Isolation of Tyr- mutants of facultative methylotrophic Pseudomonas sp. M

A method for isolation of Tyr- mutants of facultative methylotrophic bacteria Pseudomonas sp. M which possess two tyrosine synthesis pathways is presented. The method is based on the two-step blocking of the tyrosine synthesis: the first step of the supplementary pathway of synthesis from phenylalanine, the second being the main pathway from 4-hydroxyphenylpyruvate.     

Assay of DNA-RNA hybrids by S1 nuclease digestion and adsorption to DEAE-cellulose filters.

A fast and accurate assay procedure for DNA-RNA hybrids is described in which exhaustive digestion of unhybridized DNA with S1 nuclease is followed by binding of hybrids to filter discs of DEAE-cellulose. The digested DNA can be efficiently washed from the filters so that background levels of 0.1-0.2% of input tracer DNA can be achieved, in contrast to the much higher (approximately 1-5%) backgrounds obtained using TCA precipitation procedures. Short duplexes, as small as 36 nucleotides in length, which are inefficiently bound to hydroxyapatite, are quantitatively bound to the DEAE-cellulose filters.     

Immunological diagnosis of Mycoplasma pneumonias]

A complex of immunological cell tests with M. pneumoniae antigen (the lymphocyte blast-cell transformation test, the allergic neutrophil alteration test) was carried out in order to establish the correlation between the results of positive seroconversion and the sepcific immunological reactivity of lymphoid cells in pneumonia patients. Mycoplasmic cutireactive allergen, when used for the accelerated diagnosis of mycoplasmic pneumonia in humans, was shown to be specific and safe. Cuti-allergic tests with mycoplasmic allergen allowed to diagnose mycoplasmic pneumonia at early stages (beginning from days 5--7), which ensures the possibility of indicating etiotropic treatment to patients in due time.     

Ultrastructural demonstration of NAD-pyrophosphorylase activity in mouse liver nuclei.

Ultrastructural demonstration of NAD-pyrophosphorylase activity (E.C.2.7.7.1) in isolated mouse liver nuclei was investigated with the use of an electronhistochemical procedure based on the precipitation of pyrophosphate ions with lead ions under conditions permitting simultaneous ATPase inhibition by formaldehyde/ethanol prefixation. In isolated mouse liver nuclei activity of NAD-pyrophosphorylase was found in nucleoli, in interchromatin granules, coiled bodies and strand-like structures in nucleoplasm.     

Genotypical characteristics of the immunological Vi-antibody maturation depending on the effect of Vi-antigen deposit in the body

The authors carried out a comparative analysis of the changes in the antibody synthesis and their immunological maturation in the immunization with various doses of the Vi-antigen of linear rats differing by the capacity to deposit this antigen. August rats with a high capacity to deposit the Vi-antigen (in comparison with Wistar rats in which this capacity was less) were characterized by an increased level of highly avid antibodies. The interlinear differences were found in the character of the changes in the antibody synthesis depending on the immunizing dose of the antigen. Genotypical peculiarities of the immunological maturation of the antibodies were demonstrated. The authors suppose that the described phenomenon was genetically determined and referred to the factors determining the individual immunological reactivity. In the choice of the optimal immunizing doses of the antigens the authors recommend complex determination of the antibody level and the degree of their avidity.     

Distribution of ACTH-like immunoreactivity in rat brain and gastrointestinal tract.

Immunocytochemistry reveals ACTH-like immunoreactivity to reside not only in the pituitary but also in central nerves and in central nerves and in antral gastrin cells. In all probability, the central nerves store a peptide identical with or closely resembling true ACTH. Their pattern of distribution is, in some regions, similar to that of enkephalin-immunoreactive nerves. The antiserum demonstrating ACTH-like immunoreactivity in central nerves and in antral gastrin cells is directed towards the COOH-terminal part of the hormone. A peptide corresponding to this part, the corticotrophin-like intermediate peptide (CLIP) is manufactured by the pars intermedia of the pituitary. CLIP is devoid of adrenocortical activity, but has recently been shown to possess insulin-releasing activity. The occurrence of CLIP-like peptides in antral gastrin cells may indicate a role for such peptides in the gastrointestinal regulation of insulin release. The simultaneous occurrence of enkephalin-like and ACTH-like immunoreactivity in the antral gastrin cells is of particular interest since a large precursor molecule, containing both the enkephalin and the ACTH sequence has recently been identified.