Determination of Flavonoids and Phenolic Acids in the Liver of Wistar Rats after a Dietary Enrichment with Corinthian Currant (Vitis vinifera L., var. Apyrena): A Liquid Chromatography-Tandem Mass Spectrometry Study.

Corinthian currants are dried fruits produced from Vitis vinifera L. var. Apyrena grape. This study investigated the distribution of phenolic compounds in male Wistar rat livers following two distinct Corinthian currant long-term dietary intake protocols (3 and 10% w/w). Method optimization, comparing fresh and lyophilized tissues, achieved satisfactory recoveries (>70%) for most analytes. Enzymatic hydrolysis conditions (37 °C, pH 5.0) minimally affected phenolics, but enzyme addition showed diverse effects. Hydrolyzed lyophilized liver tissue from rats consuming Corinthian currants (3 and 10% w/w) exhibited elevated levels of isorhamnetin (20.62 ± 2.27 ng/g tissue and 33.80 ± 1.38 ng/g tissue, respectively), along with similar effects for kaempferol, quercetin, and chrysin after prolonged Corinthian currant intake. This suggests their presence as phase II metabolites in the fasting-state liver. This study is the first to explore phenolic accumulation in rat liver, simulating real conditions of dried fruit consumption, as seen herein with Corinthian currant.

Corinthian Currants Promote the Expression of Paraoxonase-1 and Enhance the Antioxidant Status in Serum and Brain of 5xFAD Mouse Model of Alzheimer's Disease.

Paraoxonase-1 (PON1), a serum antioxidant enzyme, has been implicated in Alzheimer's disease (AD) pathogenesis that involves early oxidative damage. Corinthian currants and their components have been shown to display antioxidant and other neuroprotective effects in AD. We evaluated the effect of a Corinthian currant paste-supplemented diet (CurD), provided to 1-month-old 5xFAD mice for 1, 3, and 6 months, on PON1 activity and levels of oxidation markers in serum and the brain of mice as compared to a control diet (ConD) or glucose/fructose-matched diet (GFD). Administration of CurD for 1 month increased PON1 activity and decreased oxidized lipid levels in serum compared to ConD and GFD. Longer-term administration of CurD did not, however, affect serum PON1 activity and oxidized lipid levels. Furthermore, CurD administered for 1 and 3 months, but not for 6 months, increased PON1 activity and decreased free radical levels in the cortex of mice compared to ConD and GFD. To probe the mechanism for the increased PON1 activity in mice, we studied the effect of Corinthian currant polar phenolic extract on PON1 activity secreted by Huh-7 hepatocytes or HEK293 cells transfected with a PON1-expressing plasmid. Incubation of cells with the extract led to a dose-dependent increase of secreted PON1 activity, which was attributed to increased cellular PON1 expression. Collectively, our findings suggest that phenolics in Corinthian currants can increase the hepatic expression and activity of antioxidant enzyme PON1 and that a Corinthian currant-supplemented diet during the early stages of AD in mice reduces brain oxidative stress.

Corinthian Currants Supplementation Restores Serum Polar Phenolic Compounds, Reduces IL-1beta, and Exerts Beneficial Effects on Gut Microbiota in the Streptozotocin-Induced Type-1 Diabetic Rat.

The present study aimed at investigating the possible benefits of a dietary intervention with Corinthian currants, a rich source of phenolic compounds, on type 1 diabetes (T1D) using the animal model of the streptozotocin-(STZ)-induced diabetic rat. Male Wistar rats were randomly assigned into four groups: control animals, which received a control diet (CD) or a diet supplemented with 10% w/w Corinthian currants (CCD), and diabetic animals, which received a control diet (DCD) or a currant diet (DCCD) for 4 weeks. Plasma biochemical parameters, insulin, polar phenolic compounds, and inflammatory factors were determined. Microbiota populations in tissue and intestinal fluid of the caecum, as well as fecal microbiota populations and short-chain fatty acids (SCFAs), were measured. Fecal microbiota was further analyzed by 16S rRNA sequencing. The results of the study showed that a Corinthian currant-supplemented diet restored serum polar phenolic compounds and decreased interleukin-1b (IL-1b) (p < 0.05) both in control and diabetic animals. Increased caecal lactobacilli counts (p < 0.05) and maintenance of enterococci levels within normal range were observed in the intestinal fluid of the DCCD group (p < 0.05 compared to DCD). Higher acetic acid levels were detected in the feces of diabetic rats that received the currant diet compared to the animals that received the control diet (p < 0.05). Corinthian currant could serve as a beneficial dietary component in the condition of T1D based on the results coming from the animal model of the STZ-induced T1D rat.

