Differentiation of mitotic melanoma cells from G2 cells and their isolation by use of 5-bromo-2'-deoxyuridine and propidium iodide

This report describes a method by which mitotic cells were isolated from nonsynchronized Cloudman melanoma cells that had been pulse labeled with 5-bromo-2'-deoxyuridine (BrdUrd) and double-stained with a fluoresceinated monoclonal antibody to BrdUrd and with propidium iodide (PI). In initial experiments, melanoma cells were first pulse labeled with BrdUrd, treated with prostaglandin E1 (PGE1 10 micrograms/m1) or vehicle (0.1% ethanol) for up to 24 hours, then stained with anti-BrdUrd and PI. PGE1-treated cells monitored at 3-hour intervals were observed to migrate from S phase to G2 phase, then, enigmatically, back into the late S phase region of the distribution. In other experiments, cells treated with PGE1 were pulse labeled with BrdUrd at the end of the treatment period and harvested. In these experiments, there was a small, discrete subpopulation of cells within the late S phase region of the DNA distribution that was negative for anti-BrdUrd. This subpopulation of cells was sorted and examined by light microscopy. We observed that 95% of these BrdUrd-negative "S phase" cells were mitotic cells. Since mitotic cells and G2 cells have equivalent amounts of DNA, the reduced red fluorescence exhibited by these cells may be due to a greater sensitivity to denaturation, which has been described for DNA of mitotic cells, and would account for the phenomenon of cells appearing to move "backwards" in the cell cycle. This report indicates that although the BrdUrd/PI method can further define the cell cycle into four compartments, it can also lead to over-estimation of S phase cells in kinetic studies because of contaminating mitotic cells.     

Effect of external gamma radiation on shortening the life expectancy and rates of the formation of absorbed doses from incorporated 239Pu in dogs

External gamma irradiation of dogs with doses of 103.2 and 51.6 mC/kg combined with the effect of inhaled plutonium-239 accelerates the formation of absorbed doses in secondary organs of the radionuclide deposition by 41.7 and 2.4 times, respectively, whereas the dose of 25.8 mC/kg is ineffective. As estimated by the rate of 239Pu accumulation and by the life span shortening, the minimum effective and the maximum ineffective doses are 104.8 and 80.5 cGy and 89.2 and 79.2 cGy, respectively.     

The isolation and immunological properties of two arginase forms from human erythrocytes

1. Two forms of arginase were isolated from human erythrocytes; the main form adsorbed on CM-cellulose and the second form, occurring in much smaller amount, adsorbed on DEAE-cellulose. 2. The molecular weight of either arginase was 120,000 +/- 5000. 3. The erythrocyte arginases are similar in immunological properties to arginase A4 from human kidney and A2 from human liver, respectively. 4. Despite the literature data stating that human erythrocyte arginase and human liver arginase are identical, it was found that the main forms of arginase of these tissues A4 from erythrocytes and A5 from liver differ in immunological properties.     

Histochemistry of proteases in ependyma,choroid plexus and leptomeninges

Summary Aminopeptidase M (APM), aminopeptidase A (APA), dipeptidyl peptidase IV (DPP IV) and -glutamyl transferase (GGT) were demonstrated histochemically in cryostat sections of the rat brain to show the reaction pattern of ependyma, choroid plexus and leptomeninges. GGT was only demonstrable in the cell membranes of ependymal cells and in the leptomeninges; however, APA, APM and DAP IV showed a variable degree of activity in the capillary endothelium of the choroid plexus as well as in the leptomeninges. On the basis of these results, it is postulated that peptides in the cerebrospinal fluid can be cleaved extraventricularly by the enzymes demonstrated in the leptomeninges.Dedicated to Professor Dr. T.H. Schiebler on the occasion of his 65th birthday     

Effect of methylguanidine and guanidine succinic acid on methemoglobin reductase activity in human red cells]

Activity of methemoglobin reductase was studied in human red cells treated with methylguanidine and guanidinosuccinic acid in concentrations similar to those in plasma of patients with chronic renal failure. Enzyme activity was measured with Richterich technique following an incubation at 37 degrees C for three hours. Results have shown that methylguanidine in concentration of 5.4 x 10(-5) mol/l decreases activity of methemoglobin reductase in human red cells on average by 13.9%. Higher concentrations potentiate this effect. Similar changes in methemoglobin reductase activity were noted after introduction of guanidine-succinic acid into the mixture. This agent in concentration 5.6 x 10(-5) mol/l inhibited activity of the tested enzyme by 34.2% on average. Combined methylguanidine in concentration of 5.4 x 10(-5) mol/l and guanidine-succinic acid in concentration of 2.8 x 10(-5) mol/l inhibited methemoglobin reductase activity by 33.0% on average. It may be suggested, that methylguanidine and guanidine-succinic acid being low molecular uremic toxins may significantly decrease methemoglobin reductase activity in red cells of patients with chronic renal failure.     

The effect of intracoronary injections of x-ray contrast media on cardiac rhythm and conductivity during different variations of myocardial blood supply]

A study of the correlation between electrophysiologic effects of intracoronary injections of radiopaque agents and anatomical features of blood supply of cardiac conduction was carried out in 60 patients with intact coronary arteries. Coronary angiography was performed in all the patients. Disorders in cardiac conduction and repolarization in the myocardium were observed in intracoronary injection of radiopaque agents, which was accompanied by the development of S-A bradycardia. A bradyarrhythmic reaction type depends on prevalence of the left or right coronary artery in the atrial blood supply and cardiac conduction. The bradyarrhythmic effect was more pronounced in injection of a radiopaque agent in the right artery than in the left one.