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Biomedical subjects

F Bachmann

Publications and source records attributed to F Bachmann.

At least 91 records · Page 5Linked to original sources
PubMed1986

Studies on the release of a plasminogen activator inhibitor by human platelets.

The relative contribution of platelets to plasminogen activator inhibitor (PA-inhibitor) activity in blood was investigated. From the difference in PA-inhibitor levels in platelet-poor plasmas of 12 donors (3 +/- 1 U/ml, mean +/- 95% confidence limits) and in the corresponding platelet-rich plasmas after induction of platelet aggregation by collagen, ADP or epinephrine (7 +/- 1 U/ml), it may be concluded that a greater amount of PA-inhibitor in blood is associated with platelets than with plasma. In collagen-stimulated platelets maximal release of PA-inhibitor and of beta-thromboglobulin (beta-TG) was attained within fifteen seconds, whereas in ADP-stimulated platelets the release of both factors was slower. In platelet-poor plasma no correlation was found between the level of PA-inhibitor and that of beta-TG. Thus, the PA-inhibitor found in plasma is not derived from platelets that had been stimulated after blood collection. The rate of complex formation and the Mr of the principal complexes of radioiodinated tissue-type plasminogen activator (t-PA) or urokinase (UK), in platelet-poor plasma, in platelet-rich plasma after platelet aggregation or in an extract of washed platelets was the same. Moreover, complexes of UK or t-PA with plasmatic PA-inhibitor or with the PA-inhibitor(s) from platelets bound to immobilized antibodies against bovine endothelial cell-derived PA-inhibitor. These results show that the PA-inhibitors in plasma and in platelets are very similar or identical.

Adenosine Diphosphate↗
PubMed1986

The fast-acting inhibitor of tissue-type plasminogen activator in plasma is also the primary plasma inhibitor of urokinase.

We have compared the ability of a plasminogen activator inhibitor (PA-inhibitor) in human plasma, to form complexes with radioiodinated tissue-type plasminogen activator (t-PA) and high molecular weight urokinase (HMr-UK). Addition of 125I-t-PA (final concentration 10 IU/ml) or of 125I-HMr-UK (2 IU/ml) to a plasma containing 33 U/ml of PA-inhibitor resulted in the rapid formation of a 110,000 Mr complex of 125I-t-PA or a 95,000 Mr complex of 125I-HMr-UK with PA-inhibitor. Upon prolonged incubation of the plasma with 125I-HMr-UK a secondary complex of a Mr of 88,000 was observed, which probably derives from limited degradation of the 95,000 complex. Preincubation of the plasma with unlabelled t-PA, HMr-UK or LMr-UK at higher concentrations prevented the subsequent formation of complexes between radiolabelled PAs and the PA-inhibitor. These results thus demonstrate that t-PA and UK form complexes with the same PA-inhibitor. The rate of complex formation of 125I-t-PA or of 125I-HMr-UK with the plasma PA-inhibitor was similar (second order rate constant of association with PA-inhibitor in the order of 10(7) M-1s-1).

Glycoproteins↗
PubMed1986

Human primary colon carcinomas xenografted into nude mice. I. Characterization of plasminogen activators expressed by primary tumors and their xenografts.

Analysis was made of plasminogen activator (PA) activities present in 0.125% Triton X-100 extracts of human primary colon carcinomas and of their respective serial subcutaneous xenografts in nude mice. A correlation between tumor invasiveness and PA expression was observed in that primary tumors exhibiting clearly invasive growth patterns demonstrated high concentrations of PAs while subcutaneous xenografts, exhibiting noninvasive pseudobenign growth, contained very low levels of PA activity. The decrease in fibrinolytic activity observed in subcutaneous xenografts was not due to an increase in inhibitors of fibrinolytic activity. Immunologic characterization of PAs in tumor extracts showed that over 90% of human PA activity was of the urokinase type. Furthermore, tumor-derived urokinase was shown to be present in a proenzyme form. It was resistant to diisopropyl fluorophosphate (DFP) and was not inhibited by purified PA inhibitor. However, after its activation into urokinase by plasmin, it was completely inhibited by DFP and PA inhibitor.

Adenocarcinoma↗
PubMed1986

Human primary colon carcinomas xenografted into nude mice. II. Modulation of tumor plasminogen activator activity by the host tissue environment.

