PubMed Health⌕ Search

Biomedical subjects

T J Mitchell

Publications and source records attributed to T J Mitchell.

At least 109 records · Page 6Linked to original sources
PubMed1987

Biological mode of action of Clostridium difficile toxin A: a novel enterotoxin.

Antibody neutralisation and toxin A elution experiments showed that toxin A uptake from rabbit intestinal lumen was a continuous process. The kinetics of the ileal and colonic responses were significantly different; a much longer incubation (4 h) with toxin was required for colon, compared with 45 min for the ileum, to induce fluid accumulation at 12 h. Fluid secretion was induced only when toxin had gained access to deeper tissues, probably achieved by several toxin uptake-tissue damage cycles. Toxin A induced haemorrhage in both ileal and colonic tissues. In ileum, the villus architecture was severely damaged and this gave rise to protein-rich bloody luminal fluid. In the colon, although colonocytes were removed, the basement membrane remained intact; this resulted in a tissue-localised haemorrhage and a protein-low watery ultrafiltered luminal fluid. Toxin A is thus a novel type of histotoxic enterotoxin.

Animals↗
PubMed1987

The effects of Clostridium difficile crude toxins and toxin A on ileal and colonic loops in immune and non-immune rabbits.

Rabbits were solidly immunised by parenteral injection of purified Clostridium difficile toxin A such that they resisted an intravenous challenge with a normally lethal dose of toxin A. Ileal and colonic loops constructed in non-immune and immune animals received challenge injections of crude culture filtrate or purified toxin A of C. difficile. Protection of ileum was manifest after sufficient initial mucosal damage resulted in release of high levels of antitoxin A into the loop lumen of immune animals. There was less fluid accumulation in ligated ileal loops of immune than of non-immune rabbits. Less protection was observed when loops were challenged with crude culture filtrate containing toxins A and B than when challenged with purified toxin A. In-vitro studies with Ussing chambers yielded no evidence for tissue-localised immunity as judged by electrical responses and histology of toxin-treated tissue from non-immune and immune animals. No differences were found in the degree of epithelial damage, or volume or composition of fluid accumulating in colonic loops of non-immune and immune rabbits challenged with toxin A or crude culture filtrate. However, in colonic loops of immune rabbits there was no overt tissue-localised haemorrhage, whereas in those of non-immune rabbits tissue-localised haemorrhage was marked. In contrast to our findings with ileal loops, fluid accumulating in colonic loops was watery and contained substantially less total protein and (in immune animals) antitoxin A.

Animals↗
PubMed1987

Clostridium difficile--a spectrum of virulence and analysis of putative virulence determinants in the hamster model of antibiotic-associated colitis.

Each of nine different toxigenic strains of Clostridium difficile was administered orally to groups of hamsters pre-treated with clindamycin and housed individually in sterile isolator boxes. Faecal pellets and caecal contents from well, diarrhoeic, moribund and freshly dead animals were analysed for C. difficile and toxins A (enterotoxin) and B (cytotoxin), and tissue obtained when animals were killed was examined histologically. Not all strains were equally virulent in this model. Four strains of C. difficile killed all animals within 48 h and are designated as highly virulent for hamsters. These strains were clinical isolates from three cases of disease in man and one case in a hamster. Five strains caused death of some animals but only after 5 and upt to 13 days and are designated as less virulent for hamsters. These strains were isolated from asymptomatic infants (2) and household pets (2), and from the environment (1). The surviving test hamsters were killed after 14 days and, in most cases, were colonised by C. difficile, though levels of toxins A and B in caecal contents were low. None of the cultures used for challenge was capsulate or hydrophobic. There was no correlation between virulence and production of toxins A and B in vitro in tryptic-nitrate broth. With two strains examined, there was a correlation between virulence and toxin A (but not toxin B) production in caecal emulsions derived from clindamycin pre-treated hamsters. Caecal contents from the majority of moribund and freshly dead animals had quantities of toxin A sufficient to cause disease or death if given orogastrically. Toxin B was not produced in a fixed ratio with toxin A. The data support the view that high virulence of C. difficile is determined by efficient disease-inducing colonisation of the gut and the ability to generate, rapidly, high levels of toxin A in vivo.

