A Plasma Clot Lysis Assay Based on the Release of Fibrin Degradation Products: Application to the Diagnosis of Hypofibrinolytic States

1990 ◽  
Vol 63 (01) ◽  
pp. 076-081 ◽  
Author(s):  
Pascale Gaussem ◽  
Sophie Gandrille ◽  
Pascale Molho-Sabatier ◽  
Loïc Capron ◽  
Jean-Noël Fiessinger ◽  
...  
Keyword(s):  
Degradation Products ◽  
Venous Occlusion ◽  
Lower Limbs ◽  
Clot Lysis ◽  
Plasma Clot ◽  
Vein Thrombosis ◽  
Fibrin Degradation Products ◽  
Before And After ◽  
Euglobulin Clot Lysis Time ◽  
Pai 1

SummaryUsing a monoclonal antibody-based assay, we measured the fibrin degradation product release in the supernatant of plasma clots obtained before and after venous occlusion (VO) in 30 patients with definite or suspected vascular thrombosis (19 definite and 2 suspected deep vein thrombosis, 6 recurrent superficial thrombophlebitis, 3 arterial occlusions of lower limbs). tPA and PAI-1 concentrations were determined using ELISA assays; the post-occlusion values were corrected for haemoconcentration. The increase in tPA during VO was correlated with haemoconcentration (r = 0.74), but 3 patients had ineffective VO (<2% increase in proteins). The fibrinolytic response to VO was evaluated using the shortening of the time necessary for the release of 200 μg of fibrin degradation products per mg of fibrinogen (Δ T 200). Two among the 27 patients with effective VO were bad responders with a Δ T 200 <3 h (whereas all the others had Δ T 200 >10 h). These patients had respectively a deficient tPA release (Δ tPA = 1 ng/ml) and an elevated PAI-1 level at rest (33 ng/ml). Several other patients were bad responders in terms of tPA release or of shortening of the euglobulin clot lysis time but they had a normal Δ T 200. This plasma clot test reflects the ability of free tPA to bind to fibrin (the amount of which depends on the level of tPA and PAI-1), and may be useful in the diagnosis of a hypofibrinolytic state.

Effects of exercise and conditioning on clotting and fibrinolytic activity in men

1987 ◽  
Vol 62 (4) ◽  
pp. 1416-1421 ◽  
Author(s):  
E. W. Ferguson ◽  
L. L. Bernier ◽  
G. R. Banta ◽  
J. Yu-Yahiro ◽  
E. B. Schoomaker
Keyword(s):  
Fibrinolytic Activity ◽  
Blood Clotting ◽  
Degradation Products ◽  
Clot Lysis ◽  
Plasma Clot ◽  
Fitness Program ◽  
Fibrin Degradation Products ◽  
Before And After ◽  
Plasmin Inhibitor ◽  
Sedentary Subjects

Sixty healthy men in three physical fitness categories (sedentary, on no organized fitness program; joggers, running 5–15 miles/wk; and marathoners, running greater than 50 miles/wk) were evaluated for changes in blood clotting and fibrinolytic activity before and after maximum exercise on a treadmill according to the Bruce protocol. The rate of blood clotting, as measured by prothrombin times, partial thromboplastin times and thrombin times, was accelerated by exercise (all P less than 0.005). The ability of euglobulin clots and plasma clots to lyse incorporated 125I-fibrin, termed 125I-euglobulin clot lysis (IEL) and 125I-plasma clot lysis (IPCL), were used as indexes of fibrinolytic activity. Marathoners had greater increases in fibrinolytic activity with exercise (76% compared with 63% for joggers and 55% for sedentary subjects by IEL; 427% compared with 418% for joggers and 309% for sedentary subjects by IPCL; all P less than 0.05). Fibrin degradation products increased with exercise (P less than 0.005 for the total group of 60 subjects). The absolute concentrations of alpha 2-plasmin inhibitor, alpha 2-macroglobulin, and antithrombin III increased with exercise (all P less than 0.005), but when concentrations were corrected for acute shifts of plasma water during exercise, the quantity of these inhibitors actually decreased (all P less than 0.005). The changes in clotting assays with exercise were not significantly correlated with changes in whole blood lactate, blood pyruvate, or rectal temperatures. Fibrinolytic assays before and after exercise correlated poorly to moderately with blood lactates (IEL: r = 0.441 and r = 0.425, respectively; IPCL: r = 0.294 and r = 0.544, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in Clotting and Fibrinolytic Parameters Produced by Venous Stasis

