification to transform into fibrin under the effect of thrombin and to form a fibrin clot were studied. The effects of oxidized fibrinogen preparations on the clot formation by citrate-treated donor plasma were evaluated by the thrombin time test. Oxidation impaired the capacity of isolated fibrinogen to form a fibrin clot under the effect of thrombin. Addition of oxidized fibrinogen solutions to donor plasma led to prolongation of the plasma clotting time. Maximum addition (33% volume) of oxidized fibrinogen led to a 10-26% prolongation of clotting time in comparison with addition of the same volume of the same solution without fibrinogen.

Evaluation of hexose monophosphate shunt activity in isolated murine lens by monitoring the potential of the ferricyanide--ferrocyanide system.

A method for evaluation of the activity of the hexose monophosphate shunt (HMS) in isolated lens is presented. The measurement of HMS activity is based on continuous monitoring of the potential of the ferricyanide--ferrocyanide system (where ferricyanide is an artificial electron acceptor) in the presence of a lens. The rate of reduction of ferricyanide increased after the addition of methylene blue (MB) or saponin. The ferricyanide reduction rate did not correlate with GSH content in the contralateral lenses of the same mouse in the absence of MB or saponin. Correlations between the ferricyanide reduction rate and GSH content in the lens were 0.67 (beta = 0.93) in the presence of MB and 0.82 (beta = 0.95) in the presence of saponin. We think that the measured curves of ferricyanide reduction are representative of: 1) normal level of HMS activity (in the absence of methylene blue and saponin); 2) maximal HMS activity (in the presence of methylene blue); 3) the intracellular concentration of reducing equivalents (in the presence of saponin).