Amino Acid Absorption Research Articles

Effects of Different Protein Sources on Amino Acid Absorption and Plasma Appearance of Tryptophan, Large Neutral Amino Acids, and Tryptophan Metabolites in Pigs

BackgroundAbsorption of tryptophan (TRP) across the gut epithelium is potentially modulated by competing large neutral amino acids (LNAAs), which could affect the appearance of TRP and its metabolites in the bloodstream. ObjectivesThis study aimed to determine, in a growing pig model of an adult human, the absorption of TRP and other LNAAs from the gastrointestinal tract, and plasma appearance of TRP, LNAAs, and TRP metabolites, in response to dietary proteins varying in TRP content. MethodsPigs were adapted for 7 d to each of 4 diets that differed in their protein source and TRP content: 1) alpha-lactalbumin (AL; 9.95 mg TRP/g diet DM), 2) whey protein (6.59 mg TRP/g), 3) casein (3.73 mg TRP/g), or 4) zein (0.14 mg TRP/g). On day 8, pigs were euthanised after a 12-h fast (baseline), or 1, 2, 3, 4, or 6 h after they received a test meal consisting of 45 g protein, or a protein-free meal (n = 6 pigs at each time in each meal group). Tryptophan and LNAA absorption from the small intestine, and appearance of TRP, LNAAs, and TRP metabolites (melatonin, serotonin, kynurenine pathway metabolites), in the portal vein and systemic circulation, were determined. ResultsAL intake resulted in sustained elevated plasma TRP concentrations after an overnight fast. The amount of TRP absorbed was dose-dependently related to protein TRP content (P = 0.028), with fastest rates for pigs fed AL (371 mg/h). Portal and systemic plasma TRP, TRP/LNAA, and the TRP metabolites were highest (P ≤ 0.05) after AL intake, and remained above baseline levels for ∼4 h postprandially. Absorption rates of TRP correlated with postprandial plasma TRP and TRP metabolites (P ≤ 0.05). ConclusionsIn adult humans, postprandial plasma TRP and TRP metabolite concentrations can likely be modulated by the TRP content of the meal.

Analytical studies on the absorption and metabolism of amino acids by Saccharomyces species

Analytical studies on the absorption and metabolism of amino acids by <i>Saccharomyces</i> species

Comparison of gut transcriptome and bacterial composition of the yellow peach moth, Conogethes punctiferalis larvae associated with host plants adaptation

The yellow peach moth, Conogethes punctiferalis, is one of the most destructive polyphagous pests to corn crops in the Huang-Huai-Hai summer corn region of China. However, little is known about the host plant adaptation mechanism of C. punctiferalis. In this study, we analyzed the performance of C. punctiferalis on two of its favorable host plants (corn and sorghum). Then, we used RNA-seq and 16S rRNA sequencing to explore the potential adaptation mechanism of C. punctiferalis on these two host plants. Results showed that C. punctiferalis gained more fresh weight on sorghum while its total lipid and triglyceride content was significantly lower than on corn. In total, 2155 genes weredifferentially expressed (DEGs) between corn and sorghum reared C. punctiferalis. Most of the DEGs were involved in nutritional biosynthesis and metabolism including amino acid biosynthesis, protein digestion and absorption, and glycolysis. Enzymatic analyses revealed that C. punctiferalis reared on corn had higher trypsin activity but lower P450 activity than that reared on sorghum. Meanwhile, C. punctiferalis reared on corn harbored more gut bacteria, while its diversity is lower than that reared on sorghum. The potential functional prediction of the gut bacteria revealed that nutritional metabolism functions were differently enriched between two host plants of C. punctiferalis. Taken together, these findings clarify the impact of host plants on the gene expression and gut bacteria in C. punctiferalis. They also suggest that the plasticity of gene expression and gut bacteria cooperatively contribute to insect host adaptation of insects.