The characterization and quantitation of plasminogen activators (PAs) expressed by human colon carcinoma cell lines and primary colon carcinomas inoculated into nude outbred (nu/nu) mice, either as subcutaneous or gut-implanted (GI) xenografts, were discussed. The two colon carcinoma cell lines used in this study, Col 112 (moderately differentiated) and Col 115 (poorly differentiated), differ in their PA expression, the former being a urinary-type PA and the latter being a tissue-type PA producer. Both cell lines demonstrate a positive correlation between tumor invasiveness and measured PA activity; subcutaneous xenografts growing as noninvasive pseudobenign tumor masses were associated with low levels of PA activity, whereas GI xenografts exhibiting invasive growth expressed higher PA activity. Furthermore, coinoculation of Col 115 tumor cells sc and GI in the same host induced high levels of PA activity in subcutaneous xenografts, suggesting a stimulatory effect of the GI xenograft on subcutaneous xenograft PA expression. Also, purified murine plasminogen was demonstrated to represent an efficient substrate for tumor-secreted human PA.

Animals↗
PubMed1985

[Willebrand's disease and multimeric analysis of the Willebrand factor].

The multimeric composition of von Willebrand factor (vWF) in the plasma of 20 von Willebrand's disease patients was analyzed to determine the type of von Willebrand's disease and to evaluate retrospectively the predictive value of the classical tests. Analysis of the multimeric pattern revealed that 3 had type II and the other 17 type I von Willebrand's disease. The classical tests, especially crossed immunoelectrophoresis of vWF and the ristocetin-induced platelet agglutination test, did not permit correct classification but led to overestimation of the type II von Willebrand variant. Multimeric analysis of von Willebrand factor is necessary to diagnose von Willebrand's disease type II, in which DDAVP infusions should not be given since they are ineffective or may cause thrombocytopenia.

Blood Protein Electrophoresis↗
PubMed1985

Human tissue-type plasminogen activator. Production in continuous serum-free cell culture and rapid purification.

A simplified procedure for the production and purification of human tissue-type plasminogen activator (t-PA) is described. Bowes-melanoma cells were maintained in continuous serum-free culture. The cell nutrient consisted of Dulbecco's modified Eagle's medium (DMEM) supplemented with insulin (5 mg/litre), transferrin (5 mg/litre), progesterone (1 nM), cortisol (10 nM), aprotinin (2 X 10(4) units/litre) and a mixture of trace elements. t-PA accumulated in the culture medium at a rate of 40 units/day per ml and was harvested every third day. Cell losses during each harvest, leading to a steady decline of enzyme yields, were compensated for by treating the cells with 5% (v/v) fetal-bovine serum in DMEM every 6-8 weeks. t-PA was rapidly purified by a combination of cation-exchange chromatography and gel filtration. The procedure yielded mainly single-chain t-PA of a specific activity of 80 000 to 100 000 units/mg.

Blood↗
PubMed1985

Profound alterations of the multimeric structure of von Willebrand factor in a patient with malignant lymphoma.

Laboratory investigation of an acquired haemorrhagic diathesis in a 63-year-old man with malignant lymphoma revealed the classical haemostatic defects found in von Willebrand's disease (vWD). In addition, SDS-agarose gel electrophoresis demonstrated alterations of the von Willebrand factor (vWF) multimeric structure. A profound defect of large and intermediate size multimers was observed which was different from those seen in variants of congenital vWD. In vitro, weak inhibitory activity against factor VIII procoagulant activity and ristocetin cofactor activity was present in the patient's plasma. When patient's plasma was incubated with normal plasma, followed by centrifugation, vWF antigen (vWF:Ag) was precipitated. In vivo, after transfusion of cryoprecipitate, there was rapid plasma clearance of vWF:Ag and ristocetin cofactor and of FVIII coagulant activities.

Factor VIII↗
PubMed1985

Release of vascular plasminogen activator (v-PA) after venous stasis: electrophoretic-zymographic analysis of free and complexed v-PA.

