Contribution of intestinal microbial lysine to lysine homeostasis is reduced in minipigs fed a wheat gluten-based diet

BACKGROUND: We previously reported microbial lysine contribution to plasma lysine homeostasis in humans with an adequate lysine intake. OBJECTIVE: We sought to explore whether the low lysine intake from a wheat gluten-based diet is balanced by enhanced microbial lysine contribution in a pig model. DESIGN: Twenty miniature pigs (minipigs) fitted with ileo-ileal cannulas were fed 2 wheat gluten-based diets. One diet provided 2.7 g lysine/kg diet (WG diet) and one diet was supplemented with crystalline lysine to provide 6.6 g lysine/kg diet (WG+Lys diet). Both diets were fed for 10 or 100 d (n = 5 per group): 10WG+Lys, 10WG, 100WG+Lys, and 100WG diets. Ileal microbial lysine, which we considered to be the precursor pool for absorption, was labeled by oral administration of (15)NH(4)Cl for the final 10 d. On days 10 and 100, a 10-h fast-fed tracer protocol with [1-(13)C]lysine was performed. RESULTS: Lysine rates of appearance decreased by 25% with the WG diet in the fed state but increased by 50% with the WG+Lys diet in the fasted state (P < 0.05). Daily gross microbial lysine contribution was lower (P < 0.05) with the WG diet (205.3 micro mol. kg(-) (1). d(-)(1)) than with the WG+Lys diet (370.7 micro mol. kg(-) (1). d(-)(1)), irrespective of the adaptation period and was similar to the ileal lysine loss with the WG diet. In the WG groups, incorporation of microbial lysine increased in the duodenum and liver (P < 0.05) but not in whole-body and muscle proteins. CONCLUSION: Minipigs fed the WG diet did not adapt by showing an enhanced absorption of microbial lysine to the extrasplanchnic tissues, presumably because microbial lysine continues to be used for splanchnic protein synthesis.     

Plasma proteins in growing analbuminaemic rats fed on a diet of low-protein content

1. Analbuminaemic and Sprague-Dawley (control) rats were fed on low- (60 g/kg) protein and control (200 g protein/kg) diets ad lib. from weaning. Males and females were studied separately. Body-weight and plasma protein concentrations were determined at 10 d intervals from 25 to 75 d of age. Electrophoresis of plasma proteins was performed in samples from day 75. Extracellular fluid volume was measured at 10 d intervals from day 45 onwards. Colloid osmotic pressure was measured in plasma and interstitial fluid (wick technique) at the start and end of the trial. 2. Body-weight increased much less on the low-protein diet than on the normal diet in both strains and sexes. The growth retardation was slightly more pronounced in the male analbuminaemic rats than in the male Sprague-Dawley controls. 3. Plasma protein concentration increased during normal growth in all groups, particularly in the female analbuminaemic rats. This increase was reduced by the 60 g protein/kg diet in all groups, with the exception of the male analbuminaemic rats. 4. Differences in plasma colloid osmotic pressure were similar to those seen in plasma protein concentration. Interstitial colloid osmotic pressure was higher in the control rats than in the analbuminaemic ones. The interstitial colloid osmotic pressure increased during growth in the control but not in the analbuminaemic rats. The difference in interstitial colloid osmotic pressure between the strains was maintained during low-protein intake, but at a lower level than during normal protein intake. 5. Subtracting interstitial from plasma colloid osmotic pressure, resulted in a rather similar transcapillary oncotic gradient in the various groups at 75 d, both on the control protein diet (11-14 mmHg), and on the low-protein diet (9-11 mmHg). 6. All protein fractions were reduced to a similar extent by the low-protein diet in the control rats, whereas in the analbuminaemic rats protein fractions produced in the liver were more severely depressed. 7. Extracellular fluid volume as a percentage of body-weight was similar in all groups, and decreased with increasing age. 8. In conclusion, the analbuminaemic rats were able to maintain the transcapillary oncotic gradient on both diets by reducing the interstitial colloid osmotic pressure. Oedema was not observed. 9. Despite the absence of albumin, the protein-malnourished analbuminaemic rat is no more susceptible to hypoproteinaemia and oedema than its normal counterpart.     

