Inhibitory Effects of Saponin-Rich Extracts from Pouteria cambodiana against Digestive Enzymes α-Glucosidase and Pancreatic Lipase.

Pouteria cambodiana is a perennial plant that has a wide distribution in tropical regions. It is commonly referred to as 'Nom-nang' in the northern region of Thailand. The bark of this plant has been used for the purpose of promoting lactation among breastfeeding mothers. Moreover, P. cambodiana bark has a high nutraceutical potential due to the presence of saponins, which are secondary metabolites. The purpose of this study was to determine the optimal conditions for ultrasound-assisted extraction (UAE) of saponins from the bark of P. cambodiana and to assess the in vitro inhibitory activities of saponin-rich extracts. The most effective extraction conditions involved a temperature of 50 °C and a 50% concentration level of ethanol as the solvent, which allowed the extraction of saponin at a concentration of 36.04 mg/g. Saponin-rich extracts and their hydrolysates from P. cambodiana bark were evaluated for their ability to inhibit α-glucosidase and pancreatic lipase. The IC50 values for saponin- and sapogenin-rich extracts inhibiting α-glucosidase were 0.10 and 2.98 mg/mL, respectively. Non-hydrolysed extracts also had a stronger inhibitory effect than acarbose. In the case of pancreatic lipase, only the hydrolysed extracts exhibited inhibitory effects on pancreatic lipase (IC50 of 7.60 mg/mL). Thus, P. cambodiana bark may be an applicable natural resource for preparing ingredients for functional products with inhibitory activity against α-glucosidase and pancreatic lipase. The phenolic contents, saponin contents, and antioxidant activities of the dried extract stored at a low temperature of 25 °C for 2 months showed the best stability, with more than 90% retention.

Usual intake of one-carbon metabolism nutrients in a young adult population aged 19-30 years: a cross-sectional study.

One-carbon nutrients play an important role in epigenetic mechanisms and cellular methylation reactions. Inadequate intake of these nutrients is linked to metabolic perturbations, yet the current intake levels of these nutrients have rarely been studied in Asia. This cross-sectional study surveyed the usual dietary intake of one-carbon nutrients (folate, choline and vitamins B2, B6 and B12) among Thai university students aged 19-30 years (n 246). Socioeconomic background, health information, anthropometric data and 24-h dietary recall data were collected. The long-term usual intake was estimated using the multiple-source method. The average usual intake levels for men and women were (mean ± sd) 1⋅85 ± 0⋅95 and 2⋅42 ± 8⋅7 mg/d of vitamin B2, 1⋅96 ± 1⋅0 and 2⋅49 ± 8⋅7 mg/d of vitamin B6, 6⋅20 ± 9⋅5 and 6⋅28 ± 12 µg/d of vitamin B12, 195 ± 154 and 155 ± 101 µg dietary folate equivalent/d of folate, 418 ± 191 and 337 ± 164 mg/d of choline, respectively. Effect modification by sex was observed for vitamin B2 (P-interaction = 0⋅002) and choline (P-interaction = 0⋅02), where every 1 mg increase in vitamin B2 and 100 mg increase in choline intake were associated with a 2⋅07 (P = 0⋅01) and 0⋅81 kg/m2 (P = 0⋅04) lower BMI, respectively, in men. The study results suggest that Thai young adults meet the recommended levels for vitamins B2, B6 and B12. The majority of participants had inadequate folate intake and did not achieve recommended intake levels for choline. The study was approved by the Ethics Committee at the Faculty of Medicine, Chiang Mai University. This trial was registered at www.thaiclinicaltrials.gov (TCTR20210420007).

The shortest innovative process for enhancing the S-allylcysteine content and antioxidant activity of black and golden garlic.

