Long-chain Polyunsaturated Fatty Acid Ratio Research Articles

Infant formula containing large, milk phospholipid-coated lipid droplets and dairy lipids affects cognitive performance at school age.

Breastfeeding has been positively associated with infant and child neurocognitive development and function. Contributing to this effect may be differences between human milk and infant formula in the milk lipid composition and milk fat globule structure. To evaluate the effects of an infant formula mimicking human milk lipid composition and milk fat globule structure on childhood cognitive performance. In a randomized, controlled trial, healthy term infants received until 4 months of age either a Standard infant formula (n = 108) or a Concept infant formula (n = 115) with large, milk phospholipid coated lipid droplets and containing dairy lipids. A breastfed reference group (n = 88) was included. Erythrocyte fatty acid composition was determined at 3 months of age. Neurocognitive function was assessed as exploratory follow-up outcome at 3, 4, and 5 years of age using the Flanker test, Dimensional Change Card Sort (DCCS) test and Picture Sequence Memory test from the National Institutes of Health Toolbox Cognition Battery. Mann-Whitney U test and Fisher exact test were used to compare groups. Erythrocyte omega-6 to -3 long-chain polyunsaturated fatty acid ratio appeared to be lower in the Concept compared to the Standard group (P = 0.025). At age 5, only the Concept group was comparable to the Breastfed group in the highest reached levels on the Flanker test, and the DCCS computed score was higher in the Concept compared to the Standard group (P = 0.021). These outcomes suggest that exposure to an infant formula mimicking human milk lipid composition and milk fat globule structure positively affects child neurocognitive development. Underlying mechanisms may include a different omega-3 fatty acid status during the first months of life. https://onderzoekmetmensen.nl/en/trial/28614, identifier NTR3683 and NTR5538.

HIGH OLEIC SUNFLOWER OIL DECREASES ENDOGENOUS BIOSYNTHESIS OF ENERGY FATTY ACIDS AND INCREASES ENDOGENOUS BIOSYNTHESIS OF ω-3 LONG-CHAIN PUFA

The work shows that Fatty acids of dietary fats provide two main functions in the human and animal body: energy and structural-regulatory, Polyunsaturated fatty acids (PUFA) are the basis of membrane lipids of all cells of the body The structure and functional activity of cells, their resistance to pathogenic factors depends on the ratio of ω-6 / ω-3 PUFA. Аdherence to recommended metabolic energy reserves in poultry feed is essential to optimize feed costs. The use of oils or fats is a common economic practice in modern poultry production. Energy functions are carried out due to the oxidation of energy fatty acids in mitochondria, which include, first of all, palmitic (С16:0), palmitooleic (С16:0), stearic (С18:1) and oleic (С18:1). In addition to providing energy, edible oil can also enhance dietary palatability, reduce dustiness, and increase lipoprotein hydrolysis to promote the production of essential fatty acids. Adipose tissue should be considered not only as a source of various fatty acids, but also as an important endocrine organ that takes an active part in the activity of the immune system. To determine the effect of a diet with high oleic sunflower oil on the content of energy fatty acids (EFA) and long-chain PUFA (LCPUFA) in rat liver lipids. We used high oleic sunflower oil (HOSO) containing 85.5% oleic acid. The rats were fed a fat-free diet (FFD) and diets with 5 or 15% HOSO for 35 days. Lipids were extracted from the liver and separated into three fractions: neutral lipids (NL), phospholipids (PL) and free fatty acids (FFA). The fatty acid composition of lipid fractions was determined by gas chromatography. FFA are the sum of the following acids:С16:0, С16:1, С18:0, С18:1andС18:2. LCPUFA are presented С20:4 ω-6, С20:5 ω-3, С22:5 ω-3and С22:6 ω-3. Most of all, EFA is contained in the NL fraction (89%), then in the PL fraction (79%), and least of all in the FFA fraction (68%). LCPUFA is found most of all in the FFA fraction (20%), then in the PL fraction (13%), and least of all in the NL fraction (2%). In rats that received fat diets, the content of EFA increased in the NL fraction by 2-3%, in the FFA fraction by 5-8%, and did not change in the PL fraction. The content of LCPUFA ω-6 (arachidonic acid) with fat nutrition dose-dependently decreases in the fraction of NL and FFA and does not change in the fraction of PL. On the contrary, the content of ω-3 LCPUFA increases in rats treated with HOSO in all lipid fractions. Also, the ω-6/ω-3 LCPUFA ratio is significantly reduced in rats treated with HOSO. Consumption of HOSO stimulates endogenous biosynthesis of ω-3 LCPUFA.

Associations of Maternal Consumption of Dairy Products during Pregnancy with Perinatal Fatty Acids Profile in the EDEN Cohort Study.

