Low-dose daily folic acid (400 µg) supplementation does not affect regulation of folate transporters found present throughout the terminal ileum and colon of humans: a randomized clinical trial.

BACKGROUND: Folic acid supplementation during the periconceptional period reduces the risk of neural tube defects in infants, but concern over chronic folic acid exposure remains. An improved understanding of folate absorption may clarify potential risks. Folate transporters have been characterized in the small intestine, but less so in the colon of healthy, free-living humans. The impact of folic acid fortification or supplementation on regulation of these transporters along the intestinal tract is unknown. OBJECTIVE: The objective was to characterize expression of folate transporters/receptor (FT/R) and folate hydrolase, glutamate carboxypeptidase II (GCPII), from the terminal ileum and throughout the colon of adults and assess the impact of supplemental folic acid. METHODS: In this 16-wk open-labeled randomized clinical trial, adults consumed a low folic acid-containing diet, a folate-free multivitamin, and either a 400 µg folic acid supplement or no folic acid supplement. Dietary intakes and blood were assessed at baseline, 8 wk, and 16 wk (time of colonoscopy). Messenger RNA (mRNA) expression and protein expression of FT/R and GCPII were assessed in the terminal ileum, cecum, and ascending and descending colon. RESULTS: Among 24 randomly assigned subjects, no differences in dietary folate intake or blood folate were observed at baseline. Mean ± SD red blood cell folate at 16 wk was 1765 ± 426 and 911 ± 242 nmol/L in the 400 and 0 µg folic acid group, respectively (P < 0.0001). Reduced folate carrier, proton-coupled folate transporter, and folate-receptor alpha expression were detected in the terminal ileum and colon, as were efflux transporters of breast cancer resistance protein and multidrug resistance protein-3. Other than a higher mRNA expression of FR-alpha and GCPII in the 400 µg supplement group in the ascending colon, no treatment differences were observed (P < 0.02). CONCLUSIONS: Folate transporters are present throughout the terminal ileum and colon; there is little evidence that a low dose of folic acid supplementation affects colonic absorption. This trial was registered at clinicaltrials.gov as NCT03421483.

Term Infants Fed Exclusively With Donor Milk May Require Vitamin C Supplementation.

BACKGROUND: When there is insufficient mother's milk for preterm infants, fortified human donor milk (DM) is the preferred supplement. Recently, there is growing interest in providing DM to term infants. Aside from vitamin D, mother's milk is a complete source of nutrition for term infants. It is unknown whether supplementation of micronutrients is required for term infants exclusively fed DM, particularly for nutrients affected by heat processing, such as vitamin C. The objective of this study was to determine the total vitamin C content in DM and whether it would be adequate for an infant exclusively fed DM. METHODS: DM samples (n = 56) were collected at a Canadian milk bank from April to August 2018. Vitamin C concentration was determined by high-performance liquid chromatography. RESULTS: DM samples had a vitamin C concentration of 17.7 ± 9.8 mg/L (mean ± SD) and were variable, ranging from 1.9 to 43.2 mg/L. Using these values and assuming an exclusive DM consumption of 780 mL/day, the estimated vitamin C intake would be 13.8 ± 8.6 mg (mean ± SD), falling below the adequate intake of 40 mg/day for infants (0-6 months old). CONCLUSION: Vitamin C supplementation is required for all infants if DM is the sole source of nutrition. Future studies should investigate other heat- and light-sensitive nutrients.

High Hydrostatic Pressure Processing Better Preserves the Nutrient and Bioactive Compound Composition of Human Donor Milk.

BACKGROUND: When mother's milk is insufficient, pasteurized human donor milk (DM) is the recommended supplement for hospitalized very-low-birth-weight infants. The current method of pasteurization (Holder, 62.5°C, 30 min) negatively affects heat-sensitive nutrients and bioactive proteins. OBJECTIVES: Objectives of this study were to compare changes in DM composition after thermal pasteurization (Holder and flash-heating) and nonthermal methods [UV-C irradiation and high hydrostatic pressure (HHP)]. We hypothesized that nonthermal techniques would result in fewer changes to composition. METHODS: Holder, flash-heating (brought to boil), UV-C irradiation (250 nm, 25 min), and HHP (500 MPa, 8 min) were studied. Pools of milk from 17 women known to contain bacteria at >5 × 107 colony forming units (CFU)/L were collected from the Rogers Hixon Ontario Human Milk Bank and underwent each pasteurization technique. Macronutrients, heat-sensitive micronutrients (vitamin C, folate), and bioactive components [bile-salt-stimulated lipase (BSSL), lysozyme, lactoferrin] were measured in raw and pools of pasteurized milk. Milk was cultured to determine how well each technique produced a culture negative result (detection limit <1 × 103 CFU/L). RESULTS: Folate was reduced by 24-27% after Holder, flash-heating, and UV-C (P < 0.05); no reduction was observed after HHP. All pasteurization methods reduced vitamin C (60-75%, P < 0.001). BSSL was abolished after Holder and flash-heating (P < 0.001), reduced after UV-C (48%, P < 0.001), but unaffected by HHP. Lysozyme activity was reduced after flash-heating (44%) and UV-C (74%, P < 0.004) but unaffected by Holder or HHP. Lactoferrin was reduced by all methods (P < 0.02) but most severely by flash-heating (74%) and least severely by HHP (25%). Holder and UV-C reduced lactoferrin by ∼48%. All pasteurization methods reduced the number of culture positive DM samples (P < 0.001). CONCLUSIONS: HHP better preserves human milk composition than Holder pasteurization. Future research on the feasibility of HHP for pasteurizing human milk is warranted because its implementation may improve the nutritional status and health of DM-fed infants.