Black garlic is a type of heat-treated garlic for which the traditional process is extremely simple yet time-consuming, taking more than one month. The purpose of this research was to reduce the processing time of black garlic while maintaining a high level of S-allylcysteine (SAC), a black garlic quality indicator. The fresh garlic was pre-treated with CaCl2 and frozen before being further incubated at two different temperatures (60 and 80 °C) with a relative humidity of 65% and 80% RH. Results showed that sequential pre-treatment and incubation at 80 °C and 80% RH for 1 week yielded 874.26 mg of SAC/100 g dry weight with an antioxidant activity of 5390 and 25,421 mg Trolox/100 g for DPPH and ABTS assays, respectively. This process shortened the processing time of black garlic by about 4-times. The batch processed at 60 °C and 65% RH for 1 week provided the highest SAC content of about 1772 mg/100 g dry weight, which was 2-times higher than in incubation at 80 °C and 80% RH for 1 week. The colour of this garlic was golden, so we call this new processed garlic product "golden garlic".

Optimization of Ultrasonic-Assisted Bioactive Compound Extraction from Green Soybean (Glycine max L.) and the Effect of Drying Methods and Storage Conditions on Procyanidin Extract.

Green soybean (Glycine max L.) seeds (GSS) are rich in various antioxidants and phytonutrients that are linked to various health benefits. Ultrasound-assisted extraction (UAE) technology was used for extracting the effective components from GSS. A response surface method (RSM) was used to examine the influence of liquid-to-solid ratio and extraction temperature on the bioactive compounds and antioxidant characteristics. The optimal conditions were a liquid-to-solid ratio of 25:1 and a UAE temperature of 40 °C. The observed values coincided well with the predicted values under optimal conditions. Additionally, the effects of drying methods on the procyanidins and antioxidant activities of GSS extract were evaluated. The spray-dried GSS extract contained the highest levels of procyanidins (21.4 ± 0.37 mg PC/g), DPPH (199 ± 0.85 µM Trolox eq/g), and FRAP (243 ± 0.26 µM Trolox eq/g). Spray drying could be the most time- and energy-efficient technique for drying the GSS extract. The present study also assessed the effects of storage temperature and time on procyanidins and antioxidant activities in GSS extract powder. Procyanidins were found to degrade more rapidly at 45 °C than at 25 °C and 35 °C. Storage under 25 °C was appropriate for maintaining the procyanidin contents, DPPH, and FRAP activities in the GSS extract powder. This study contributed to the body of knowledge by explaining the preparation of procyanidin extract powder from GSS, which might be employed as a low-cost supply of nutraceutical compounds for the functional food industry and pharmaceutical sector.

Prenatal choline supplementation improves biomarkers of maternal docosahexaenoic acid (DHA) status among pregnant participants consuming supplemental DHA: a randomized controlled trial.

BACKGROUND: Dietary methyl donors (e.g., choline) support the activity of the phosphatidylethanolamine N-methyltransferase (PEMT) pathway, which generates phosphatidylcholine (PC) molecules enriched in DHA that are exported from the liver and made available to extrahepatic tissues. OBJECTIVES: This study investigated the effect of prenatal choline supplementation on biomarkers of DHA status among pregnant participants consuming supplemental DHA. METHODS: Pregnant participants (n = 30) were randomly assigned to receive supplemental choline intakes of 550 mg/d [500 mg/d d0-choline + 50 mg/d deuterium-labeled choline (d9-choline); intervention] or 25 mg/d (25 mg/d d9-choline; control) from gestational week (GW) 12-16 until delivery. All participants received a daily 200-mg DHA supplement and consumed self-selected diets. Fasting blood samples were obtained at baseline, GW 20-24, and GW 28-32; maternal/cord blood was obtained at delivery. Mixed-effects linear models were used to assess the impact of prenatal choline supplementation on maternal and newborn DHA status. RESULTS: Choline supplementation (550 vs. 25 mg/d) did not achieve a statistically significant intervention × time interaction for RBC PC-DHA (P = 0.11); a significant interaction was observed for plasma PC-DHA and RBC total DHA, with choline supplementation yielding higher levels (+32-38% and +8-11%, respectively) at GW 28-32 (P < 0.05) and delivery (P < 0.005). A main effect of choline supplementation on plasma total DHA was also observed (P = 0.018); its interaction with time was not significant (P = 0.068). Compared with controls, the intervention group exhibited higher (P = 0.007; main effect) plasma enrichment of d3-PC (d3-PC/total PC). Moreover, the ratio of d3-PC to d9-PC was higher (+50-67%; P < 0.001) in the choline intervention arm (vs. control) at GW 20-24, GW 28-32, and delivery. CONCLUSIONS: Prenatal choline supplementation improves hepatic DHA export and biomarkers of DHA status by bolstering methyl group supply for PEMT activity among pregnant participants consuming supplemental DHA. This trial is registered at www.clinicaltrials.gov as NCT03194659.

