Effects of a low and a high dietary LA/ALA ratio on long-chain PUFA concentrations in red blood cells.

There is a debate about the optimal dietary ratio of the parent n6 fatty acid linoleic acid (LA) and n3 fatty acid alpha-linolenic acid (ALA) to promote an efficient conversion of ALA to EPA and DHA, which have implications for human health. The aim of the present study was to compare the effects of a low-LA/high-ALA (loLA/hiALA) diet with a high-LA/low-ALA (hiLA/loALA) diet on fatty acid concentrations in red blood cells (RBCs). Fifteen omnivore healthy men (mean age 26.1 ± 4.5 years) with a low initial EPA/DHA status (sum (∑) EPA + DHA% of total fatty acids in RBC at baseline: 4.03 ± 0.17) received both diets for two weeks with a nine-week wash-out phase in between. Fatty acid intake of the subjects was tightly controlled. Concentrations [µg mL-1] and relative amounts [% of total fatty acids] of fatty acids in RBCs were analyzed at baseline (day 0), day 7 and 14 by means of GC-FID. The dietary LA/ALA ratios were 0.56 ± 0.27 : 1 and 25.6 ± 2.41 : 1 and led to significantly different changes of ALA, LA, EPA and ∑EPA + DHA concentrations in RBCs. In the course of the loLA/hiALA diet ALA and EPA concentrations and relative amounts of ∑EPA + DHA increased, whereas LA concentrations decreased. The DHA concentration was unaffected. The hiLA/loALA diet led to slightly decreased EPA concentrations, while all other fatty acid concentrations remained constant. Compared to our previous study, where we simply increased the ALA intake, our results show that ALA supplementation combined with a reduced LA intake (loLA/hiALA diet) more efficiently enhanced EPA blood concentrations. The absence of changes in the PUFA pattern in consequence of a LA/ALA ratio of 25.6 ± 2.41 : 1 suggests that the high LA/ALA ratio of the Western diet already leads to a saturation and a further increase of the ratio does not affect the PUFA pattern.

Effects of a 12-week high-α-linolenic acid intervention on EPA and DHA concentrations in red blood cells and plasma oxylipin pattern in subjects with a low EPA and DHA status.

The essential omega-3 fatty acid alpha-linolenic acid (ALA, 18:3n3) can be converted into EPA and DHA. The aim of the present study was to determine the effect of a high-ALA diet on EPA and DHA levels in red blood cells (RBCs) and their oxylipins in the plasma of subjects with a low EPA and DHA status. Fatty acid concentrations [µg mL-1] and relative amounts [% of total fatty acids] in the RBCs of 19 healthy men (mean age 26.4 ± 4.6 years) were analyzed by means of GC-FID. Free plasma oxylipin concentrations were determined by LC-MS based targeted metabolomics. Samples were collected and analyzed at baseline (week 0) and after 1 (week 1), 3 (week 3), 6 (week 6), and 12 (week 12) weeks of high dietary ALA intake (14.0 ± 0.45 g day-1). ALA concentrations significantly (p < 0.001) increased from 1.44 ± 0.10 (week 0) to 4.65 ± 0.22 (week 1), 5.47 ± 0.23 (week 3), 6.25 ± 0.24 (week 6), and 5.80 ± 0.28 (week 12) µg mL-1. EPA concentrations increased from 6.13 ± 0.51 (week 0) to 7.33 ± 0.33 (week 1), 8.38 ± 0.42 (p = 0.021, week 3), 10.9 ± 0.67 (p < 0.001, week 6), and 11.0 ± 0.64 (p < 0.001, week 12) µg mL-1. DHA concentrations unexpectedly decreased from 41.0 ± 1.93 (week 0) to 37.0 ± 1.32 (week 1), 36.1 ± 1.37 (week 3), 35.1 ± 1.06 (p = 0.010, week 6), and 30.4 ± 1.09 (p < 0.001, week 12) µg mL-1. Relative ΣEPA + DHA amounts were unchanged during the intervention (week 0: 4.63 ± 0.19, week 1: 4.67 ± 0.16, week 3: 4.61 ± 0.13, week 6: 4.73 ± 0.15, week 12: 4.52 ± 0.11). ALA- and EPA-derived hydroxy- and dihydroxy-PUFA increased similarly to their PUFA precursors, although in the case of ALA-derived oxylipins, the concentrations increased less rapidly and to a lesser extent compared to the concentrations of their precursor FA. LA-derived oxylipins remained unchanged and arachidonic acid and DHA oxylipin concentrations were not significantly changed. Our results confirm that the intake of ALA is not a sufficient source for the increase of EPA + DHA in subjects on a Western diet. Specifically, a high-ALA diet results in increased EPA and declined DHA concentrations. However, the changes effectively balance each other out so that ΣEPA + DHA in RBCs - which is an established marker for health protective effects of omega-3-PUFA - remains constant. The PUFA levels in RBCs reflect the concentration and its changes in plasma hydroxy- and dihydroxy-PUFA concentrations for ALA and EPA.

