Effect of sweet grass extract against oxidative stress in rat liver and serum.

Ethanol metabolism is accompanied by generation of free radicals which can damage the cell components. However, sweet grass is a source of coumarin and its derivatives have emerged as a promising group of antioxidant compounds. The aim of this study has been to investigate the influence of sweet grass on oxidative stress formation in the liver and serum of rats intoxicated with ethanol. Alcohol intoxication led to a decrease in the superoxide dismutase, catalase, glutathione peroxidase and reductase activity in the blood serum as well as in the liver, but not in the glutathione reductase activity. The decrease in the antioxidant abilities of the examined tissues after ethanol intoxication resulted in enhanced lipid peroxidation measured as malondialdehyde and 4-hydroxynonenal levels. The metabolic consequence of oxidative modifications of lipids was damage of the liver cells membrane and an increase in its permeability appeared as a leakage of alanine aminotransferase and aspartate aminotransferase into the blood. Administration of sweet grass to the ethanol-intoxicated rats remarkably prevented the significant increase in concentrations of all measured lipid peroxidation products as well as the damage of the liver cell membrane. These results indicate beneficial antioxidant effect of the sweet grass on the liver of rats intoxicated with ethanol.

Chromatographic determination of fatty acids in biological material.

Fatty acids participate in different metabolic mechanisms, but their key role is connected with their participation in membrane phospholipid organizations. This review introduces the current knowledge of fatty acid analysis with an emphasis on the analysis of these compounds in biological samples. Therefore this manuscript is focused on various aspects of biological sample preparation methods, such as lipid extraction, lipid and phospholipid fractionation (TLC and SPE), derivatization, and on the methodologies of fatty acid analysis by gas chromatography (GC) and high-performance liquid chromatography (HPLC). The wide spectrum of columns and detectors used in these techniques for selective separation and determination of bioactive fatty acids are discussed. The abilities of using a sensitive tandem LC/MS and GC/MS assay are also shown.

The effect of temperature, acidification, and alkalization changes as well as ethanol on salivary cathepsin D activity.

The activity of salivary cathepsin D undergoes inactivation at the temperature of 50-60 degrees C and at pH of 2.0 and pH of 8.0-10.0. The enzyme activity is also decreased by high concentrations of ethanol and high-proof alcoholic beverages. The factors should be taken into consideration in the evaluation of salivary cathepsin D activity.

Effect of chlorhexidine mouthrinse on cathepsin C activity in human saliva.

UNLABELLED: Chlorhexidine is an active agent commonly used against dental plaque in the mouth apart from fluorides applied to prevent caries. It is contained in toothpastes and mouthrinses. PURPOSE: The aim of the study was to assess the effect of mouthrinses containing chlorhexidine digluconate on the activity of cathepsin C in human saliva. MATERIAL AND METHODS: Material for analyses contained mixed saliva samples collected at rest, directly into test tubes (Z PS type, Medlab) at least 2 hours after meal from 40 subjects (dentistry students; 30 women and 10 men), aged 19-24. Saliva was collected before the preparations were applied after rinsing the mouth with distilled water and following a single use of the preparations according to the producer's instructions, 8 samples for each preparation. RESULTS: The decrease of cathepsin C was observed for each preparation, but was the greatest after mouth rinsing with Kin Gingival (65.08%) and Corsodyl (58.00%). CONCLUSIONS: The current study confirms this assumption by finding a decrease in cathepsin C activity after the use of chlorhexidine mouth rinses.

Comparative analysis of the effect of preparations used in professional fluoride prophylaxis on the chosen parameters of human saliva.

UNLABELLED: Regular supply of fluoride ions to the oral environment is one of the prophylactic actions against dental caries. Fluorides, whose exogenous action combines with saliva properties, condition the anticariogenic effect. Fluoride ions exhibit high chemical activity, can alter the oral environment parameters and inhibit the activity of enzymes. PURPOSE: In the current study, the effect of fluoride preparations used in professional caries prophylaxis on chosen saliva parameters was studied. The levels of pH and fluoride ions, and the activity of cathepsin D in human saliva were determined. MATERIAL AND METHODS: Material for analysis contained resting mixed saliva collected before and 1, 4 and 24 hours after the application of Duraphat, Elmex Gel, Fluor Protector, Fluormex Gel and Fluoro-Gel. RESULTS: The fluoride-containing preparations inhibited the activity of cathepsin D in the way depending on the time that had passed since the application and altered the pH level of human saliva.

The choice of conditions for cathepsin D activity determination in human saliva.