Ultrasonic Extraction of Bioactive Compounds from Green Soybean Pods and Application in Green Soybean Milk Antioxidants Fortification.

Green soybean (Glycine max L.) pods (GSP) are agro-industrial waste from the production of frozen green soybean and milk. These pods contain natural antioxidants and various bioactive compounds that are still underutilized. Polyphenols and flavonoids in GSP were extracted by ultrasound technique and used in the antioxidant fortification of green soybean milk. The ultrasound extraction that yielded the highest total polyphenol content and antioxidant activities was 50% amplitude for 10 min. Response surface methodology was applied to analyze an optimum ultrasonic-assisted extraction (UAE) condition of these variables. The highest desirability was found to be 50% amplitude with an extraction time of 10.5 min. Under these conditions, the experimental total phenolic content, total flavonoid content, and antioxidant activity were well matched with the predicted values (R2 > 0.70). Fortification of the GSP extracts (1-3% v/v) in green soybean milk resulted in higher levels of bioactive compounds and antioxidant activity in a dose-dependent manner. Procyanidins were found to be the main polyphenols in dried GSP crude extracts, which were present at a concentration of 0.72 ± 0.01 mg/100 g. The addition of GSP extracts obtained by using an ultrasound technique to green soybean milk increased its bioactive compound content, especially procyanidins, as well as its antioxidant activity.

Choline metabolome response to prenatal choline supplementation across pregnancy: A randomized controlled trial.

Pregnancy places a unique stress upon choline metabolism, requiring adaptations to support both maternal and fetal requirements. The impact of pregnancy and prenatal choline supplementation on choline and its metabolome in free-living, healthy adults is relatively uncharacterized. This study investigated the effect of prenatal choline supplementation on maternal and fetal biomarkers of choline metabolism among free-living pregnant persons consuming self-selected diets. Participants were randomized to supplemental choline (as choline chloride) intakes of 550 mg/d (500 mg/d d0-choline + 50 mg/d methyl-d9-choline; intervention) or 25 mg/d d9-choline (control) from gestational week (GW) 12-16 until Delivery. Fasting blood and 24-h urine samples were obtained at study Visit 1 (GW 12-16), Visit 2 (GW 20-24), and Visit 3 (GW 28-32). At Delivery, maternal and cord blood and placental tissue samples were collected. Participants randomized to 550 (vs. 25) mg supplemental choline/d achieved higher (p < .05) plasma concentrations of free choline, betaine, dimethylglycine, phosphatidylcholine (PC), and sphingomyelin at one or more study timepoint. Betaine was most responsive to prenatal choline supplementation with increases (p ≤ .001) in maternal plasma observed at Visit 2-Delivery (relative to Visit 1 and control), as well as in the placenta and cord plasma. Notably, greater plasma enrichments of d3-PC and LDL-C were observed in the intervention (vs. control) group, indicating enhanced PC synthesis through the de novo phosphatidylethanolamine N-methyltransferase pathway and lipid export. Overall, these data show that prenatal choline supplementation profoundly alters the choline metabolome, supporting pregnancy-related metabolic adaptations and revealing biomarkers for use in nutritional assessment and monitoring during pregnancy.