1996 ◽  
Vol 2 (1) ◽  
pp. 64-68
Author(s):  
Raul Altman ◽  
Alejandra Scazziota ◽  
Jorge Rouvier
Keyword(s):  
Sodium Citrate ◽  
Vascular Endothelial Cells ◽  
Protein S ◽  
Venous Occlusion ◽  
Venous Stasis ◽  
Clot Lysis ◽  
Before And After ◽  
Fibrinolytic Parameters ◽  
Pai 1

Vascular endothelial cells have thrombosis- prevention properties through the release of some fibri nolytic factors. This capacity of endothelium was studied using the venous occlusion test (VOT) and measuring proteins released during stasis. Blood samples were col lected before and after 10 and 20 min VOT in tubes con taining sodium citrate at pH 7.0 or 4.3. A significant shortening of the euglobulin clot lysis time (ECLT) was obtained 10 and 20 min after VOT in plasma collected in citrate pH 7.0 or 4.3. No statistical difference was noted between results obtained 10 and 20 minutes after VOT. The shortening was related to the increase of tissue plas minogen activator (t-PA) released during VOT. No differ ence was detected in the concentration of tissue plasmin ogen activator inhibitor 1 (PAI-1), but PAI-1 activity mea sured in plasma from blood collected in citrate pH 7.0 decreased progressively from 10.8 ± 9.2 arbitrary units (AU)/ml to 3.76 ± 4.5 AU/ml and 0.8 ± 1.02 AU/ml (p < 0.001) after 10 and 20 min, respectively, of stasis. When the PAI-1 activity was determined in blood collected in citrate pH 4.3 for preventing in vitro complexing of PAI-1 and t-PA, results were different: PAI-1 activity decreased from a basal value of 29.2 ± 15.3 AU/ml to 24.9 ± 12.2 ( p = NS) and 18.7 ± 11.5; p < 0.02) 10 and 20 min after VOT. As result of fibrinolytic activation, other factors were modified: increase of plasminogen activators, de crease of plasminogen, and increase of D-dimer. Fibrin ogen, fibronectin, protein C, protein S, von Willebrand factor, and tissue factor pathway inhibitor concentration remained unchanged after VOT. According to our find ings, VOT can be performed with a stasis of 10 min. Good responders are related to the endothelial capacity for re leasing t-PA and urokinase-type PA (u-PA) and defined as those with an ECLT of <60 min after 10 min VOT when blood was collected in sodium citrate pH 7.0.

Reperfusion After Venous Occlusion Caused Transient Increase in Plasminogen Activator Inhibitor-1 in Systemic Circulation

1998 ◽  
Vol 4 (2) ◽  
pp. 133-137
Author(s):  
Nobuo Nagai ◽  
Tetsumei Urano ◽  
Yumiko Takada ◽  
Akikazu Takada
Keyword(s):  
Plasminogen Activator ◽  
Fibrinolytic Activity ◽  
Plasminogen Activator Inhibitor ◽  
Systemic Circulation ◽  
Venous Occlusion ◽  
Transient Increase ◽  
Clot Lysis ◽  
Plasminogen Activator Inhibitor 1 ◽  
Euglobulin Clot Lysis Time ◽  
Pai 1

From the point of local regulation, we investigated fibrinolytic activity both in local and systemic circulations, using a venous occlusion test as a stimulus of tissue plasmin ogen activator (t-PA) release in human volunteers. Blood samples were taken before, during, and after venous occlusion from the occluded arm as trapped blood and the unoccluded arm as blood of systemic circulation. In these samples, fibri nolytic activity by euglobulin-clot lysis time and related factors in t-PA and plasminogen activator inhibitor-1 (PAI-1) were measured. Venous occlusion increased fibrinolytic activity ac companied by an increase in t-PA without an increase in PAI-1 on the occluded arm. The fibrinolytic activity on the unoc cluded arm did not change. Five minutes after reperfusion, however, a transient increase in PAI-1 and no change of fibri nolytic activity were observed in the unoccluded arm. Transient increase of PAI-1 after reperfusion was also observed in the occluded arm. These results suggested that increased t-PA by venous occlusion was neutralized by released PAI-1 after reperfusion. Key Words: Tissue-plasminogen activator— Euglobulin-clot lysis time—Fibrinolysis.

