Antithrombin III activity and concentration in diabetes mellitus.

Antithrombin III (AT III) levels have been reported to be low, normal, and high in diabetes mellitus. Furthermore, a discrepancy between AT III activity and antigen concentration was reported. We have evaluated the behaviour of AT III activity and antigen level both in type 1 and type 2 diabetes, either in uncontrolled or in well controlled patients. AT III activity and antigen levels showed values similar to normal. No difference was seen between type 1 and type 2 diabetes. Similar results were observed in the group of well controlled diabetic patients. AT III activity and antigen did not correlate with blood glucose and glycosylated haemoglobin (HbA1). No difference was observed between AT III activity and antigen levels in any group. Therefore the hypercoagulable state found in diabetes mellitus does not depend on AT III modifications. A discrepancy between AT III activity and antigen was not confirmed. A dysantithrombinaemia, explained on the basis of an inactivation of protein glycosylation in diabetes mellitus has not been confirmed.

The effect of low-dose estroprogestinic preparations on prothrombin complex factors: no significant increase after an 8-month trial.

The behavior of the prothrombin complex factors in 16 healthy women during low-dose estroprogestinic treatment (laevonorgestrel 0.15 mg and ethynilestradiol 0.30 mg) at basal conditions and during 8 months of therapy has been investigated. We found a statistically significant decrease of the PTT (Partial Thromboplastin Time). The prothrombin time, on the other hand, became slightly decreased, but not to a statistically significant extent. Among the prothrombin time derived tests for evaluating the prothrombin complex only the PP test (Prothrombin Proconvertin test) was significantly shortened. Of the coagulation factors (factors II, VII and X) only a modest, but not statistically significant, increase in Factor VII and Factor X was noted. We conclude that, during the 8 month observation period, prothrombin complex factors are not altered substantially.

PIP: The behavior of the prothrombin complex factors in 16 healthy women during low-dose estroprogestinic treatment (levonorgestrel 0.15 mg and ethyinyl estradiol 0.35 mg) at basal conditions and during 8 months of therapy has been investigated. A statistically significant decrease of the partial thromboplastin time (PTT) was found. The prothrombin time, on the other hand, became slightly decreased, but not to a statistically significant extent. Among the protrombin time derived tests for evaluating the protrombin complex only the protrombin proconvertin (PP) test was significantly shortened. Of the coagulation factors (factors II, VII, and X) only a modest, but not statistically significant, increase in Factors VII and Factor X was noted. The authors conclude that, during the 8 month observation period, prothrombin complex factors are not altered substantially.

Behaviour of antithrombin III abnormalities in the crossed immunoelectrophoresis system.

Antithrombin III (AT III) abnormalities can be characterized by means of crossed immunoelectrophoresis. In the past, it was thought that the abnormalities could be demonstrated only if heparin is present in the system. Now some conditions (AT III Trento, for example) are known to show an abnormal pattern only in the absence of heparin. This indicates that some of the changes are heparin-independent. Furthermore, it could be demonstrated that in some cases the abnormality is present only in serum (AT III Vicenza, for example). Therefore, the test should be carried out as a screening procedure both in plasma and serum and in the presence or absence of heparin in every case of suspected AT III abnormality.

Factor X Padua: a 'new' congenital factor X abnormality with a defect only in the extrinsic system.

A family with a new factor X defect is reported. The proposita is a 56-year-old female. She is asymptomatic and no consanguinity is present between the parents. The main features of the defect are: prolongation of prothrombin time and derivative tests but normal partial thromboplastin time. Factor X was found to be low (about 25-30% of normal) only if tissue thromboplastins were used in the assay system. Chromogenic substrate S-2222 also yielded decreased factor X levels. However, factor X activity was normal with cephalin and cephalin-RVV mixture. Factor X antigen was normal in three immunological systems (electroimmunoassay, an Elisa method and laser nephelometry). Crossed immunoelectrophoresis and antigen-antibody kinetics recorded in a laser nephelometer failed to show major differences from normal factor X. Both sons of the proposita, the father and other family members showed slightly decreased factor X levels and normal factor X antigen and were considered heterozygous for the abnormality. The toponym factor X Padua is proposed to indicate this peculiar abnormality.

The importance of simple coagulation tests (fibrinogen assays and thrombin coagulase clotting time) in the diagnosis of liver cancer.

