Fibrinolytic Tests Research Articles

Effects of Thrombolytic Therapy on Hemorrhage from Post-operative Wounds

SummaryHemorrhage from post-operative wounds in different organs and structures during thrombolytic therapy was investigated in relation to the time after surgical trauma and to the results of laboratory tests of coagulation and fibrinolysis. Streptokinase was given to rabbits at different times following the production of standardized wounds in different organs. Bleeding was frequent and severe when streptokinase was given between one and five days after surgery and was most marked from artery, followed in order by kidney, vein, liver, muscle, gut and skin. Bleeding appeared to be associated with the in vitro demonstration of an active systemic fibrinolytic state together with elevated levels of fibrin degradation products.

CHAPTER XII: Interrelationships of Pulmonary Angiograms, Lung Scans, Hemodynamic Measurements, and Fibrinolytic Findings

Data from pulmonary angiograms, lung scans, hemodynamic measurements, and tests of fibrinolysis were compared and correlated. In general, there was good correlation of the preinfusion scan perfusion defect with the overall angiographic severity, judged both subjectively and objectively. The highest correlation was obtained when only perfusion defects estimated by the two methods were compared. The correlation was lower when 24-hour changes estimated by scanning and angiography were compared. There was a tendency for the angiogram to reflect greater change. Hemodynamic abnormalities did not correlate well with the extent of preinfusion involvement judged by scanning or angiography, although in the urokinase patients there was a correlation of the fall in pulmonary arterial mean pressure with scanning or angiographic improvement. This correlation was not present in the patients treated only with heparin. There was no correlation of degree of fibrinolysis with resolution of emboli as estimated by lung scanning or angiography. This was true in all patients who received urokinase, regardless of initial clot size or degree of change after urokinase infusion.

Comparative study of coagulation, fibrinolysis and cardiorespiratory function in elderly and young subjects after exercise.

ABSTRACT:Nine aged subjects (67 to 87 years old) and 9 young and adult persons were submitted to an exercise load (cycloergometer) varying between 1.0 to 1.5 W/kg. Tests of cardiorespiratory function, blood coagulation and fibrinolysis were carried out before and after exercise, and then after 5, 15 and 60 minutes. The increase of respiratory rate was more marked in the elderly subjects than in the young and adult persons. No differences were observed with respect to the increase of heart rate and blood pressure. The increase of oxygen “pulse” was considerably less pronounced in the elderly. The pulmonary ventilation and the ventilatory oxygen equivalent were significantly different after six minutes of exercise. No significant differences were observed for blood lactic and pyruvic acid, or for the blood coagulation indexes, including factor VIII. The increase in fibrinolytic activity was marked and constant, irrespective of age.

Coagulation, fibrinolysis and platelet function in pre-eclampsia, essential hypertension and placental insufficiency.

SummaryTests of coagulation, fibrinolysis and platelet function were performed in patients with normal pregnancy, severe pre‐eclampsia, mild pre‐eclampsia, essential hypertension and placental insufficiency.Compared with normal pregnancy, severe pre‐eclampsia was associated with higher serum and urinary fibrinolytic degradation products, cryofibrinogen and factor VIII levels, lower plasminogen and antithrombin activity and diminished sensitivity to urokinase‐induced fibrinolysis. In addition severely pre‐eclamptic patients had reduced platelet counts, lower response to adenosine diphosphate platelet aggregation and diminished platelet factor 3 activity. These changes were present before delivery and in the immediate puerperium, but six weeks after delivery the only difference detected between severely pre‐eclamptic and normal patients was reduced platelet disaggregation in the severely pre‐eclamptic group. In mild pre‐eclampsia the changes in coagulation and fibrinolysis were less marked but serum fibrinolytic degradation products, cryofibrinogen, factor VIII and V levels were significantly raised and the platelet count reduced.No abnormalities of coagulation, fibrinolysis or platelet function were detected in essential hypertension. In placental insufficiency there was diminished platelet disaggregation before delivery but all other tests of coagulation, fibrinolysis and platelet function were similar to normal pregnancy.These results support the hypothesis that intravascular coagulation is present in pre‐eclampsia.