Whole lung lavage and GM-CSF use for pulmonary alveolar proteinosis in an infant with lysinuric protein intolerance: a case report

BackgroundLysinuric protein intolerance (LPI) is a multi-organ metabolic disorder characterized by the imbalance in absorption and excretion of cationic amino acids like lysine, ornithine and arginine. Infants with LPI typically present with recurrent vomiting, poor growth, interstitial lung disease or renal impairment. The early onset of pulmonary alveolar proteinosis (PAP) has been reported to be associated with a severe form of LPI. Treatment of PAP most commonly consists of whole-lung lavage (WLL) and in autoimmune PAP, granulocyte-macrophage colony stimulating factor (GM-CSF) administration. Nevertheless, GM-CSF therapy in LPI-associated PAP has not been scientifically justified.Case presentationWe describe the case of an 8-month-old infant presenting with respiratory failure due to LPI associated with PAP, who was twice treated with WLL; firstly, while on veno-venous ECMO assistance and then by the use of a selective bronchial blocker. After the two treatments with WLL, she was weaned from daytime respiratory support while on initially subcutaneous, then on inhaled GM-CSF therapy.ConclusionsThis case supports the notion that GM-CSF therapy might be of benefit in patients with LPI-associated PAP. Further studies are needed to clarify the exact mechanism of GM-CSF in patients with LPI-associated PAP.

Protein Ingredient Quality within Infant Formulas Impacts Plasma Amino Acid Concentrations in Neonatal Minipiglets

BackgroundInfant formulas (IFs), the only adequate substitute to human milk, are complex matrices that require numerous ingredients and processing steps that may impact protein digestion and subsequent amino acid (AA) absorption. ObjectivesThe objective was to understand the impact of the protein ingredient quality within IFs on postprandial plasma AA profiles. MethodsFour isonitrogenous and isocaloric IFs were produced at a semi-industrial scale using whey proteins from different origins (cheese compared with ideal whey) and denaturation levels (IF-A, -B, -C), and caseins with different supramolecular organizations (IF-C, -D). Ten Yucatan minipiglets (12- to 27-d-old) were used as a human infant model and received each IF for 3 d according to a Williams Latin square followed by a 2-d wash-out period. Jugular plasma was regularly sampled from 10 min preprandial to 4 h postprandial on the third day to measure free AAs, urea, insulin, and glucose concentrations. Data were statistically analyzed using a mixed linear model with diet (IFs), time, and sex as fixed factors and piglet as random factor. ResultsIFs made with cheese whey (IF-A and -B) elicited significantly higher plasma total and essential AA concentrations than IFs made with ideal whey (IF-C and -D), regardless of the pre- and postprandial times. Most of the differences observed postprandially were explained by AA homeostasis modifications. IFs based on cheese whey induced an increased plasma concentration of Thr due to both a higher Thr content in these IFs and a Thr-limiting degrading capability in piglets. The use of a nonmicellar casein ingredient led to reduced plasma content of AA catabolism markers (IF-D compared with IF-C). ConclusionsOverall, our results highlight the importance of the protein ingredient quality (composition and structure) within IFs on neonatal plasma AA profiles, which may further impact infant protein metabolism.

Systematic-Narrative Hybrid Literature Review: Crosstalk between Gastrointestinal Renin-Angiotensin and Dopaminergic Systems in the Regulation of Intestinal Permeability by Tight Junctions.

In the first part of this article, the role of intestinal epithelial tight junctions (TJs), together with gastrointestinal dopaminergic and renin-angiotensin systems, are narratively reviewed to provide sufficient background. In the second part, the current experimental data on the interplay between gastrointestinal (GI) dopaminergic and renin-angiotensin systems in the regulation of intestinal epithelial permeability are reviewed in a systematic manner using the PRISMA methodology. Experimental data confirmed the copresence of DOPA decarboxylase (DDC) and angiotensin converting enzyme 2 (ACE2) in human and rodent enterocytes. The intestinal barrier structure and integrity can be altered by angiotensin (1-7) and dopamine (DA). Both renin-angiotensin and dopaminergic systems influence intestinal Na+/K+-ATPase activity, thus maintaining electrolyte and nutritional homeostasis. The colocalization of B0AT1 and ACE2 indicates the direct role of the renin-angiotensin system in amino acid absorption. Yet, more studies are needed to thoroughly define the structural and functional interaction between TJ-associated proteins and GI renin-angiotensin and dopaminergic systems.

