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Biomedical subjects

W C Noble

Publications and source records attributed to W C Noble.

At least 55 records · Page 3Linked to original sources

Cloning of the gene conferring resistance to mupirocin in Staphylococcus aureus.

A plasmid known to be associated with mupirocin resistance of Staphylococcus aureus has been isolated and a restriction enzyme map constructed. An EcoRI fragment of 4.05 kb from this plasmid has been cloned into an Escherichia coli-Staphylococcus aureus shuttle vector and shown to carry the gene for resistance to mupirocin. The DNA sequence of a small section of the gene has been determined and the derived amino acid sequence compared with a data bank. The amino acid sequence is identical for eight amino acids with the sequence of isoleucyl tRNA synthetase of E. coli. This finding adds to the evidence that mupirocin resistance is the result of a modified isoleucyl tRNA synthetase.

Amino Acid Sequence↗

Tn554 inserts in methicillin-resistant Staphylococcus aureus from Australia and England: comparison with an American methicillin-resistant group.

We have compared methicillin-resistant (Mcr) Staphylococcus aureus isolates from Australia, the UK and the USA with regard to chromosomal inserts of the macrolides-lincosamides-streptogramin B (MLS)-resistance transposon Tn554. The American isolates were known to have a distinctive Tn554 insert, designated insert 6, which was closely associated epidemiologically with the methicillin-resistance phenotype. Southern blots of DNA from Australian and London, UK Mcr isolates were hybridized with a range of probes related to Tn554. The isolates had similar or identical Tn554 inserts, and we consider them to be a single group, designated 'Australondon'. Australondon isolates were compared in detail with a deletion mutant, ANS62, that had lost the methicillin-resistance determinant mec, plus other resistance determinants resident in the mec region of the chromosome, and with an American Mcr isolate containing Tn554 insert 6. The Australondon isolates had three Tn554 inserts. Sequence analysis with the polymerase chain reaction showed that all of these inserts differed from classical Tn554 in that the 3'-terminal residues of the transposons were reverse complements of the usual GATGTA. One of the Australondon inserts, designated 6B, closely resembled Tn554 insert 6 in the sequence of its left flanking chromosomal DNA. This insert was found to abut the deletion from the mec region which results in strain ANS62. We infer that Tn554 insert 6B is part of the mec region of the chromosome in Australondon isolates, supporting the idea that insert 6 of the American isolates is also part of this chromosomal region.

Australia↗

Streptomycin and tetracycline resistance plasmids in Staphylococcus hyicus and other staphylococci.

Plasmids associated with resistance to streptomycin, to streptomycin plus chloramphenicol or to tetracycline in Staphylococcus hyicus isolated from the skin of pigs have been compared, by restriction endonuclease digest patterns, with similar staphylococci from human sources and with published DNA base sequences. Several plasmids from Staph. hyicus have proved to have a very similar structure to those described from Staph. aureus but others appeared very dissimilar. This confirms the opinion that staphylococci from animal skin share a pool of plasmids with those from human skin but may also possess some of quite different structure.

Animals↗

Nasal, axillary, and perineal carriage of Staphylococcus aureus among women: identification of strains producing epidermolytic toxin.

Following two outbreaks of staphylococcal scalded skin syndrome in a maternity unit, 500 pregnant women attending an antenatal clinic were screened for carriage of epidermolytic toxin producing Staphylococcus aureus. Nasal, axillary, and perineal swabs were collected from women whose gestational ages ranged from 12-40 weeks. Isolates of S aureus were purified, phage typed, and tested for methicillin sensitivity and production of epidermolytic toxin. The results showed that 164 (33%) women carried S aureus; of these, 100 (61%) were from the nose and three (2%) from axillae, but 41 (25%) strains were isolated from the perineum alone. Screening for nasal carriage alone will therefore miss 25% of carriers. More than one strain of S aureus was identified in seven of 20 women with multiple site carriage. Three (2%) methicillin resistant strains were isolated during the survey, and five (3%) isolates produced epidermolytic toxin. Phage typing identified 63 (34%) strains as non-typable, but 50% of isolates typed either groups I, II or III, and a further 10% represented varying combinations of these and other phage groups. These results provide baseline information on S aureus in the community, and identification of methicillin resistant and toxin producing strains shows a reservoir of outbreak potential which could become relevant on hospital admission of such a carrier.

