Project description:Caesarean-delivered preterm pigs were fed 3 d of parenteral nutrition followed by 2 d of enteral formula feeding. Antibiotics (n=11) or control saline (n=13) were given twice daily from birth to tissue collection at d 5. NEC-lesions and intestinal structure, function, microbiology and immunity markers were recorded. We used Affymetrix microarrays to investigate gene expression in intestinal tissues of preterm piglets treated with antibiotics or control saline. Twenty-four preterm piglets were delivered by caesarean section on day 105 of gestation from two healthy sows. All piglets were initially provided with parenteral nutrition via a vascular catheter, combined with small amounts of minimal enteral nutrition. On day three, all parenteral nutrition was stopped and total enteral nutrition was given through an oro-gastric feeding tube. Piglets were allocated into controls ( n=13) and an intervention group receiving oral and systemic broad-spectrum antibiotics ( n=11). To assure high systemic and intra luminal MIC values antibiotics were given both orally and intramuscularly. All antibiotics were given directly after feeding with an oral bolus and control pigs were given corresponding amounts of saline. On day five, all piglets were euthanized, and small intestinal tissue collected.
Project description:Oral food intake maintains gastrointestinal cell turnover and impacts the morphology and function of intestinal epithelial cells. However, the underlying mechanism is not fully elucidated, especially in the large intestine. Therefore, we analyzed the colonic epithelial cell turnover in starved and re-fed mice.
Project description:Oral food intake maintains gastrointestinal cell turnover and impacts the morphology and function of intestinal epithelial cells. However, the underlying mechanism is not fully elucidated, especially in the large intestine. Therefore, we analyzed the colonic epithelial cell turnover in starved and re-fed mice. EpCAM+CD45- colonic epithelial cells (ECs) in re-fed mice treated with or without antibiotics were isolated and sorted by FACS for RNA extraction and hybridization on Affymetrix microarrays.
Project description:The specific role of nutritional status during gestation in exacerbating or mitigating fetal toxicity of ethanol (EtOH) remains unclear. We used an intragastric infusion model (Total enteral nutrition, TEN) to appropriately control for dietary EtOH consumption and caloric intake. Time impregnated female Sprague Dawley rats (250-300 g) were surgically cannulated with intragastric cannulae (on gestation day 4) and infused either control or EtOH-containing diets. Rats were fed 220 kcal/ kg3/4/d (NRC recommended caloric intake for gestation) or 160 kcal/ kg3/4/d (undernourished to 72% of control) diets containing either 12 g/kg/d EtOH or an isocaloric amount of carbohydrates from gestation days 6-15. Maternal hepatic gene expression profiles were assessed on GD15. The present data dissect specific effects of nutritional status during gestation and EtOH intake and the interaction thereof. The data reveal a highly significant interaction between undernutrition and EtOH consumption. Keywords: Nutrition-gene interactions
Project description:Most commonly used models of non-alcoholic steatohepatitis (NASH) are diets based on specific gene knockouts or represent extreme manipulations of diet. We have examined the effects of modest increased caloric intake and high dietary unsaturated fat content on the development of NASH in male rats using a model in which overfeeding is accomplished via intragastric infusion of liquid diets as a part of total enteral nutrition. Male Sprague dawley rats were fed diets 5% corn oil containing diets at 187 Kcal/kg3/4/d or fed 70% corn oil containing diets at 220 Kcal/kg3/4/d for a period of 3 weeks. Hepatic gene expression were assessed at the end of the study. Our results indicate that overfeeding of high unsaturated fat diets leads to pathological, endocrine and metabolic changes characteristic of NASH patients and is associated with increased oxidative stress and TNF-a. Experiment Overall Design: Two groups of male sprague dawley rats were fed liquid diets via total enteral nutrition. Experiment Overall Design: Group 1, Control, Rats were fed diets containing 5% Corn oil at 187 Kcal/kg3/4/d for 3 weeks. Experiment Overall Design: Group 2, NASH, Rats were fed diets containing 70% corn oil at 220 Kcal/kg3/4/d for 3 weeks.
