Dietary lipids alter the effect of steroids on the transport of fructose following intestinal resection in rats.

BACKGROUND: Glucocorticosteroids alter intestinal morphology and transport. We tested the hypothesis that the desired intestinal adaptive response following intestinal resection may be enhanced further by the locally active steroid budesonide, and by feeding a saturated as compared with a polyunsaturated fatty acid diet. METHODS: An in-vitro uptake method was used to assess intestinal fructose uptake by rats of semisynthetic diets enriched in saturated or polyunsaturated fatty acids, and injected with budesonide or control solution. RESULTS: Budesonide increased ileal fructose uptake in chow and PUFA-fed animals, but reduced jejunal fructose uptake in rats fed SFA. GLUT5 and GLUT2 protein and mRNA did not correlate with changes in fructose uptake. Steroids reduced jejunal proglucagon expression in animals fed chow. Animals fed SFA and given budesonide had a reduction in jejunal ODC mRNA compared with those fed PUFA or chow. CONCLUSIONS: (1) budesonide increases ileal fructose uptake following intestinal resection, and this beneficial effect is prevented by feeding SFA rather than PUFA; (2) fructose uptake does not correlate with GLUT5 and GLUT2 protein and mRNA; (3) ODC and proglucagon may be involved in this adaptive response.

Effects of breakfast meal composition on second meal metabolic responses in adults with Type 2 diabetes mellitus.

OBJECTIVE: We tested the relative importance of a low-glycemic response versus a high glycemic response breakfast meal on postprandial serum glucose, insulin and free fatty acid (FFA) responses after consumption of a standardized mid-day meal in adult individuals with Type 2 diabetes mellitus (DM). DESIGN: Following an overnight fast of 8-10 h, a randomized crossover intervention using control and test meals was conducted over a 3-week-period. A fasting baseline measurement and postprandial measurements at various time intervals after the breakfast and mid-day meal were taken. SUBJECTS: Forty-five Type 2 DM subjects completed the requirements and were included in the study results. INTERVENTIONS: Two different breakfast meals were administered during the intervention: (A) a high glycemic load breakfast meal consisting of farina (kJ 1833; carbohydrate (CHO) 78 g and psylium soluble fiber 0 g), (B) a low-glycemic load breakfast meal consisting of a fiber-loop cereal (kJ 1515; CHO 62 g and psyllium soluble fiber 6.6 g). A standardized lunch was provided approximately 4 h after breakfast. Blood plasma concentrations and area under the curve (AUC) values for glucose, insulin and FFA were measured in response to the breakfast and mid-day lunch. Statistical analyses were performed using SAS software (8.02). Comparisons between diets were based on adjusted Bonferroni t-tests. RESULTS: In post-breakfast analyses, Breakfast B had significantly lower area under the curve (AUC) values for plasma glucose and insulin compared to Breakfast A (P<0.05) (95% confidence level). The AUC values for FFA were higher for Breakfast B than for Breakfast A (P<0.05) (95% confidence level). Post-lunch analyses indicated similar glucose responses for the two breakfast types. Insulin AUC values for Breakfasts B were significantly lower than Breakfast A (P<0.05) (95% confidence level). The AUC values for FFA were unaffected by breakfast type. CONCLUSIONS: These data indicate that ingesting a low-glycemic load meal containing psyllium soluble fiber at breakfast significantly improves the breakfast postprandial glycemic, insulinemic and FFA responses in adults with Type 2 DM. These data revealed no residual postprandial effect of the psyllium soluble fiber breakfast meal beyond the second meal consumed. Thus, there was no evidence of an improvement postprandially in the glycemic, insulinemic and FFA responses after the consumption of the lunch meal.

The effects of fiber enrichment of pasta and fat content on gastric emptying, GLP-1, glucose, and insulin responses to a meal.

