Extraction of plasminogen activator in the rat's submaxillary gland.

Plasminogen is activated into the fibrinolytic enzyme plasmin via: a tissue type activator and F-XII dependent and F-XII independent systems. The purpose of this study was extract and quantify the tissue-type plasminogen activator present in the salivary glands of rats. The extracted plasminogen activator-EPA- was obtained by homogenizing 1 vol of tissue with 1 vol of 2M KSCN solution. Solution with EPA was applied by triplicated in the standard plasminogen-rich and plasminogen-free fibrin plates. The degree of fibrinolytic activity was observed as areas of liquefaction and measured as the product (mm2) of the two perpendicular diameter of the lysed zones. The submaxillary's EPA produced a mean lytic area of 198 mm2 +/- 18 SEM only in the plasminogen-rich fibrin plate. This activity extrapolated into a standard dilution curve, represented the equivalent to a 50 mg/ml plasmin solution. No lysis was induced by EPA from parotid and sublingual glands. The antifibrinolytic drug E-ACA in a dose dependent inhibitory action, significantly reduced the lytic activity induced by submaxillary's EPA. The observation that EPA produced areas of liquefaction only in plasminogen-rich fibrin plate and that this activity was inhibited by E-ACA is clear indication that the zones of lysis was specific fibrinolysis -activation of plasminogen into plasmin- and not due to non-specific proteolysis.

Extraction of plasminogen activator in the rat's submaxillary gland

El objetivo de este estudio fue extraer y cuantificar el activador de plasminógeno AP en glándulas salivares de la rata. AP se extrajo por homogeneización de 1 vol de tejido com 1 vol de una solución de 2M KSCN. La solución con AP fue pipeteada por triplicado en placas de fibrina PF ricas y pobres en plasminógeno. La actividad fibrinolítica inducida por AP se observó como áreas circulares de licuefacción medidas en mm2, resultado del producto de dos diámetros perpendiculares de las zonas de lisis. El AP de la submaxilar produjo un término medio de actividad lítica de 198 mm2 + ou - 18ESM únicamente en PF ricas en plasminógeno. Extrapolando al área de lisis producida por 50 mg/ml de plasmina. No se observó lisis con AP extraído de la parótida y sublingual. La droga antifibrinolítica E-ACA, redujo significantemente, en una acción inhibitoria, dosis dependiente, la actividad fibrinolítica del AP de la submaxilar. El hecho de que AP indujo lisis únicamente en PF ricas en plasminógeno, la que fue significativamente reducida por E-ACA, indica claramente que la actividad lítica observada en el sustrato fibrina es debido a una fibrinolisis específica y no debido a una proteólisis no específica

Extraction of plasminogen activator in the rats submaxillary gland.

Plasminogen is activated into the fibrinolytic enzyme plasmin via: a tissue type activator and F-XII dependent and F-XII independent systems. The purpose of this study was extract and quantify the tissue-type plasminogen activator present in the salivary glands of rats. The extracted plasminogen activator-EPA- was obtained by homogenizing 1 vol of tissue with 1 vol of 2M KSCN solution. Solution with EPA was applied by triplicated in the standard plasminogen-rich and plasminogen-free fibrin plates. The degree of fibrinolytic activity was observed as areas of liquefaction and measured as the product (mm2) of the two perpendicular diameter of the lysed zones. The submaxillarys EPA produced a mean lytic area of 198 mm2 +/- 18 SEM only in the plasminogen-rich fibrin plate. This activity extrapolated into a standard dilution curve, represented the equivalent to a 50 mg/ml plasmin solution. No lysis was induced by EPA from parotid and sublingual glands. The antifibrinolytic drug E-ACA in a dose dependent inhibitory action, significantly reduced the lytic activity induced by submaxillarys EPA. The observation that EPA produced areas of liquefaction only in plasminogen-rich fibrin plate and that this activity was inhibited by E-ACA is clear indication that the zones of lysis was specific fibrinolysis -activation of plasminogen into plasmin- and not due to non-specific proteolysis.

Extraction of plasminogen activator in the rats submaxillary gland

El objetivo de este estudio fue extraer y cuantificar el activador de plasminógeno AP en glándulas salivares de la rata. AP se extrajo por homogeneización de 1 vol de tejido com 1 vol de una solución de 2M KSCN. La solución con AP fue pipeteada por triplicado en placas de fibrina PF ricas y pobres en plasminógeno. La actividad fibrinolítica inducida por AP se observó como áreas circulares de licuefacción medidas en mm2, resultado del producto de dos diámetros perpendiculares de las zonas de lisis. El AP de la submaxilar produjo un término medio de actividad lítica de 198 mm2 + ou - 18ESM únicamente en PF ricas en plasminógeno. Extrapolando al área de lisis producida por 50 mg/ml de plasmina. No se observó lisis con AP extraído de la parótida y sublingual. La droga antifibrinolítica E-ACA, redujo significantemente, en una acción inhibitoria, dosis dependiente, la actividad fibrinolítica del AP de la submaxilar. El hecho de que AP indujo lisis únicamente en PF ricas en plasminógeno, la que fue significativamente reducida por E-ACA, indica claramente que la actividad lítica observada en el sustrato fibrina es debido a una fibrinolisis específica y no debido a una proteólisis no específica (AU)

Extraction and quantification of heparin in "clinically normal" and inflamed gingiva of the dog.

