Standardized ileal amino acid digestibility and performance indices in pullets and laying hens fed expeller-pressed Canadian prairie soybean meal.

Soybean meals (SBM) from different locations differ in their protein content, subsequently influencing their amino acid (AA) profile. In this study, standardized ileal digestibility (SID) of AA and growth or production performance were evaluated in pullets and hens fed SBM derived from soybean grown in Western Canada, primarily Manitoba (MB) labelled as A-, B- and C-SBM compared with that from Eastern Canada (Ontario, ON-SBM) and contained 38.3 ± 0.44, 38.6 ± 0.61, 39.4 ± 0.49, or 44.0 ± 0.87% CP, respectively. A N-free diet was used to determine basal ileal endogenous losses of AA. The study included the grower, developer, and layer phases (9-12, 13-16, and 44/59-64-wk old birds, respectively). Although a lower (P = 0.029) SID for cysteine was noted in the grower phase for the C-SBM compared with other SBM, the developer phase had higher (P < 0.05) SID for methionine, phenylalanine, cysteine (more by 4.4, 2.4 and 7.2% units, respectively) on average for SBM samples from MB compared with the ON-SBM. Regardless the source of the SBM, no difference in SID of AA was noted in the layer phase. Overall, in all phases the SID values of most AA in the SBM from MB were comparable with the ON-SBM, which may be linked to higher values of these AA per unit of protein content in the former source. In addition, the growth performance including feed intake, BW gain and feed conversion ratio in pullets, and egg production/quality in layers were similar between treatments. These findings show that the MB-SBM have a comparable feeding value with the ON-SBM, hence represent a suitable alternative protein source for poultry.

Identification and Characterization of a Pepsin- and Chymotrypsin-Resistant Peptide in the α Subunit of the 11S Globulin Legumin from Common Bean (Phaseolus vulgaris L.).

The 11S globulin legumin typically accounts for approximately 3% of the total protein in common beans (Phaseolus vulgaris). It was previously reported that a legumin peptide of approximately 20 kDa is resistant to pepsin digestion. Sequence prediction suggested that the pepsin-resistant peptide is located at the C-terminal end of the α-subunit, within a glutamic acid-rich domain, overlapping with a chymotrypsin-resistant peptide. Using purified legumin, the peptide of approximately 20 kDa was found to be resistant to pepsin digestion in a pH-dependent manner, and its location was determined by two-dimensional gel electrophoresis and LC-MS-MS. The location of the chymotrypsin-resistant peptide was confirmed by immunoblotting with peptide-specific polyclonal antibodies. The presence of a consensus site for proline hydroxylation and arabinosylation, the detection of hydroxyproline residues, purification by lectin affinity chromatography, and a difference in electrophoretic migration between the chymotrypsin- and pepsin-resistant peptides suggest the presence of a large O-glycan within these peptides.

Splenic gene expression of cytokines at multiple time points following lipopolysaccharide challenge in layers.

OBJECTIVE: To investigate inflammatory responses to lipopolysaccharide (LPS) injection in layers. ANIMALS: 33 40-week-old laying hens were used. METHODS: 30 laying hens were divided into 2 groups: the first group was injected with 8 mg/kg LPS, while the second group was injected with sterile saline. At the start of the study, 3 birds served as a baseline and were used as the time 0 controls for both the saline and LPS-treated groups. Blood and spleen tissues were collected at 0 (before) and 1, 2, 3, 4, and 6 hours after injection. RESULTS: LPS administration increased splenic mRNA levels of IL-1ß, IL-2, IL-6, IL-8, IL-10, interferon-γ, and tumor necrosis factor-α (P < .001) and serum IL-6 levels (P < .01) compared to saline injection. The mRNA expression of most cytokine genes increased rapidly toward peak values within 2 hours after the LPS injection, and then the difference between the saline and LPS treatments got smaller as time went on; serum IL-6 reached its highest concentration 2 hours after LPS administration. The magnitude of LPS-induced upregulation of gene expression was the highest for IL-6, followed by IL-1ß and IL-8, and tumor necrosis factor-α was the least affected. CLINICAL RELEVANCE: The temporal and quantitative profile of these inflammatory mediators generated from this study provides valuable information in identifying the optimal time window and appropriate biomarkers for LPS-induced inflammation, which has significant implications in evaluating the effects of interventions on the immune system of chickens.

