Background and aimBlood vessels supply oxygen, nutrients and provide gateways for immune surveillance. Since this network nourishes all tissues, vessel abnormalities contribute to many diseases, such as cancer. One of the potential targets for Docosahexaenoic Acid (DHA) in cancer is suppressing angiogenesis, a process of new blood vessel formation within tumors. In addition, aspirin (ASA) has antineoplastic effects that may be mediated, at least in part, by metabolites derived from acetylated COX-2. We aimed at determining the effect of DHA as well as its metabolites in angiogenesis, using in vitro as well as in vivo models.MethodsEndothelial cell (EC) proliferation, motility and capillary-like tube formation were determined by MTT, wound healing, Boyden and Matrigel assays, respectively. In vivo angiogenesis was measured by the Matrigel sponge model in mice. The biosynthesis of proresolving lipid mediators by ECs was determined by LC-MS-MS.Results and conclusionDHA, but not arachidonic acid (AA), at concentrations consistent with those reached in blood after fish oil supplementation, decreased EC migration in a time- and concentration-dependent manner. Pretreatment with ASA modulated cell migration already after 24 h, while both DHA and ASA decreased migration at longer incubation times without affecting viability. 17-hydroxy-DHA was detected upon incubation with DHA, and increased amounts were observed upon combined treatment with DHA and ASA, an increase that was associated to a synergic effect on EC migration. 17(R)-hydroxy-DHA (17R-HDHA), the metabolite resulting from acetylated COX-2 activity of DHA, reduced EC migration in a concentration-dependent manner. DHA in the presence of ASA, as well as 17R-HDHA, also reduced EC tube formation. These results were confirmed in vivo where both 17R-HDHA or its downstream metabolite 17RResolvinD1 were able to decrease microvessels density in a Matrigel sponge model. Overall, we demonstrated that DHA in the presence of ASA-dependent acetylation of COX-2 showed increased antiangiogenic effects, possibly resulting from its conversion to its hydroxylated derivatives.

Background and Aims : Mutations in the LCAT gene cause Familial LCAT Deficiency (FLD,OMIM#245900), a rare metabolic disorder characterized by severe HDL deficiency, hypertriglyceridemia, and an increased unesterified to total cholesterol ratio. Despite the complete lack of LCAT activity, FLD cases have circulating cholesteryl esters (CE), thus representing a tool to analyze the origin of VLDL cholesteryl esters.Methods: Fifty carriers of LCAT deficiency were included (19 carriers of two LCAT mutant alleles and 31 carriers of one mutant LCAT allele); 39 non affected family members acted as controls. Lipoprotein composition and fatty acid distribution was evaluated in whole plasma and in isolated lipoproteins by chromatography.Results: Plasma levels of CE were significantly reduced and highly variable among carriers of two mutant LCAT alleles (4-87mg/dL), and slightly reduced in heterozygous carriers. Plasma CEFA distribution showed an enrichment in saturated and monounsaturated fatty acids, and a depletion in polyunsaturated fatty acids in carriers of two LCAT alleles. In addition, an increased plasma oleate/linoleate ratio was also noted compared to controls (3.87±2.53 vs 0.57±0.08, P=0.002). Plasma triglyceride-fatty acid distribution was remarkably similar between carriers of LCAT deficiency and controls. CEFA profile in VLDL essentially recapitulated that of plasma. No dramatic changes were instead observed in heterozygous carriers. After fat load, chylomicrons of carriers of LCAT mutations showed CE containing mainly saturated fatty acids.Conclusions: The present study, taking advantage of the availability of a large cohort of carriers of LCAT deficiency, shows that in the absence of LCAT activity CE are still present in VLDL, being derived from ACAT2. Background and Aims : Mutations in the LCAT gene cause Familial LCAT Deficiency (FLD,OMIM#245900), a rare metabolic disorder characterized by severe HDL deficiency, hypertriglyceridemia, and an increased unesterified to total cholesterol ratio. Despite the complete lack of LCAT activity, FLD cases have circulating cholesteryl esters (CE), thus representing a tool to analyze the origin of VLDL cholesteryl esters. Methods: Fifty carriers of LCAT deficiency were included (19 carriers of two LCAT mutant alleles and 31 carriers of one mutant LCAT allele); 39 non affected family members acted as controls. Lipoprotein composition and fatty acid distribution was evaluated in whole plasma and in isolated lipoproteins by chromatography. Results: Plasma levels of CE were significantly reduced and highly variable among carriers of two mutant LCAT alleles (4-87mg/dL), and slightly reduced in heterozygous carriers. Plasma CEFA distribution showed an enrichment in saturated and monounsaturated fatty acids, and a depletion in polyunsaturated fatty acids in carriers of two LCAT alleles. In addition, an increased plasma oleate/linoleate ratio was also noted compared to controls (3.87±2.53 vs 0.57±0.08, P=0.002). Plasma triglyceride-fatty acid distribution was remarkably similar between carriers of LCAT deficiency and controls. CEFA profile in VLDL essentially recapitulated that of plasma. No dramatic changes were instead observed in heterozygous carriers. After fat load, chylomicrons of carriers of LCAT mutations showed CE containing mainly saturated fatty acids. Conclusions: The present study, taking advantage of the availability of a large cohort of carriers of LCAT deficiency, shows that in the absence of LCAT activity CE are still present in VLDL, being derived from ACAT2.

