Serum antithrombin in coronary-artery disease.

Serum antithrombin activity (AA) was correlated with coronary angiographic findings in 69 patients with documented angina. There was excellent correlation between normal values and normal coronary circulation or only one-vessel stenosis in 30 of 35 patients (86%). When AA was above 90%, 90% of patients (20 of 22) had normal circulation or one-vessel occlusion. In 24 patients AA values were significantly decreased. Coronary angiography in this group revealed three with normal circulation or only one-vessel involvement (10%); 21 of 24 had two or three vessels occluded (90%). The correlation between severe CAD and low AA is probably coincidental to a "triggered" or "turned-on" clotting system. The most practical clinical contribution of AA estimation relates to this capacity to identify angina patients in whom clot-preventive measures (aspirin; dipyridamole; anticoagulants) might prove beneficial.

Blood elastolytic activity in patients with aortic aneurysm.

The question of why obstruction of the aorta (Leriche's syndrome) develops in some patients with severe atherosclerosis of the abdominal aorta while an abdominal aortic aneurysm occurs in others was examined. Significant differences in age, height, weight, mortality, and subsequent operative treatment were found between 335 patients with Leriche's syndrome and 103 patients with abdominal aortic aneurysm. Almost all in both groups smoked and demonstrated leukocytosis. In smokers with aneurysm, circulating serum elastolytic activity and leukocytic granular elastolytic activity were significantly increased, whereas serum antiproteolytic capacity was reduced. These results indicate that the development of abdominal aortic aneurysm in patients who smoke correlates with an abnormal homeostasis between proteolytic and antiproteolytic activity.

Relationship between preoperative status of the fibrinolytic system and occurrence of deep vein thrombosis after major abdominal surgery.

This study comprised 50 patients subjected to major abdominal surgery, of which 13 developed deep vein thrombosis (DVT) according to the 125I-fibrinogen test. Plasma was sampled preoperatively, for the specific analysis of tissue plasminogen activator (t-PA) before and during venous occlusion. The recently described fast t-PA inhibitor and plasmin alpha 2-antiplasmin complex (PAP) were also measured. The result of the laboratory analyses were correlated to the development of DVT. From the data obtained it is concluded that the evaluation of t-PA release during venous occlusion is a poor predictive factor for the occurrence of DVT after major abdominal surgery. The level of the t-PA inhibitor appears to be raised in these patients, but the values obtained in this material were not related to the development of postoperative DVT. Patients with elevated PAP levels, as shown previously, have a lesser tendency to develop postoperative DVT.

Studies of protease inhibitors in the sera of patients with amyotrophic lateral sclerosis.

Serum levels of 4 protease inhibitors, alpha-1-antitrypsin, C1-inactivator, alpha-2-macroglobulin and antithrombin-III were measured in 11 patients with amyotrophic lateral sclerosis (6 males and 5 females) and a control group without neurologic disease. Our results indicated no significant differences in the level of serum alpha-2-macroglobulin between the 2 groups. We found slight but significantly lower levels of serum antithrombin-III in ALS. The possibility of the participation of proteases or protease inhibitors in the pathogenesis of ALS is discussed.

Detection of lysozyme and alpha 2-macroglobulin--lysozyme complexes by immunoblotting.

An immunoblotting technique was developed to detect human lysozyme and lysozyme complexes in body fluids. The unoccupied binding capacity of proteins was demonstrated by addition of surplus lysozyme. The sensitivity of immunoblotting to the free enzyme in human albumin solution was less than 5 ng. In serum and pleural fluid, part of exogenous lysozyme was bound to alpha 2-macroglobulin (alpha 2-M). At high concentrations of lysozyme in leukemic sera, part of the enzyme formed an endogenous alpha 2-M complex. On the other hand, the formation of alpha 2-M complexes with exogenous lysozyme was especially striking in sera from nephrotic patients with elevated alpha 2-M. The findings corroborate with previous reports on lysozyme binding to purified alpha 2-M in vitro and suggest that the binding is concentration-dependent with respect to both reaction partners. In vivo the mechanism may provide a pathway for extrarenal lysozyme catabolism medicated by reticuloendothelial cells. No other binding proteins were seen in the present study: lysozyme did not bind to serum immunoglobulins in 35 samples with an immunoglobulin paraprotein, three samples with polyclonally elevated gamma-globulins, 20 other patient sera and 10 normal sera. Neither did lysozyme bind to urinary proteins in five samples from patients with myeloic leukemias nor in 10 samples from myeloma patients with urinary excretion of a monoclonal immunoglobulin light chain.

The antiproteolytic compound alpha 2-macroglobulin in human skin.

