PubMed Health⌕ Search

Biomedical subjects

I R Poxton

Publications and source records attributed to I R Poxton.

At least 19 recordsLinked to original sources
PubMed1992

Nosocomial diarrhoea due to a single strain of Clostridium difficile: a prolonged outbreak in elderly patients.

An outbreak of diarrhoea occurred in an acute geriatric ward of a hospital (A). It affected six patients initially and was found to be due to a single strain of Clostridium difficile. There was little evidence for asymptomatic carriage of this strain or others in the hospital patients. The following three months saw an increase in the number of symptomatic cases of C. difficile disease in two peripheral hospitals, B and C. Patients had been moved from the outbreak ward to these hospitals. Of 18 cases in hospital B all 15 isolates saved for typing were of the 'outbreak' strain. Of three cases occurring in hospital C, only one was the 'outbreak' strain (a relapsed patient who was part of the original episode). There were seven further cases in geriatric and medical wards of hospital A. All six typed isolates were also the outbreak strain. By chance, four isolates from hospital A and four from hospital B pre-dating the outbreak were also available for typing and seven of these were found to be identical to the outbreak strain. This suggests that one strain of C. difficile was endemic in geriatric and medical facilities on two sites and was responsible for nosocomial diarrhoea over at least one year. The problems of optimal management and infection control in this situation are discussed.

Aged↗
PubMed1992

Frequencies of lipopolysaccharide core types among clinical isolates of Escherichia coli defined with monoclonal antibodies.

Mouse monoclonal antibodies (MAbs) specific for the lipopolysaccharide (LPS) core types R1, R2, and R3 of Escherichia coli and a cross-reactive MAb that binds to the LPS core of almost all E. coli were used in ELISA to determine the frequency of cores resembling R1, R2, and R3 in strains of E. coli isolated from clinical samples (blood and urine specimens) and from the feces of asymptomatic individuals. Of the 180 wild-type isolates, 123 were assigned to R1 core type, 14 to R2, and 18 to R3. Twenty-five wild-type E. coli isolates could not be assigned to a particular core type and may have either an R4 or K12 core or a previously unrecognized core type. R1 core type was associated with O types 1, 4, 6, 8, and 18 and with K1 or K5 capsules. R3 was associated with O15. O75 isolates could be of either R1 or R2 core type.

Antibodies, Monoclonal↗
PubMed1992

Production and characterisation of mouse monoclonal antibodies reactive with the lipopolysaccharide core of Pseudomonas aeruginosa.

Monoclonal antibodies (MAbs) to the core antigen region of lipopolysaccharide (LPS) of Pseudomonas aeruginosa were produced from mice immunised with whole cells of heat-killed rough mutants of Pseudomonas aeruginosa expressing partial or complete core LPS. MAbs were screened in an enzyme-linked immunosorbent assay (ELISA) against three different antigen cocktails: S-form LPS from three P. aeruginosa strains, R-form LPS from six P. aeruginosa strains and, as a negative control, R-form LPS from Salmonella typhimurium and Escherichia coli. Selected MAbs were subsequently screened against a range of extracted LPS and whole cells from both reference strains and clinical isolates of P. aeruginosa. The antibodies were also screened in ELISA against whole-cell antigens from other Pseudomonas spp. as well as strains of Haemophilus influenzae, Neisseria subflava and Staphylococcus aureus. Five MAbs reacting with the core component of P. aeruginosa LPS were finally selected. Two of these, MAbs 360.7 and 304.1.4, were particularly reactive in immunoblots against unsubstituted core LPS, including that from O-antigenic serotypes of P. aeruginosa. The MAbs also reacted with some of the other Pseudomonas spp., but not with P. cepacia or Xanthomonas (Pseudomonas) maltophilia. Cross-reactivity with whole cells from other bacterial species was minimal or not observed. Reactivity of MAbs with some Staph. aureus strains was observed, and binding to the protein A component was implicated. The reactivity of the MAbs was investigated further by flow cytometry and immunogold electronmicroscopy. The suitability of the MAbs for an immunological assay for detection of P. aeruginosa in respiratory secretions from CF patients is discussed.

Animals↗
PubMed1991

Monoclonal antibodies as probes for detecting lipopolysaccharide expression on Escherichia coli from different growth conditions.