exacerbations (PEx).New airway clearance techniques (ACTs) adapted to individual needs are still being sought to achieve the best effect of airway clearance.The primary aim of this study was to evaluate the efficacy of Simeox -a new ACT in subjects with CF who were hospitalized due to PEx.Methods: CF Patients aged 10-18 admitted to hospital and requiring intravenous (IV) antibiotic therapy due to PEx were allocated consecutively (1:1) to Simeox arm or classical CP arm (without Simeox).Patients performed spirometry and multiple breath nitrogen washout (N 2 MBW) for Lung Clearance Index (LCI) assessment on admission and prior to discharge.Results: Fourty eight CF patients were included in the study: 24 treated with Simeox and 24 using usual CP.In both groups the spirometry parameters increased significantly after IV antibiotic therapy.Significant improvement in MEF 25 was observed only in patients treated with Simeox ( p < 0.01).A similar trend was observed towards lowering LCI ratio in both.No side effect was observed for each therapy.Drainage with Simeox was not painful for any of the patients and about 80% of them did not feel any fatigue.All patients felt comfortable and >80% learned quickly the use of new ACT.Conclusion: Spirometry parameters increased significantly in CF patients treated for PEx with IV antibiotic therapy and intensive CP.Simeox may improve the drainage of the central and peripheral airways.This new ACT was safe, well-tolerated method of CP and can be considered an option in treatment of PEx.

This letter was reposted with corrections on September 13, 2018.

Curcumin, a polyphenol extracted from the rhizome of turmeric (Curcuma longa L.), is considered the component responsible for the different biological activities of turmeric. However, given the limited bioavailability of this molecule, different formulations have been prepared. One of these is MERIVA®, wherein curcumin is present in a complex with phosphatidylcholine. To the best of our knowledge, only one study [1] evaluated the peripheral bioavailability of MERIVA®, and a lack of information exists on its distribution within the brain. On these bases, the aim of our study was to evaluate the bioavailability of curcumin in the rat frontal lobe and hippocampus after a sub-chronic treatment with MERIVA®. Specifically, adult male rats received MERIVA® by oral gavage (134 mg/kg/die as curcuminoids equivalent) for 5 days in comparison with a single administration setting and the concentration of curcumin was assessed in the brain tissues at several time points (30 min, 1h, 2h and 3h) by using liquid chromatography coupled with mass spectrometry (LC-MS/MS). These analyses were paralleled by the assessment of curcumin and its metabolite curcumin-glucuronide at plasma levels. After repeated MERIVA® administration, curcumin reached the brain with a specific temporal profile. In particular, in the frontal lobe it was already present after 30 min (9 pg/mg), peaked at 1h (20 pg/mg) and tended towards normalization after 3h (2 pg/mg). Conversely, only very low levels of curcumin were found after acute administration. These results support the use of curcumin in brain diseases and add important information on the bioavailability of one of its formulations.

Background/objectives: Blood polyunsaturated fatty acids (PUFA) are involved in allergy development, but the etiological role of n-6 and n-3 PUFA is still controversial. A European multicenter study of children (IDEFICS) provided the opportunity to explore the cross-sectional association between fatty acids (FA) and allergy. Subjects/methods: Blood FA levels were measured in 2600 children aged 2–9 years and were recorded as the percentage of weight of all FA detected. Logistic regression of allergy status on FA components was adjusted for age, sex, country, body mass index, family history of allergic disease, breast-feeding, and number of siblings. The results were given as odds ratios (OR) for current vs no allergy ever and an increase in FA by 1 s.d. Results: Overall, higher proportions of n-6 PUFA were associated with higher odds of allergy (OR=1.21 (1.05, 1.40)). Monounsaturated FA (MUFA) were associated with reduced risk for allergy (OR=0.75 (0.65, 0.87)), whereas saturated FA did not differ by allergy status. The strongest associations were observed in children <4 years old, with ORs of allergy given as 1.62 (1.15, 2.29) for n-3 PUFA and 0.63 (0.42, 0.95) for MUFA. With regard to individual FA, these associations were independently observed for docosapentaenoic acid (22:5 n-3) and oleic acid (18:1 n-9). Conclusions: Both PUFA subtypes were positively associated with allergy in an age-dependent manner, whereas MUFA was associated with less allergy. The observation of high proportions of n-3 PUFA in allergic children younger than 4 years might help to understand the nature of early onset of atopic disease.

