Polyphenols, metal ion complexation and biological consequences.
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Flavonoids↗
Biomedical subjects
D Tomé
Publications and source records attributed to D Tomé.
Effects of a high soy protein diet on intestinal polyamines and ornithine decarboxylase activity in rats.
This study was performed to determine whether intestinal luminal polyamine concentrations are affected by a high soy protein diet when compared with a high casein diet or a normoprotein casein diet. We also determined the effects of these diets, with differences in polyamines content, on mucosal polyamines and ornithine decarboxylase (ODC) activity to assess cell proliferation. Three groups of eight male Wistar rats were fed either a 50% soy protein diet, a 50% casein diet, or an 18% casein diet as a control. After 4 weeks of feeding, both intestinal content and mucosa were recovered. Polyamines were assayed by high performance liquid chromatography. ODC activity was measured by the release of (14)CO(2) from (14)C-L-ornithine. Luminal putrescine and cadaverine concentrations were higher in the jejunum than in the ileum, suggesting an absorptive process. The highest concentrations of intestinal polyamines were observed in rats fed the soy protein diet (P < 0.05). Only minor differences were observed in mucosal polyamines according to the diets. ODC activity was also higher in the intestinal mucosa of rats fed the high soy protein diet (P < 0.05). These results suggest that intestinal luminal polyamine concentrations and ODC activity are modulated by the dietary protein source.
Journal Article↗
Nutritional value of [15N]-soy protein isolate assessed from ileal digestibility and postprandial protein utilization in humans.
The purpose of this work was to assess the true oro-ileal digestibility, and to concurrently quantify the deamination of absorbed dietary nitrogen to examine the postprandial nutritional value of a soy protein isolate (SPI) in humans. To assess bioavailability and bioutilization of SPI, 10 healthy volunteers ingested 30 g of SPI, intrinsically and uniformly [15N]-labeled, added with 100 g of sucrose and water up to a final volume of 500 mL. True ileal digestibility was assessed by the [15N]-dilution method for 8 h by means of a naso-intestinal intubation technique. To describe and quantify exogenous nitrogen deamination for the same time period, urine and plasma samples were collected. True oro-ileal digestibility of SPI nitrogen was 91%. The amount of absorbed SPI amino acids used for nonoxidative disposal, i.e., postprandial biological value, was 86% 8 h after meal ingestion. Hence, net postprandial protein utilization of SPI was 78%. Compared to previous data that were assessed under the same condition in humans, the nutritional value of SPI is 92% of that in milk protein concentrate.
Adult↗
Net postprandial utilization of [15N]-labeled milk protein nitrogen is influenced by diet composition in humans.
The aim of this study was to follow the fate of dietary nitrogen to assess the postprandial utilization of purified milk protein and to determine the acute influence of energy nutrients. For this purpose, a [15N]-labeling dietary protein approach was used. Twenty-five subjects swallowed an ileal tube and ingested [15 N]-milk protein alone or supplemented with either milk fat or sucrose. The absorption and postprandial deamination of dietary protein was monitored for 8 h. Sucrose delayed the absorption of protein longer than fat, but the ileal digestibility did not differ among groups (94.5-94.8%). Sucrose, but not fat, significantly reduced the postprandial transfer of [15N]-milk nitrogen to urea. Consequently, the net postprandial protein utilization (NPPU) of milk protein calculated 8 h after meal ingestion was 80% when ingested either alone or supplemented with fat and was significantly greater with sucrose (NPPU = 85%). This study shows that energy nutrients do not affect the nitrogen absorption but modify the metabolic utilization of dietary protein in the phase of nitrogen gain. Our method provides information concerning the deamination kinetics of dietary amino acids and further allows the detection of differences of dietary protein utilization in acute conditions. The diet composition should be carefully considered, and protein quality must be determined under optimal conditions of utilization.
Adult↗
Assessment of net postprandial protein utilization of 15N-labelled milk nitrogen in human subjects.