Effects of sn-2 palmitic acid-fortified vegetable oil and fructooligosaccharide on calcium metabolism in growing rats fed casein based diet

This study was carried out to investigate the efficacy of sn-2 palmitic acid-fortified vegetable oil (Sn2PA) on calcium absorption and to confirm the synergistic effects of fructooligosaccharide on calcium absorption. Male SD rats were fed 6 kinds of casein based diets containing vegetable oil (control), sn-2 palmitic acid-fortified vegetable oil (Sn2PA) and Sn2PA with fructooligosaccharide(Sn2PAFO) in two levels of calcium (normal 0.5% and high 1.0%) for 3 weeks. Total lipids, cholesterol, triglyceride and calcium in blood were measured. Feces were collected using cages for 4 days. Serum concentrations of total lipids and calcium were not significantly different among groups. However, serum triglyceride was significantly decreased by fructooligosaccharide supplementation regardless of dietary calcium level. The lipid absorption was not significantly different among experimental groups. Calcium absorption was significantly higher in Sn2PAFO group than other groups. Calcium solubility of intestine was increased by sn-2 palmitic acid supplementation. These results suggest that sn-2 palmitic acid and fructooligosaccharide supplementation could be beneficial for baby foods including infant formula, with regard to increasing absorption of calcium by more soluble calcium in the small intestinal content.     

In vivo intestinal calcium transport in infant rats: normal and growth retarded.

We compared the in vivo transport of calcium (Ca) in the jejunum + ileum of 1-, 2-, 3-, 4-, and 6-week old rats. In normal rats, net absorption and the estimated bidirectional fluxes of Ca (lumen-to mucosa and mucosa-to-lumen) normalized for differences in size of the intestine (mumoles/hour per g weight) were similar in the 1-, 2-, and 3-week old rats and significantly lower in the 6-week old than in the younger rats. Growth retardation (pups raised with mothers fed a diet deficient in protein) appeared to have suppressed net absorption and the bidirectional fluxes of calcium in the 4- and 6-week old rats, but to have slightly enhanced net absorption and the bidirectional fluxes in the 1-, 2-, and 3-week old rats. These findings suggested a change in the mechanism(s) for intestinal transport of Ca during maturation. Rate of gain in body weight expressed as g/day increased with age from 1.4 g/day between 1 and 2 weeks, to 6.1 g/day between 4 and 6 weeks of age. There was no correlation between rate of net Ca absorption from the jejunum + ileum and gain in body weight expressed as g/day or as percent per day.     

Stimulation of postprandial in vivo glycogenesis and lipogenesis of rats fed high fructose diet with varied phosphate content

It has been reported that increased fructose intake is associated with the development of the metabolic syndrome. The phosphate (P) sequestering capacity of fructose is likely to affect the phosphorylation capacity of different metabolites, and this, in turn, may be the basis for several metabolic derangements, especially in the P requiring reactions, for example, glycogenesis and lipogenesis. We hypothesized that P enrichment of the diet can balance P status and, consequently, affect glycogenesis and lipogenesis. An animal experiment was executed in which adult male Sprague-Dawley rats were maintained for 4 days on high-fructose diets with different P content (0.15%, 0.165%, 0.30%, and 1.65%). At the end of the feeding period, overnight fasted rats were tube fed a test meal, injected with ³H₂O and euthanized 1 hour later. Final plasma glucose, insulin, uric acid, and triacylglycerol concentrations, as well as in vivo rates of glycogen and lipid synthesis and hepatic glycogen content, were measured. Results showed that increased P content of the diet was associated with an increase in postprandial epididymal fat pad (P = .007) and hepatic lipogenesis (P = .029), as well as glycogenesis (P = .024). In conclusion, P content of the diet was found to stimulate both glycogenesis and lipogenesis. These alterations in carbohydrate and fat metabolism point to the potential of P in influencing nutritional status.     

Relationship between amino acid composition of diet and plasma cholesterol level in growing rats fed a high cholesterol diet

The effects of dietary sulfur-containing amino acids and glycine on plasma cholesterol level were studied in rats fed amino acid mixture diets containing cholesterol. The relationship between the amino acid composition of dietary proteins and plasma cholesterol levels was also investigated in rats fed diets containing various kinds of protein such as casein, egg albumin, pork protein, fish protein, corn gluten, wheat gluten and soy protein. Feeding the amino acid mixture corresponding to casein led to an approximately two-fold level of plasma total cholesterol as compared with feeding the amino acid mixture corresponding to wheat gluten. It was possible to reduce the plasma cholesterol of rats fed the amino acid mixture of the casein type by increasing the proportion of cystine in the total sulfur amino acids. Inversely, the deprivation of cystine resulted in an enhancement of the plasma cholesterol of rats fed the gluten type amino acid mixture. Glycine had a tendency to resist increases in the plasma cholesterol level. A significant negative correlation was noted between plasma cholesterol levels and the content of cystine in intact dietary proteins. The results suggest that the differential effect of dietary proteins on plasma cholesterol level is mainly associated with sulfur-containing amino acids included in the protein, regardless of whether it is of animal or plant origin.     

