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Biomedical subjects

R H Leach

Publications and source records attributed to R H Leach.

At least 19 recordsLinked to original sources

Relationship between Mycoplasma mycoides subsp. mycoides ('large-colony' strains) and M. mycoides subsp. capri, as indicated by numerical analysis of one-dimensional SDS-PAGE protein patterns.

Twenty-five strains classified as Mycoplasma mycoides subsp. mycoides LC or subsp. capri have been compared by one-dimensional SDS-PAGE of their cellular proteins. A computerized numerical analysis revealed that the protein patterns of all but two aberrant strains formed one large phenon that separated clearly from representatives of the four other members of the 'M. mycoides cluster' at a similarity level (S) of 66% and which remained undivided at up to 78% S. At higher similarity levels, these strains fell heterogeneously into mixed sub-phenons containing strains of both subspecies. Serological comparisons by immunofluorescence largely confirmed the subspecies designations of the test strains, but also showed that some were serologically intermediate between subsp. mycoides and subsp. capri, being cross-reactive with both. These results confirm and enlarge upon those of our earlier studies indicating the protein-pattern inseparability of subsp. capri and subsp. mycoides LC strains and their distinctiveness from the classical M. mycoides subsp. mycoides SC strains and other members of the 'M. mycoides cluster'. As also recognized by other workers, subsp. mycoides LC and subsp. capri strains appear to comprise one large group, wherein those most readily identifiable as either type lie at either end of a serological spectrum that also contains serologically cross-reactive strains. Our observations therefore suggest the lines along which the three groups classified at present within the species M. mycoides (SC and LC strains of subsp. mycoides; subsp. capri) might eventually be reclassified, subject to direct genomic comparisons.

Bacterial Proteins

Numerical analysis of PAGE protein patterns and the taxonomic relationships within the 'Mycoplasma mycoides cluster'.

Twenty-six isolates belonging to the 'Mycoplasma mycoides cluster' have been characterized by one-dimensional SDS-PAGE of their cellular proteins. A numerical classification based on the resulting patterns and using a correlation coefficient revealed four distinct phenons at a similarity (S) level of 70%, comprising: (a) bovine group 7 strains; (b) M. capricolum and F38-like strains; (c) M. mycoides subsp. capri and LC strains ('subsp. mycoides'); (d) M. mycoides subsp. mycoides (SC). At the 75% S level, they could be divided further to give eight phenons. The composition of the clusters at both levels was in good agreement with their previous classification, except for M. mycoides subsp. mycoides LC and M. mycoides subsp. capri, which were clustered in a single phenon at 70% S and could not be clearly separated at 75% S. We conclude that high-resolution SDS-PAGE, combined with computerized analysis of protein patterns, provides an extremely effective approach to the investigation of taxonomic relationships within this group of mycoplasmas.

Bacterial Proteins

Taxonomy of the Mycoplasma mycoides cluster.

Some of the mycoplasmas found in diseases of ruminants, Mycoplasma capricolum, M. mycoides subsp. capri (PG3), bovine group 7 (PG50), strain F38 and strain Y goat show various degrees of relationship to M. mycoides subsp. mycoides (PG1) and to one another, such that they are referred to as the M. mycoides cluster. Studies using serology, DNA homology, isoenzyme analysis and two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) of proteins have been carried out on the type and reference strains of these organisms and some representative strains in an effort to resolve this cluster into well-defined taxa. An ad hoc committee set up by the Subcommittee on the Taxonomy of the Mollicutes examined the known data, but found that it led to conflicting opinions on the classification of the unspeciated strains, partly because of the lack of guidelines on the relative weight to be given to each criterion, and partly because insufficient strains of each member of the cluster had been examined. Although classifications were proposed, total agreement was not achieved on any one proposal, and the committee believes that decisions on the taxonomy would have to await the results of repeat tests, in which several representative strains of each of the six groups would be included with the type strain.

Animals

Construction of the mycoplasma evolutionary tree from 5S rRNA sequence data.