EFFECT OF COMBINED ŒSTROGEN-PROGESTOGEN ORAL CONTRACEPTIVES, ŒSTROGEN, AND PROGESTOGEN ON ANTIPLASMIN AND ANTITHROMBIN ACTIVITY

Plasma antiplasmin and serum antithrombin activity together with other tests of coagulation and fibrinolysis were estimated in female volunteers during the normal menstrual cycle and during treatment with oestrogen, progestogen, and œstrogen-progestogen oral contraceptives. Œstrogen and combined œstrogen-progestogen therapy increased plasma antiplasmin and decreased serum antithrombin activity. These changes were accompanied by a rise in plasminogen and a small rise in fibrinogen. The combined preparations also shortened the kaolin partial thromboplastin-time and increased sensitivity to urokinase-induced fibrinolysis. In contrast, no significant changes were seen in coagulation or fibrinolysis during the normal menstrual cycle or progestogen therapy.

Intestinal Alkaline Phosphatase as a Thrombogenic and Platelet-Altering Factor in Rabbits

SummaryPurified intestinal alkaline phosphatase (AlPh), 10 mg/kg, was injected intravenously into rabbits. In normal animals, 1 min post-injection, averaged samples showed platelet count decreases to 50% of control values in whole blood and to 20% in centrifuged “platelet-rich” plasma. Small or large platelet aggregates were noted in the counting chambers, and a glass-bead platelet-adhesiveness test gave values at least twice that in the controls. The platelet changes were paralleled by shortening of an ear-lobe bleeding-time test and by an equally temporary shortening of the r and h values in thrombelastograms (TEG) of fresh whole blood. A very significant correlation, in the 1 min data, was with 4 + thrombosis in 30 min occluded test segments of mesenteric veins. Screening tests of coagulation and fibrinolysis were not significantly altered, and the same was true of various clotting factor assays, except perhaps for small changes which might be attributable to limited local clot formation.15 min post-injection, platelet counts had risen again, no aggregates were noted, the TEG normalized, and the thrombosis test was negative.In dicumarolized rabbits, the AlPh injections still elevated the serum alkaline phosphatase and caused the platelet alterations, but now without the thrombophilic changes.Electrophoretic studies of AlPh isoenzyme distribution suggested that the significant post-injection elevations were not those in the total serum or beta-globvlin enzyme levels, but in a specific “gamma” isoenzyme, migrating cathodic to the ß-globulins.The significance of the present experiments to the basic understanding and testings of thrombotic states is briefly discussed.

The Preparation and Evaluation of a Standardized Fibrin Plate for the Assessment of Fibrinolytic Activity

SummaryA new fibrin plate technic for evaluating components of the fibrinolytic system has been developed. It provides quick, accurate, and easily interpreted results for the fibrinolytic profile. The standardized human plasminogen-free fibrin plates can be produced in bulk and stored for prolonged periods of time. A test specimen placed in a well punched in the buffered agarose gel diffuses into the agar and lyses the fibrin clot, forming a clear reaction zone. The zone diameter is directly proportional to the log of the percent concentration of available fibrinolytic enzyme in the specimen. The plates may be used to quantitate total plasminogen, and estimate available plasmin and active plasmin. A good correlation between results obtained using these fibrin plates and caseinolytic methods was found. Performance and interpretation of tests of fibrinolysis done on these new fibrin plates indicate that it may be the most sensitive technic available for clinical laboratory work.

