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Some fibrinolytic parameters, including tissue-type plasminogen activator (t-PA) and t-PA inhibitor activity, have been studied in three groups of pre-eclamptic patients (severe pre-eclampsia, mild pre-eclampsia and chronic hypertension with superimposed severe pre-eclampsia). These parameters were compared to those observed in a group of normal pregnant women at similar gestational age. An increase in t-PA was observed in pre-eclamptic patients as compared with normal pregnant women. The t-PA inhibitor activity was significantly higher in cases of severe pre-eclampsia than in cases of mild pre-eclampsia and normal pregnancy at similar gestational age. Following the addition of purified t-PA to plasma of severe pre-eclamptic patients, a complex with a molecular weight of about 115 000 was detected by a zymographic fibrin technique. When urokinase (90)% of HMW and 10% of LMW) was used, two complexes of about 95 000 and 85 000 were detected. The increased level of t-PA inhibitor activity found in severe pre-eclampsia was partially inhibited by the antisera against PA inhibitor-1 and PA inhibitor-2. The high level of t-PA inhibitor activity might contribute to explain the presence of localised microthrombi in the placenta in severe pre-eclampsia.

The influence of regular physical exercise on fibrinolysis was studied in two groups of patients after myocardial infarction. The two groups were of similar age and living habits. One of the groups took part in a rehabilitative training programme while the other group did not participate in any sports activities. Several fibrinolytic variables, including plasminogen activator inhibitor (PAI) activity and PAI-1 antigen, were studied before and after an ergometric test performed at three different times: at the end of hospitalization (before beginning the rehabilitation programme), three months and six months after myocardial infarction. Our results indicate that by the sixth month fibrinolytic activity, measured by the tissue plasminogen activator (t-PA) capacity, had decreased significantly (p less than 0.025) in the patients who were not participating in the rehabilitation programme whereas it had increased slightly in the patients involved in the rehabilitation programme in comparison with the initial values. It was also observed that while PAI activity remained constant or decreased slightly in patients after six months in the rehabilitative sports programme, these PAI levels increased significantly in patients who were not in the sports programme. Our results indicate that there was a significant decrease in the fibrinolytic capacity of patients who were not participating in the rehabilitation sports programme, while the patients involved in the rehabilitation programme showed a slight improvement in their fibrinolytic activity.

The soluble fibrin monomer complexes (SFMC) in 154 obstetric patients with possible disseminated intravascular coagulation (DIC) were evaluated in SDS polyacrylamide gel electrophoresis (PAGE) after precipitation with β-alanine. Other coagulation tests were performed on these patients. The patients were classified into three groups: A) patients with a clinical history of DIC (6 cases); B) patients with only the analytical alterations of DIC (35 cases); and C) patients who showed pathological obstetric diagnoses but without a clinical nor analitical history of DIC (113 cases). In the three groups, well-defined bands of less electrophoretic mobility than fibrinogen were obtained. A significant increase in the second electrophoretic band was found in group A (5.1 per cent) when compared to group C (0.5 per cent). The second electrophoretic band appeared in greater proportion in the group of patients with an unfavorable clinical evolution.

SummaryThe functional activity and active site of plasmin in full-term newborns have been studied and compared to those in adults in order to investigate the nature of the abnormality found in newborn plasminogen descried in a previous paper.The functional activity of newborn plasminogen measured on chromogenic substrate was approximately 18% that of adult plasminogen when streptokinase was used as an activator and 12% when urokinase was used.Proteolysis of newborn plasminogen by urokinase yielding a two-chain plasmin form occurred normally, but the incorporation of diisopropylphosphorofluoridate into the light chain of newborn plasmin was approximately 23% of that observed in the light chain of adult plasmin.These observations suggest that the abnormality of full-term newborn plasminogen is located in the active site of the molecule.

Plasma samples from normal labour and from abruptio placentae were treated with ϱ-alanine. The precipitate obtained was studied by 4% polyacrylamide gel electrophoresis. Soluble fibrin monomer complexes (SFMC) were higher than in normal plasma, in normal labour and in abruptio placentae. Qualitative variations in the SFMC were detected. In normal labour the SFMC were composed of two bands and in abruptio placentae three bands were observed. The posibility of using these analytic differences in the diagnosis of intravascular coagulation of abruptio placentae is discussed.