The Increase of Leg Fibrinolytic Potential After Reduction of Hydrostatic Stimulus

1983 ◽  
Vol 50 (03) ◽  
pp. 731-734 ◽  
Author(s):  
Dušan Keber
Keyword(s):  
Plasminogen Activator ◽  
Venous Occlusion ◽  
Clot Lysis ◽  
The Third ◽  
Lysis Time ◽  
Before And After ◽  
Euglobulin Clot Lysis Time ◽  
The Difference ◽  
Fibrinolytic Potential ◽  
Clot Lysis Time

SummaryEleven patients were studied sequentially from the beginning of recumbency due to trauma up to the complete mobilization. The first blood sampling was performed 12 hr to 4 days after injury, the second after 12 to 33 days of recumbency and the third after one or more months of mobilization. The blood was drawn each time before and after venous occlusion of the arm and the leg for 20 min. Fibrinolytic potential was calculated as the difference between post- and preocclusion values of plasminogen activator activity, measured with the euglobulin clot lysis time (ECLT) and on fibrin plates. The results showed that fibrinolytic potential of legs after the period of recumbency was approaching that of the arms, being ten times higher as measured with ECLT and five times higher as measured on fibrin plates in comparison with the period after mobilization. It was concluded that hydrostatic pressure was the main, if not the only factor responsible for the difference in the content of plasminogen activator in veins of arms and legs and their fibrinolytic potential.

IDENTIFICATION OF A FIBRINOLYTIC DEFECT IN TWO FAMILIES WITH A TENDENCY TO THROMBOSIS

1987 ◽  
Author(s):  
H Ostermann ◽  
S Koenig ◽  
H Pollmann ◽  
U Schmitz-Huebner
Keyword(s):  
Venous Thrombosis ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Venous Occlusion ◽  
Fibrinolytic System ◽  
Tissue Type ◽  
Clot Lysis ◽  
Normal Range ◽  
Before And After ◽  
Euglobulin Clot Lysis Time

We identified two families in whom one member each suffered from deep venous thrombosis and subsequent pulmonary emboli at the age of 15 and 17. We were able to investigate several members of both families with regard to their fibrinolytic system. Blood sampling was done before and after ten minutes of venous occlusion. Parameters measured were euglobulin clot lysis time (ECLT), tissue-type plasminogen activator (t-PA) activity, t-PA concentration and plasminogen activator inhibitor (PAI) activity, besides several other constituents of the coagulation and fibrinolytic system commonly associated with thrombophilia.In the first family six persons could be evaluated. A prolonged ECLT was found in four probands. Three of them had no measurable t-PA activity after stasis, t-PA concentration after stasis was below the normal range in two and borderline in one of them.In the second family 13 members could be invest-gated. Three adults were found to have a prolonged ECLT. Two of these had very low t-PA activity after stasis, with normal increase in t-PA antigen. Their PAI was increased above the normal range. Six children showed no measurable PAI and increased levels of t-PA activity before stasis. After stasis four children had a prolonged ECLT. Two of them had low t-PA antigen levels, while all of them showed normal t-PA activity increases.These findings suggest, that there may be hereditary defects related to activators and inhibitors of the fibrinolytic system in the investigated families. These could possibly be responsible for the occurence of venous thrombosis early in life. However, results in the second family show that not all of the prolonged ECLT values can be explained by changes of t-PA and PAI.

A Pilot Study of Pro-Urokinase in the Treatment of Deep Vein Thrombosis

1994 ◽  
Vol 72 (03) ◽  
pp. 430-433 ◽  
Author(s):  
Marco Mola ◽  
Pier Mannuccio Mannucci ◽  
Marlo Pini ◽  
Paolo Prandoni ◽  
Victor Gurewich
Keyword(s):  
Deep Vein Thrombosis ◽  
Pilot Study ◽  
Thrombolytic Agent ◽  
Degradation Products ◽  
Lower Limbs ◽  
Vein Thrombosis ◽  
Hemorrhagic Event ◽  
Before And After ◽  
Baseline Values ◽  
Deep Vein

SummarySafety and efficacy of the thrombolytic agent pro-urokinase (pro-UK) in the treatment of deep vein thrombosis of the lower limbs (DVT) have been investigated in an open, uncontrolled, pilot study. Fifteen patients were infused with 800.000 IU (5 mg)/h of pro-UK over 24 h (120 mg), together with unfractionated heparin adjusted to maintain the activated partial thromboplastin time between 1.5 and 2.5 times the basal value. Efficacy was assessed comparing venographic changes in the 11 evaluable limbs before and after pro-UK infusion. The Marder score decreased from a median pre-thrombolysis value of 28 (range 4-40) to 16 (3-38) (p <0.05). One major hemorrhagic event (retroperitoneal bleeding 4 days after the end of the pro-UK infusion) occurred. Fibrinogen, alpha2-antiplasmin and plasminogen significantly decreased from baseline values after 12 and 24 h, fibrin(ogen) degradation products significantly increased. Changes in hemostasis parameters were unrelated to thrombolytic efficacy. The results of this pilot study indicate that pro-UK is thrombolytic in DVT and that it can be administered simultaneously with conventional heparin treatment.