Coagulation Defects in Acute Promyelocytic Leukemia

Four patients with acute promyelocytic leukemia manifested multiple coagulation abnormalities including hypofibrinogenemia, prolongation of prothrombin and thrombin times, and varying deficiencies of coagulation factors. One of the patients (patient 2) showed rising levels of fibrinogen without changes in other coagulation factors on heparin therapy and demonstrated widespread intravascular coagulation on postmortem examination. Another patient (patient 1) had positive tests for active fibrinolysis and an absolute lack of the usual postmortem thrombi. The current hypotheses of the pathogenesis of the coagulation abnormalities are presented as well as guide to therapeutic management.

A STUDY OF THE FIBRINOLYTIC ACTIVITY IN PULMONARY TUBERCULOSIS

Author studied the fibrinolytic activity of preoperative systemic venous blood and that of systemic and pulmonary venous blood during operation. and reviewed the relationship between the fibrinolytic activity and the clinical findings. in 47 cases of pulmonary resection for pulmonary tuberculosis.The results were as followings: 1. In pulmonary tuberculosis, the preoperative positive cases of fibrinolytic activity test were 32%, and it was significant compared with the other pulmonary lesions. 2. During operation, the fibrinolytic activity of systemic venous blood was moderately positive in 36% of the cases. but that of pulmonary venous blood was moderately or highly positive in all cases. 3. In systemic venous blood, nearly all of the preoperative negative fibrinolytic cases were negative during operation. but in the Pulmonary venous blood, all cases were posítíve.4. The preoperative positive fibrinolytic activity cases in systemic venous blood, were moderately accentuated in fibrinolytic activity during operation but in pulmonary venous blood, all cases were markedly accentuated. 5. It revealed that the extent of the lung parenchyma destruction was proportional to the accentution of the positive fibrinolytic activity.6. In preoperative positive fibrinolytic activity cases, the amount of post-operative abnormal bleeding was significantly increased compared with the preoperatively negative cases.7. It seemed that the preoperative administration of anti-plasmin agents (such as E-ACA) had An effect upon the reduction of the amount of postoperative e bleeding.8. The preoperative fibrinolytic activity test might be very helpful in preventing the postoperative abnormal bleeding.

A Case Report of "Cape Cod" Rocky Mountain Spotted Fever with Multiple Coagulation Disturbances

A case of "Cape Cod" Rocky Mountain Spotted Fever complicated by thrombocytopenia, circulating anticoagulant activity, and deficiency of Vitamin K-dependent clotting factors that responded to the intravenous administration of Vitamin K has been presented. To our knowledge, this is the first such case described. Coagulation studies supported, in part, by grants from the Wimpfheimer Foundation and the John A. Hartford Foundation. We are indebted to Dr. Sherwin V. Kevy of the Children's Hospital Medical Center for performing tests of fibrinolysis and to Dr. Edward S. Murray of the Department of Microbiology of the Harvard School of Public Health for his assistance in the isolation of Rickettsia and the performance of the serologic tests for diagnosis.

Effect of physical exercise on blood clotting and fibrinolysis.

The effect of strenuous exercise on the clotting and fibrinolytic systems was studied on 1 Hageman-deficient and 59 normal subjects (males aged 18–37 years). In the normal subjects there was a significant shortening of the whole-blood clotting time and of the partial thromboplastin time both in glass and in siliconized tubes. Plasma factor VIII (AHF or AHG) assays rose to 188% (average), but the specificity of the test is questioned. Factor XII (HF) increased to 318% (average) unequivocally. A postexercise increased heparin tolerance was also noted. There was no significant increase in the levels of fibrinogen, prothrombin, factor V (AcG), or factor VII (proconvertin) and factor X (Stuart). Fibrinolytic activity as measured by the euglobulin lysis and plasma plate methods increased significantly in most of the normal subjects. The data suggest that the fibrinolytic factor which increases after exercise is not active plasmin, but is related to the “activator” mechanisms. A plasma lysokinase (indirect activator) seems to preponderate in over half the cases. In 20% of cases a plasminoplastin (direct activator) may be involved. In the Hageman-deficient subject there was no improvement in clotting, and the slight changes in some of the fibrinolysis tests were nonsignificant. Submitted on October 16, 1962 Submitted on October 16, 1962