Monoclonal antibody (mAb) probes were used to investigate the expression of lipopolysaccharide (LPS) on four Escherichia coli strains, grown under a variety of conditions in batch culture which mimicked some of the in vivo environmental conditions of an infected host. Techniques of silver staining, immunoblotting, whole cell ELISA and flow cytometry were all used to monitor the expression of LPS on the bacteria and the binding of the anti-LPS mAbs. Growth in heat-inactivated sheep serum and magnesium-depleted conditions demonstrated increased expression of LPS core and subsequent increased binding of anti-core mAbs. Magnesium-depleted conditions also resulted in decreased production of O-polysaccharide material. Iron-depleted bacteria showed only minor changes in LPS expression, although increased binding of anti-core mAbs was observed. Nitrogen-deficient/high-carbon conditions, chosen to promote capsule production, resulted in increased expression of O-polysaccharide and decreased binding of anti-core mAbs.

Antibodies, Monoclonal↗
PubMed1991

Use of rapid carbohydrate utilisation test for identifying "Streptococcus milleri group".

A short series of biochemical and serological tests were developed for the rapid presumptive identification of "Streptococcus milleri group" isolates. One hundred and seventy seven streptococcal isolates were recovered from the mouths of 10 out of 12 healthy adult volunteers by use of a simple sampling procedure and a single selective medium. In all, 127 oral "S milleri group" isolates were identified by biochemical and serological tests, confirming the indigenous nature of these streptococci in the mouth. Most (70.1%) of "S milleri group" isolates were non-haemolytic, 26% were alpha-haemolytic, and 3.9% beta-haemolytic. Fifty four (42.5%) were serologically typable, of which 46 were Lancefield group F, suggesting that the mouth is an important source of Lancefield group F streptococci. A collection of group F streptococci from a range of sources was indistinguishable from a collection of oral "S milleri group" isolates on the basis of the tests used, supporting the general synonymity of group F streptococcus with the broader "S milleri group". The battery of tests was cheap and simple to perform, and was capable of identifying "S milleri group" isolates from a range of sources, including variants with wide sugar fermentation patterns.

Adult↗
PubMed1990

A comparison of immunoblotting, flow cytometry and ELISA to monitor the binding of anti-lipopolysaccharide monoclonal antibodies.

This study was designed to assess the use of flow cytometry to observe the binding, under physiological conditions, of anti-lipopolysaccharide (LPS) monoclonal antibodies (mAbs) to whole bacteria, and to compare this with the more conventional whole cell ELISA and immunoblotting techniques. The bacteria consisted of two clinical isolates of E. coli 018:K1 and 06:K5 and two isogenic mutants of the 018 parent: a non-capsulate (018:K-) and a rough mutant (018rf). Two cross-reactive anti-core mAbs and one 018 0-antigen-specific mAb were used. ELISA and flow cytometry showed that capsule and O-polysaccharide influenced the binding of mAbs to the bacteria, whilst the latter technique demonstrated that sub-populations existed. Immunoblotting showed the two anti-core mAbs to be different, one bound only to core which was not substituted with O-antigen, whilst the other bound both to substituted and unsubstituted core. This comparison for monitoring the binding of anti-LPS mAbs demonstrates the potential use of flow cytometry in bacterial cell surface research, and complements results obtained by ELISA and immunoblotting.

Animals↗
PubMed1990

Characterisation of Bacteroides from sheep periodontal disease by SDS-PAGE of outer membrane proteins.

Isolates of Bacteroides species obtained from a longitudinal study of developing periodontal disease in sheep were analysed by SDS-PAGE. Protein profiles of Sarkosyl-insoluble outer membrane extracts were compared within groups of isolates which had already been defined by conventional biochemical techniques. Heterogeneity was exhibited within most groups. Isolates of B. gingivalis and B. asaccharolyticus shown to be similar to human isolates by conventional biochemical tests, gave different protein profiles from the respective type cultures. The sheep B. gingivalis-like isolates were however homogeneous, while the B. asaccharolyticus-like organisms could be divided into 3 subgroups. SDS-PAGE appears to be a useful tool for the examination of bacterial flora and recognition of subgroups of subspecies.

Animals↗
PubMed1990

A longitudinal study of the cultivable subgingival anaerobic bacteria isolated from sheep during the development of broken mouth periodontitis.

