scholarly journals Mini-plasminogen: a mechanism for leukocyte modulation of plasminogen activation by urokinase

Blood ◽  
1981 ◽  
Vol 58 (1) ◽  
pp. 97-104
Author(s):  
LA Moroz
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Gel Filtration ◽  
Aminocaproic Acid ◽  
Binding Activity ◽  
Major Product ◽  
Plasminogen Activation ◽  
Leukocyte Elastase ◽  
Binding Function ◽  
Plasmin Inhibitors ◽  
Epsilon Aminocaproic Acid

Urokinase activation of blood fibrinolysis involves polymorphonuclear leukocytes. To determine if a leukocyte proteinase can modulate plasminogen activation, plasminogen was digested with leukocyte elastase. A major product was a small, approximately 34,000 dalton fragment (mini-plasminogen), without lysine-binding function, but with fibrin-binding activity. After urokinase activation, the resulting mini- plasmin had amidolytic activity for a tripeptide plasmin substrate and fibrinolytic activity. By 125I-fibrin assay, activities of mini-plasmin and plasmin (12 nmole/liter) were 38 and 20 ng fibrin lysed/min, respectively. Lysis times of fibrin clots containing urokinase, and mini-plasminogen or plasminogen (800 nmole/liter), were 282 and 290 sec, respectively. Mini-plasmin and plasmin were inhibited similarly by epsilon-aminocaproic acid and normal plasma, but differed in responses to gel filtration fractions of plasma containing alpha 2-antiplasmin and alpha 2-macroglobulin, the primary and secondary plasmin inhibitors. With purified inhibitors, mini-plasmin required higher concentrations of, or longer preincubation with, alpha 2-antiplasmin, and lower concentrations of, or shorter preincubation with, alpha 2- macroglobulin, to produce inhibition equivalent to that observed with plasmin. Leukocyte elastase digests plasminogen to generate a mini- plasminogen which, when activated by urokinase, has a novel pattern of response to the major plasmin inhibitors in plasma.

scholarly journals Mini-plasminogen: a mechanism for leukocyte modulation of plasminogen activation by urokinase

Blood ◽  
1981 ◽  
Vol 58 (1) ◽  
pp. 97-104 ◽  
Author(s):  
LA Moroz
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Gel Filtration ◽  
Aminocaproic Acid ◽  
Binding Activity ◽  
Major Product ◽  
Plasminogen Activation ◽  
Leukocyte Elastase ◽  
Binding Function ◽  
Plasmin Inhibitors ◽  
Epsilon Aminocaproic Acid

Abstract Urokinase activation of blood fibrinolysis involves polymorphonuclear leukocytes. To determine if a leukocyte proteinase can modulate plasminogen activation, plasminogen was digested with leukocyte elastase. A major product was a small, approximately 34,000 dalton fragment (mini-plasminogen), without lysine-binding function, but with fibrin-binding activity. After urokinase activation, the resulting mini- plasmin had amidolytic activity for a tripeptide plasmin substrate and fibrinolytic activity. By 125I-fibrin assay, activities of mini-plasmin and plasmin (12 nmole/liter) were 38 and 20 ng fibrin lysed/min, respectively. Lysis times of fibrin clots containing urokinase, and mini-plasminogen or plasminogen (800 nmole/liter), were 282 and 290 sec, respectively. Mini-plasmin and plasmin were inhibited similarly by epsilon-aminocaproic acid and normal plasma, but differed in responses to gel filtration fractions of plasma containing alpha 2-antiplasmin and alpha 2-macroglobulin, the primary and secondary plasmin inhibitors. With purified inhibitors, mini-plasmin required higher concentrations of, or longer preincubation with, alpha 2-antiplasmin, and lower concentrations of, or shorter preincubation with, alpha 2- macroglobulin, to produce inhibition equivalent to that observed with plasmin. Leukocyte elastase digests plasminogen to generate a mini- plasminogen which, when activated by urokinase, has a novel pattern of response to the major plasmin inhibitors in plasma.

scholarly journals Fibrin-associated plasminogen activation in alpha 2-plasmin inhibitor deficiency

Blood ◽  
1983 ◽  
Vol 62 (5) ◽  
pp. 1118-1122
Author(s):  
N Aoki ◽  
Y Sakata ◽  
A Ichinose
Keyword(s):  
Latent Period ◽  
Aminocaproic Acid ◽  
Plasminogen Activation ◽  
Amidolytic Activity ◽  
Plasma Clot ◽  
Whole Process ◽  
Congenital Deficiency ◽  
Exponential Increase ◽  
Epsilon Aminocaproic Acid ◽  
Plasmin Inhibitor