Effect of colostomy on nitrogen nutrition in the chicken fed a low protein diet plus urea

Recent interest in the role of the ceca in avian nutrition has focused on recycling of urinary nitrogen through the ceca. In the colostomized chicken, we observed appreciable decrease in utilization of dietary urea nitrogen. This is the first report demonstrating that the ceca are able to play an advantageous role in nitrogen nutrition of the chicken, because back-flow of urinary nitrogen into the ceca was completely inhibited by colostomy. When colostomized chickens were fed a diet containing urea, little urea was found in feces but the amount of urea excreted in the urine corresponded to 77.5% of urea intake. Droppings of normal control chickens fed the same diet did not contain urea. However, they contained twice as much ammonia as the urine plus feces of colostomized chickens, indicating active and great degradation of urinary urea to ammonia by microflora in the ceca of control chickens. The recycling of urinary nitrogen through the ceca may be involved in the utilization of dietary urea by the chicken.     

Inhibition of intestinal calcium uptake by cadmium and the effect of a low calcium diet on cadmium retention

The direct effect of cadium (Cd) on the in vitro uptake of calcium by rat duodenum was examined by incubating mucosal tissue for 1 to 5 min in an isotonic medium containing ⁴⁵Ca with [³H] inulin included to correct for calcium (Ca) uptake associated with the space accessible to inulin. Cadium inhibited a second phase, time-dependent uptake of Ca which is influenced by vitamin D status and dietary levels of calcium. Cadium had no effect on either an initial rapid uptake of Ca (this is complete within one min) or the inulin space of the intestinal mucosa. Kinetic analysis of the Cd inhibition using 1.0 and 3.0 mm Ca against 0.0–1.0 mm Cd demonstrated Cd was a noncompetitive inhibitor with a K_i of 0.8 mm. Alternatively, Ca did not inhibit ^115mCd uptake in animals fed a normal diet. Furthermore, in animals fed a diet low in Ca for seven days, Cd uptake was not affected despite an increased Ca uptake.Whole-body retention and distribution studies of ^115mCd demonstrated that young rats pair-fed a low Ca diet for seven days retained more ^115mCd for nine days from a single oral dose of 100 nmol of CdCl₂ than control animals fed a Ca replete diet. The low Ca diet influenced organ distribution causing more Cd to be retained in the small intestine and kidneys. However, there was no significant change in the sum of the total Cd content in the kidney and liver which are the major sequestering organs of absorbed Cd.     

Effects of dietary calcium on atherosclerosis, aortic calcification, and icterus in rabbits fed a supplemental cholesterol diet

Background  

Vascular calcification is implicated in myocardial infarction, instability and rigidity of the aortic wall, and bioprosthetic failures. Although an increase in the calcium (Ca) content in atherogenic diets has been shown to decrease atherosclerosis in rabbits, whether Ca supplementation and deficiency can affect atherosclerosis-related aortic calcification remains unknown.     

Lipid absorption and intestinal tumour incidence in rats fed on varying levels of calcium and butterfat

The purpose of the 2 x 2 factorial study was to determine the effect of varying levels of dietary calcium (2.5 and 10 g/kg) and butterfat (50 and 200 g/kg) on lipid utilization and on development of colon tumours in animals initiated with 1,2-dimethylhydrazine dihydrochloride. Among rats fed on 200 g butterfat/kg, the fourfold increase in Ca intake induced more than a sevenfold increase in faecal excretion of total lipids and almost a fortyfold increase in faecal excretion of acid-extractable lipid. Among rats fed on 50 g butterfat/kg, the ingestion of supplemental Ca had a less dramatic effect and induced only a twofold increase in faecal excretion of total lipids and a threefold increase in acid-extractable lipid. The volume of intestinal adenocarcinomas was correlated with the excretion of acid-extractable lipid in faeces (R 0.369, P less than 0.02). Caecal enzymic activity was not correlated with tumour incidence or size or faecal lipid excretion. Overall, the fourfold increase in Ca intakes decreased total lipid absorption significantly but by less than 6%.     

High-viscosity carboxymethylcellulose reduces carbachol-stimulated intestinal chloride secretion in weaned piglets fed a diet based on skimmed milk powder and maltodextrin

High-viscosity carboxymethylcellulose (CMC) promotes gastrointestinal disorders, tissue alterations and bacterial overgrowth in pigs. The impact of CMC on intestinal absorptive and secretory physiology is not known. We hypothesised that CMC consumption alters intestinal Na-dependent glucose absorption and stimulates electrogenic chloride secretion. For testing this hypothesis, twenty-four piglets were weaned at 21 d of age and pair-fed for 13 d a starter diet based on skimmed milk powder and maltodextrin containing cellulose (control) or CMC. Body weight and faecal total aerobe and coliform counts were measured kinetically. At slaughter, digesta were weighed and characterised for viscosity and pH. Gastrointestinal tissues were weighed and sampled for physiology in Ussing chambers, morphometry and enzymology. Glucose absorption tended to be higher (P = 0.08) and carbachol-stimulated chloride secretion was lower (P = 0.01) with CMC in the small intestine, without changes in the colon. Aerobes were transiently higher at day 7 (P < 0.05) but coliform counts remained unchanged (P = 0.78) and beta-haemolitic Escherichia coli were virtually absent. Stomach and small-intestinal segments were heavier, and viscosity higher with CMC (0.001 < P < 0.05). The pH in the stomach was higher, and in the caecum and proximal colon lower with CMC (0.001 < P < 0.05). Jejunal villus area was slightly reduced with CMC (P < 0.05) without effects on enzyme activities (P > 0.10). In conclusion, CMC supplementation had pro-absorptive effects on the small intestine, possibly due to the absence of pathogenic E. coli in the present study.     

