Radioimmune, radiobinding and HPLC analysis of 2-5A and related oligonucleotides from intact cells

The enzyme (2-5A synthetase) which synthesizes ppp(A2'p)nA where n=2 to 4 (collectively referred to as 2-5A) is widely distributed in a variety of cells and tissues in amounts which increase response to interferon and vary with growth and hormone status. 2-5A activates a nuclease which inhibits protein synthesis. The non-phosphorylated 'core' of 2-5A ((A2'p)nA, n=2 to 4) can inhibit DNA synthesis and cell growth. Here we describe convenient and sensitive radioimmune (RI) and radiobinding (RB) assays for core and 2-5A. In combination with more satisfactory high performance liquid chromatography (HPCL) methods using reverse-phase C18 columns, these assays have been used to detect core and 2-5A in crude extracts from interferon-treated cells. The novel 2-5A synthetase products NAD2'p5' A2'p5'A and A5'p45'A2'p5'A2'p5'A (ref. 13), which can also be detected using the RB assay, were not found in significant amounts. The natural occurrence of core has not been described previously.     

Nicotinamide administration alters the activities of hepatic microsomal mixed function oxidases

Summary Systemic action of nicotinamide significantly alters the activities of hepatic drug metabolizing enzymes. Male rats injected with nicotinamide have reduced levels of cytochrome P450, demethylases and aniline hydroxylase. The changes appear to be sex-dependent since in the case of female rats activities of p-nitroanisole-o-demethylase and aniline hydroxylase are enhanced whereas cytochrome P450 content remains unaltered.Authors gratefully acknowledge the encouragement given by Professor A.S. Paintal during the course of this investigation. One of us (J.K.B.) is grateful to the Council of Scientific and Industrial Research, New Delhi, for the award of a research fellowship.