Relationship between the structure of carbonylcyanide phenylhydrazones and inhibition of growth of microorganisms,stimulation of respiration of yeast cells and rat liver mitochondria

The effect of ten derivatives of carbonylcyanide phenylhydrazone on growth of bacteria, yeast and different species of filamentous fungi was investigated. In yeast and mitochondria isolated from rat liver the effect of these derivatives on the respiratory activity was also followed. The relative efficiency of the individual derivatives of earbonylcyanide phenylhydrazone was determined on the basis of the results obtained. It was shown that derivatives, in which the substituent on the benzene ring causes simultaneously an increase of acidity and lipophilicity of the derivative as compared with the non-substituted carbonylcyanide phenylhydrazone (4-trifluoromethoxy-, 3-chloro-, 4-chloro and 3,4-dichloroderivatives) were most effective.     

Cytochemical and biological properties ofMycobacterium bovis BCG

It was the aim of the present communication to find a simple test for a reliable discrimination ofMycobacterium bovis BCG fromMycobacterium tuberculosis. A total of 26 BCG strains, out of them 10 Czechoslovak strains (2 lyophilized cultures of BCG of different batch, 6 strains isolated from abscesses of children after BCG-vaccination and 2 strains from fatal cases after BCG-vaccination) and 16 strains obtained from foreign laboratories, were used. Of the tested characteristics a combination of 3 tests, sensitivity to 1 μg of 2-thiophene carbonylhydrazide (TCH), activity of 3 acylamidases (urease, nicotinamidase and pyrazinamidase) and a quantitative nitrate test, was found to be most advantageous. The Czechoslovak strains ofMycobacterium bovis BCG were fully sensitive to TCH, of the 3 acylamidases mentioned above only urease was positive and nitrate was reduced only little or not at all. On the other hand, strains ofMycobacterium tuberculosis were always resistant to TCH, had positive urease, nicotinamidase and pyrazinamidase and reduced nitrate very intensively.     

Transforming activity of plasmid and chromosomal DNA inEscherichia coli

An auxotrophic strain ofEscherichia coli with therecB recC sbcB genotype was transformed by chromosomal DNA of the prototrophic strain and by plasmid DNA carrying genes for antibiotic resistance (R1drd 19). The donor plasmid DNA obtained by cell lysis in the presence of Triton X-100 and subsequent centrifugation in a caesium chloride-ethidium bromide gradient was shown to have a circulaf molecule and to retain its completeness after penetration into the recipient. Experiments with mixtures or plasmid and chromosomal DNA indicate a competition between these two DNA types during the transformation reaction in the given system.     

Movements and associations of ribosomal subunits in a secretory cell during growth inhibition by starvation

In Chironomus tentans salivary gland cells, the cytoplasm can be dissected into concentric zones situated at increasing distances from the nuclear envelope. After RNA labeling, the newly made ribosomal subunits are found in the cytoplasm mainly in the neighborhood of the nucleus with a gradient of increasing abundance towards the periphery of the cell. The gradient for the small subunit lasts for a few hours and disappears entirely after treatment with puromycin. The large subunit also forms a gradient but one which is only partially abolished by puromycin. The residual gradient which which is resistant to the addition of the drug is probably due to the binding of some large ribosomal units to the membranes of the endoplasmic reticulum (J.-E. Edstrom and u. Lonn. 1976. J. Cell Biol. 70:562-572, and U. Lonn and J.-E. Edstrom. 1976. J. Cell. Biol. 70:573-580). If growth is inhibited by starvation, only the puromycin-sensitive type gradient is observed for the large subunit, suggesting that the attachment of these newly made subunits to the endoplasmic reticulum membranes will not occur. If, on the other hand, the drug-resistant gradient is allowed to form in feeding animals, it is conserved during a subsequent starvation for longer periods than in control feeding animals. This observation provides a further support for an effect of starvation on the normal turnover of the large subunits associated with the endoplasmic reticulum. These results also indicate a considerable structural stability in the cytoplasm of these cells worth little or no gross redistribution of cytoplasmic structures over a period of at least 6 days.     

