No description available.

Macrophage-derived inflammation contributes to chronic inflammation in adipose tissue in obesity and is also linked to the development of skeletal muscle (SM) insulin resistance. The long-chain n-3 PUFA have been shown to modulate cytokine secretion from macrophages, though subsequent effects on SM inflammation and function are unknown. A model of macrophage conditioned media (MCM) was used to examine effects of n-3 PUFA on macrophage inflammation and consequent effects on SM cells. Treatment of RAW 264.7 macrophages with long-chain n-3 PUFA decreased LPS-induced MCP-1 and IL-6 gene expression and MCP-1 secreted protein. In turn, MCM from n-3 PUFA-treated macrophages decreased TNF-α and IL-6 gene expression in LPS-stimulated L6 SM cells, but did not affect insulin-stimulated pAkt content. Long-chain n-3 PUFA did not affect gene expression of inflammatory signaling intermediates NF-κB and TLR4. Overall this thesis suggests that long-chain n-3 PUFA are important nutritional strategies for reducing macrophage-derived inflammation, with ensuing benefits in SM inflammation. / NSERC-CGS, Ontario Graduate Scholarship

Reproductive success depends on a functional oviduct for gamete storage, maturation, fertilization, and early-conceptus development. The ovarian-derived sex steroids estradiol and progesterone are known to affect functionality of the oviduct. Advances in microarray and NanoString technology allow for gene expression analysis to increase understanding of processes critical for fertility. Studies were conducted to investigate mechanisms regulating oviductal function in cattle and mice by using the Bovine Gene 1.0 ST array and the Mouse Gene 430-2.0 arrays (Affymetrix Inc., CA), respectively. For the first study, oviducts were collected from heifers assigned to luteal or follicular phase groups. In the second study oviducts were collected from immature mice with a global deletion of estrogen receptor-1 (ESR1) and their wild-type littermates at 23 days of age or 48 hr after treatment with 5 IU of PMSG. Following microarray hybridization, the resulting datasets were analyzed using Partek Genomics Suite 6.6 (Partek Inc., MO). The results of the first two studies illustrated a dynamic hormonal regulation of the oviductal epithelium and revealed the identity of novel genes affecting fertility in cattle and gave us insights into the genes regulated by estrogen and ESR1 in mice. Many genes identified as differentially regulated are believed to play an integral role in the regulation of oviductal inflammation. Therefore, the objective of the third study was to test the hypothesis that intraperitoneal administration of E. Coli-derived lipopolysaccharide induces the expression of inflammatory mRNAs in the mouse oviduct. Mice were treated with 0, 2 μg or 10 μg of LPS from E. Coli. and killed 24 h later. Oviducts were collected for determination of inflammatory gene expression by a targeted NanoString approach using the nCounter GX Mouse Inflammation Kit (NanoString Technologies, Wa). Results indicate that systemic treatment with LPS induces inflammation in the oviducts of mice and provides evidence of a repeatable animal model of oviductal inflammation. Overall, data from these studies extends our knowledge of the mechanisms regulating oviductal functions and immune response, as well as identified target molecules and processes to improve production animal and human fertility.