Effects of genotype, environment and their interaction on protein and amino acid contents in soybeans.

Soybean [Glycine max (L.) Merr.] is an important source of protein and oil. Genotype, environment and the interaction of genotype × environment influence the protein composition in soybean seeds. The main objectives of this study were to i) study the influence of genotype, environment and their interaction on soybean protein and amino acid contents; ii) evaluate the stability of soybean genotype across various environments, with a focus on soybeans grown in a northern latitude; and iii) identify the correlation between crude protein and critical amino acid value (CAAV). Twenty-three soybean genotypes were grown at four locations in Manitoba, Canada for two years (2018 and 2019). Soybean seeds were analyzed for protein and amino acids (nine essential amino acids and cysteine). The effects of genotype, environment and genotype × environment interaction on all traits were significant (P < 0.05). Genotype and environments explained the main part of variation for all traits. G13 and G15 cultivars performed better in favorable environments (bi > 1), and G22 cultivar showed greater resistance to environmental change. Protein and amino acids responded differently to various environments, but the optimal environments for greater soybean protein and amino acid remains to be established. The CAAV had a negative linear relationship with protein content in soybean (k = -0.17). This work expands our knowledge of the factors impacting the protein quality of soybeans grown in northern latitudes.

The Effect of Increasing Concentrations of Omega-3 Fatty Acids from either Flaxseed Oil or Preformed Docosahexaenoic Acid on Fatty Acid Composition, Plasma Oxylipin, and Immune Response of Laying Hens.

BACKGROUND: There is a lack of nutrition guidelines for the feeding of omega-3 polyunsaturated fatty acids (PUFA) to laying hens. Knowledge as to whether the type and concentrations of α-linolenic acid (ALA) and/or docosahexaenoic acid (DHA) in the diet can make a difference to the birds' immune responses when subjected to a lipopolysaccharide (LPS) challenge is limited. OBJECTIVES: The study was designed to determine the potential nutritional and health benefits to laying hens when receiving dietary omega-3 PUFA from either ALA or DHA. METHODS: A total of 80 Lohmann LSL-Classic (white egg layer, 20 wk old) were randomly assigned to 1 of 8 treatment diets (10 hens/treatment), provided 0.2%, 0.4%, 0.6%, or 0.8% of total dietary omega-3 PUFA, provided as either ALA-rich flaxseed oil or DHA-enriched algal biomass. After an 8-wk feeding period, the birds were challenged with Escherichia coli-derived LPS (8 mg/kg; i.v. injection), with terminal sample collection 4 h after challenge. Egg yolk, plasma, liver, and spleen samples were collected for subsequent analyses. RESULTS: Increasing dietary omega-3 supplementation yielded predictable responses in egg yolk, plasma, and liver fatty acid concentrations. Dietary intake of ALA contributed mainly to ALA-derived oxylipins. Meanwhile, eicosapentaenoic acid- and DHA-derived oxylipins were primarily influenced by DHA dietary intake. LPS increased the concentrations of almost all the omega-6 PUFA-, ALA-, and DHA-derived oxylipins in plasma and decreased hepatic mRNA expression of COX-2 and 5-LOX (P < 0.001) involved in the biosynthesis of oxylipins. LPS also increased mRNA expression of proinflammatory cytokine IFN-γ and receptor TLR-4 (P < 0.001) in the spleen. CONCLUSIONS: These results revealed that dietary intake of ALA and DHA had unique impacts on fatty acid deposition and their derived oxylipins and inflammatory responses under the administration of LPS in laying hens.

Quantitative trait loci associated with amino acid concentration and in vitro protein digestibility in pea (Pisum sativum L.).