157 Effects of increasing dietary protein concentration on digestibility of amino acids by growing pigs

Abstract Two experiments were conducted to test the hypothesis that the standardized ileal digestibility (SID) of amino acids (AA) by growing pigs is not influenced by dietary protein concentration if all AA are supplied by soybean protein isolate (SPI) or casein. In Exp. 1, cannulated pigs [n = 12; average body weight (BW) = 39.22 kg; SD = 3.96] were allotted to a replicated 6 × 3 Youden square design with the 6 diets and 3 periods. Five diets were formulated to contain 7 to 35% SPI. An N-free diet was also used. In Exp. 2, cannulated pigs (n = 24 average BW = 23.12 kg; SD = 2.89) were allotted a quadruplicated 6 × 2 Youden square design with the 6 diets and 2 periods. Five diets were formulated to contain 6.75 to 33.75% casein. An N-free diet was also used. Ileal digesta were collected and analyzed for dry matter (DM), Cr, and AA. The statistical models included diet as a fixed variable and square, animal, and period as random effects. Polynomial contrasts were used to test linear and quadratic effects of increasing dietary protein. Results from Exp. 1 indicated that the apparent ileal digestibility (AID) of DM linearly (P = 0.001) decreased and the AID of all AA quadratically (P &amp;lt; 0.05) increased as the inclusion of SPI increased in the diets (Table 1). The SID of His and Trp was not affected by the inclusion of SPI in the diets. However, the SID of Arg, Leu, Lys, Phe, Thr, and Val linearly (P &amp;lt; 0.05) decreased, and the SID of Ile and Met tended to linearly (P &amp;lt; 0.10) decrease as the inclusion of SPI in the diets increased. Results from Exp. 2 indicated that the AID of DM tended to linearly decrease (P = 0.089) as the inclusion of casein increased in the diets (Table 2). The AID of most AA increased quadratically (P &amp;lt; 0.01) and the AID of Trp tended to increase (quadratic, P = 0.068) with increasing inclusion rate of casein. The SID of most AA was independent of casein levels. In conclusion, the anti-nutritional factors may have been increased in the diets as the inclusion of SPI increased and thus resulted in reduced SID values. However, the SID values were not influenced by increasing dietary protein if all AA were supplied by casein that does not contain anti-nutritional factors. Therefore, it is concluded that pigs have the capacity to absorb peptides and AA with the same efficiency if the protein concentration is 30% as if it is 6%. Dietary protein concentration itself, therefore, is not likely to limit AA absorption. However, even small amounts of anti-nutritional factors appear to negatively impact the SID of AA.

Alcoholic Extracts of Eleusine indica as Alternative Diuretic Regimens: A Computational Based Investigation

Diuretics are widely used in current clinical practice to increase urine production and excrete electrolytes, particularly sodium and chloride ions, without affecting the absorption of protein, vitamins, carbohydrates, or amino acids. From the time of mercury chloride and organomercurials in ancient times to now (with sulphonamides, thiazides, and furosemide), a lot has changed in the field of diuretics. However, long-term use of such synthetic diuretic agents in clinical practice produces several adverse effects, such as blurred vision, loss of appetite, stomach upset, carcinomas, headaches, phototoxic impact, weakness., etc., as has been observed from recent investigations. Natural regimens can serve as potential alternatives to using nontoxic diuretic agents. Based on long-term ethnomedicinal and biological activity records, we have explored the diuretic effects of the widely known perennial herb in Pacific Islands regions and a weed in agricultural fields, Eleusine indica (L) Gaertn phytoconstituents, on a computation platform. Therefore, we conducted a bio-assay-guided crude extraction using ethanol, followed by further gas chromatography-mass spectrometry (GC-MS) analyses of the extracted crude extracts. Further selected nine constituents (EI_1 to EI_9) carried out the diuretic potency against three putative target enzymes (ACE, KCNJ1, and SLC12A1) along with three standard drugs (VU590, TSM, and FSM) through molecular docking studies using AutoDock 4.2 software. We also predict physicochemical profiles, or Lipinski Rule of Five profiles, toxicity, and pharmacokinetics using various bioinformatics and cheminformatics tools. Based on the overall investigation, it was revealed that EI_6 [Z, Z-6,28-Heptatriacontadien-2-one] was the most potential, nontoxic, and drug-able candidate. In summary, advanced computational tools play a crucial role in selecting potential preclinical candidates within limited resources to accelerate the current drug discovery process.

