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Biomedical subjects

N J Gilmour

Publications and source records attributed to N J Gilmour.

At least 19 recordsLinked to original sources

p-Phenylenediamine allergy: the role of Bandrowski's base.

p-Phenylenediamine (PPD) is a commonly used hair-dye and a potent skin allergen. The mechanism of sensitization is unknown, as PPD is protein unreactive. We studied Bandrowski's base (BB), a PPD trimer, as well as 1,4-benzoquinone (BQ), a PPD hapten. PPD patch-test positive patients were patch-tested to BB and BQ. All tests were negative to 0.01% BQ and 0.01% BB. Five of 14 (35.7%) tested had true positive reactions to 0.1% BQ. One percent BQ was found to be irritant. Seven of 43 tested (16%) were positive to either 0.1% or 1% BB. The positive reactions to BB were weak, even when PPD reactions were strong. Mice lymph node assay gave EC3 values of 0.14% for PPD compared with 0.03% for BB. Therefore, BB is approximately 10 times more potent than PPD, taking into account the molarity. We suggest that while PPD may act as a prohapten, there is probably a spectrum of antigenic determinants in vivo. BB may be bound or metabolized by keratinocytes before it reacts with Langerhans cells.

Animals↗

Utility of historical vehicle-control data in the interpretation of the local lymph node assay.

The accepted approach to the interpretation of local lymph node assay (LLNA) data requires comparison of responses in the test groups with background activity found in concurrent vehicle-treated controls. However, of established value in the interpretation of toxicity test data is the use of historical control values that provide one criterion against which to judge the integrity of individual experiments. Specifically, the availability of robust and relevant historical control data permits examination of whether, in any individual experiment, control values fall within the expected range. With the most commonly used vehicle employed in the LLNA, acetone/olive oil (4 : 1) (v/v), the mean values, standard deviations and normal ranges are increasingly well established for a given laboratory, although there is some variation between laboratories, particularly with regard to expected ranges. Against this background, it is possible to identify (and, if appropriate, eliminate) a concurrent vehicle-control value that falls well outside the expected range. To explore critically the potential merits of this approach, one specific example is examined in detail.

Animals↗

Vaccine containing iron-regulated proteins of Pasteurella haemolytica A2 enhances protection against experimental pasteurellosis in lambs.

A vaccine containing sodium salicylate extract (SSE) of Pasteurella haemolytica A2 cells grown in a medium chemically depleted of available iron by the addition of alpha alpha dipyridyl to induce iron-regulated proteins (IRPs) conferred protection to specific pathogen-free (SPF) lambs exposed to an aerosol of P. haemolytica A2. The disease score in these lambs was significantly lower (p less than 0.005) than those in unvaccinated lambs or in lambs immunized with SSE prepared from cells grown in iron-replete medium. Immunoblotting of sera from these SPF lambs against whole cell antigens of P. haemolytica A2 grown under iron-restricted conditions demonstrated that antibodies to IRPs were present only in the sera of animals immunized with SSE-IRP. The antibody profile of sera from the SSE-IRP group was similar to that obtained with serum from a lamb which had recovered from P. haemolytica A2 disease produced experimentally. Negligible levels of cytotoxin-neutralizing and bactericidal antibodies were detectable in the SSE-IRP group and therefore appear not to be involved in the protection observed in this experiment.

Animals↗

[Pasteurella haemolytica serotyping].

Forty-nine strains of Pasteurella haemolytica were serotyped in a pilot study. The majority of strains were isolated from sheep which had died from fibrinous pneumonia and small numbers from cases of sepsis, polyserositis, mastitis and meningitis. Serotype A2 was found to be the most prevalent: 69.4 per cent. In addition, serotype A7 with 18.4 per cent and serotypes A1 and A6, both 6.1 per cent, were identified. Biotype T strains were not detected in this study. Further research is required into the seasonal prevalence of P. haemolytica serotypes in sheep and cattle. This knowledge may help to improve herd health schemes for sheep and cattle in the Netherlands.

Hemagglutination Tests↗

Use of long-acting oxytetracycline against pasteurellosis in lambs.

The efficacy of long-acting oxytetracycline in the control of pneumonic pasteurellosis in lambs was tested on seven Scottish farms. After laboratory confirmation of pasteurella-related deaths in lambs, half the lambs in each flock were given long-acting oxytetracycline (20 mg/kg intramuscularly) and half were left untreated. On three farms a single treatment was given and on four farms two doses were administered four days apart. Eighteen of the 878 control lambs died as a result of confirmed Pasteurella haemolytica pneumonia compared with one of the 878 treated lambs. In addition nine of the control lambs were diagnosed clinically to have pasteurellosis which responded to treatment with oxytetracycline. None of the treated lambs were seen to be ill during the trial.

Animals↗

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↗

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↗

A plasmid which can be transferred between Escherichia coli and Pasteurella haemolytica by electroporation and conjugation.

Three broad-host-range plasmids (pRK290, pSa4 and pKT230) and one native Pasteurella haemolytica plasmid (pPH33) were used in transformation experiments with P. haemolytica strains T179 (serotype A1), Y216 (serotype A2) and its capsular-deficient variant Y216/NS1. No transformants were detected with either heat-shock or freeze-thaw techniques. However, by electroporation, all P. haemolytica strains were transformed by pPH33 but not by pRK290 or pSa4. The highest frequency obtained was 91 x 10(4) transformants per microgram of pPH33 DNA with P. haemolytica strain Y216/NS1. Although pPH33 itself was non-transmissible by conjugation, it could be mobilized from Escherichia coli, using the transfer function of the IncP plasmid pRK2013, into P. haemolytica at a frequency of 0.3-2.2 x 10(-3) per recipient cell.

Conjugation, Genetic↗

Characterisation of a new serotype of P haemolytica isolated in Hungary.

Three strains belonging to a new serotype of Pasteurella haemolytica (A16) were isolated from lambs and a wild boar in Hungary. The identity and validity of the new serotype was proved by biochemical tests and by the indirect haemagglutination test using unabsorbed and absorbed hyperimmune sera raised in rabbits.

Animals↗

Immunity of specific pathogen-free lambs to challenge with an aerosol of Pasteurella haemolytica biotype A serotype 2. Pulmonary antibody and cell responses to primary and secondary infections.

Specific pathogen-free (SPF) lambs previously exposed to an aerosol of P. haemolytica biotype A serotype 2 (A2) were immune to subsequent challenge with an aerosol of P. haemolytica A2. Untreated control lambs were not immune to this challenge. The local immune responses of the lung to these challenges were examined. High IgG and IgA titres to P. haemolytica and high levels of opsonizing antibody against P. haemolytica were present in the lung washings from previously infected immune lambs at autopsy, seven days after the second infection. Lung washings from control lambs, 7 days after challenge with P13 virus and P. haemolytica A2, had no IgG titres, very little opsonizing activity but did have IgA titres which were significantly higher than in unchallenged control lambs. The cellular response of animals challenged with P13 virus and P. haemolytica was significantly greater than that of unchallenged controls or of lambs exposed only to P. haemolytica. However, this finding was complicated by the response to P13 virus. Lymphocytes from lung washings of all lambs failed to respond in a lymphocyte stimulation test to phytohaemagglutinin while blood lymphocytes did respond. There was little specific response to P. haemolytica antigen in the test.

Animals↗