SummaryA relapsing clinical syndrome of skin lesions and disseminated intravascular coagulation (DIC) that showed remission with the infusion of fresh frozen plasma is described in a newborn infant with homozygous deficiency of protein C antigen.This patient presented since birth a recurrent clinical picture of DIC and ecchymotic skin lesions that resembled typical ecchymosis except for the fact that they showed immediate improvement with the administration of fresh frozen plasma. Using an enzyme linked immunosorbent assay method, the determination of protein C antigen levels in the patient, without ingestion of coumarin drugs, showed very low values (No other deficiencies in the vitamin-K-dependent factors or in anti thrombin III, antiplasmin, and plasminogen were found. Seven relatives of the infant had heterozygous deficiency in protein C antigen (values between 40-55%), without clinical history of venous thrombosis. The pedigree analysis of this family suggests an autosomal recessive pattern of inheritance for the clinical phenotype, although an autosomal dominant pattern has been postulated until now in other reported families.We conclude that our patient has a homozygous deficiency in protein C and this homozygous state may be compatible with survival beyond the neonatal period.

A family with inherited factor VII deficiency is described. The propositus is a 9-year-old girl with chronic haemorrhagic history of epistaxis and bleeding after tooth extractions. Her factor VII coag

SummaryVarious parameters of the fibrinolytic system and antigenic and functional protein C and its inhibitor were studied during normal pregnancy and in patients with preeclampsia.The fast acting tissue-type plasminogen activator inhibitor level was found to increase progressively during normal pregnancy. This increase was more evident in cases of severe preeclampsia (p

Il existe une correlation entre l'activite de l'inhibiteur de l'activateur du plasminogene (PAI) et l'antigene PAI-1 chez les patients ayant une thrombose veineuse et/ou un embolisme pulmonaire. L'etude des familles de 5 patients montre une augmentation de l'activite de PAI et PAI-1 chez 3 familles et un desordre thrombolytique chez 1 famille

We studied functional protein C activity, both anticoagulant and amidolytic, as well as protein C antigen in 30 normal subjects, several members of a family with congenital protein C deficiency, 18 patients with severe preeclampsia, 27 patients with coronary heart disease, including 15 patients with myocardial infarction and 12 with angina pectoris, 20 patients on stable oral anticoagulant therapy (thrombotest values: 3-12%) and three patients with disseminated intravascular coagulation. Protein C values measured by the coagulant assay were compared to those obtained with amidolytic and immunochemical assays. In all the groups studied, the activity assays (amidolytic and coagulant) correlated significantly with each other as well as with the immunochemical assay. In patients on oral anticoagulant therapy the coagulant assay gave lower protein C values than amidolytic and immunochemical assays. A good correlation was found between immunological and amidolytic protein C assays (r=0.90, p less than 0.001), immunological and coagulant protein C assays (r=0.93, p less than 0.001), and amidolytic and coagulant protein C assays (r=0.95, p less than 0.001) in all the samples studied without including the protein C values of patients on oral anticoagulant therapy. These results allow us to recommend the functional protein C coagulant assay in patients on stable oral anticoagulant therapy because only this assay evaluates the "in vivo" protein C function in these patients.

After the administration of cytostatic drugs, an increase in thromboembolic phenomena has been described in cancer patients. The authors studied the changes in plasmatic coagulation and fibrinolysis in 40 patients with nonoperable Stage III and IV lung cancer after cytostatic chemotherapy. The results show significant postchemotherapy increases in fibrinopeptide A levels, as well as a decrease in fibrinolytic activity reflected by a drop in functional tissue activator. Also the authors studied the potential accumulative effect of three chemotherapy cycles. A significant increase in functional plasminogen activator inhibitor has been noted. Chemotherapy is apparently capable of conditioning a decrease in fibrinolytic activity in these cancer patients that could be related to the enhanced tendency to developing thromboembolic phenomena after cytostatic chemotherapy.