Dairy Matrix Effects: Physicochemical Properties Underlying a Multifaceted Paradigm.

When food products are often considered only as a source of individual nutrients or a collection of nutrients, this overlooks the importance of interactions between nutrients, but also interactions between nutrients and other constituents of food, i.e., the product matrix. This product matrix, which can be defined as 'The components of the product, their interactions, their structural organization within the product and the resultant physicochemical properties of the product', plays a critical role in determining important product properties, such as product stability, sensory properties and nutritional and health outcomes. Such matrix effects can be defined as 'the functional outcome of specific component(s) as part of a specific product matrix'. In this article, dairy matrix effects are reviewed, with particular emphasis on the nutrition and health impact of dairy products. Such matrix effects are critical in explaining many effects of milk and dairy products on human nutrition and health that cannot be explained solely based on nutrient composition. Examples hereof include the low glycemic responses of milk and dairy products, the positive impact on dental health, the controlled amino acid absorption and the absence of CVD risk despite the presence of saturated fatty acids. Particularly, the changes occurring in the stomach, including, e.g., coagulation of casein micelles and creaming of aggregated fat globules, play a critical role in determining the kinetics of nutrient release and absorption.

How α -lactalbumin and β –casein level in infant formula influence the protein and minerals absorption properties by using Caco-2 cell model

This study investigated the differences in absorption of infant formula with different levels of α-lactalbumin (α-La) and β-casein (β-CN), and how α-La and β-CN (α+β) affect mineral absorption using the Caco-2 cell model. Compared to the control group, the free amino acid content transported to the basal side significantly increased in the fortification of α-La and β-CN, particularly in the α+β (high) group, which reached 11.343 μmol/L. Furthermore, fortification of α-La and β-CN upregulated the expression of the amino acid transporter SLC6A14 and the oligopeptide transporter PEPT1, in particular the expression of SLC6A14 in the α+β (high) group was 103% of that of the control group. In addition, fortification of α-La and β-CN enhanced the absorption of Ca, Mg, and Zn. The absorption rates of Ca, Mg, and Zn in α+β (high) group increased to 31.27%, 13.66%, and 58.59%, respectively, of which α-La contributed the most on Ca absorption. In conclusion, α-La and β-CN fortification not only increased the absorption of amino acids and peptides but also promoted the absorption of minerals, which provide evidence for the benefits of adjustment of α-La and β-CN in the infant formula.

Metabolomics and gut metagenomics profile of the healthy adults on consumption of whey protein supplemented with enzymes-probiotics blend

In this randomized, crossover, pilot clinical study, we aimed to evaluate the effect of supplementation of enzymes-probiotics blend with whey protein on the amino acid absorption and gut microbiota. Healthy subjects were supplemented with the whey protein and test i.e. Pepzyme Pro (enzymes-probiotics blend) or placebo i.e. maltodextrin for 15 days with the washout period of 30 days. Blood samples were analyzed for plasma free amino acids, insulin, and CRP. Additionally, urine nitrogen, fecal nitrogen, and gut microbiota were evaluated. On day 15, the test arm showed upward trend in rate of amino acid absorption than placebo arm within 30 min of post ingestion of protein. Moreover, rate of absorption of few essential and branched chain amino acids were significantly higher (methionine (p = 0.049), leucine (p = 0.014), isoleucine (p = 0.053)) in the test arm on day 15. Total branched chain amino acids absorption were found to be significantly higher (p ≤ 0.05) in the test arm than the placebo arm within 30 min of post ingestion on day 15. Uptrend in total amino acid absorption and Cmax, and downtrend in Tmax was observed on day 15 in the test arm. The CRP, fecal nitrogen, and urine nitrogen remained unaltered after supplementation. Microbiota profiling showed significant change in abundance of species of genus Bacteroides and phylum Bacteroidetes. Overall, metagenomics and metabolomics based assessments demonstrated that the consumption of Pepzyme Pro with whey protein could potentially improve protein digestion, amino acid absorption, and modulate gut microbiota.Clinical trial registration The clinical trial registry of India CTRI/2021/09/036169 [Registered on: 02/09/2021]