Brain polar phenol content, behavioural and neurochemical effects of Corinthian currant in a rotenone rat model of Parkinson's disease.

OBJECTIVE: Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by the loss of nigral dopaminergic neurons, leading to reduced motor control. A contributing factor for the nigrostriatal degeneration is known to be oxidative stress, while antioxidant/anti-inflammatory properties of natural polyphenols have been suggested to show beneficial effects. The present study questioned the potential neuroprotective effects of supplementary diet with Corinthian currant, using a rat rotenone PD model. METHODS: The alterations in motor activity, brain Corinthian currant polar phenols' accumulation, expression patterns of tyrosine hydroxylase (TH), dopamine transporter (DAT) and brain-derived neurotrophic factor (BDNF) in the nigrostriatal dopaminergic system were determined in rotenone-treated, currant-diet rats and matching controls. RESULTS: Rotenone treatment resulted in motor deficits and TH expression decreases in the nigrostriatal pathway, exhibiting PD-like behavioural motor and neurochemical phenotypes. Interestingly, 38 days Corinthian currant consumption resulted in differential accumulation of polar phenols in mesencephalon and striatum and had a significant effect on attenuating motor deficits and dopaminergic cell loss in substantia nigra pars compacta. In addition, it induced up-regulation of BDNF expression in the nigrostriatal dopaminergic system. DISCUSSION: Taken all together, evidence is provided for the potential neuroprotective influences of Corinthian currant consumption, involving the neurotrophic factor BDNF, in rescuing aspects of PD-like phenotype.

Polar Phenol Detection in Plasma and Serum: Insights on Sample Pre-Treatment for LC/MS Analysis and Application on the Serum of Corinthian Currant-Fed Rats.

Analysis of plasma and serum provides valuable information on the amounts of polar phenols' circulating after ingestion. In the present study, protein precipitation (PPT), liquid-liquid extraction (LLE), solid phase extraction (SPE), enzymatic hydrolysis and their combinations were meticulously evaluated for the extraction of a variety of polar phenolic moieties from plasma and serum. The recovery values of the above methods were compared; satisfactory recoveries (>60%) were attained for most analytes. Polar phenol aglycones undergo degradation with enzymatic hydrolysis; however, their extended phase II metabolism makes enzymatic hydrolysis a mandated process for their analysis in such biofluids. Hence, enzymatic hydrolysis followed by LLE was used for the identification of polar phenols in rats' serum, after the long-term oral consumption of Corinthian Currant. Corinthian Currant is a Greek dried vine product rich in bioactive polar phenolics. Flavonoids and phenolic acids, detected as aglycones, ranged from 0.57 ± 0.08 to 181.66 ± 48.95 and 3.45 ± 1.20 to 897.81 ± 173.96 ng/mL, respectively. The majority of polar phenolics were present as phase II metabolites, representing their fasting state in the blood stream. This is the first study evaluating the presence of polar phenolics in the serum of rats following a long-term diet supplemented with Corinthian Currant as a whole food.

Polar phenol detection in rat brain: Development and validation of a versatile UHPLC-MS method and application on the brain tissues of Corinthian currant (Vitis vinifera L.,var. Apyrena) fed rats.

This study aimed to validate a rapid and selective bioanalytical method, using UHPLC-Orbitrap MS, for the determination of brain polar phenolics and to apply it in rats that orally consumed Corinthian currant for 38 days. Corinthian currant, is a dried vine fruit rich in polar phenolics that potentially penetrate the brain. During method optimization fresh and lyophilized tissues were comparatively studied along with different solid-phase extraction cartridges; satisfactory recoveries (>80%) for almost all analytes were attained using fresh tissues and Oasis® HLB cartridges. Brain regional levels in phenol concentrations were then determined; isoquercetin showed higher concentrations in frontal cortex, hippocampus and cerebellum (14.0 ± 5.5, 6.6 ± 2.0, and 2.9 ± 1.3 ng/g tissue, respectively); rutin and gallic acid in cerebellum and isorhamnetin, quercetin and rutin in hippocampus of the Corinthian currant supplemented rat group compared to the control. This is the first study investigating polar phenolics' accumulation in rat brain after Corinthian currant supplementation.