Monoclonal antibodies recognizing the antiproteolytic compound alpha 2-macroglobulin (MG) were used for immunohistological studies on normal human skin. MG-specific immunoreactivity was found to be localized to the papillary dermis and to be concentrated in the region of the epidermodermal junction. In view of these findings and the possible involvement of proteolytic enzymes and their inhibitors in blister formation, we asked whether MG occurs in the fluid of experimentally induced blisters. MG was identified (by western-blotting) and quantified (by a monoclonal antibody based enzyme immunoassay) in the fluid of experimentally induced suction blisters. Taken together, MG is present in such blister fluid in concentrations 6 times lower than in serum, but still in an antiproteolytic range. These findings allow suggestion of a possible role for the antiproteolytic compound MG in blister formation.

Tuberous sclerosis: the incidence of sporadic cases versus familial cases.

In 48 families in which tuberous sclerosis occurred, extensive examination presented almost the same incidence of sporadic cases as reported in previous studies. Although inspection of the skin and cranial computed tomography seem to be the most sensitive diagnostic tests available, negative results with these methods do not exclude the diagnosis. Estimation of alpha 2-macroglobulin serum level does not mean an extension of the diagnostic arsenal.

Serum alpha-1 antichymotrypsin is a possible growth inhibitor of Plasmodium falciparum.

Serum protease inhibitors were determined in paired sera from 7 patients with cerebral malaria and 2 patients with acute malaria showing high and low growth inhibition activity in the initial and follow-up sera respectively. Alpha-1 antichymotrypsin and alpha-1 antitrypsin but not alpha-2 macroglobulin showed direct correlation with the growth inhibition activity. When alpha-1 antitrypsin was deliberately added to the malarial culture no growth inhibition occurred indicating that the alpha-1 antichymotrypsin was the most likely factor responsible for inhibition of growth of malarial parasites in vitro.

[Activation of the kallikrein-kinin system of the blood in hypertension and atherosclerosis with transient cerebral circulatory disorders]. / Aktivnost' kallikrein-kininovoi sistemy krovi u bol'nykh gipertonicheskoi bolezn'iu i aterosklerozom s prekhodiashchimi narusheniiami mozgovogo krovoobrashcheniia.

The authors studied the blood kinin system in patients with transient impairments of the cerebral circulation (TICC) both in the presence of hypertonic crises (30 patients) and cerebral vascular atherosclerosis (30 patients) during a vascular episode on the 10th-15th day of treatment. During TICC, the patients showed a marked activation of the blood kinin system which was especially pronounced in hypertonic rises. The underlying disease had a significant impact on the time-course of kinin system activity. In patients with TICC associated with hypertonic crises, hypotensive treatment decreased kinin activity to a considerable degree whereas in cases of cerebral vascular atherosclerosis the treatment exerted no noticeable effect on the activity of kinins. The involvement of kinins in the processes of the compensation and pathogenesis in TICC is discussed. It is recommended that TICC associated with arterial hypertension be treated by sodium salicylate, whereas in TICC associated with atherosclerosis the administration of antikinin drugs is indicated (parmidin, prodectin).

[Status of the kallikrein-kinin system of the blood in young and middle-aged patients with ischemic heart disease]. / O sostoianii kallikrein-kininovoi sistemy krovi u bol'nykh ishemicheskoi bolezn'iu serdtsa molodogo i srednego vozrasta.

Increase in BAEE-esterase and kallikrein activities, simultaneously with decrease in alpha 2-macroglobulin and prekallikrein activities were found in blood plasma of young and middle age patients with heart ischemic disease. The maximal alterations were observed in young patients as well as in patients, subjected to heart infarction less than a year before the examination. Maximal activity of kallikrein was accompanied by deterioration of fibrinolytic activity, by hypertriglyceridemia and by increase in adrenaline excretion. Dietetics normalized partially the state of kallikrein-kinin system, especially, in patients ration of which contained soybean products.

Immunofluorescent evaluation of platelet alpha-1-antitrypsin and alpha-2-macroglobulin.

Rabbit raised anti-alpha-1-antitrypsin or anti alpha-2-macroglobulin antisera at dilution of less than 1:80 yielded non-specific staining on human platelets by indirect immunofluorescent technique. A similar pattern was in fact obtained by using normal rabbit sera at the same dilution and was due to the presence of smooth muscle autoantibodies. This indicates that human platelets do not contain significant quantities of these antigens. In agreement with the above, only microamounts of alpha-1-antitrypsin and alpha-2-macroglobulin were found to be present in human platelets by means of the electroimmunoassay.

The elimination of trypsin--alpha-macroglobulin complexes from the blood circulation in pigs during acute pancreatitis and after massive intravenous trypsin infusion.