OBJECTIVE: To assess whether the addition of viscous fiber at an amount recommended by the US FDA to allow a 'low saturated fat, cholesterol, soluble fiber and coronary heart disease', health claim label on a food package (1.7 g psyllium) and/or fat (30 g sunflower oil and 3 g sodium propionate) to a pasta meal would affect gastric emptying, postprandial glucose, insulin and GLP-1 concentrations. DESIGN: Ten subjects participated in a two-by-two single blind randomized crossover study. Four meals containing 50 g of available carbohydrate were consumed: pasta with or without psyllium enrichment served with a tomato sauce with (520 kcal per meal) and without (240 kcal per meal) fat. Blood samples were taken for 240 min following the meal and all subjects consumed a buffet meal at the end of the study. Gastric emptying was measured using the paracetamol absorption test. Blood was analysed for glucose, insulin, GLP-1. Visual analog scales were used to record feelings of hunger, pleasantness and nausea. RESULTS: The psyllium-enriched pasta had no significant effect on gastric emptying or the incremental area under the curve (IAUC) for GLP-1, insulin or glucose compared with the control pasta. The addition of polyunsaturated fat and sodium propionate significantly increased the IAUC for GLP-1 (P<0.001), delaying gastric emptying (P<0.002), and decreasing glucose (P<0.002). CONCLUSIONS: A dose of 1.7 g psyllium did not evoke measurable effects on gastric emptying, postprandial GLP-1, insulin or glucose metabolism. However the addition of 30 g of oil and 3 g of sodium propionate to the pasta did reduce gastric emptying, increase GLP-1 and reduce glucose and insulin concentrations. While this short-term study may have implications in terms of reducing the risk of diabetes and improving coronary risk factor profiles the long term effects of these nutrients need to be studied.

The locally acting glucocorticosteroid budesonide enhances intestinal sugar uptake following intestinal resection in rats.

BACKGROUND AND AIMS: Locally and systemically acting corticosteroids alter the morphology and transport function of the intestine. This study was undertaken to assess the effect of budesonide, prednisone, and dexamethasone on sugar uptake. METHODS: Adult male Sprague Dawley rats underwent transection or resection of 50% of the middle portion of the small intestine, and in vitro uptake of sugars was measured. RESULTS: The 50% enterectomy did not alter jejunal or ileal uptake of glucose or fructose. Prednisone had no effect on the uptake of glucose or fructose in resected animals. In contrast, in resected rats budesonide increased by over 120% the value of the jejunal maximal transport rate for the uptake of glucose, and increased by over 150% ileal uptake of fructose. Protein abundance and mRNA expression of the sodium dependent glucose transporter in brush border membrane (SGLT1), sodium independent fructose transporter in the brush border membrane (GLUT5), sodium independent glucose and fructose transporter in the basolateral and brush border membranes (GLUT2), and Na(+)/K(+) ATPase alpha1 and beta1 did not explain the enhancing effect of budesonide on glucose or fructose uptake. Budesonide, prednisone, and dexamethasone reduced jejunal expression of the early response gene c-jun. In resected animals, expression of the mRNA of ornithine decarboxylase (ODC) in the jejunum was reduced, and corticosteroids reduced jejunal expression of the mRNA of proglucagon. CONCLUSIONS: These data suggest that the influence of corticosteroids on sugar uptake in resected animals may be achieved by post translational processes involving signalling with c-jun, ODC, and proglucagon, or other as yet unknown signals. It remains to be determined whether budesonide may be useful to stimulate the absorption of sugars following intestinal resection in humans.

Intestinal resection- and steroid-associated alterations in gene expression were not accompanied by changes in lipid uptake.

BACKGROUND/AIMS: Glucocorticosteroids alter the morphology and transport function of the intestine of adult rats. This study was undertaken to assess the possible effect on intestinal lipid uptake of the locally acting steroid budesonide, or the systemically active prednisone or dexamethasone. METHODS: Sprague-Dawley rats underwent intestinal transection or 50% intestinal resection. Budesonide, prednisone, dexamethasone, or control vehicle was given for 2 weeks from the time of surgery. Uptake was measured using ring uptake technique. RESULTS: Resection had no effect on the mRNA expression for the early response genes, for proglucagon, or for the ileal lipid binding protein (ILBP), but was associated with reduced jejunal ornithine decarboxylase (ODC) mRNA and with reduced jejunal mRNA for the liver fatty acid binding protein (L-FABP). All three steroids reduced jejunal mRNA for proglucagon and c-jun, and did not affect the mRNA for L-FABP or for ILBP. These resection- and steroid-associated changes in gene expression were not associated with alterations in the intestinal uptake of long chain fatty acids or cholesterol. CONCLUSIONS: The resection-associated alterations in the RNA expression of ODC and L-FABP and the steroid-associated changes in mRNA expression of c-jun and proglucagon were not accompanied by variations in lipid uptake.

