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Biomedical subjects

F E Young

Publications and source records attributed to F E Young.

At least 109 records · Page 6Linked to original sources

Soluble macromolecular complexes involving bacterial teichoic acids.

Cell wall and membrane teichoic acids from several bacteria formed soluble complexes with polysaccharides and bovine plasma in alkyl alcohol solutions. Polysaccharides which contain different monomeric units and anomeric configurations complexed with the teichoic acids, suggesting that the interaction is relatively nonspecific. Teichoic acids complexed glycogen or bovine plasma albumin in 50 to 97% ethanol solutions. The macromolecular association between teichoic acids and polysaccharides or proteins was independent of teichoic acid size over a threefold molecular weight range. Glycerol phosphates or an acid hydrolysate of teichoic acid would not complex to either glycogen or bovine plasma albumin in ethanol. The optimal interaction between glycogen and the Bacillus subtilis lipoteichoic acid occurred between pH 4.5 and 8.2. The ability of teichoic acids to bind polysaccharides and proteins in moderate dielectric constant solvents suggests that these polymers may serve as complexing agents for hydrophilic molecules found in membranes.

Bacillus subtilis↗

Stability of pathogenic colony types of Neisseria gonorrhoeae in liquid culture by using the parameters of colonial morphology and deoxyribonucleic acid transformation.

This investigation describes the surveillance of the colonial stability of the pathogenic type 1 from the gonococcal strain F62 to the nonvirulent types 3 and 4 in different liquid media. The maintenance of the colony types was monitored by the parameters of colonial morphology and deoxyribonucleic acid-mediated transformation. During growth in a complex medium, Mueller-Hinton broth, only 46.7% of the gonococcal population remained as type 1 after 12 h. The greatest change in the type 1 colony-forming units correlated with the decline in viable count. The conversion process could not be prevented by the continual maintenance of the gonococcus in logarithmic growth. The frequency of transformation from PRO(minus) (proline) to PRO(plus) was proportional to this decrease in type 1 colony-forming units. In contrast to Mueller-Hinton medium, the chemically defined minimal medium Gonococcal Genetic Medium (GGM) was capable of maintaining approximately 90% of the gonococcal population in the type 1 colonial form after 16 h of growth, despite a decrease in the viable count. Although the percentage of type 1 appeared to remain constant in GGM, the apparent transformation frequency increased approximately 24-fold from 0 to 12 h of growth. GGM appears to stimulate or maintain competence, as evidenced by an eightfold increase in transformation when cells are exposed to deoxyribonucleic acid in GGM as compared to Mueller-Hinton.

Culture Media↗

Characterization of a virulent bacteriophage for Bacillus subtilis (var. amyloliquefaciens).

A phage, designated PBA12, has been isolated from the soil and found to be virulent on Bacillus subtilis (var. amyloliquefaciens). PBA12 has a large cylindrical head that is 100 nm long and 35 nm in diameter and a tail that is 200 nm in length. The phage contains double-stranded DNA and demonstrates chloroform sensitivity. The processes of both adsorption and replication appear to be slow and inefficient.

Bacillus subtilis↗

Development of a defined minimal medium for the growth of Neisseria gonorrhoeae.

This investigation describes the development of a solid and a liquid medium (Gonococcal Genetic Medium; GGM) which support the rapid growth of 41 gonococcal clinical isolates and laboratory strains with a minimum number of nutritional components. The complete medium contains minimal salts, eight amino acids, two nitrogen bases, vitamins, coenzymes, key metabolic intermediates, and some miscellaneous components. Results indicate that GGM can be modified and simplified even further than we described. In liquid GGM, several gonococcal strains grew logarithmically after a 2- to 3-h lag period with generation times ranging from 72 to 115 min, reaching optical densities of 175 to 320 Klett units in the presence of seven amino acids and in the absence of a CO(2) atmosphere. The development of a solid and a liquid defined minimal medium such as GGM should greatly broaden the avenues of experimentation for biochemical genetic studies with N. gonorrhoeae, especially gonococcal genetic transformation. N. gonorrhoeae can be classified into eight major and minor phenotypic groups, depending on its growth responses on GGM to just five amino acids: cysteine and cystine, arginine, proline, isoleucine, and serine. Such results demonstrate the feasibility of using GGM as a simple, sensitive, rapid probe for investigating the epidemiological patterns of gonorrhea.

Amino Acids↗

Polyelectrolyte nature of bacterial teichoic acids.

Several physicochemical properties of the teichoic acid of Bacillus subtilis 168 have been determined. The teichoic acid partial specific volume was found to be 0.57 ml/g. The apparent weight-average molecular weight of the polymer was 24,800. Sedimentation was strongly dependent on solvent. The sedimentation coefficient of the teichoic acid was found to have a value of s(20.w) (0) = 1.90S. In dilute buffers and distilled water, the teichoic acid possessed a rigid rod or extended conformation. Salts induced a loss of secondary structure in the polymer, resulting in a random coil configuration. Salt-induced structural changes in the teichoic acid were determined by viscosities, ultraviolet difference spectra, and inhibition of precipitation with concanavalin A. Divalent cations such as Mg(2+) had little effect on the teichoic acid structure. The salt-induced structural changes were reversible, as evidenced by return of the original properties upon dialysis of the teichoic acid against water. Sodium chloride inhibited the adsorption of bacteriophage ø25 to B. subtilis cell walls. Teichoic acid conformation may have a significant influence on the physiology of bacteria.

