Kallikrein-kinin system in patients with neoplastic diseases.

The concentrations of kininogens, prokallikrein, fibrinogen, plasma protein, proteinase inhibitor antigens, and the kininase, fibrinolytic and antipapain activities in plasma of 29 patients with various tumours (lung cancers, lymphomas, and others) during chemotherapy were measured. The same studies were performed on blood plasma of female patients operated on because of cervical or endometrial carcinoma (37 subjects) and submitted to subsequent local radiotherapy (before and after radiotherapy). A weak activation of kallikrein-kinin system and significant decrease in kininase and fibrinolytic activities were found in those patients. It may suggest a role of bradykinin in prevention of therapy-induced hypercoagulability in patients with neoplastic diseases.

The effect of local radiotherapy on kallikrein activity in saliva secreted by parotid gland in patients with head and neck cancers.

The samples of parotid gland saliva, collected from control human subjects and those taken from patients with head and neck cancers were submitted to the assay of protein concentration and kininogenase and amidolytic kallikrein activities. No patients with parotid gland tumours were included. The effect of pilocarpine stimulation on these parameters was studied. It was found that the saliva secreted by parotid gland of the investigated patients contains less protein and lower kininogenase activity in comparison to control subjects. Pilocarpine administration resulted in an increase of protein concentration and a decrease of kallikrein activity both in control and investigated subjects. Radiotherapy did not evoke any significant changes in spontaneously secreted saliva. The radiotherapy resulted in a progressive decrease of protein concentration and kallikrein activity in saliva secreted by pilocarpine stimulated glands. The kallikrein activity per mg of protein contained in spontaneously secreted saliva increased significantly during radiotherapy but it distinctly decreased in the saliva of pilocarpine-treated patients. It allows to conclude that the parotid glands do not lose their ability to synthesize and secrete kallikrein during radiotherapy of head and neck cancers.

Plasma kininogen concentration: the low level in cord blood plasma and age dependence in adults.

Antigenic concentration of total kininogen, kinin liberated in vitro, and the antigenic concentration of high molecular weight (HMW) kininogen was measured in 58 different samples of cord blood plasma and in plasma samples from 67 healthy blood donors. Total kininogen and kinin concentration in cord blood plasma was more than twice as low as in pooled plasma of adult persons, and the concentration of HMW-kininogen in cord blood plasma was close to one-third of normal. The concentration of total kininogen and of HMW-kininogen increased with age in adults. All these findings were highly statistically significant.

Striking feature of alpha-fetoprotein (AFP): stimulation of the phagocytic capabilities of human blood platelets.

In concentrations from 2.5 x 10(3) to 480 x 10(3) ng/ml AFP is able to increase the phagocytic index and the phagocytizing percentage of human blood platelets in vitro conditions. This effect is dependent on the dose of AFP and time of incubation.

Alpha fetoprotein inhibits aggregation of human platelets.

The effect of human alpha-fetoprotein (AFP) on platelet aggregation induced by physiological (ADP, PAF, collagen, arachidonic acid) and aphysiological (ionophore A 23,187) activators was investigated. It was found that AFP at the concentrations of 60-750 micrograms/ml, i.e. far below that observed in human blood, inhibits platelet aggregation induced by physiological activators. Platelet aggregation induced by arachidonic acid was two-four times more sensitive to inhibition by AFP than that induced by other physiological agonists, whereas aggregation induced by A 23,187 was not affected by AFP even at concentration of 750 micrograms/ml.

[Plasma kininogenesis and fibrinolysis in the pathogenesis of EPH gestosis]. / Plasmakininogenese und -fibrinolyse in der Pathogenese der EPH-Gestose.