Dietary lipids alter the effect of steroids on the uptake of lipids following intestinal resection in rats.

Steroids alter the transport function of the intestine. This study was undertaken to assess the effect of glucocorticosteroids on lipid uptake in rats fed either a saturated (SFA) or a polyunsaturated fatty acid (PUFA) diet. Sprague-Dawley rats underwent transection or 50% resection of the small intestine. The steroids had no effect on the uptake of lipids. However, resection decreased the jejunal uptake of palmitic acid in animals fed SFA and increased the jejunal uptake of palmitic and linoleic acids in those fed PUFA. In animals undergoing intestinal resection, fed SFA, and given control vehicle, there was a reduction in jejunal proglucagon mRNA expression as compared to those fed chow or PUFA. Ornithine decarboxylase (ODC) mRNA expression in the jejunum of resected animals was reduced. In summary, dietary lipids modify the uptake of lipids in resected animals and ODC and proglucagon may be involved in this adaptive response.

Candidate foods in the Asia-Pacific region for cardiovascular protection: relevance of grains and grain-based foods to coronary heart disease.

This review elucidates the importance of healthy dietary and lifestyle habits to reduce morbidity and mortality associated with coronary heart disease (CHD), stroke and cardiovascular diseases. Given published evidence of the poor compliance, increased cost, and decreased benefit/risk ratios of medical therapies, individuals (and populations) are encouraged to adopt healthy life habits. The three most atherogenic dietary risk factors are saturated fat, cholesterol and obesity. Dietary patterns associated with the consumption of grains and grain-based foods predict risk of CHD independently of other life habits. Epidemiological and intervention studies elucidating the strong protective associations of grains, cereal fibers and anti-oxidant vitamins on CHD are reviewed. In summary, the consumption of grains and grain-based cereals is repeatedly associated with the ingestion of many nutrients, e.g., dietary fiber and anti-oxidants, that alter energy balance and nutrient intakes to positively affect cardiovascular health, especially when combined with healthy life habits,

Comparison of high- and low-glycemic-index breakfast cereals with monounsaturated fat in the long-term dietary management of type 2 diabetes.

BACKGROUND: Results of 6-wk studies suggest that high-carbohydrate diets are deleterious for people with type 2 diabetes. OBJECTIVE: Our objective was to see whether long-term replacement of dietary monounsaturated fatty acids (MUFAs) with carbohydrate from breakfast cereals with either a high or a low glycemic index (GI) affected blood glucose and lipids in subjects with type 2 diabetes. DESIGN: Subjects with type 2 diabetes (n = 91) were randomly assigned to receive approximately 10% of energy from a low-GI breakfast cereal, a high-GI cereal, or oil or margarine containing MUFA for 6 mo. Eating breakfast cereal was prohibited for subjects in the MUFA group. RESULTS: Seventy-two subjects completed the trial. The subjects who received cereals consumed approximately 10% more energy from carbohydrate than did the subjects in the MUFA group. Changes in glycated hemoglobin, body weight, and fasting cholesterol and triacylglycerol did not differ significantly among groups. HDL cholesterol increased by approximately 10% in the MUFA group compared with subjects who consumed either high- or low-GI cereals (P = 0.002). The ratio of total to HDL cholesterol was higher in the subjects who consumed the high-GI cereal than in the MUFA group at 3 mo but not at 6 mo (diet x time interaction, P = 0.041). During 8-h metabolic profiles, mean plasma insulin was higher and mean free fatty acids were lower in the 2 cereal groups than in the MUFA group (P < 0.05). CONCLUSIONS: A 10% increase in carbohydrate intake associated with breakfast cereal consumption had no deleterious effects on glycemic control or blood lipids over 6 mo in subjects with type 2 diabetes. The increase in plasma insulin and the reduction in free fatty acids associated with higher carbohydrate intake may reduce the rate of progression of diabetes.

