Biomedical subjects
M J Day
Publications and source records attributed to M J Day.
Cytokine mRNA expression in mucosal biopsies from German shepherd dogs with small intestinal enteropathies.
German shepherd dogs (GSD) are predisposed to enteropathies such as inflammatory bowel disease (IBD) and small intestinal bacterial overgrowth (SIBO). The present study examined the role of cytokines in the immunopathogenesis of both conditions. Duodenal mucosal biopsies were taken from GSDs with small intestinal enteropathies (group 1; N = 16) or control dogs (group 2, N = 12). IL-2, IL-4, IL-5, IL-10, IL-12p40, IFN-gamma, TNF-alpha, and TGF-beta1 mRNA expression was determined by semiquantitative reverse transcriptase polymerase chain reaction. IL-2, IL-5, IL-12p40, TNF-alpha, and TGF-beta1 mRNA expression in group 1 dogs was significantly greater than in group 2 dogs (all P<0.01), but there were no significant differences between dogs with IBD or SIBO. Further, antibiotic treatment in five dogs with SIBO, resulted in reduced TNF-alpha and TGF-beta1 mRNA expression (P<0.05). Such alterations in cytokine mRNA expression suggest heightened immune responses within the duodenal mucosa in GSDs with either SIBO or IBD.
UK consumption databases relevant to acute exposure assessment.
Consumption data can be expressed in different ways although it should be comparable to what is known about the toxicological property of the chemical in question and the likely pattern of consumer exposure. For example if the chemical has an acute toxic effect the consumption data should ideally be based on a single eating occasion or consumption over a single day and be based on the population of concern. This paper describes five UK dietary surveys and consumption data, specifically relevant to acute exposure assessment, from adult and pre-schoolchildren surveys. The acute consumption data are taken from the first day of the dietary regime. Closer examination of the dietary survey information suggests that either picking a random survey day or selecting all consuming days give 97.5 percentile values which are not significantly different. Consumption data are given for whole commodities and a few fruit juices. Examination of the dietary survey information shows that the consumption patterns are very different between whole commodities (e.g. apples), ingredients (e.g. apple in a dish or recipe) and juices and between adults and young children.
Epitheliotropic lymphoma (mycosis fungoides) presenting as blepharoconjunctivitis in an Irish setter.
An 11-year-old neutered female Irish setter was presented with a six-week history of blepharoconjunctivitis affecting the right eye. A conjunctival biopsy was taken and histopathological examination revealed a heavy cellular infiltrate involving the epithelial and subepithelial tissues. Immunohistochemical staining showed the intraepithelial cell population to uniformly have the phenotype CD3 (T-cell specific marker). A diagnosis of epitheliotropic lymphoma (mycosis fungoides) was made. The use of a synthetic retinoid and topical prednisolone in the management of the case is discussed.
Intestinal haemangiosarcoma in the cat: clinical and pathological features of four cases.
The clinical and pathological features of four cases of feline intestinal haemangiosarcoma are described. All cases were in domestic shorthaired cats and the mean age of the animals (n=3) was 13 years. The tumours originated in the colon, small intestine, ileocaecocolic junction or rectum. The rectal tumour was juxtaposed with an adjacent mast cell neoplasm. Metastasis to mesenteric lymph node occurred in two cases, and in one of these cats there was also abdominal seeding. The histopathological appearance was of a spindle cell neoplasm with vascular differentiation in each case. Immunohistochemical staining for factor VIII-related antigen, an endothelial cell marker, confirmed all four tumours to be of endothelial origin. The neoplastic endothelial cells lining irregular vascular channels were more likely to express the antigen than those forming denser sheets without obvious vascular differentiation.
Pemphigus foliaceus in association with systemic lupus erythematosus, and subsequent lymphoma in a cocker spaniel.