Project description:Purpose: Surgical treatment of congenital and neonatal intestinal diseases often requires resection of intestine and/or diverstion of luminal flow. Subsequent growth on enteral nutrition alone depends on adaptation of the remaining bowel. The role of mechanoluminal stimulation in driving intestinal adaptation and the effects of depriving the intestine of mechanoluminal stimulation are unknown. Methods: 5 pairs of intestine from neonatal surgical patients were obtained at ileostomy reversal. The proximal segment contiguous with enteral feeding was denoted “fed” and the distal segment without luminal flow was denoted “unfed". Following RNA extraction, these samples underwent deep sequencing. Results: Fed intestine had increased expression of genes involved in inflammation and immune regulation and steroid secretion. Unfed intestine had increased expression of genes involved in digestion and transport. Conclusion: Independent of external influences, the presence or absence of enteral mechanoluminal stimulation causes significant alterations in gene expression in intestine.
Project description:Since leptin signaling in the hypothalamus is critical to regulate food intake and body weight, we investigated how celastrol alters the hypothalamic transcriptome of DIO mice. By doing this analysis, genes with potential relevance for celastrol-mediated leptin sensitization could be identified.
Project description:Human monocytes are phagocytic leucocytes which circulate in the peripheral blood and play important roles in immunity and inflammation. They also represent circulating M-^QmetabolicM-^R sentinels in plasma, uptake lipids following food intake, and contribute to atherosclerotic plaque formation. We therefore investigated their homeostatic responses to food intake in vivo and to determine the response of human monocytes to food intake. We isolated monocyte subsets from the blood of healthy volunteers one hour before and four hours after a western type lunch.
Project description:The specific genes influencing the quantitative variation in macronutrient preference and food intake are virtually unknown. We refined a previously identified mouse chromosome 17 (MMU17) region harboring quantitative trait loci (QTL) with large effects on preferential macronutrient intake-carbohydrate (Mnic1), total kilcalories (Kcal2), and total food volume (Tfv1) using interval-specific congenic strains. These loci were isolated in the [C57BL/6J.CAST/EiJ-17.1-(D17Mit19-D17Mit50); B6.CAST-17.1] strain, developed by introgressing a ~40.1 Mb CAST MMU17 region into recipient B6 genome. In a diet selection paradigm (carbohydrate/protein vs. fat/protein), these B6.CAST-17.1 sub-congenic mice eat 30% more calories from the carbohydrate-rich diet, ~10% more total calories, and ~9% more total food volume per body weight. In the current study, this carbohydrate-preferring B6.CAST-17.1 subcongenic strain was crossed with the fat-preferring inbred B6 strain to generate a subcongenic-derived F2 mapping population; genotypes were determined using a high-density, custom SNP panel. The main outcome of this study is that genetic linkage analysis greatly reduced the 95% confidence interval (CI) for Mnic1 (encompassing Kcal2 and Tfv1) from 40.1 to 29.5 Mb and more precisely established the QTL boundaries. Specifically, the genetic architecture for Mnic1 (preferential carbohydrate intake) does not follow the same pattern as that for co-localized Kcal2/Tfv1 (total kcal and food volume, respectively), suggesting the presence of separate quantitative trait genes for these food intake traits. No genetic linkage for self-selected fat intake was detected, underscoring the carbohydrate-specific effects of this MMU17 locus. The Mnic1/Kcal2/Tfv1 QTL was further de-limited to a ~19.1 Mb interval, based on the absence of macronutrient diet selection phenotypes in subcongenic HQ17IIa mice that possess CAST MMU17 donor segment on a C57BL/6Jhg/hg background. A second key finding is the separation of two energy balance QTLs: Mnic1/Kcal2/Tfv1 for food intake and a newly discovered locus regulating short term body weight gain. The genes Decr2, Ppard and Agapt1 in the critical QTL interval were identified and prioritized using a combination of genome sequence analysis, and tag-based transcriptome sequencing to measure hypothalamic gene expression in non-recombinant F2 controls, possessing cast/cast and b6/b6 genotypes across the sub-congenic segment. Global gene expression profiles in the hypothalamus were compared between non-recombinant subcongenic-derived F2 mice, possessing a genotype of cast/cast (n=12) or b6/b6 (n=12) across the Chr 17 subcongenic segment and b6/b6 across the rest of the genome. RNA was isolated from individual mice of each genotype that were selected for study. Specifically, twelve mice of each genotype that displayed the most divergent phenotypic values for self-selected carbohydrate and total kcal intake were chosen for gene expression analysis, i.e., cast/cast mice with the highest and b6/b6 mice with the lowest kcal intake from carbohydrate, or the 25% tails of the distribution. Tissues were harvested ~48 h after re-initiation of the carbohydrate/protein vs. fat/protein diets, following an extended wash-out period on chow diet after completion of the 10 d macronutrient selection test.