Populations of thymocytes and peripheral blood leucocytes in leukaemia-bearing mice treated with G-CSF.

The influence of granulocyle-colony stimulating factor (G-CSF) on thymocyte subsets and peripheral blood leucocytes in leukaemia L1210-bearing mice was evaluated. Leukaemia-bearing mice have a markedly reduced L3T4+Lyt2+ thymocyte subpopulation. We observed "correction" of relative values of thymocyte subpopulations induced by treatment of mice with a G-CSF preparation. The suggestion is presented that this could result from liberation of lymphocyte T precursor cells from the bone marrow and increased homing of them into the thymus. The direct or indirect influence of G-CSF on the L3T4+Lyt2+ subpopulation of thymocytes is discussed.

Thymocyte and splenocyte subpopulations in normal and leukemia-bearing mice after adrenalectomy.

Mice with i.p. inoculated leukemia L1210 cells had lower total number of thymocytes, and reduced number of CD4+8+ thymocytes, but increased CD4+8- subpopulation. Adrenalectomy reduced the CD4-8+ subset of thymocytes. Leukemia in adrenalectomized animals induced similar changes in the number of thymocyte subpopulations as observed in normal animals. However, a decrease in CD4+8- thymocytes was noted and the animals had decreased number of CD4-8- cells as well. Splenocytes of CD4+8- subpopulation were more numerous in adrenalectomized animals. Leukemia induced increase in the number of CD4+8- and CD4-8+ splenocytes in both normal and adrenalectomized animals.

[Plasma prekallikrein in chronic liver diseases]. / Prekalikreina osoczowa w przewleklych chorobach watroby.

Plasma prekallikrein (PPK) is a single-chain glycoprotein synthesized in the liver. The aim of our study was to evaluate a plasma prekallikrein as the marker reflecting liver protein synthesis in patients with chronic liver diseases. PPK levels have been measured by own modification of amidolytic micro-assay in 43 patients with chronic liver diseases and 37 healthy volunteers as control group. As compared to control group, PPK level was significantly decreased in patients with chronic active hepatitis and with decompensated liver cirrhosis and significantly increased in patients with liver cirrhosis complicated by hepatocellular carcinoma. There was no difference in plasma prekallikrein between patients with compensated liver cirrhosis and controls. The results suggest that PPK might be a useful index for the assessment of residual functional liver mass in patients with chronic liver diseases.

Synthesis of novel, aminoalkanolic derivatives of 5,7-dihydroxy-2-phenylbenzopyran-4-on as the potential cytostatic agents.

The 7-(2,3-epoxypropoxy)-2-phenyl-5-hydroxy-4H-benzopyran-4-on and its aminoalkanolic derivatives were prepared as the potential cytostatic (antileukemic) compounds. They were tested in vivo on mice leukemia L1210.

[Elevated plasma prekallikrein level in patients with diabetes]. / Podwyzszony poziom prekalikreiny osoczowej u chorych na cukrzyce.

The contact activation of intrinsic pathway in the coagulation system accompanied by plasma kallikrein-induced kinin generation is thought to be involved in the pathogenesis of diabetic retinopathy. Plasma prekallikrein (PPK), a proenzyme of plasma kallikrein, is a single-chain glycoprotein synthesized mainly in the liver. The aim of our study was to evaluate plasma prekallikrein level in diabetic patients and to examine the relationship between PPK and the metabolic control of diabetes and development of retinopathy. In 53 diabetic patients and 33 healthy subjects as controls the following parameters have been assessed: plasma prekallikrein, serum fructosamine, glycated haemoglobin HbA1c, prothrombin time, partial thromboplastin time and antithrombin III (AT III). Compared to the control group, PPK level was significantly higher in diabetics, especially in patients with proliferative retinopathy. The significant positive correlations have been found between PPK and HbA1c in diabetic patients and between PPK and serum fructosamine concentration but only in diabetics without retinopathy. No differences in prothrombin time and AT III have been observed between diabetics and healthy subjects. A suggestion is presented on increase of plasma prekallikrein level in diabetics due to hyperglycaemia-stimulated glycoprotein over-synthesis in the liver, what would confirm the role of kallikrein-kinin system in the pathogenesis of microangiopathy.

Plasma prekallikrein levels in patients with hepatocellular carcinoma and liver cirrhosis: a pilot study.

The concentration of plasma prekallikrein (PK) in five patients with hepatocellular carcinoma (HCC) has been measured and related to levels in 18 patients with liver cirrhosis (LC) and 30 healthy subjects. It was found that the mean PK level was significantly increased in patients with HCC, while patients with LC demonstrated lower concentrations, as compared with healthy subjects. The results indicate that PK might be useful in screening cirrhotic patients for HCC. Longitudinal studies of PK in a larger group of patients at risk of developing HCC are therefore recommended.