A seven-year-old neutered male cocker spaniel was presented with an 11-month history of generalised bacterial dermatitis. There were skin lesions over the entire body, which were round, slightly raised and encrusted. Skin biopsies were collected and the histological findings were consistent with pemphigus foliaceus. Immunohistochemical staining by the indirect immunoperoxidase method was positive, with desmosomal deposition of immunoglobulin (Ig) G. Haematological analysis revealed a regenerative anaemia and profound thrombocytopenia, while a Coombs' test was positive for polyvalent canine Coombs' reagent and anti-dog IgG. An antinuclear antibody test was positive, with a titre of 10,240. An ophthalmic examination demonstrated low tear production (keratoconjunctivitis sicca). Seven months after initial referral, the dog was re-presented with severe generalised peripheral lymphadenopathy. Radiographic evaluation of the thorax and abdomen revealed enlarged cranial mediastinal and sublumbar lymph nodes. Tru-Cut biopsy from an enlarged lymph node confirmed the diagnosis of lymphoma, which was phenotyped as a B-cell tumour. The diagnosis in this case was systemic lupus erythematosus, with the unusual feature of pemphigus foliaceus, and subsequent development of B-cell lymphoma. The case adds further to knowledge of the protean clinical presentations of canine autoimmune diseases and provides additional evidence for the potential association between autoimmunity and immune-system neoplasia in this species.
Juvenile nephropathy in a boxer, a rottweiler, a collie and an Irish wolfhound.
Juvenile nephropathy was diagnosed in a Boxer, a Rottweiler, a Collie and an Irish Wolfhound dog, each presenting with signs compatible with chronic renal failure. The diagnosis in each case was based on the presence of persistence of poorly differentiated tissue (immature glomeruli and/or tubules, persistent mesenchyme) on histopathologic examination. Although juvenile nephropathy has been reported in many breeds of dog, this is the first report of the condition in the Collie and the Irish Wolfhound and only the second description in the Boxer and the Rottweiler. The possibility of an inherited origin of the condition in these four breeds is at present unknown.
Seasonal population dynamics and interactions of competing bacteriophages and their host in the rhizosphere.
We describe two prolonged bacteriophage blooms within sugar beet rhizospheres ensuing from an artificial increase in numbers of an indigenous soil bacterium. Further, we provide evidence of in situ competition between these phages. This is the first in situ demonstration of such microbial interactions in soil. To achieve this, sugar beet seeds were inoculated with Serratia liquefaciens CP6RS or its lysogen, CP6RS-ly-phi 1. These were sown, along with uninoculated seeds, in 36 field plots arranged in a randomized Latin square. The plots were then sampled regularly over 194 days, and the plants were assayed for the released bacteria and any infectious phages. Both the lysogen and nonlysogen forms of CP6RS survived equally well in situ, contradicting earlier work suggesting lysogens have a competitive disadvantage in nature. A Podoviridae phage, identified as phi CP6-4, flourished on the nonlysogen-inoculated plants in contrast to those plants inoculated with the lysogen. Conversely, the Siphoviridae phage phi CP6-1 (used to construct the released lysogen) was isolated abundantly from the lysogen-treated plants but almost never on the nonlysogen-inoculated plants. The uninoculated plants also harbored some phi CP6-1 phage up to day 137, yet hardly any phi CP6-4 phages were found, and this was consistent with previous years. We show that the different temporal and spatial distributions of these two physiologically distinct phages can be explained by application of optimal foraging theory to phage ecology. This is the first time that such in situ evidence has been provided in support of this theoretical model.
Antigen specificity in canine autoimmune haemolytic anaemia.