The nutritional quality of milk proteins, evaluated both in terms of digestibility and postprandial oxidation and retention in human subjects, was investigated in this study. Five healthy adult volunteers were given 480 ml 15N-labelled milk (i.e. 190 mmol N). 15N was subsequently determined at the ileal level, using a naso-intestinal intubation technique, as well as at the faecal level. Plasma and urine were sampled for 8 h after meal ingestion. Dietary exogenous N recovered at the terminal ileum after 8 h reached 8.6 (SE 0.8) mmol while the amount collected in the faeces was 6.5 (SE 0.7) mmol after 5 d. The true ileal and faecal digestibilities were 95.5 (SE 0.4)% and 96.6 (SE 0.4)% respectively. The appearance of [15N]amino acids in the plasma was rapid and prolonged. The measurement of 15N in the body urea pool and in the N excreted in the urine allowed us to calculate the deamination occurring after [15N]milk protein absorption. The net postprandial protein utilization (i.e. NPPU = (Nabsorbed-Ndeaminated)/Ningested), calculated as an index of protein quality 8 h after milk ingestion, was 81.0 (SE 1.9)%. Our data confirm that milk protein has a high oro-ileal digestibility in man and demonstrate that milk protein has a high NPPU, an index corresponding to a period in which the dietary protein retention is maximal.
Adult↗
Bovine lactoferrin induces both mucosal and systemic immune response in mice.
Lactoferrin (Lf) is a milk iron-binding glycoprotein that plays a role in iron transport and acts as both a bacteriostatic and a growth modulating agent. The aim of this study was to investigate the nature of immune responses induced by repeated oral administration of bovine milk Lf in mice. Groups of ten female BALB/c mice were fed daily for 4 weeks with two doses of protein antigen: a low (0.05 mg/g body weight per d) or high (1 mg/g body weight per d) dose of Lf, or water as a control. A fourth group was immunized intramuscularly with 0.01 mg Lf in complete Freund's adjuvant. Anti-Lf IgA and IgG were detected in the intestinal fluid and serum of mice given Lf. Total immunoglobulins were higher in the intestinal fluid in Lf groups than in the control group. No difference could be detected in the serum. IgA and IgG secretion was enhanced in Peyer's patches and spleen from Lf-fed mice, in comparison with controls. [3H]thymidine uptake into Peyer's patch and spleen cells from both control and Lf-fed mice was enhanced by 75 micrograms Lf/ml in vitro, but Lf groups had a greater proliferation rate than the control group. These findings suggested that Lf could act as an immunostimulating factor on the mucosal immune system and that activation of the mucosal immune system is dependent on the ability of Lf to bind to the intestinal mucosa.
Administration, Oral↗
Secretion of sparfloxacin from the human intestinal Caco-2 cell line is altered by P-glycoprotein inhibitors.
The mechanism of intestinal secretion of the difluorinated quinolone sparfloxacin was investigated with the epithelial cell line Caco-2 and was compared to that of the P-glycoprotein (P-gp) substrate vinblastine. The P-gp inhibitors verapamil and progesterone significantly increased the epithelial cell accumulation of both vinblastine and sparfloxacin. This increase is likely to result from an inhibition of drug secretion since both vinblastine uptake and sparfloxacin uptake are known to proceed through a passive transmembrane diffusion. The unidirectional fluxes across cell monlayers grown on permeable filters indicated that a net secretion of sparfloxacin and vinblastine occurred across Caco-2 cells. These secretions were significantly inhibited by the MDR-reversing agent verapamil. We conclude that the P-gp is likely to be involved in the intestinal elimination of the difluorinated quinolone sparfloxacin.
ATP Binding Cassette Transporter, Subfamily B, Mem↗
Sodium-dependent and -independent transport of L-glutamate in the rat intestinal crypt-like cell line IEC-17.
Mechanisms of L-glutamate transport in intestinal crypts were investigated using the rat intestinal crypt-like cell line IEC-17. Kinetic analysis and competition experiments run in the presence or in the absence of extracellular sodium indicate that L-glutamate uptake occurs through three different transport components: (1) a high affinity Na+-independent component also carrying cystine, similar to system x(c)-; (2) a high affinity Na+-dependent component inhibited by D- and L-aspartate corresponding to the ubiquitous system X(A,G)-; and (3) a low affinity Na+-dependent system resembling the neutral amino acid transport system ASC. The simultaneous presence of these three components suggest that crypt cells are ready to face potential high variations of L-glutamate concentration in the intestinal villus environment.
Animals↗
Evidence for a passive diffusion mechanism for sparfloxacin uptake at the brush-border membrane of the human intestinal cell-line Caco-2.
