Genetically engineered V79 Chinese hamster cell expression of purified cytochrome P-450IIB1 monooxygenase activity

Chinese hamster V79 fibroblasts, frequently used as target cells in short-term tests for mutagenicity, do not possess measurable monooxygenase activity; in particular, enzymatic oxidation of testosterone (T) cannot be demonstrated. If these V79 cells, however, had been transfected with the cDNA-encoding rat liver cytochrome P-450IIB1 under control of the SV40 early promoter, they stably expressed monooxygenase activity. These so-called SD1 cells then oxidatively metabolized T at a rate of 27 pmol/mg protein/min, converting it to 16 alpha- and 16 beta-hydroxy-T as well as 4-androsten-3,17-dione as sole metabolites in a ratio of 1.1:1.0:1.6. The regio- and stereoselective conversion of T by SD1 cells, as well as the quantitative distribution of the metabolites, corresponds well with the results reported for pure cytochrome P-450IIB1 in a reconstituted system.     

Distribution and fate of aliphatic and aromatic hydrocarbons in Antarctic fauna and environment

With the depletion of oil resources in more accessible areas, those of remote regions are being considered or indeed are now being exploited. In many of these regions, especially the polar ones, little is known of the effects such exploitation will have on the environment. But it is known that the ecosystems are often subject to great stress by natural climatic conditions and additional burdens imposed by man may have catastrophic environmental effects. South Georgia, a sub-Antarctic island, has a history of industrial activity mainly concerned with whaling operations that peaked around 1925–1935 but has since declined to virtually nothing. Studies of the ecology of the area provided a unique opportunity to assess the long-term effects that such activities had on the ecosystem. Off the whaling stations a considerable amount of waste material, including fuel oil, was released into the bays and inevitably some of this material was deposited in the sediments. Chemical evidence in the form of both paraffinic and aromatic hydrocarbons still persists in the sediments. The implications of this persistence in relation to the possible influence of the low temperature conditions are discussed. The superficial sediments, marine biota and terrestrial plants, which since 1965 have returned virtually to a pristine state, contain hydrocarbons essentially similar to unpolluted areas around the coast of Britain. Relatively high levels of carcinogenic/mutagenic polynuclear aromatic hydrocarbons in surface sediments suggests a world-wide background of abiogenic hydrocarbons probably disseminated by airborne transport. This appears to indicate that contamination reaches even remote parts of the world in relatively undiminished quantities.     

Reduced calcium and inhibition of protein kinase C mimic the enhancement of ornithine decarboxylase activity of prolactin in Ambystoma tigrinum tissues

We previously reported that prolactin (PRL) could increase the activity of ornithine decarboxylase (ODC) in liver slices taken from larval tiger salamanders (Ambystoma tigrinum). This action of the hormone was inhibited by oxytocin (OT), the calcium ionophore A23187, and diacyglycerol (DG) and was duplicated by 10 microM verapamil (VML), a calcium channel blocker. Here, we expand these results to show that 1) a higher dose of VML (50 microM) produces an additive effect with PRL; 2) addition of small amounts of calcium (0.1 mM) to the liver culture medium blocks PRL action; 3) neither nifedipine (NIF), a different type of calcium channel blocker, nor EDTA alter PRL action; and 4) gossypol, a reported inhibitor of protein kinase C, mimics PRL action. Additionally, we show that PRL increases ODC activity in tiger salamander tail skin in vitro, a tissue previously demonstrated to be a PRL target tissue in this species. The same set of treatments which we have shown to modify PRL effects on ODC in liver slices affects PRL action in the tail skin in a parallel manner. Thus, the mechanism whereby PRL enhances ODC activity appears to be the same in both these tissues. These results are discussed in conjunction with the findings from similar studies using mammalian tissues in an attempt to assess the current picture of the mechanism of PRL action and the possible role of inositol phospholipid turnover, calcium, and protein kinase C in the action of this hormone.