The elimination from plasma of intravenously injected 125I-labelled trypsin-alpha-macroglobulin complexes was studied in both healthy and diseased, anaesthetized pigs. The t1/2 for the 125I-labelled complexes was 5-10 min in healthy pigs as well as in two groups of pancreatitis pigs. Pre-loading of the eliminating capacity by massive intravenous trypsin infusion, raised t1/2 to 80-100 min. Thus, the eliminating capacity seems to be saturable. Elimination of the complexes from the blood circulation occurred within 4-5 h. Even massive intravenous amounts of trypsin-alpha-macroglobulin complexes gave no demonstrable complement or kinin activation. Neither were other deleterious effects seen during the experiment or during 7 days of follow-up. The results indicate that treatment of acute pancreatitis in humans with alpha 2-macroglobulin might be possible, without any danger arising from the protease-alpha 2-macroglobulin complexes formed.

Induction of the "pregnancy zone" protein by conjugated oestrogens.

Treatment with conjugated oestrogens was found to increase significantly the serum concentration of the pregnancy zone protein (PZ) in a group of postmenopausal women. The basic level of PZ before treatment was comparatively high in this group of women and might be related to age and previous pregnancies. Naturally occurring conjugated oestrogens have a similar effect on the induction of PZ as synthetic oestrogens.

PIP: The concentration of pregnancy zone (PZ) protein was measured by radioimmunoassay in postmenopausal women before and during treatment with conjugated estrogens (1.25 mg Promarit, 6 days/week for 2 months). The mean serum concentration of PZ in 34 women before treatment was 5.3 plus or minus .6 mg/100 ml. After treatment the mean serum concentration for 31 women was 13.9 plus or minus 1.7 mg/100 ml. The increase was significant (p less than .01). The level of PZ in these women was higher than usual and this is attributed to their age and to their previous pregnancies. Naturally occurring estrogens and synthetic estrogens have a similar effect on the induction of PZ.

[alpha 2-Macroglobulin in serum and plasma: reference values with carbobenzoxy-valyl-glycyl-arginine-p-nitroanilide, a chromogenic substrate]. / alpha 2-Makroglobulin in Serum und Plasma: Referenzwerte mit Carbobenzoxy-valyl-glycyl-arginin-p-nitroanilid, einem chromogenen Substrat.

alpha 2-Macroglobulin can be determined using the amidolytic activity of the alpha 2-macroglobulin-trypsin-complex. Reference values are reported using a commercially available kit for continuous assay at 25 degrees C. Reference ranges obtained in a population of 235 children and adults aged between 1 day and 61 years were found to be age-dependent. Reference ranges established for the different age groups of children were grouped together in order to obtain useful guide values. The following values were obtained for serum: 1 to 10 days: 5-11 kU/l 11 days to 18 years: 8-16 kU/l The reference values (5th to 95th percentile) determined for juveniles and adults were as follows: 19 to 25 years: 4.1-12.2 kU/l (serum); 4.0-11.2 kU/l (citrated plasma) greater than or equal to 25 to 61 years: 4.1-8.1 kU/l (serum); 3.7-7.4 kU/l (citrated plasma) No variation according to sex was observed, nor was any difference noted between preprandial and postprandial catalytic concentrations, or between values in smokers and non-smokers or between values in the group of women taking hormonal contraceptives and the group not taking such drugs. If the dilution of the blood sample with citrate solution is taken into account when calculating the catalytic concentrations in citrated plasma, the values in plasma are higher than those in serum (median 11%). Measurement of the alpha 2-macroglobulin-trypsin complex in plasma is assumed to give a more realistic reflection of the in vivo activity than measurement in serum. Comparison with two immunological methods (radial immunodiffusion and kinetic nephelometry) revealed that there is a linear relationship between the measurement of the catalytic concentration of the alpha 2-macroglobulin-trypsin complex and the immunological concentration of alpha 2-macroglobulin. Therefore mutual conversion is possible for the described concentration range.

alpha 2-Antiplasmin and alpha 2-macroglobulin--the main inhibitors of fibrinolysis--during the menstrual cycle, pregnancy, delivery, and treatment with oral contraceptives.

alpha 2-Antiplasmin and alpha 2-macroglobulin have been studied during the menstrual cycle, pregnancy and parturition in healthy women, and during use of various types of contraception in both healthy and diabetic women, and compared with a reference group of healthy men and women. alpha 2-Antiplasmin showed a slight sex difference, with higher values in women. The luteal phase of the menstrual cycle showed slightly higher values than the other phases. alpha 2-Antiplasmin increased during pregnancy, decreased (probably due to consumption) during labor and increased again in the puerperium. Treatment with neither combined contraceptive pills nor low dose progestogen pills gave any changes in alpha 2-antiplasmin. alpha 2-Macroglobulin showed low values during menstruation. The increase during pregnancy and treatment with combined contraceptive pills is in accordance with earlier findings. It is concluded that synthesis and metabolism of alpha 2-antiplasmin are under hormonal influence. The role of alpha 2-antiplasmin in the decreased fibrinolysis in pregnancy is discussed.