Autoimmune haemolytic anaemia (AIHA) is the most common clinical manifestation of autoimmunity in the dog and generally presents as a profound, regenerative, Coombs' positive anaemia of acute or chronic onset. The disease pathogenesis involves formation of erythrocyte-specific autoantibodies of the IgG and IgM class that may fix complement resulting in intra- or extravascular haemolysis. Western blotting and immunoprecipitation studies using autoantibody eluted from the erythrocytes of dogs with AIHA have demonstrated specificity for erythrocyte glycophorins and the membrane anion-exchange molecule (band 3). Autoantibodies specific for the cytoskeletal molecule spectrin have been identified in serum by ELISA. The specificity of autoreactive T-cells has been examined in vitro using bulk cultures stimulated with a panel of autoantigens including intact erythrocyte membranes, purified glycophorin and spectrin fractions and a panel of overlapping 15-mer glycophorin peptides. Control responses to ConA and recall (vaccine antigens) and non-recall (KLH) antigens were measured in the same system. PBMC obtained from dogs that had recovered from AIHA consistently proliferated in response to erythrocyte membranes, with occasional responses to spectrin or glycophorin. PBMC from sone clinically normal dogs also responded to erythrocyte membranes. PBMC obtained from dogs closely related to AIHA cases gave the most consistent responses, including proliferation when stimulated by the glycophorin peptides. These data suggest that normal dogs harbour erythrocyte autoreactive lymphocytes, and that these cells may be primed in dogs recovered from AIHA or having genetic susceptibility to the disease.
Immunohistochemical study of the inflammatory infiltrate associated with feline cutaneous squamous cell carcinomas and precancerous lesions (actinic keratosis).
The distribution of T lymphocytes (CD3+), B lymphocytes (CD79+), immunoglobulin-containing plasma cells (IgG, IgM and IgA), macrophages (Mac387+) and MHC Class II antigen was analysed in the inflammatory infiltrate associated with cutaneous squamous cell carcinomas (SCC) from 23 cats. Peri-tumoural skin (12 cases) and precancerous lesions of actinic keratosis (nine cases) were also evaluated for the expression of MHC Class II. The results revealed that an abundant inflammatory infiltrate was associated with the majority of SCC. This infiltrate was composed mainly of CD3+ T lymphocytes, B cells (CD79+) and IgG-bearing plasma cells, and the intensity of infiltration increased with the degree of invasiveness of the tumour. The number of CD3+ T cells and CD79+ cells was significantly increased in well-differentiated SCC compared with moderately differentiated tumours, whereas the number of IgM+, IgA+ plasma cells and Mac387+ macrophages was low or moderate and did not change significantly with histologic grade or invasiveness. MHC Class II antigen was expressed by infiltrating lymphocytes and macrophages, and by fibroblasts. A variable number of neoplastic cells (10% to 80%) in 10 SCC, and keratinocytes of basal layers in seven of nine cases of actinic keratosis also expressed MHC Class II, whereas keratinocytes of normal skin were always negative for this antigen. These results suggest that CD3+ T lymphocytes, CD79+ B cells and IgG-bearing plasma cells may participate in down-regulation of tumour growth, since these cell types were particularly numerous in well-differentiated and mildly invasive SCC, as well as in actinic keratosis. The expression of MHC Class II by neoplastic cells could enhance this local anti-tumour immune response.
Structural changes induced by mupirocin in Staphylococcus aureus cells.
Cells of mupirocin-sensitive, moderately-resistant and highly-resistant cultures of Staphylococcus aureus (mupirocin MICs 0.13, 16 and > 512 mg/l, respectively) were exposed to various concentrations of the antibiotic and examined by transmission electron microscopy. The most severe damage occurred in mupirocin-sensitive cells. Cells from moderately-resistant cultures trained in vitro to high-level mupirocin resistance were more hydrophobic than the parent cells. The antibiotic was slowly lethal to the mupirocin-sensitive strain and sub-inhibitory concentrations prevented or reduced growth of the other strains over a 6 h incubation period, irrespective of whether the drug was added at zero time or in the exponential growth phase.
Clues for the development of food-based dietary guidelines: how are dietary targets being achieved by UK consumers?