Animals↗
PubMed1986

Growth of Clostridium difficile and production of toxins A and B in complex and defined media.

The ability of several strains of Clostridium difficile to grow and to produce toxins A and B in complex and defined culture media has been studied with special reference to the amino-acid composition of the medium. The production of these toxins varied with the strain used and with the composition of the growth medium. Toxin A production was not inextricably linked to production of toxin B since conditions were found in which only one or other toxin was produced.

Amino Acids↗
PubMed1986

Sporogenesis and toxin A production by Clostridium difficile.

The kinetics of spore production by Clostridium difficile were not paralleled by release of C. difficile toxin A in vitro. Toxin A was not found to be associated with either purified whole spores or spore coats. Residual traces of toxin A detected in spore contents were almost certainly derived from contaminating vegetative cell debris. Thus, toxin A is unlikely to be a spore constituent or associated with sporogenesis.

Bacterial Toxins↗
PubMed1986

Effect of toxin A and B of Clostridium difficile on rabbit ileum and colon.

The effect of purified toxin A and partially purified toxin B on rabbit ileum and colon was investigated. Toxin A caused tissue damage which was followed by permeability changes and fluid accumulation in both tissues. Toxin A did not increase the permeability of the colon to the extent observed for ileum; secreted fluid contained less protein of plasma origin. Toxin B had no effect on either tissue. Secretory and tissue damaging properties of crude C difficile toxins were found to be due to toxin A.

Animals↗
PubMed1985

Stimulator requirements for primed alloreactive T cells: macrophages and dendritic cells activate T cells across all genetic disparities.

The cellular requirements for stimulating primed alloreactive T cells have been investigated. In vitro-primed secondary alloreactive cells, long-term lines, and Ly 1+2- noncytolytic clones which reacted with allo-H-2K, D, or Mls (M locus) antigens were tested. The data indicated that a specialized antigen-presenting cell such as a macrophage or a dendritic cell was required for stimulating primed alloreactive cells across all the genetic disparities tested. B and T lymphocytes were ineffective stimulators. The stimulator requirement for secondary and Ly 1+2- clone responses was heterogeneous, since both macrophages and dendritic cells were effective stimulators. Thus, the allostimulator requirement for inducing proliferation and mediator secretion by the primed T-cell populations closely paralleled the requirement for stimulating unprimed populations. The only exception found was the peritoneal washout population, which did not stimulate a primary response but did stimulate secondary responses. The failure of peritoneal macrophages to stimulate a primary response was shown to be due to an inhibitory pathway which did not occur when the responding population was alloantigen primed.

Animals↗
PubMed1985

Antigen presentation by spleen dendritic cells.

It is now recognized that dendritic cells (DC) isolated from mouse spleen play an important role in activating T lymphocytes. These DC, which show many similarities to veiled cells found in the paracortical regions of spleen and lymph node, may be closely related to the epidermal Langerhans cells. It is known that DC are extremely effective allostimulators. We have also found that although DC lack demonstrable phagocytic ability, they are extremely potent at presenting soluble polypeptide antigens to primed T cells. Since T lymphocytes comprise several distinct subsets (particularly cytotoxic, helper, and suppressor) in our most recent studies we have asked whether DC are able to trigger all these different subsets of T cells. We examined the ability of different spleen cell types coupled with the hapten NP to induce antigen-specific T suppressors for a delayed type hypersensitivity (DTH) response. It was found that T suppressors were generated only when hapten was conjugated to a spleen-derived antigen-presenting cell. Further analysis revealed that macrophages but not DC were able to induce defined sets of suppressor cells in vivo (although DC were able to trigger a very powerful DTH response). We also examined the ability of DC to activate T cells which are required to cooperate with B cells in the production of antibody. Even though DC were able to trigger T lymphocytes to produce lymphokines, these activated T cells did not act as helper cells in a standard hapten-carrier system. Possible mechanisms for this dichotomy of DC function are discussed.

Animals↗
PubMed1984

Production and release of toxins A and B by Clostridium difficile.