Maternal diet is the main source of fatty acids for developing offspring in-utero and in breastfed infants. Dairy products (DP) are important sources of fat in the European population diet. C15:0 and C17:0 fatty acids have been suggested as biomarkers of dairy fat consumption. This study’s aim is to describe the associations between maternal DP (milk included) consumption during pregnancy and C15:0, C17:0 and polyunsaturated fatty acid (PUFA) levels in perinatal biofluids. Study populations were composed of 1763, 1337 and 879 French mothers from the EDEN (“Étude des Déterminants pre- et post-natals de la santé de l’ENfant”) study, with data on maternal and cord red blood cells’ (RBC) membrane and colostrum, respectively. Associations were assessed using linear regression models adjusted for recruitment center, maternal age, healthy dietary pattern or fish consumption. Greater adherence to a ”cheese” consumption pattern was associated with lower linoleic acid level in colostrum and higher C15:0 and C17:0 levels but in a less consistent manner for C17:0 across biofluids. Greater adherence to “semi-skimmed milk, yogurt” and “reduced-fat DP” patterns was related to higher docosahexaenoic acid and total n-3 PUFA levels and lower n-6/n-3 long-chain PUFA ratio in maternal and cord RBC. Our results suggest that C15:0 could be a good biomarker of maternal dairy fat consumption in perinatal biofluids.

Effect of different dietary arachidonic, eicosapentaenoic, and docosahexaenoic acid content on selected immune parameters in gilthead sea bream (Sparus aurata)

The increasing replacement of fish oil (FO) by vegetable oils (VO) in aquafeeds for marine carnivorous species may lead to modifications of the dietary arachidonic (ARA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) levels and ratios that may compromise the fish immune status. Therefore, this study aimed to assess how different dietary ratios of ARA, EPA, and DHA modulate the immune response of gilthead sea bream juveniles. For that purpose, four isoproteic and isolipidic plant-based diets were formulated to include different dietary Long-Chain-Polyunsaturated Fatty Acids (LC-PUFA) ratios (%DM): Diet A (2.0% ARA: 0.2% EPA: 0.1% DHA); Diet B (1.0% ARA: 0.4% EPA: 0.4% DHA); Diet C (0% ARA: 0.6% EPA: 0.6% DHA); Diet D (0% ARA: 0.3% EPA: 1.5% DHA). Fish were fed the experimental diets for 56 days and then the haematological profile, humoral innate immune parameters, and expression of immune-related genes in the gut were determined. Our results showed that fish fed diet B exhibited an increase in the monocyte numbers, alternative complement activity, and bactericidal activity against Photobacterium damselae compared to the values found in fish fed other diets. Contrarily, peroxidase, antiproteases, and proteases activity and nitric oxide levels were not affected by the dietary treatments. Immune-related gene expression in the distal intestine did not show differences between dietary treatments. Overall, a diet with a balanced n-6 (ARA) and n-3 (EPA+DHA) LC-PUFA ratio seemed to better modulate the fish innate immune response, thus better preparing the fish innate immune system against potential immunological insults.

Changes in peripheral immune populations during pregnancy and modulation by probiotics and \u03c9-3 fatty acids

Allergic diseases have become a major health problem, partly due to reduced microbial stimulation and a decreased dietary ω-3/ω-6 long-chain polyunsaturated fatty acid ratio. Prenatal exposures have been reported to influence allergy development, possibly induced via changes in maternal immune regulation. In a randomized double-blind placebo-controlled multicenter allergy prevention trial (PROOM-3), pregnant women were recruited at gestational week 20, and randomized to four study groups, one receiving both L. reuteri oil drops and ω-3 PUFA capsules (n = 22), the second receiving ω-3 PUFA supplementation and placebo regarding L. reuteri (n = 21), the third receiving L. reuteri and placebo regarding ω-3 PUFA (n = 22) and the fourth group receiving placebo capsules and placebo oil drops (n = 23). In this substudy, supplemental and pregnancy-related effects on maternal peripheral immune cell populations during pregnancy were assessed by flow cytometry immune phenotyping at gestational week 20, 32 and 4 days after delivery. The numbers of activated and regulatory T (Treg) cells (CD45RA− Foxp3++/CD45RA+Foxp3+) were reduced after delivery, with the lowest count in the L. reuteri supplemented group compared with the placebo group 4 days after delivery, while the ω-3 PUFA group did not differ from the placebo group. Several treatment-independent changes were observed during and after pregnancy in lymphocytes (CD4+/8+/19+/56+/45RA+/−), CD14+16+/− monocytes, and in subpopulations of T helper cells (Th) CD4+CD45RA−Tbet+ (Th1) and CD4+CD45RA−RORC+ (Th17) cells. In conclusion, probiotic supplementation to the mother during the second half of pregnancy resulted in immunomodulatory effects among activated and resting Treg cells. Furthermore, several systemic immune modifying effects of pregnancy were observed.

