CLEANSOIL as a perspective method of remediation of oil-contaminated soils under existing infrastructure

The results are given of the two-year study of the new technology of soil remediation from oil contamination-CLEANSOIL combining physicochemical and biological methods of eliminating oil pollution. It is shown that using this combined technology results in quick decrease in the oil concentration in soil to the level when the soil is able to clean itself.     

Hypochlorous acid-induced lysis of human erythrocytes. Inhibition of cellular damage by the isoflavonoid genistein-8-C-glucoside

Erythrocyte damage induced by hypochlorous acid (HOCl) results in cell lysis developing with time after the oxidant is removed (post-hemolysis). The apparent rate constant of post-hemolysis depends on time of incubation in the presence of HOCl and concentration of this oxidant. HOCl-dependent damage of erythrocyte membranes is associated with uncompetitive inhibition of the membrane-bound acetylcholinesterase. Genistein-8-C-glucoside is an isoflavonoid isolated from the flowers of Lupinus luteus L.; in aqueous solution, genistein-8-C-glucoside (0.5-2 mM) efficiently inhibited HOCl-induced damage to erythrocytes similar to the known HOCl scavengers taurine and reduced glutathione. This bioflavonoid can protect the erythrocyte membrane (and to a lesser extent, intraerythrocytic components) by interacting with the reactive chlorine species including hypochlorous acid and membrane-bound chloroamines formed in the reaction of HOCl with erythrocyte membrane proteins.     

Effect of physical exercise on fibrinolytic properties of erythrocytes

The fibrinolytic properties of blood and erythrocytes were studied before and after physical exercise in male volunteers. Their fibrinolytic responses were of two distinct types. In type 1 response, fibrinolytic activities of blood and erythrocytes increased; the plasminogen activator and active plasmin contents in erythrocytes also increased, whereas the profibrinolysin content correspondingly decreased. In addition, physical exercise increased the erythrocyte adsorption properties for plasma activators of fibrinolysis. Type 2 response was characterized by a decrease in the fibrinolytic activity of blood; neither fibrinolytic activity nor adsorption properties of erythrocytes increased. The type of blood and erythrocyte response to muscular activity was determined by the pre-exercise level of red blood cell fibrinolytic activity. It was low in type 1 response due to a lesser content of plasmin activators and greater content of antiplasmin. In type 2 response, the initially high lytic capacity is connected with a greater reserve of activators and lesser reserve of inhibitors of the fibrinolytic system. A conclusion was made that individual differences in fibrinolytic responses to physical exercise were largely accounted for by the properties of erythrocytes.     

Method for determining kallikrein of tissue origin in blood plasma and its clinical significance

Based on a study of the kininogenase activity of the total plasma kallikrein in the presence of 3 concentrations of the soybean inhibitor trypsin (0.5, 1.0, 10.0 micrograms/ml) one can measure at a time the activity of tissue kallikrein (without specifying the source) and the activity of 3 forms of plasma kallikrein, including its adsorption on kaolin that characterizes the conformational structure of the enzyme. Examination of 10 healthy subjects and 136 patients revealed a 10 to 20-fold increase in the content of tissue kallikrein in plasma of 70% of diabetes mellitus patients and a 2.5 to 3-fold elevation in 50% of patients with chronic occupational bronchitis, and in 30% of patients suffering from chronic hepatitis. The method suggested makes it possible to have a better insight into the physiological and pathogenetic role of the kinin system and may be used for laboratory control over the treatment efficacy.     

Corrections by melatonin of liver mitochondrial disorders under diabetes and acute intoxication in rats

The aim of the present work was to investigate the mechanisms of oxidative damage of the liver mitochondria under diabetes and intoxication in rats as well as to evaluate the possibility of corrections of mitochondrial disorders by pharmacological doses of melatonin. The experimental (30 days) streptozotocin‐induced diabetes mellitus caused a significant damage of the respiratory activity in rat liver mitochondria. In the case of succinate as a respiratory substrate, the ADP‐stimulated respiration rate V₃ considerably decreased (by 25%, p < 0·05) as well as the acceptor control ratio (ACR) V₃/V₂ markedly diminished (by 25%, p < 0·01). We observed a decrease of the ADP‐stimulated respiration rate V₃ by 35% (p < 0·05), with glutamate as substrate. In this case, ACR also decreased (by 20%, p < 0·05). Surprisingly, the phosphorylation coefficient ADP/O did not change under diabetic liver damage. Acute rat carbon tetrachloride‐induced intoxication resulted in considerable decrease of the phosphorylation coefficient because of uncoupling of the oxidation and phosphorylation processes in the liver mitochondria. The melatonin administration during diabetes (10 mg·kg^‐1 body weight, 30 days, daily) showed a considerable protective effect on the liver mitochondrial function, reversing the decreased respiration rate V₃ and the diminished ACR to the control values both for succinate‐dependent respiration and for glutamate‐dependent respiration. The melatonin administration to intoxicated animals (10 mg·kg⁻¹ body weight, three times) partially increased the rate of succinate‐dependent respiration coupled with phosphorylation. The impairment of mitochondrial respiratory plays a key role in the development of liver injury under diabetes and intoxication. Melatonin might be considered as an effector that regulates the mitochondrial function under diabetes. Copyright © 2011 John Wiley & Sons, Ltd.     