Effects of the kinetic pattern of dietary glucose release on nitrogen utilization, the portal amino acid profile, and nutrient transporter expression in intestinal enterocytes in piglets

BackgroundPromoting the synchronization of glucose and amino acid release in the digestive tract of pigs could effectively improve dietary nitrogen utilization. The rational allocation of dietary starch sources and the exploration of appropriate dietary glucose release kinetics may promote the dynamic balance of dietary glucose and amino acid supplies. However, research on the effects of diets with different glucose release kinetic profiles on amino acid absorption and portal amino acid appearance in piglets is limited. This study aimed to investigate the effects of the kinetic pattern of dietary glucose release on nitrogen utilization, the portal amino acid profile, and nutrient transporter expression in intestinal enterocytes in piglets.MethodsSixty-four barrows (15.00 ± 1.12 kg) were randomly allotted to 4 groups and fed diets formulated with starch from corn, corn/barley, corn/sorghum, or corn/cassava combinations (diets were coded A, B, C, or D respectively). Protein retention, the concentrations of portal amino acid and glucose, and the relative expression of amino acid and glucose transporter mRNAs were investigated. In vitro digestion was used to compare the dietary glucose release profiles.ResultsFour piglet diets with different glucose release kinetics were constructed by adjusting starch sources. The in vivo appearance dynamics of portal glucose were consistent with those of in vitro dietary glucose release kinetics. Total nitrogen excretion was reduced in the piglets in group B, while apparent nitrogen digestibility and nitrogen retention increased (P < 0.05). Regardless of the time (2 h or 4 h after morning feeding), the portal total free amino acids content and contents of some individual amino acids (Thr, Glu, Gly, Ala, and Ile) of the piglets in group B were significantly higher than those in groups A, C, and D (P < 0.05). Cluster analysis showed that different glucose release kinetic patterns resulted in different portal amino acid patterns in piglets, which decreased gradually with the extension of feeding time. The portal His/Phe, Pro/Glu, Leu/Val, Lys/Met, Tyr/Ile and Ala/Gly appeared higher similarity among the diet treatments. In the anterior jejunum, the glucose transporter SGLT1 was significantly positively correlated with the amino acid transporters B0AT1, EAAC1, and CAT1.ConclusionsRational allocation of starch resources could regulate dietary glucose release kinetics. In the present study, group B (corn/barley) diet exhibited a better glucose release kinetic pattern than the other groups, which could affect the portal amino acid contents and patterns by regulating the expression of amino acid transporters in the small intestine, thereby promoting nitrogen deposition in the body, and improving the utilization efficiency of dietary nitrogen.

A meta-analysis of the relationship between milk protein production and absorbed amino acids and digested energy in dairy cattle