Iron-induced derangement in hepatic Δ-5 and Δ-6 desaturation capacity and fatty acid profile leading to steatosis: Impact on extrahepatic tissues and prevention by antioxidant-rich extra virgin olive oil

The administration of iron induces liver oxidative stress and depletion of long-chain polyunsaturated fatty acids (LCPUFAs), n-6/n-3 LCPUFA ratio enhancement and fat accumulation, which may be prevented by antioxidant-rich extra virgin olive oil (AR-EVOO) supplementation. Male Wistar rats were subjected to a control diet (50 mg iron/kg diet) or iron-rich diet (IRD; 200 mg/kg diet) with alternate AR-EVOO for 21 days. Liver fatty acid (FA) analysis was performed by gas-liquid chromatography (GLC) after lipid extraction and fractionation, besides Δ-5 desaturase (Δ-5 D) and Δ6-D mRNA expression (qPCR) and activity (GLC) measurements. The IRD significantly (p < 0.05) increased hepatic total fat, triacylglycerols, free FA contents and serum transaminases levels, with diminution in those of n-6 and n-3 LCPUFAs, higher n-6/n-3 ratios, lower unsaturation index and Δ5-D and Δ6-D activities, whereas the mRNA expression of both desaturases was enhanced over control values, changes that were prevented by concomitant AR-EVOO supplementation. N-6 and n-3 LCPUFAs were also decreased by IRD in extrahepatic tissues and normalized by AR-EVOO. In conclusion, AR-EVOO supplementation prevents IRD-induced changes in parameters related to liver FA metabolism and steatosis, an effect that may have a significant impact in the treatment of iron-related pathologies or metabolic disorders such as non-alcoholic fatty liver disease.

Methylmercury and long chain polyunsaturated fatty acids are associated with immune dysregulation in young adults from the Seychelles child development study.

BackgroundExposure to the environmental toxicant mercury (Hg) has been associated with immune dysregulation, including autoimmune disease, but few human studies have examined methylmercury (MeHg) exposure from fish consumption. ObjectivesWe examined associations between MeHg exposure and biological markers of autoimmunity and inflammation while adjusting for long chain polyunsaturated fatty acids (LCPUFA). MethodAt age 19 years, hair total Hg (Y19Hg), LCPUFA status, a panel of 13 antinuclear antibodies (ANA), total serum immunoglobulins (Ig) IgG, IgA, and IgM and serum markers of inflammation (IL-1, IL-2, IL-6, IL-10, C-reactive protein (CRP), IFN-γ, TNF-α) were measured in the Seychelles Child Development Study (SCDS) Main Cohort (n = 497). Multivariable regression models investigated the association between Y19Hg and biomarkers, adjusting for prenatal total hair Hg (MatHg) and other relevant covariates, and with and without adjustment for LCPUFA. ResultsWith each 1 ppm increase in Y19Hg (mean 10.23 (SD 6.02) ppm) we observed a 4% increased odds in a positive Combined ANA following adjustment for the n6:n3 LCPUFA ratio (β = 0.036, 95%; CI: 0.001, 0.073). IgM was negatively associated with Y19Hg (β = −0.016, 95%CI: 0.016, −0.002) in models adjusted for n-3, n-6 LCPUFA and when separately adjusted for the n-6:n-3 LCPUFA ratio. No associations were observed with MatHg. Total n-3 LCPUFA status was associated with reduced odds of a positive anti-ribonuclear protein (RNP) A. The n-3 LCPUFA were negatively associated with IL-6, IL-10, CRP, IFN-γ, TNF-α and positively with TNF-α:IL-10. There were positive associations between the n-6:n-3 ratio and IL-6, IL-10, CRP, IFN-γ, TNF-α and a negative association with TNF-α:IL-10. DiscussionThe Y19Hg exposure was associated with higher ANA and lower IgM albeit only following adjustment for the n-3 LCPUFA or the n-6:n-3 LCPUFA ratio. The clinical significance of these findings is unclear, but warrant follow up at an older age to determine any relationship to the onset of autoimmune disease.

Associations between methylmercury, n-3 polyunsaturated fatty acids and antinuclear antibodies in young adults from the Seychelles Child Development Study (SCDS).

