Polarization of chemoattractant receptor signaling during neutrophil chemotaxis

Morphologic polarity is necessary for chemotaxis of mammalian cells. As a probe of intracellular signals responsible for this asymmetry, the pleckstrin homology domain of the AKT protein kinase (or protein kinase B), tagged with the green fluorescent protein (PHAKT-GFP), was expressed in neutrophils. Upon exposure of cells to chemoattractant, PHAKT-GFP is recruited selectively to membrane at the cell's leading edge, indicating an internal signaling gradient that is much steeper than that of the chemoattractant. Translocation of PHAKT-GFP is inhibited by toxin-B from Clostridium difficile, indicating that it requires activity of one or more Rho guanosine triphosphatases.     

The immune response to dietary antigens and its influence on disease susceptibility in farm animals

Transient hypersensitivity reactions of the intestinal immune system to dietary antigens result in increases in enterocyte turnover and villous atrophy. These changes occur in the intestine of the post weaned piglet and precede the proliferation of E. coli and the development of post-weaning diarrhea. We therefore postulated that a transient cell mediated immune response to dietary antigens may increase susceptibility to disease. The interaction of dietary and microbial antigens upon the gut immune system has been investigated in mice and pigs and it has been shown that both exert powerful regulatory effects upon each other.     

Effect of aerosol challenge on a population of free lung cells in parenterally immunised calves

There is concern that parenteral respiratory vaccines may potentiate lung disease rather than protect against it. In vivo indicators of inflammation in calves were assessed to determine if vaccine-mediated inflammation occurs in the lung following aerosol challenge of parenterally immunised subjects. Preliminary results using a simple protein antigen suggest that changes in the free cell populations of the lung are likely to be a more sensitive indicator of immune-mediated inflammation than clinical parameters or peripheral changes in serum complement.     

Antibody production by the pig colon during infection with Treponema hyodysenteriae

When 47 pigs were dosed orally with cultures of Treponema hyodysenteriae, 44 (94 per cent) developed swine dysentery. Of those which recovered and were rechallenged, nine of 21 (43 per cent) showed clinical signs, as did one of 10 (10 per cent) challenged on a third occasion. Clinical disease was associated with development of specific IgG, IgA and IgM antibodies in serum and the local production of IgA in gut mucosal tissues. The appearance of antibody was not directly related to protection but rather indicated either prolonged exposure (in the case of serum IgG) or recent exposure to T hyodysenteriae (for secretory IgA). Infection also resulted in the appearance of IgG and IgA memory cells in gut-associated lymphoid tissue. However, these studies indicated that humoral immunity alone is not responsible for the onset of a protective response to T hyodysenteriae in the colon.     

Two genetic defects in alphaIIb are associated with type I Glanzmann's thrombasthenia in a Great Pyrenees dog: a 14-base insertion in exon 13 and a splicing defect of intron 13

Glannzmann's thrombasthenia (GT) is an autosomal recessive bleeding disorder caused by qualitative or quantitative deficiencies of the platelet membrane glycoprotein alphaIIbbeta3. This is the first report of a molecular genetic basis for type I GT in dogs. As previously reported, a thrombasthenic Great Pyrenees dog (dog No. 1) experienced uncontrolled epistaxis despite results of coagulation screening tests, platelet quantitation, and von Willebrand factor quantitation that were within reference ranges. Platelet aggregation was minimal in response to agonists. Flow cytometry, autoradiography, and immunoblot experiments demonstrated either marked reduction or absence of glycoproteins alphaIIb and beta3. In this study, we report the presence of a 14-base insertion in exon 13 and defective splicing of intron 13 in the alphaIIb gene of two thrombasthenic dogs (Nos. 1 and 8). The insertion disrupted the fourth alphaIIb calcium-binding domain, caused a shift in the reading frame and resulted in a premature termination codon. Possible consequences of this mutation include decreased alphaIIb mRNA stability and production of truncated alphaIIb protein that lacks the transmembrane and cytoplasmic domains and a large portion of the extracellular domain. We identified the dam, sire, and three littermates of dog No. 8 as carriers of the alphaIIb mutation. Canine alphaIIb and beta3 genes share significant homology with the genes in human beings, making canine GT an excellent translational model for human GT. A defined molecular basis for canine GT will enhance ongoing gene therapy research and increase the understanding of structure-function relationships of this integrin.     

A study of the discrete and interactive effects of different polysaccharides on the digestibility of diets fed to barramundi (Lates calcarifer)

This study examined the single, paired and combined inclusion effect of a range of different polysaccharide types on the dry matter, protein and energy digestibility of diets fed to barramundi (Lates calcarifer). The different polysaccharides included pregelled starch, cellulose, lignin and pectin. There were significant differences among the digestibility parameters of the diets with the different inclusion levels of each of the different polysaccharide types. Using a manova analysis, effects were noted for polysaccharide type, inclusion level and interaction terms on the digestibilities of dry matter, protein and energy. Cellulose addition resulted in a reduction in both dry matter and energy that was largely commensurate with its inclusion level, but its effect on protein digestibility was marginal. Starch had the least effect on any of the digestibility parameters of all the polysaccharide types examined. Pectin had the largest effect on dry matter, while lignin had the greatest impact on diet protein and energy digestion. In the diets with paired combinations of polysaccharides, lignin and pectin were responsible for negatively synergistic interactions in all digestibility parameters. These results show that different polysaccharide classes can have distinctly different effects on diet digestibility parameters.