Milk protein production is the largest draw on AA supplies for lactating dairy cattle. Prior NRC predictions of milk protein production have been absorbed protein (MP)-based and used a first-limiting nutrient concept to integrate the effects of energy and protein, which yielded poor accuracy and precision (root mean squared error [RMSE] >21%). Using a meta-data set gathered, various alternative equation forms considering MP, absorbed total EAA, absorbed individual EAA, and digested energy (DE) supplies as additive drivers of production were evaluated, and all were found to be superior in statistical performance to the first limitation approach (RMSE = 14%-15%). Inclusion of DE intake and a quadratic term for MP or absorbed EAA supplies were found to be necessary to achieve intercept estimates (nonproductive protein use) that were similar to the factorial estimates of the National Academies of Sciences, Engineering, and Medicine (2021). The partial linear slope for MP was found to be 0.409, which is consistent with the observed slope bias of -0.34 g/g when a slope of 0.67 was used for MP efficiency in a first-limiting nutrient system. Replacement of MP with the supplies of individual absorbed EAA expressed in grams per day and a common quadratic across the EAA resulted in unbiased predictions with improved statistical performance as compared with MP-based models. Based on Akaike's information criterion and biological consistency, the best equations included absorbed His, Ile, Lys, Met, Thr, the NEAA, and individual DE intakes from fatty acids, NDF, residual OM, and starch. Several also contained a term for absorbed Leu. These equations generally had RMSE of 14.3% and a concordance correlation of 0.76. Based on the common quadratic and individual linear terms, milk protein response plateaus were predicted at approximately 320 g/d of absorbed His, Ile, and Lys; 395 g/d of absorbed Thr; 550 g/d of absorbed Met; and 70 g/d of absorbed Leu. Therefore, responses to each except Leu are almost linear throughout the normal in vivo range. De-aggregation of the quadratic term and parsing to individual absorbed EAA resulted in nonbiological estimates for several EAA indicating over-parameterization. Expression of the EAA as g/100 g total absorbed EAA or as ratios of DE intake and using linear and quadratic terms for each EAA resulted in similar statistical performance, but the solutions had identifiability problems and several nonbiological parameter estimates. The use of ratios also introduced nonlinearity in the independent variables which violates linear regression assumptions. Further screening of the global model using absorbed EAA expressed as grams per day with a common quadratic using an all-models approach, and exhaustive cross-evaluation indicated the parameter estimates for BW, all 4 DE terms, His, Ile, Lys, Met, and the common quadratic term were stable, whereas estimates for Leu and Thr were known with less certainty. Use of independent and additive terms and a quadratic expression in the equation results in variable efficiencies of conversion. The additivity also provides partial substitution among the nutrients. Both of these prevent establishment of fixed nutrient requirements in support of milk protein production.

Study of amino acids absorption and gut microbiome on consumption of pea protein blended with enzymes-probiotics supplement.

The current randomized, double-blind, crossover clinical trial was conducted to evaluate changes in the amino acid absorption and gut microbiota on consumption of pea protein supplemented with an enzymes-probiotics blend (Pepzyme Pro). A total of 15 healthy subjects were instructed to take test (pea protein + Pepzyme Pro) or placebo (pea protein + maltodextrin) for 15 days with a 30-day washout period. Blood samples were analyzed for plasma-free amino acids, insulin, and C-reactive protein (CRP). Additionally, nitrogen levels in urine and feces, along with the composition of gut microbiota, were evaluated. On day 15, the test arm showed a tendency to increase the rate of absorption and total absorption (AUC) of amino acids compared with the placebo arm, though the increase was statistically insignificant. In addition, 15-day test supplementation showed a tendency to reduce Tmax of all the amino acids (statistically insignificant except alanine, p = 0.021 and glycine, p = 0.023) in comparison with the placebo supplementation. There were no changes in urine and fecal nitrogen levels as well as serum CRP levels in the test and placebo arm. The increase in serum insulin level after 4 h was statistically significant in both arms, whereas the insulin level of the placebo and test arm at 4 h was not statistically different. Supplementation showed changes with respect to Archaea and few uncharacterized species but did not show statistically significant variations in microbiome profile at the higher taxonomic levels. A study with large sample size and detailed gut microbiome analysis is warranted to confirm the results statistically as well as to characterize altered species. However, the current study could provide an inkling of a positive alteration in protein digestibility, amino acid absorption, and gut microbiome with regular consumption of protein and enzymes-probiotics blend. Clinical Trial Registration:clinicaltrials.gov/; identifier [CTRI/2021/10/037072].

