Diagnostic test systems based on the immunoenzyme method in leprosy

Diagnostic test systems for the detection of IgG and IgM to Mycobacterium leprae in the blood sera of leprosy patients and armadillos experimentally infected with M. leprae have been developed on the basis of the indirect immunoperoxidase assay. The possibility has been shown of prognosing the activity of the leprotic process in leprosy patients and the results of the experimental infection of armadillos by the dynamic increase of antibody reactions with the development of the infection.     

Synthesis of new olean-18(19)-ene derivatives from allobetulin

New olean-18(19)-ene triterpenoids were effectively synthesized by the interaction of allobetulin or its acetate with phosphorous oxychloride in refluxing pyridine. The structures of the synthesized 17-chloromethyloleane-18(19)-enes were confirmed by NMR spectroscopy and X-ray analysis.     

Comparison of optimization methods for fed-batch cultures of hybridoma cells

Two methods for the calculation of optimal trajectories for the input variables of a fed-batch culture of hybridoma cells are compared. It pointed out that a gradient method based on Pontryagins' minimum principle based yields a significant better performance with respect to computational effort and the calculated minimum than a dynamic programming approach which has been presented in a previous paper [1] as the most suitable method.     

Ganglioside abnormalities associated with failed neural differentiation in a T-locus mutant mouse embryo

The T-locus on mouse chromosome 17 contains a number of mutations that disrupt cellular differentiation and embryonic development. Because of their purported role in neuronal differentiation and brain development, gangliosides were studied in mouse embryos homozygous for two T-locus mutations: T and twl. Mice homozygous for the dominant T mutation die from failed mesodermal differentiation in the notochord, whereas mice homozygous for the recessive twl mutation die from failed neural differentiation in the ventral portion of the neural tube. No major ganglioside abnormalities were found in T/T mutant embryos at Embryonic Day 10 (E-10). In contrast, E-11 twl/twl mutants expressed a marked deficiency of the tetrasialoganglioside GQ1. Since this ganglioside migrates with GQ1b in three different thin-layer solvent systems, it may have the same structure as GQ1b. To gain insight into regional distribution, gangliosides were examined in head regions and body regions of normal (+/+) E-11 embryos. The ganglioside composition of these regions was the same as that of the whole embryo, with GM3 and GD3 comprising about 75% of the total ganglioside distribution. Moreover, N-acetylneuraminic acid was the only sialic acid species detectable in the E-10 and the E-11 embryos. These findings indicate that N-acetylneuraminic acid-containing gangliosides are synthesized actively in E-10 and E-11 mouse embryos and also suggest that the GQ1 deficiency in the twl/twl mutants is closely associated with failed neural differentiation.     

In vitro modulation using a synthetic antioxidant of the stimulus-induced formation of cyclic AMP

The in vitro treatment of membranes isolated from different rat organs with a water-soluble synthetic antioxidant has resulted in the change of basal and stimulus-induced adenylate cyclase activity. It is believed that the antioxidant effect is realized rather at the level of signal transfer from activated receptor to adenylate cyclase than at the level of agonist-receptor interaction.     

Production of tetanolysin preparations and characteristics of their properties

As the result of the study of tetanolysin-producing Clostridium tetani strains, their populations have been found to be markedly heterogeneous with respect to the hemolytic activity of clone cultures. On the basis of normal and dialyzed cultures of selected variants with maximum activity the preparations of tetanolysin have been obtained, and their hemolytic activity and antigenic properties have been studied. Antihemolytic rabbit sera have also been obtained and characterized. Partially purified preparations of tetanolysin with high hemolytic activity have been obtained by the fractionation of C. tetani dialyzed cultures with ammonium sulfate.     

Cloning and expression of the Erwinia chrysanthemi asparaginase gene in Escherichia coli and Erwinia carotovora

A genomic library of Erwinia chrysanthemi DNA was constructed in bacteriophage lambda 1059 and recombinants expressing Er. chrysanthemi asparaginase detected using purified anti-asparaginase IgG. The gene was subcloned on a 4.7 kb EcoRI DNA restriction fragment into pUC9 to generate the recombinant plasmid pASN30. The position and orientation of the asparaginase structural gene was determined by subcloning. The enzyme was produced at high levels in Escherichia coli (5% of soluble protein) and was shown to be exported to the periplasmic space. Purified asparaginase from E. coli cells carrying pASN30 was indistinguishable from the Erwinia enzyme on the basis of specific activity [660-700 units (mg protein)-1], pI value (8.5), and subunit molecular weight (32 X 10(3]. Expression of the cloned gene was subject to glucose repression in E. coli but was not significantly repressed by glycerol. Recombinant plasmids, containing the asparaginase gene, when introduced into Erwinia carotovora, caused increased synthesis of the enzyme (2-4 fold higher than the current production strain).