With the expanding interest in plant-based proteins in the food industry, increasing emphasis is being placed on breeding for protein concentration and quality. Two protein quality traits i.e., amino acid profile and protein digestibility, were assessed in replicated, multi-location field trials from 2019 to 2021 in pea recombinant inbred line population PR-25. This RIL population was targeted specifically for the research of protein related traits and its parents, CDC Amarillo and CDC Limerick, had distinct variations in the concentration of several amino acids. Amino acid profile was determined using near infrared reflectance analysis, and protein digestibility was through an in vitro method. Several essential amino acids were selected for QTL analysis, including lysine, one of the most abundant essential amino acids in pea, and methionine, cysteine, and tryptophan, the limiting amino acids in pea. Based on phenotypic data of amino acid profiles and in vitro protein digestibility of PR-25 harvested in seven location-years, three QTLs were associated with methionine + cysteine concentration, among which, one was located on chromosome 2 (R2 = 17%, indicates this QTL explained 17% phenotypic variation of methionine + cysteine concentration within PR-25), and two were located on chromosome 5 (R2 = 11% and 16%). Four QTLs were associated with tryptophan concentration and are located on chromosome 1 (R2 = 9%), chromosome 3 (R2 = 9%), and chromosome 5 (R2 = 8% and 13%). Three QTLs were associated with lysine concentration, among which, one was located on chromosome 3 (R2 = 10%), the other two were located on chromosome 4 (R2 = 15% and 21%). Two QTLs were associated with in vitro protein digestibility, one each located on chromosomes 1 (R2 = 11%) and 2 (R2 = 10%). QTLs associated with in vitro protein digestibility, and methionine + cysteine concentration on chromosome 2 were identified to be co-localized with known QTL for total seed protein concentration in PR-25. QTLs associated with tryptophan and methionine + cysteine concentration co-localized on chromosome 5. The identification of QTLs associated with pea seed quality is an important step towards marker-assisted selection of breeding lines with improved nutritional quality, which will further boost the competitiveness of pea in plant-based protein markets.

Methionine and cysteine oxidation are regulated in a dose dependent manner by dietary Cys intake in neonatal piglets receiving enteral nutrition.

Methionine (Met) is an indispensable amino acid (AA) in piglets. Met can synthesize cysteine (Cys), and Cys has the ability to reduce the Met requirement by 40% in piglets. However, whether this sparing effect on Met is facilitated by downregulation of Cys synthesis has not been shown. This study investigated the effects of graded levels of Cys on Met and Cys oxidation, and on plasma AA concentrations. Piglets (n = 32) received a complete elemental diet via gastric catheters prior to being randomly assigned to one of the eight dietary Cys levels (0, 0.05, 0.1, 0.15, 0.2, 0.25, 0.40, 0.50 g kg-1d-1) with an adequate Met concentration (0.25g kg-1d-1). Constant infusion of L-[1-14C]-Met and L-[1-14C]-Cys were performed for 6 h on d 6 and d 8 to determine Met and Cys oxidation, respectively. Met oxidation decreased as Cys intake increased (P<0.05). At higher Cys intakes (0.15 to 0.5g kg-1d-1), Met oxidation decreased (P<0.05) at a slower rate. Cys oxidation was similar (P>0.05) among dietary Cys intakes; however, a significant polynomial relationship was observed between Cys oxidation and intake (P<0.05, R2 = 0.12). Plasma Met concentrations increased (P<0.05) linearly with increasing levels of dietary Cys, while plasma Cys concentrations changed (P<0.05) in a cubic manner and the highest concentrations occurred at the highest intake levels. Increasing dietary levels of Cys resulted in a reduction in Met oxidation until the requirement for the total sulfur AA was met, indicating the sparing capacity by Cys of Met occurs through inhibition of the transsulfuration pathway in neonatal piglets.

Alteration of the dietary methionine: Cysteine ratio modulates the inflammatory response to an inter-peritoneal injection of lipopolysaccharide in wistar rats.

Sulphur amino acids (SAA) are essential for multiple physiological/metabolic processes, with the ratio of dietary methionine: cysteine (Met:Cys) being an important contributor to pro-inflammatory responses, including TNF-α activity. The current study was designed to determine the effect an altered dietary SAA ratio, and the resulting reliance on the transsulfuration pathway to supply Cys, will have on the inflammatory response. In the present study, 100 µg/kg of an intraperitoneal (IP) injection of lipopolysaccharide (LPS) was used as a model for systemic inflammation. Male Wistar rats were randomized to one of two amino acid-defined diets, (100Met:0Cys or 50Met:50Cys) and subdivided to receive either IP LPS or saline injections. LPS significantly increased total plasma Cys, homocysteine (Hcy) and glutathione (GSH) 240 min post-IP injection in rats fed a 50Met:50Cys ratio compared to other treatments. The TNF-α area under the curve for LPS-treated rats consuming a dietary 50Met:50Cys ratio was significantly higher (P < .004) compared to those consuming a dietary 100Met:0Cys ratio. A significant increase in the percentage of leukocytes that were neutrophils was observed in rats injected with LPS when compared to saline with no effect of diet. These results demonstrate that an alteration of the dietary Met:Cys ratio did not attenuate the inflammatory response to an IP injection of LPS in Wistar rats; however, a diet with a balanced Met:Cys ratio increased concentrations of Cys and GSH which may result in a more rapid response to an LPS challenge.

