The impact of habitually consuming a high protein (HP) diet at the upper limit of the acceptable macronutrient distribution range (AMDR) on kidney health is unknown. The current study was designed to test the hypothesis that long-term consumption of a diet providing 35% of energy as protein will have negative consequences on renal health, as assessed in a pig model. Methods: Adult female, non-pregnant, commercial pigs (Genesus) were randomized to receive either NP (15% energy from protein) or HP (35% energy from protein) isocaloric diets for either 4 or 8 months. Diets contained whole protein sources with an animal: plant ratio of 2:1 in the NP diet to mimic the average Canadian diet. The increased protein in the HP diet was achieved by increasing egg and dairy protein sources. Body composition was measured by dual-energy X-ray absorptiometry. Glomerular volume and kidney fibrosis were evaluated on kidney sections by quantitative image analysis. The inflammatory marker monocyte chemoattractant protein-1 (MCP-1) and the growth factor transforming growth factor beta-1(TGFβ1) were assessed in renal tissue using commercial ELISA kits. Results: Pigs given the HP diet had lower body weights and percentage of body fat. Pigs consuming the HP diet had significantly higher glomerular filtration rates (GFR) and larger kidneys. Renal MCP-1 levels and renal fibrosis also were significantly higher in pigs given the HP diet, while proteinuria and renal TGFβ1 expression did not differ. Conclusion: These findings suggest that, despite the potential benefit of the HP diet on body composition, long-term intakes of protein at the upper limit of the AMDR may compromise renal health in healthy female pigs. / October 2008

The impact of habitually consuming a high protein (HP) diet at the upper limit of the acceptable macronutrient distribution range (AMDR) on kidney health is unknown. The current study was designed to test the hypothesis that long-term consumption of a diet providing 35% of energy as protein will have negative consequences on renal health, as assessed in a pig model. Methods: Adult female, non-pregnant, commercial pigs (Genesus) were randomized to receive either NP (15% energy from protein) or HP (35% energy from protein) isocaloric diets for either 4 or 8 months. Diets contained whole protein sources with an animal: plant ratio of 2:1 in the NP diet to mimic the average Canadian diet. The increased protein in the HP diet was achieved by increasing egg and dairy protein sources. Body composition was measured by dual-energy X-ray absorptiometry. Glomerular volume and kidney fibrosis were evaluated on kidney sections by quantitative image analysis. The inflammatory marker monocyte chemoattractant protein-1 (MCP-1) and the growth factor transforming growth factor beta-1(TGFβ1) were assessed in renal tissue using commercial ELISA kits. Results: Pigs given the HP diet had lower body weights and percentage of body fat. Pigs consuming the HP diet had significantly higher glomerular filtration rates (GFR) and larger kidneys. Renal MCP-1 levels and renal fibrosis also were significantly higher in pigs given the HP diet, while proteinuria and renal TGFβ1 expression did not differ. Conclusion: These findings suggest that, despite the potential benefit of the HP diet on body composition, long-term intakes of protein at the upper limit of the AMDR may compromise renal health in healthy female pigs.

The impact of habitually consuming a high protein (HP) diet at the upper limit of the acceptable macronutrient distribution range (AMDR) on kidney health is unknown. The current study was designed to test the hypothesis that long-term consumption of a diet providing 35% of energy as protein will have negative consequences on renal health, as assessed in a pig model. Methods: Adult female, non-pregnant, commercial pigs (Genesus) were randomized to receive either NP (15% energy from protein) or HP (35% energy from protein) isocaloric diets for either 4 or 8 months. Diets contained whole protein sources with an animal: plant ratio of 2:1 in the NP diet to mimic the average Canadian diet. The increased protein in the HP diet was achieved by increasing egg and dairy protein sources. Body composition was measured by dual-energy X-ray absorptiometry. Glomerular volume and kidney fibrosis were evaluated on kidney sections by quantitative image analysis. The inflammatory marker monocyte chemoattractant protein-1 (MCP-1) and the growth factor transforming growth factor beta-1(TGFβ1) were assessed in renal tissue using commercial ELISA kits. Results: Pigs given the HP diet had lower body weights and percentage of body fat. Pigs consuming the HP diet had significantly higher glomerular filtration rates (GFR) and larger kidneys. Renal MCP-1 levels and renal fibrosis also were significantly higher in pigs given the HP diet, while proteinuria and renal TGFβ1 expression did not differ. Conclusion: These findings suggest that, despite the potential benefit of the HP diet on body composition, long-term intakes of protein at the upper limit of the AMDR may compromise renal health in healthy female pigs.

No description available.