Luminal amino acids acutely decrease intestinal mucosal protein synthesis and protease mRNA in piglets.

Because parenteral feeding is associated with negative N balance and reduced rates of protein synthesis in intestinal mucosa, we hypothesized that luminal exposure to specific amino acids or energy fuels would stimulate intestinal protein synthesis. We studied the acute effects of luminal nutrients on mucosal protein synthesis in the absence of systemic influences. Multiple jejunal segments constructed in piglets deprived of food overnight (n = 6) were randomly assigned to luminal perfusion with saline, 30 mmol/L amino acid mixture with or without 50 mmol/L glucose, or 30 mmol/L glutamine for 90 min. Protein synthesis was then measured by luminal perfusion with L-[2,6-(3)H]-phenylalanine. Energy substrates (glucose, short-chain fatty acids or beta-hydroxybutyrate) had no effect on mucosal protein synthesis. Relative to saline, a 30 mmol/L amino acid mixture or 30 mmol/L glutamine suppressed mucosal protein synthesis by 20-25% (P < 0.05). On the basis of these surprising results, we speculated that a coordinate reduction of proteolytic processes would be required to maintain positive intestinal N balance. Although intestinal protein catabolism cannot be assessed directly, the 30 mmol/L amino acid mixture acutely suppressed mucosal levels of mRNA encoding ubiquitin, 14-kDa ubiquitin conjugating enzyme and the C9 subunit of the proteasome by 20-30% (P < 0.05), demonstrating the sensitivity of components of the ATP-ubiquitin proteolytic pathway to acute regulation by nutrients. The suppression of protein synthesis by luminal amino acids in the absorptive state might lower intestinal utilization of amino acids to ensure efficient allocation of absorbed nutrients to nonintestinal tissues.

Jejunal mucosal protein synthesis: validation of luminal flooding dose method and effect of luminal osmolarity.

To validate a system to study acute regulation of protein synthesis in intestinal mucosa by luminal nutrients, we compared the fractional rate of protein synthesis (Ks) in jejunal mucosa using the intravenous flooding dose technique with the administration of a comparable concentration and specific activity of tracer in a luminal perfusate. Routes of tracer administration and surgery and perfusion trauma had no effect on mucosal Ks. Furthermore, four 10-cm jejunal segments (within a piglet) simultaneously but separately perfused with a luminal flooding dose had similar Ks values (mean, 43 +/- 2%/day; P > 0.05). Nutrient solutions perfused through four intestinal segments within an animal did not affect plasma levels of most amino acids or glucose. Because cellular hydration is important in regulating metabolism, the effects of physiological variation in luminal osmolarity were studied. Luminal osmolarities between 250 and 380 mosM did not affect mucosal Ks. The system described allows multiple comparisons within an animal and provides a robust model to study acute modulation of protein synthesis in intestinal mucosa by luminal stimuli.

The fermentable fiber content of the diet alters the function and composition of canine gut associated lymphoid tissue.

