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1.
Food Chem ; 249: 111-118, 2018 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-29407913

RESUMO

We studied the kinetics of peptide release during the gastric digestion of meat proteins in vivo, in view to predicting the release of bioactive peptides further on in the digestive tract. Six mini pigs fitted with gastric cannulas received a meal with cooked beef as protein source. Digesta was collected at regular time intervals up to 5½â€¯h. The peptides generated by the gastric digestion of meat were identified and quantified using label-free LC MS, thereafter subjected to in silico digestion mimicking the action of intestinal enzymes. Three clusters of proteins presenting similar evolutions according to their dynamic hydrolysis were obtained. This study clearly improves the in silico prediction of the intestinal release of bioactive peptides by mapping meat protein degradation in the stomach in an in vivo model. Knowledge of the conformation of the peptides released in the stomach further improves this prediction.


Assuntos
Carne/análise , Peptídeos/metabolismo , Suínos/metabolismo , Ração Animal/análise , Animais , Bovinos , Cromatografia Líquida , Simulação por Computador , Proteínas Alimentares/química , Proteínas Alimentares/metabolismo , Digestão , Mucosa Gástrica/metabolismo , Cinética , Mapeamento de Peptídeos , Peptídeos/química , Proteólise , Suínos/crescimento & desenvolvimento , Espectrometria de Massas em Tandem
2.
Br J Nutr ; 116(12): 2091-2096, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28069089

RESUMO

Lycopene (LYC) bioavailability is relatively low and highly variable, because of the influence of several factors. Recent in vitro data have suggested that dietary Ca can impair LYC micellarisation, but there is no evidence whether this can lead to decreased LYC absorption efficiency in humans. Our objective was to assess whether a nutritional dose of Ca impairs dietary LYC bioavailability and to study the mechanism(s) involved. First, in a randomised, two-way cross-over study, ten healthy adults consumed either a test meal that provided 19-mg (all-E)-LYC from tomato paste or the same meal plus 500-mg calcium carbonate as a supplement. Plasma LYC concentration was measured at regular time intervals over 7 h postprandially. In a second approach, an in vitro digestion model was used to assess the effect of increasing Ca doses on LYC micellarisation and on the size and zeta potential of the mixed micelles produced during digestion of a complex food matrix. LYC bioavailability was diminished by 83 % following the addition of Ca in the test meal. In vitro, Ca affected neither LYC micellarisation nor mixed micelle size but it decreased the absolute value of their charge by 39 %. In conclusion, a nutritional dose of Ca can impair dietary LYC bioavailability in healthy humans. This inhibition could be due to the fact that Ca diminishes the electrical charge of micelles. These results call for a thorough assessment of the effects of Ca, or other divalent minerals, on the bioavailability of other carotenoids and lipophilic micronutrients.


Assuntos
Cálcio da Dieta/efeitos adversos , Carotenoides/antagonistas & inibidores , Suplementos Nutricionais/efeitos adversos , Digestão , Frutas/química , Absorção Intestinal , Solanum lycopersicum/química , Adulto , Carbonato de Cálcio/administração & dosagem , Doenças Cardiovasculares/epidemiologia , Doenças Cardiovasculares/etiologia , Carotenoides/sangue , Carotenoides/metabolismo , Estudos Cross-Over , Feminino , França/epidemiologia , Humanos , Incidência , Licopeno , Masculino , Refeições , Micelas , Valor Nutritivo , Neoplasias da Próstata/epidemiologia , Neoplasias da Próstata/etiologia , Risco , Propriedades de Superfície , Adulto Jovem
3.
J Nutr Health Aging ; 18(9): 814-9, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25389959

RESUMO

OBJECTIVE: Glutamine is the preferred fuel for the rat small intestine and promotes the growth of intestinal mucosa, especially in the event of gut injury. Quantitatively, glutamine is one important precursor for intestinal citrulline release. The aim of this study was to determine whether the effect of glutamine on the increase in intestinal villus height is correlated with an increase in both gut mass and citrulline plasma level in very old rats. METHODS: We intermittently supplemented very old (27-mo) female rats with oral glutamine (20% of diet protein). Intestinal histomorphometric analysis of the small bowel was performed. Amino acids, in particular citrulline, were measured in the plasma, liver and jejunum. Markers of renal (creatinine, urea) and liver (alanine aminotransferase [ALT]) and aspartate aminotransferase (AST) functions were measured to evaluate renal and liver functions in relation to aging and to glutamine supplementation. Liver glutathione was also determined to evaluate cellular redox state. RESULTS: Glutamine supplementation maintains the body weight of very old rats, not by limiting sarcopenia but rather by increasing the organ mass of the splanchnic area. Total intestine mass was significantly higher in glutamine-supplemented rats than in controls (15%). Measurement of villus height and crypt depth demonstrated that the difference between villus and crypt was significantly improved in glutamine pre-treated rats compared to controls (~ 11%). Plasma citrulline also increased by 15% in glutamine-supplemented rats compared to controls. CONCLUSION: Citrulline appears as a biomarker of enterocyte mass in villous atrophy associated with advanced age. Non-invasive measurement of this metabolite may be useful in following the state of the gastrointestinal tract in very old people, whose numbers are increasing worldwide and the care of whom is a major public health issue. The gut may contribute to the malnutrition caused by malabsorption frequently observed in the elderly.


