Salmonella enterica serovar Typhi, a major public health concern in developing countries, continues to be a priority for the World Health Organization. S. Typhi possesses a viaB locus responsible for the biosynthesis of the Vi-capsular antigen, a significant virulence factor at the focus of developing improved prophylaxis for typhoid fever. Tissue culture experiments have demonstrated that S. Typhi wild-type capsule-expressing strain elicits less chemokine secretion than a viaB mutant. Calf experiments using the viaB mutant resulted in an increase inflammatory response. Osmolarity is one of the control signals that affect the biosynthesis of the Vi antigen. Under high osmolarity growth conditions of 300 mM and greater, Vi production is suppressed and S. Typhi is highly invasive. Studies reveal that the viaB mutant displays increased invasion towards intestinal epithelial cells. Our first objective was to implement direct and indirect methods to localize and detect Vi expression within intestinal epithelial cells and bovine Peyer's patch. The second objective was to compare the invasiveness between a viaB mutant, an ompR mutant, and S. Typhi grown under hyperosmolarity. We also measured the effects of these strains in eliciting inflammation in the calf model. We report that tviB was significantly up regulated intracellularly within T84 polarized cells. In the calf experiments, tviB was expressed at levels significantly higher in calf tissue following invasion compared to inoculum grown under Vi-suppressing conditions. Together, these results support the idea that the Vi capsular antigen is expressed after invasion of intestinal epithelial cells in vivo. We found that S. Typhi grown under high osmolarity, the viaB mutant, and the ompR mutant had increased invasion in polarized T84 cells and bovine ileal tissue. Fluid accumulation among Vi-deficient and Vi-suppressed strains was similar. The histopathology of the inflammatory lesions of the small intestine produced by the Vi-deficient and suppressed strains was quite comparable. Our data supports the notion that Vi-suppressed and Vi mutants of S. Typhi exhibit similar levels of increased invasion and inflammation, perhaps mechanistically through the inactivation of the Vi antigen.

Le desserrage des tiges est une complication fréquente des plâtres avec tiges transcorticales (TP) chez les grands animaux, nécessitant souvent leur retrait prématuré avant la guérison des fractures. Les charges excessives centrées sur le cortex à l’interface os-tige proximo-externe et disto-interne causent de l'ostéolyse. En utilisant un modèle de veau nouveau-né, ce projet a évalué un nouveau système de tige-manchon et anneau integré dans un plâtre (PS) optimisé pour réduire la contrainte péri-implant et le stress à l'interface os-implant. On a émis l'hypothèse que les PS se traduiraient par une ostéolyse péri-implant moindre par rapport aux TP. Dix veaux en bonne santé, de 3 semaines d'âge, ont été implantés avec les TP ou PS dans le métacarpe droit, à raison de 2 implants par veau. Les veaux ont été observés quotidiennement pour le confort et la boiterie et ont été euthanasiés à 28 jours. Les données recueillies comprenaient les radiographies à la chirurgie et à l'euthanasie et les mesures histomorphométriques de contact os-implant sur des échantillons non-décalcifiés avec les implants in situ. Les données ont été analysées en utilisant le test de Cochran-Mantel-Haenszel, une valeur de P <0,05 a été considérée comme significative. L'épaisseur corticale était plus importante pour les implants distaux que proximaux pour les deux groupes lors de la chirurgie (P = 0,03), mais était similaire entre les groupes (P > 0,3). Les veaux avec TP ont développé une boiterie plus tôt (au jour 21) que les veaux avec PS (P = 0,04). Histologiquement, il y avait plus de contact direct os-implant cortical pour les implants PS distaux que les implants TP (P = 0,04). La jonction métaphyso-diaphysaire osseuse où les implants proximaux étaient situés est impropre aux deux systèmes; chacun a un minimum de contact os-implant et de l'ostéolyse extensive. Le système PS n'ayant pas causé une ostéolyse importante lorsque implantés dans l'os diaphysaire et peut-être une alternative convenable aux TP pour des fractures comminutives des membres distaux. / Pin loosening is a common complication of transfixation pincasts (TP) in large animals, often necessitating premature removal before fracture healing. The excessive loads centered on the proximo-external and disto-internal cortices of the bone-pin interface cause osteolysis. Using a neonatal calf model, this project evaluated a novel pin-sleeve and ring cast system (PS) optimized to decrease peri-implant strain and evenly share stress at the bone-implant interface. It was hypothesized that PS would result in less peri-implant osteolysis compared to TP. Ten, 3-week-old, healthy calves were implanted with either TP or PS in the right metacarpus, 2 implants per calf. Calves were scored daily for lameness and were euthanized at day 28. Collected data included radiographs at surgery and euthanasia and histomorphometric measures of bone-implant contact on non-decalcified specimens with the implants in situ. Data was analyzed using Cochran-Mantel-Haenszel test; a P-value <.05 was considered significant. The cortical thickness was larger for distal implants than proximal implants for both groups at surgery (P = 0.03), but were similar between groups (P > 0.3). TP calves developed lameness sooner, at day 21, than PS calves (P = 0.04). Histologically, there was more direct cortical bone-implant contact for PS distal implants than TP implants (P = 0.04). The metaphyseal-diaphyseal junction where the proximal implants were situated is unsuitable bone for either system; each had minimal bone-implant contact and extensive osteolysis. The PS system did not cause significant osteolysis when instrumented in diaphyseal bone and is a suitable alternative to TP for comminuted distal limb fractures.

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