Biological method of identifying antibiotic substances at an early screening stage

A biological method was used in addition to the chemical methods of identification in the screening programme of new antibiotics. The method consists of evaluation of the effect of the crude antibiotic preparations on microbial forms resistant to various antibiotics. The efficiency of the biological method is shown. It provides more complete and rapid characterization of the properties of the new antibiotics and their rough identification at early screening stages.     

Testing the sterility of injection preparations of penicillin series antibiotics]

The efficiency of 3 variants of the method for determination of microbial flora was compared on the injection preparation of potassium benzylpenicillin artificially infected with Staph. aureus 209P and the spores of Bac. subtilis ATCC 6633 in different doses and with different amounts of the preparation in the vials. The procedure of the preparation dissolution in the vial with the thioglycol medium containing penicillinase proved to be most effective. The microbe detection amounted to 100 per cent. The procedure was less labour- and time-consuming since addition of penicillinase to each vial with the thioglycol medium was excluded. The risk of the medium occasional infection with microbial flora during the assay was decreased.     

Some effects of chromium toxicity on bush bean plants grown in soil

Summary Chromium applied to a noncalcareous soil at 50 ppm did not decrease yields of bush beans (Phaseolus vulgaris L. var Improved Tendergreen), but when EDTA (ethylenediamine tetraacetic acid) was added with it, it did. Very little Cr was present in leaves. In solution culture 10^-5 M Cr and higher were toxic. With solution culture the highest level of Cr in leaves was about 30 ppm and in general there was a decreasing gradient in Cr from roots to stems to leaves. EDTA had less effect in solution cultures on Cr toxicity because the Cr was already in solution. Chromium toxicity decreased cation levels in plants.     

Actinomyces streptomycini mutant blocked in streptidine biosynthesis]

Mutant 170 not capable of forming streptidine and streptomycin was obtained using chemical mutagenes. This mutant can produce streptomycin only with suplementation of exogenous streptidine. Experiment with labeled C14-streptidine showed its specific incorporation in streptidine moiety of streptomycin molecule.     

Electron microscopic study of the localization of penicillin acylase in E. coli cells]

The paper describes the studies on definition of penicillinacylase localizations in the cells of E. coli with the help of ferritin labeled immune sera and electron microscopy. Both the intact cells and the cells treated with the substances affecting the cell wall intactness were used. The study showed relation between penicillinacylase and the surface structures of the cell, i.e. the cell wall and the periplasmic areas. It was found that penicillinacylase got into the environmental medium with the splitted cell fragments which corresponded to the general mechanism of excretion of large high molecular compounds by gramnegative organisms.     

Determination of the penicillin acylase activity of E. coli cultures using a method of gas-liquid chromatography]

Penicillinacylase activity was determined in E. coli by the product of benzylpenicillin destruction, i.e. phenylacetic acid formed under the effect of the enzyme. The determination was performed on a chromatograph. The immobile phase consisted of 10 per cent of ethylenglycol edipate on chromosorb A, modified with 2 per cent H3PO4. Nitrogen was used as the gaseous carrier. The method is rapid and handy for mass testing of the cultures with a purpose of detecting penicillinacylase-producing strains. It provided reliable determination of penicillinacylase in the cultures producing simultaneously beta-lactamase, another penicillin-destroying enzyme.     

Polynucleotide sequence relationships among Ent plasmids and the relationship between Ent and other plasmids.

Deoxyribonucleic acid-deoxyribonucleic acid hybridization studies reveal that the plasmids coding for the production of heat stable and heat labile enteroxtoxins of Escherichia coli, regardless of their origin, have a majority of their polynucleotide sequences in common, but are not related in any significant way to those plasmids coding for the synthesis of only ST toxin. The heat stable and heat labile plasmids also share a significant degree of their polynucleotide sequences with plasmids of the FI and FII incompatibility groups, but not with R factors belonging to the I, N, W, P, or X incompatibility groups.     

Molecular cloning of an Escherichia coli plasmid determinant than encodes for the production of heat-stable enterotoxin.

A conjugative plasmid, ESF0041 was isolated from an enterotoxigenic strain of Escherichia coli from calves. ESF0041 was found to be 65 x 10(6) daltons in mass of a member of the F incompatibility complex. Acquisition of ESF0041 by E. coli K-12 was invariably associated with the capacity to produce heat-stable (ST) enterotoxin. ESF0041 and pSC101 deoxyribonucleic acids were cleaved with EcoRI, and the fragments were ligated with polynucleotide ligase. Transformation of E. coli K-12 with the ligation mixture led to the isolation of an ST+ clone. Further analysis of the plasmid deoxyribonucleic acid from this clone showed that a structural gene(s) associated with ST biosynthesis had been isolated as a 5.7 x 10(6)-dalton ESF0041 fragment in pSC101. In turn, 5.7 x 10(6)-dalton fragment was ligated to a multicopy COLE1 derivative, RSF2124, so that toxin synthesis was amplified about threefold.