Expert scientific advice to the UK Government has been translated into eight general dietary guidelines, which form the core of population-based dietary advice in the UK and are supplemented by a food selection guide showing the types and proportions of foods needed for a balanced and healthy diet. Data from the Dietary and Nutritional Survey of British Adults were used to identify statistically significant differences between subgroups of the study population that met, or failed to meet, population nutritional goals for intakes of total fat, saturated fat and dietary fibre. Several eating habits--including greater consumption of starchy foods (particularly wholemeal varieties), greater consumption of fruit and the substitution of reduced-fat milk for whole-fat milk--were shared by the subgroups that met each of the nutritional goals. This analysis provides clues for any future refinement of food-based dietary guidelines.
Mupirocin resistance in staphylococci: development and transfer of isoleucyl-tRNA synthetase-mediated resistance in vitro.
Mupirocin resistance could be transferred from highly resistant clinical isolates of Staphylococcus aureus to highly sensitive recipients of Staph. aureus, Staph. epidermidis and Staph. haemolyticus. Transconjugants of the latter two organisms could transfer this resistance into mupirocin-sensitive Staph. aureus. Moderately resistant strains did not transfer this resistance to sensitive recipients, nor did strains with high-level mupirocin resistance developed by serial transfer or habituation. The inhibitory effects of mupirocin on crude isoleucyl-tRNA synthetases (IRS) isolated from mupirocin-sensitive and -resistant strains of Staph. aureus have been determined. Drug concentrations needed to produce 50% inhibition, I50 values, were very low against IRS from a highly sensitive strain, somewhat higher against IRS from moderately resistant strains, much higher against enzyme from strains trained in vitro to high-level resistance, and considerably higher still against IRS extracted from clinical isolates possessing high-level mupirocin resistance and from the transconjugates of such strains resulting from crosses with mupirocin-sensitive strains. It is concluded that high-level resistance in clinical isolates is plasmid-mediated involving a second, mupirocin-resistant IRS whereas in moderately resistant strains, and in strains trained in vitro to high-level resistance, chromosomal mutations are likely to be responsible for decreasing IRS sensitivity.
Rapid evaluation of biocidal activity using a transposon-encoded catechol 2,3-dioxygenase from Pseudomonas putida.
Pseudomonas putida (UWC1), containing a genetically-engineered plasmid (pQM899), that encodes for the production of catechol 2,3-dioxygenase (C230), was used as a potential means of rapidly estimating bactericidal activity of chlorhexidine diacetate (CHA), phenol, cetylpyridinium chloride (CPC) and phenylmercuric nitrate (PMN). Enzyme C230 converts catechol to 2-hydroxymuconic semialdehyde (2-HMS), which is yellow in colour, via a meta cleavage pathway. Ideal conditions for production and measurement spectrophotometrically of 2-HMS were determined. However, the correlation between this method and viable plate counts was not sufficiently accurate to enable 2-HMS production to provide a sufficiently sensitive determination of biocidal activity. An alternative method, synchronous scanning fluorimetry, in which the decrease in catechol concentration was measured under standardized conditions, provided a good dose-response histogram for all the biocides tested. Although, in comparison with plate counts, there was an underestimation of the bactericidal effects of phenol an PMN, the results of this study suggest that this method has potential in determining the bactericidal efficacy of agents such as CHA and CPC.
B-cell (CD79a+) lymphoma affecting the tarsal joint synovia in a sheep.
A lymphoma affecting the synovia was identified in both tarsal joints of an aged sheep. With a panel of cross-reactive antibodies specific for lambda and kappa immunoglobulin light chain, MHC class II, CD3, CD79a and the Ki-67 antigen, the neoplasm was classified as a B-cell lymphoma with uniform expression of surface membrane CD79a. This would seem to be the first report of a lymphoma affecting the synovia in a domestic animal, and the first use of the CD79a reagent on ovine tissue.
Analysis of leucocyte subsets in the canine intestine.