BACKGROUND/OBJECTIVES:Fatty acids are hypothesized to influence cardiovascular disease risk because of their effect on inflammation. The aim of this study is to assess the relationship between whole-blood fatty acids (WBFAs) and high-sensitivity C-reactive protein (hs-CRP) in European children.SUBJECTS/METHODS:A total of 1401 subjects (697 boys and 704 girls) aged between 2 and 9 years from the IDEFICS (Identification and prevention of Dietary- and lifestyle-induced health EFfects in Children and infantS) study were measured in this cross-sectional analysis. The sample was divided into three categories of hs-CRP. Associations between WBFA and hs-CRP were assessed by logistic regression models adjusting for body mass index (BMI), country, age, breastfeeding, mother's education and hours of physical activity.RESULTS:Linoleic acid (LA) (P=0.013, 95% confidence interval (CI): 0.822-0.977) and sum of n-6 WBFA (P=0.029, 95% CI: 0.866-0.992) concentrations were associated with lower concentrations of hs-CRP in boys. In girls, a high ratio of eicosapentaenoic acid (EPA)/arachidonic acid (AA) was associated (P=0.018, 95% CI: 0.892-0.989) with lower hs-CRP concentrations. In contrast, sum of blood n-6 highly unsaturated fatty acids (P=0.012, 95% CI: 1.031-1.284), AA (P=0.007, 95% CI: 1.053-1.395) and AA/LA ratio (P=0.005, 95% CI: 1.102-1.703) were associated (P<0.05) with higher concentrations of hs-CRP in girls.CONCLUSIONS:The n-6 WBFAs (sum of n-6 FA and LA) were associated with lower hs-CRP in boys and with higher hs-CRP in girls (AA, sum of n-6 highly unsaturated and AA/LA ratio). More studies are needed to identify the optimal levels of WBFAs to avoid low-grade inflammation in children considering the differences by sex and BMI.

OBJECTIVES:To establish reference values for fatty acids (FA) especially for n-3 and n-6 long-chain polyunsaturated FAs (LC PUFA) in whole-blood samples from apparently healthy 3-8-year-old European children. The whole-blood FA composition was analysed and the age- and sex-specific distribution of FA was determined.DESIGN AND SUBJECTS:Blood samples for FA analysis were taken from 2661 children of the IDEFICS (identification and prevention of dietary- and lifestyle-induced health effects in children and infants) study cohort. Children with obesity (n=454) and other diseases that are known to alter the FA composition (n=450) were excluded leaving 1653 participants in the reference population.MEASUREMENTS:The FA composition of whole blood was analysed from blood drops by a rapid, validated gas chromatographic method.RESULTS:Pearson correlation coefficients showed an age-dependent increase of C18:2n-6 and a decrease of C18:1n-9 in a subsample of normal weight boys and girls. Other significant correlations with age were weak and only seen either in boys or in girls, whereas most of the FA did not show any age dependence. For age-dependent n-3 and n-6 PUFA as well as for other FA that are correlated with age (16:0, C18:0 and C18:1n-9) percentiles analysed with the general additive model for location scale and shape are presented. A higher median in boys than in girls was observed for C20:3n-6, C20:4n-6 and C22:4n-6.CONCLUSIONS:Given the reported associations between FA status and health-related outcome, the provision of FA reference ranges may be useful for the interpretation of the FA status of children in epidemiological and clinical studies.

Long chain polyunsaturated fatty acids (LC-PUFA), especially the Omega 3, modulate key functions in the body. Their circulating levels are representative of their "status", and may vary at different ages.We have compared the FA status in Italian subjects from neonates to adulthood, assessed through FA analysis of blood drops from fingertips.Data from four cohorts of Italian subjects (total number 1835), have been pooled in four age-groups: neonates (4 days, n=81), children (2-9 years, n=728), adults (40-59 years, n=434) and elderly (60-79 years, n=592).LC-PUFA of both series (Omega 3 and 6) are higher in the blood of neonates than at subsequent ages, reflecting the efficient transfer of these FA from mothers to the fetus. In contrast, the lowest levels of Omega 3 PUFA, especially of DHA, are found in children, probably reflecting inadequate dietary intakes, with possible consequences on the health status at subsequent ages. (C) 2013 Elsevier Ltd. All rights reserved.