The aim of the study was the demonstration and choice of conditions for the determination of cathepsin D activity in human mixed saliva. The 6% solution of hemoglobin, denatured with hydrochloric acid, was used as the substrate. The ratio of saliva volume to hemoglobin was 4:1 w/v. The reaction was interrupted by adding 10% trichloroacetic acid, after 6 hours of incubation at 37 degrees C. The increase in degradation products was determined with the use of Folina and Ciocalteau method with copper modification.

Effect of fluoride preparations on the activity of human salivary cathepsin C.

Preparations containing organic and inorganic fluorine compounds are used for oral hygiene. Fluoride ions contained in these preparations display high bioactivity and can alter the environment of the mouth. The aim of the study was to determine the effect of preparations containing aminofluorides, commonly used in oral hygiene, on the activity of salivary cathepsin C (EC 3.4.14.1). The research material included mixed saliva, collected at rest before and after the application of the following preparations: Elmex gelee, Elmex red fluid, Elmex green fluid, Fluormex rinse. The salivary pH, concentration of fluoride ions and activity of cathepsin C were determined. Fluoride preparations inhibit the activity of cathepsin C and cause changes in human salivary pH. Saliva can serve as a diagnostic material in the examination of the environmental exposure to fluorides.

Immunohistochemical evaluation of mast cells and mark activity tryptase and chymase in experimental fibrosarcoma.

The aim of the study was an evaluation of the activity of mast cells and mark activity tryptase and chymase and of protein levels in experimental fibrosarcoma, induced in rat skin. The experiments were carried out on 50 male Wistar rats. The cancer was induced in rats by one subcutaneous injection of 0.2 mg 3-methylcholanthrene in 0.25 ml of olive oil. Tissue material was fixed in Bouin's fluid. Immunohistochemical tryptase detecting reactions were performed--using specific antibodies and the ABC complex. The activities of tryptase and chymase and protein levels were determined in supernatant of 10% homogenate. We found a very significant growth of mast cell quantity in the connective tissue of tumours. We observed slight differences in the activity of examined enzymes in tumours of different mass.

The activity and location of cathepsin D inhibitor in seeds of common vetch (Vicia sativa L.).

The activity of cathepsin D inhibitor is markedly higher in common vetch seed coat than in embryo cotyledons. The occurrence of considerable amounts of the inhibitor in the seed coat of vetch was confirmed by the fluorescent microscopic technique, with the use of fluorescein-marked cathepsin D.

Effects of cathepsin D inhibitor from Vicia sativa L. seed hulls on human skin fibroblasts and breast cancer cells (in vitro studies).

Cathepsin D is a lysosomal protease which plays an important role in cancer invasion and metastasis. There are known inhibitors of that enzyme, such as pepstatin and potato inhibitor. In this study, we examined effects of the cathepsin D inhibitor from Vicia sativa L. seed hulls on cell cultures of human skin fibroblasts and breast cancer cells. There is no effect of the D-cathepsin inhibitor from Vicia sativa L. seed hulls on the proliferative activity of either human skin fibroblasts or breast cancer cells, measured by the [3H] thymidine incorporation assay.

Synthesis of benzylamides of dipeptides as potential inhibitors of plasmin.

Four benzylamides of dipeptides with the general formula: X-L-Lys-NH-CH2-C6H5, where X = L-or D-Leu and L-or D-Phe were prepared as potential inhibitors of plasmin. All of them influenced on the fibrynolytic activity of plasmin, but only D-Leu-L-Lys-NH-CH2-C6H5 inhibited the amidolytic activity of this enzyme. None of the tested compounds was an inhibitor of thrombin in an amidolytic test.

Effect of epsilon-aminocaproylaminoacids on the activity of proteolytic enzymes.

Effect of three epsilon-aminocaproylaminoacids with significant antifibrinolytic activity on chymotrypsin, trypsin, cathepsin B, cathepsin C and cathepsin D activities was examined. Slight inhibition of trypsin and chymotrypsin activity was observed only at high concentrations of these compounds. All tested dipeptides did not influence activities of cathepsin B, cathepsin C and cathepsin D.

[Activity of cathepsin B and cystatins in the placenta during EPH-gestosis]. / Aktywnosc katepsyny B i aktywnosc cystatyn w lozysku EPH-gestozowym.

Activity of cathepsin B using Bz-DL-Arg-pNA contents of SH-group by means of Ellman method, activity of cystatins against papain using casein as a substrate and contents of deoxyribonuclein acids by Burton method were determined in 64 placentas of pregnancies with EPH-gestosis and in 36 placentas of physiological pregnancies. The placentas from pregnancies with EPH-gestosis showed markedly higher activity of cathepsin B, no difference in the contents of SH-group, slightly higher activity of cystatins and they contain less deoxyribonucleic acids than the placentas from physiological pregnancies. The obtained results show that proteolytic--anti-proteolytic balance in placentas from pregnacies with EPH-gestosis is changed to the advantage of cathepsin B. This protease may in formation of structural and functional changes observed in placentas during EPH-gestosis.