Electron microscopic study of the chitosan effect on the intracellular accumulation and the state of tobacco mosaic virus particles in tobacco leaves

Influence of chitosan on the accumulation and state of tobacco mosaic virus (TMV) in the mesophyll cells of Nicotiana tabacum L. var Samsun leaves in early period of infection development (3 days after infection of leaves) has been studied. The virus accumulated in the cells of the leaves treated for 24 h before infection with chitosan to a lesser degree than in the control cells. The chitosan affected the formation of TMV-specific granular and tubular inclusions which are known to consist of the viral replicase components. Three days after infection of the leaves treated with the chitosan, a typical sign of the infection development was the predominant formation of granular inclusions which are known to appear at the early stages of TMV replication. The infected cells of the leaves untreated with chitosan contained mainly tubular inclusions which had been shown previously to be formed from granular ones at the last stages of the infection process. This indicates that chitosan treatment of the leaves leads to a delay of the development of infection. In phosphotungstic acid-stained suspensions obtained from the infected leaves, abnormal (swollen and "thin") TMV particles were observed along with normal ones. The appearance of abnormal virus particles seems to be caused by virus-induced activation of intracellular lytic processes. The most lytic activity in the infected cells as well as the highest number of abnormal viral particles was observed under the chitosan action. Therefore, it appears that chitosan-mediated stimulation of lytic processes causing destruction of TMV particles may be one of the protective mechanisms limiting virus accumulation in cells.     

Effect of fucoidan from brown alga <Emphasis Type="Italic">Fucus evanescens</Emphasis> on a formation of TMV-specific inclusions in the cells of tobacco leaves

The effect of fucoidan (1.3; 1.4-α-L-fucan), a sulfated polysaccharide from the brown alga Fucus evanescens on the formation of specific granular and tubular inclusions induced by tobacco mosaic virus (TMV) and consisted presumably of the virus-coded protein components of the viral replicase was investigated in the TMV-infected leaves of tobacco (Nicotiana tabacum L.). In four days after inoculation of the leaves with a TMV preparation (1 mg/ml), the signature of infection in a presence of fucoidan (1 mg/ml) was a preferential formation of intracellular granular inclusions, which were related to early stages of the virus reproduction. When infected leaves were not treated with fucoidan, their cells contained mainly tubular inclusions, which were presumably formed from the granular ones on the last stages of the infection process. These observations demonstrated that fucoidan delayed the development of the TMV-induced infection.     

Effect of chitosan on tobacco mosaic virus(TMV) accumulation,hydrolase activity,and morphological abnormalities of the viral particles in leaves of N. tabacum L. cv. Samsun

The effect of chitosan on the development of infection caused by Tobacco mosaic virus(TMV) in leaves of Nicotiana tabacum L. cv. Samsun has been studied. It was shown that the infectivity and viral coat protein content in leaves inoculated with a mixture of TMV(2 μg/mL) and chitosan(1 mg/mL) were lower in the early period of infection(3 days after inoculation), by 63% and 66% respectively, than in leaves inoculated with TMV only. Treatment of leaves with chitosan 24 h before inoculation with TMV also caused the antiviral effects, but these were less apparent than when the virus and polysaccharide were applied simultaneously. The inhibitory effects of the agent decreased as the infection progressed. Inoculation of leaves with TMV together with chitosan considerably enhanced the activity of hydrolases(proteases, RNases) in the leaves, in comparison with leaves inoculated with TMV alone. Electron microscope assays of phosphotungstic acid(PTA)-stained suspensions from infected tobacco leaves showed that, in addition to the normal TMV particles(18 nm in diameter, 300 nm long), these suspensions contained abnormal(swollen, "thin" and "short") virions. The highest number of abnormal virions was found in suspensions from leaves inoculated with a mixture of TMV and chitosan. Immuno-electron microscopy showed that "thin" virus particles, in contrast to the particles of normal diameter, lost the ability to bind to specific antiserum. It seems that the chitosan-induced activation of hydrolases stimulates the intracellular degradation of TMV particles and hence hydrolase activation may be considered to be one of the polysaccharide-mediated cellular defense mechanisms that limit virus accumulation in cells.     

Protective effects of N-acetyl-L-cysteine against acute carbon tetrachloride hepatotoxicity in rats

In recent years, N-acetyl-L-cysteine (NAC) has been widely investigated as a potentially useful protective and antioxidative agent to be applied in many pathological states. The aim of the present work was further evaluation of the mechanisms of the NAC protective effect under carbon tetrachloride-induced acute liver injuries in rats. The rat treatment with CCl4 (4 g/kg, intragastrically) caused pronounced hepatolysis observed as an increase in blood plasma bilirubin levels and hepatic enzyme activities, which agreed with numerous previous observations. The rat intoxication was accompanied by an enhancement of membrane lipid peroxidation (1.4-fold) and protein oxidative damage (protein carbonyl group and mixed protein-glutathione disulphide formations) in the rat liver. The levels of nitric oxide in blood plasma and liver tissue significantly increased (5.3- and 1.5-fold, respectively) as blood plasma triacylglycerols decreased (1.6-fold). The NAC administration to control and intoxicated animals (three times at doses of 150 mg/kg) elevated low-molecular-weight thiols in the liver. The NAC administration under CCl4-induced intoxication prevented oxidative damage of liver cells, decreased membrane lipid peroxidation, protein carbonyls and mixed protein-glutathione disulphides formation, and partially normalized plasma triacylglycerols. At the same time the NAC treatment of intoxicated animals did not produce a marked decrease of the elevated levels of blood plasma ALT and AST activities and bilirubin. The in vitro exposure of human red blood cells to NAC increased the cellular low-molecular-weight thiol levels and retarded tert-butylhydroperoxide-induced cellular thiol depletion and membrane lipid peroxidation as well as effectively inhibited hypochlorous acid-induced erythrocyte lysis. Thus, NAC can replenish non-protein cellular thiols and protect membrane lipids and proteins due to its direct radical-scavenging properties, but it did not attenuate hepatotoxicity in the acute rat CCl4-intoxication model.