Effects of taurine supplementation on bone mineral density in ovariectomized rats fed calcium deficient diet

Taurine supplementation has been shown to have a beneficial effect on femur bone mineral content in ovariectomized rats. It therefore seemed desirable to find out whether the beneficial effect of taurine on ovariectomized rats fed calcium deficient diet could also be reproduced. Forty female Sprague-Dawley rats were divided into two groups. One group was OVX and the other group received sham operation (SHAM), and received either control diet or a taurine supplemented diet for 6 weeks. All rats were fed on calcium deficient diet (AIN-93: 50% level of calcium) and deionized water. Bone mineral density (BMD) and bone mineral content (BMC) were measured in spine and femur. The serum and urine concentrations of calcium and phosphorus were determined. Bone formation was measured by serum osteocalcin and alkaline phosphatase (ALP) concentrations. Bone resorption rate was measured by deoxypyridinoline (DPD) crosslinks immunoassay and corrected for creatinine. Urinary calcium and phosphorus excretion, osteocalcin in blood and cross link value were not significantly different among the groups. Within the OVX group, the taurine supplemented group had not higher femur bone mineral content than the control group. This study established the need for a study on the taurine effect on bone with different calcium levels.     

Decreased insulin secretion in islets from rats fed a low protein diet is associated with a reduced PKAalpha expression

A low protein diet has been shown to affect the amount and activity of several enzymes and to decrease insulin secretion by islets isolated from rats fed such a diet. To understand the mechanisms involved in this phenomenon, we investigated the effects of forskolin, a stimulator of adenylyl cyclase, on insulin secretion by pancreatic islets from rats fed a normal (17%; NP) or low (6%; LP) protein diet for 8 wk. Isolated islets were incubated for 1 h in Krebs-bicarbonate solution containing 8.3 mmol glucose/L, with or without 10 micromol forskolin/L. The forskolin-induced insulin secretion was higher in islets from NP rats than in those from LP rats (P<0.05). Western blotting revealed that the amount of the alpha catalytic subunit of protein kinase A (PKAalpha) was 35% lower in islets from LP rats than in islets from NP rats (P<0.05). Moreover, PKAalpha mRNA expression was reduced by 30% in islets from LP rats (P<0.05). Our results indicated a possible relationship between a low protein diet and a reduction in PKAalpha expression. These alterations in PKAalpha may be responsible in part for the decreased insulin secretion by islets from rats fed a low protein diet.     

Dietary calcium phosphate stimulates intestinal lactobacilli and decreases the severity of a salmonella infection in rats

We have shown recently that dietary calcium phosphate (CaPi) has a trophic effect on the intestinal microflora and strongly protects against salmonella infection. It was speculated that precipitation by CaPi of intestinal surfactants, such as bile acids and fatty acids, reduced the cytotoxicity of intestinal contents and favored growth of the microflora. Because lactobacilli may have antagonistic activity against pathogens, the main purpose of the present study was to examine whether this CaPi-induced protection coincides with a reinforcement of the endogenous lactobacilli. In vitro, Salmonella enteritidis appeared to be insensitive to bile acids and fatty acids, whereas Lactobacillus acidophilus was killed by physiologically relevant concentrations of these surfactants. Additionally, after adaptation to a purified diet differing only in CaPi concentration (20 and 180 mmol CaHPO4. 2H2O/kg), rats (n = 8) were orally infected with S. enteritidis. Besides reducing the cytotoxicity and the concentration of bile acids and fatty acids of ileal contents and fecal water, CaPi notably changed the composition of ileal bile acids in a less cell-damaging direction. Significantly greater numbers of ileal and fecal lactobacilli were detected in noninfected, CaPi-supplemented rats. As judged by the lower urinary NOx excretion, which is a biomarker of intestinal bacterial translocation, dietary CaPi reduced the invasion of salmonella. Additionally, the colonization resistance was improved considering the reduction of excreted fecal salmonella. In accordance, fewer viable salmonella were detected in ileal contents and on the ileal mucosa in the CaPi group. In conclusion, reducing the intestinal surfactant concentration by dietary CaPi strengthens the endogenous lactobacilli and increases the resistance to salmonella.