Impact of alcohol washing on the flavour profiles, functionality and protein quality of air classified pea protein enriched flour.

In this study the potential of aqueous solvent washing on removing off-flavours in air classified pea protein-enriched flour (PPEF) was investigated. Unpleasant flavour compounds are one of the main deterrents to the application of pulses. PPEF was treated with ethanol or isopropanol at three different concentrations (20%, 50%, and 80%) to remove the volatiles related to unpleasant beany, earthy and astringent flavours. Headspace solid phase microextraction followed by GC-MS was used to identify the flavour compounds in untreated and treated PPEF. Besides the flavour profile, changes to their proximate composition, colour, functionality and protein quality were compared among untreated and treated samples. Higher concentrations of ethanol and isopropanol (50% and 80%) showed greater effectiveness in removing flavour compounds by reducing the total peak area by 82%-94%. Protein content in all treated samples (58.2%-64.3% d.b.) increased compared to untreated PPEF (55.5%) as a result of purification due to the decrease in ash, lipid and carbohydrate content. However, alcohol treatment reduced the protein solubility and oil holding capacity in all samples by 38.3%-75.9%, and 16.7%-30.2%, respectively. Although in vitro protein digestibility was improved with the solvent treatments, the amino acid scores of those samples became lower (i.e., reduced levels of methionine, cysteine or tryptophan) resulting in up to a 27.8% reduction in in vitro protein digestibility correct amino acid scores. Both ethanol and isopropanol at 50% and 80% concentration proved to be effective in removing flavour compounds in PPEF with some modifications on the chemical compositions, protein functionalities and quality.

Methionine restriction leads to hyperhomocysteinemia and alters hepatic H2S production capacity in Fischer-344 rats.

Dietary methionine restriction (MR) increases lifespan in several animal models. Despite low dietary intake of sulphur amino acids, rodents on MR develop hyperhomocysteinemia. On the contrary, MR has been reported to increase H2S production in mice. Enzymes involved in homocysteine metabolism also take part in H2S production and hence, in this study, the impact of MR on hyperhomocysteinemia and H2S production capacity were investigated using Fischer-344 rats assigned either a control or a MR diet for 8 weeks. The MR animals showed elevated plasma homocysteine accompanied with a reduction in liver cysteine content and methylation potential. It was further found that MR decreased cystathionine-ß-synthase (CBS) activity in the liver, however, MR increased hepatic cystathionine-γ-lyase (CGL) activity which is the second enzyme in the transsulfuration pathway and also participates in regulating H2S production. The relative contribution of CGL in H2S production increased concomitantly with the increased CGL activity. Additionally, hepatic mercaptopyruvate-sulphur-transferase (MPST) activity also increased in response to MR. Taken together, our results suggest that reduced CBS activity and S-Adenosylmethionine availability contributes to hyperhomocysteinimia in MR animals. Elevated CGL and MPST activities may provide a compensatory mechanism for maintaining hepatic H2S production capacity in response to the decreased CBS activity.

Folic Acid Supplementation Attenuates Chronic Hepatic Inflammation in High-Fat Diet Fed Mice.

Nonalcoholic fatty liver disease (NAFLD) is the most common form of chronic liver disease worldwide. Hepatic inflammation is an important pathogenic mediator of NAFLD. There is currently no pharmacological agent approved for the treatment of NAFLD. Folic acid is a water-soluble B vitamin that has been shown to have lipid-lowering and antioxidant effects. The objective of this study was to investigate the effect of folic acid supplementation on hepatic inflammation and to identify the underlying mechanisms. Male C57BL/6 J mice were fed a control diet (10% kcal fat), a high-fat diet (HFD) (60% kcal fat), or a HFD supplemented with folic acid (26 mg/kg diet) for 8 weeks. HFD feeding led to increased body mass gain, lipid accumulation, activation of transcription factor nuclear factor-κB (NF-κB), and elevation of inflammatory cytokine gene expression in the liver. Folic acid supplementation attenuated hepatic lipid accumulation and aggregation of inflammatory foci induced by HFD feeding. This was associated with a significant reduction of NF-κB activation and inflammatory cytokine expression. These results suggest that the hepatoprotective effect of folic acid in NAFLD may be attributed, in part, to its anti-inflammatory action.