Fibrinolysis as Related to the Urea Solubility of Fibrin

SummaryThe in vitro experiments described in this report demonstrate that the rate of fibrinolysis as determined by both the spontaneous whole blood fibrinolysis test and the fibrin plate procedure is altered by variations in technique. The addition of calcium ion to the fibrinogen before clotting and the heating of fibrin resulted in similar decreases in fibrinolytic activity. Probably the same mechanism was involved, since both the addition of calcium and heating are known to cause the conversion of hydrogen-bonded fibrin to the sulfhydrylbonded form. The finding of both kinds of fibrin in spontaneously clotted whole plasma and in clots taken from the heart postmortem suggests that the type of fibrin bonding may influence the rate of removal of clots in vivo.

A Method for Preservation of Oxalated Plasma Clot for Fibrinolytic Tests

Oxalated plasma usually loses its ability to form a clot upon addition of calcium chloride when kept in the icebox for 72 hours.We have been testing the fibrinolytic power of hemolytic streptococci immediately after isolation and at regular intervals thereafter to observe any changes in this property and its relation to other biological characteristics of the organism. A susceptible plasma clot from an individual was used repeatedly, necessitating a considerable number of bleedings, resulting usually in loss of time and material. For these reasons we decided to search for a method to preserve plasma while retaining its ability to clot readily and maintaining its susceptibility to lysis unaffected. This has been accomplished by the following method:Human blood from a susceptible person was obtained and distributed in 5 cc amounts into small glass bottles, each containing 10 mg of potassium oxalate in the form of the dry powder. After mixing well by gentle shaking, the plasma was separated by centrifugation...

The Fibrinolytic Activity of Hemolytic Streptococci from Normal and Diseased Throats

Since the report in 1933 by Tillett and Garner' of a fibrinolytic capacity in beta hemolytic streptococci isolated from severe human infections, several investigators have suggested that a test for fibrinolytic activity might be employed as a means of indicating the virulence of such strains. This test, if proved to be reliable, would be valuable in the early differential diagnosis of acute and mild infections, in the detection of carriers, in the diagnosis of scarlet fever, in the examination of contacts and in the examination of patients for quarantine release. Accepting the presence of actively fibrinolytic hemolytic streptococci in severe infections to have been adequately demonstrated, we have made throat cultures from a large number of normal persons and from a number of those suffering with mild infections, in order to determine if actively fibrinolytic strains are ever present in the absence of a severe infection. In view of favorable results from this study we have later investigated the application of the test to the control of scarlet fever. For our first study we chose a group of 10 students who were apparently free from respiratory infections. We made 8 cultures from each student, at weekly intervals, and tested the hemolytic and fibrinolytic activities of all of the beta hemolytic streptococci isolated. It was hoped that observations could be made on the fibrinolytic activity of streptococci isolated during the onset, the height and the convalescent stages of minor respiratory infections as well as during the absence of evident infection. Each throat swab was washed off in about 10 ml. of warm, sterile physiological salt solution, and the suspension was plated out on horse blood agar. After 18-24 hours' incubation if hemolytic, streptococcus like colonies were present, 8-10 were picked into horse meat infusion broth. The resultant cultures were further purified by plating out on blood agar and those which did not prove to be streptococci were eliminated after microscopic examination. Transfers were made into serum broth from which hemolysin titrations were made after 24 hours' incubation, and into infusion broth from which, after 18 hours' incubation, fibrinolytic tests were made according to the Tillett and Garner' plasma clot technic. In this, as well as in later studies, 4 strains of streptococci, originally isolated from severe infections, that were lytic in less than 30 minutes, were used as checks in order to avoid false results due to the use of a resistant plasma. In addition, we obtained one culture from each of 123 persons, who were apparently free from respiratory infections, and tested the fibrinolytic and

Fibrinolytic Specificity of B. pestis.