The clot formed from the plasma of a patient with congenital deficiency of alpha 2-plasmin inhibitor underwent a spontaneous extensive fibrinolysis. Radiolabeled fibrinogen was added to the plasma before clotting, and the whole process of the fibrinolysis was followed by measuring the release of radiolabels. Plasminogen activation was also followed by measuring the amidolytic activity that developed. There was an initial latent period, followed by an exponential increase of fibrinolytic activity. During the latent period, there was little or no release of radiolabels and no development of amidolytic activity. During the latent period, the clot was washed thoroughly to remove unbound proteins from fibrin and was incubated in buffered saline. The washed clot still underwent fibrinolysis, similar to the original plasma clot, suggesting that the plasminogen/plasminogen activators bound to fibrin during the initial latent period are responsible for fibrinolysis. The amount of plasminogen bound to fibrin during the latent period was close to the amount of plasminogen activated during the whole process of fibrinolysis. When the amount of plasminogen bound to fibrin was decreased by epsilon aminocaproic acid, the extent of fibrinolysis was decreased in parallel with the decrease of the amount of the bound plasminogen. This suggests that the amount of plasminogen bound to fibrin is one of the determinants of the rate of the fibrinolytic process.

scholarly journals Properties of a factor increasing monocytopoiesis (FIM) occurring in serum during the early phase of an inflammatory reaction

Blood ◽  
1977 ◽  
Vol 50 (4) ◽  
pp. 727-742
Author(s):  
D van Waarde ◽  
E Hulsing-Hesselink ◽  
R van Furth
Keyword(s):  
Inflammatory Reaction ◽  
Gel Filtration ◽  
Aminocaproic Acid ◽  
Polystyrene Latex ◽  
Biologically Active ◽  
Clotting Factor ◽  
Half Time ◽  
Epsilon Aminocaproic Acid ◽  
The Complement System

A factor increasing monocytopoiesis (FIM) has been demonstrated during the onset of an acute inflammatory reaction caused by an intraperitoneal injection of polystyrene latex particles. It is protein in nature, does not contain a carbohydrate moiety essential for its function, and is very probably not a glycoprotein. The molecular weight of FIM lies between 18,000 and 24,000 daltons (determined with both ultrafiltration membranes and gel filtration on Sephadex G100). The monocytosis induced by FIM is dose dependent. FIM is thermolabile, having a half-time of about 20 min at 37 degrees C in serum; temperature inactivation can be delayed by the addition of epsilon- aminocaproic acid, the half-time at 37 degrees C then being about 45 min. In vitro treatment of normal murine blood with the inducers of the inflammatory reaction does not result in FIM activity in the serum. FIM dose not have chemotactic activity toward macrophages, is not a clotting factor, is not a biologically active fragment of the complement system, and has no colony-stimulating or-enhancing activity in the vitro bone marrow colony assay. On the basis of these results, a mechanism is postulated for the humoral regulation of monocytopoiesis.

EVALUATION OF PLASMINOGEN IN HYPERCOAGULABLE PATIENTS

1987 ◽  
Author(s):  
R Lottenberg ◽  
J A Hall
Keyword(s):  
Patient Group ◽  
Aminocaproic Acid ◽  
Hydrolysis Rate ◽  
Plasminogen Activation ◽  
Plasmin Activity ◽  
Activation Rate ◽  
History Of ◽  
The Mean ◽  
Epsilon Aminocaproic Acid ◽  
Activation Rates

Sixty seven patients with spontaneous or recurrent thrombosis without obvious underlying disorders were evaluated for plasminogen abnormalities. The mean age was 33.5 years. Fifty seven had venous thrombosis, five had arterial thrombosis and five had both venous and arterial events. Seventeen patients had family members with a history of similar thrombotic events. The plasminogen antigen concentration, by Laurell immunoelectrophoresis, for the patient group was 104± 19% (62-156) compared to 102±11% (83-122) for normal controls (n=24). By amidolytic assay of plasminogen-streptokinase complexes, the activity was proportionate to the antigenic concentration for each individual. Two patients had plasminogen levels (62%, 67%) lower than 2 standard deviations of the mean. Functional alpha2-antiplasmin levels were the same for the patients (102±11%) and the controls (101±16%). Plasminogen activation in undiluted plasma was assessed by a coupled amidolytic assay using urokinase (60 IU/ml) and H-D-val-leu-lys-pNA. The mean substrate hydrolysis rate for the patient group was 1.35±0.52 10−3 absorbance units/min/min (0.56−2.75) compared to 1.32±0.27 10−3 absorbance units/min/min (0.84-−1.99) for the controls (n=34). Three patients with normal plasminogen levels had consistently low activation rates (0.56, 0.57, 0.65). Inhibition of urokinase and plasmin activity was not increased. Histidine-rich glycoprotein levels were not elevated. Electrophoretic, chromatographic and kinetic analysis of the isolated plasminogens revealed no abnormalities. To assess the high affinity lysine binding site, 40 uM epsilon aminocaproic acid was added to plasma while the change in the plasminogen activation rate relative to baseline was observed; no abnormalities were disclosed in either the patient or control populations.In summary, two patients had significantly reduced levels of normally functioning plasminogen. Dysfunctional plasminogen was not identified in this population. We conclude that plasminogen abnormalities infrequently explain hypercoagulability. These studies also suggest that plasminogen status can be adequately evaluated by determining the antigenic concentration and functional activity of plasminogen-streptokinase complexes.

