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The preparation of iodinated vancomycin and its distribution in bacteria treated with the antibiotic.

ABSTRACT: Vancomycin was radioactively labelled by iodination with (125)I.Iodinated vancomycin was only a little less potent as an antibiotic than vancomycin itself. It was shown, both by chromatography and differential absorption measurements, to combine with acyl-d-alanyl-d-alanine residues. Radioactive vancomycin was used to follow the fate of the antibiotic in bacteria that had been subjected to the least concentration required to inhibit growth. Most of the radioactivity was in the cell walls, although some was found in the membrane fraction. The latter proportion increased during longer incubations with the antibiotic. Pre-formed protoplasts adsorbed very little vancomycin. Mg(2+) removed labelled vancomycin from the mucopeptide of Bacillus licheniformis, but had little effect on the antibiotic adsorbed on Micrococcus lysodeikticus, either in vivo or on previously isolated cell walls. Specific peptide was shown to compete with cell walls for vancomycin and it also extracted from cell-wall samples the labelled compound that had been adsorbed on M. lysodeikticus living cells.

SUBMITTER: Perkins HR

PROVIDER: S-EPMC1185327 | biostudies-other | 1970 Jan

REPOSITORIES: biostudies-other

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Project description:Abstract of associated manuscript: The Bacillus subtilis extracytoplasmic function (ECF) sigma(M) factor is activated by cell envelope stress elicited by antibiotics, and by acid, heat, ethanol and superoxide stresses. Here, we have used several complementary approaches to identify genes controlled by sigma(M). In many cases, expression is only partially dependent on sigma(M) because of both overlapping promoter recognition with other ECF sigma factors and the presence of additional promoter elements. Genes regulated by sigma(M) have a characteristic pattern of induction in response to cell envelope-acting antibiotics as evidenced by hierarchical clustering analysis. sigma(M) also contributes to the expression of the Spx transcription factor and thereby indirectly regulates genes of the Spx regulon. Cell envelope stress responses also include regulons controlled by sigma(W), sigma(B) and several two-component regulatory systems (e.g. LiaRS, YycFG, BceRS). Activation of the sigma(M) regulon increases expression of proteins functioning in transcriptional control, cell wall synthesis and shape determination, cell division, DNA damage monitoring, recombinational repair and detoxification. WT (-van) vs. WT (+van), sigM (-van) vs. sigM (+van), WT (-van) vs. sigM (-van), WT (+van) vs. sigM (+van), WT (-van) vs. spx (-van), WT (+van) vs. spx (+van). Each experiment was conducted at least twice using two independent total RNA preparations. For vancomycin untreated and treated experiments, untreated samples were labeled with Alexa Fluor 555 and treated samples with Alexa Fluor 647. For WT vs. mutant experiments, wild type was labeled with Alexa Fluor 555 and mutants with Alexa Fluor 647. For dye swap experiment, wild-type was labeled with Alexa Fluor 647 and mutant with Alexa Fluor 555. Bacillus subtilis CU1065, WT (-van) vs. WT (+van), sigM (-van) vs. sigM (+van), WT (-van) vs. sigM (-van), WT (+van) vs. sigM (+van), WT (-van) vs. spx (-van), WT (+van) vs. spx (+van)

Project description:Despite their significant impact on public health, antibiotic resistance and size distributions of airborne viable bacteria in indoor environments in neonatal intensive care units (NICU) remain understudied. Therefore, the objective of this study was to assess the antibiotic resistance of airborne viable bacteria for different sizes (0.65-7 µm) in private-style and public-style neonatal intensive care units (NICU). Airborne bacteria concentrations were assessed by a six-stage Andersen impactor, operating at 28.3 L/min. Public-style NICU revealed higher concentrations of airborne viable bacteria (53.00 to 214.37 CFU/m3) than private-style NICU (151.94-466.43), indicating a possible threat to health. In the public-style NICU, Staphylococcus was the highest bacterial genera identified in the present study, were Staphylococcus saprophyticus and Staphylococcus epidermidis predominated, especially in the second bronchi and alveoli size ranges. Alloiococcus otitidis, Bacillus subtiles, Bacillus thuringiensis, Kocuria rosea, and Pseudomonas pseudoalcaligene, were identified in the alveoli size range. In NICU#2, eight species were identified in the alveoli size range: Bacillus cereus, Bacillus subtilis, Bacillus thuringiensis, Eikenella corrodens, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus gordoni. Multi-drug-resistant organisms (MDROs) were found in both of the NICUs. Bacillus cereus strains were resistant to Ampicillin, Cefoxitin, Ceftaroline, and Penicillin G. Staphylococcus cohnii ssp. cohnii was resistant in parallel to ampicillin and G penicillin. Staphylococcus saprophyticus strains were resistant to Ampicillin, Penicillin G, Oxaxilin, and Erythromycin. Results may indicate a potential threat to human health due to the airborne bacteria concentration and their antibiotic resistance ability. The results may provide evidence for the need of interventions to reduce indoor airborne particle concentrations and their transfer to premature infants with underdeveloped immune systems, even though protocols for visitors and cleaning are well-established.

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The preparation of iodinated vancomycin and its distribution in bacteria treated with the antibiotic.

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