Applying the plasma-clot technic, Tillett and Garner1 demonstrated fibrinolytic enzymes in broth cultures of streptococci and staphylococci. All other bacterial species tested by them were fibrinolytically negative.A negative reaction with the plasma-clot technic, however, is not conclusive evidence of the absence of a fibrinolytic factor. Many plasma-clots are known to contain neutralizing antibodies. We have, therefore, tested all locally available bacterial species (231 strains) by the serum-free fibrin-clot technic.In addition to certain pyogenic cocci, all locally available strains of B. pestis are strongly fibrinolytic, particularly when tested with rat- or guinea pig-fibrin. Fibrinolytic tests with the 16 available bubonic plague cultures are recorded in Tables I and II.Strain 1 in Tables I and II is a 20-year-old stock culture of B. pestis originally isolated from a human case. Strains 2–16 have been isolated during the last 12 months from field mice and ground squirrels. These 15 strains were kin...

Fibrinolytic Activity of Beta Hemolytic Streptococci from Cow's Milk

One of the many unsettled problems concerning mastitis is its relationship to the organism designated as S. mastitidis. While the majority of workers agree that this relationship must be a very significant one, any new approach to the study of this subject should be of interest. Tillet and Garner found that many beta hemolytic streptococci from human diseases were capable of causing specific lysis of human plasma clots. Negative fibrinolytic tests were observed with other bacterial species from human sources, and strains which would actively lyse human plasma clots would not cause the dissolution of rabbit fibrin. Later Tillet stated, “There is a relationship between the infectivity of beta hemolytic streptococci for man and the fibrinolytic property of the cultures.” Madison confirmed the work of Tillet and Garner and further tested strains from horse, rabbit, man, and hog. The lytic action of each strain was determined on fibrin from horse, hog, cow, rabbit, and man, but he did not test organisms from the cow on bovine plasma or fibrin. Thus, no conclusions could be drawn respecting the applicability of these tests to the study of S. mastitidis. If the organism usually associated with infectious mastitis in the cow is able to lyse bovine fibrin, then fibrinolysis may offer a useful method of studying the r6le of this organism in mastitis.

Fibrinolysis by Streptococci of Human and Animal Origin.

The discovery by Tillett and Garner of an antihuman fibrinolytic function in S. hemolyticus isolated from human cases and the absence of this enzyme from apparently identical streptococci isolated from the environment or from lower animals, is of suggestive epidemiologic interest. We have, therefore, attempted to confirm their findings. To do this, 193 cultures of typical hemolytic streptococci were kindly furnished by various clinics and veterinary institutions of Northern California. Antihuman fibrinolytic tests were made by the Tillett-Garner technic, each test being repeated 3 times with different samples of normal human blood. Control tests were made with stock cultures of known lytic powers. For convenience in recording our data, the 193 cultures were divided into 3 groups: (a) 32 cultures originally isolated from internal human tissues, representing such diseases as pneumonia, septicemia, empyemia, endocarditis, osteomyelitis, and meningitis; (b) 123 typical hemolytic strains originally isolated from superficial human tissues, representing such diseases as erysipelas, furunculosis, and fistula, septic sore throat, sinusitis, and acute gastric disturbance; and (c) 38 cultures of veterinary origin, representing such animals as cow, horse, hog, dog, deer, and rabbit. The lytic tests thus grouped are recorded in Table I. Groups I and III are in practical accord with the Tillett-Garner data. Group II, however, suggests a much less sharp differentiation between antihuman and antiveterinary streptococci than their data indicate. The low percentage of fibrinolytic hemolytic streptococci in this group suggests that a fibrinolytic typing of superficial human infections might be of clinical interest. Thirty-three strains of S. viridans were tested in connection with this work. All were non-fibrinolytic.