Recent Developments in Folate Nutrition.

The term folate (vitamin B9) refers to a group of water-soluble compounds that are nutritionally essential for the support of optimal human health and development. Folates participate in numerous one-carbon transfer reactions, including the methylation of important biomolecules (lipids, amino acids, DNA). A deficiency of folate leads to pathological outcomes including anemia and impairments in reproductive health and fetal development. Due to the linkage of impaired folate status with an increased prevalence of neural tube defects (NTDs) in babies, several jurisdictions required the fortification of the food supply with folic acid, a synthetic and stable form of folate. Data from the postfortification era have provided strong evidence for the reduction of NTDs due to folic acid fortification. However, concern is now growing with respect to the amount of synthetic folic acid within the human food supply. Excess folic acid intake has been linked to a masking of vitamin B12 deficiency, and concerns regarding the promotion of folate-sensitive cancers, including colorectal cancer. New strategies to ensure the supply of optimal folate to at-risk populations may be needed, including the use of biofortification approaches, in order to address recent concerns.

Interactions between canola meal and flaxseed oil in the diets of White Lohmann hens on fatty acid profile and sensory characteristics of table eggs.

The current study was designed to assess the fatty acid composition and sensory attributes of eggs procured from hens consuming diets containing canola meal (CM) and/or flax oil (FO). A total of 96 group-caged White Lohmann hens received 1 of 4 isonitrogenous and isoenergetic diets for a period of 4 weeks. Diets were arranged in a 2 × 2 factorial design, containing 24% canola meal, 7.5% flax oil, both, or neither (control). All yolk fatty acids were affected by flax oil inclusion, with the exception of stearic acid (SA) and docosapentaenoic acid (DPA). Only SA was affected by CM inclusion. Additionally, significant interactions between CM and FO were observed for linoleic acid (LA) and total omega-6 polyunsaturated fatty acids (PUFA), with DPA approaching significance (P = 0.069). Trained panelists (n = 8) evaluated 7 aroma ('egg', 'creamy', 'buttery', 'salty', 'sweet', 'barny', and 'oceanic') and 6 flavor ('egg', 'creamy', 'buttery', 'salty', 'brothy', and 'oceanic') attributes of cooked egg product. No significant differences (P > 0.05) in aroma attributes were found between eggs from different dietary treatments. However, egg, creamy, buttery, and oceanic flavors were significantly different between the dietary treatments (P < 0.05). While oceanic flavor significantly increased with inclusion of FO, egg and creamy flavors showed a significant decrease (P < 0.05). Although CM addition alone did not result in significant sensory changes, the pairing of CM and FO resulted in even greater sensory changes than using FO alone, specifically with regard to egg flavor. Results from partial least squares analyses showed a strong association between oceanic flavor and omega-3 PUFA. Oppositely, egg, creamy, and buttery flavors were more correlated with the presence of omega-6 PUFA and palmitic acid. This experiment provides evidence that the interaction between CM and FO in the White Lohmann hen diet results in sensory changes of cooked eggs associated in part with changes in yolk fatty acid content.

Investigation of vitamin B6 inadequacy, induced by exposure to the anti-B6 factor 1-amino D-proline, on plasma lipophilic metabolites of rats: a metabolomics approach.