scholarly journals Properties of a factor increasing monocytopoiesis (FIM) occurring in serum during the early phase of an inflammatory reaction

Blood ◽  
1977 ◽  
Vol 50 (4) ◽  
pp. 727-742 ◽  
Author(s):  
D van Waarde ◽  
E Hulsing-Hesselink ◽  
R van Furth
Keyword(s):  
Inflammatory Reaction ◽  
Gel Filtration ◽  
Aminocaproic Acid ◽  
Polystyrene Latex ◽  
Biologically Active ◽  
Clotting Factor ◽  
Half Time ◽  
Epsilon Aminocaproic Acid ◽  
The Complement System

Abstract A factor increasing monocytopoiesis (FIM) has been demonstrated during the onset of an acute inflammatory reaction caused by an intraperitoneal injection of polystyrene latex particles. It is protein in nature, does not contain a carbohydrate moiety essential for its function, and is very probably not a glycoprotein. The molecular weight of FIM lies between 18,000 and 24,000 daltons (determined with both ultrafiltration membranes and gel filtration on Sephadex G100). The monocytosis induced by FIM is dose dependent. FIM is thermolabile, having a half-time of about 20 min at 37 degrees C in serum; temperature inactivation can be delayed by the addition of epsilon- aminocaproic acid, the half-time at 37 degrees C then being about 45 min. In vitro treatment of normal murine blood with the inducers of the inflammatory reaction does not result in FIM activity in the serum. FIM dose not have chemotactic activity toward macrophages, is not a clotting factor, is not a biologically active fragment of the complement system, and has no colony-stimulating or-enhancing activity in the vitro bone marrow colony assay. On the basis of these results, a mechanism is postulated for the humoral regulation of monocytopoiesis.

scholarly journals Binding of fibrin fragments to one-chain and two-chain tissue-type plasminogen activator

Blood ◽  
1992 ◽  
Vol 79 (9) ◽  
pp. 2313-2321 ◽  
Author(s):  
AA Hasan ◽  
WS Chang ◽  
AZ Budzynski
Keyword(s):  
Plasminogen Activator ◽  
Binding Sites ◽  
Solid Phase ◽  
Aminocaproic Acid ◽  
Tissue Type ◽  
Plasminogen Activation ◽  
Tissue Type Plasminogen Activator ◽  
Microtiter Plates ◽  
Type Plasminogen Activator ◽  
Epsilon Aminocaproic Acid

To explore whether fibrin fragments have binding affinity for the tissue-type plasminogen activator (t-PA) molecule, the interactions were studied of (DD)E complex and fragments DD, E1, and E3 with one- chain and two-chain t-PA. For this purpose, a solid-phase binding assay was developed using microtiter plates with nitrocellulose filters. It was found that (DD)E complex and fragments DD and E3 retained the t-PA binding function of the parent fibrin molecule, thus demonstrating that t-PA binds to both the D and E domains of fibrin. Unexpectedly, fragment E1 did not bind t-PA. Fibrin fragments had different binding properties for one-chain and two-chain t-PA. (DD)E complex had the highest and fragment E3 the lowest affinity for one-chain t-PA, both binding curves being consistent with one class of binding sites. However, binding of the fragments with two-chain t-PA was distinguished by more than one class of binding sites, with fragment E3 having the highest affinity for this form of the activator. epsilon-Aminocaproic acid, even at 50 mmol/L concentration, had only minimal effect on binding of (DD)E complex or fragment DD to either one-chain or two- chain t-PA. The potentiating effect of fibrin fragments on plasminogen activation by t-PA was measured by a chromogenic substrate assay. Fragment DD was the most effective stimulator of plasminogen activation by t-PA. In conclusion, (DD)E complex and fragment DD retained most of the regulatory functions of fibrin, which included t-PA binding and t- PA-mediated acceleration of plasminogen activation to plasmin.