Veränderungen des fibrinolytischen Systems bei Patienten mit peripheren arteriellen Durchblutungsstörungen

VASA ◽  
1999 ◽  
Vol 28 (2) ◽  
pp. 106-111 ◽  
Author(s):  
Poredos ◽  
Stegnar ◽  
Gacnik
Keyword(s):  
Clinical Stage ◽  
Arterial Disease ◽  
Venous Occlusion ◽  
Fibrinolytic System ◽  
Lower Limbs ◽  
Tissue Type ◽  
Clot Lysis ◽  
Before And After ◽  
Atherosclerotic Process ◽  
Peripheral Arterial

Background: The fibrinolytic system may play an important role in the development and progression of peripheral arterial occlusive disease. Patients and methods: The fibrinolytic system of the whole blood and a diseased leg was investigated in twenty men with chronic peripheral atherosclerotic occlusive arterial disease (PAOD, clinical stage II according to Fontaine), aged from 46 to 66 years (mean age = 55.3 years). The diagnosis of PAOD was established by clinical examination and segmental systolic blood pressure measurements using a Doppler ultrasound detector. Twenty age-matched (mean age = 53.4 years) male volunteers with normal arterial circulation of the lower limbs and without risk factors of atherosclerosis, served as controls. In both groups fibrinolytic system was investigated in basal conditions and during provocation. Release of tissue-type plasminogen activator (t-PA) was provoked by 20 min venous occlusion of the arm and the leg and by infusion of DDAVP (1-desamino-8-D-arginine-vasopressin, 0.4 ug/kg of body weight). Blood samples were obtained from the arm and the leg before and after each stimulus. The fibrinolytic parameters: euglobulin clot lysis time, t-PA activity (amidolytic assay) and antigen (ELISA) and t-PA inhibitor (PAI) activity (amidolytic assay) were determined. Results: With the exception of a boderline increase in PAI activity in patients, no other differences between the two groups were observed in basal conditions. The most prominent deterioration of the fibrinolytic system detected in male PAOD patients was a significantly higher residual PAI activity registered during venous occlusion of the arm and two minutes after combined stimulation. Two minutes after combined stimulation (DDAVP and venous occlusion of the arm) significantly lower t-PA activity was observed in patients. In patients t-PA antigen response to venous occlusion and DDAVP was not significantly different from the response observed in healthy volunteers. The fibrinolytic response of the leg to venous occlusion was poor and after DDAVP application it was comparable to the arm. The fibrinolytic response of the diseased leg in men was not significantly different from the healthy leg. Conclusion: The results of our study indicate that alteration of the fibrinolytic system in atherosclerotic disease is predominantly a generalised phenomenon and is not directly related to a local atherosclerotic process.

The Fibrinolytic Response To Stanozolol In Normal Subjects

1981 ◽  
Author(s):  
M Greaves ◽  
F E Preston
Keyword(s):  
Degradation Products ◽  
White Cell Count ◽  
Normal Subjects ◽  
Clot Lysis ◽  
Base Line ◽  
Blood Samples ◽  
Fibrin Degradation Products ◽  
Euglobulin Clot Lysis Time ◽  
Group A ◽  
Group B

It has been recognized for a number of years that fibrinolytic activity in patients with various forms of vascular disease may be enhanced by stanozolol (17-β-hydroxy-17α-methyl adrostano {3.2,- c } pyrazole ). However, little is known of its effect in normal healthy individuals. Also it is not known how quickly the drug exerts its effects. Two groups of eight normal healthy adults were studied. Seven subjects were included in both groups. Group A received stanozolol 5mgs b.d. Group B received 5mgs daily. Before receiving the drug, two base line studies were performed on blood samples from each individual. Thereafter, blood samples were taken at 2,7, 10, 17, 24, 31 and 38 days after commencement of treatment. The following investigations were performed - haemoglobin, white cell count, platelet count, euglobulin clot lysis time (ECLT), fibrinogen (Fg), plasminogen (Pg), α2 macroglobulin (α2M), fibrin degradation products, serum albumin, liver enzymes. In Group A statistically significant increases in plasminogen activator activity, measured by the ECLT, and Pg were observed 2 days (P<0.05) and 10 days (P<0.05) respectively after administration of stanozolol; these remained elevated throughout the study. Also, significant reductions of plasma Fg (P<0.01) and α2M (P<0.05) occurred within 7 days of commencement of the drug and these also persisted throughout the six week period of the trial. Similar statistically-significant changes were also observed in Group B subjects in ECLT, Pg, α2M, although the induced changes were less predictable than those observed in Group A. These results indicate that stanozolol exerts significant and persistent enhancement of fibrinolysis in normal individuals.