Associations of choline intake with hypertension and blood pressure among older adults in cross-sectional 2011-2014 National Health and Nutrition Examination Survey (NHANES) differ by BMI and comorbidity status.

Blood pressure (BP) is a known cardiovascular risk factor that is hypothesised to be inversely related to choline intake. A previous study suggested that this association may be more apparent in older adults and may differ according to demographic and health characteristics. The primary study objectives are to investigate the cross-sectional associations of total choline intake with BP (n 843) and prevalent hypertension (n 2113) among USA adults aged ≥ 65 years using the sample from the 2011 to 2014 National Health and Nutrition Examination Survey. Logistic and multiple linear regression models for complex surveys were employed for hypertension status and BP, respectively. Effect modification by sex, race, BMI and comorbidity status were separately investigated using an interaction term. Choline intake interacted with BMI (P-interaction = 0·04) such that choline intake tended to be associated with lower odds of hypertension among people with BMI < 18·5 kg/m2 (OR (95 % CI): 0·64 (0·4, 1·00); P = 0·052). Choline intake was not associated with systolic BP (mean ± sem change per 100 mg of choline: -1·03 ± 0·74 mmHg; P = 0·16). In contrast, its relation to diastolic BP differed by cardiovascular comorbidity (P-interaction = 0·03) with a non-significant (P = 0·13) negative direction of association observed among those who were free of comorbidities and a non-significant (P = 0·26) positive direction observed among those with comorbidities. Collectively, these results suggested that the associations of choline intake with BP levels and hypertension risk among older adults are dependent on other risk factors.

Validation of mathematical model with phosphate activation effect by batch (R)-phenylacetylcarbinol biotransformation process utilizing Candida tropicalis pyruvate decarboxylase in phosphate buffer.

The (R)-phenylacetylcarbinol (PAC) batch biotransformation kinetics for partially purified Candida tropicalis TISTR 5350 pyruvate decarboxylase (PDC) were determined to validate a comprehensive mathematical model in 250 mL scale with 250 mM phosphate buffer/pH 7.0. PDC could convert initial 100/120 mM benzaldehyde/pyruvate substrates to the statistical significantly highest (p ≤ 0.05) maximum PAC concentration (95.8 ± 0.1 mM) and production rate (0.639 ± 0.001 mM min-1). A parameter search strategy aimed at minimizing overall residual sum of square (RSST) based on a system of six ordinary differential equations was applied to PAC biotransformation profiles with initial benzaldehyde/pyruvate concentration of 100/120 and 30/36 mM. Ten important biotransformation kinetic parameters were then elucidated including the zeroth order activation rate constant due to phosphate buffer species (ka) of (9.38 ± < 0.01) × 10-6% relative PDC activity min-1 mM-1. The validation of this model to independent biotransformation kinetics with initial benzaldehyde/pyruvate concentration of 50/60 mM resulted in relatively good fitting with RSST, mean sum of square error (MSE), and coefficient of determination (R2) values of 662, 17.4, and 0.9863, respectively.

Prenatal Nutritional Intervention Reduces Autistic-Like Behavior Rates Among Mthfr-Deficient Mice.