The ingestion of plant fibers and their susceptibility to microbial fermentation in the large bowel modulate intestinal morphology but little is known about effects on the gut associated lymphoid tissue (GALT). The aim of the present study was to determine the effect of consuming diets containing different levels of fermentability fiber on immune function. Sixteen adult mongrel dogs (23 +/- 2 kg) were fed (14 days) in a randomized cross over design two isoenergetic isonitrogenous diets containing 8.3 g/kg non-fermentable or 8.7 g/kg fermentable fibers. Lymphocytes were isolated from blood prior to starting the study and at the end of each diet period. At study completion, lymphocytes were isolated from the gut associated lymphoid tissue (GALT) of the small intestine for characterization by immunofluorescence and to determine their ability to respond to mitogenic stimulation. Feeding high fermentable fibers increased (P < 0.05) the CD4/CD8 ratio and decreased (P < 0.05) the proportion of B cells in peripheral blood without changing natural killer cell activity or the response to mitogens. Mesenteric lymph node cells from dogs fed the low then high fermentable fiber diet contained a higher (P < 0.05) proportion of CD4+ cells and a higher (P < 0.05) response to mitogens. Intraepithelial, Peyer's patches and lamina propria cells contained a greater (P < 0.05) proportion of CD8+ cells when dogs were fed a low fermentable fiber diet followed by a high fermentable fiber diet. T cell mitogen responses in vitro were higher for intraepithelial but lower for Peyer's patches and lamina propria cells from dogs who were fed the low fermentable fiber diet followed by the high fermentable fiber diet (P < 0.05). In conclusion, the fermentable fiber content of the diet had very little effect on the type and function of immune cells in peripheral blood. However, feeding dogs a high fermentable fiber diet for 2 weeks (after 2 weeks of consuming a low fermentable fiber diet) altered the T-cell composition of GALT and produced a higher mitogen response in the predominantly T cell tissues and a lower response in areas involved in B cell functions. In conclusion, the level of fermentable fiber in the diet appears to alter GALT properties. Further studies are required to determine the direct contribution of a high or low fiber diet to these changes and the physiological implications to the health of the animal.

Fermentable dietary fiber increases GLP-1 secretion and improves glucose homeostasis despite increased intestinal glucose transport capacity in healthy dogs.

Ileal proglucagon gene expression and postprandial plasma concentrations of proglucagon-derived peptides are reported to change with the type and quantity of dietary fiber ingested by rats. Within the intestine, proglucagon encodes several proglucagon-derived peptides known to modulate intestinal absorption capacity and pancreatic insulin secretion. To determine whether the chronic ingestion of fermentable dietary fiber regulates the expression and synthesis of proglucagon-derived peptides in the distal intestine to modulate glucose homeostasis, the following study was conducted: 16 adult dogs (23 +/- 2 kg) were fed isoenergetic, isonitrogenous diets containing a mixture of high fermentable dietary fibers (HFF) or low fermentable (LFF) wood cellulose for 14 d in a randomized cross-over design. Food was withheld for 16 h before an oral glucose tolerance test was conducted supplying 2 g of glucose/kg body wt, and peripheral blood was collected via a hind-leg catheter at 0, 15, 30, 45, 60, 90 and 120 min for plasma glucose, insulin and glucagon-like peptide-1(7-36)NH2 (GLP-1) analyses. Intestinal samples were collected after the second dietary treatment. Ileal proglucagon mRNA, intestinal (GLP-1) concentrations and the integrated area under the curves (AUC) for plasma GLP-1 and insulin were greater and plasma glucose AUC was reduced when dogs were fed the HFF diet compared to the LFF diet (P < 0.05). Intestinal villi heights, brush border and basolateral glucose transporter protein abundance and jejunal transport capacities were significantly greater when dogs were fed the HFF diet than when fed the LFF diet. In conclusion, improvements in glucose homeostasis are observed in healthy dogs when they ingest fermentable fibers.

Systemic short-chain fatty acids rapidly alter gastrointestinal structure, function, and expression of early response genes.

Luminal and systemic short chain fatty acids (SCFA) stimulate mucosal proliferation but the mechanism(s) is unclear. This study examined acute effects of systemic SCFAs on gastrointestinal structure and function and signals potentially mediating SCFA-induced mucosal proliferation. Male Sprague-Dawley rats (246+/-2 g) received nutrients as either standard total parenteral nutrition (TPN) or an isoenergetic, isonitrogenous formulation containing SCFAs (TPN + SCFA). Animals were randomized to one of five treatments: standard TPN for 72 hr, TPN + SCFA for 72 hr, or standard TPN followed by TPN + SCFA for the final 6, 12, and 24 hr. SCFAs reduced (P < 0.003) ileal protein within 6 hr. Jejunal GLUT2 expression was increased (P=0.0001) in all SCFA groups and ileal GLUT2 protein in the 6-, 12-, and 24-hr SCFA groups (P < 0.05). SCFAs increased (P < 0.003) ileal proglucagon abundance following 6, 12, and 24 hr, and plasma GLP-2 concentration following 12 hr (P < 0.03). Jejunal c-myc expression was increased (P < 0.001) following 6, 12, and 24 hr of SCFAs. SCFAs increased ileal c-myc, c-jun, and c-fos expression following 24 hr (P < 0.02), 12 hr (P < 0.05) and 6, 12, and 24 hr (P=0.0001), respectively. In conclusion, systemic SCFAs increase plasma GLP-2 and ileal proglucagon mRNA, GLUT2 expression and protein, and c-myc, c-jun, and c-fos expression.