Heparin present in tissue has been reported to be a potential locally active agent responsible for bone resorption by the stimulation of collagenase via osteoclast activation and increased collagenase synthesis by the osteoblast. The purpose of this study was to extract, identify and quantify heparin in the "clinically normal", mildly and severely inflamed gingiva of Beagle dogs, using a simple and reliable technique. The extraction was carried out by homogenization, fat elimination, proteolytic digestion and precipitation, with organic solvents. Identification was performed by microelectrophoresis conducted on an agarose coated microscope slide. Quantification was performed by measuring the optical density of the metachromatic toluidine blue stained spot an comparing with the standard reference curve of heparin run simultaneously. The results showed that the difference in concentration of heparin in units per gram of wet tissue, was not significant when the "clinically normal" (26 +/- 1.9) was compared with mildly inflamed (24.4 +/- 4.7) gingiva. However, the concentration of heparin in severe gingivitis (79 +/- 7) was significantly higher. Gingival heparin could play important role in the established periodontal lesions, acting as a local factor or co-factor in periodontal bone destruction.

Extraction and quantification of heparin in [quot ]clinically normal[quot ] and inflamed gingiva of the dog.

Heparin present in tissue has been reported to be a potential locally active agent responsible for bone resorption by the stimulation of collagenase via osteoclast activation and increased collagenase synthesis by the osteoblast. The purpose of this study was to extract, identify and quantify heparin in the [quot ]clinically normal[quot ], mildly and severely inflamed gingiva of Beagle dogs, using a simple and reliable technique. The extraction was carried out by homogenization, fat elimination, proteolytic digestion and precipitation, with organic solvents. Identification was performed by microelectrophoresis conducted on an agarose coated microscope slide. Quantification was performed by measuring the optical density of the metachromatic toluidine blue stained spot an comparing with the standard reference curve of heparin run simultaneously. The results showed that the difference in concentration of heparin in units per gram of wet tissue, was not significant when the [quot ]clinically normal[quot ] (26 +/- 1.9) was compared with mildly inflamed (24.4 +/- 4.7) gingiva. However, the concentration of heparin in severe gingivitis (79 +/- 7) was significantly higher. Gingival heparin could play important role in the established periodontal lesions, acting as a local factor or co-factor in periodontal bone destruction.

Extraction and quantification of heparin in [quot ]clinically normal[quot ] and inflamed gingiva of the dog

Heparin present in tissue has been reported to be a potential locally active agent responsible for bone resorption by the stimulation of collagenase via osteoclast activation and increased collagenase synthesis by the osteoblast. The purpose of this study was to extract, identify and quantify heparin in the [quot ]clinically normal[quot ], mildly and severely inflamed gingiva of Beagle dogs, using a simple and reliable technique. The extraction was carried out by homogenization, fat elimination, proteolytic digestion and precipitation, with organic solvents. Identification was performed by microelectrophoresis conducted on an agarose coated microscope slide. Quantification was performed by measuring the optical density of the metachromatic toluidine blue stained spot an comparing with the standard reference curve of heparin run simultaneously. The results showed that the difference in concentration of heparin in units per gram of wet tissue, was not significant when the [quot ]clinically normal[quot ] (26 +/- 1.9) was compared with mildly inflamed (24.4 +/- 4.7) gingiva. However, the concentration of heparin in severe gingivitis (79 +/- 7) was significantly higher. Gingival heparin could play important role in the established periodontal lesions, acting as a local factor or co-factor in periodontal bone destruction.

Determination of heparin in clinically normal dog gingiva.

The purpose of this study was to develop a technique to allow the extraction, identification and quantification of heparin in small samples (0.5 g) of normal gingival tissue of the Beagle dog. Heparin was extracted following homogenization, defattening, proteolytic digestion and precipitation with organic solvents and identified by a microelectrophoretic method carried out on an agarose slide. The concentration of heparin was determined by measuring the optical density of the metachromatic toluidine blue stained spots and comparing them with standard reference heparin spots. Gingiva contained 25 +/- 1.9 units of heparin per gram of wet tissue with a relative migration rate of 0.92 +/- 0.005 vs the unit value for reference heparin. Heparin was further characterized by enzyme degradation with adapted F. heparinum (heparinase). Its anticoagulant activity was demonstrated by extending Plasma Thrombin Time, which was normalized with the anti-heparin substance protamine sulfate.