The causes and contributing factors of autism spectrum disorders (ASD) are poorly understood. One gene associated with increased risk for ASD is methylenetetrahydrofolate-reductase (MTHFR), which encodes a key enzyme in one carbon (C1) metabolism. The MTHFR 677C > T polymorphism reduces the efficiency of methyl group production with possible adverse downstream effects on gene expression. In this study, the effects of prenatal and/or postnatal diets enriched in C1 nutrients on ASD-like behavior were evaluated in Mthfr-deficient mice. Differences in intermediate pathways between the mice with and without ASD-like behaviors were tested. The findings indicate that maternal and offspring Mthfr deficiency increased the risk for an ASD-like phenotype in the offspring. The risk of ASD-like behavior was reduced in Mthfr-deficient mice supplemented with C1 nutrients prenatally. Specifically, among offspring of Mthfr+/- dams, prenatal diet supplementation was protective against ASD-like symptomatic behavior compared to the control diet with an odds ratio of 0.18 (CI:0.035, 0.970). Changes in major C1 metabolites, such as the ratios between betaine/choline and SAM/SAH in the cerebral-cortex, were associated with ASD-like behavior. Symptomatic mice presenting ASD-like behavior showed decreased levels of GABA pathway proteins such as GAD65/67 and VGAT and altered ratios of the glutamate receptor subunits GluR1/GluR2 in males and NR2A/NR2B in females. The altered ratios, in turn, favor receptor subunits with higher sensitivity to neuronal activity. Our study suggests that MTHFR deficiency can increase the risk of ASD-like behavior in mice and that prenatal dietary intervention focused on MTHFR genotypes can reduce the risk of ASD-like behavior.

Relation of choline intake with blood pressure in the National Health and Nutrition Examination Survey 2007-2010.

BACKGROUND: Dietary choline is a precursor of trimethylamine N-oxide (TMAO), a metabolite that has been associated with an increased risk of cardiovascular disease. The mechanism underlying this association is unknown, but may include TMAO effects on blood pressure (BP). OBJECTIVES: This study assessed the association of choline intake with hypertension and BP in US adults through the use of NHANES 2007-2010 data. METHODS: This cross-sectional study was conducted in nonpregnant individuals aged ≥20 y. Choline intake was assessed with the use of two 24-h recalls. Outcomes were BP and hypertension status, which was assessed through the use of questionnaires and BP measurements. Modifying factors (e.g., sex, race/ethnicity) and dietary compared with supplemental sources of choline intake were also investigated. RESULTS: The associations of total (dietary + supplemental) and dietary choline intake with the prevalence odds of hypertension differed by sex (n = 9227; P-interaction = 0.04 and 0.03, respectively). In women, both total and dietary choline intake tended to be inversely associated with hypertension (n = 4748; prevalence OR per 100 mg of choline intake: 0.89; 95% CI: 0.77, 1.02; P < 0.10 for both total and dietary choline). No association was observed in men (n = 4479; P = 0.54 and 0.49 for total choline and dietary choline, respectively). Use of choline supplements was inversely associated with hypertension in both sexes (user compared with nonuser; OR: 0.68; 95% CI: 0.49, 0.92; P = 0.01). There was little to no association of total, dietary, or supplemental choline intake with systolic or diastolic BP (n = 6,554; the mean ± SEM change in BP associated with a 100-mg difference in total choline was -0.26 ± 0.22 mm Hg for systolic BP and -0.29 ± 0.19 mm Hg for diastolic BP). CONCLUSIONS: Cross-sectional NHANES data do not support the hypothesis of a positive association between choline intake and BP.

The metabolic fate of isotopically labeled trimethylamine-N-oxide (TMAO) in humans.

Trimethylamine-N-oxide (TMAO) is associated with chronic disease risk. However, little is known about the metabolic fate of dietary TMAO. This study sought to quantitatively elucidate the metabolic fate of orally consumed TMAO in humans. As part of a crossover feeding study, healthy young men (n=40) consumed 50-mg deuterium-labeled methyl d9-TMAO (d9-TMAO), and enrichments of TMAO and its derivatives were measured in blood for 6 h, urine and stool, as well as skeletal muscle in a subset of men (n=6). Plasma d9-TMAO was detected as early as 15 min, increased until 1 h and remained elevated through the 6-h period. TMAO exhibited an estimated turnover time of 5.3 h, and ~96% of the dose was eliminated in urine by 24 h, mainly as d9-TMAO. No d9-TMAO was detected in feces. Notably, d9-TMAO and d9-trimethylamine were detected in skeletal muscle (n=6) at 6 h, and the enrichment ratio of d9-TMAO to d9-trimethylamine was influenced by a genetic variant in flavin-containing monooxygenase isoform 3 (FMO3 G472A). These results suggest that the absorption of orally consumed TMAO is near complete and does not require processing by gut microbes. TMAO exhibits fast turnover in the circulation with the majority being eliminated in urine within 24 h. A small portion of the dose, however, is taken up by extrahepatic tissue in a manner that appears to be under the influence of FMO3 G472A polymorphism. This trial was registered at clinicaltrials.gov as NCT02558673.