Adolescent↗

Antigenic cross-reactivity among isolates of group JK corynebacteria.

Using rocket and rocket-line immunoelectrophoresis and immunoblotting it was demonstrated that a group of antibiotic-resistant bacteria, including several authenticated Corynebacterium jeikeium strains, shared many cross-reacting antigens. Only weak cross-reactivity was demonstrated with representatives of three other skin corynebacteria, C. bovis, C. hofmanii and C. minutissimum or with non-coryneforms. Differences within C. jeikeium are sufficient to permit the use of immunoblotting as an epidemiological tool.

Animals↗

An outbreak of pemphigus neonatorum.

During the summer of 1987, an epidemic of pemphigus neonatorum took place at Guy's Hospital. It involved more than 80 neonates in the maternity unit. Swabs from the umbilical stumps of the babies and from the noses of several attending midwives yielded Staphylococcus aureus of phage-type Group II 3A/3C. Despite an extensive disinfection policy, which included identification and treatment of carriers, the outbreak persisted for 3 months. Final resolution came only after detailed epidemiological research revealed those midwives most likely to be involved. After these had been singled out for further treatment, the outbreak ended. The epidemic strains were later subjected to reverse phage-typing, plasmid profiling and in vivo testing for production of epidermolytic toxin in order to confirm true carriers and cases. Retrospective analysis identified those persons most likely to have been responsible for propagation of the epidemic strain. The exact course of the outbreak was then clarified.

Cross Infection↗

The value of electrophoretic protein patterns for the study of Microsporum canis.

Whole-cell protein analysis of clinical isolates of Microsporum canis by polyacrylamide gel electrophoresis (PAGE) showed small but inconsistent differences in protein patterns. All clinical isolates closely resembled the (-) mating type of Arthroderma otae. A reference strain of A. otae (+) mating type from Japan gave a PAGE pattern which was distinct from those of the clinical isolates. Mating studies showed that the progeny of these reference (+) and (-) mating types had a variety of distinct protein patterns. Protein patterns can therefore distinguish between genetically different strains of M. canis (A. otae) suggesting that clinical isolates have a common clonal origin.

Animals↗

Sterols of fungi responsible for superficial skin and nail infection.

Two groups of fungi isolated from human skin and nail are the dermatophytes Epidermophyton, Microsporum and Trichophyton species and the non-dermatophytes Hendersonula toruloidea and Scytalidium hyalinum. Examination of the sterol composition of these fungi by gas chromatography-mass spectrometry (GC-MS) identified five new sterols from dermatophytes, namely cholesterol, campesterol, episterol, fecosterol and sitosterol. These sterols, with ergosterol and brassicasterol, were also identified from extracts of H. toruloidea and S. hyalinum. Sterol patterns produced by GC-MS with selected ion recording of ten ions were analysed by principal components analysis using the SIMCA statistical package. The dermatophyte genera and species were not differentiated using this method, the similarity in sterol content reflecting the close taxonomic relationship of this group. H. toruloidea and S. hyalinum had similar sterol contents to each other, reinforcing the opinion that these fungi are related, although H. toruloidea form 3 was distinguished from forms 1 and 2.

Arthrodermataceae↗

Diversity of staphylococci exhibiting high-level resistance to mupirocin.

Plasmids mediating high-level resistance to mupirocin (MIC greater than 1000 mg/L) in staphylococci from various sources were studied by restriction endonuclease cleavage. Several patterns were obtained but six plasmids isolated from various Staphylococcus aureus and S. epidermidis strains were indistinguishable. The diversity and spread of these plasmids is illustrated.

Coagulase↗

A comparison of the sterol content of multiple isolates of the Candida albicans Darlington strain with other clinically azole-sensitive and -resistant strains.