Genetic definition of a further gene region and identification of at least three different histocompatibility genes in the rat major histocompatibility system

Two new recombinant haplotypes of the rat major histocompatibility system,RT1, have been detected in [LEW.1A (RT1 ^a ) ×LEW.1W (RT1 ^u )] × LEW 1N(RT1 ⁿ ) segregating hybrids. Recombinantr3 carries theRTL1. A region (determining classical transplantation antigens) and theRT1.B region (determining strong mixed lymphocyte reactivity and genetic control of antipolypeptide immune responsiveness) of the RT1^a parent, bur rejects RT1^a skin grafts. Recombinantr4 carries theA andB regions of the RT1^u parent, but rejects RT1^u skin grafts. The two histocompatibility genes detected are allelic to each other. The relevant locus, designated asH-C, maps to theB-region side of theRT1 system and appears to mark a thirdRT1 gene region,RT1.C. Availability of haplotypes r3 andr4 allowed the definition of a histocompatibility locus in theB region,H-B. The products ofH-C, H-B and of the previously describedH-A gene vary in antigenic strength.     

Enzymatic synthesis of β-lactam antibiotics: A thermodynamic background

The equilibrium constants and the respective standard Gibbs energy changes for hydrolysis of some β-lactam antibiotics have been determined. Native and immobilized penicillin amidase (EC 3.5.1.11) from Escherichia coli has been used as a catalyst. The values of standard Gibbs energy changes corresponding to the pH-independent product of equilibrium concentrations (ΔG⁰_c = ? RT ln K_c) have been calculated. The differences in the structure of the antibiotics nucleus hardly ever affect the value of the pH-independent component of the standard Gibbs energy change (ΔG⁰_c) and value of apparent standard Gibbs energy change at a fixed pH (ΔG^0′_c). At the same time, the value of ΔG⁰_c is more sensitive to the structure of the acyl moiety of the antibiotic; when ampicillin is used instead of benzylpenicillin, ΔG⁰_c increases by ～6.3 kJ mol^?1 (1.5 kcal mol^?1). pH-dependences of the apparent standard Gibbs energy changes for hydrolysis of β-lactam antibiotics have been calculated. The pH-dependences of ΔG^0′_c for hydrolysis of all β-lactam antibiotics have a similar pattern. The thermodynamic pH optimum of the synthesis of these compounds is in the acid pH range (pH < 5.0). The breakage of the β-lactam ring leads to a sharp decrease in the ΔG^0′_c value and a change in the pattern of the pH-dependence. For example, at pH 5.0 ΔG^0′_c decreases from 14.4 kJ mol^?1 for benzylpenicillin to ?1.45 kJ mol^?1 for benzylpenicilloic acid. The reason for these changes is mainly a considerable increase in the pK of the amino group of the nucleus of the antibiotic and, as a consequence, a decrease in the component of standard Gibbs energy change, corresponding to the ionization of the system. The thermodynamic potentials of the enzymatic synthesis of semisynthetic penicillins and cephalosporins on the basis of both free acids and their derivatives (N-acylated amino acids, esters) are discussed. It is shown that with esters of the acids, a high yield of the antibiotic can, in principle, be achieved at higher pH values.     

The role of endogenous cytokinins in correlation between cotyledon and its axillary bud and in hypocotyl regeneration of flax

When flax seedlings are decapitated above cotyledons and three days later one of the two cotyledons is removed then the remaining cotyledon stimulates in four to five days growth of its axillary bud. It has been found that content of endogenous cytokinins was higher in the stimulated bud as compared with the other one already 12 h after the cotyledon removal. Flax seedlings decapitated under cotyledons regenerate adventitious buds on thy hypocotyl stump during 5–6 days. The endogenous fytohormonal preparation of this regeneration was investigated in the 20 mm apical part of the hypocotyl stump. Decrease in auxin and increase in gibberellins was already found during the first day after decapitation while the level of cytokinins increased as late as three days after the apex removal.     

Genetic determination of the mitochondrial adenine nucleotide translocation system and ITS role in the eukaryotic cell

On integrating experimental data published previously, the following picture of the mitochondrial adenine nucleotide (AdN) translocation system is being presented: 1. The AdN translocation system serves not only to transport ATP synthesized within mitochondria into the cytosol but also to transport cytosolic ATP into the mitochondria when oxidative phosphorylation is not functioning. 2. The AdN translocator is coded for by nuclear genes and the mitochondrial protein synthesis is not involved in its formation. 3. The AdN translocation system must be preserved and functioning even in cells which could dispense with oxidative phosphorylation. It assures appropriate concentrations of intramitochondrial ATP. 4. The intramitochondrial ATP is required for normal replication of mitochondrial DNA. Tis supports the view that the mitochondrion is a self-replicating semi-autonomous organelle. 5. The appropriate concentration of ATP must be present in mitochondria to make possible cell growth or multiplication. This points to a direct or indirect role of mitochondria in the control of cell proliferation.