This report defines the nature of the molecules responsible for the increased plasma plasminogen activator inhibitor (PAI) activity in preeclamptic patients and the relationship of these inhibitors to the severity of placental damage in preeclampsia. Clinical groups consisting of pregnant women with either severe preeclampsia or chronic hypertension with superimposed severe preeclampsia, as well as normal pregnant and nonpregnant women, were analyzed in a panel of functional and immunologic assays for PAI-1 and PAI-2. Pure severe preeclamptic patients in their third trimester showed a significant increase in both antigenic (136 ng/mL) and functional (5.76 U/mL) type 1 PAI (PAI-1) as compared with normal third-trimester pregnant women (34.8 ng/mL and 2.57 U/mL, respectively). In contrast, antigenic (186 ng/mL) and functional (5.76 U/mL) levels of type 2 PAI (PAI-2) were significantly lower in the pure severe preeclampsia group as compared with the values of the normal pregnant group (269 ng/mL and 9.58 U/mL, respectively). The patients with chronic hypertension and superimposed severe preeclampsia exhibited PAI-2 levels comparable to those of the pure preeclamptic group, whereas their antigenic and functional PAI-1 levels were intermediate (94 ng/mL and 3.25 U/mL, respectively) between the normal pregnant and the pure preeclamptic groups. During early puerperium of both normal pregnant women and patients, plasma PAI-1 antigen and activity decreased within one day to approximately the levels detected in normal nonpregnant women, while PAI-2 levels remained elevated for over 11 days. Similar results were obtained in plasma samples obtained from citrated blood and blood collected with an anticoagulant/antiplatelet mixture, suggesting that increased PAI-1 levels in preeclamptic patients were not due to platelet activation in vitro. In preeclamptic patients, a positive correlation between birth weight and PAI-2 values was observed (r = .64, P less than .05), whereas birth weight was inversely correlated with both PAI-1 levels and total PAI activity (r = -.6, P less than .005 and r = -.76, P less than .005 respectively). Preeclamptic patients with extensive placental infarction exhibited higher plasma PAI activity (24.1 U/mL v 11.6 U/mL) and PAI-1 values (305 ng/mL v 80.9 ng/mL) than preeclamptic patients without extensive placental infarction. In contrast, PAI-2 levels were reduced in preeclamptic patients with infarction in comparison with those of patients without infarction (141 ng/mL v 212.9 ng/mL). Our data indicate that increases in the level of PAI-1 accounts for the high plasma PAI activity in severe preeclampsia as measured using single-chain t-PA.

SummaryThe amniotic fluid (AF) when incubated with the patient’s own plasma diminishes the lytic activity of the plasma. It is suggested that this inhibition is due to the presence of fibrinolytic inhibitors in the AF. The inhibitors rate increases as pregnancy advances. Evaluating these inhibitors in a group of 65 women before and after the 38th week of pregnancy, a higher rate of fibrinolytic inhibitors is found after the 38th week. The said differences are statistically significant. For the moment it does not seem that the increasing of the inhibitors in the last part of pregnancy might be used as a fetal maturity test.

Alterations in the coagulation system are common in patients with liver disease. We have examined the importance of the species and chains of fibrinogen in 3 groups of cirrhotic patients. The study of the gelation of fibrinogen in cirrhotic patients shows that the lag time increases in 80.3% of them and that the maximum gelation rate is altered in 51% of these plasmas. Also it is observed that 80% of the plasmas from cirrhotic patients have a percentage (23.3 ± 7.7%) of unpolymerized α chain, after highly cross-linked fibrin formation. These alterations, in lag time and in the maximum gelation rate, have no significant correlation with the situation of the fibrinolytic system in these patients.The study of isolated fibrinogen from cirrhotic patients and normal subjects plasma, shows that there are no objetive alterations in the percentage of fibrinogen species, the amount of sialic acid or the ratio of polypeptide chains.

Protein C, protein S and antithrombin III were evaluated in normal pregnancy, severe preeclampsia and chronic hypertension with superimposed severe preeclampsia. The same study was performed on a group of 10 normal women using oral contraceptives. In normal pregnancy a significant decrease in the level of free and total PS was observed in the 2nd trimester of pregnancy and was sustained throughout the remaining months. No significant changes in the levels of protein C and antithrombin III were observed during normal pregnancy. In preeclamptic states a significant decrease in protein C was observed. It was more evident in severe preeclampsia when compared with the normal pregnancy group at similar gestational age. No statistically significant differences in protein S were found when the normal and pathological groups were compared. Antithrombin III decreased only slightly in the severe preeclamptic group. The decrease in protein C and antithrombin III levels in severe preeclampsia could be related with the microthrombotic state that these patients may present. However, the role played by protein S, which decreases during normal pregnancy and in preeclampsia, is not clear. A decrease in the level of total protein S was observed in the group of women using oral contraceptives. No significant changes in protein C and antithrombin III levels were observed in this group.