AbstractThe association of MeHg exposure through fish consumption on human autoimmunity remains unclear. Fish also contain n-3 long chain polyunsaturated fatty acids (LCPUFA) that are known to regulate inflammation and mitigate autoimmune disease symptoms. We studied the association of low-level exposure to methylmercury (MeHg) through fish consumption in the SCDS. We examined this association at age 19 years in the SCDS Main Cohort (n = 497). We measured MeHg exposure at 3 time points [prenatal, weighted average (6 months to 19 years) and concurrent (19 years) and LCPUFA status and a panel of 13 autoimmune markers at age 19 years. The autoimmune markers included antinuclear antibodies (ANA), anti-dsDNA and anti-RNP, and total (non-specific) immunoglobulins (Ig) IgG, IgA, and IgM. A combined ANA variable was also calculated based on being within or above reference range for any of the ANA markers; 56% of the subjects met this criterion. Multivariable regression models adjusted for prenatal MeHg, sex and waist circumference, with and without adjustment for LCPUFA, were fit for the three MeHg exposure metrics and each immune marker. Mean (SD) prenatal, weighted average and concurrent MeHg was 6.84 (4.55), 7.46 (2.82), and 10.23 (6.02) ppm, respectively. Combined ANA was positively associated with concurrent MeHg following adjustment for the n6:n3 LCPUFA ratio (β = 0.036, 95%; CI: 0.001, 0.073). Prenatal and average MeHg exposures were not significantly associated with any individual ANA. IgM was negatively associated with concurrent (β = -0.016, 95%CI: -0.016, -0.002), and average (β = -0.042, 95%CI: -0.042, -0.009) MeHg exposure in the models adjusted for n-3, n-6 LCPUFA and when separately adjusted for the n6:n3 LCPUFA ratio. Total (19-year) n-3 PUFA status was negatively associated with anti-RNP (β = -20.355, 95%CI: -36.89, -4.34) and IgG (β = -1.384, 95%CI: -2.682, -0.087). Total n-3 LCPUFA was associated with lower markers of autoimmunity. MeHg exposure at 19 years was associated with higher ANA and lower IgM but only following adjustment for LCPUFA. The clinical significance of these findings is unclear and further research is warranted to determine if these associations precede autoimmune disease development.

Ovarian Hormone-Dependent Effects of Dietary Lipids on APP/PS1 Mouse Brain.

The formation of senile plaques through amyloid-β peptide (Aβ) aggregation is a hallmark of Alzheimer’s disease (AD). Irrespective of its actual role in the synaptic alterations and cognitive impairment associated with AD, different therapeutic approaches have been proposed to reduce plaque formation. In rodents, daily intake of omega-3 (n-3) long-chain polyunsaturated fatty acids (LC-PUFAs) is required for neural development, and there is experimental and epidemiological evidence that their inclusion in the diet has positive effects on several neurodegenerative diseases. Similarly, estradiol appears to reduce senile plaque formation in primary mouse cell cultures, human cortical neurons and mouse AD models, and it prevents Aβ toxicity in neural cell lines. We previously showed that differences in dietary n-6/n-3 LC-PUFAs ratios modify the lipid composition in the cerebral cortex of female mice and the levels of amyloid precursor protein (APP) in the brain. These effects depended in part on the presence of circulating estradiol. Here we explored whether this potentially synergistic action between diet and ovarian hormones may influence the progression of amyloidosis in an AD mouse model. Our results show that a diet with high n-3 LC-PUFA content, especially DHA (22:6n-3), reduces the hippocampal accumulation of Aβ1–40, but not amyloid Aβ1–42 in female APPswe/PS1 E9A mice, an effect that was counteracted by the loss of the ovaries and that depended on circulating estradiol. In addition, this interaction between dietary lipids and ovarian function also affects the composition of the brain lipidome as well as the expression of certain neuronal signaling and synaptic proteins. These findings provide new insights into how ovarian hormones and dietary composition affect the brain lipidome and amyloid burden. Furthermore, they strongly suggest that when designing dietary or pharmacological strategies to combat human neurodegenerative diseases, hormonal and metabolic status should be specifically taken into consideration as it may affect the therapeutic response.

Prenatal Omega-6:Omega-3 Ratio and Attention Deficit and Hyperactivity Disorder Symptoms

To evaluate whether higher omega-6:omega-3 (n-6:n-3) long-chain polyunsaturated fatty acid ratio in cord plasma is associated with more symptoms of attention deficit and hyperactivity disorder (ADHD) at 4 and 7years of age. This study was based on a population-based birth cohort in Spain. N-6 arachidonic acid and n-3 eicosapentaenoic and docosahexaenoic acid concentrations were measured in cord plasma. At 4years old, ADHD symptoms were reported by teachers through the ADHD Diagnostic and Statistical Manual of Mental Disorders, 4th ed checklist (n=580). At 7years old, ADHD symptoms were reported by parents through the Conners' Rating Scale-Revised (short form; n=642). The ADHD variable was treated as continuous (score) and as dichotomous (symptom diagnostic criteria). Child and family general characteristics were prospectively collected through questionnaires. We applied pooled zero-inflated negative binomial and logistic regressions adjusted for covariates. A higher omega-6:omega-3 long-chain polyunsaturated fatty acid ratio in cord plasma was associated with a higher ADHD index (incidence rate ratio, 1.13; 95% CI, 1.03, 1.23) at 7years old. The association was not observed at 4years old (incidence rate ratio, 1.04; 95% CI, 0.92-1.18). No associations were found using ADHD symptom diagnostic criteria. High prenatal omega-6:omega-3 long-chain polyunsaturated fatty acid ratio preceded the appearance of subclinical ADHD symptoms during mid-childhood. Our findings suggest that maternal diet during pregnancy may modulate the risk to develop long-term ADHD symptoms in the offspring.