Changes of thymocyte subpopulations induced by activities diffusing from leukemia L1210 cells.

Mice with intraperitoneally inoculated leukemia L1210 cells had a lower total number of thymocytes and a markedly reduced percentage of L3T4+ Lyt2+ (CD4+ 8+) thymocytes, but increased subpopulations of L3T4+ Lyt2-, L3T4- Lyt2+, L3T4-Lyt2-. The percentage of Thy 1.2+ thymocytes was unchanged as compared with control animals. A similar effect was observed is suggested that leukemia L1210 cells produce some activities which diffuse through 0.22 microns filters and influence the subpopulations of thymocytes.

[Evaluation of prekallikrein activator (PKA) activity in plasma-derived preparations]. / Ocena aktywnosci aktywatora prekalikreiny (PKA) w preparatach osoczopochodnych.

The study was carried out on 111 series of human immunoglobulin preparations (81 series for intravenous use, 30 series for intramuscular use) produced abroad and in Poland. Kallikrein (KK) and prekallikrein activator (PKA) activities were identified using chromogenic substrate. S-2302 and proteinase inhibitors were tested by double immunodiffusion method. Several biochemical, biological and serological methods were also applied which are valid in the control of intravenous and intramuscular immunoglobulin preparations. It was found that trace amounts of PKA and KK are present in human immunoglobulin preparations for intravenous use such as Bioglobulina and Gamma-Venina (PKA 0.0092-0.0505 mu kat/L, and KK 0,-0.0093 mu kat/L). On the other hand normal human immunoglobulin preparations for intramuscular use had somewhat higher level of KK and PKA and anti-HBs Immunoglobulin preparations (5.50 mu kat/L and 153.91 mu kat/L) and Tetabulina preparations had very high levels of these factors as compared to remaining preparations (4.50 mu kat/L and 316.5 mu kat/L).

Proteinase activity and agglutination of leukemia cells.

The proteinase activity was assayed in the leukemia cells L 1210 and in the ascites fluid with [3H] acetylated haemoglobin as a substrate. The proteinase activity at pH 4.1 increased in cells and in the ascites fluid with age of the tumor. The proteinase activity at pH 7.8 was low, but the enzyme activity in the cell homogenate increased between 5th and 7th day of the tumor growth and it was also present in the ascites fluid. It was observed that the leukemia cells aggregate in vivo and in vitro at pH values of the ascites fluid above pH 7.0. It was suggested, that the aggregation of leukemia cells is due to the tumor cell proteinase activity released to the ascites fluid.

125I-labeled cell surface as a marker in preparation of microsome fraction by gel filtration.

The microsomal fraction was isolated from homogenate of 125I-labeled leukemia L 1210 ascites cells by filtration of postmitochondrial supernatant through a Sepharose 4 B column. It was found that the particles are labeled with iodine and show 5'-nucleotidase activity suggesting the presence of cell membranes in the fraction. The soluble proteins fraction were retarded on the column showed lactate dehydrogenase activity, and low activity of soluble beta-D-glucuronidase.

Radioiodination of L 1210 cells.

The lymphoid leukaemia L 1210 cells of mice were labelled with 125I. The cell homogenates were fractionated and from the microsomal fraction 90 per cent of the radioactive material could be precipitated with perchloric acid, whereas only 4 per cent was precipitated from the soluble fraction. Papain bound with Enzacryl AH released 31 per cent of radioactivity. It was concluded therefrom that the surface proteins of the cells were labelled. Electrophoretic separation of these proteins in polyacrylamide gel with sodium dodecyl sulphate was performed and 6--8 radioactive fractions of surface peptides were found.

Synthesis of glucuronoglucans in chicken cartilage cultured in synthetic medium.

Cartilaginous femur and tibia rudiments from 10-day-old chick embryos were grown in vitro for 4 days in Parker's solution without protein added, and subsequently fixed and extracted successively with 0-2 N HClO4 at 4 degrees C (fraction A), 5 per cent trichloracetic acid (TCA) at 4 degrees C (fraction B), and 5 per cent TCA at 90 degrees C (fraction C). The residue after extraction was dissolved in 1 N NaOH at room temperature (fraction D). Fraction C containing most of hexuronic acids and aminosugars of the cartilage was used to study the quantitative changes of glucuronoglucans throughout the culture period. The amount of hexuronic acids and aminosugars was increased after 24, 48, 72 and 96 hours of culture. After 96 hours the level of hexuronic acids was twice that found prior to establishing the culture. The increment was statistically significant.