Resistance of Pseudomonas to Quaternary Ammonium Compounds: II. Cross-Resistance Characteristics of a Mutant of Pseudomonas aeruginosa

Tube dilution experiments showed that benzalkonium chloride (BC)-resistant mutants of Pseudomonas aeruginosa grown in the presence of 1,000 mug of BC per ml were at least 20 times more sensitive to polymyxin B and colistin sulfate than the BC-sensitive (BCS) parent strain. BCS cells selected for resistance to 500 mug of polymyxin B per ml remained sensitive to BC. There was little difference in the amount of carbenicillin, gentamicin sulfate, or rifampin needed to prevent growth of either the BCS or BC-resistant (BCR) strains. Growth of BCR cells was inhibited by ethylenediaminetetraacetate at a concentration of 400 mug/ml or less, whereas the BCS strain grew at ethylenediaminetetraacetate levels of 10,000 mug/ml. Phenylmercuric acetate and thimerosal inhibited growth of BCR and BCS cells at concentrations of 10 mug/ml or less. BCR cells were cross-resistant to >1,000 mug/ml concentrations of five other quaternary ammonium compounds, including three with C(16) alkyls and two with alkyl groups of shorter length. The BCS strain was also resistant to >1,000 mug/ml concentrations of the three quaternary ammonium compounds with C(16) alkyl groups but, in addition to BC, was inhibited by 200 mug/ml levels or less of the two quaternary ammonium compounds containing alkyl groups of less than 16 carbon atoms.     

Glucosylation of Teichoic Acid: Solubilization and Partial Characterization of the Uridine Diphosphoglucose:Polyglycerolteichoic Acid Glucosyl Transferase from Membranes of Bacillus subtilis

Polyglycerolteichoic acid:glucosyl transferase (TAG transferase), one of the three enzymes involved in the pathway leading to the glucosylation of teichoic acid in Bacillus subtilis 168, was investigated. During the early stages of the growth of B. subtilis, TAG transferase is predominantly a soluble enzyme found in the cytoplasm. As growth proceeds, the amount of soluble enzyme decreases and the proportion of insoluble, membrane-bound TAG transferase increases, reaching a maximal value at the close of the logarithmic phase. Data are presented which suggest that these are two forms of the same enzyme, or have some common component. The effects of chaotropic agents, such as sodium trichloroacetate and sodium perchlorate, on the cytoplasmic membrane were also studied. These data show that such compounds can effectively remove the TAG transferase from the membrane in a water-soluble form. A study of some of the physical properties of this solubilized enzyme suggests that there is little difference between the two forms of the enzyme. Experiments are described which indicate that the glucosyl transfer by both the membrane-bound and soluble enzymes is not mediated by lipids.     

EVIDENCE FOR THE OCCURRENCE AND DISTRIBUTION OF INORGANIC POLYPHOSPHATES IN VERTEBRATE TISSUES

Evidence for the occurrence of polyphosphates having apparent chain-lengths ranging from less than 10 to over 5000 orthophosphate units has been found in adult brain as well as in a number of other mammalian tissues which have been examined. There appears to be three times as much polyphosphate in rat brain as there is in rat liver. Adult rat brain appears to contain at least 15 μg of phosphorus as polyphosphate per g of fresh tissue. In addition, neural polyphosphate is extremely labile to catabolic degradation after death whereas hepatic polyphosphate is relatively more stable. The nature of this inorganic polymer was elucidated through use of the technique of ³¹P nuclear magnetic resonance spectroscopy. Further structural evidence was obtained by application of the isotopic dilution technique to ³²P-labelled neural polyphosphate mixed with an abiotically prepared inorganic polyphosphate. The conditions and rates of hydrolysis of the biological polyphosphate and a non-biological polyphosphate were comparable.     

Salt injury to plants with special reference to cations versus anions and ion activities

Summary In contrast with the toxicities of sulfate and chloride salts added to substrates, the anions SO₄ and C1 were not injurious when accumulated without leaf burning by cotton and tomato plants from atmospheres enriched with SO₂ or HC1 gases. The foregoing results are discussed in terms of cationenzyme interactions which appear to represent at least a major cause of salt toxicity. Although anions are largely unreactive with enzymes it has long been observed that chloride salts in soil solutions are far more toxic than sulfate salts. Five of seven species have shown nearly equal growth repressions on substrates with 100 me/1 of C1 salts versus 200 me of SO₄ salts, each added as 50 per cent Na. The ion activities of the two solutions were equal and the sum of cations in the plant saps were similar. The osmotic differentials (average about 10 atm) between the expressed tissue fluids and these substrate solutions were remarkably uniform within species. It is projected that the downward transport of salts via the phloem provides for root concentrations which supply ions to the xylem and thereby control the uptake of substrate salts.     

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.     

Mutation in Saccharomyces cerevisiae Conferring Streptomycin and Cold Sensitivity by Affecting Ribosome Formation and Function

A cold-sensitive, streptomycin-sensitive mutant of Saccharomyces cerevisiae accumulates a 28S ribonucleoprotein particle when grown at low temperature. This particle contains 17S ribosomal ribonculeic acid which is degraded when exposed to ribonuclease. The particle does not serve as a precursor to 60 and 40S ribosomal subunits nor is it turned over when growth is allowed to resume at the permissive temperature; rather it is only diluted by growth. That streptomycin sensitivity (allelic with cold sensitivity) is ribosomal is evidenced by the inhibition of protein synthesis in vitro by streptomycin and the binding of labeled streptomycin to the mutant but not the parental 40S ribosomal subunit.