Proglucagon and glucose transporter mRNA is altered by diet and disease susceptibility in 30-day-old biobreeding (BB) diabetes-prone and normal rats.

Diet is an important factor influencing the development of diabetes in the Biobreeding (BB) rat. Changes in gut development and absorption of nutrients in the diabetes-prone rat (Bbdp) and the subsequent effect on pancreatic function may play a role in the ultimate development of the disease. BBdp and normal (BBn) dams were fed one of three diets, chow or semipurified diets containing either soy or casein as the protein source. Pups were weaned at 21 d onto the same diet as their respective dam and killed at 30 d. Chow-fed BBn animals weighed significantly more than casein- or soy-fed BBn animals. Chow-fed BBdp had significantly greater small intestine and colon weight when expressed on a per body weight basis than BBn. With all three diets, BBdp animals had significantly lower colonic proglucagon mRNA abundance than did BBn animals. Adjusting for total colonic RNA content resulted in only casein- and soy-fed BBdp animals demonstrating significantly lower colonic proglucagon mRNA abundance than did BBn animals. BBn animals had higher levels of sodium-dependent D-glucose cotransporter (SGLT)-1 and sodium-independent glucose transporter (GLUT) 5 mRNA than did BBdp. Within BBn animals, casein and soy produced significantly lower levels of SGLT-1 and GLUT5 mRNA than did chow. This study demonstrates that BBn and BBdp animals respond differently to chow versus semipurified weaning diets. These differences may explain the differences in growth and intestinal development among the BBdp and BBn strains weaned onto different diets.

Short-chain fatty acid-supplemented total parenteral nutrition alters intestinal structure, glucose transporter 2 (GLUT2) mRNA and protein, and proglucagon mRNA abundance in normal rats.

Intestinal adaptation is a complex physiologic process that is not completely understood. Intravenous short-chain fatty acids (SCFAs) enhance intestinal adaptation after 80% enterectomy in rats. The purpose of this study was to examine rapid responses to SCFA-supplemented total parenteral nutrition (TPN) in the normal small intestine. After jugular catheterization, 31 Sprague-Dawley rats (weighing 258 +/- 3 g) were randomly assigned to receive standard TPN or an isoenergetic, isonitrogenous TPN solution supplemented with SCFAs (TPN+SCFA). Intestinal samples were obtained after 24 or 72 h of nutrient infusion. TPN+SCFA for 24 h increased (P < 0.05) the ileal RNA concentration (microg RNA/mg ileum) whereas TPN+SCFA for 72 h increased (P < 0.05) the ileal DNA concentration (microg DNA/mg ileum) and decreased (P < 0.05) the ileal protein concentration (microg protein/mg ileum). Ileal proglucagon mRNA abundance was elevated (P < 0.05) after 24 h of TPN+SCFA infusion and returned to levels seen with control TPN by 72 h. Glucose transporter 2 (GLUT2) mRNA was significantly higher (P < 0.05) in the TPN+SCFA groups at both time points when compared with control TPN groups. Ileal GLUT2 protein abundance in the 72-h TPN+SCFA group was significantly higher (P < 0.05) than that of all other groups. Sodium-glucose cotransporter (SGLT-1) mRNA and protein abundance and uptake of D-fructose and D-glucose did not differ between groups. Jejunal uptake of L-glucose and lauric acid was significantly higher (P < 0.05) after 72 h of TPN+SCFA than after 24 h, whereas the 24- and 72-h TPN groups did not differ. In summary, SCFAs led to rapid changes in ileal proglucagon and glucose transporter expression in rats receiving TPN and provide insights into therapeutic management of individuals with short bowel syndrome or intestinal malabsorption syndromes.

