The dynamics of the endothelial cell population was investigated in the rat brain after local irradiation with different doses of X rays. A fluorescent-histochemical technique was used for the visualization of the cells. A decrease in endothelial cell number was observed within 1 day of irradiation with doses of 5-200 Gy. At this time the endothelial cell number had decreased by up to 15% compared with the pre-treatment values. This early dose-independent loss in cell number was maintained for up to 1 month after irradiation. This was then followed by a slow dose-independent decrease in cell density up to 6 months after exposure. Subsequently the depletion of the endothelial cell population exposed to 40 and 60 Gy continued. After a dose of 25 Gy an abortive recovery of cell numbers occurred followed by an abrupt depletion of the endothelial cell population. The possible mechanisms of such changes are discussed.

The present study demonstrates that DNA fragmentation, nuclear pycnosis and trypan blue staining of irradiated thymocytes is prevented by inhibition of the lipoxygenase pathway of arachidonic acid metabolism and is not affected by cyclooxygenase inhibition. Exposed to irradiation [3H]arachidonic acid-labeled thymocytes release radioactive products to the external medium. The process is blocked by the lipoxygenase inhibitor, nordihydroguaiaretic acid. Thus, it can be concluded that irradiation activates arachidonic acid metabolism and that lipoxygenase metabolites play an important role in thymocyte apoptosis.

In vitro permittivity measurements of excised human liver, spleen, kidney and cardiac muscle at frequencies from 10 kHz to 100 MHz are described. An end-of-the-line capacitive sensor and a computer-controlled network analyser HP 3577 were employed. The results were compared with human data reported by other investigators as well as with the animal (cat) data obtained earlier in this laboratory. It was found that the conductivity of most of the human tissues tested was significantly higher than that of the animal tissues for the test frequencies. The dielectric constants for human kidney and spleen are higher than the corresponding animal (cat) tissues at frequencies from 10 kHz to approximately 1 MHz and at around 100 MHz. However, the values for liver do not differ significantly between the two species in the same frequency range.

The effect of inhibitors and activators of protein kinase C and phospholipase A, on radiation-induced apoptosis of rat and mouse thymocytes has been studied. It is shown that the apoptosis is prevented by the protein kinase C inhibitor I-(5-isoquinolinylsulfonyl)-2-methylpiperasine dihydrochloride and is potentiated by protein kinase C activator phorbol 12-myristate 13-acetate, calcium ionophore A23187 and concanavalin A. The protein kinase C activators initiate apoptosis in mouse but not in rat thymocytes. The inhibitor of phospholipase Az prevents apoptosis induced by all the factors. The results obtained indicate that both protein kinase C and phospholipase A, are involved in the thymocyte apoptosis.

The effect of the interaction of different types of cells on the interphase death and pycnosis of thymocytes irradiated in vitro was studied. When removed from the thymus suspension of cells with natural killer activity, medullary thymocytes and macrophages did not change the radiation-induced death of cortical thymocytes. On the other hand, postirradiation incubation of cortical thymocytes together with unirradiated thymocytes or with cells of certain other cell lines diminished thymocyte death. Mixing the cell suspensions and changing the incubation medium decreased thymocyte death. All of these results indicate that these cells produce soluble mediators that are toxic to the cells that secrete them. The possible nature of these autotoxic mediators has been studied using inhibitors of arachidonic acid metabolism. Inhibitors of phospholipase A2 or lipoxygenase reduced interphase death markedly, while an inhibitor of cyclooxygenase did not. These data suggest that some lipoxygenase products may serve as autotoxic mediators in the interphase death of thymocytes.

A novel theory is proposed regarding the action of ionizing radiation in the range of very low doses. The basic premise of the theory presented is that the low-dose effect cannot be explained by direct damage to the DNA (as has generally been assumed) and that effects on cellular membranes should be considered instead. Low-dose radiation damaging the plasma membrane decreases the concentration of low-molecular weight compounds (LMWC) inside the cell, which through an unspecific mechanism induces an activation of all enzymes. The mechanism described here has been well substantiated. The changes in the intracellular contents of LMWC and the increase of pH(in) cause chromatin rearrangements, alterations in DNA folding and finally, if the latter are strong enough, expression of various "silent" genes including repair enzyme genes.

The interphase death of irradiated rat thymocytes depends on their concentration during postirradiation incubation. The kinetics of pycnosis and cell death determined with the trypan blue exclusion test in the samples with the highest cell concentration (1-2 x 10(7) cells/ml) is consistent with the data available in the literature, whereas the samples with the lowest concentration (2 x 10(5) cells/ml) undergo almost no pycnosis and death after irradiation with doses up to 50 Gy. On the basis of these results, we suggest a new mechanism of interphase death involving an interaction between irradiated thymocytes and the fraction of thymus cells possessing cytocidal activity. The observed correlation between the cytocidal activity and interphase death of thymocytes from animals of different ages favors our mechanism. It was found that the inhibitors which prevent the conjugation of killer cells and their targets do not influence interphase death, while the substances which block the secretion of cytotoxic factors or their action on the target membrane do protect from interphase death. Thus we suggest that the irradiation activates the killer cells to secrete some cytotoxic factors which induce pycnosis and interphase death of thymocytes.

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