In a longitudinal bacteriological study of the cultivable subgingival anaerobic flora isolated from developing broken mouth periodontitis in sheep, samples were taken from five sheep on each of three farms on seven occasions over a period of 2.5 years. Ten different bacterial genera were isolated regularly but with fluctuating frequencies. Bacteroides and Fusobacterium organisms accounted for nearly 70% of the isolates. The Bacteroides and Fusobacterium isolates studied in detail from one farm were identified to species level. The fusobacteria comprised F. nucleatum-like organisms (68.6%). F. necrophorum (29.6%) and F. naviforme (1.8%). The Bacteroides spp. were divided into 11 main groups and included black-pigmented species similar to B. asaccharolyticus and B. gingivalis. On the farm studied in detail, the sheep could be allocated to two groups according to progression of periodontal disease. Most of the B. gingivalis-like isolates were from sheep with actively progressing disease, indicating that this organism may play a role in periodontal destruction in sheep.

Animals↗
PubMed1990

Outer membrane proteins of bovine strains of Pasteurella multocida type A and their doubtful role as protective antigens.

Outer membranes were prepared by the Sarkosyl method from 30 strains of Pasteurella multocida and the closely related Taxon 13, which had been isolated from cattle. The patterns of the outer membrane proteins (OMPs) on SDS-PAGE were generally similar to one another, though the four major proteins (a-d) varied somewhat in molecular mass; these patterns allowed the strains to be arranged into 12 groups. Taxon 13 strains and typical P. multocida strains were indistinguishable, both types being found within the same group. Mice were vaccinated with heat-killed bacteria of three strains and challenged with 10 LD50 of homologous and heterologous live bacteria, representing groups based on OMP patterns; the best protection was afforded by strain W674, which protected against nine of the 17 challenge strains; but there was no correlation between protection and PAGE pattern. Pre-vaccination and pre-challenge sera were used in immunoblotting to probe OMPs from protective and non-protective strains. All three vaccines produced antibody to proteins a and d; these proteins appeared to be common to all strains, varying in molecular mass but not in overall antigenic expression. The antibody response to the other two major OMPs appeared to be PAGE-group specific. There was no correlation between protection and the antigen pattern seen by immunoblotting.

Animals↗
PubMed1990

Production of mouse monoclonal antibodies to Pasteurella multocida type A and the immunological properties of a protective anti-lipopolysaccharide antibody.

Eight monoclonal antibodies (MAbs) were produced from mice immunised with whole cells of heat-killed Pasteurella multocida type A which had been cultured under iron-restricted conditions. The MAbs were selected by an enzyme-linked immunosorbent assay (ELISA) in which the antigen consisted of whole bacteria of the immunising strain. Their reactivity was investigated further by immunoblotting, indirect haemagglutination, a complement-mediated bactericidal assay and passive protection of mice. One of the eight MAbs was shown by immunoblotting to react with lipopolysaccharide (LPS), was bactericidal, and completely protected mice against homologous challenge with 10 LD50 of live bacteria. This MAb was selected for further study. Its reaction with LPS of 17 type-A strains and of single strains of types B, D and E was investigated by immunoblotting. Strains that reacted with the anti-LPS MAb in immunoblots were susceptible to its bactericidal activity and gave high ELISA absorbances. Those that did not react were not susceptible to its bactericidal activity and gave low ELISA readings. The relation between bactericidal activity and ELISA absorbance was highly significant (p less than 0.001). Five of the strongly reacting heterologous strains and one non-reacting strain were selected as challenge organisms in a passive protection experiment: only the mice receiving the reacting strains were protected.

Animals↗
PubMed1990

The susceptibility of in vivo-grown Pasteurella haemolytica to ovine defence mechanisms in vitro.

Pasteurella haemolytica organisms grown in vivo were examined for their susceptibility to ovine immune mechanisms in vitro. Compared with in vitro grown organisms they were less susceptible to opsonophagocytosis and, in contrast, susceptible to complement-dependent killing in the absence of exogenous antibody. These differences were not associated with phenotypic changes in the surface of the bacterial cell. However, overproduction and de novo synthesis of proteins was observed in in vivo grown organisms. Also, bound host-immunoglobulin was observed on in vivo grown organisms and a role for this in modifying the interaction with immune mechanisms is discussed.