Serum zinc in athletes in training.

Serum zinc was determined using atomic absorption spectrophotometry in 160 training athletes (57 females) in the morning at rest. In 23.3% of the male and 43% of the female athletes, serum Zn was lower than the limit accepted for the normal range (75 microgram/dl or 11.5 mumol/l). The average serum Zn (96.7 +/- 12.6 microgram/dl) and the range of the values found in a control group of 15 young adult males did not differ from the accepted values in the literature. In 22 randomly selected male athletes, serum Zn fractions were determined using polyethylene glycol precipitation at pH 7.1; 22.2% of total Zn was bóund to alpha2-macroglobulin, the ratio being very close to 1 atom of metal per molecule of globulin. The possible reasons that may influence the serum Zn level in athletes are discussed with regard to the present knowledge in both exercise physiology and metabolism of zinc in man.

[Chromatographic separation of activated proteases from human plasma]. / Zur chromatographischen Trennung aktivierter Proteasen aus menschlichem Plasma.

After separation of aceton and dextran sulfate activated human plasma by column chromatography on DEAE-cellulose three esterolytically and amidolytically active fractions, respectively, were obtained, which were assigned to the following species: plasma kallikrein (PK), PK.alpha-macroglobulin.HMW-Kininogen. Their percentage in the whole activity is variable. The proportion of free PK is low (0.11). For characterization of the products we studied inhibition by different polyvalent inhibitors. The Michaelis constant (Km) with p-toluene-sulfonyl-L-arginine methyl ester (TAME) were determined. For simulation of in vivo conditions dextran sulfate activated plasma was inactivated at 37 degrees C. The residual activity and the spontaneous activity in plasma from patients with shock are produced by different active protease inhibitor complexes.

Open quaternary structure of the hagfish proteinase inhibitor with similar properties to human alpha-2-macroglobulin.

A homologous protein to human plasma alpha-2-macroglobulin (alpha-2-M) was purified from the blood plasma of hagfish (Eptatretus buergeri) and its structure and function were studied. The hagfish protein inhibited several proteinases and its inhibitory activity was blocked with methylamine as in the case of human alpha-2-M. The molecular weight and sedimentation coefficient of the hagfish inhibitor were 390,000 +/- 20,000 and 11.0 S, respectively, as determined by sedimentation studies. The frictional ratio calculated from these parameters was 1.75. The Stokes radius estimated from HPLC gel chromatography was 8.8-8.9 nm, which was similar to that of human alpha-2-M despite the fact that the hagfish inhibitor was only one-half as large as human alpha-2-M in molecular weight. The hagfish inhibitor was expected to be more asymmetric and/or more hydrated than the human inhibitor. The electron micrographs of the negatively stained hagfish inhibitor showed that it had an open, rectangular quaternary structure of 15 +/- 1.5 X 19 +/- 2 nm in which two semiglobular units were located at the two shorter sides with a gap of 8 +/- 1 nm in width. Each semiglobular unit had an approximate width of 5 +/- 0.5 nm. The thickness of the unit was estimated to be 3 to 3.5 nm from the result of fixed-angle shadowing experiments. Although the two semiglobular units must be connected by some structure, very little material could be seen between them. Such an open quaternary structure may explain the high frictional ratio and large Stokes radius of this protein. The structural change of the inhibitor after reaction with proteinases or methylamine could be detected by electron microscopy and gel chromatography.

Plasma factor XIII and some other haemostasis parameters in patients with diabetic angiopathy.

Since intravascular and endoparietal fibrin deposition is thought to be involved in the development of atherosclerosis, we measured factor XIII activity and its subunit 'a' and 'b' concentrations against a background of other haemostasis parameters in diabetics with angiopathy and in 2 control groups (healthy subjects and diabetics without vascular complications). Diabetics with angiopathy revealed a significant increase of factor XIII activity as well as its subunit concentrations. They also had significantly elevated anti-thrombin III, alpha 2 macroglobulin, alpha 1 antitrypsin, C1 inhibitor, fibrinogen, FDP concentrations and prolongation of euglobulin lysis time. The highest factor XIII levels were found in diabetics with renal failure. We suppose that increased factor XIII level and other observed changes of haemostasis in patients with diabetic angiopathy might promote intravascular and endoparietal fibrin deposition and contribute to the development of atherosclerotic complications of diabetes.