Assuntos
Envelhecimento/fisiologia , Citrulina/sangue , Glutamina/administração & dosagem , Intestino Delgado/anatomia & histologia , Intestino Delgado/efeitos dos fármacos , Aminoácidos/análise , Aminoácidos/sangue , Animais , Peso Corporal/efeitos dos fármacos , Suplementos Nutricionais , Esquema de Medicação , Feminino , Glutamina/análise , Glutamina/sangue , Glutationa/metabolismo , Mucosa Intestinal/anatomia & histologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Intestino Delgado/patologia , Intestino Delgado/fisiologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Ratos , Ratos Wistar , Fatores de Tempo
4.
J Dairy Sci ; 96(10): 6690-6, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23972496

RESUMO

Ruminal microbes have the capacity to inactivate ochratoxins, rendering ruminants less sensitive to this fungal contaminant found in cereal feeds. However, ochratoxin A has been reported in milk surveys. The objective of this study was to assess the toxicokinetics, excretion, and transmission into milk of ochratoxin A using doses similar to those of naturally occurring field contaminations. Six Lacaune dairy ewes in late lactation were separated into 2 groups that received a single dose of contaminated wheat containing 5 or 30 µg of ochratoxin A/kg of body weight. After administration, toxicokinetics and excretion were monitored for 48 h. Subsequently, ewes were administered the corresponding toxin dose daily for 24 d followed by a second toxicokinetics and excretion monitoring period for this long-term exposure. The doses used did not affect production or health of ewes. After a single dose, ochratoxin A and its main metabolite, ochratoxin α, were found in blood 1h postexposure. The maximum blood concentrations of ochratoxin A and α, respectively, were dose dependent and were observed, on average, 6 and 8h after exposure. Long-term exposure increased the maximum concentration of ochratoxin A detected in blood, whereas ochratoxin α was not affected. In contrast, the time to reach the maximum concentration was reduced to 3h for both molecules. Ochratoxins, essentially ochratoxin α, were mainly excreted in feces. Ochratoxin A and α were detected in milk at concentrations that were dose dependent but with a low carryover rate (<0.02%). Chronic administration did not increase the concentration of toxin in milk. Even though ochratoxin A can escape ruminal degradation and traces were found in milk of experimentally exposed ewes, the low carryover of ochratoxin A in milk minimizes the risk to consumers.


Assuntos
Contaminação de Alimentos , Leite/química , Ocratoxinas/análise , Ocratoxinas/farmacocinética , Ovinos/fisiologia , Triticum/toxicidade , Animais , Peso Corporal , Indústria de Laticínios , Ingestão de Alimentos , Fezes/química , Feminino , Lactação/fisiologia , Ocratoxinas/toxicidade , Rúmen/metabolismo , Triticum/microbiologia
5.
J Physiol Pharmacol ; 56(3): 369-80, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16204760

RESUMO

In order to challenge in vivo muscle Ca2+ homeostasis and analyze consequences on mitochondrial H2O2 release (MHR) and sarcopenia, we injected Ca2+ ionophore A23187 (200 microg/kg, ip) in adult and old rats and measured gastrocnemius mass and mitochondrial Ca2+ content (MCC) using radioactive Ca2+ 48 h after injection. In a second experiment performed in old rats, we measured isocitrate dehydrogenase (ICDH) activity as an index of MCC, MHR, mitochondrial respiration, citrate synthase, COX and antioxydant enzyme activities 24 h after a 150 microg/kg injection. In adult rats, muscle mass and MCC were unchanged by A23187. In old rats, MCC increased 24 h after injection as reflected by a significant increase in ICDH activity; measured MCC tended to increase at 48 h. MHR and Mn-SOD activity were significantly increased at 24 h, and GPX activity was reduced. Muscle mass was unchanged but was negatively correlated with MCC in control and treated old rats. In conclusion, in old rats, A23187 probably induced a mitochondrial Ca2+ overload responsible for the observed increase in MHR without leading to muscle atrophy on a short term basis.


Assuntos
Envelhecimento/fisiologia , Cálcio/toxicidade , Músculo Esquelético/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Animais , Calcimicina/farmacologia , Peróxido de Hidrogênio/metabolismo , Ionóforos/farmacologia , Masculino , Mitocôndrias Musculares/efeitos dos fármacos , Mitocôndrias Musculares/metabolismo , Músculo Esquelético/efeitos dos fármacos , Tamanho do Órgão/fisiologia , Consumo de Oxigênio/efeitos dos fármacos , Ratos , Ratos Wistar
6.
Am J Physiol Endocrinol Metab ; 279(6): E1391-7, 2000 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11093928