Ovarian Function Modulates the Effects of Long-Chain Polyunsaturated Fatty Acids on the Mouse Cerebral Cortex.

Different dietary ratios of n−6/n−3 long-chain polyunsaturated fatty acids (LC-PUFAs) may alter brain lipid profile, neural activity, and brain cognitive function. To determine whether ovarian hormones influence the effect of diet on the brain, ovariectomized and sham-operated mice continuously treated with placebo or estradiol were fed for 3 months with diets containing low or high n−6/n−3 LC-PUFA ratios. The fatty acid (FA) profile and expression of key neuronal proteins were analyzed in the cerebral cortex, with intact female mice on standard diet serving as internal controls of brain lipidome composition. Diets containing different concentrations of LC-PUFAs greatly modified total FAs, sphingolipids, and gangliosides in the cerebral cortex. Some of these changes were dependent on ovarian hormones, as they were not detected in ovariectomized animals, and in the case of complex lipids, the effect of ovariectomy was partially or totally reversed by continuous administration of estradiol. However, even though differential dietary LC-PUFA content modified the expression of neuronal proteins such as synapsin and its phosphorylation level, PSD-95, amyloid precursor protein (APP), or glial proteins such as glial fibrillary acidic protein (GFAP), an effect also dependent on the presence of the ovary, chronic estradiol treatment was unable to revert the dietary effects on brain cortex synaptic proteins. These results suggest that, in addition to stable estradiol levels, other ovarian hormones such as progesterone and/or cyclic ovarian secretory activity could play a physiological role in the modulation of dietary LC-PUFAs on the cerebral cortex, which may have clinical implications for post-menopausal women on diets enriched with different proportions of n−3 and n−6 LC-PUFAs.

Impact of the n-6:n-3 long-chain PUFA ratio during pregnancy and lactation on offspring neurodevelopment: 5-year follow-up of a randomized controlled trial.

Evidence regarding the effect of n-3 long-chain polyunsaturated fatty acid (LCPUFA) supplementation during pregnancy on offspring's neurodevelopment is not conclusive. In this analysis, the effect of a reduced n-6:n-3 LCPUFA ratio in the diet of pregnant/lactating women (1.2 g n-3 LCPUFA together with an arachidonic acid (AA)-balanced diet between 15th wk of gestation-4 months postpartum vs control diet) on child neurodevelopment at 4 and 5 years of age was assessed. A child development inventory (CDI) questionnaire and a hand movement test measuring mirror movements (MMs) were applied and the association with cord blood LCPUFA concentrations examined. CDI questionnaire data, which categorizes children as 'normal', 'borderline' or 'delayed' in different areas of development, showed no significant evidence between study groups at 4 (n=119) and 5 years (n=130) except for the area 'letters' at 5 years of age (P=0.043). Similarly, the results did not strongly support the hypothesis that the intervention has a beneficial effect on MMs (for example, at 5 years: dominant hand, fast: adjusted mean difference, -0.08 (-0.43, 0.26); P=0.631). Children exposed to higher cord blood concentrations of docosahexaenoic acid, eicosapentaenoic acid and AA, as well as a lower ratio of n-6:n-3 fatty acids appeared to show beneficial effects on MMs, but these results were largely not statistically significant. Our results do not show clear benefits or harms of a change in the n-6:n-3 LCPUFA ratio during pregnancy on offspring's neurodevelopment at preschool age. Findings on cord blood LCPUFAs point to a potential influence on offspring development.