scholarly journals Contact activation triggers stimulation of the monocyte 5-lipoxygenase pathway via plasmin

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1941-1951 ◽  
Author(s):  
I Weide ◽  
J Romisch ◽  
T Simmet
Keyword(s):  
Binding Sites ◽  
Polymorphonuclear Leukocytes ◽  
Factor Xii ◽  
Plasminogen Activation ◽  
Contact Activation ◽  
Response Curves ◽  
Lipoxygenase Pathway ◽  
Plasmin Inhibitors ◽  
Pg E2 ◽  
Stimulation Of

The purpose of this study was to characterize the stimulus that activates the 5-lipoxygenase pathway in human peripheral monocytes (PM) during the process of contact activation. Incubation of PM, but not of polymorphonuclear leukocytes (PMN), in contact-activated, recalcified plasma induced a time-dependent release of leukotrienes (LT). The presence of platelets was required for the generation of cysteinyl-LT, but LTB4 formation also proceeded in their absence, although to a lesser extent. Plasmin, presumably generated via the intrinsic fibrinolytic pathway, was liable for the 5-lipoxygenase stimulation during contact activation inasmuch as (1) the 5-lipoxygenase pathway in PM was stimulated by contact-activated, recalcified, autologous or homologous plasma, but not by factor XII-deficient or prekallikrein- deficient plasma; (2) lysine analogs such as N alpha-acetyl-L-lysine, 6- aminohexanoic acid (6-AHA), or trans-4- (aminomethyl)cyclohexane-1- carboxylic acid (t-AMCA), which inhibit plasmin(ogen) binding to PM plasmin(ogen) binding sites, concentration-dependently reduced the cysteinyl-LT release; (3) plasminogen activators such as urokinase or streptokinase concentration-dependently enhanced the cysteinyl-LT release up to 10 and 1,000 IU/mL, respectively, while higher concentrations were less effective leading to bell-shaped concentration- response curves; (4) plasmin inhibitors such as aprotinin or alpha 2- antiplasmin concentration-dependently inhibited the cysteinyl-LT release; and (5) preincubation of plasma with monoclonal antibodies directed against plasminogen and capable of preventing plasminogen activation blocked the contact-mediated 5-lipoxygenase stimulation. Moreover, incubation of PM with plasmin, but not with plasma kallikrein, in Hanks' balanced salt solution (HBSS)-bovine serum albumin (BSA) 0.4% triggered a concentration-dependent release of LTB4 up to 0.1 caseinolytic units (CU)/mL, with higher concentrations being less effective. By contrast, release of cyclooxygenase metabolites such as thromboxane (TX) B2 and prostaglandin (PG) E2 was not stimulated by plasmin, indicating specificity for the 5-lipoxygenase pathway. With plasmin as a hitherto unknown stimulus of the 5-lipoxygenase pathway in PM, a novel link between contact activation and inflammation has been established.

Plasminogen Depletion during Administration of Epsilon-aminocaproic Acid

1973 ◽  
Vol 29 (03) ◽  
pp. 598-602 ◽  
Author(s):  
D. W Nibbelink ◽  
C. D Jacobsen
Keyword(s):  
Subarachnoid Hemorrhage ◽  
Intravenous Administration ◽  
Marked Reduction ◽  
Marked Decrease ◽  
Aminocaproic Acid ◽  
Acute Onset ◽  
Plasminogen Activation ◽  
Quantitative Changes ◽  
Epsilon Aminocaproic Acid

SummarySixteen patients with acute onset subarachnoid hemorrhage were treated with intravenous administration of epsilon-aminocaproic acid. This drug inhibits activation of plasminogen and, to some extent, the effect of plasmin as well. In this study a marked decrease in immunological quantities of plasminogen was observed. This depletion of plasminogen occurred without concommitant quantitative changes in platelets or fibrinogen. Hence, the antiftbrinolytic effect of epsilon-aminocaproic acid is not only through inhibition of plasminogen activation and of plasmin itself, but also through a marked reduction in plasminogen.