The 5S rRNA sequences of eubacteria and mycoplasmas have been analyzed and a phylogenetic tree constructed. We determined the sequences of 5S rRNA from Clostridium innocuum, Acholeplasma laidlawii, Acholeplasma modicum, Anaeroplasma bactoclasticum, Anaeroplasma abactoclasticum, Ureaplasma urealyticum, Mycoplasma mycoides mycoides, Mycoplasma pneumoniae, and Mycoplasma gallisepticum. Analysis of these and published sequences shows that mycoplasmas form a coherent phylogenetic group that, with C. innocuum, arose as a branch of the low G+C Gram-positive tree, near the lactobacilli and streptococci. The initial event in mycoplasma phylogeny was formation of the Acholeplasma branch; hence, loss of cell wall probably occurred at the time of genome reduction to approximately to 1000 MDa. A subsequent branch produced the Spiroplasma. This branch appears to have been the origin of sterol-requiring mycoplasmas. During development of the Spiroplasma branch there were several independent genome reductions, each to approximately 500 MDa, resulting in Mycoplasma and Ureaplasma species. Mycoplasmas, particularly species with the smallest genomes, have high mutation rates, suggesting that they are in a state of rapid evolution.

Base Sequence

Effect of Mycoplasma pulmonis infection on protein and glycosaminoglycan synthesis of cultured connective tissue cells.

Human synovial cells, fetal skin fibroblasts and rat granulation tissue fibroblasts were experimentally infected with Mycoplasma pulmonis, a species identified as a contamination of cell cultures, and studied for collagen, total protein and glycosaminoglycan synthesis. Hyaluronic acid and sulfated glycosaminoglycan synthesis were stimulated in cultures where the infection reduced cell density, while they were retarded in cultures which had proliferated into higher density than the controls. An extra polypeptide with molecular weight of 20 kD was seen in [35S]methionine-labelled cells. Media of rat granulation tissue cells showed a shift of a 39-42 kD polypeptide to 33-36 kD position in [35S]methionine and [3H]proline labellings. Other minor changes were also noticed. Collagen synthesis or procollagen conversion to collagen were, however, not altered.

Animals

Creutzfeldt-Jakob disease. Failure to detect spiroplasmas by cultivation and serological tests.

Specimens from confirmed cases of Creutzfeldt-Jakob disease (CJD) were examined for evidence of spiroplasma infection. No spiroplasmas or other mycoplasmas were cultivated from brain tissue of 18 cases and no antibodies to several recognised Spiroplasma spp. were detected in sera from 15 patients. These negative cultural and serological results provide no support for published suggestions that CJD brain contains structures morphologically resembling spiroplasmas.

Antibodies

Standard antisera produced in ponies for the identification of bovine mycoplasmas: comparative growth-inhibition results from six laboratories.

Antisera to 10 mycoplasma species of bovine origin were produced in 10 ponies and were distributed for evaluation in growth-inhibition tests at 6 laboratories in Australia, England, Denmark, France, and the United States. Except for a few failures with some antigens produced at the 6 laboratories, the antisera induced large zones of growth inhibition in homologous, but not heterologous, systems. These antisera may be useful as standard reagents for the identification of the bovine mycoplasmas.

Acholeplasma

The plaque-forming factor for mink lung cells present in cytomegalovirus and herpes-zoster virus stocks identified as Mycoplasma hyorhinis.

Previous investigation of the ability of cytomegalovirus and varicella-zoster virus to replicate in a variety of cell lines suggested that both virus types plaqued with high efficiency in mink lung cells. However, many of the virus isolates used appeared to be contaminated with mycoplasma. We now report that the observed cytopathic effect is due to a mycoplasma which grows lytically to high titre in mink lung cells, but is difficult to cultivate in cell-free media. The mycoplasma was plaque-purified and shown to contain DNA with a buoyant density of 1.684 g/ml, with restriction endonuclease patterns identical to the porcine mycoplasma M. hyorhinis. This was confirmed by serological identification.

