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V Braun

Publications and source records attributed to V Braun.

At least 253 records · Page 14Linked to original sources
PubMed1981

[Permeability barrier of bacterial cell envelopes as cause of resistance to antibiotics (author's transl)].

The uptake of antibiotics into bacterial cells depends on the structures and mechanisms involved in general transport of substrates. Because of their different molecular architecture, the distinct layers of the cell envelope differ in permeability. The cell-wall of gram-positive bacteria, which is simply structured, is therefore more permeable to a series of antibiotics than is the gram-negative cell-wall which is built like other biological membranes. The energy dependent uptake of many antimetabolic antibiotics in managed by the selective transport systems of the cytoplasmic membrane. In the case of antibiotics, which are not analogous in structure with metabolites, the dependence of the drug on active transport is less obvious. The uptake of antibiotics may even be influenced by the target-site of the drug in the cytoplasm. An example is the accumulation of aminoglycoside antibiotics which is initiated by binding of the drugs to the ribosome. The autoinduction of aminoglycoside uptake is associated with an increased transport of polyamines. The broad variety of modes of resistance, based on permeability, results from the large number of components involved in the uptake of antibiotics. Resistance of bacteria can depend on alterations of every layer of the cell envelope. The existence of a capsule can cause resistance, as well as the specific loss of protein components of the outer membrane, which are involved in the uptake of special substrates. The exclusion of antimetabolites by the loss of the cognate transport system yields well defined resistance. By contrast, resistance towards a wide variety of unrelated antibiotics results from the uncoupling of energization and transport phenomena. Resistance based on reduced uptake becomes even more complex when the modification of the target site alters the accumulation of antibiotics. Far beyond common mechanisms is the ability of cells to actively excrete antibiotics. Far beyond common mechanisms is the ability of cells to actively excrete antibiotics that have already penetrated. The induction of selective export systems for tetracyclines is not dependent on activities of cell-borne transport systems but is a consequence of the acquisition of resistance plasmids.

Anti-Bacterial Agents↗
PubMed1980

Excretion of a protease by Serratia marcescens.

Excretion of an extracellular protease of Serratia marcescens ATCC 25419 occurred during logarithmic growth and was highest (per cell) when cultures reached the stationary growth phase. Production of the extracellular protease was induced by leucine or casein in minimal medium or by growth in tryptone-yeast medium. In the late stationary phase an intracellular protease activity accumulated which was also observed in mutants with very low extracellular protease activity. The excreted protease was the dominant protein in the growth medium. The protease was purified to homogeneity by column chromatography on Bio-Gel P-100 and on DEAE-cellulose. Quantitative amino acid analysis revealed the absence of sulfur-containing amino acids. The enzyme consists of one polypeptide chain. A molecular weiht of 51,000 and 55,000 was estimated using polyacrylamide gel electrophoresis and chromatography on Bio-Gel P-100 respectively. The enzyme cleaved only N-alpha-benzoyl-DL-lysine-and-arginine-nitroanilides but not the corresponding leucine or tyrosine derivatives nor a set of di- and tripeptides.

Amino Acids↗
PubMed1980

Penetration of colicin M into cells of Escherichia coli.

A new class of colicin M-tolerant mutants of Escherichia coli K-12 was isolated. The mutants exhibited an unusually high tolerance in that they were unaffected by colicin titers of 10(6). The tolerance was confined to colicin M. It was mapped at a locus called tolM, which is close to rpsL. The following gene order was determined: aroE, tolM, rpsL, cysG. The tolerance could be caused by a defect in the uptake of colicin M or by a mutation at the site of action. Insensitive tonA and tonB mutants became sensitive to colicin M upon treatment by osmotic shock, whereas the tolM mutants remained insensitive. Trypsin rescue experiments showed that the tonB-dependent uptake of colicin M required energy like the other tonB-related transport processes. When bound to energy-depleted cells, colicin M prevented adsorption of phage T5. The receptor became accessible to the phage when the cells were energized, except in tonB mutants. These data suggest that the function controlled by the tonB gene is required for the translocation of colicin M from its initial binding site at the tonA-coded receptor protein to the target.

Biological Transport, Active↗
PubMed1980

Iron transport in Escherichia coli: uptake and modification of ferrichrome.

