Consumption of Boiled, but Not Grilled, Roasted, or Barbecued Beef Modifies the Urinary Metabolite Profiles in Rats.

SCOPE: The consumption of processed meat is associated with increased risk of chronic diseases, but determining how the exposure to specific cooking processes alters the metabolome is an analytical challenge. This study aims to evaluate the impact of four typical cooking methods for beef (boiling, barbecuing, grilling, and roasting) on the urinary metabolite profiles in rats, using a non-targeted approach. METHODS AND RESULTS: Male Wistar rats (n  =  48) are fed for 3 weeks with experimental diets containing either raw or cooked (boiled, barbecued, grilled, and roasted) beef. A control group is fed with milk proteins. The 24 h-urines are analyzed using LC-MS. The consumption of boiled meat leads to the specific excretion of di- and tri-peptides (aspartyl-leucine, glycyl-aspartate, and aspartyl-prolyl-threonine) and a cyclo-prolyl-proline (p < 0.001). No singular metabolite specifically associated with the groups "grilled," "roasted," and "barbecued" meat is observed. CONCLUSION: Urinary metabolite profiles of rats fed boiled beef are clearly distinct from those of rats fed with raw, grilled, roasted, or barbecued beef. The specific metabolites include the products of non-digested proteins and may be useful as potential intake biomarkers of this meat cooking method.

Urinary Medium-Chained Acyl-Carnitines Sign High Caloric Intake whereas Short-Chained Acyl-Carnitines Sign High -Protein Diet within a High-Fat, Hypercaloric Diet in a Randomized Crossover Design Dietary Trial.

The western dietary pattern is known for its frequent meals rich in saturated fat and protein, resulting in a postprandial state for a large part of the day. Therefore, our aim was to investigate the postprandial glucose and lipid metabolism in response to high (HP) or normal (NP) protein, high-fat hypercaloric diet and to identify early biomarkers of protein intake and hepatic lipid accumulation. In a crossover design, 17 healthy subjects were randomly assigned to consume a HP or NP hypercaloric diet for two weeks. In parallel, a control group (CD; n = 10) consumed a weight-maintaining control diet. Biomarkers of postprandial lipid and glucose metabolism were measured in 24 h urine and in plasma before and following a meal challenge. The metabolic profile of urine but not plasma, showed increased excretion of 13C, carnitine and short chain acyl-carnitines after adaptation to the HP diet. Urinary excretion of decatrienoylcarnitine and octenoylcarnitine increased after adaptation to the NP diet. Our results suggest that the higher excretion of short-chain urinary acyl-carnitines could facilitate the elimination of excess fat of the HP diet and thereby reduce hepatic fat accumulation previously reported, whereas the higher excretion medium-chains acyl-carnitine could be early biomarkers of hepatic lipid accumulation.

Tissue-specific effect of colitis on protein synthesis in mice: impact of the dietary protein content.

PURPOSE: Inflammatory bowel diseases are associated with an increase in the whole-body protein turnover, thus possibly requiring an additional supply of dietary proteins. Our aim was to evaluate whether increasing dietary protein content could alleviate protein metabolism alterations in the injured splanchnic and peripheral tissues during colitis and spontaneous mucosal healing. METHODS: Mice with acute chemically induced colitis received either a normal protein (P14, 14% as energy), a moderately (P30, 30%) and a very high-protein (P53, 55%) diets. At different times after the challenge, protein synthesis rate was determined in tissues using a flooding dose of 13C valine. RESULTS: Colon, liver and spleen protein synthesis rates were significantly increased after colitis induction, while being decreased in the caecum, kidneys and muscle. Contrastingly to the two other diets, P30 diet consumption allowed faster recovery of the animals, and this coincided with a rapid resaturation of the initial protein synthesis in the colon. In the other tissues studied, the high-protein diets show different effects depending on the dietary protein content consumed and on the examined tissues, with a general trend of P53 in lowering anabolism rates. CONCLUSION: This study highlights the severe impact of acute colonic inflammation on protein metabolism in different organs. In addition, dietary protein content modulated the recovery of the initial protein synthesis rate in the various tissues following colitis induction. P30 diet consumption notably showed a better ability to alleviate protein metabolism perturbations induced by colitis, that may explain its documented beneficial effect on colon mucosal healing.

Blood 15N:13C Enrichment Ratios Are Proportional to the Ingested Quantity of Protein with the Dual-Tracer Approach for Determining Amino Acid Bioavailability in Humans.

BACKGROUND: Assessment of amino acid bioavailability is of key importance for the evaluation of protein quality; however, measuring ileal digestibility of dietary proteins in humans is challenging. Therefore, a less-invasive dual stable isotope tracer approach was developed. OBJECTIVE: We aimed to test the assumption that the 15N:13C enrichment ratio in the blood increases proportionally to the quantity ingested by applying different quantities of 15N test protein. METHODS: In a crossover design, 10 healthy adults were given a semi-liquid mixed meal containing 25 g (low protein) or 50 g (high protein) of 15N-labeled milk protein concentrate simultaneous with 0.4 g of highly 13C-enriched spirulina. The meal was distributed over multiple small portions, frequently provided every 20 min during a period of 160 min. For several amino acids, the blood 15N- related to 13C-isotopic enrichment ratio was determined at t = 0, 30, 60, 90, 120, 180, 240, 300, and 360 min and differences between the 2 meals were compared using paired analyses. RESULTS: No differences in 13C AUC for each of the measured amino acids in serum was observed when ingesting a low- or high-protein meal, whereas 15N AUC of amino acids was ∼2 times larger on the high-protein meal (P < 0.001). Doubling the intake of 15N-labeled amino acids increased the 15N:13C ratio by a factor of 2.04 ± 0.445 for lysine and a factor between 1.8 and 2.2 for other analyzed amino acids, with only phenylalanine (2.26), methionine (2.48), and tryptophan (3.02) outside this range. CONCLUSIONS: The amino acid 15N:13C enrichment ratio in the peripheral circulation increased proportionally to the quantity of 15N-labeled milk protein ingested, especially for lysine, in healthy adults. However, when using 15N-labeled protein, correction for, e.g., α-carbon 15N atom transamination is advised for determination of bioavailability of individual amino acids. This trial was registered at www.clinicaltrials.gov as NCT02966704.