PURPOSE: Vitamin B6 status in the body is affected by several factors including dietary supply of the antivitamin B6 factor, 1-amino D-proline (1ADP), which is present in flaxseed. Owing to the prevalence of moderate B6 deficiency in the general population, a co-occurrence of 1ADP may lead to a further deterioration of B6 status. To this end, we applied a nontargeted metabolomics approach to identify potential plasma lipophilic biomarkers of deleterious effect of 1ADP on moderately vitamin B6-deficient rats using a high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. METHODS: Twenty-four rats were fed with a semi-purified diet containing pyridoxine·HCl (PN·HCl) either 7 mg/kg diet (optimal B6) or 0.7 mg/kg diet (moderate B6). The rats were divided into four treatments (n = 6), and one treatment in each B6 diet group was also fed ad libitum with 10 mg/kg diet of synthetic 1ADP. After 5 weeks of study, plasma was collected from the rats and lipophilic metabolites were extracted using acetonitrile as a solvent for analysis. RESULTS: Ten potential plasma lipophilic biomarkers were identified out of >2500 detected entities, which showed significant differences between the treatments. Plasma glycocholic acid, glycoursodeoxycholic acid, murocholic acid, N-docosahexaenoyl GABA, N-arachidonoyl GABA, lumula, nandrolone and orthothymotinic acid concentrations were significantly elevated, while plasma cystamine and 3-methyleneoxindole concentrations were significantly reduced as a result of either low B6 status or 1ADP or their interaction. CONCLUSION: Changes in these metabolites revealed a potential defect in pathways linked with the biosynthesis and metabolism of bile acid components, N-acyl amino acids, analgesic androgens, anti-inflammatory and neuroprotective molecules. We also noted that the changes in these biomarkers can be alleviated by the application of adequate vitamin B6.

Tyrosol Attenuates High Fat Diet-Induced Hepatic Oxidative Stress: Potential Involvement of Cystathionine ß-Synthase and Cystathionine γ-Lyase.

The Mediterranean diet is known for its cardioprotective effects. Recently, its protective qualities have also been reported in patients with non-alcoholic fatty liver disease (NAFLD). Oxidative stress is one of the important factors responsible for the development and progression of NAFLD. Hydrogen sulfide (H2S), a multifaceted gasotransmitter, has emerged as a potential therapeutic target in NAFLD. Cystathionine ß-synthase (CBS) and cystathionine γ-lyase (CSE) are major enzymes responsible for endogenous H2S synthesis. Since oxidative stress contributes to NAFLD pathogenesis, the objective of this study was to investigate the effect of tyrosol, a major compound in olive oil and white wine, on high fat diet-induced hepatic oxidative stress and the mechanisms involved. Mice (C57BL/6) were fed for 5 weeks with a control diet (10 % kcal fat), a high fat diet (60 % kcal fat, HFD) or a HFD supplemented with tyrosol. High fat diet feeding induced hepatic oxidative stress, as indicated by the significant increase in lipid peroxidation and NADPH oxidase activity. Tyrosol supplementation significantly increased hepatic CBS and CSE expression and H2S synthesis in high fat diet-fed mice. Such effects were associated with the attenuation of high fat diet-induced hepatic lipid peroxidation and the restoration of the redox equilibrium of the antioxidant glutathione. Tyrosol also inhibited palmitic acid-induced oxidative stress in hepatocytes (HepG2 cells). These results suggest that the antioxidant properties of tyrosol may be mediated through functional changes in CBS and CSE activity, which might contribute to the hepatoprotective effect of the Mediterranean diet.

A Vitamin B-6 Antagonist from Flaxseed Perturbs Amino Acid Metabolism in Moderately Vitamin B-6-Deficient Male Rats.

BACKGROUND: Pyridoxal 5'-phosphate (PLP) plays a crucial role as a cofactor in amino acid metabolism. There is a prevalence of moderate vitamin B-6 deficiency in the population that may be exacerbated through the ingestion of 1-amino d-proline (1ADP), a vitamin B-6 antagonist found in flaxseed. OBJECTIVE: Given prior evidence of the impact of synthetic 1ADP on indexes of pyridoxine metabolism, the current study was designed to investigate the effects of 1ADP derived from flaxseed on amino acid metabolism in moderately vitamin B-6-deficient rats. METHODS: Male weanling rats (n = 8/treatment) consumed a semipurified diet containing either 7 mg pyridoxine hydrochloride/kg diet [optimum vitamin B-6 (OB)] or 0.7 mg pyridoxine hydrochloride/kg diet [moderately vitamin B-6 deficient (MB)], each with 0 or 10 mg vitamin B-6 antagonist/kg diet, in either a synthetic form (1ADP) or as a flaxseed extract (FE), for 5 wk. At the end of the experiment, plasma vitamin B-6 and amino acid concentrations and the activities of hepatic PLP-dependent enzymes were analyzed. RESULTS: Compared with the MB control group, plasma PLP concentrations were 26% and 69% lower, respectively, in the MB+FE and MB+1ADP rats (P ≤ 0.001). In the MB+FE group, the plasma cystathionine concentration was 100% greater and the plasma α-aminobutyric acid and glutamic acid concentrations were 59% and 30% lower, respectively, than in the MB control group. Both synthetic 1ADP and FE significantly (P < 0.001) inhibited the in vitro hepatic activities of 2 PLP-dependent enzymes, cystathionine ß-synthase (up to 44%) and cystathionine γ-lyase (up to 60%), irrespective of vitamin B-6 concentrations. Because of vitamin B-6 antagonist exposure, observed perturbations in plasma biomarkers and hepatic enzyme activities were not evident or of lesser magnitude in rats consuming adequate vitamin B-6. CONCLUSION: The current data from a rat model provide evidence that a vitamin B-6 antagonist now prevalent in the human food supply may pose challenges to individuals of moderate vitamin B-6 status.