Animals↗
PubMed1989

Preliminary study of the anaerobic bacteria isolated from subgingival plaque from sheep.

A preliminary study was made to determine the genera of cultivable anaerobic bacteria which could be isolated from subgingival plaque of sheep. Samples were taken from 10 sheep on farms with a known record of broken mouth periodontitis. For assessment of the sampling technique, samples were also taken from freshly exposed tooth roots in killed sheep. The bacteria isolated on several selective and non-selective media were identified to genus level by a combination of Gram reaction, colony morphology and gas chromatographic analysis of volatile and non-volatile fatty acid metabolic end products. At least 10 different genera were isolated and these findings are discussed in relation to the bacteriology of human periodontitis and recent studies of sheep broken mouth periodontitis.

Animals↗
PubMed1989

Immunochemistry of the cell surfaces of Bacteroides bivius and Bacteroides disiens.

Outer membranes were extracted from seven strains of Bacteroides bivius and six strains of B. disiens by the Sarkosyl method. Lipopolysaccharides (LPS) were extracted from the same strains by the Proteinase K method, and from three strains of each species by an aqueous phenol method. Analysis of the outer-membrane proteins by SDS-PAGE demonstrated that, within a species, very similar patterns with many shared or common bands were produced, but there were sufficient differences between species to allow separation. Immunoblotting with antisera raised against whole cells of each of the type strains showed that many antigens were shared between species. Smooth LPS was present in both species. By immunoblotting, the O-antigen of B. disiens was shown to be common to all six strains, and there was no cross-reaction between the B. disiens antiserum and B. bivius LPS. The O-antigen of B. bivius was not detected by immunoblotting with homologous antiserum, but antiserum to B. bivius reacted with a series of common low molecular mass antigens that were present in LPS preparations from strains of both species.

Bacterial Outer Membrane Proteins↗
PubMed1989

Typing of Clostridium difficile causing diarrhoea in an orthopaedic ward.

In an outbreak of diarrhoeal disease in an orthopaedic ward Clostridium difficile was isolated from all six patients with diarrhoea. Attempts were made to type these isolates by means of antibiogram, detection of pre-formed enzymes, analysis of surface proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting, and plasmid profile analysis. This showed that a single strain (type E) indistinguishable by the four distinct methods of typing, was isolated from all six patients at some time during their episodes of diarrhoea. Relapse was caused by the acquisition of a new strain in two patients, and by re-emergence or reacquisition of the original strain in two patients. The immunochemical method was the most sensitive and discriminatory of the typing strategies adopted.

Aged↗
PubMed1989

Changes in anti-endotoxin-IgG antibody and endotoxaemia in three cases of gram-negative septic shock.

Circulating endotoxin levels and IgG antibodies to a range of Gram-negative bacterial lipopolysaccharides (LPS) (endotoxins) of different sizes and structures were measured daily in three cases of septic shock. There was an inverse relationship between endotoxin levels and cross-reactive antibodies to the core glycolipid (CGL) region of lipopolysaccharide. This suggests that antibody to LPS-CGL was initially consumed by a superabundance of endotoxin, and that a resurgence of intrinsic anti-LPS-CGL antibody levels may be associated with a reduction of circulating endotoxin. The implications of these findings for passive antibody therapy of septic shock are discussed.

Adult↗
PubMed1986

Immunochemistry of the surface carbohydrate antigens of Bacteroides fragilis and definition of a common antigen.

The components extracted by aqueous phenol from whole cells of Bacteroides fragilis were analysed by SDS-PAGE and immunoblotting and shown to consist of a series of strain-specific, cross-reactive and common antigens. Regularly-spaced ladder patterns on silver-stained gels indicated that in most strains the LPS was present as a predominantly smooth type, but with chain lengths of varying molecular mass, ranging within each particular strain from essentially rough forms to long chain-length smooth forms. The rough form of the LPS at the gel front possessed an antigen common to most of the strains investigated. Another antigen, which migrated behind the rough LPS on SDS gels, was common to all strains of the species. The smooth LPS forms and the other high molecular mass components were strain-specific antigens. Previously published methods are not capable of producing pure LPS or capsular polysaccharide for this organism.

Antigens, Bacterial↗