The production and release of toxins A and B by Clostridium difficile during in-vitro culture was investigated. Cell-associated toxin A was detected by immunoelectrophoresis of bacterial extracts released by ultrasonication and by fluorescent antibody labelling of whole cells. Extracellular toxin A was detected by immunoelectrophoresis and by enzyme-linked immunosorbent assay; extracellular toxin B was detected by cytotoxin assay. Both toxins A and B were produced and released during the decline phase of the bacterial growth cycle. The possible significance of these results in relation to the pathogenesis of pseudomembranous colitis is discussed.

Bacterial Proteins↗
PubMed1984

Limiting values for the RBE of fission neutrons at low doses for life shortening in mice.

We have analyzed recently published data on the effects of low doses of fission neutrons on the mean survival times of mice. The analysis for single-dose exposures was confined to doses of 20 rad or less, while for fractionated exposures only total doses of 80 rad or less were considered. We fitted the data to the frequently used power function model: life shortening = beta D lambda, where D is the radiation dose. We show that, at low doses per fraction, either (1) the effects are not additive or (2) the dose-effect curve for single exposures cannot show a greater negative curvature than about the 0.9 power of dose. Analysis of the data for gamma rays showed that an exponent of 1.0 gave an acceptable fit. Taken together, these findings indicate that the RBE for neutrons cannot change more rapidly with neutron dose than about RBEN approximately = k/D0.1N. This conflicts with the more widely accepted relationship, RBEN approximately = k/D0.5N. Because of the inherent implausibility of exponents less than 1.0 for the neutron dose-effect curves at low doses we conclude that at neutron doses of 20 rad or less the RBE for life shortening is constant and ranges from 13 to 22 depending on mouse strain and sex.

Animals↗
PubMed1984

Nonparametric estimation of the distribution of time to onset for specific diseases in survival/sacrifice experiments.

This paper concerns the analysis of an animal survival/sacrifice experiment designed to investigate the incidence of a particular disease of interest. The disease is assumed to be irreversible, and detectable only at death, for example by a necropsy. Each observation can be of one of three types: (i) death caused by the disease, (ii) death from a competing cause such as sacrifice, with the disease present, or (iii) death with the disease absent. A two-dimensional EM algorithm is proposed for the nonparametric maximum likelihood estimation of the distributions of the time to onset and of the time to death from the disease. These can be compared with nonparametric estimators recently proposed by Kodell , Shaw and Johnson (1982, Biometrics 38, 43-58) and by Dinse and Lagakos (1982, Biometrics 38, 921-932). A slight modification of the algorithm permits the construction of likelihood-based interval estimates of quantiles of the distributions. Some extensions and generalizations are indicated.

Age Factors↗
PubMed1979

Log-linear models in the analysis of disease prevalence data from survival/sacrifice experiments.

This paper considers the problem of analyzing disease prevalence data from survival experiments in which there may also be some serial sacrifice. The assumptions needed for "standard" analyses are reviewed in the context of a general model recently proposed by the authors. This model is then reparametrized in log-linear form, and a generalized EM algorithm is utilized to obtain maximum likelihood estimates of the parameters for a broad class of unsaturated models. Tests based on the relative likelihood are proposed to investigate the effects of treatment, time, and the presence of other diseases on the prevalences and lethalities of specific diseases of interest. An example is given, using data from a large experiment to investigate the effects of low-level radiation on laboratory mice. Finally, some possible directions for future research are indicated.

Animals↗
PubMed1978

Exploratory analysis of disease prevalence data from survival/sacrifice experiments.

The paper considers the problem of analyzing disease prevalence data from survival experiments in which there may also be some serial sacrifice. The primary objective of the analysis is to describe the composition of the treated and control populations, in terms of age-dependent disease prevalences, by removing distortions in the data caused by the biased nature of the primary sampling mechanism (death). The statistical model which is utilized for this purpose is parameterized in terms of illness state prevalences and lethalities. It does not require determination of cause of death; nor does it assume that diseases progress independently. Methods are presented for estimating various quantities of interest, including disease-specific relative risks and measures of association among diseases. An application of this analysis is shown, using data from a large experiment to investigate the effects of low-level radiation on laboratory mice.

Age Factors↗