Kinetic analysis of L-glutamine transport into porcine jejunal enterocyte brush-border membrane vesicles.

L-Glutamine transport into porcine jejunal enterocyte brush border membrane vesicles was studied. Uptake was mediated by a Na(+)-dependent and a Na(+)-independent pathway as well as by diffusion. The initial rates of glutamine uptake over a range of concentrations is both Na(+)-gradient and Na(+)-free conditions were analyzed and kinetic parameters were obtained. Na(+)-dependent glutamine transport had a K(m) of 0.77 +/- 0.16 mM and a Jmax of 70.7 +/- 5.8 pmol mg protein-1 s-1; Na(+)-independent glutamine transport had a K(m) of 3.55 +/- 0.78 mM and a Jmax of 55.1 +/- 6.6 pmol mg protein-1 s-1. The non-saturable component measured with HgCl2-poisoned brush border membrane vesicles in the Na(+)-free condition contained passive diffusion and non-specific membrane binding and was defined to be apparent glutamine diffusion and the glutamine permeability coefficient (Kdiff) was estimated to be Kdiff = 3.78 +/- 0.06 pmol 1 mg protein-1 mmol-1 s-1. Results of inhibition experiments showed that Na(+)-dependent glutamine uptake occurred primarily through the brush border system-B degree transporters, whereas Na(+)-independent glutamine uptake occurred via the system-L transporters. Furthermore, the kinetics of L-leucine and L-cysteine inhibition of L-glutamine uptake demonstrated that neutral amino acids sharing the same brush border transporters can effectively inhibit each other in their transport.

Intestinal nutrient-gene interaction: the effect of feed deprivation and refeeding on cholecystokinin and proglucagon gene expression.

We tested the hypothesis that dietary components reaching the bovine small intestine influence the expression of genes that encode the gastrointestinal neuropeptides cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1). The amount of digesta reaching the intestine was manipulated during the experiment by withholding feed from five heifers fitted with ruminal, duodenal, and ileal cannulas for 48 h and then subsequent refeeding. Duodenal and ileal biopsies were collected using a fiber-optic endoscope. A Northern hybridization procedure was used to evaluate changes in gene expression. Blood concentrations of CCK and GLP-1 were determined with RIA. The data indicate that CCK blood concentration and mRNA abundance decreased during the period of feed deprivation, but they returned to predeprivation values within 16 to 24 h of refeeding. The GLP-1 blood concentration also decreased during feed deprivation and returned to predeprivation values within 4 to 8 h of refeeding, despite the fact that proglucagon mRNA abundance did not change significantly during feed deprivation and refeeding. These findings provide evidence that CCK and GLP-1 are released in response to nutrients that reach the small intestine and may be involved in the physiological process of digestion and possibly play a role in regulating feed intake in ruminants.

Glutamine supplementation maintains intramuscular glutamine concentrations and normalizes lymphocyte function in infected early weaned pigs.

Numerous studies in humans and rats have shown that glutamine supplementation during stressful conditions has favorable outcomes. However, the requirements for glutamine during weaning are unknown. Thus, the effects of glutamine supplementation in healthy and infected weaned pigs were investigated. At 21 d of age, pigs were weaned to an elemental diet supplemented with glutamine (+Gln) or an isonitrogenous diet containing nonessential amino acids (-Gln). At 26 d of age, pigs were intraperitoneally injected with Escherichia coli (+Ecoli) or buffered saline (-Ecoli) and killed at 28 d of age. Infection decreased (P < 0.05) plasma and intramuscular glutamine concentrations, but infected pigs that received +Gln diets had higher intramuscular glutamine levels than those that received -Gln diets. Infected pigs had elevated (P < 0.05) total leukocyte counts, and blood lymphocyte responses ([3H]-thymidine incorporation) to a mixture of phorbol myristate acetate and ionomycin were reduced. White blood cell counts were greater (P < 0.05) in +Gln than -Gln pigs. The peak responses to concanavalin A (Con A) by lymphocytes of +Ecoli+Gln pigs were greater (P < 0.05) than those of +Ecoli-Gln pigs and not different than those of noninfected pigs. Hence, glutamine supplementation maintained muscular glutamine concentrations and normalized lymphocyte function in infected pigs.