Fibrinolysis may be impaired in coronary heart disease patients. 20 coronary heart disease patients and 10 control subjects were examined for tissue-plasminogen activator activity, tissue-plasminogen activator antigen, fast tissue-plasminogen activator inhibitor and other fibrinolytic and haemostatic parameters including antigenic and functional protein C. Both patient and control groups were similar in age and smoking habits. All of these patients had a myocardial infarction between 1-3 months before this study. Assays were evaluated before and after an exercise test. Prothrombin time, activated partial thromboplastin time, protein C, plasminogen, alpha 2-antiplasmin, fibrinogen/fibrin degradation products and contact-activated fibrinolysis were similar before and after exercise in both groups. Fibrinolytic activity assayed by the euglobulin lysis time and fibrin-plate lysis methods was decreased in the patient group as compared with the control group but the difference was not significant. In basal conditions, tissue-plasminogen activator activity was defective in 50% of the coronary heart disease patients (p less than 0.01) and after exercise this percentage rose to 77% (p less than 0.01). However, tissue-plasminogen activator antigen in the coronary heart disease group was similar to that of the control group, both before and after exercise. The activity of the tissue-plasminogen activator inhibitor was persistently increased in coronary heart disease though this increase was not statistically significant. It is concluded that in coronary heart disease patients there is a defective fibrinolytic activity probably due to an increase in tissue-plasminogen activator inhibitor.

Several fibrinolytic variables, including plasminogen activator inhibitor activity, were studied before and after exercise in 67 normolipidaemic patients with coronary artery disease and in 25 hyperlipidaemic patients with coronary artery disease. Before exercise plasminogen activator inhibitor activity was higher in the patient groups than in a group of 10 healthy volunteers. For those who were normolipidaemic plasminogen activator inhibitor activity was greater in patients with angina pectoris who had had a myocardial infarction. The concentration of antigenic tissue-type plasminogen activator was similar in all the patients with coronary artery disease and higher than in the control group. After the exercise test fibrinolytic capacity was lower in the patients with angina pectoris and a previous history of myocardial infarction. After exercise both the released immunological tissue-type plasminogen activator and fibrinolytic capacity were lower in the hyperlipidaemic patients than in the normolipidaemic patients. The concentration of plasminogen activator inhibitor was also higher in the hyperlipidaemic patients. Patients with hyperlipidaemia IV had the highest plasminogen activator inhibitor activity. The increase in plasminogen activator inhibitor activity found in the patients was partially inhibited by antiserum against plasminogen activator inhibitor-1 in vitro. The formation of a complex of about 115,000 daltons between plasminogen activator inhibitor and purified tissue-type plasminogen activator was detected by a zymographic fibrin technique. These findings show that in patients with coronary artery disease fibrinolytic activity is impaired by an increase in plasminogen activator inhibitor. Impaired fibrinolysis may be related to the clinical evolution of coronary artery disease in these patients.

Several parameters of hemostasis have been studied in 19 patients suffering from abruptio placentae. In 10 of them severe hemostatic alterations were detected and in 5, disseminated intravascular coagulation was observed. The patients were divided into four groups according to the severity of their clinical picture. The degree of placental separation was related to the severity and course of the clinical history and to the alterations in hemostasis detected at the most critical clinical moment. The analytical parameters were evaluated after extraction of the thromboplastic material. A good correlation was observed between the severity of the clinical picture and the degree of placental separation and the greatest analytical alteration, especially with cross-linked soluble fibrin monomer complexes (SFMC). In 9 of the 19 patients who showed analytical and/or clinical alterations, an improvement was detected in these alterations after evacuation of the uterus.

SummaryTotal and free protein S antigen and C4b-binding protein (C4bp) were determined by rocket immuno-electrophoresis, and functional protein S was assayed by a coagulation method, throughout pregnancy and normal puerperium and in a group of normal full-term newborns (FTN). The functional protein S assay is based on a modification of the APTT, using a mixture of test sample, protein S deficient plasma, activated protein C, phospholipids and calcium. This protein S functional assay is specific for protein S since the APTT prolongation by normal plasma was abolished by incubation of plasma with monospecific, rabbit antiprotein S IgG. The ratios of functional protein S/free protein S antigen in healthy men (n = 13) and women (n = 14) were 1.0 ± 0.13 (mean ± SD) and 1.03 ± 0.20, respectively. During pregnancy there is a decrease in functional protein S and a progressive decrease in total and free protein S antigen, with a functional/free protein S ratio of 0.75 ± 0.28 in the third trimester of pregnancy (n = 16). In early puerperium the functional protein S level was lower than the free protein S antigen level (ratio about 0.5). In the FTN group, the free protein S level was 39% and protein S activity was about 70% that of adults, with a functional/free protein S ratio of 1.84 ± 0.31. C4bp values were 23.5 ± 10.3% in the FTN group, and crossed immunoelectrophoresis showed that in this group the major protein S peak corresponded to free protein S. These results indicate that both in early puerperium and in FTN group, free protein S antigen may not be an adequate parameter for estimating of functional protein S activity. The decrease in functional protein S activity during early puerperium may be connected with the risk of developing thrombotic episodes during the postpartum period.