Reduction of the n–6:n–3 long-chain PUFA ratio during pregnancy and lactation on offspring body composition: follow-up results from a randomized controlled trial up to 5 y of age1–3

It has been hypothesized that the n-6:n-3 (ω-6:ω-3) long-chain polyunsaturated fatty acid (LCPUFA) ratio in the maternal diet during the prenatal and early postnatal phase positively affects the body composition of the offspring. However, only limited data from prospective human intervention studies with long-term follow-up are available. We assessed the long-term effects of a reduced n-6:n-3 LCPUFA ratio in the diets of pregnant and lactating women [1020 mg docosahexaenoic acid (DHA) plus 180 mg eicosapentaenoic acid (EPA)/d together with an arachidonic acid-balanced diet compared with a control diet] on the body weights and compositions of their offspring from 2 to 5 y of age with a focus on the 5-y results. Participants in the randomized controlled trial received follow-up assessments with annual body-composition measurements including skinfold thickness (SFT) measurements (primary outcome), a sonographic assessment of abdominal subcutaneous and preperitoneal fat, and child growth. In addition, abdominal MRI was performed in a subgroup of 5-y-old children. For the statistical analysis, mixed models for repeated measures (MMRMs) were fit with the use of data from each visit since birth (except for MRI). Maternal LCPUFA supplementation did not significantly influence the children's sum of 4 SFTs [means ± SDs at 5 y of age: intervention, 23.9 ± 4.7 mm (n = 57); control, 24.5 ± 5.0 mm (n = 55); adjusted mean difference, -0.5 (95% CI: -2.2, 1.2)], growth, or ultrasonography measures at any time point in the adjusted MMRM model (all P values < 0.05). Results were consistent with abdominal MRI measurements (n = 44) at 5 y of age, which showed no significant differences in subcutaneous and visceral adipose tissue volumes and ratios. The current study provides no evidence that a dietary reduction of the n-6:n-3 LCPUFA ratio in the maternal diet during pregnancy and lactation is a useful early preventive strategy against obesity at preschool age. This trial was registered at clinicaltrials.gov as NCT00362089.

Evaluation of less invasive methods to assess fatty acids from phospholipid fraction: cheek cell and capillary blood sampling

Plasma is the most commonly employed matrix for analyzing fatty acids (FAs), but its extraction is not well accepted in the infant population. The objectives of this study were to evaluate cheek cells and capillary blood as alternatives to plasma sampling for FA analysis and to standardize the methodology. Samples were obtained from 20 children who underwent lipid extraction, phospholipid isolation by Solid Phase Extraction (SPE) in a 96-well plate, methylation, and analysis by fast gas chromatography (GC). A positive correlation was found for most of the FAs, especially long-chain polyunsaturated fatty acids (LC-PUFAs), in cheek cells and capillary blood versus plasma samples (r = 0.32–0.99). No differences were found in the levels of n-6: n-3 PUFA and n-6: n-3 LC-PUFA ratios between cheek cells and capillary blood. These two proposed samples can therefore be used as alternatives to plasma sampling for phospholipid FA analysis, especially LC-PUFAs.

In vivo metabolism of unsaturated fatty acids in Sepia officinalis hatchlings

The transition of Sepia officinalis culture to industrial large scale has been hampered due to bottlenecks related to the limited knowledge on nutritional physiology of the species. Determination of the endogenous ability of S. officinalis hatchlings to metabolise unsaturated fatty acids (FA) may provide new insight on the capability of hatchlings to biosynthesise different FA, as well as lipid classes containing essential fatty acids (EFA). In the present study, cuttlefish hatchlings were incubated with [1-14C]FA including C18 FA (18:1n-9, 18:2n-6, 18:3n-3) and long-chain polyunsaturated fatty acids (LC-PUFA) (20:4n-6 (ARA), 20:5n-3 (EPA) or 22:6n-3 (DHA)), which were added individually as potassium salts bound to bovine serum albumin. As a result, it was possible to investigate the in vivo FA metabolism of S. officinalis hatchlings by following the incorporation of specific [1-14C]FA, which points to the suitability of this methodology to study lipid metabolism of newly hatched cephalopods. The majority of radioactivity incorporated was recovered esterified into polar lipids (PL). A pattern was detected, where [1-14C]DHA, [1-14C]C18 FA and their metabolic products were preferentially esterified into phosphatidylcholine, whereas [1-14C]ARA and [1-14C]EPA were mainly esterified into phosphatidylethanolamine. [1-14C]C18 FA were the most transformed FA with several metabolites produced by elongation and possible desaturation being obtained. As a contrary the radioactivity incorporated into hatchling total lipid (TL) from supplemented [1-14C]LC-PUFA only one elongation product was recovered from the three substrates, except for [1-14C]ARA, where an unidentified product was also detected. The present in vivo results indicated that S. officinalis hatchlings may have capability for the first steps in the biosynthesis of ARA and EPA from 18:2n-6 and 18:3n-3, respectively, including the existence of a desaturase potentially involved. Nonetheless, considering the low desaturation rates detected, this process may not be sufficient to cover EFA demands during development of this species. Therefore, dietary ARA and EPA, as well as DHA, should be supplied during the hatchling stage of Sepia. While designing an inert diet, which ensures normal growth and development of this species during the hatchling stage, the C18 FA and LC-PUFA levels and ratios should be considered, since the esterification pattern detected in the present study suggested competition between these FA for esterification into specific lipid classes. Moreover, considering the observed esterification pattern of LC-PUFA into different lipid classes, it is likely that the DHA/EPA/ARA ratio, rather than DHA/EPA or EPA/ARA ratios, would be of great importance for S. officinalis hatchling development. Statement of relevanceThe transition of S. officinalis culture to industrial large scale has been hampered due to bottlenecks related to the knowledge on nutritional physiology of the species. Due to that fact, using a similar approach to that recently employed to determine the lipid requirements of Octopus vulgaris, the present in vivo study tries to understand the lipid metabolism of unsaturated fatty acids in S. officinalis hatchlings. Information that may assist the design of an accurate inert diet that would ensure normal growth and development of this species during the hatchling stage.The present in vivo results indicated that S. officinalis hatchlings may have capability for the first steps in the biosynthesis of 20:4n-6 (ARA) and 20:5n-3 (EPA) from 18:2n-6 and 18:3n-3, respectively, including the existence of a desaturase potentially involved. Nonetheless, considering the low desaturation rates detected, this process may not be sufficient to cover EFA demands during development of this species. Therefore, dietary ARA and EPA, as well as DHA, should be supplied during the hatchling stage of Sepia.While designing an inert diet, C18 FA and LC-PUFA levels and ratios should also be considered, since the esterification pattern detected in the present study suggested a competition between these FA for esterification into specific lipid classes. Moreover, considering the observed esterification pattern of LC-PUFA into different lipid classes, it is likely that the DHA/EPA/ARA ratio, rather than DHA/EPA or EPA/ARA ratios, would be of great importance for S. officinalis hatchling development.