A physiological level of rhubarb fiber increases proglucagon gene expression and modulates intestinal glucose uptake in rats.

Previous work demonstrated that a high fiber diet upregulates proglucagon mRNA and secretion of glucagon-like peptide-1 [GLP-1(7-37)] and insulin compared with an elemental fiber-free diet. This study examined whether similar intakes of fibers differing in physiochemical and fermentative properties alter the expression of intestinal hormones and intestinal absorptive properties. Sprague-Dawley rats were fed either a 50 g/kg cellulose or rhubarb fiber diet for 14 d. Ileal proglucagon mRNA levels were significantly higher in rats fed rhubarb fiber than in those fed cellulose fiber (9.3 +/- 0.9 vs. 6.2 +/- 1.0 densitometer units). Proglucagon mRNA in the colon did not differ between diet treatments. Plasma c-peptide concentrations were significantly higher 30 min after an oral glucose tolerance test in the rhubarb vs. cellulose group (1627 +/- 67 vs. 1290 +/- 71 pmol/L). Passive permeability, measured by the uptake of L-glucose, was significantly higher in the jejunum of rats fed cellulose compared with those fed rhubarb fiber. Adjusting total glucose uptake for passive permeability and unstirred water layer resistance resulted in a higher Km being calculated for the jejunum and ileum of the cellulose fiber group. Jejunal and ileal carrier-mediated uptakes (Vmax) were not altered by diet and reflected the lack of difference between groups in sodium-dependent glucose cotransporter (SGLT-1) and sodium-independent glucose transporter (GLUT2) mRNA levels. Replacing cellulose fiber with rhubarb fiber in a diet upregulated ileal proglucagon mRNA and resulted in a reduced passive permeability but did not affect glucose transport of the small intestine. This work establishes the importance of dietary fiber fermentability in modulating intestinal proglucagon expression and possibly glucose homeostasis.

Ontogenic changes in proglucagon mRNA in BB diabetes prone and normal rats weaned onto a chow diet.

Weaning onto chow diets causes the highest incidence of diabetes in the BB rat. Changes in gut development and absorption of nutrients in the diabetes prone rat and the subsequent effect on pancreatic function may play a role in the ultimate development of the disease. BB diabetes prone (dp) and BB normal (n) dams were fed chow diets. Pups were killed at various ages ranging from 7 to 30 days. BBdp rats had higher small intestine and colon weights expressed per body weight at all ages (p < 0.0001). RNA content (mg/g) in the jejunum, ileum and colon was higher in the BBdp rats beginning at the critical period at 21 days and maintained at 24 days and 30 days (p < 0.0001). Proglucagon message decreased with age in both BBdp and BBn animals (p < 0.0001). Levels of proglucagon mRNA were higher in BBdp compared to BBn animals only in the ileum at 10 days (p < 0.01). Adjusting for total ileal and colonic RNA content resulted in BBdp animals having higher total colonic proglucagon mRNA at 21, 24 and 30 days (p < 0.0001). Plasma GLP-1(7-36) amide was more than doubled in BBdp compared to BBn animals (p < 0.0005) at 30 days. Expressing sodium-dependent D-glucose co-transporter (SGLT-1), GLUT2 and GLUT5 mRNA per total jejunal RNA shows increased transporter mRNA in BBdp compared to BBn rats at weaning (21 days) (p < 0.05). Radical differences exist between BBdp and BBn animals at 'critical periods' in both proglucagon and glucose transporter gene expression. These differences may help explain altered growth and diseases incidence between these two strains.