The fibrinolytic system was evaluated in a patient with homozygous protein C deficiency as well as in several members of his family with a partial deficiency of this protein. Before anticoagulant therapy the patient showed skin lesions which quickly disappeared after administration of fresh plasma. After anticoagulant treatment, the propositus suffered two clinical episodes of "ecchymotic" lesions, which were controlled with fresh plasma. The patient has remained free of new lesions and other clinical episodes up to the present date. The fibrinolytic activity of both the propositus and his family was normal. The patient's father showed adequate release of tissue plasminogen activator after controlled physical exercise. According to clinical and analytical data from our patient and his family, it is suggested that, in spite of the preservation of the fibrinolytic system in this case, a localized deficiency in fibrinolysis could exist in view of the clinical behaviour of the skin lesions described.

Plasminogen activator inhibitor (PAI) activity and other fibrinolytic parameters have been studied in three groups of preeclamptic patients: A) severe preeclampsia, B) mild preeclampsia and C) chronic hypertension with superimposed severe preeclanpsia. These parameters were compared to those observed in a group of normal pregnant women at similar gestation age ( 3rd trimester of pregnancy, group D). The PAI activity, regardless of whether single-chain t-PA or two-chain t-PA was used, was significantly higher in cases of severe preeclanpsia than in cases of mild preeclanpsia and normal pregnancyThis increase in PAI activity normalizes in early puerperium. When PAI activity was evaluated in the platelet lysate no increase in severe preeclanpsia, as compared to the control group, was observed. Following the addition of purified two chain t-PA to plasma of severe preeclanptic patients, a complex with a molecular weight of about 115,000 was detected by a zymographic fibrin technique. When urokinase (90% of HMW and 10% of LMW) was used, two complexes of about 95,000 and 85,000 were detected. The increased level of PAI activity found in severe preeclanpsia was partially inhibited by the antisera against PAI-1 and PAI-2.

Hemostatic parameters, especially soluble fibrin monomer complexes, have been studied in 114 patients with intrauterine fetal death. The patients were classified into three groups according to the duration of fetal retention. A correlation was observed between a longer duration of fetal retention and the hemostatic alteration. Moreover, in the groups of patients in which this hemostatic disorder was more evident, an improvement in these alterations was detected after fetal evacuation. A hemorrhagic picture with consumption of coagulation factors and cross-linked fibrin oligomers was detected after uterine evacuation in only 1 case and required an emergency hysterectomy and the administration of blood and plasma. In the rest of the patients, fetal evacuation was sufficient to normalize the parameters and no hemorrhagic complications were observed. The favorable evolution and minimal hemostatic alterations in the majority of the patients with intrauterine fetal death, when early obstetric management was established, are discussed.

SummaryA family with “in vitro” increased red-cell fall out from the blood clot was studied. One member of the family (JVM) had a clinical history of hemorrhages after minor trauma or dental extractions. Routine coagulation and platelet function were normal except for the fibrinogen level which was slightly low in several members. The antigenic as well as functional evaluation of factor XIII was within normal limits. No factor XIII inhibitors were present. An increase in the clot permeability index was observed in most family members.The study of the fibrinolytic system showed an enhanced lysis of euglobulins, a normal plasminogen value, normal level of fibrin/ogen degradation products, normal fibrinolytic inhibitors, and an increase in the activity of the plasminogen activator. The activity of this activator was inhibited by an antiserum against tissue-type plasminogen activator. The t-pA inhibitor was in the normal range.It is concluded that the family studied in this paper shows familial alteration in the fibrinolytic system due to an excess of plasminogen activator immunologically related to that in human tissue.