The mechanism of azole resistance of Candida albicans NCPF 3310 (the deposited culture of the Darlington strain) has been investigated but never fully explained. Seven isolates of this strain, from various sources, were examined by gas chromatography-mass spectrometry to detect changes in the sterol composition following passage through many laboratories over several years. Five of the seven, including one recently isolated from the patient, were found to be similar to each other in sterol content, containing large amounts of fecosterol. Of the remaining two, one was thought to be a sensitive variant, both produced only small quantities of fecosterol and resembled the normal clinical strains and other azole-resistant strains in sterol content. The sterol composition of the Darlington strain was unique and apparently stable to prolonged in vitro experimentation and passage through the patient.

Azoles↗

Adherence of Staphylococcus aureus to cell monolayers.

Adherence of four strains of Staphylococcus aureus to eukaryotic cell monolayers was assayed with [3H]-thymidine labelled bacterial cells and the results were analysed by non-parametric statistical tests. Adherence to primary (human mesothelial) and semi-continuous (human embryonic lung) cell monolayers was significantly better than to continuous cell lines (HEp2, HeLa and Vero). HEp2 cell monolayers provided the most reliable assay substrate of the continuous cell lines tested. Variation occurred between bacterial culture batches but the assay measured significant differences between adhesion levels of the strains and distinguished between high level (RN92, 8325-4) and low level (Wood46, ISP458) adhering strains. Adherence to different batches of cell monolayers also varied but relative adherence values for strains were similar and the ranking of strains according to adhesion values was unchanged. Potential adhesion mediators have been monitored for their effect on adhesion of a highly adherent strain (RN92) to HEp2 monolayers. Fibronectin, protein A and anti-protein A did not significantly affect adhesion. Lipoteichoic acid caused a significant inhibition of adhesion. With critical statistical analysis to accommodate inherent variations, this assay provides a useful model to study factors involved in adherence of Staph. aureus to eukaryotic cells.

Animals↗

Topical and systemic antibiotics: is there a rationale?

Topical and systemic antibiotic therapy is common in dermatology, yet it is hard to find a rationale for a particular route in some diseases. Any rational choice needs to take efficacy, cost, and the likelihood of antibiotic resistance into account. Aspects of these factors are discussed in relation to the therapy of infectious and noninfectious bacterial skin diseases.

Administration, Topical↗

Use of gas chromatography/mass spectrometry selected-ion recording for sterol content pattern recognition in fungal classification.

Chemotaxonomic studies are frequently limited by the volume of data obtained and the difficulty of reducing this to statistically interpretable results. This paper describes the use of a straightforward procedure for relating the pattern of occurrence of sterols in fungi, analysed by mass spectrometry, to their classification.

Gas Chromatography-Mass Spectrometry↗

Transmissible mupirocin resistance in Staphylococcus aureus.

The spread of two strains of Staphylococcus aureus with high level resistance to mupirocin is described. The resistance proved to be easily transferred to other S. aureus strains by filter mating experiments and on the skin of mice. No plasmid band corresponding to the resistance could be demonstrated by agarose gel electrophoresis or by caesium chloride gradient centrifugation but cleavage of 'chromosomal' DNA from resistant recipients showed bright bands of DNA absent from sensitive controls.

Anti-Bacterial Agents↗

The evaluation of a typing scheme for coagulase-negative staphylococci suitable for epidemiological studies.

A typing scheme was devised for an epidemiological study of infection with coagulase-negative staphylococci in patients undergoing continuous ambulatory peritoneal dialysis (CAPD). The scheme was constructed in four stages suitable for the screening of large numbers of isolates: antibiogram, biotype, phage type, and plasmid profile. The discrimination and reproducibility of the scheme was established by the examination of 50 isolates from 33 consecutive episodes of peritonitis affecting 18 patients. The discrimination of the scheme was 76%, with a reproducibility of 86%. Indistinguishable strains occurred in individual patients only, demonstrating that no cross-infection between patients occurred during the 10-month period of collection of strains, and suggesting that the discriminating power of the scheme was, in fact, much higher. The antibiogram, selected as the first stage of the scheme because it was the simplest and cheapest test, proved to be the most discriminatory stage, providing 60% of the final discriminatory power.

Anti-Bacterial Agents↗