With the absence of gut microbiota, gnotobiotic (Gn) pigs are a unique animal model for studying infection and immunity, and evaluating vaccine and therapeutics for human enteric pathogens. Here, we demonstrate Gn pigs as effective large animal models for human enteric viruses, through evaluating human enterovirus 71 (EV71) infection and immunity, and vaccine and therapeutics for human rotavirus (HRV). Gn pigs could be infected via oral or oronasal route, the natural route of infection. Infected pigs developed clinical signs including fever, neurological and respiratory signs, similar to those seen in human patients. Fecal shedding up to 18 days post infection and virus distribution in intestinal, respiratory and central nervous system tissues were observed. Strong mucosal and systemic T cell responses (IFN-γ producing CD4+ and CD8+ T cells) and systemic B cell responses (serum neutralizing antibodies) were also detected. The study demonstrates a novel large animal model for EV71 to investigate viral pathogenesis, immunity, and to evaluate vaccine and antiviral drugs. Using the well-established Gn pig model for HRV, the adjuvant and therapeutic effects of prebiotics rice bran (RB) and probiotics were evaluated. RB alone or RB plus probiotic Lactobacillus rhamnosus GG (LGG) and probiotic E. coli Nissle 1917 (EcN), were shown to protect against rotavirus diarrhea (80%-100% reduction in the incidence rate) significantly and display strong immune - stimulatory effects on the immunogenicity of an oral attenuated HRV (AttHRV) vaccine. Mechanisms for the adjuvant effect include stimulating the production of intestinal and systemic IFN-γ] producing T cells and promoting mucosal IgA antibody responses. The mechanisms for reducing rotavirus diarrhea include promoting LGG and EcN growth and colonization and host gut health, and maintaining gut integrity and permeability during rotavirus infection. We showed that RB plus LGG and EcN is a highly effective therapeutic regimen against HRV diarrhea. Together, these results indicated that Gn pigs may serve as an excellent animal model for the study of infection, immunity, vaccine and therapeutics for human enteric viruses. / Ph. D.

No description available.

No description available.

Cystic Fibrosis (CF) is a lethal autosomal recessive genetic disorder caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR transports anions at the apical surface of epithelial membranes and functions in many areas of the body. However in CF, loss of CFTR function in the lungs is the major source of morbidity and mortality. Replacing the defective CFTR in the lungs through gene therapy has the potential to cure the disease. Recombinant adeno-associated virus (AAV) is an effective gene transfer vector and has been used extensively to deliver genes to cells in culture. A number of clinical trials using AAV have been attempted for a variety of diseases, including CF, albeit with limited success. Poor vector transduction efficiency prevents effective gene therapy. We have previously used a technique to greatly increase the transduction efficiency of AAV in human lung tissues by selecting from a library of AAVs using a directed evolution technique. However, this evolution was performed in cultured cells and did not fully represent the in vivo environment in which the AAV would be used. In 2008, a CF pig model was developed to develop a further understanding of the mechanisms of CF and CFTR function. We hypothesized that we could use directed evolution to select for a vector in vivo using the pig, allowing gene therapy studies to be conducted in a physiologically relevant model of CF. We selected a novel AAV variant, called AAV2H22, which is closely related to AAV2 but with greatly increased transduction efficiency in pig airway epithelia. AAV2H22 displayed specific tropism for pig airway epithelia and saturated cell surface receptors, indicating specific binding in those cells. We found that AAV2H22-mediated gene transfer corrected chloride and bicarbonate transport defects both in vitro and in vivo. Importantly, bicarbonate transport was sufficient to normalize pH in the airway surface liquid, resulting in increased bacterial killing likely due to increased activity of antimicrobial peptides. To investigate the mechanics of the increased transduction of AAV2H22, capsid mutants were assayed for transduction efficiency. Two of the five amino acid differences between AAV2 and AAV2H22 lie at the surface and are predicted to alter capsid binding. This is consistent with the results showing specific binding in cultured airway epithelia. This research has important implications for gene therapy and investigations using AAV2H22 will increase our understanding of the biology needed to successfully treat CF.

The failure of β cells to secrete sufficient amounts of insulin is a key feature of diabetes mellitus. Each β cell secretes only a small amount of insulin upon stimulation in a highly regulated fashion: young insulin is preferentially released, whereas old insulin is mainly degraded within the β cell. How this process is regulated in vivo and likely altered in diabetes is currently unknown. We present here a transgenic pig model that allows the in vivo fluorescent labeling of age-distinct insulin secretory granule pools, hence providing a close-to-life readout of insulin turnover. This will enable the study of alterations in β cell function in an animal model close to humans.

No description available.