The effect of long-chain polyunsaturated fatty acids intake during pregnancy on adiposity of healthy full-term offspring at birth.

The adjusted effect of long-chain polyunsaturated fatty acid (LCPUFA) intake during pregnancy on adiposity at birth of healthy full-term appropriate-for-gestational age neonates was evaluated. In a cross-sectional convenience sample of 100 mother and infant dyads, LCPUFA intake during pregnancy was assessed by food frequency questionnaire with nutrient intake calculated using Food Processor Plus. Linear regression models for neonatal body composition measurements, assessed by air displacement plethysmography and anthropometry, were adjusted for maternal LCPUFA intakes, energy and macronutrient intakes, prepregnancy body mass index and gestational weight gain. Positive associations between maternal docosahexaenoic acid intake and ponderal index in male offspring (β=0.165; 95% confidence interval (CI): 0.031-0.299; P=0.017), and between n-6:n-3 LCPUFA ratio intake and fat mass (β=0.021; 95% CI: 0.002-0.041; P=0.034) and percentage of fat mass (β=0.636; 95% CI: 0.125-1.147; P=0.016) in female offspring were found. Using a reliable validated method to assess body composition, adjusted positive associations between maternal docosahexaenoic acid intake and birth size in male offspring and between n-6:n-3 LCPUFA ratio intake and adiposity in female offspring were found, suggesting that maternal LCPUFA intake strongly influences fetal body composition.

Age-dependent effects of cord blood long-chain PUFA composition on BMI during the first 10 years of life

In the present study, we investigated whether n-6 and n-3 long-chain PUFA (LC-PUFA) concentrations in cord blood (CB) serum are associated with BMI up to 10 years of age, after accounting for LC-PUFA composition at 2, 6 and 10 years. The study was based on 388 participants of the German LISAplus (Influence of Lifestyle-Related Factors on the Immune System and the Development of Allergies in Childhood Plus the Influence of Traffic Emissions and Genetics) birth cohort study carried out in Munich. BMI was measured at 2, 6 and 10 years of age. Serum phospholipid fatty acid concentrations were measured by GC in CB and in blood collected at 2, 6 and 10 years of age. The association between n-3 LC-PUFA and n-6 LC-PUFA concentrations and n-6:n-3 LC-PUFA ratio in CB serum glycerophospholipids and BMI z-scores was assessed using linear mixed models adjusted for LC-PUFA composition at follow-up and potential confounders. Interaction terms between time of follow-up and LC-PUFA concentrations in CB were included. There was no consistent association between n-6 and n-3 LC-PUFA concentrations in CB and BMI over time. However, there was a significant interaction between n-6:n-3 LC-PUFA ratio in CB and time of follow-up with respect to BMI (P=0·0415): a negative effect at 2 years; no effect at 6 years; a positive effect at 10 years. BMI up to 10 years of age may be influenced by the n-6:n-3 ratio in CB serum glycerophospholipids in a time-varying fashion. The present results thereby highlight the importance of considering age when examining associations between fatty acid concentrations and BMI.