A Study of Proteolytic and Fibrinolytic Factors in the Human Prostate

1971 ◽  
Vol 25 (03) ◽  
pp. 481-498 ◽  
Author(s):  
Michelle Nijs ◽  
Christiane Brassinne ◽  
A Coune ◽  
H. J Tagnon
Keyword(s):  
Proteolytic Activity ◽  
Trypsin Inhibitor ◽  
Aminocaproic Acid ◽  
Soybean Trypsin Inhibitor ◽  
Clinical State ◽  
Plasminogen Activation ◽  
The Third ◽  
Human Plasminogen ◽  
Epsilon Aminocaproic Acid

SummaryAn analysis of the proteolytic factors contained in human prostatic tissue was performed in vitro. Casein, fibrinogen and fibrin, non-radioactive and radioiodinated were used as substrates.A first factor, called direct proteolytic activity, capable of proteolyzing casein without prior activation, is described. It had no effect on fibrinogen or fibrin, was inhibited by epsilon aminocaproic acid, but not by the soybean trypsin inhibitor. This shows that this proteolytic activity was quite different from plasmin.A second factor, called plasminogen proactivator, was demonstrated on bovine plasminogen in the presence of streptokinase, the latter being unable to produce direct activation of bovine plasminogen. Activation of this system resulted in the transformation of plasminogen into plasmin, capable of digesting casein as well as fibrinogen and fibrin. Epsilon aminocaproic acid and the soybean trypsin inhibitor inhibited this system. The properties of this proactivator show that it probably does not result from the presence of small amounts of plasminogen in the prostate. Urokinase, a factor present in human urine, is able to activate this proactivator under certain conditions.The third factor, called plasminogen activator, was capable of activating directly human plasminogen into plasmin. It was not active on bovine plasminogen. Epsilon aminocaproic acid and the soybean trypsin inhibitor were effective inhibitors. Addition of large volumes of human prostatic extract to human plasminogen resulted in a paradoxical decrease of the proteolytic activity suggesting the possible existence in the prostate of an inhibitor of this third factor.Possible relationships between these factors and the clinical state of fibrinolysis observed in some cases of disseminated prostatic cancer are discussed.

scholarly journals Contact activation triggers stimulation of the monocyte 5-lipoxygenase pathway via plasmin

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1941-1951 ◽  
Author(s):  
I Weide ◽  
J Romisch ◽  
T Simmet
Keyword(s):  
Binding Sites ◽  
Polymorphonuclear Leukocytes ◽  
Factor Xii ◽  
Plasminogen Activation ◽  
Contact Activation ◽  
Response Curves ◽  
Lipoxygenase Pathway ◽  
Plasmin Inhibitors ◽  
Pg E2 ◽  
Stimulation Of

Abstract The purpose of this study was to characterize the stimulus that activates the 5-lipoxygenase pathway in human peripheral monocytes (PM) during the process of contact activation. Incubation of PM, but not of polymorphonuclear leukocytes (PMN), in contact-activated, recalcified plasma induced a time-dependent release of leukotrienes (LT). The presence of platelets was required for the generation of cysteinyl-LT, but LTB4 formation also proceeded in their absence, although to a lesser extent. Plasmin, presumably generated via the intrinsic fibrinolytic pathway, was liable for the 5-lipoxygenase stimulation during contact activation inasmuch as (1) the 5-lipoxygenase pathway in PM was stimulated by contact-activated, recalcified, autologous or homologous plasma, but not by factor XII-deficient or prekallikrein- deficient plasma; (2) lysine analogs such as N alpha-acetyl-L-lysine, 6- aminohexanoic acid (6-AHA), or trans-4- (aminomethyl)cyclohexane-1- carboxylic acid (t-AMCA), which inhibit plasmin(ogen) binding to PM plasmin(ogen) binding sites, concentration-dependently reduced the cysteinyl-LT release; (3) plasminogen activators such as urokinase or streptokinase concentration-dependently enhanced the cysteinyl-LT release up to 10 and 1,000 IU/mL, respectively, while higher concentrations were less effective leading to bell-shaped concentration- response curves; (4) plasmin inhibitors such as aprotinin or alpha 2- antiplasmin concentration-dependently inhibited the cysteinyl-LT release; and (5) preincubation of plasma with monoclonal antibodies directed against plasminogen and capable of preventing plasminogen activation blocked the contact-mediated 5-lipoxygenase stimulation. Moreover, incubation of PM with plasmin, but not with plasma kallikrein, in Hanks' balanced salt solution (HBSS)-bovine serum albumin (BSA) 0.4% triggered a concentration-dependent release of LTB4 up to 0.1 caseinolytic units (CU)/mL, with higher concentrations being less effective. By contrast, release of cyclooxygenase metabolites such as thromboxane (TX) B2 and prostaglandin (PG) E2 was not stimulated by plasmin, indicating specificity for the 5-lipoxygenase pathway. With plasmin as a hitherto unknown stimulus of the 5-lipoxygenase pathway in PM, a novel link between contact activation and inflammation has been established.

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