Host-guest inclusion complexes of hydroxytyrosol with cyclodextrins: Development of a potential functional ingredient for food application.

Hydroxytyrosol (HT), a potent phenolic phytochemical, exerts positive health effects due to its antioxidant properties. However, it is highly reactive to oxygen, light, and heat and presents high instability. Alpha- and beta-cyclodextrin (α-CD, ß-CD) have structures that allow them to encapsulate a variety of hydrophobic molecules. The aim of this study was to examine the outcomes of the inclusion of HT into α-CD and ß-CD. Aqueous solutions of HT and either α-CD or ß-CD were prepared and freeze-drying was applied for the encapsulation, in 1:1 and 2:1 molar ratios. The produced solid complexes were studied and characterized using NMR spectroscopy, differential scanning calorimetry (DSC) and attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR). Encapsulation efficiency (EE%), stability, and in vitro release of the encapsulated complexes under simulated digestion conditions were also evaluated. In both DSC thermograms and FTIR spectra of the inclusion complexes, absence of the characteristic peaks of HT and shifts of the CDs peaks were observed, showing an interaction between the molecules. NMR suggested a stronger complex formed between ß-CD and HT. The EE% of ß-CD/HT (1:1 and 2:1) complexes and α-CD/HT (1:1) complex was found to be higher (83%, 76%, 78%, respectively), compared to α-CD/HT (2:1) (51%). Data obtained support the encapsulation of HT in both CDs, revealing a potential interaction between them and an improvement in HT's thermal stability. Regarding the in vitro release study, both CD complexes had similar behavior and a controlled release of HT in the intestinal site was observed. PRACTICAL APPLICATION: The encapsulation of hydroxytyrosol in cyclodextrins resulted in white amorphous food-grade powders with no aroma and taste. Incorporation of these powders in foods could lead to an increase in their antioxidant content and offer an additional nutritional value.

Mechanistic insight into the capacity of natural polar phenolic compounds to abolish Alzheimer's disease-associated pathogenic effects of apoE4 forms.

Polar phenols found in plant foods have been suggested to act protectively against pathogenic processes underlying Alzheimer's disease (AD), such as oxidative stress. The major risk factor for AD is apolipoprotein E4 (apoE4) and apoE4 forms can affect AD-related processes. It was shown previously that the hereditary apoE4 mutant apoE4[L28P], as well as the apoE4 fragment apoE4-165, induce neuronal oxidative stress. The effect of polar phenols on AD-related pathogenic functions of apoE4 forms is largely unexplored. The aim was to examine the effect of Corinthian currant polar phenolic extract and specific polar phenols resveratrol, quercetin, kaempferol and epigallocatechin gallate on AD-related functions of apoE4 forms. The polar phenolic extract and the individual compounds restored the viability of human neuroblastoma SK-N-SH cells in the presence of lipoprotein-associated apoE4[L28P] and prevented changes in cellular redox status. Furthermore, resveratrol, quercetin, kaempferol and epigallocatechin gallate prevented redox status changes induced by Aß42 uptake in SK-N-SH cells treated with lipid-free apoE4[L28P] or apoE4-165. Investigation of the molecular mechanism of action of these polar phenols showed that resveratrol prevented cellular Aß42 uptake via changes in cell membrane fluidity. Interestingly, kaempferol prevented cellular Aß42 uptake by apoE4[L28P], but not by apoE4-165, due to a modulating effect on apoE4[L28P] secondary structure and stability. The action of quercetin and epigallocatechin gallate could be attributed to free radical-scavenging or other protective activity. Overall, it is shown for the first time that natural compounds could modify the structure of apoE4 forms and ameliorate AD-related pathogenic effects of apoE4 forms.

Corinthian currants finishing side-stream: Chemical characterization, volatilome, and valorisation through wine and baker's yeast production-technoeconomic evaluation.

The industrial currants finishing generates a considerable amount of side-stream (FSS) with great potential for biotechnological exploitation. The chemical composition of FSS generated from the premium quality Vostitsa currants was studied. Its use for wine making (at low temperatures, using both free and immobilized yeast) combined with baker's yeast production (with minor nutrient supplementation), is also proposed. Analysis showed that FSS has a rich volatilome (including Maillard reaction/lipid degradation products), increased antioxidant capacity, and total lipid and phenolic contents, compared to the marketable product (currants). However, acidity levels and the presence of specific volatiles (such as acetate esters and higher alcohols) may be indicative of microbial spoilage. The wines made from FSS were methanol free and contained higher levels of terpenes (indicating hydrolysis of bound forms) and fermentation-derived volatiles, compared to FSS. A preliminary technoeconomic analysis for integrated wine/baker's yeast industrial production, showed that the investment is realistic and worthwhile.