Low plasma concentrations of docosahexaenoic acid (DHA) are reported in unsupplemented cystic fibrosis (CF) patients. Forty-one CF patients aged from 6 to 12 years were randomized to receive high-dose DHA (100 mg/kg/day in the first month and 1g per day thereafter through a 12-month supplementation) or placebo (germ oil). Primary outcome was percentage change in plasma AA:DHA ratio. Secondary outcomes were changes in the number of pulmonary exacerbations compared to previous year, lung function, BMI, skinfold thicknesses, and body composition assessed by DXA and in serum concentrations of C-reactive protein, cytokines and vitamin (α-tocopherol and retinol). Compared to the control group plasma AA:DHA ratio decreased in the intervention group after 6 months (median percentage changes: -73% in the intervention group vs. -10% in the control group, P=0.001). No differences were detected between groups for secondary outcomes. Despite a decrease of the AA/DHA ratio, DHA supplementation for one year did not induce any significant biochemical and clinical improvement in CF patients.

Background Biochemical predictors of infants' growth changes are not available. Objectives We tested whether retinol-binding protein (RBP), docosahexaenoic acid and insulin (I) measured within 72?h from birth are associated with growth changes in infants born to mothers with gestational diabetes mellitus (GDM). Methods Fifty-six children, 32 born to diabetic mothers treated with insulin (GDM-I) and 24 born to diabetic mothers treated with diet (GDM-D), were evaluated at 0, 1, 3, 6 and 12 months of life. Results At multivariable regression performed using generalized estimating equations, early RBP levels and maternal body mass index were associated to average weight changes and early RBP and insulin levels to average length changes, respectively. There was no difference between GDM-I and GDM-D infants. Conclusions This exploratory study suggests that early RBP levels may be a predictor of growth changes.

Background and aim: Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in children. We tested whether dietary supplementation with docosahexaenoic acid (DHA) can decrease liver fat content in children with NAFLD.Methods and results: We performed a randomized controlled trial of DHA supplementation (250 mg/day and 500 mg/day) vs. placebo in 60 children with NAFLD (20 children per group). The main outcome was the change in liver fat as detected by ultrasonography after 6, 12, 18 and 24 months of treatment. Secondary outcomes were changes in triglycerides, alanine transaminase (ALT), body mass index (BMI) and homeostasis model assessment of insulin resistance (HOMA). The odds of more severe versus less severe liver steatosis decreased to the same degree at 6 months in children treated with DHA 250 mg/day and DHA 500 mg/day vs. placebo and persisted virtually unmodified for 24 months (OR <= 0.02, p <= 0.05 for all time points). Triglycerides were lower in the DHA groups than in the placebo group at any time point and ALT was lower in these groups from month 12 onwards. HOMA was lower in the DHA 250 mg group vs. placebo at months 6 and 12.Conclusion: DHA supplementation improves liver steatosis in children with NAFLD. Doses of 250 mg/day and 500 mg/day of DHA appear to be equally effective in reducing liver fat content. (C) 2012 Elsevier B. V. All rights reserved.

290 Oral DHA supplementation in children with cystic fibrosis: a randomized placebo-controlled study G. Alicandro1, R. Gagliardini2, B. Santini3, P. Rise’4, A. Biffi1, A.S. Tirelli1, R.M. Tiso1, L. Valmarana1, N. Cirilli2, C. Colombo1. 1Fond. IRCCS Ca’ Granda Osp. Maggiore Policlinico, CF Center, Universita degli studi di Milano, Milan, Italy; 2CF Center, U.O. Medicina Pediatrica, A.O.U. Ospedali Riuniti “G. Salesi”, Ancona, Italy; 3CF Center, Centro Pediatrico Pneumologia Pediatrica Ospedale Regina Margherita, Torino, Italy; 4Universita degli studi di Milano, Dip. Di Scienze Farmacologiche, Milano, Italy

The effects of a n-3 fatty acid (FA) enriched yoghurt on blood FA were investigated in 23 females and 23 males, who consumed 2 yoghurts/day for 6 weeks, providing 64 mg EPA and 74 mg DHA. Blood samples were collected before treatment, at the beginning, end, and after wash out. Yoghurt intakes increased blood EPA (35%) and DHA (11%) with different trends of changes in women versus men. In conclusion, the daily consumption of a yoghurt containing small amounts of n-3 FA significantly increases blood n-3 FA, providing a useful approach to reach the recommended intakes in populations.