Adsorption↗

Autolysis in Staphylococcus aureus: preferential release of old cell walls.

The kinetics of release of old versus new cell wall in two strains of Staphylococcus aureus were studied during autolysis. In both strains the autolytic enzyme is an amidase. Cells were double labeled with (3)H and (14)C, and the distribution of radioactivity in the cell walls was monitored during autolysis. In all cases the rate of release of steady-state lable from peptidoglycan was significantly higher than that of pulse label. Identical results were obtained with whole cells or isolated cell walls. The results suggest that in S. aureus the old cell wall is preferentially released during autolysis.

Amidohydrolases↗

Inhibition of transformation of Bacillus subtilis by heavy metals.

Mercuric ions, as well as organomercuric ions and cadmium ions, can inhibit deoxyribonucleic acid-mediated transformation in Bacillus subtilis 168 without decreasing the viability of the total population. Differences in the inhibition of transformation by mercuric ions are identifiable on a temporal and concentration dependence basis. Sensitivity to low concentrations (9.2 x 10(-8) M) appears early in the uptake of deoxyribonucleic acid before the transformed markers have become insensitive to deoxyribonuclease. Resistance to "low concentrations" of Hg(2+) is kinetically indistinguishable from the requirement for magnesium in the transformation process. This inactivation is not reversed by the mercury-binding compound glutathione. Sensitivity to mercuric ions at a higher concentration (5.52 x 10(-7) M) occurs after the donor deoxyribonucleic acid has become insensitive to deoxyribonuclease. These complex interactions between mercuric ions and the process of transformation are discussed.

Bacillus subtilis↗

Regulation of bacterial cell walls: turnover of cell wall in Staphylococcus aureus.

The cell wall of Staphylococcus aureus was shown to undergo turnover during exponential growth. The rate of turnover, about 15% per generation, was identical for both cell wall polymers, peptidoglycan and teichoic acid. Both the old and newly synthesized wall material appeared to undergo turnover at similar rates. The rate of turnover followed first-order kinetics until more than 90% of the original wall was lost. Cell wall turnover was completely blocked under conditions of unbalanced synthesis known to inhibit cellular autolysis, e.g., addition of chloramphenicol. Cell wall turnover was shown to occur in a number of different strains of S. aureus and appears to be widely distributed in this species.

Bacterial Proteins↗

Active transport of D-alanine and related amino acids by whole cells of Bacillus subtilis.

Whole cells of Bacillus subtilis transported d-alanine and l-alanine by two different systems. The high-affinity system (K(m) of 1 muM and V(max) of 0.6 to 0.8 nmol/min per mg of protein) was specific for the two stereoisomers of alanine. The low-affinity system (K(m) of 10 muM for l-alanine and 20 muM for d-alanine and glycine) had a V(max) of 5 to 12 nmol/min per mg of protein. This system transported glycine, d-cycloserine, and d-serine, in addition to d- and l-alanine. Azide inhibited the uptake of these amino acids and caused the efflux of d-alanine from preloaded cells. These data suggest that transport of these amino acids is energized by the electron transport chain.

Alanine↗

Bacteriophage interference in Bacillus subtilis 168.

Strains of Bacillus subtilis lysogenic for temperate bacteriophage SPO2 inhibit the development of bacteriophage phi1. After infection by bacteriophage phi1, DNA and RNA synthesis in the lysogenic host terminates, culminating in cell death. Bacteriophage SPO2 also prevents the production of bacteriophage phi105. Mechanisms for these two types of bacteriophage interference are discussed.

Bacillus subtilis↗

Characterization of temperate bacteriophages of Bacillus subtilis by the restriction endonuclease EcoRI: evidence for three different temperate bacteriophages.

Temperate bacteriophages of Bacillus subtilis were characterized according to host range and digestion of the bacteriophage genome by endonuclease EcoRI. The three bacteriophages, phi3T, SPO2, and phi105, were all heteroimmune, and the DNA digests showed dissimilar patterns by agarose-ethidium bromide gel electrophoresis.

Bacillus subtilis↗

Transduction in Bacillus subtilis by bacteriophage SPP1.

Lysates of the virulent bacteriophage SPP1 were shown to be capable of mediating generalized transduction. Suppressible mutants of this bacteriophage (sus) were capable of transduction at a lower multiplicity of infection than virulent SPP1. Linkage analysis demonstrated that bacteriophage SPP1 transduced segments of the genome equal in size to that transferred by SP10. This bacteriophage should be useful in analyzing the regions of the genome where PBS1 appears to give anomalous results.

Bacillus subtilis↗