To better characterize the plasminogen activators present in the euglobulin fraction (EF) of plasma before and after venous occlusion, euglobulins of 30 healthy volunteers and of 32 patients with idiopathic thromboembolic disease were submitted to SDS-PAGE and zymographic detection. The patterns thus obtained were compared to the fibrinolytic activity (FA) of the EFs measured on fibrin plates. There were significantly more patients with a low fibrinolytic response (FR) to stasis (difference between FA after and before stasis less than or equal to 0.3 TAU/ml) than controls (P less than 0.01). The electrophoretic-zymographic analysis revealed that: (i) in the resting state vascular plasminogen activator (v-PA) is always present in a form exhibiting a mol wt of 110 000 (complex of v-PA with its fast-acting antiactivator); (ii) a high FR to stasis (greater than 1.3 TAU/ml) is always associated with the presence of free v-PA (68 kdalton band); (iii) free v-PA is never detected when the FR is low; and (iv) a poor FR is generally (15 of 17 patients with low FR) associated with apparently complete inhibition of the released v-PA by the fast-acting antiactivator; in the two remaining patients in whom no broadening of the 110 kdalton band is observed, the release mechanism for v-PA is probably impaired.

Adolescent↗
PubMed1984

Characterization of plasminogen activators from normal human breast and colon and from breast and colon carcinomas.

Triton X-100 and NaSCN extracts of 18 normal breast and colon tissues and of 20 breast and colon carcinomas were fractionated by SDS-PAGE and plasminogen activators (PA) revealed by a zymographic method. Four different lysis bands, corresponding to MWs of 54,000, 68,000, 95,000 and 110,000 were observed. Using immunoadsorption with specific antisera against urokinase (UK) and tissue PA (t-PA), we found that all normal tissue extracts contained free t-PA (68 kd). Some of these revealed, in addition, a complex (110 kd) of t-PA with a 40-kd component. The latter presumably represents the fast-acting specific inhibitor of t-PA and UK. Most carcinoma extracts contained, in addition to the two t-PA-related lysis bands, the UK-related 54 kd PA, and some a 95 kd complex of UK with the 40 kd component. For each extractant, mean total fibrinolytic activity of normal and tumor tissue was comparable when measured on conventional fibrin plates, but breast and colon carcinomas contained higher concentrations of UK-related PA. PA activity was higher in normal and carcinoma NaSCN extracts than in the corresponding Triton X-100 extracts. In general, Triton X-100 but not NaSCN extracts of malignant tissue contained a high concentration of fibrinolytic inhibitors. Mixing experiments revealed that the inhibitory activity was mainly directed against UK. It was abolished by acidification of the carcinoma extracts. The anti-UK inhibitory activity was absent in extracts of normal breast or colon and appears to be different from the 40 kd fast-acting PA inhibitor. These studies show that malignant transformation of breast and colon is accompanied by important changes of the production of a UK-related PA and of an inhibitory activity directed against UK.

Breast↗
PubMed1984

Fibrinolytic activity in healthy volunteers before and after 5 to 20 minutes of venous occlusion.

We have determined in 22 volunteers the fibrinolytic activities of regular and of dextran sulfate precipitated euglobulin fractions in the presence and absence of flufenamate before and after 5 to 20 min of venous occlusion. Using the fibrin plate assay, Hageman factor Independent Intrinsic Fibrinolytic activity ( HIIF ) was measured in plasma euglobulins obtained from resting subjects in the morning. Hageman factor Dependent Intrinsic Fibrinolysis ( HDIF ) was calculated by subtracting the HIIF from the activity obtained after activation of the "resting plasma" by dextran sulfate. The activity of vascular Plasminogen Activator (v-PA) released after venous stasis was determined by subtraction of resting HIIF from post-occlusion total plasminogen activator activity. In normal subjects, the distribution of all activities followed a logarithmic scale. Twelve of 22 volunteers exhibited maximal v-PA activity after 20 min of venous occlusion. In three, it was maximal after 5 min, in three others after 10 min, and in four after 15 min of venous stasis. These results demonstrates that it may not be possible to clearly distinguish normal from abnormal responders using a stasis test of fixed duration.

Adolescent↗
PubMed1984

Effects of hypoxia and acidosis on vascular plasminogen activator release in the pig ear perfusion system.

The influence of hypoxia and acidosis on the release of vascular plasminogen activator was studied in the isolated perfused pig ear. Perfusion with deoxygenated or acid solutions led to significant increases in plasminogen activator release (p less than 0.001); acid solutions of pH 6.7 were more effective than those of pH 6.9. Perfusions with lactate and AMP, two metabolic products which might act as mediators of fibrinolytic stimulation during hypoxia, resulted in moderate increases of plasminogen activator in the effluent (p less than 0.01).

Acid-Base Equilibrium↗
PubMed1984

Tissue-type plasminogen activator increases the binding of glu-plasminogen to clots.