During the transport of iron as ferrichrome complex into cells of Escherichia coli K-12, the ligand was modified and excreted into the medium. The rate of the formation of the modified product corresponded with the rate of iron transport. The modified product showed a decreased affinity for ferric iron and did not serve as an effective iron ionophore. After all of the ferrichrome had been converted, the modified product was taken up into the cell in an iron-free form. The uptake of ferrichrome and of the modified product depended on the transport system specified by the tonA and tonB genes. The modified product could be converted back into ferrichrome by mild acid or alkaline hydrolysis. One mole of acetate was released per mole of ferrichrome. It is proposed that one N-hydroxyl group of ferrichrome is acetylated to explain the low affinity for iron as the N-hydroxyl groups form the ligands for iron (III). A weak ester linkage by which the acetyl group is covalently bonded would account for the easy hydrolysis. The iron-free form of ferrichrome, deferri-ferrichrome, was also rapidly converted when incubated with cells with a functional transport system. It is therefore likely that iron is released from ferrichrome by reduction before modification takes place. The conversion of the ligand could be a mechanism by which cells rid themselves of a potentially deleterious ligand for iron in the cytoplasm. A possible role in ferrichrome transport is discussed.

Acetates↗
PubMed1980

Genetic control of hydroxamate-mediated iron uptake in Escherichia coli.

Cells of Escherichia coli can derive iron from a variety of chelators (siderophores) in addition to enterochelin, the catechol derivative excreted by many enteric bacteria. The genetic control of hydroxamate siderophore utilization was investigated in mutants of E. coli K-12 selected for resistance to lethal agents which adsorb to the tonA protein of the outer membrane (albomycin, colicin M, and phages T5 and phi80). Many of the mutants were unable to utilize hydroxamate siderophores as an iron source. This phenotype was termed Fhu, for ferric hydroxamate uptake. Mutants carrying lesions in the tonA region of the chromosome were studied and fell into several types. Members of one class had lost some or all of the tonA receptor protein's functions in that they were resistant to the lethal agents and unable to utilize ferrichrome and its analogs, although able to respond to the hydroxamate rhodotorulic acid (FhuA phenotype). Other mutants were unable to utilize any of the hydroxamate siderophores tested and were resistant to albomycin, althogh many were sensitive to the other lethal agents (FhuB phenotype). Members of these classes lacked ferrichrome-mediated iron uptake. Strains carrying transposon Tn10 insertions in fhuA (previously termed tonA) lacked the 78,000-molecular-weight outer membrane protein previously described; insertions in fhuB retained this protein and had no detectable change in outer membrane composition. Three-point transduction crosses revealed the gene order to be pan-fhuA-fhuB-metD in the min 3.5 region of the chromosome map. Complementation analyses with F' merodiploid strains showed that fhuA and fhuB comprise separate transcription units which are both required for utilization of ferrichrome. Response of diploid strains to rhodotorulic acid suggested the existence of a third gene, fhuC, required for utilization of this siderophore, but not ferrichrome. It is suggested that fhuB encodes a transport component in the cytoplasmic membrane that is necessary for the uptake of all hydroxamate siderophores following their receptor-mediated passage across the outer membrane.

Bacterial Proteins↗
PubMed1980

[The value of conventional chest radiography in the interpretation of pulmonary hemodynamics in mitral valve disease (author's transl)].

Standard biplane chest roentgenograms of 107 patients suffering from acquired mitral valve disease were independently interpreted by two radiologists. Fluoroscopy was performed for detection of mitral valve calcification. The effective pressures in the pulmonary vascular bed were not known at the time of the film reading. Using the most valuable roentgen signs (enlargement of the left atrium, valvular calcification, configuration of the right ventricular outflow tract, size of the central pulmonary arteries and degree of pulmonary interstitial edema), each case was graded from zero to nine points. The two readers widely agree in the classification of the roentgen findings in most cases. There is a strong positive correlation between the radiographic changes and the pulmonary artery mean pressure.

Evaluation Studies as Topic↗
PubMed1979

Iron supply of Escherichia coli with polymer-bound ferricrocin.

Uptake of ferric iron from ferricrocin was studied in Escherichia coli using a polymer-coupled ferricrocin that was unable to penetrate into the cell. Ferricrocinyl polyethylene glycol succinate (Mr 7000 -- 8500) promoted growth of E. coli K-12 AB2847 aroB under iron-limiting conditions. In iron-starved cells, uptake of 55Fe could be demonstrated; the amount of iron accumulated amounted to 10% of that observed with free ferricrocin. The iron supply by ferricrocin bound to polyethylene glycol was strictly dependent upon the functions expressed by the tonA and the tonB genes, as was the iron uptake promoted by free ferricrocin. Polymer-bound ferricrocin protected cells against colicin M and phage T5 by competition for the common tonA-coded outer membrane receptor protein. In addition, the rate of iron transport via the negatively charged ferricrocinyl succinate was as fast as via the neutral ferricrocin molecule. No ligand was found associated with the cells. Penetration of chelator beyond receptor is not necessary for siderophore-mediated iron uptake. It is concluded that sufficient amounts of iron can be released from the polymer complex to satisfy growth requirements.