Hemostatic Disorder of Uremia: The Platelet Defect, Main Determinant of the Prolonged Bleeding Time, Is Correlated with Indices of Activation of Coagulation and Fibrinolysis

1996 ◽  
Vol 76 (03) ◽  
pp. 312-321 ◽  
Author(s):  
Diego Mezzano ◽  
Rodrigo Tagle ◽  
Olga Panes ◽  
Marcos Pérez ◽  
Patricio Downey ◽  
...  
Keyword(s):  
Renal Failure ◽  
Bleeding Time ◽  
Degradation Products ◽  
Plasminogen Activator Inhibitor ◽  
Primary Hemostasis ◽  
Prolonged Bleeding Time ◽  
Fibrin Degradation Products ◽  
Von Willebrand ◽  
Pai 1 ◽  
Coagulation And Fibrinolysis

SummarySeveral parameters of primary hemostasis and markers of activation of coagulation and fibrinolysis were measured in 48 patients with severe (creatinine clearance <20 ml/min) chronic renal failure (CRF) without dialysis and diseases or drugs affecting hemostasis. Bleeding time (BT) was prolonged in 25/48 patients, and was correlated with age of patients, severity of renal failure, hematocrit, impairment in platelet aggregation-secretion and decrease in platelet ATP content. Defects in von Willebrand factor played no role in the prolongation of the BT. Multivariate analysis showed that only platelet dysfunction and severity of renal disease were independent predictors of the BT in uremia. The platelet functional disorder was significantly correlated with a reduction in platelet ATP and ADP.High levels of plasma thrombin-antithrombin complexes (TAT), prothrombin fragment F1+2, fibrinogen and factor VIIc were observed in patients with CRF, as described in prethrombotic states. Plasmin-antiplasmin complexes (PAP), fibrinogen and fibrin degradation products (FgDP, FnDP) were significantly increased, and the activity of plasminogen activator inhibitor (PAI-1) was slightly reduced, denoting an activation of fibrinolysis.A negative correlation was found between platelet levels of ATP and ADP with plasma TAT, F1+2 and PAP. Furthermore, plasma PAI-1 activity was negatively correlated with the BT and was lower in patients with prolonged BT as compared with controls and patients with normal BT. These links between primary hemostasis and activation of coagulation and fibrinolysis suggest that increased intravascular generation of thrombin and/or plasmin is an important mediator of the defects in primary hemostasis, prolongation of the BT and, probably, bleeding in CRF.

PHYSICAL ACTIVITY RELEASES TISSUE PLASMINOGEN ACTIVATOR FROM EXERCISING PARTS OF BODY

1987 ◽  
Author(s):  
I Keber ◽  
K Potisk ◽  
D Keber ◽  
M Stegnar ◽  
N Vene
Keyword(s):  
Physical Activity ◽  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Fibrinolytic Activity ◽  
Venous Occlusion ◽  
Clot Lysis ◽  
Lysis Time ◽  
Tissue Plasminogen ◽  
Euglobulin Clot Lysis Time ◽  
The Difference

To determine the origin of tissue plasminogen activator (t-PA) release during physical activity, we studied the separate and combined effects of venous occlusion and acute physical activity on t-PA release in arm and leg. In 15 healthy volunteers 20 min venous occlusions of arm and leg were performed simultaneously before physical activity ( maximal stress testing on treadmill)(occlusion I), immediately after physical activity and 45 min later (occlusion II). Blood samples were drawn from unoccluded arm before occlusion and after physical activity, and from occluded arm and leg after occlusion. Fibrinolytic activity was measured by euglobulin clot lysis time (ECLT) and t-PA activity assay. The amount of released t-PA during different stimuli (fibrinolytic potential) was calculated as the difference between post- and prestimulation fibrinolytic activity. Before physical activity there was a great increase in fibrinolytic activity due to t-PA in the occluded arm but no increase in the occluded leg. Physical activity itself caused a similar increase of systemic fibrinolytic activity as arm occlusion locally. After physical activity arm occlusion evoked equally good response than before it. Fibrinolytic activity during leg occlusion behaved differently: there was an increase in t-PA activity in the occluded leg which persisted one hour after physical activity, when systemic fibrinolytic activity already fell to initial level.These results demonstrated that walking and running triggered t-PA release from the leg vessels. Since leg occlusion was not a stimulus for t-PA release, it served only as a method to demonstrate the effect of physical activity.

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