The role of oxygen in DNA damage by ionizing particles.

The actual role of oxygen in inactivation mechanism represents still an open problem, especially when Ewing (1998, Am. J. Clin. Oncol.21, 355-361) has shown that oxygen fixation hypothesis cannot be regarded as maintainable more and, on the other side, has argued that the oxygen effect can be hardly a simple consequence of greater reactivity of oxygen radicals. However, the role of oxygen in DSB formation may be more complicated even during the chemical phase of the radiobiological mechanism, as will be shown by computer simulation of the data concerning DSB number dependence on oxygen concentration (for different gas mixtures) and established experimentally by Blok and Loman in 1973 (Radiat. Res.9, 165-245) by irradiating DNA water solutions by gamma radiation.

Mitochondrial generation of oxygen radicals during reoxygenation of ischemic tissues.

Ischemia and reperfusion causes severe mitochondrial damage, including swelling and deposits of hydroxyapatite crystals in the mitochondrial matrix. These crystals are indicative of a massive influx of Ca2+ into the mitochondrial matrix occurring during reoxygenation. We have observed that mitochondria isolated from rat hearts after 90 minutes of anoxia followed by reoxygenation, show a specific inhibition in the electron transport chain between NADH dehydrogenase and ubiquinone in addition to becoming uncoupled (unable to generate ATP). This inhibition is associated with an increased H2O2 formation at the NADH dehydrogenase level in the presence of NADH dependent substrates. Control rat mitochondria exposed for 15 minutes to high Ca2+ (200 nmol/mg protein) also become uncoupled and electron transport inhibited between NADH dehydrogenase and ubiquinone, a lesion similar to that observed in post-ischemic mitochondria. This Ca(2+)-dependent effect is time dependent and may be partially prevented by albumin, suggesting that it may be due to phospholipase A2 activation, releasing fatty acids, leading to both inhibition of electron transport and uncoupling. Addition of arachidonic or linoleic acids to control rat heart mitochondria, inhibits electron transport between Complex I and III. These results are consistent with the following hypothesis: during ischemia, the intracellular energy content drops severely, affecting the cytoplasic concentration of ions such as Na+ and Ca2+. Upon reoxygenation, the mitochondrion is the only organelle capable of eliminating the excess cytoplasmic Ca2+ through an electrogenic process requiring oxygen (the low ATP concentration makes other ATP-dependent Ca2+ transport systems non-operational).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of age, adrenocorticotropin, and dexamethasone on a male-specific cytochrome P450 localized in the inner zone of the guinea pig adrenal.

Guinea pig adrenal microsomes have a capacity for xenobiotic metabolism which is localized to the inner zone, greater in adult males then females, greater in older than in younger males, and suppressed by ACTH. In this paper we show that a cytochrome P450 distinct from P450(21) and P450(17) alpha, which is localized to the inner zone, is male specific, increases with age, and is suppressed by ACTH. This is the first report of a sex-dependent cytochrome P450 in the adrenal. It is immunochemically related to the two members of the P450I family, P450 c and d, neither of which has been reported to be hormonally regulated. The correlation of this cytochrome P450 with the ability of the microsomes to metabolize ethylmorphine suggests that it accounts for this capacity. Whether metabolism of foreign compounds and/or male-specific steroid hydroxylation are its functions in vivo remains to be determined.

A cytochrome P450 immunochemically related to P450c,d (P450I) localized to the smooth microsomes and inner zone of the guinea pig adrenal.

In addition to their capacity for steroid synthesis, guinea pig adrenal microsomes have a well documented ability to metabolize foreign compounds. The capacity for metabolism of foreign compounds is localized to the smooth endoplasmic reticulum-filled cells of the inner zone. However, it has not been clear whether they possess cytochrome P450(s) specific for this function, distinct from the two known steroid hydroxylases, P450(21) and P450(17)alpha. Multiple prominent protein bands in the mol wt range of known cytochrome P450s are seen on sodium dodecyl sulfate gels of guinea pig adrenal microsomes. Most are more intense in smooth microsomes, where the concentration of cytochrome P450 is highest. However, one band (52K) appears unique to the smooth microsomes. This band is also characteristic of microsomes obtained from the inner zone. This protein and two others (54K and 50K) are concentrated in the membrane pellet after carbonate treatment of the microsomes, indicating that they are integral membrane proteins. All three decrease in intensity after treatment of the animals with spironolactone, a compound known to cause depletion of adrenal cytochrome P450s. On Western blots of microsomal proteins the 54K and 50K proteins react with antibodies specific for P450(17) alpha and P450(21), respectively. The 52K protein, characteristic of the smooth microsomes and inner zone, does not react with anti-P450(21) or anti-P450(17) alpha, but does react with polyclonal antibody raised against microsomal cytochrome P450s induced by methylcholanthrene in rat liver (P450c,d). These results suggest that there is at least one additional cytochrome P450 in adrenal microsomes which is immunochemically distinct from P450(21) and P450(17) alpha. Its localization to the smooth microsomes and inner zone microsomes correlates with the high activity for ethylmorphine metabolism detected in these fractions. This suggests that this cytochrome P450, which is immunochemically related to members of the P450I subfamily, may be associated with the ability of guinea pig adrenal microsomes to metabolize foreign compounds.