The coexistence of arterial hypertension and disturbances of haemostasis in pregnant women with EPH-gestosis allow to expect a role of fibrinolysis and kinin-forming systems in pathomechanism of this syndrome. For these reasons blood plasma of 34 patients with EPH-gestosis, 23 patients with normal pregnancy and 19 nonpregnant women was investigated. All pregnant women were in third trimester of pregnancy. The following parameters were investigated: kinin-forming system compounds (kininogens and prokininogenases - biological methods), fibrinolytic activity (plasma euglobulin fibrinolysis time), total plasma protein and fibrinogen concentration, protease inhibitors - antithrombin III, C1-esterase inhibitor, alpha 2-antiplasmin, alpha 1-protease inhibitor and alpha 2-macroglobulin (by electroimmunodiffusion). Furthermore hematocrit was measured. In pregnant women with EPH-gestosis significant increase of high molecular weight kininogen concentration was found (p less than 0.02), decreased fibrinolytic activity (p less than 0.01) and (except alpha 2-antiplasmin) decreased concentration of protease inhibitors (p less than 0.005 - p less than 0.01) were observed. Further statistical analysis demonstrated positive correlation between the concentrations of kininogens and prokallikrein-prokininogenases and between low molecular weight kininogen and plasma euglobulin fibrinolysis time. On the other hand negative correlation between concentrations of those proenzymes and severity of gestosis was observed. The above described phenomena indicate on significance of disturbances of proteolytic enzyme activation in pathogenesis of EPH-gestosis.

Culture conditions modify the effects exerted by human fetal alpha-fetoprotein (AFP) on some lymphocyte functions in vitro.

Incorporation of (methyl-3H)-thymidine to DNA of allogeneically and phytohaemagglutinin (PHA)-stimulated lymphocytes cultured in the presence of various concentrations of AFP from human blood cord serum was investigated. The cells were kept in Parker medium supplemented with autologous plasma, calf serum or human AB serum. AFP in concentrations of 5-75 micrograms per one millilitre of culture caused various degrees of inhibiton of responding lymphocyte proliferation. These effects were dependent on the composition of the culture medium, type of lymphocyte stimulation and their individual properties.

Culture medium components modify the effects exerted by alpha-fetoprotein (AFP) on lymphocyte response.

Incorporation of [methyl-3H]thymidine to DNA of lymphocytes cultured in the presence of various concentrations of alpha-fetoprotein (AFP) from human cord blood serum was investigated. It was found that AFP by itself does not exert mitogenic effects. In phytohemagglutinin (PHA) stimulated cultures it may inhibit DNA biosynthesis or exert augmentative effects on this process dependent on the composition of the culture medium.

Human alpha-fetoprotein/AFP/ I. Isolation of homogenous AFP from cord blood serum.

The AFP from human cord blood was isolated by means of affinity chromatography with the use of antibodies as ligands and by gel filtration. The preliminary purification was achieved by affinity chromatography on CNBr-Sepharose 4B coupled with anti AFP-antibody. Further purification was obtained by the use of immunoadsorbent with anti-human serum protein antibodies. Final purification was achieved by gel filtration on Sephadex G-200. Homogeneity of the purified AFP was demonstrated by means of gel filtration, polyacrylamide gel electrophoresis, isoelectric focusing and immunoelectrophoresis.

Inhibition of blastic transformation and immunoglobulin release in lymphocytes cultured in the presence of alpha-fetoprotein (AFP).

Blastic transformation of phytohemagglutinin-stimulated lymphocytes in cultures containing various concentrations of homogenous alpha-fetoprotein (AFP) was studied. It was found that AFP in concentrations of 4-12 microgram/ml inhibits both blastic transformation and immunoglobulin release (IgA, IgG) in lymphocyte cultures. It is suggested that AFP plays an important role in immunoregulation during the fetal life.

Studies of the plasma kinin-forming system in cirrhosis of the liver.

Kininogen level, that of active kinins and kininase activity in the plasma of patients suffering from cirrhosis of the liver and of healthy people were studied. The kininogen content was determined by different available methods i.e. the trypsin and acetone techniques and by means of the plasma and glandular kallikrein preparation. An increase in kininase activity and a lowered kininogen level as determined by all the methods were found in the sick persons. The maximal decrease in the kininogen level determined by means of the plasma kallikreins is substantiated in connection with the increased fibrinolytic activity of the plasma of the patients with hepatic cirrhosis.