Interaction of L-gamma-glutamyl-alpha,beta-dehydroamino acids with gamma-glutamyl transpeptidase.

Three gamma-glutamyl alpha,beta-dehydroamino acids: L-gamma-glutamyl-dehydroalanine, L-gamma-glutamyl-(Z)-dehydrobutyrine and L-gamma-glutamyl-(E)-dehydrobutyrine have been prepared as potential ligands (inhibitors or substrates) for gamma-glutamyl transpeptidase (GGT). Both isomers of gamma-glutamyl-dehydrobutyrines proved to be inhibitors of GGT, slightly better than the saturated analogue, L-gamma-glutamyl-L-butyrine. However, their solvolysis catalysed by the enzyme is slower than that of the latter. L-gamma-Glutamyl-(E)-dehydrobutyrine seems to be a more active compound in both enzymatic tests. L-gamma-Glutamyl-dehydroalanine elicited only low inhibitory activity and, moreover, was unstable under conditions of the solvolysis test.

Hemorrhagic shock activates mast cells in the rat stomach wall.

The activation of mast cells (MC) and liberation of their mediators can play an essential role in initiating and controlling inflammatory processes in the wall of the gastrointestinal tract (GIT) due to ischemia. The role of MC in changes induced by hemorrhagic shock (HS) remains unknown. Heparin provided by MC seems to inhibit local inflammation and prevent DIC. The aim of this study was to evaluate the morphometric changes and biochemical activity of MC in the stomach wall after 75 minutes of hemorrhagic shock. The MC in mucosal, submucosal, muscular and serosal compartments of the various stomach wall regions were examined in shocked rats and in the control group. Additionally, the contents of glycosaminoglycans (GAG), measured as uronic acids concentration, as well as anticoagulative activity in the stomach wall were assessed. HS resulted in an evident increase in the number of mast cells detected in the stomach mucosa and serosa, in slight alterations in number of MC in the submucosal and muscular layers, a significant increase in size and changes of the shape of the MC. The elevation of the width, area, and circular shape of MC in all layers were noted. No clear and significant differences between various stomach regions in MC numbers and MC sizes could be shown. No reaction of other inflammatory cells at this stage of shock was observed. Highly significant increases in GAG concentration, and anticoagulative activity in the stomach wall due to shock were noted. The morphometric and biochemical data may indicate MC activation, especially in mucosa and serosa. The shock-induced migration of MC settled in the stomach wall seems to be possible. The results suggest an essential role of MC reaction in the stomach wall in the early phase of hemorrhagic shock.

Anticoagulative effect of pepsin.

Anticoagulative effect of pepsin is observed in vitro when its concentration is 36 microM and higher. This effect is due to inhibition of fibrin monomer polymerization. Protamine abolishes anticoagulative effect of pepsin. Pepsin does not influence platelet aggregation induced by ADP and collagen.

The influence of epsilon-aminocaproic acid derivatives on the anticoagulant activity of heparin.

Derivatives of epsilon-aminocaproic acid with antifibrinolytic activity, at low concentration, do not influence the anticoagulant activity of heparin under the heparin-thrombin test conditions. At concentrations higher than 0.002 M tested compounds slightly enhance the anticoagulant action of heparin.

Cathepsin D inhibitor from Vicia sativa L.

Specific inhibitor of cathepsin D has been shown in the extract of Vicia sativa L. seeds. This inhibitor does not inhibit the activity of other aspartic proteases. Also it does not inhibit the activity of cysteine proteases and serine proteases.

Inhibitors of pepsin, trypsin and chymotrypsin in seeds of plants consumed by humans and animals. I. Evaluation of pepsin, trypsin, and chymotrypsin inhibitors activity in seeds of 26 plant species.

Antipepsin, antitrypsin and antichymotrypsin activity was determined in seed extracts of 26 plants consumed by humans and animals (small bean, broad bean, pumpkin, kidney bean, charlock, pea, buckwheat, barley, maize, flax, lupine, poppy, almond, peanut, hazel, walnut, oat, millet, wheat, rice, rape, sunflower, lentils soya bean, vetch, rye). Antipepsin activity is found in the seeds of small bean, pumpkin, flax, peanut, walnut, oat, wheat, sunflower, lentils and soya bean. Antitrypsin and antichymotrypsin activities are of different intensity in seed extracts of all examined plants.