Biological Transport↗
PubMed1979

Relationship between steps in 8-anilino-1-naphthalene sulfonate (ANS) fluorescence and changes in the energized membrane state and in intracellular and extracellular adenosine 5'-triphosphate (ATP) levels following bacteriophage T5 infection of Escherichia coli.

The addition of bacteriophage T5 to anaerobic, fermenting cells of Escherichia coli B or K-12 in the presence of 8-anilino-1-naphthalene sulfonate (ANS), N-phenylnaphthyl-1-amine (NPN), or dansyl ethylamine causes the fluorescence of these probes to rise in two steps, the first occurring immediately upon addition, the second delayed by 6 min. The conditions necessary for observing this phenomenon are defined (cell density, probe concentration, substrate, absence of an electron acceptor, multiplicity of infection, growth, and harvesting conditions). The magnitudes of the first and second steps in fluorescence are dependent upon the multiplicity of infection; the timing of the steps is not. The first step correlates with a breakdown in the potassium or rubidium permeability barrier of the cells, and it occurs either aerobically or anaerobically, with fermentable or nonfermentable substrates. The second step occurs only with cells that are without an available electron acceptor, are fermenting, and which have a functional membrane-bound, Ca2+-dependent adenosine triphosphatase (ATPase). The results are consistent with disturbance of energization of the cell membrane by the membrane-bound ATPase at the time of the second step in fluorescence. No changes in the intracellular level of adenosine 5'-triphosphate (ATP) was seen, whereas the extracellular level increased sharply, starting 3--6 min after phage addition. The quantity of ATP found in the medium by 30 min after infection amounted to about four times the amount present inside the cells at the time of infection. The quantity and rate of efflux of ATP was similar under aerobic and anaerobic conditions.

Adenosine Triphosphate↗
PubMed1979

Protein II influences ferrichrome-iron transport in Escherichia coli K12.

Ferrichrome-promoted iron uptake in Escherichia coli K12 is strictly dependent upon the tonA gene product, a 'minor' outer membrane protein. By selection for mutants of E. coli resistant to phages which require 'major' outer membrane proteins as receptors, strains with pronounced protein deficiencies were constructed. Such strains were tested for anomalous behaviour of ferrichrome transport. No significant differences in iron uptake were detected in E. coli K12 strains with markedly reduced amounts of protein I. However, a reduction in the initial velocity (up to 40%) was observed in E. coli deficient in outer membrane protein II. This difference was only evident when cells were grown under iron-starvation conditions; it was abolished when cells were grown in rich medium. Kinetic parameters for ferrichrome transport were determined for maximum velocity but for Km; double reciprocal plots showed a biphasic nature, probably attributable to a limited number of outer membrane binding sites and to the multi-component nature of the ferrichrome-iron transport system.

Bacterial Proteins↗
PubMed1979

Uptake and conversion of the antibiotic albomycin by Escherichia coli K-12.

The antibiotic albomycin is transported into cells of Escherichia coli K-12 by the same uptake system as the iron-supplying ferrichrome complex. The iron-complexing hydroxamate moieties of albomycin and ferrichrome are structurally similar. During the phase of rapid iron uptake the chelators were not found in the cells. In order to understand the antibiotic activity of albomycin, it was labeled in the hydroxamate with tritium and in the presumed antibiotically active area with radioactive sulfur. While the tritium label was not retained by the cells, part of the sulfur label was taken up and concentrated 500-fold within the cell. The sulfur was not incorporated into proteins or nucleic acids since it was recovered as a low molecular weight component. Gel filtration on Bio-Gel P-2 revealed one tritium-labeled and two sulfur-labeled cleavage products in the incubation medium. We conclude that albomycin is actively transported via its ferrichrome-like portion into the cells and that the growth-inhibitory moiety is released by hydrolysis intracellularly and remains there.

Anti-Bacterial Agents↗
PubMed1979

Accelerated adsorption of bacteriophage T5 to Escherichia coli F, resulting from reversible tail fiber-lipopolysaccharide binding.

A dual specificity for phage T5 adsorption to Escherichia coli cells is shown. The tail fiber-containing phages T5(+) and mutant hd-3 adsorbed rapidly to E. coli F (1.2 x 10(-9) ml min(-1)), whereas the adsorption rate of the tail fiber-less mutants hd-1, hd-2, and hd-4 was low (7 x 10(-11) ml min(-1)). The differences in adsorption rates were due to the particular lipopolysaccharide structure of E. coli F. Phage T4-resistant mutants of E. coli F with an altered lipopolysaccharide structure exhibited similar low adsorption for all phage strains with and without tail fibers. The same held true for E. coli K-12 and B which also differ from E. coli F in their lipopolysaccharide structures. Only the tail fiber-containing phages reversibly bound to isolated lipopolysaccharides of E. coli F. Infection by all phage strains strictly depended on the tonA-coded protein in the outer membrane of E. coli. We assume that the reversible preadsorption by the tail fibers to lipopolysaccharide accelerates infection which occurs via the highly specific irreversible binding of the phage tail to the tonA-coded protein receptor. The difference between rapid and slow adsorption was also revealed by the competition between ferrichrome and T5 for binding to their common tonA-coded receptor in tonB strains of E. coli. Whereas binding of T5(+) to E. coli K-12 and of the tail-fiber-less mutant hd-2 to E. coli F and K-12 was inhibited 50% by about 0.01 muM ferrichrome, adsorption of T5 to E. coli F was inhibited only 40% by even 1,000-fold higher ferrichrome concentrations.