Precision formulation, a new concept to improve dietary amino acid absorption based on the study of cationic amino acid transporters

Amino acid (AA) transporters (AAT) control AA cellular fluxes across membranes, contributing to maintain cellular homeostasis. In this study, we took advantage of rainbow trout metabolic feature, which highly relies on dietary AA, to explore the cellular and physiological consequences of unbalanced diets on AAT dysregulations with a particular focus on cationic AAs (CAA), frequently underrepresented in plant-based diets. Results evidenced that 24 different CAAT are expressed in various trout tissues, part of which being subjected to AA- and CAA-dependent regulations, with y+LAT2 exchanger being prone to the strongest dysregulations. Moreover, CAA were shown to control two major AA-dependent activation pathways (namely mTOR and GCN2) but at different strength according to the CAA considered. A new feed formulation strategy has been put forward to improve specifically the CAA supplemented absorption in fish together with their growth performance. Such "precision formulation" strategy reveals high potential for nutrition practices, especially in aquaculture.

Current advances for in vitro protein digestibility.

Protein is an essential macronutrient in our diet, source of nitrogen and essential amino acids, but the biological utilization of dietary protein depends on its digestibility and the absorption of amino acids and peptides in the gastrointestinal tract. The methods to define the amount and the quality of protein to meet human nutritional needs, such as the Digestible Indispensable Amino Acid Score (DIAAS), require the use of animal models or human studies. These in vivo methods are the reference in protein quality evaluation, but they are expensive and long-lasting procedures with significant ethical restrictions. Therefore, the development of rapid, reproducible and in vitro digestion methods validated with in vivo data is an old demand. This review describes the challenges of the in vitro digestion methods in the evaluation of the protein nutritional quality. In addition to the technical difficulties to simulate the complex and adaptable processes of digestion and absorption, these methods are affected by similar limitations as the in vivo procedures, i.e., analytical techniques to accurately determine bioavailable amino acids and the contribution of the endogenous nitrogen. The in vitro methods used for the evaluation of protein digestibility, with special attention on those showing comparative data, are revised, emphasizing their pros and cons. The internationally harmonized digestion protocol proposed by the INFOGEST network is being adapted to evaluate protein and amino acid digestibility. The inter-laboratory reproducibility of this protocol was demonstrated for dairy products. The in vivo/in vitro comparability results obtained to date with this protocol for several plant and animal sources are promising, but it requires an extensive validation with a wider range of foods and substrates with known in vivo digestibility. These in vitro methods will probably not be applicable to all foods, and therefore, it is important to identify their limitations, not to elude their use, but to apply them within the limits, by using the appropriate standards and references, and always as a complementary tool to in vivo tests to reduce their number.

Gut Microbiota Modulation and Amino Acid Absorption by Lactiplantibacillus plantarum TWK10 in Pea Protein Ingestion

Gut Microbiota Modulation and Amino Acid Absorption by Lactiplantibacillus plantarum TWK10 in Pea Protein Ingestion

Administration of Free Amino Acids Improves Exogenous Amino Acid Availability when Compared with Intact Protein in Critically Ill Patients: A Randomized Controlled Study

BackgroundProtein digestion and amino acid absorption appear compromised in critical illness. The provision of enteral feeds with free amino acids rather than intact protein may improve postprandial amino acid availability. ObjectiveOur objective was to quantify the uptake of diet-derived phenylalanine after the enteral administration of intact protein compared with an equivalent amount of free amino acids in critically ill patients. MethodsSixteen patients who were mechanically ventilated in intensive care unit (ICU) at risk of malabsorption received a primed continuous infusion of L-[ring-2H5]-phenylalanine and L-[ring-3,5-2H2]-tyrosine after an overnight fast. Patients were randomly allocated to receive 20 g intrinsically L-[1-13C]-phenylalanine-labeled milk protein or an equivalent amount of amino acids labeled with free L-[1-13C]-phenylalanine via a nasogastric tube over a 2-h period. Protein digestion and amino acid absorption kinetics and whole-body protein net balance were assessed throughout a 6-h period. ResultsAfter enteral nutrient infusion, both plasma phenylalanine and leucine concentrations increased (P-time < 0.001), with a more rapid and greater rise after free amino acid compared with intact protein administration (P-time × treatment = 0.003). Diet-derived phenylalanine released into the circulation was 25% greater after free amino acids compared with intact protein administration [68.7% (confidence interval {CI}: 62.3, 75.1%) compared with 43.8% (CI: 32.4, 55.2%), respectively; P < 0.001]. Whole-body protein net balance became positive after nutrient administration (P-time < 0.001) and tended to be more positive after free amino acid in provision (P-time × treatment = 0.07). ConclusionsThe administration of free amino acids as opposed to intact protein further increases postprandial plasma amino acid availability in critically ill patients, allowing more diet-derived phenylalanine to become available to peripheral tissues.This trial was registered at clinicaltrials.gov as NCT04791774.