The oral uptake of the new fluoroquinolone sparfloxacin was evaluated in the human epithelial cell line Caco-2 that possesses intestinal enterocyte-like properties when cultured in vitro. The uptake of [14C]-sparfloxacin across the apical membrane of Caco-2 cell monolayers was rapid and similar at 25 and 37 degrees C. The initial rate of sparfloxacin uptake was not saturable in the 1-200 microM range and was unaffected by metabolic inhibitors (depletion of ATP store or ouabain), indicating that uptake was energy-independent. The absence of competition with other fluoroquinolones or aminocephalosporins showed that the absorption of sparfloxacin did not involved the H+-coupled dipeptide transport system. Our findings suggest that the apical uptake of sparfloxacin by Caco-2 cells mainly involves diffusion, a finding that is in agreement with the high lipophilicity of sparfloxacin. The intracellular-to-extracellular concentration ratio of approximately 14 after 60 min of incubation suggests the existence of important binding of sparfloxacin to cell components.
Anti-Infective Agents↗
Whole-body protein turnover in humans fed a soy protein-rich vegetable diet.
OBJECTIVES: This study was designed to compare the whole-body protein turnover in humans after the ingestion of a soy protein-rich vegetable diet with that of a control group fed a western animal protein-rich diet. SUBJECTS: Twelve male volunteers were divided into two groups of six subjects who were given for two weeks either a 85% vegetable protein diet (diet VP) or a control western animal protein-rich diet (diet AP). INTERVENTIONS: Whole-body protein turnover was estimated at the end of the two-week controlled diet period using the [15N]-glycine end-product method. Nitrogen flux rates were determined in the fed state (1.3 g protein/kg) over a 9 h period after the dose of [15N]-glycine was given. RESULTS: After the 9 h of the test, the urinary ammonia excretion was significantly higher in the group receiving the diet AP than that in the group receiving the diet VP (P < 0.05). In contrast, there was no significant difference for both total nitrogen and urea nitrogen excretions. Both the protein synthesis and the protein breakdown were similar in both groups. In the same way, the net protein deposition measured in the fed state during 9 h was similar for both diets at 0.07 g/kg/h. CONCLUSIONS: Young adults fed 1.3 g/kg/d of either meat or vegetable protein-rich diet for two weeks did not show a different protein turnover.
Adult↗
Cryptosporidiosis-induced impairment of ion transport and Na+-glucose absorption in adult immunocompromised mice.
Electrolyte transport was investigated during chronic cryptosporidiosis in adult anti-interferon-gamma-treated SCID mice by means of Ussing chamber techniques. In basal conditions, infection of immunocompromised mice with Cryptosporidium parvum resulted in a 30% reduction (P < .05) in the ileal short-circuit (Isc) current related to a 28% reduction (P < .05) in tissue conductance compared with controls. The rises in Isc and transepithelial potential difference induced by glucose (10 mM) were significantly reduced by Cryptosporidium infection (P < .01) compared with controls. In contrast, responses to mucosal glutamine were marginally affected. Electrical parameters of the ileum were not affected by the addition of indomethacin or furosemide, in either control or Cryptosporidium-infected mice. Thus, long-term cryptosporidiosis in immunocompromised animals leads to a reduction in net ion exchanges, decreased paracellular shunting, and impaired Na+-glucose cotransport in the ileum, without prostanoid- or enterotoxin-mediated electrogenic Cl- secretion.
Animals↗
Fasting and postprandial polyamine concentrations in the human digestive lumen.
Polyamines are essential to cellular proliferation and differentiation. The gastrointestinal tract could represent a major source of polyamines in the body; however, there is little information regarding the presence of polyamines in the human intestinal chyme, and the source of these intraluminal polyamines remains unclear. The aims of our study were to determine the concentrations and flow rates of polyamines in the human intestinal lumen and to estimate the contribution from food to these concentrations. Polyamine concentrations and flow rates were determined after 12 h of fasting in jejunal (n = 25) and ileal (n = 9) effluents collected by the slow-marker perfusion technique. Kinetic studies were performed after water ingestion (no polyamines) in the jejunum (n = 6) and ileum (n = 5) and in the jejunum after a yogurt test meal (polyamine content: 2.8 mumol putrescine, 2.1 mumol cadaverine, 2.1 mumol spermidine, and 1.9 mumol spermine; n = 9). There were significant polyamine concentrations in the lumen of the human gut during the fasting state, suggesting endogenous secretion. Higher polyamine concentrations were observed in the jejunum than in the ileum (P < 0.05), suggesting proximal absorption. Kinetic studies showed a 25% transitory increase in the jejunal putrescine flow rate after ingestion of the yogurt test meal, suggesting that dietary polyamines are fully absorbed.