Adsorption↗
PubMed1979

[The direct detection of sinus nodal potentials in man by means of the signal-averaging technique (author's transl)].

In 13 patients in the age range of 34 to 70 years (mean: 53 +/- 10 years), during diagnostic His-bundle investigations, registrations from the sinus nodal area were analyzed with a signal-averaging computer. In 12 patients we were able to get reproducible per-P-potentials with a sino-atrial conduction interval between 56 and 227 ms. The normal sequence of the excitation process from the atrium to the AV node together with the bundle of His and to the ventricles in all patients, and the reproducibility of the signal-averaged pre-P-potentials, which were well outlined from the atrial potential with a reasonable conduction interval, suggest that these signal-averaged pre-P-potentials reflect the sinus nodal activity rather than an ectopic focus. In 5 of 11 comparable cases there was a good correlation between the signal-averaged sino-atrial conduction intervals and those calculated with the extrastimulus technique. In 2 patients this correlation was moderate. The signal-averaging process may be impaired by coarse random noise, frequent atrial or ventricular extrasystoles, and by fluctuations of the baseline of the sinus nodal lead. The fact that the registrations are performed during normal undisturbed sinus rhythm offers a considerable advantage over the extrastimulus technique.

Action Potentials↗
PubMed1978

Changes in fluorescence of 8-anilino-1-naphthalene sulfonate after bacteriophage T5 infection of Escherichia coli. Initial fluorescence rise coincides with onset of rubidium efflux.

Escherichia coli cells pre-loaded with 86Rb+ begin to lose 86Rb+ immediately after phage T5 addition. The loss proceeds with negative-exponential (first-order) kinetics for up to approximately 15 min after phage addition. The constant which characterizes the rate of loss increases with increasing numbers of infecting phage per cell. It is known that anaerobic, fermenting cells of E. coli show a two-step increase in 8-anilino-1-naphthalene sulfonate (ANS) fluorescence upon infection with bacteriophage T5; the first rise begins immediately upon phage addition, the second 6 min later. The onset of 86Rb+ release, therefore, is correlated with the first fluorescence rise with respect to timing and response to the multiplicity of infection.

Aerobiosis↗
PubMed1978

Functional interaction of the tonA/tonB receptor system in Escherichia coli.

Host range mutants of phage T1 (T1h), which productively infected tonB mutants of Escherichia coli, were isolated. The phage mutants were inactivated by isolated outer membranes of E. coli in contrast to the wild-type phage, which only adsorbed reversibly. For the infection process, the tonB function is apparently only required for the irreversible adsorption of the phage T1, but not for the transfer of the phage DNA through the outer membrane and the cytoplasmic membrane of the cell. Mutants of the tonA gene expressing normal amounts of outer membrane receptor proteins were isolated and found to be partially sensitive to phage T5 and resistant to the phages T1 and T1h, colicin M, and albomycin and unable to take up iron as a ferrichrome complex. One tonA mutant remained partially sensitive to T5, colicin M, and albomycin and supported growth of T1h (not of T1) with the same plating efficiency as the parent strain. Only a small region of the tonA receptor protein seems to function for all the very different substrates. A newly isolated host range mutant of T5 (T5h) adsorbed faster to tonA(+) cells than did wild-type T5 and infected tonA missense mutants resistant to wild-type T5. The interplay of the tonA with the tonB function was observed with phage T5 infection, although T5 required only the tonA receptor. Ferrichrome inhibited plaque formation of T5 only when plated on tonB mutants. Adsorption of T5 to cells in liquid medium was influenced by ferrichrome as follows: complete inhibition by 0.1 muM ferrichrome with tonB mutants, not more than 35% inhibition by 1 to 100 muM ferrichrome with the tonB(+) parent strain in the presence of glucose as energy source, and 90% inhibition by 1 muM ferrichrome with partially starved parent cells. We conclude that there exist different functional states of the receptor protein that depend on the energy state of the cell and the tonB function. The latter seems to be required only for translocation processes with outer membrane proteins involved.

Adsorption↗