Interaction of polypeptide neurotoxins with a receptor site associated with voltage-sensitive sodium channels

Anthopleurin A, a polypeptide toxin from the Pacific sea anemone Anthopleura xanthogrammica, enhances persistent activation of voltage-sensitive sodium channels by the alkaloid toxins veratridine and batrachotoxin with K0.5 = 20 nM. This effect is inhibited by depolarization. There is a close correlation between enhancement of sodium channel activation and block of [125I]scorpion toxin binding by unlabeled scorpion toxin, sea anemone toxin II from Anemonia sulcata, and Anthopleurin A, indicating that these three polypeptide toxins interact with a common receptor site in modifying sodium channel function. Photo-activable derivatives of scorpion toxin label a single Mr approximately 250,000 polypeptide chain at the polypeptide toxin receptor site. Labeling is blocked by unlabeled scorpion toxin or depolarization and is not observed in variant neuroblastoma clones, which lack sodium channels. These results identify a protein component of the polypeptide toxin receptor site of voltage-sensitive sodium channels.     

Purification of a hybrid plasminogen activator protein

Recombinant DNA technology has been employed to produce a hybrid gene in which the kringle and serine protease domains of tissue plasminogen activator are linked to the heavy-chain Fd region of a fibrin-specific antibody. The hybrid gene is co-expressed with antibody light chains. This communication describes a purification procedure for the hybrid protein, involving affinity and ion-exchange chromatography. The purified hybrid protein has been used in vivo and in vitro clot lysis experiments and has been shown to be effective at clot dissolution.     

A sensitive, rapid ferricyanide-mediated toxicity bioassay developed using Escherichia coli

A need for rapid toxicity techniques has seen recent research into developing new microbiological assays and characterising their toxicity responses using a range of substances. A microbiological bioassay that determines changes in ferricyanide-mediated respiration for toxicity measurement (FM-TOX) shows particular promise. The development and optimisation of an improved FM-TOX method, incorporating novel features, is described using Escherichia coli as the biocatalyst. Omission of an exogenous carbon source, used in previously described FM-TOX assays, substantially improves the assay sensitivity. In addition, the development of a two-step procedure (toxicant exposure followed by determination of microbial respiratory activity) was found to enhance the inhibition of E. coli by 3,5-dichlorophenol and four other toxicants, compared to single-step procedures. Other assay parameters, such as the ferricyanide concentration, exposure times and optical density of the biocatalyst were also optimised, sometimes based on practical aspects. Toxicity tests were carried out using the adopted technique on both organic and inorganic toxicants, with classic sigmoid-shaped dose-response curves observed, as well as some non-standard responses. IC₅₀ data is presented for a number of common toxicants. The optimised assay provides a good foundation for further toxicity testing using E. coli, as well as the potential for expanding the technique to utilise other bacteria with complementary toxicity responses, thereby allowing use of the assay in a range of applications.     

The organo-metal initiated polymerization of vinyl ketones—II : The triethylaluminium initiated polymerization of methyl isopropenyl ketone

A study has been made of the kinetics and mechanism of the triethylaluminium initiated polymerization at 0° of methyl isopropenyl ketone (3-methyl-but-3-ene-2-one) in toluene and tetrahydrofuran. The reaction between triethylaluminium and methyl isopropenyl ketone gives rise to a yellow “ate”-complex which rearranges to the initiating species. A rapid production of linear trimer is followed by a slow polymerization giving rise to a bimodal molecular weight distribution. A coordinate mechanism for the polymerization is proposed.     

Continuous Bundles of C*-Algebras with Discontinuous Tensor Products

For each non-exact C*-algebra A and infinite compact Hausdorffspace X there exists a continuous bundle B of C*-algebras onX such that the minimal tensor product bundle AB is discontinuous.The bundle B can be chosen to be unital with constant simplefibre. When X is metrizable, B can also be chosen to be separable.As a corollary, a C*-algebra A is exact if and only if A Bis continuous for all unital continuous C*-bundles B on a giveninfinite compact Hausdorff base space. The key to proving theseresults is showing that for a non-exact C*-algebra A there existsa separable unital continuous C*-bundle B on [0,1] such thatA B is continuous on [0,1] and discontinuous at 1, a counter-intuitiveresult. For a non-exact C*-algebra A and separable C*-bundleB on [0,1], the set of points of discontinuity of A B in [0,1]can be of positive Lebesgue measure, and even of measure 1.2000 Mathematics Subject Classification 46L06 (primary), 46L35(secondary).     

The use of microorganisms with broad range substrate utilisation for the ferricyanide-mediated rapid determination of biochemical oxygen demand

The feasibility of replacing oxygen with a synthetic electron acceptor in microbial catabolism was investigated as a rapid method for the determination of biochemical oxygen demand (BOD). Microorganisms known for their broad range organic substrate utilisation were investigated. It was shown that Trichosporon cutaneum, Pseudomonas putida and Bacillus licheniformis could utilize the ferricyanide ion as an alternative electron acceptor, in place of oxygen, for the catabolic oxidation of a range of simple organic compounds. The biochemical reactions were monitored by measuring the amount of microbially produced ferrocyanide using amperometry at a Pt disk microelectrode. Catabolic degradation efficiencies approaching those of the conventional 5-day assay were achieved in 1 h. BOD₅ equivalent values for a range of simple organic solutions were determined for each of the microorganisms. The effect of increased incubation time and the choice of appropriate calibration standards for rapid BOD assays were also considered.