Adult↗
[15N]-labeled pea flour protein nitrogen exhibits good ileal digestibility and postprandial retention in humans.
The aim of the present study was to evaluate postprandial absorption of pea protein as well as exogenous nitrogen retention in humans. For this purpose, after fasting overnight, seven healthy adults (4 males and 3 females) ingested [15N]-labeled pea protein (195 mmol N). Ileal effluents were collected for 8 h at 30-min intervals using a nasointestinal intubation technique. Urine and plasma samples were collected for 24 h. The [15N]-enrichment was determined in the intestinal samples, in the plasma amino acids and urea as well as in the urinary urea and ammonia fractions. The true gastroileal absorption of pea protein was 89.4 +/- 1.1%. This absorption was correlated with a significant increase (P < 0.05) in [15N]-enrichment in the plasma amino acids and in the nitrogen incorporated into the body urea pool for 1 h following pea ingestion. The enrichment remained significantly higher than the basal values in these pools 24 h after pea ingestion. The recovery of total urinary exogenous nitrogen after 22 h was 31.1 +/- 9.3 mmol N. Moreover, the kinetics of [15N]-labeled pea amino acids deamination reached a plateau of 39 mmol. Under these conditions, pea nitrogen retention represented 78% of the absorbed dietary nitrogen in healthy humans. The present results demonstrate the good true nitrogen digestibility and retention of pea protein in humans.
Adult↗
The inhibition of intestinal dipeptidylaminopeptidase-IV promotes the absorption of enterostatin and des-arginine-enterostatin across rat jejunum in vitro.
Transport of enterostatin (VPDPR) across rat jejunum was investigated using Grass-Sweetana diffusion chambers. The rate of absorption of enterostatin and its metabolites were studied in absence and in presence of diisopropylfluorophosphate (DFP), a serine protease inhibitor. An extensive hydrolysis of enterostatin was observed during incubation with brush border membranes and when exposed to the mucosal side of the jejunal epithelium. No accumulation of enterostatin occurred in serosal tissue. Addition of DFP delayed enterostatin disappearance and abolished des-arg-enterostatin degradation. Under these conditions, a low amount of enterostatin was able to cross the epithelium intact. Moreover, a substantial amount of des-arg-enterostatin is absorbed across the jejunal epithelium, probably through passive diffusion. Thus, a decreased metabolic activity increased the absorption of a tetrapeptide (VPDP). Dipeptidylaminopeptidase IV, remains a limiting step in transfer of intact enterostatin and its metabolite des-arg-enterostatin across intestinal wall.
Animals↗
Gastrojejunal kinetics and the digestion of [15N]beta-lactoglobulin and casein in humans: the influence of the nature and quantity of the protein.
The evolution and luminal effects of different quantities of casein and beta-lactoglobulin were investigated in the upper jejunum of 35 volunteers who ingested 400 mL water with either beta-lactoglobulin or casein in either low or high doses (72.6 mmol N, Lbetalg; 71.7 mmol N, LCas; 368.2 mmol N, Hbetalg; 386.8 mmol N, HCas). The flow rate of the liquid effluents as well as the nitrogen movements were measured and the exogenous ([15N]) and endogenous nitrogen fractions analyzed in the upper jejunum after milk protein ingestion. The basal jejunal liquid flow rate (mL/min) was 3.88+/-1.84 and peaked in the 0-20 min period for water (9.92+/-3.72) and Lbetalg (7.27+/-3.08) and during the 20-40 min period for LCas (5.69+/-2.49), HCas (6.32+/-1.85), and Hbetalg (6.11+/-2.31). One hour after water, LCas, Lbetalg, Hbetalg, and HCas ingestion, 100%, 95%, 85%, 71%, but only 38% of the liquid phase of the meal were passed through the jejunum, respectively. The flow rate of the endogenous nitrogen was 12.93+/-5.22 mmol N/h before meal ingestion; remained unchanged after water, LCas or Hbetalg ingestion; but increased significantly (P<0.05) after Lbetalg and HCas ingestion. The net disappearance of exogenous nitrogen in the upper jejunum 240 min after HCas, Lbetalg, LCas and Hbetalg ingestion was 82.6+/-9.5%, 61.6+/-9.6%, 58.4+/-14.7%, and 44.7%+/-24.4%, respectively. The exogenous fraction of protein nitrogen recovered in the upper intestinal lumen represented 43.3% of the ingested Hbetalg nitrogen, but only 4.9% of the ingested HCas nitrogen. In conclusion, casein and beta-lactoglobulin present differences in both the intestinal kinetics of amino acid delivery and in the nature of the products in the intestinal lumen. These differences have to be taken into account from both nutritional and physiologic points of view for the utilization of these proteins in humans.
