Biomedical subjects
Publications and source records attributed to M Szmitkowski.
The paper presents the molecular and kinetics aspects of alcohol dehydrogenase polymorphism. The role of this enzyme in ethanol metabolism are too discussed.
The morphological and biochemical changes of the liver after endotoxin intake were analyzed in rats receiving 20% ethanol during 60 days. Besides morphological changes, concentration of serotonin and histamine in liver homogenates, the activity of asparagine and alanine aminotransferases (AspAT, ALAT), gamma-glutamyltranspeptidase (GGTP) and alcohol dehydrogenase (ADH) in blood serum were determined, too. The most extensive morphologic changes of the liver were seen in group of animals intoxicated with 20% ethanol during 60 days and single dose of endotoxin E. coli 0127:B8 intraperitoneally. These changes included necrosis most hepatocytes, focal steatosis of liver parenchyma, considerable hyperemia and parenchymatous degeneration of the liver cells. The cells lining liver sinuses showed considerable swelling as well as necrotic changes. Figures of cell division and haemorrhagic focuses were seen, too. The clusters of mononuclear cells, surrounding necrotically changed hepatocytes were seen in the central part of the liver lobule. Among the inflammatory mediators estimated in liver homogenate only serotonin reached a high level in the group of experimental animals receiving only endotoxin. Increased activity of aminotransferases AspAt and ALAT were associated with these morphologic and biochemical changes in liver tissue observed in animals receiving ethanol and endotoxin.
The activity of the colony stimulating factor (SCF) was measured in kidney subcellular fractions in mice. The highest activity was noted in microsomes. From other fractions, the cytosol had large amounts of CSF. On the basis of literature data, and the findings presented we suggest that the kidney is at least one of the organs of CSF biosynthesis.
Substances with CSF activity were studied in human and murine sera. Gel filtration of the sera on Sephadex G-200 showed that fractions with molecular weight of 14,000 and 6000 D were active in the CSF test. When mice were stimulated with endotoxin, CSF activity of both fractions was twice as high as in control animals, and CSF activity was found also in a protein peak with the molecular weight of 300,000 D. The fractions with CSF activity were sensitive to pepsin.
We found one peak of a new molecular form of CSA after chromatography on Sephadex G-200 in the serum of mice treated with endotoxin. Total activity and two peaks of CSA were much higher than controls.
Human sera after column chromatography on Sephadex G-200 were tested for Granulopoietic Activity (GA) in vivo and Colony Stimulating Factor (CSF) in vitro. We have found that GA after chromatography is distributed in three peaks but CSF only in one peak with high activity, and evident activity in volume 135 ml. This activities are in good agreement with two peaks of GA. We assume that GA and CSF after this chromatographic procedure is free from lipoproteins inhibitors. The obtained results are briefly discussed.
Factor VIII has been isolated and purified from human granulocytes using chromatography on Sephadex G-200 AND DEAE-Sephadex A-50. The final preparation was purified 60-fold. Some properties of this partly purified factor VIII were compared with the human plasma AHF preparation. Optimum pH activity and the highest activity after incubation at different pH values was found in buffer at pH 7.0. The preparation was thermolabile and had a molecular weight of about 214,000. Absorption curves indicated that it is a protein exhibiting four fractions in polyacrylamide gel disk electrophoresis. AHF preparations from plasma and granulocytes were almost identical in the characteristics tested.
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The effect of cytotoxic drugs was tested on the hematopoietic system by assay of the ability of granulocyte-macrophage colony forming cell (GM-CFC) in mice to create GM colonies. The in vivo ability of bone marrow progenitor cells to granulocyte-macrophage colony formation, was tested after long-term peritoneal cytotoxic drug administration. The direct effect of these agents on cells in vitro culture was evaluated also. It was found that cytotoxic drugs inhibit the GM colony formation. The degree of damage to the bone marrow progenitor cells by assaying in vivo colony formation inhibition, depends on the drug dosage and length of therapy (after 25-30 days of treatment the colony growth was below 50%). The in vitro inhibition of granulocyte-macrophage colony formation depends on the concentration of drug (10(-7)-10(-5) M is critical for GM colony growth). The results suggest the possibility of the GM-CFC growth testing as an indicator of the progress or side effects of cytotoxic therapy.
The effect of glucocorticoids was tested on the hematopoietic system by assay of the ability of granulocyte-macrophage colony forming cells (GM-CFC) in mice to create GM colonies. The in vivo ability of bone marrow progenitor cells to granulocyte-macrophage colony formation was tested after long-term peritoneal glucocorticoids administration. The direct effect of these agents on cells in vitro culture was evaluated also. It was found that glucocorticosteroids inhibit the GM colony formation. The degree of damage to the bone marrow progenitor cells assaying by in vivo colony formation inhibition depends on the drug dosage and the length of therapy. The in vitro inhibition of granulocyte-macrophage colony formation depends on the concentration of the drug. The results suggest the possibility of the GM-CFC growth testing as an indicator of the side effects of prolonged corticoid therapy.
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Activity of CSF was measured in human serum after chromatography on Sephadex G-200. Agar culture of bone marrow cells was done in Petri dishes with agar in Eagle'a medium containing fetal calf serum and antibiotics. This agar-medium containing mouse bone marrow cells, was allowed to gel and the dishes were incubated at 37 degrees C in a fully humidified atmosphere of 10% CO2 in air. Our data indicate that chromatography procedure allows to get CSF free from its inhibitors. All activity was localized in one peak, in the region of low molecular weight proteins.
The colony stimulating factor and granulopoietic activity were determined in the homogenates of mouse kidney, liver, spleen and bone marrow. The activity of CSF and GA were highest in the kidney. The homogenate of this organ was fractionated then by ultracentrifugation. The distribution of CSF and GA in subcellular fractions and homogenates may suggest that kidney cells are the source of a substance causing, both, colony stimulation and granulopoietic activity.