Immunohistochemistry and computer-aided morphometric analysis were used to define populations of leucocyte subsets in the intestinal tract of an outbred population of dogs with no evidence of gastrointestinal disease. In the small intestinal lamina propria, B cells and plasma cells (IgA+, IgM+ and IgG+) were prominent in peri-crypt regions, with a significant trend for a reduction in the number of cells towards the villous tip (P < 0.0001). By contrast, lamina propria T cells (CD3+) and T-cell subsets (CD4+ and CD8+) were present in greatest numbers at the tip of the villus, with significantly decreasing numbers towards the crypt regions (P < 0.0001). In the lamina propria, CD4+ cells outnumbered CD8+ cells (P = 0.05), but the opposite was true of the epithelial compartment (P < 0.001). The distribution of CD5+ lymphocytes was similar to that of CD3+ cells, in both the lamina propria and epithelial compartments. The numbers and distribution of cells expressing MHC class II, L1 and CD45 were recorded. Numerous eosinophils were present in the lamina propria, and an intra-epithelial population was also noted, especially in the crypt epithelium. Mast cells, which were mainly found in the subepithelial lamina propria, were also present within muscle layers, and cells expressing IgE had a similar distribution. Similar populations of cells were recorded in the colonic lamina propria and epithelium. The quantitative and qualitative data from this study will enable comparisons to be made with dogs suffering from inflammatory bowel diseases.
T-cell-rich B-cell lymphoma in the cat.
The clinical and pathological features of eight cases of feline T-cell-rich B-cell lymphoma are described. The disease occurred in older cats (mean age 11.4 +/- 3.9 years), which on initial examination generally showed enlargement of a single submandibular or cervical lymph node. After excision, there was no recurrence of the lesions at 6 months in three cats. In one further case, however, the lesion had recurred 6 months later; it was again excised but recurred after an additional 6 months. Microscopically, there was effacement of normal lymph node architecture by a nodular (n = 4) or diffuse (n = 4) proliferation of small to blastic lymphocytes, accompanied by a characteristic population of bizarre giant, or multinucleate, cells. The mitotic rate was low and mitoses were restricted to the atypical population. Immunophenotyping revealed the smaller lymphocytes to be a mixture of CD3+ MHC Class II+ T lymphocytes and BLA36+CD79variable MHC Class IIvariable B lymphocytes. The atypical cells were of the B-cell lineage (BLA36+MHC Class IIvariable). Polymerase chain reaction analysis revealed no proviral DNA products of feline leukaemia virus or feline immunodeficiency virus in tissue from any tumour, confirming that these neoplasms were not associated with either virus. The clinical, histological and immunophenotypic findings in these cats were identical with those of "nodular lymphocyte predominance (lymphocytic and histiocytic/L&H) Hodgkin's disease" in man.
Leishmaniosis and generalized demodicosis in three dogs: a clinicopathological and immunohistochemical study.
This paper describes the clinicopathological and immunohistochemical aspects of the skin lesions in three dogs with leishmaniosis and generalized demodicosis. Diffuse alopecia, crusts, folliculitis and furunculosis, as commonly seen in generalized demodicosis, were prominent in all the dogs. MicroIscopically, there was a diffuse and perifollicular superficial and deep granulomatous dermatitis and, in two dogs, both Copyright Demodex canis mites and Leishmania spp. amastigotes were observed in the same lesions. Numerous Mac387(+)macrophages were observed in the inflammatory infiltrates, but macrophages loaded with amastigotes were Mac387(-). In all cases, immunoreactive CD3 lymphocytes were sparse, both in the granulomatous and perifollicular infiltrates. There were numerous IgG+, IgG4(+)-secreting plasma cells in areas of folliculitis and furunculosis and fewer IgG2(+), IgG3(+), IgA+and IgM+-secreting plasma cells in the inflammatory infiltrate. In all cases, MHC Class II was expressed by the majority of dermal macrophages and dendritic cells, as well as by lymphocytes and fibroblasts. The paucity of CD3(+)lymphocytes, usually abundant in D. canis lesions, points to leishmania-induced cell-mediated immunosuppression as a predisposing factor for generalized demodicosis.