Porcine tissue-type plasminogen activator (t-PA) increases the binding of 125I-glu-plasminogen to clots made from human plasma or purified fibrinogen in a time and t-PA concentration dependent fashion. The accumulation of plasminogen was faster and greater on noncrosslinked plasma clots than on clots which had been crosslinked by Factor XIIIa. Furthermore, the uptake of plasminogen to crosslinked fibrin clots occurred at a slower rate in the presence of alpha 2-plasmin inhibitor (alpha 2 PI) than in its absence. The kinetics of the uptake of 125I-plasminogen were analyzed using SDS-polyacrylamide gel electrophoresis and radioautography of solubilized plasma clots formed in the presence of t-PA. During the initial phase there was a decrease of clot-bound glu-plasminogen; simultaneously, there was a slight increase in clot-bound glu-plasmin and in plasmin complexed to alpha 2 PI that was crosslinked to alpha-chain polymers of fibrin. This was followed by a marked increase in clot-bound plasminogen having glutamic acid as NH2-terminal (glu-plasminogen) and gluplasmin. t-PA-induced enhancement of glu-plasminogen uptake appears to be mediated by plasmin but does not require the conversion of glu-plasminogen to plasminogen having lysine or methionine as NH2-terminal. The described mechanism assures an adequate supply of clot-bound plasmin, which is the enzyme ultimately involved in the degradation of fibrin.

Animals↗
PubMed1984

Pharmacokinetic study of gentamicin-loaded cement in total hip replacements. Comparative effects of varying dosage.

A randomised, double-blind study was performed in two groups of 15 patients undergoing total hip replacements, using antibiotic-loaded acrylic cement containing 0.5 g and 1.0 g gentamicin base respectively per 40 g pack of powdered polymer. Postoperatively, the gentamicin levels in the blood, in the urine and in the wound drainage fluid were measured. In both groups of patients, the serum gentamicin concentrations were low whereas the wound drainage fluid contained highly effective antibacterial concentrations. Serum, urine and wound secretion levels showed approximately two-fold higher concentrations in the group of patients receiving the higher gentamicin load.

Bone Cements↗
PubMed1984

Demonstration of a fast-acting inhibitor of plasminogen activators in human plasma.

This report describes a plasmatic, fast-acting, specific inhibitor (antiactivator) of tissue-type plasminogen activator (t-PA) and urokinase (UK). After addition of t-PA to human plasma, biexponential decay of activity occurred. The initial rapid inhibition of t-PA activity (half-life of approximately one minute) was correlated with the formation of a complex of a molecular weight of 110,000, suggesting a molecular weight in the order of 40,000 for the antiactivator. Diisopropylfluorophosphate (DFP)-inactivated t-PA did not form complexes with antiactivator. The second-order rate constant for the interaction of t-PA with antiactivator is in the order of 10(7) mol/L-1 sec-1. In plasma, UK added at low concentrations rapidly formed complexes of a mol wt of 95,000. Preincubation of the plasma with t-PA prevented complex formation of UK, and vice versa, suggesting that the same inhibitor inactivates both t-PA and UK. After exhaustion of the antiactivator, t-PA and UK formed complexes with alpha 2-antiplasmin and C1'-inhibitor at a low rate.

Adult↗
PubMed1984

The mechanism of in vitro clot lysis induced by vascular plasminogen activator.

The contribution of vascular plasminogen activator (v-PA) to the lysis of whole blood and plasma clots was investigated. v-PA released into the circulation after infusion of deamino-D-arginine vasopressin (DDAVP) was shown to bind quantitatively to plasma clots. Its apparent molecular weight, determined by the SDS-PAGE fibrin-agarose underlay method, was approximately 68,000 daltons, and its activity was quenched by antibodies against human tissue plasminogen activator (t-PA). Clots prepared from post-DDAVP plasma or post-DDAVP whole blood, rich in v-PA, did not lyse when incubated in imidazole buffer or normal plasma, as determined by the release of 125I from radiolabeled clots. However, clots made of v-PA-poor plasma or whole blood, incubated in v-PA-rich plasma, underwent substantial lysis. The concentration of PA in clots incubated in v-PA-rich plasma progressively increased in relation to the initial concentration of v-PA in the surrounding plasma. The results suggest that, at low concentrations of circulating v-PA, a hemostatic plug will lyse at a very low rate. However, when the v-PA concentration in the clot environment is increased, v-PA will accumulate progressively onto fibrin and induce thrombolysis.

Calcium↗