High concentrations of folate and unmetabolized folic acid in a cohort of pregnant Canadian women and umbilical cord blood.

BACKGROUND: Mandatory fortification, prevalent supplement use, and public health guidelines recommending periconceptional supplementation have increased folic acid intakes in North American pregnant women. However, the effects of increased folic acid intakes during pregnancy on maternal and cord blood folate concentrations have not been well established. OBJECTIVES: In this prospective study, we determined maternal and cord blood concentrations of folate and unmetabolized folic acid (UMFA) in a cohort of pregnant Canadian women and their newborns and examined the effect of maternal intakes of folate and folic acid and fetal genetic variants in folate metabolism on folate status. DESIGN: Folate and folic acid intakes of 368 Canadian pregnant women were assessed in early (0-16 wk) and late (23-37 wk) pregnancy. Blood concentrations of folate and UMFA were measured with the use of immunoassays and liquid chromatography-mass spectrometry, respectively, in maternal samples in early pregnancy (12-16 wk), at delivery (28-42 wk), and in cord blood. Four fetal genetic variants of the 5,10-methylenetetrahydrofolate reductase (MTHFR) and dihydrofolate reductase (DHFR) genes were assessed for their association with cord blood concentrations of folate and UMFA. RESULTS: Geometric mean (95% CI) maternal red blood cell (RBC) folate concentrations were 2417 nmol/L (2362, 2472 nmol/L ) and 2793 nmol/L (2721, 2867 nmol/L ) in early pregnancy and at delivery, respectively. The mean (95% CI) cord RBC folate concentration was 2689 nmol/L (2614, 2765 nmol/L). UMFA was detectable in >90% of maternal and cord plasma samples. Although 3 fetal MTHFR and DHFR genetic variants had no effect, the fetal MTHFR 677TT genotype was associated with significantly lower cord serum (P = 0.03) and higher cord RBC (P = 0.02) folate concentrations than those of the wild type. CONCLUSIONS: Notwithstanding differences in assays, maternal and cord RBC folate and plasma UMFA concentrations were higher than previously reported values. Functional ramifications of high folate and UMFA concentrations in maternal and fetal circulation warrant additional investigation because an excess folate status may affect long-term health outcomes of the offspring. This study was registered at www.clinicaltrials.gov as NCT02244684.

The direction of the difference between Canadian and American erythrocyte folate concentrations is dependent on the assay method employed: a comparison of the Canadian Health Measures Survey and National Health and Nutrition Examination Survey.

Fortification of select grain products with folic acid and periconceptional supplementation recommendations in Canada and the USA have improved folate status, and have been associated with a reduced risk of neural tube defects. In the present study, we aimed to conduct a comparison of erythrocyte folate concentrations from the 2007-9 Canadian Health Measures Survey (CHMS) and the 2007-8 US National Health and Nutrition Examination Survey (NHANES). Erythrocyte folate concentration was assessed in participants aged 6-79 years (CHMS, n 5248; NHANES, n 7070). To account for different folate assays employed - Immulite 2000 immunoassay (CHMS) and microbiological assay (NHANES) - a conversion equation was generated (n 152 adults) to adjust the CHMS data. t Tests were used to examine country differences. Median Canadian erythrocyte folate concentrations (method-adjusted) were lower than those of Americans (988 and 1100 nmol/l, respectively), but unadjusted median Canadian erythrocyte folate concentrations were higher (1250 nmol/l). The upper 95% CI boundary of the method-adjusted Canadian erythrocyte folate distribution overlapped that of the American erythrocyte folate concentrations, while the lower 95% CI boundary of the method-adjusted Canadian erythrocyte folate data was below the American distribution. In summary, the fact that erythrocyte folate concentrations were either higher or lower in Canadians compared with Americans, depending on whether an adjustment was made to account for assay differences, suggests that caution must be exercised in evaluating erythrocyte folate data from different countries because analytical methods are not readily comparable. Furthermore, we cannot unequivocally conclude that there are true differences in erythrocyte folate concentrations between the Canadian and American populations in the post-fortification era.

Dietary oligosaccharides increase colonic weight and the amount but not concentration of bacterially synthesized folate in the colon of piglets.

Recently, we demonstrated that the large pool of folate present in the colon of humans can be absorbed. Here, we investigated whether the net amount of folate synthesized by bacteria in the colon of piglets can be modified by feeding prebiotics to alter their microbial milieu. Male piglets (age 5 d, n = 12) were randomized to a milk-based formula containing either 5 g/L inulin + 5 g/L galactooligosaccharides (IN-GOS) or 5 g/L maltodextrin (control). Body weight did not differ between groups during the 28-d feeding intervention. However, the mean weight of colonic tissue (38%) and their contents (238%) was higher in the IN-GOS than in the control group (P = 0.004, P = 0.0001, respectively). Total bacterial load in the colon of piglets fed IN-GOS was 531% greater and the total amount of folate found in the colon contents was 53% greater than that of controls (P = 0.002, P = 0.02, respectively). Indices of blood folate status (plasma and RBC folate and plasma homocysteine) and folate concentrations in liver and kidneys were unaffected. Both groups exhibited low RBC folate (56 ± 23 nmol/L) and elevated homocysteine (24 ± 7 µmol/L) concentrations, evidence of deficiency if present in humans. In conclusion, dietary supplementation with 5 g of inulin + 5 g of galactooligosaccharides increased the weight, bacterial load, and total folate content in the piglet colon; however, these changes were insufficient to modify indices of whole body folate status. Future studies investigating the impact of feeding prebiotics on localized folate status at the level of the colonocyte are warranted.