Radiochemical assays for creatine kinase and arginine kinase using rapid ion exchange separations

Simple and specific radiochemical assays for the determination of creatine and arginine kinase activities in crude tissue extracts are described. Creatine kinase is assayed by incubation with radioactive creatine and subsequent determination of the radioactivity in creatine phosphate. Creatine and creatine phosphate are separated on DEAE-cellulose ion exchange papers. Arginine kinase is assayed simllarly by using radioactive arginine and separating it from arginine phosphate on short ion exchange columns. An assay for creatine kinase in the direction of creatine formation is also described.     

Augmentation of the antibody response by hapten help: I. Dominant genetic control

The objective of the present work was to characterize those genetic factors that determine susceptibility to “hapten help,” i.e., the augmentation of B-cell responses by hapten-reactive T cells. After mice had been sensitized to the hapten azobenzenearsonate (ABA), one of two approaches was used to assay hapten help. In the first, circumvention of tolerance to low doses of bovine γ-globulin (BGG) was augmented in CBA but not in C57BL/6 mice, as measured by serum anti-BGG antibody after challenge with ABA-BGG. Second, similar strain differences but on a larger scale were demonstrated in the anti-BGG plaque-forming cell (PFC) response of spleen cells from hapten-primed nontolerized mice after challenge with ABA-BGG. Results with the F1-hybrid of a cross between a high responder and a low responder for hapten help demonstrated that high responsiveness is dominant. Experiments with recombinant inbred mice from high- and low-responder progenitor strains suggested that hapten help is associated not with the major histocompatibility complex (H-2) so much as with minor histocompatibility antigens such as H-22 and/or H-24, both of which are on chromosome 7.     

The molecular organization of the beta-globin complex of the deer mouse, Peromyscus maniculatus

Recombinant DNA clones have been isolated that contain 80 kb of the beta-globin complex from the deer mouse, Peromyscus maniculatus. Comparisons of this complex with that from the laboratory mouse, Mus domesticus (with an order 5'-Hbby, Hbb-bhO, Hbb-bhl, Hbb-bh2, Hbb-bh3, Hbb-bl, Hbb-b2 3') highlight organizational trends in the beta-globin complex since the two species diverged. Unlike other mammals studied thus far, the deer mouse possesses three adult genes. Partial sequence analysis indicates that each of the three adult genes is intact and hence may be functional. Hybridization of one of the two Mus pseudogenes, Hbb-bh3, to genomic blots from Peromyscus reveals that it has a homologous counterpart in Peromyscus. Homologous genes to the two gamma-like Mus genes, Hbb-bhO and Hbb-bhl, are also found in Peromyscus. The strong hybridization between the Hbb-bhl genes and significant nucleotide similarity between the Hbb-bhO genes suggest that both pairs are important for the ontogeny of these mice although no known product has been identified for the Hbb-bhO genes. The presence of Hbb-bhO and Hbb-bhl in Peromyscus suggests that the duplication that created this related gene set occurred before the two lineages diverged. A single gene for Hbb-y has been isolated from Peromyscus. The adult region in Peromyscus has undergone significant divergence from the same region in Mus, having three rather than two adult genes, the acquisition of at least 15 kb of extra DNA relative to Mus, and possibly the loss of the Hbb-bh2 pseudogene. The nonadult region of the complex, in contrast, contains the same set of genes apparently distributed over the same amount of DNA as in the Mus beta- globin complex. This observation suggests that the embryonic region of the complex is more evolutionarily stable than the adult region.      

A note on the use of metal species in microbiological tests involving growth media

The feasibility of using traditional growth media for biological testing of metal species, for example as potential microbiocides, was investigated. Significant interactions between both of the representative metal species studied, Cu2+ and FeEDTA, and the test media were found. It is recommended that the use of growth media for tests on metal species should be avoided.     

Two ABA-responsive proteins from pea (Pisum sativum L.) are closely related to intracellular pathogenesis-related proteins

We report here the isolation of cDNAs encoding two abscisic acid-responsive pea (Pisum sativum L.) proteins, ABR17 and ABR18, which are synthesized during late seed development in vivo. Southern blot analyses suggest that ABR17 cDNA corresponds to a single-copy gene, but ABR18 is one member of a family of closely related sequences in the pea genome. The deduced amino acid sequences of ABR17 and ABR18 cDNAs showed similarity to those of the pea disease resistance response proteins, to pathogenesis-related and to stress-induced proteins in other species and to the major birch pollen allergen Betvl.