Trimethylamine-N-oxide (TMAO) response to animal source foods varies among healthy young men and is influenced by their gut microbiota composition: A randomized controlled trial.

SCOPE: Trimethylamine-N-oxide (TMAO), a metabolite linked to the gut microbiota, is associated with excess risk of heart disease. We hypothesized that (i) TMAO response to animal source foods would vary among healthy men and (ii) this response would be modified by their gut microbiome. METHODS AND RESULTS: A crossover feeding trial in healthy young men (n = 40) was conducted with meals containing TMAO (fish), its dietary precursors, choline (eggs) and carnitine (beef), and a fruit control. Fish yielded higher circulating and urinary concentrations of TMAO (46-62 times; p < 0.0001), trimethylamine (8-14 times; p < 0.0001), and dimethylamine (4-6-times; P<0.0001) than eggs, beef, or the fruit control. Circulating TMAO concentrations were increased within 15 min of fish consumption, suggesting that dietary TMAO can be absorbed without processing by gut microbes. Analysis of 16S rRNA genes indicated that high-TMAO producers (≥20% increase in urinary TMAO in response to eggs and beef) had more Firmicutes than Bacteroidetes (p = 0.04) and less gut microbiota diversity (p = 0.03). CONCLUSION: Consumption of fish yielded substantially greater increases in circulating TMAO than eggs or beef. The higher Firmicutes to Bacteroidetes enrichment among men exhibiting a greater response to dietary TMAO precursor intake indicates that TMAO production is a function of individual differences in the gut microbiome.

Choline intakes exceeding recommendations during human lactation improve breast milk choline content by increasing PEMT pathway metabolites.

Demand for the vital nutrient choline is high during lactation; however, few studies have examined choline metabolism and requirements in this reproductive state. The present study sought to discern the effects of lactation and varied choline intake on maternal biomarkers of choline metabolism and breast milk choline content. Lactating (n=28) and control (n=21) women were randomized to 480 or 930 mg choline/day for 10-12 weeks as part of a controlled feeding study. During the last 4-6 weeks, 20% of the total choline intake was provided as an isotopically labeled choline tracer (methyl-d9-choline). Blood, urine and breast milk samples were collected for choline metabolite quantification, enrichment measurements, and gene expression analysis of choline metabolic genes. Lactating (vs. control) women exhibited higher (P < .001) plasma choline concentrations but lower (P ≤ .002) urinary excretion of choline metabolites, decreased use of choline as a methyl donor (e.g., lower enrichment of d6-dimethylglycine, P ≤ .08) and lower (P ≤ .02) leukocyte expression of most choline-metabolizing genes. A higher choline intake during lactation differentially influenced breast milk d9- vs. d3-choline metabolite enrichment. Increases (P ≤ .03) were detected among the d3-metabolites, which are generated endogenously via the hepatic phosphatidylethanolamine N-methyltransferase (PEMT), but not among the d9-metabolites generated from intact exogenous choline. These data suggest that lactation induces metabolic adaptations that increase the supply of intact choline to the mammary epithelium, and that extra maternal choline enhances breast milk choline content by increasing supply of PEMT-derived choline metabolites. This trial was registered at clinicaltrials.gov as NCT01127022.