Comparative Toxicological Effects of Perfluorooctane Sulfonate and Its Alternative 6:2 Chlorinated Polyfluorinated Ether Sulfonate on Earthworms.

High levels of 6:2 chlorinated polyfluorinated ether sulfonate (F-53B), which is a substitute for perfluorooctane sulfonate (PFOS), are detected in various environmental matrices, wildlife, and humans. Chlorinated polyfluorinated ether sulfonate has received increased attention due to its potential risk to ecosystems. However, its toxicity in the soil organisms remains unclear. In the present study, a comparative investigation was conducted on the toxicities of 6:2 Chlorinated polyfluorinated ether sulfonate (F-53B) and PFOS to the earthworm Eisenia. fetida. F-53B was significantly more acutely toxic to earthworms than PFOS, with median lethal concentrations of 1.43 and 1.83 mmol/kg dry soil (~816 and 984 mg/kg dry soil), respectively. Although both F-53B and PFOS, at 0.4 mmol/kg dry soil (=228 and 215 mg/kg dry soil) caused oxidative stress in earthworms, as evidenced by increased superoxide dismutase, peroxidase, and catalase activities as well as malondialdehyde level, the stress caused by F-53B was higher than that caused by PFOS. In transcriptomic and metabolomic studies, negative effects of PFOS and F-53B were observed on several metabolic processes in earthworms, including protein digestion and amino acid absorption, lipid metabolism, and the immune response. Compared with PFOS, F-53B exhibited a weaker disruption of lipid metabolism, comparable potency for toxicity to the immune response, and a stronger potency in extracellular matrix destruction along with apoptosis and ferroptosis induction. Hence, our data suggest that F-53B is more toxic than PFOS to earthworms. The findings provide some new insights into the potential toxicity of F-53B to soil organisms. Environ Toxicol Chem 2024;43:170-181. © 2023 SETAC.

Gastric emptying and nutrient absorption of pea protein products differing in heat treatment and texture: A randomized in vivo crossover trial and in vitro digestion study

Pea protein is an interesting alternative for animal-based proteins due to its good availability, low cost, and relatively balanced amino acid (AA) profile. Its digestibility may be affected by heat treatment and food texture. Our aim was to study invivo AA absorption kinetics and gastric behavior of pea protein products differing in heat treatment and texture and compare this with invitro digestion. We included fourteen males in this randomized crossover trial with three iso-caloric and iso-volumetric treatments: a 420-mL heated drink, 420-mL unheated drink and 105-g heated gel (semi-solid) consumed with 315 mL water, all containing 20 g pea protein. Gastric MRI scans were made until 90 min post-prandial. Blood samples were collected at baseline and up to 5 h. All treatments were tested with an invitro digestion model (INFOGEST). Heat treatment did not alter AA absorption kinetics and gastric emptying. Time to maximum peak was delayed for the gel treatment (total AAs: 66.9 versus 48.0 min for both drinks, essential AAs: 75.4 versus 50.0 and 46.6 min for the drinks). For the gel treatment initial emptying was faster due to the rapid passage of water. Invitro, the degree of hydrolysis was highest for the unheated drink in the gastric phase and for the gel treatment in the intestinal phase. In conclusion, heat treating pea protein products does not affect digestion. In contrast, texture of pea protein products can be altered to influence the rate of gastric emptying and AA absorption without affecting total AA absorption.