Modifying role of dietary factors on the mutagenicity of aflatoxin B1: in vitro effect of sulphur-containing amino acids

Sulphur-containing amino acids including some derivatives have been tested for their effectiveness in suppressing the mutagenic activity of aflatoxin B1 in Salmonella typhimurium strains provided with a rat liver activation system. Cysteine and N-acetylcysteine have been found to be most effective in the 2 strains tested (TA100 and TA98). Glutathione (oxidised and reduced forms) has shown partial activity, while cystine and methionine are found to be partially effective only in strain TA100. Inhibition of mutagenicity may be due to interaction of these substances with microsomal enzymes resulting in interference with the formation of ultimate mutagenic species.     

Thromboxane receptor antagonism combined with thromboxane synthase inhibition. 6. 4-substituted 3-pyridinylalkanoic acids.

(3-Pyridinyl)alkanoic acids substituted at the 4-position with an (arylsulfonamido)alkyl group were synthesized and found to behave as platelet thromboxane receptor antagonists (TxRAs) and thromboxane synthase inhibitors (TxSIs). The compounds behaved as agonists at the vascular receptor for thromboxane A₂.     

Mechanism for the opioid-induced twitch potentiations of frog's skeletal muscle

The twitch-potentiating effects of opioids in the frog's skeletal muscle which are naloxone resistant and nonstereospecific were further studied. The rapid kinetics of the onset and of the offset (following washout) of the opioid effect indicates that the site for this action is the surface membrane of the muscle fibre. On the other hand, the lack of any twitch-potentiating effect by naloxone methylbromide, a quaternary derivative of naloxone, suggests that opioids which potentiate the twitch must enter the lipid phase of the membrane to act. Intracellular microelectrode experiments revealed no relation between the opioid effects on membrane electrical events and twitch potentiation. Blocking slow calcium channels with D-600 did not modify the opioid-induced twitch potentiation. The twitch potentiation was antagonized by increasing the extracellular calcium concentration, [Ca2+]o, to 8.64 mM. The effects of closely spaced multiple electrical pulses revealed that the opioids decreased the summated response relative to predrug controls. The results suggest that opioids facilitate the process of excitation-contraction coupling in the frog's skeletal muscle by the release of an additional amount of "trigger calcium" following a single electrical stimulus, thereby generating a potentiated twitch.     

Genotype dependent variation in mycorrhizal colonization and response to inoculation of pearl millet

Summary Genotypes of pearl millet (Pennisetum americanum L. Leeke) were examined for differences in vesicular-arbuscular mycorrhizal (VAM) colonization and response to inoculation. For thirty genotypes tested across three field locations there was a range of mycorrhizal colonization intensity between 25 and 56%. In another experiment with two male-sterile lines, restorer lines and their derived crosses, grown in pots filled with non-sterilized soil there were significant differences between genotypes for colonization by mycorrhiza. This showed hostgenotype dependence for mycorrhizal colonization.Root growth rates, mycorrhizal root length, percentage root colonization and plant growth and P uptake were studied in ten genotypes. A set of 3 genotypes with similar root lengths varied significantly with regard to mycorrhizal root length and the percentage colonization. This supports the suggestion that VAM colonization and spread is dependent on the host genotype. The growth responses differed significantly between the genotypes and they also differed in their responses to P uptake and VAM inoculation. The utility of host-genotype dependent differences in VAM symbiosis in plant breeding is discussed.Journal Article No. 453     

Production of l-Phenylalanine from Starch by Analog-Resistant Mutants of Bacillus polymyxa

p-Fluorophenylalanine-resistant mutants of starch-degrading Bacillus polymyxa ATCC 842, generated by ethyl methanesulfonate mutagenesis followed by incubation with caffeine, overproduced small amounts of l-phenylalanine (l-phe) from starch. A beta-2-thienylalanine-resistant mutant (BT-7) derived from p-fluorophenylalanine mutant (C-4000 FP-4) and resistant to both p-fluorophenylalanine and beta-2-thienylalanine produced 0.5 g of l-phe and 0.15 g of l-tyrosine per liter from 10 g of starch per liter when growing in a minimal medium. trans-Cinnamic acid (CA) was also excreted by both mutants, indicating the possibility of l-phenylalanine ammonia-lyase-induced deamination of l-phe to CA. The amount of l-phe-derived CA detected in BT-7 was less compared with mutant C-4000 FP-4. CA production was induced in the parent only when l-phe was used as a sole nitrogen source. Time of CA production in the two mutants could be delayed by addition of other nitrogen sources, an indication of possible l-phenylalanine ammonia-lyase inhibition or repression. The presence of l-phenylalanine ammonia-lyase in B. polymyxa mutant C-4000 FP-4 was confirmed by assays of cell-free extracts from cells grown in starch minimal medium containing l-phe as the sole nitrogen source. Preliminary studies of the regulation of deoxy-d-arabino-heptulosonate-7-phosphate synthase and prephenate dehydratase in the wild-type strain showed that deoxy-d-arabino-heptulosonate-7-phosphate synthase was subject to feedback inhibition by l-phe, l-tyrosine, and l-tryptophan. Inhibition by each amino acid was to a similar extent singly or in combination at a 0.5 mM level of each amino acid. Prephenate dehydratase was feedback inhibited by l-phe, but not by l-tyrosine or l-tryptophan or both. In the double analog-resistant mutant BT-7, deoxy-d-arabino-heptulosonate-7-phosphate synthase had specific activity similar to that in the wild type, and the enzyme was still subject to feedback inhibition. However, prephenate dehydratase had increased specific activity and it was also insensitive to feedback inhibition by l-phe. The overproduction of aromatic amino acids by BT-7 was thought to be due, at least in part, to deregulation of feedback inhibition of prephenate dehydratase. Chorismate mutase was not subject to feedback inhibition in the wild type and was unaffected in the mutant.     

Purification and general properties of δ-aminolaevulate dehydratase from Nicotiana tabacum L

1. delta-Aminolaevulate dehydratase (EC 4.2.1.24) was purified 80-fold from tobacco leaves and its properties were studied. 2. The enzyme had optimum pH7.4 in potassium phosphate buffer, K(m)6.25x10(-4)m at 37 degrees and pH7.4, optimum temperature 45 degrees and an activation energy of 11100 cal./mole. 3. The enzyme lost activity when prepared in the absence of cysteine, and this activity was only partly restored by the later addition of thiols. Reagents for thiol groups inactivated the enzyme. 4. Mg(2+) was essential for activity, and EDTA and Fe(2+) were inhibitory; Mn(2+) was an activator or an inhibitor depending on the concentration.     

High-performance liquid chromatographic method for the determination of a novel thymidylate synthase inhibitor, AG 331, in human serum

AG 331 is a novel thymidylate synthase inhibitor currently in Phase I clinical trial. To determine the pharmacokinetic parameters of AG 331 in human subjects, a suitable analytical method was developed using high-performance liquid chromatography. Serum and urine samples were prepared using both solid-phase extraction and solvent extraction. Either 4,4′-diaminodiphenyl sulfone or benz[cd]indole-2(1H)-one were used as internal standards for the method. A reversed-phase C₁₈ analytical column completely resolved the drug and internal standard peaks from non-specific substances present in biological matrix. The method was validated for precision, accuracy, and reproducibility in serum and was linear over a concentration range of 50–2000 ng/ml, with a limit of detection of 20.0 ng/ml and a quantifiable limit of 50 ng/ml.