Effect of DHA supplementation on oxylipin levels in plasma and immune cell stimulated blood.

INTRODUCTION: EPA and DHA cause different physiological effects, which are in many cases mediated via their oxidative metabolites (oxylipins). However, metabolism studies investigating the effect of either EPA or DHA on comprehensive oxylipin patterns are lacking. MATERIAL AND METHODS: The short and long term (1, 3, 6, and 12 week) effect of 1076mg/d DHA (free of EPA) on free (unesterified) oxylipin concentrations in plasma and lipopolysacharid (LPS) stimulated blood of 12 healthy men (mean age 25.1 ± 1.5 years) was investigated. RESULTS: After DHA supplementation, plasma levels of all DHA-oxylipins (HDHAs, EpDPEs, DiHDPEs) significantly increased (up to 600%) in a time-dependent fashion. Oxylipins of EPA and arachidonic acid (AA) were also affected. Whereas a slight increase in several EPA-derived hydroxy-FAs (including the RvE1 precursor 18-HEPE) and dihydroxy-FAs was observed after DHA supplementation, a trend to a slight decline in AA-derived oxylipin levels was found. In LPS stimulated blood, it is shown that DHA supplementation significantly reduces the ability of immune cells to form AA-derived COX (TXB2 and PGB2) and 12-LOX (12-HETE) eicosanoids. While no increase in EPA COX metabolites was found, n-3 PUFA 12-LOX metabolites of EPA (12-HEPE) and DHA (14-HDHA) were highly induced. CONCLUSION: We demonstrated that DHA supplementation causes a time-dependent shift in the entire oxylipin profile suggesting a cross-linked metabolism of PUFAs and subsequent formation of oxygenated lipid mediators.

Effects of docosahexaenoic acid supplementation on PUFA levels in red blood cells and plasma.

INTRODUCTION: Polyunsaturated fatty acids (PUFA) are metabolized in a complex network of elongation, desaturation and beta oxidation. MATERIAL AND METHODS: The short (1 and 3 wk), and long term (6 and 12 wk) effect of 1076mg/d docosahexaenoic acid (DHA, free of eicosapentaenoic acid (EPA)) on (absolute) PUFA concentrations in plasma and red blood cells (RBC) of 12 healthy men (mean age 25.1±1.5 years) was investigated. RESULTS: RBC DHA concentrations significantly (p<0.001) increased from 28±1.6µg/mL to 38±2.0µg/mL (wk 1), 52±3.3µg/mL (wk 3), 68±2.6µg/mL (wk 6), and 79±3.5µg/mL (wk 12). Arachidonic acid (AA) concentrations declined in response to DHA treatment, while the effect was more pronounced in plasma (wk 0: 183±9.9µg/mL, wk 12: 139±8.0µg/mL, -24%, p<0.001) compared to RBC (wk 0: 130±3.7µg/mL, wk 12: 108±4.0µg/mL, -16%, p=0.001). Furthermore, an increase of EPA concentrations in plasma (wk 0: 15±1.5µg/mL, wk 1:19±1.6µg/mL, wk 3: 27±2.3µg/mL, wk 6: 23±1.2µg/mL, wk 12: 25±1.7µg/mL, p<0.001) and RBC (wk 0: 4.7±0.33µg/mL, wk 1: 6.7±1.3µg/mL, wk 3: 8.0±0.66µg/mL, wk 6: 6.9±0.44µg/mL, wk 12: 6.7±0.45µg/mL, n.s.) was observed suggesting a retroconversion of DHA to EPA. CONCLUSION: Based on PUFA concentrations we showed that DHA supplementation results in increased EPA levels, whereas it is not known if this impacts the formation of EPA-derived lipid mediators. Furthermore, shifts in the entire PUFA pattern after supplementation of EPA or DHA should be taken into account when discussing differential physiological effects of EPA and DHA.

Glycemic index and microstructure analysis of a newly developed fiber enriched cookie.

A diet with a high glycemic index (GI) is associated with an elevated risk for obesity or type 2 diabetes. We investigated the GI of a newly-developed fiber enriched cookie and characterized the microstructure of ingredients used. In a study with 26 non-diabetic healthy volunteers it was shown that the fiber enriched cookie has a GI of 58.9 in relation to white bread as reference. Using a conversion factor of 1.4, the GI of the fiber enriched cookie in relation to a glucose-solution is 42.0 and can be classified as a low-GI food. Postprandial insulin concentration was significantly lower after consumption of fiber enriched cookies compared to white bread. Glucose release after in vitro digestion was significantly lower from fiber enriched cookies compared to other cookies tested. In addition to its high percentage of fiber, the cookies' low GI can be attributed to the limited gelatinization potential of the starch granules found in the ingredients used. Using confocal laser scanning microscopy it is shown that starch granule surface area of whole grain barley flour, spelt flour and oat flakes bears cluster-shaped protein-NSPS complexes that preferentially absorb water in conditions of water shortage and thereby prevent starch gelatinization.