Addition of fibrinogen to human umbilical vein endothelial cells in culture resulted in release of von Willebrand factor (vWf) from Weibel-Palade bodies that was temporally related to formation of fibrin in the medium. Whereas no release occurred before gelation, the formation of fibrin was associated with disappearance of Weibel-Palade bodies and development of extracellular patches of immunofluorescence typical of vWf release. Release also occurred within 10 min of exposure to preformed fibrin but did not occur after exposure to washed red cells, clot liquor, or structurally different fibrin prepared with reptilase. Metabolically labeled vWf was immunopurified from the medium after release by fibrin and shown to consist of highly processed protein lacking pro-vWf subunits. The contribution of residual thrombin to release stimulated by fibrin was minimized by preparing fibrin clots with nonstimulatory concentrations of thrombin and by inhibiting residual thrombin with hirudin or heating.We conclude that fibrin formed at sites of vessel injury may function as a physiologic secretagogue for endothelial cells causing rapid release of stored vWf.

We have characterized the effects of ultrasound on fibrinolysis in vitro to investigate the mechanism of ultrasonic potentiation of fibrinolysis and to identify potentially useful ultrasound parameters for therapeutic application. Radiolabeled clots in thin walled tubes were exposed to ultrasound fields in a water bath at 37 degrees C, and lysis was measured by solubilization of radiolabel. Ultrasound accelerated lysis of plasma, whole blood, and purified fibrin clots mediated by recombinant tissue-type plasminogen activator (rt-PA), urokinase, or streptokinase, but ultrasound by itself caused no clot solubilization. The degree of ultrasonic potentiation was dependent on plasminogen activator concentration, increasing from 2.2-fold at a streptokinase concentration of 75 U/mL to 5.5-fold at 250 U/mL in a 1 MHz ultrasound field at 4 W/cm2. Ultrasound exposure resulted in heating due to absorption by the plastic tube, but the temperature increase was insufficient to account for the increase in clot lysis rate, indicating that the primary effect was nonthermal. Ultrasound did not accelerate hydrolysis of a peptide substrate by rt-PA and did not alter the rate of plasmic degradation of fibrinogen, indicating that the augmentation of enzymatic fibrinolysis required the presence of a fibrin gel. The acceleration of fibrinolysis by ultrasound was greater at higher intensities and duty cycles and was maximum at frequencies between 1 and 2.2 MHz, but decreased at 3.4 MHz. These findings suggest that ultrasound accelerates enzymatic fibrinolysis by increasing transport of reactants through a cavitation-related mechanism.

Relatively mobile TcO(+)(-) and NpO(2)(+) can be chemically reduced to less soluble oxidation states in the presence of igneous rocks, as predicted by oxidation-reduction measurements. Current risk assessments, which consider technetium and neptunium as potentially capable of migrating from high-level radioactive waste repositories, may be overestimating their potential hazard to the public since the Fe(II) content of many subsurface waters may maintain these elements in less soluble oxidation states.

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After fibrin polymerization, activated Factor XIII catalyzes the formation of intermolecular cross-links between '-ychain pairs and also among two or more a-chains to form polymers. In this report we characterize the size and heterogeneity of a-chain polymers, establish the role of high concentrations of Factor XIII in determining the extent and rate of a-polymer formation, and also provide evidence that the Factor XIII required can be provided by platelets. Fibrin prepared from purified fibrinogen or platelet-deficient plasma contained a series of cross-linked a-chain polymers with M, from 140,000 to 770,000 with a mean Mr difference of 32,000 consistent with a staggered, overlapping addition of monomers to the growing a-polymer chain. In plasma containing no platelets, a-polymer formation was incomplete with residual a-monomer remaining, but higher platelet counts facilitated more rapid cross-linking into larger polymers. Purified Factor XIII was equally effective as platelets in facilitating cross-linking. We conclude that cross-linked a-polymer chains are heterogeneous in size reaching a molecular weight of several million and that high concentrations of Factor XIII as provided by platelets are required for maximum cross-linking.