Folic acid supplementation during high-fat diet feeding restores AMPK activation via an AMP-LKB1-dependent mechanism.

AMPK is an endogenous energy sensor that regulates lipid and carbohydrate metabolism. Nonalcoholic fatty liver disease (NAFLD) is regarded as a hepatic manifestation of metabolic syndrome with impaired lipid and glucose metabolism and increased oxidative stress. Our recent study showed that folic acid supplementation attenuated hepatic oxidative stress and lipid accumulation in high-fat diet-fed mice. The aim of the present study was to investigate the effect of folic acid on hepatic AMPK during high-fat diet feeding and the mechanisms involved. Male C57BL/6J mice were fed a control diet (10% kcal fat), a high-fat diet (60% kcal fat), or a high-fat diet supplemented with folic acid (26 mg/kg diet) for 5 wk. Mice fed a high-fat diet exhibited hyperglycemia, hepatic cholesterol accumulation, and reduced hepatic AMPK phosphorylation. Folic acid supplementation restored AMPK phosphorylation (activation) and reduced blood glucose and hepatic cholesterol levels. Activation of AMPK by folic acid was mediated through an elevation of its allosteric activator AMP and activation of its upstream kinase, namely, liver kinase B1 (LKB1) in the liver. Consistent with in vivo findings, 5-methyltetrahydrofolate (bioactive form of folate) restored phosphorylation (activation) of both AMPK and LKB1 in palmitic acid-treated HepG2 cells. Activation of AMPK by folic acid might be responsible for AMPK-dependent phosphorylation of HMG-CoA reductase, leading to reduced hepatic cholesterol synthesis during high-fat diet feeding. These results suggest that folic acid supplementation may improve cholesterol and glucose metabolism by restoration of AMPK activation in the liver.

Oral exposure to the anti-pyridoxine compound 1-amino D-proline further perturbs homocysteine metabolism through the transsulfuration pathway in moderately vitamin B6 deficient rats.

Pyridoxal 5'-phosphate (PLP; a B6 vitamer) serves as an important cofactor in a myriad of metabolic reactions, including the transsulfuration (TS) pathway, which converts homocysteine (Hcy) to cysteine. While overt vitamin B6 deficiency is rare, moderate deficiency is common and may be exacerbated by anti-pyridoxine factors in the food supply. To this end, we developed a model of moderate B6 deficiency and a study was conducted to examine the in vivo effect of 1-amino D-proline (1ADP), an anti-pyridoxine factor found in flaxseed, on indices of Hcy metabolism through the TS pathway in moderately B6 deficient rats. Male weaning rats received a semi-purified diet containing either 7 mg/kg (control; CD) or 0.7 mg/kg (moderately deficient; MD) diet of pyridoxine·hydrochloride (PN∙HCl), each with 1 of 4 levels of 1ADP, viz. 0, 0.1, 1 and 10 mg/kg diet for 5 weeks. Perturbations in vitamin B6 biomarkers were more pronounced in the MD group. Plasma PLP was significantly reduced, while plasma Hcy (8-fold) and cystathionine (11-fold) were increased in rats consuming the highest amount of 1ADP in the MD group. The activities of hepatic cystathionine ß-synthase and cystathionine γ-lyase enzymes were significantly reduced in rats consuming the highest 1ADP compared to the lowest, for both levels of PN∙HCl. Dilation of hepatic central veins and sinusoids, mild steatosis and increased liver triglycerides were present in MD rats consuming the highest 1ADP level. The current data provide evidence that the consumption of an anti-pyridoxine factor linked to flaxseed may pose a risk for subjects who are moderate/severe vitamin B6 deficient.