Breast milk fatty acid profile in relation to infant growth and body composition: results from the INFAT study

There is some evidence that the n-6/n-3 long-chain polyunsaturated fatty acids (LCPUFAs) ratio in early nutrition, and thus in breast milk, could influence infant body composition. In an open-label randomized controlled trial (RCT), 208 healthy pregnant women were allocated to a dietary intervention (supplementation with 1,200 mg n-3 LCPUFAs per day and instructions to reduce arachidonic acid (AA) intake) from the 15th wk of gestation until 4 mo of lactation or to follow their habitual diet. Breast milk LCPUFAs at 6 wk and 4 mo postpartum were related to infant body composition assessed by skinfold thickness (SFT) measurements and ultrasonography during the first year of life. Dietary intervention significantly reduced breast milk n-6/n-3 LCPUFAs ratio. In the whole sample, early breast milk docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and n-3 LCPUFAs at 6 wk postpartum were positively related to the sum of four SFT measurements at age 1. Breast milk AA and n-6 LCPUFAs at 6 wk postpartum were negatively associated with weight, BMI, and lean body mass (LBM) up to 4 mo postpartum. Breast milk n-3 LCPUFAs appear to stimulate fat mass growth over the first year of life, whereas AA seems to be involved in the regulation of overall growth, especially in the early postpartum period.

Effect of dietary intervention to reduce the n-6/n-3 fatty acid ratio on maternal and fetal fatty acid profile and its relation to offspring growth and body composition at 1 year of age

Evidence is accumulating that the long-chain PUFA (LCPUFA) are associated with offspring growth and body composition. We investigated the relationship between LCPUFAs in red blood cells (RBCs) of pregnant women/breastfeeding mothers and umbilical cord RBCs of their neonates with infant growth and body composition ≤ 1 year of age. In an open-label randomized, controlled trial, 208 healthy pregnant women received a dietary intervention (daily supplementation with 1200 mg n-3 LCPUFAs and dietary counseling to reduce arachidonic acid (AA) intake) from the 15th week of gestation until 4 months of lactation or followed their habitual diet. Fatty acids of plasma phospholipids (PLs) and RBCs from maternal and cord blood were determined and associated with infant body weight, body mass index (BMI), lean body mass and fat mass assessed by skinfold thickness measurements and ultrasonography. Dietary intervention significantly reduced the n-6/n-3 LCPUFA ratio in maternal and cord-blood plasma PLs and RBCs. Maternal RBCs docosahexaenoic acid (DHA), n-3 LCPUFAs and n-6 LCPUFAs at the 32nd week of gestation were positively related to birth weight. Maternal n-3 LCPUFAs, n-6 LCPUFAs and AA were positively associated with birth length. Maternal RBCs AA and n-6 LCPUFAs were significantly negatively related to BMI and Ponderal Index at 1 year postpartum, but not to fat mass. Maternal DHA, AA, total n-3 LCPUFAs and n-6 LCPUFAs might serve as prenatal growth factors, while n-6 LCPUFAs also seems to regulate postnatal growth. The maternal n-6/n-3 LCPUFA ratio does not appear to have a role in adipose tissue development during early postnatal life.

Salmon Consumption during Pregnancy Alters Fatty Acid Composition and Secretory IgA Concentration in Human Breast Milk1–4

Fish oil supplementation during pregnancy alters breast milk composition, but there is little information about the impact of oily fish consumption. We determined whether increased salmon consumption during pregnancy alters breast milk fatty acid composition and immune factors. Women (n = 123) who rarely ate oily fish were randomly assigned to consume their habitual diet or to consume 2 portions of farmed salmon per week from 20 wk of pregnancy until delivery. The salmon provided 3.45 g long-chain (LC) (n-3) PUFA/wk. Breast milk fatty acid composition and immune factors [soluble CD14, transforming growth factor-β (TGFβ)1, TGFβ2, and secretory IgA] were analyzed at 1, 5, 14, and 28 d postpartum (PP). Breast milk from the salmon group had higher proportions of EPA (80%), docosapentaenoic acid (30%), and DHA (90%) on d 5 PP compared with controls (P < 0.01). The LC (n-6) PUFA:LC (n-3) PUFA ratio was lower for the salmon group on all days of PP sampling (P ≤ 0.004), although individual (n-6) PUFA proportions, including arachidonic acid, did not differ. All breast milk immune factors decreased between d 1 and 28 PP (P < 0.001). Breast milk secretory IgA (sIgA) was lower in the salmon group (d 1–28 PP; P = 0.006). Salmon consumption during pregnancy, at the current recommended intakes, increases the LC (n-3) PUFA concentration of breast milk in early lactation, thus improving the supply of these important fatty acids to the breast-fed neonate. The consequence of the lower breast milk concentration of sIgA in the salmon group is not clear.