Dysfibrinogenemia in 36 patients with primary hepatocarcinoma and in 25 patients with cirrhosis of the liver was studied by means of reptilase time, thrombin coagulase time, fibrin polymerization and fibrinogen assays. Both groups of patients were similar in age, sex and incidence of HBs Ag. No electrolyte or fluid imbalances were present. Prolonged reptilase time and prolonged polymerization time were found in both groups; however, thrombin coagulase time was prolonged in 80% of the hepatocarcinoma group, but was normal in almost all patients with cirrhosis (p less than 0.001). In the hepatocarcinoma group, a difference of more than 100 mg per 100 ml was present between the immunologic and coagulative methods of fibrinogen determination in 36.1% of the cases, but in the cirrhotic group this difference was not present in any of the patients (p less than 0.01). We also found that by simply measuring fibrinogen levels by the Mancini method, we could distinguish hepatocarcinoma from cirrhosis in most cases.

Antithrombin III Trento. A 'new' congenital AT III abnormality with a peculiar crossed-immunoelectrophoretic pattern in the absence of heparin.

A family with a new congenital abnormality of antithrombin III (AT III) is presented. 5 members, all females, were affected. The proposita has had several thrombotic manifestations. The other patients, so far, are asymptomatic. Antithrombin activities were all decreased regardless of the method used (chromogenic or clotting) and regardless of the presence or absence of heparin in the assay system. AT III antigen, on the contrary, was normal in all patients regardless of the method used (electroimmunoassay, radial immunodiffusion or Laser nephelometer). The crossed immunoelectrophoresis without heparin showed in plasma the presence of an abnormal peak which was more anodal than the normal counterpart. The same pattern was seen in serum. In the heparin-modified cross-immunoelectrophoresis a normal pattern was seen in plasma and an abnormal one in serum. In the latter the anodal peak was in fact larger than the normal counterpart. Chromatographic studies using Heparin-Sepharose column failed to show changes in heparin affinity, and indicated that both the normal and the abnormal proposed to describe this abnormality. These studies further emphasize the great heterogeneity of AT III defects. This is the first AT III abnormality to show an abnormal crossed-immunoelectrophoresis in the absence of heparin.

Antithrombin III (AT III) Padua2: a "new" congenital abnormality with defective heparin co-factor activities but no thrombotic disease.

A "new" antithrombin III (AT III) abnormality is described in five members of the same family. None of the affected members showed thrombotic manifestations and no consanguinity was present in the family. The main laboratory features were: normal routine clotting tests, slightly decreased AT III activities in all assays carried out in the presence of heparin. In the absence of heparin, antithrombin III activities were instead within normal limitis. Progressive AT III activity and AT III antigen were also normal. Crossed immunoelectrophoresis in the absence of heparin showed a normal pattern both in plasma and serum. In the presence of heparin, the propositi's plasma showed a major, less anodal, abnormal peak and a smaller normal peak. Three peaks were present in the propositi's serum as compared with the two normal ones. This AT III abnormality is different from AT III Padua previously described by us and we propose the toponym of Antithrombin Padua-2 to define this condition.

Antithrombin III Padua: a "new" congenital antithrombin III abnormality with normal or near normal activity, normal antigen, abnormal migration and no thrombotic disease.

A "new" antithrombin III abnormality is described in four members of a family. The proposita is a 38 years old female who showed no thrombotic disease and the following laboratory pattern: normal routine clotting tests, normal or near normal AT III activity (chromogenic substrates S-2238 and Chromozym Th) both in plasma and in serum and in the presence or absence of heparin, slightly decreased antifactor Xa activity (chromogenic substrate S-2222), normal progressive antithrombin, normal AT III antigen but abnormal migration in the agarose-heparin bidimensional system. In the latter test, one major abnormal peak, less anodal than the normal counterpart, and a smaller, apparently normal peak, were seen. In agarose without heparin the pattern was similar to normal both in plasma and in serum. Heparin tolerance to heparin in vivo and in vitro was slightly increased but still within normal limits. The two sons and a paternal aunt showed the same pattern. The hereditary pattern seems therefore autosomal dominant. The abnormality described appears different from AT III Budapest. The toponym of antithrombin III Padua is proposed to define this peculiar abnormality.

Increased factor VIII associated activities in Cushing's syndrome: a probable hypercoagulable state.