Fatty acid profile and sensory characteristics of table eggs from laying hens fed hempseed and hempseed oil.

UNLABELLED: Hempseed (HS) is rich in omega-3 polyunsaturated fatty acids, with approximately 17% of total fatty acids as alpha-linolenic acid. As such, HS and its oil may be used in hen diet formulations to produce eggs enriched in essential fatty acids. Because omega-3 eggs have the potential for unpleasant aromas and flavors, the current study was designed to assess the fatty acid profile and sensory attributes of eggs procured from hens consuming diets containing hempseed oil (HO) or HS. A total of 48 individually caged White Bovan hens received 1 of 6 diets containing 4%, 8%, 12% HO, 10%, 20% HS or 0% hemp (w/w) for 12 wk. Total omega-3 polyunsaturated fatty acid content was highest in the 12% HO group (15.3 mg/g of yolk) compared to the control (2.4 mg/g of yolk). Trained panellists (n= 8) found no significant differences (P≥ 0.05) in aroma or flavor between cooked eggs from different dietary treatments, with the exception of sweet flavor. The 4% HO group yielded the least sweet eggs compared to the 20% HS group, which was highest. For yolk color, L*, a*, and b* values (Mean ± SEM) for control eggs were 61.2 ± 0.10, 1.1 ± 0.05, and 43.0 ± 0.22, respectively. Addition of hemp led to significant (P < 0.001) reductions in L*, and significant increases in a* and b*, with the largest changes observed in the 20% HS treatment (L*= 58.7 ± 0.10; a*= 5.8 ± 0.05; b*= 60.5 ± 0.22). The results show that hemp use in hen diets leads to increased omega-3 polyunsaturated fatty acid content and color intensity of egg yolks, but does not have adverse effects on the sensory profiles of the cooked eggs. PRACTICAL APPLICATION: This study provides evidence that HS and hempseed oil (HO) can safely be utilized as feed ingredients for laying hens to produce table eggs that are enriched in essential fatty acids. Additionally, the eggs procured from these hens had similar aroma and flavor compared to eggs from hens not fed any hemp. The greater the dietary hemp inclusion, the more pigmented the resulting yolks became in terms of darkness, redness, and yellowness.

Glutathione and riboflavin status in supplemented patients undergoing home nocturnal hemodialysis versus standard hemodialysis.

BACKGROUND: Patients on conventional hemodialysis (HD) have elevated markers of oxidative stress and chronic inflammation, which may contribute to a high prevalence of cardiovascular disease. Glutathione (GSH), an important intracellular antioxidant, requires cysteine as a rate-limiting amino acid for its synthesis and riboflavin for its regeneration. OBJECTIVES: We aimed to examine whether erythrocyte GSH (eGSH) concentrations and riboflavin status are influenced by the increased dialysis dose provided to vitamin-supplemented patients receiving home nocturnal hemodialysis (HNHD) (6-8 hours/session, 5-7 nights/week) compared with patients on standard hemodialysis (SHD) (4 hours/session, 3 days/week). METHOD: This was a cross-sectional comparative study involving 30 patients undergoing SHD or HNHD regimens and a group of 15 healthy control subjects (HC). We measured eGSH concentration by liquid chromatography-tandem mass spectrometry, riboflavin status by eGSH reductase activity coefficient (EGRAC) as well as plasma total cysteine (Cys) and total homocysteine (Hcy), vitamin C by high-performance liquid chromatography, and C-reactive protein (CRP) by standard method. Estimated dietary protein and energy intakes were determined by 3-day food records, and nutritional status was assessed by subjective global assessment (SGA). RESULTS: There were no significant differences among groups in eGSH concentration, EGRAC, dietary protein intake, and SGA score. SHD patients had significantly higher plasma Cys (P < .001) and Hcy compared with HNHD and HC groups (P = .048). Vitamin C was significantly lower (P = .01) and CRP significantly higher (P = .048) in both HD groups compared with HC. CONCLUSION: eGSH concentration appears to be unaffected by dialysis dose in well-nourished HD patients.