Adult↗
Modulation of putrescine transport in rat intestinal brush-border membrane vesicles by fasting and refeeding.
Fasting and refeeding dramatically alter small intestinal mucosal growth which is greatly dependent on polyamine biosynthesis and transport. The aim of this study was therefore to examine the uptake of the diamine putrescine by brush-border membrane vesicles from the small intestine of rats fasted for 3 days or refed a standard diet after a PERIOD OF FASTING. WHILE THE MICHAELIS-MENTEN CONSTANT KM WAS essentially unaltered, the maximum velocity (Vmax) for putrescine uptake was 1.85-fold higher in fasted animals than in ad libitum-fed controls. Refeeding fasted rats for 24 h caused a 31% decrease in the Vmax value that, however, remained 1.27-fold higher than in control RATS, WHILE THE KM VALUE WAS STILL UNCHANGED. FASTING RATS OR refeeding rats after a period of fasting caused only a 13 or 17% increase, respectively, in the value of the constant for the nonsaturable component (P) of putrescine transport relative to the corresponding control condition. Our study also confirms that both the mucosal polyamine biosynthesis and intestinal content are altered by fasting. We suggest that an increased uptake activity may have a conservative role by preventing a substantial loss of tissue polyamines during fasting.
Animals↗
A 15N-leucine-dilution method to measure endogenous contribution to luminal nitrogen in the human upper jejunum.
OBJECTIVE: This study was done to investigate whether an intraveinous infusion of 15N-leucine was accurate to differentiate between endogenous and exogenous nitrogen in the human jejunum after meal ingestion. SUBJECTS: Four healthy human volunteers equipped with an upper jejunal tube. INTERVENTIONS: The jejunal effluents were collected both under fasting conditions and after ingestion of 300g of yoghurt. The nitrogen, amino acid composition and 15N-leucine enrichment were determined in the digesta. RESULTS: During fasting, the jejunal flow rates (mmol/h) of both total nitrogen and amino acids were stable (6.9 +/- 2.7 and 1.88 +/- 0.79, respectively). After yoghurt ingestion, the flow rate of total nitrogen increased to 28.6 +/- 5.8mmol/h at 2h. The 15N-leucine enrichment in plasma reached a plateau at 4.3 mole % excess after one hour and did not vary significantly after meal ingestion. The 15N-leucine enrichment of the endogenous secretion (Ee) in the jejunum was fitted by the equation: Ee = 2.18[1 - 2.05 x exp( - 0.42 x t)]. After yoghurt ingestion, the enrichment in jejunal secretions decreased during the first 80 min. The endogenous nitrogen, calculated from the 15N-enrichment, significantly increased from 20 to 40min after meal ingestion compared to the basal value (P < 0.05). The estimation of the exogenous nitrogen and amino acid yield 300min after yoghurt ingestion indicated that 62 +/- 30% of the exogenous nitrogen and 75 +/- 12% of the amino acids were absorbed in the upper jejunum. CONCLUSIONS: The 15N-leucine-dilution method appears to be a convenient method to differentiate between the exogenous and endogenous contributions to nitrogen fluxes in the intestinal digesta of humans. It can be used in association with dietary protein labelling or in substitution when no labelled dietary proteins are available to compare the digestion as well as the absorption of meals at different levels of the intestine.
Adult↗
Putrescine transport in differentiated Caco-2 cells.
OBJECTIVE AND METHODS: To evaluate the putative importance of digestive luminal polyamines, we investigated the role of oxidation in putrescine uptake and transport in differentiated Caco-2 cells grown in monolayer. RESULTS: Our results confirm a putrescine uptake system located both at the apical and basolateral cell membrane. Putrescine is efficiently transported and excreted in the apical-to-basolateral direction. CONCLUSION: These results suggest (i) a diamine oxidase controlled catabolic oxidative excreting pathway and (ii) a paracellular route for putrescine transport. This catabolic route seems to proceed by converting putrescine into gamma-aminobutyraldehyde which is then rapidly excreted from the cell at the basolateral membrane since no gamma-aminobutryraldehyde was detected in the cell. This putative basolateral excreting mechanism is inhibited in the presence of the diamine oxidase inhibitor, aminoguanidine.
Biological Transport, Active↗