RESUMO

Methionine transsulfuration in plasma and liver, and plasma methionine and cysteine kinetics were investigated in vivo during the acute phase of sepsis in rats. Rats were infected with an intravenous injection of live Escherichia coli, and control pair-fed rats were injected with saline. Two days after injection, the rats were infused for 6 h with [(35)S]methionine and [(15)N]cysteine. Transsulfuration was measured from the transfer rate of (35)S from methionine to cysteine. Liver cystathionase activity was also measured. Infection significantly increased (P < 0.05) the contribution of transsulfuration to cysteine flux in both plasma and liver (by 80%) and the contribution of transsulfuration to plasma methionine flux (by 133%). Transsulfuration measured in plasma was significantly (P < 0.05) higher in infected rats than in pair-fed rats (0.68 and 0.25 micromol. h(-1). 100 g(-1), respectively). However, liver cystathionase specific activity was decreased by 17% by infection (P < 0.05). Infection increased methionine flux (16%, P < 0.05) less than cysteine flux (38%, P < 0.05). Therefore, the plasma cysteine flux was higher than that predicted from estimates of protein turnover based on methionine data, probably because of enhanced glutathione turnover. Taken together, these results suggest an increased cysteine requirement in infection.


Assuntos
Metionina/farmacocinética , Sepse/metabolismo , Enxofre/farmacocinética , Animais , Anorexia/metabolismo , Cistationina gama-Liase/sangue , Cisteína/sangue , Cisteína/farmacocinética , Ingestão de Alimentos , Infecções por Escherichia coli/metabolismo , Infecções por Escherichia coli/patologia , Fígado/metabolismo , Fígado/patologia , Masculino , Metionina/sangue , Isótopos de Nitrogênio , Tamanho do Órgão , Ratos , Ratos Sprague-Dawley , Sepse/patologia , Radioisótopos de Enxofre
7.
J Endocrinol ; 163(1): 15-24, 1999 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-10495402

RESUMO

We investigated the ability of pentoxifylline (PTX) to modulate protein synthesis and degradation in the presence and absence of insulin during incubation of epitrochlearis muscle, 2 or 6 days after injection of Escherichia coli. On days 2 and 6 after infection, protein synthesis was inhibited by 25%, whereas proteolysis was enhanced by 75%. Insulin (2 nM) in vitro stimulated protein synthesis in muscles from infected rats to the same extent as in controls. The ability of insulin to limit protein degradation was severely blunted 48 h after infection. On day 6 after infection, insulin inhibited proteolysis to a greater extent than on day 2. PTX suppressed the increase in plasma concentrations of tumor necrosis factor more than 600-fold after injection of bacteria, and partially prevented the inhibition of protein synthesis and stimulation of protein degradation during sepsis. Moreover, PTX administration maintained the responsiveness of protein degradation to insulin during sepsis. Thus cytokines may influence skeletal muscle protein metabolism during sepsis, both indirectly through inhibition of the effects of insulin on proteolysis, and directly on the protein synthesis and degradation machinery.


Assuntos
Infecções por Escherichia coli/metabolismo , Insulina/uso terapêutico , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , Pentoxifilina/uso terapêutico , Inibidores de Fosfodiesterase/uso terapêutico , Análise de Variância , Animais , Técnicas de Cultura , Infecções por Escherichia coli/tratamento farmacológico , Membro Anterior , Masculino , Proteínas Musculares/biossíntese , Fenilalanina/metabolismo , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/metabolismo , Tirosina/metabolismo
8.
Am J Physiol ; 275(4): E584-93, 1998 10.
Artigo em Inglês | MEDLINE | ID: mdl-9755076

RESUMO

Skeletal muscle catabolism is a characteristic metabolic response to sepsis. We investigated the ability of physiological insulin (2 nM) or insulin-like growth factor I (IGF-I, 10 nM) concentrations to modify protein metabolism during incubation of epitrochlearis 2, 6, or 15 days after injection of live Escherichia coli. On days 2 and 6 postinfection, skeletal muscle exhibited an exacerbated negative protein balance resulting from both an inhibition in protein synthesis (25%) and an enhanced proteolysis (90%) compared with controls. By day 15 postinfection, protein balance in infected rats was significantly improved compared with either day 2 or 6. At this time, protein synthesis was augmented and protein degradation was decreased in infected rats relative to day 6. Insulin or IGF-I stimulated protein synthesis in muscles from septic and control rats in vitro to the same extent at each time point examined. The ability of insulin or IGF-I to limit protein degradation was severely blunted 48 h after infection. On day 6 postinfection, the effect of insulin or IGF-I to inhibit proteolysis was more pronounced than on day 2. Incubation with IGF-I limited proteolysis to a greater extent than insulin on both days in infected but not control rats. By day 15, insulin diminished proteolysis to the same extent as in controls. The results suggest that injection of bacteria causes fundamental derangements in protein metabolism that persist for days after infection.


Assuntos
Bacteriemia/metabolismo , Infecções por Escherichia coli/metabolismo , Fator de Crescimento Insulin-Like I/farmacologia , Insulina/farmacologia , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , Animais , Relação Dose-Resposta a Droga , Cinética , Masculino , Proteínas Musculares/biossíntese , Músculo Esquelético/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Valores de Referência
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