Urinary Metabolomics Profiles Associated to Bovine Meat Ingestion in Humans.

SCOPE: The impact of meat consumption on human health is widely examined in nutritional epidemiological studies, especially due to the connection between the consumption of red and processed meat and the risk of colon cancer. Food questionnaires do not assess the exposure to different methods of meat cooking. This study aimed to identify biomarkers of the acute ingestion of bovine meat cooked with two different processes. METHODS AND RESULTS: Non-targeted UPLC-MS metabolite profiling was done on urine samples obtained from 24 healthy volunteers before and 8 h after the ingestion of a single meal composed of intrinsically 15 N labelled bovine meat, either cooked at 55 °C for 5 min or at 90 °C for 30 min. A discriminant analysis extension of independent components analysis was applied to the mass spectral data. After meat ingestion, the urinary excretion of 1-methylhistidine, phenylacetylglutamine, and short- and medium-chained acylcarnitines was observed. 15 N labelling was detected in these metabolites, thus confirming their origin from ingested meat. However, no difference was observed in urinary metabolomic profiles according to the meat cooking process used. CONCLUSION: Meat ingestion led to the excretion of several nitrogen-containing compounds, but although a metabolic signature was detected for meat ingestion, the impact of the cooking process was not detectable at the level of urinary metabolic signature in our experimental conditions.

Time course of fractional gluconeogenesis after meat ingestion in healthy adults: a D2O study.

In the postprandial state, glucose homeostasis is challenged by macronutrient intake, including proteins that trigger insulin secretion and provide glucose precursors. However, little is known about the postprandial response of gluconeogenesis to a protein meal. We aimed to quantify the evolution of fractional gluconeogenesis after a meat meal. Thirteen healthy subjects received oral doses of D2O. After fasting overnight, they ingested a steak (120 g). Glycemia, insulinemia, and 2H enrichments in glucose and plasma water were measured for 8 h after the meal. Fractional gluconeogenesis was assessed using the average method. Glucose was stable for 5 h and then decreased. There was a slight increase of insulin 1 h after the meal. 2H enrichment in the carbon 5 position of glucose (C5) increased after 2 h, whereas it decreased in plasma water. Consequently, fractional gluconeogenesis increased from 68.2 ± 7.2% before the meal to 75.5 ± 5.8% 8 h after the meal, the latter corresponding to 22 h without a glucose supply. These values are consistent with the exhaustion of glycogen stores after 24 h but represent the highest among values in the literature. The impact of methodological conditions is discussed.

Biosynthesis of benzylisoquinoline alkaloids in Corydalis bracteata: compartmentation and seasonal dynamics.

Numerous species of the genus Corydalis (Papaveraceae) produce a large spectrum of benzylisoquinoline alkaloids (BIA), some of which are of potential therapeutic value, but no information on sites of their biosynthesis and compartmentation is available. This study focuses on the biosynthesis, compartmentation and seasonal dynamics of BIA in Corydalis bracteata (Steph. ex Willd) Pers., a geophyte with a very short spring vegetation period, which for the rest of the year is represented by underground tubers with buds. It was found that all organs of C. bracteata contained high levels of BIA, the highest concentrations being detected in underground tuber buds in early autumn. Neither xylem nor phloem sap contained alkaloids throughout the year but BIA were present in the apoplastic wash fluid of the tuber. The absence of long-distance transport of alkaloids was confirmed by the experiment using an isotopically labeled tracer, [ring-(13)C6]-tyramine: when whole plants were fed with the tracer with via the roots, the alkaloids became labeled in the roots only and not in other organs. However, when detached roots, leaves, tubers and stems were exposed to [ring-(13)C6]-tyramine, the label was incorporated into alkaloids in all organs. We conclude that no long-distance translocation of alkaloids occurs between organs of C. bracteata, while in the tuber the cell-to-cell transport of alkaloids could occur via the apoplast. In contrast to other BIA-producing species, every organ of C. bracteata was found to be capable of de novo biosynthesis of the full complement of alkaloids.

The Role of Temperature in the Growth and Flowering of Geophytes.

Among several naturally occurring environmental factors, temperature is considered to play a predominant role in controlling proper growth and flowering in geophytes. Most of them require a "warm-cold-warm" sequence to complete their annual cycle. The temperature optima for flower meristem induction and the early stages of floral organogenesis vary between nine and 25 °C, followed, in the autumn, by a several-week period of lower temperature (4-9 °C), which enables stem elongation and anthesis. The absence of low temperature treatment leads to slow shoot growth in spring and severe flowering disorders. Numerous studies have shown that the effects of the temperature surrounding the underground organs during the autumn-winter period can lead to important physiological changes in plants, but the mechanism that underlies the relationship between cold treatment and growth is still unclear. In this mini-review, we describe experimental data concerning the temperature requirements for flower initiation and development, shoot elongation, aboveground growth and anthesis in bulbous plants. The physiological processes that occur during autumn-winter periods in bulbs (water status, hormonal balance, respiration, carbohydrate mobilization) and how these changes might provoke disorders in stem elongation and flowering are examined. A model describing the relationship between the cold requirement, auxin and gibberellin interactions and the growth response is proposed.