Animals

An evaluation of a ward pharmacy service.

An evaluation of a ward pharmacy service showed: 1. Its introduction led to a 40-50% reduction in errors and ambiguities of prescribing. 2. It assisted in the safe and more effective use of medicines in hospitals. 3. Medical staff particularly valued the safety checking applied to their prescriptions and the patient's total medication profile, and the opportunity of seeking information about drugs. Nursing staff most appreciated the checking and clarifying elements which assisted during the administration of drugs, the promotion of the most effective use of medicines and the opportunity of discussing problems of drug therapy.

Drug Prescriptions

The distribution of Mycoplasma meleagridis in normal turkey carcases and those affected with air sac disease.

Mycoplasma meleagridis was isolated from both normal and diseased air sacs of turkeys examined and graded for air sacculitis on the evisceration line in a processing plant. It was also cultivated from wing bone cavities of turkeys with severe air sac lesions but not from leg bones or from samples of deep muscle. A further investigation, using material taken randomly from the condemned meat container, confirmed that M meleagridis was recoverable from wing bones.

Air Sacs

Returned-medicines campaign in Birmingham, 1977.

In a returned-medicines campaign lasting 3 weeks 362000 tablets and capsules were returned in 11400 containers from a population of 1.05 million. This yield was considered low. A comparison of hospital admission figures for childhood poisoning before, during, and immediately after the campaign suggests that the publicity and the destruction of unwanted medicines had little preventive value. The quantities of different medicines returned were roughly proportional to the quantities prescribed.

Adolescent

High nonspecific reactivity of normal lymphocytes against mycoplasma-infected target cells in cytotoxicity assays.

Several rat tumor cell cultures were deliberately infected with three species of mycoplasma commonly found as contaminants of cell lines grown in vitro, and the effect of mycoplasma infection on the results of cytotoxicity assays was examined. Lymph node cells and spleen cells from normal animals showed an apparently high spontaneous cytotoxic activity against tumor cells infected with either M. arginini or M. hyorhinis, but the reactivity against cells infected with M. orale was not significantly higher than that against uninfected cells. The high reactivity towards tumor cells infected with M. arginini and M. hyorhinis bore a close resemblence to natural cell-mediated immunity in that spleen cells were much more reactive than lymph node cells, spleen cells from nude mice were as effective as spleen cells from normal mice, and the reaction crossed both strain and species barriers. However, closer examination revealed that the cytotoxic effects were directly caused by depletion of arginine or other essential nutrients from the medium. These findings imply that a cautious approach should be taken when interpreting certain aspects of spontaneous cell-mediated cytotoxicity, and that the greatest care be taken to ensure that the cells used as targets in any cytotoxicity test are mycoplasma-free.

Animals

Use and wastage of prescribed medicines in the home.

A survey of prescribed medicines was carried out in 192 homes, broadly representative of households in England and Wales. The average home had 3.2 containers of such medicines; 56 per cent were in current use, 16 per cent in occasional use, and 28 per cent were never used. One fifth of all oral antibiotics found in the study were wasted. A rough estimate suggests that pound23 million of prescription products (5.6 per cent of the total) are wasted annually in England and Wales.

Drug Prescriptions

Atypical mycoplasmas from sheep in Great Britain and Australia identified as Mycoplasma ovipneumoniae.

Representative strains of "lacy-colony" mycoplasmas isolated from sheep in Great Britain and Victoria (Australia) were classified as M ovipneumoniae following comparison with strain Y98 of this species. The taxonomic description of M ovipneumoniae is extended and Y98 is proposed as the type strain. A brief description is given of the isolation of M ovipneumoniae from sheep in East Anglia.

Animals

Simple transformation method for predicting plasma drug profiles from dissolution rates.

A transformation factor is described which related in vitro drug dissolution from a preparation to the corresponding in vivo plasma drug concentrations. This factor, derived from the dissolution profile and the corresponding in vivo plasma concentration of a single formulation, was used to predict plasma concentration profiles of similar formulations simply from dissolution data.

Methods