Thirteen women and 2 men affected by Cushing's syndrome were investigated. The following parameters were used: plasma and urinary cortisol levels, factor VIII assay (antigen, activity and von Willebrand factor) together with other coagulative assays. Samples were taken before surgery or before medical and/or radiation therapy and every 30-50 days after treatment and continued for 11 months. Cortisol and factor VIII were increased before treatment and decreased slowly after treatment to become normal in 3-4 months. Other clotting tests did not show significant changes. It seems that high plasma cortisol levels may stimulate the production of factor VIII. Patients with Cushing's syndrome often exhibit thromboembolic complications after surgery. It is likely that the clotting abnormalities responsible for such complications is the increased factor VIII activities level.

Demonstration of a double hereditary pattern for congenital afibrinogenemia.

Six patients with congenital afibrinogenemia belonging to four kindreds were studied with regard to hereditary pattern. In two families the hereditary pattern appeared to be autosomal recessive; in the two other pedigrees, on the contrary, the pattern seems autosomal intermediate. In the first type, all family members, excluding the patients, showed normal fibrinogen levels; in the second type, family members could be divided into two groups: normal and heterozygotes. The heterozygotes had fibrinogen levels of 192 +/- 30 mg/dl, definitely lower than that of a normal control population. The average level of the normal relatives was 361 +/- 81.9 mg/dl, practically identical to that of a normal control group unrelated to the homozygotes. In the past these differences were thought to be secondary to variances in fibrinogen assays from one laboratory to the other. It now appears that they are real ones since they can be observed in the same laboratory using the same fibrinogen technique. It must be concluded that congenital afibrinogenemia shows two patterns of hereditary transmission, one autosomal recessive and the other autosomal intermediate.

An immunological study of prothrombin in liver cirrhosis.

Prothrombin was assayed in 24 patients with liver cirrhosis both as activity and as antigen. Prothrombin was found to be decreased by all methods and a good correlation was found among the three methods used. The serum prothrombin assay by means of the electroimmunoassay showed levels comparable to those observed in plasma, as expected. In the bidimensional electrophoresis system, plasma prothrombin appeared decreased but showed a normal mobility. Serum prothrombin in the same system showed the presence of three normal albeit reduced peaks or precipitates.

Hemophilia B with associated factor VII deficiency: a distinct variant of hemophilia B with low factor VII activity and normal factor VII antigen.

Factor VII activity and cross-reacting material was assayed in fresh and deep frozen non-contacted plasma in 43 patients with Hemophilia B belonging to different kindreds. Factor VII activity was found to be slightly decreased (about of 50% normal) in 12 patients, regardless of the thromboplastin used. In an additional patient (hemophilia BM) factor VII was slightly decreased in 1 : 10 diluted plasma but was normal in further diluted plasma. In the remaining 30 patients factor VII activity was normal. No significant variation was found between fresh and deep frozen plasmas. Factor VII antigen or cross-reacting material was normal.

Electroimmunoassay of factor IX in patients with liver damage and vitamin K unresponsive coagulation disorder.

Factor IX antigen and activity were assayed in a group of patients with liver disease and in a group of patients in coumarin therapy. In patients with liver disease there was a similar decrease in activity and antigen. On the other hand factor IX activity is decreased in coumarin treatment with factor IX antigen remaining normal. Factor IX electrophoretic mobility in liver disease is normal.

Studies on factor XIII antigen in congenital factor XIII deficiency. A tentative classification of the disease in two groups.

Immunological and immunofluorescent studies carried out on plasma and platelets of three cases of congenital factor XIII deficiency are reported. Two of these patients were originally thought to have normal factor XIII subunit S and no subunit A. However, repeated assays carried out using different lots of antiserum showed that in reality the patients lacked both subunit S and subunit A. The false positive finding was due to the presence of a anti-factor VIII contaminant in the antiserum originally used. The third patient had a normal subunit S and no subunit A. No factor XIII antigen was found by the indirect immunofluorescent technique in normal, factor XIII deficiency and von Willebrand's disease platelets. On the contrary, by using the non-monospecific antiserum a fluorescent pattern similar to that observed by using an anti-factor VIII antiserum, had been noted. On the basis of the data presented in this paper a tentative classification of factor XIII deficiency in two groups is proposed: Type I is characterized by the lack of both factor XIII subunit S and A. Type II is characterized by a normal subunit S and no subunit A. The need for a re-evaluation of published case of factor XIII deficiency by means of monospecific antisera is indicated.