Evaluation of anti-platelet activity of grape pomace extracts.

Platelets aggregation plays a crucial role in atherothrombosis. Therefore, the aim of the present study was to examine the anti-platelet activity of winery by-products extracts, to find the most potent one and to be further analyzed in order to be used for food fortification. For this purpose, grape pomace from four red varieties was extracted with four solvents of different polarity. The extracts' phenolic content, antioxidant capacity and their ability to inhibit human platelet aggregation against PAF, ADP, TRAP were determined by Light Transmission Aggregometry. The ethanolic extract was further analyzed concerning its anti-platelet effect and its chemical composition by LC-MS/MS and GC-MS. The ethanolic and Bligh-Dyer water phase extracts showed the highest phenolic compounds/anthocyanin content and the best antioxidant activity. However, the most potent inhibition of platelet aggregation was revealed by ethanol extracts, followed by the Bligh-Dyer lipoid phase extracts. Ethanolic extract, found to contain micro-constituents such as phospho-compounds, phenolic compounds and fatty acids. The most abundant phenolic compounds were catechin, epicatechin and quercetin and the most abundant fatty acids were linoleic acid (C18:2n6), linolenic acid (C18:3n3) and palmitic acid (C16:0). Ethanolic extract was capable of inhibiting platelets aggregation in a wide range of agonist concentrations and it also seems that its action is sustained when platelets from coronary heart disease patient were used. Ethanol extract of winery by-products exerts a potent anti-platelet effect and its valorization could lead to the production of functional foods with cardioprotective properties.

Water-soluble vitamin content of sun-dried Corinthian raisins (Vitis vinifera L., var. Apyrena).

BACKGROUND: Corinthian raisins are dried vine products, representing approximately 3% of the world dried vine fruit production. The majority of Corinthian raisin production is of Greek origin. Studies on the B-group vitamin content of Corinthian raisins produced in Greece as well as on the effect of region and cultivation altitude on the B-group vitamin content are absent in the literature. RESULTS: Corinthian raisin vitamin content was evaluated by reversed-phase high-performance liquid chromatography after acid and enzymatic hydrolysis in terms of raisin subcategory, i.e. regions of cultivation, crop-to-crop variations, and cultivation altitude. Eight vitamers from five different vitamins were identified and quantified in Corinthian raisins. Vitamin B3 (0.77-2.82 g × 10-2 × kg-1 ) was found to predominate, followed by B6 (0.27-0.37 g × 10-2 × kg-1 ), B1 (0.19-0.22 g × 10-2 × kg-1 ), and B2 (0.10-0.15 g × 10-2 × kg-1 ). B9 content was up to 7.1 g × 10-5 × kg-1 . Minor differences were observed among regions of cultivation, cultivation altitude and crop-to-crop variations. CONCLUSION: This study revealed the presence of several water-soluble vitamins in Corinthian raisins that, together with other health-promoting micronutrients present in the product, further reinforce its place as part of a healthy diet. © 2019 Society of Chemical Industry.

Corinthian raisins (Vitis vinifera L., var. Apyrena) antioxidant and sugar content as affected by the drying process: a 3-year study.

BACKGROUND: Corinthian raisins or Corinthian currants are dried grape products that account for approximately 3% of dried vine fruit production worldwide. Greece is the major Corinthian raisin producer. In this study Corinthian raisins were produced by applying two traditional drying methods, i.e. sun and shade drying. The drying process effect on the raisin phytochemical and sugar content was assessed; samples obtained over three consecutive crops were evaluated to prevail over climatic condition variations. RESULTS: Total phenolic, flavanol, and anthocyanin content were higher in Corinthian raisins dried under shade as compared with the sun-dried ones; individual anthocyanidin-3-O-glucosides were also qualitatively and quantitatively higher under shade drying in most cases. Differences were observed in Corinthian raisin phytochemical content among the crop years evaluated. The drying process applied did not discriminatively affect the sugar content of Corinthian raisin. CONCLUSION: Drying under shade was proven superior to sun drying with respect to Corinthian raisin phytochemicals and could be considered as an appealing drying method for the production of dried raisins and perhaps other dried fruits. © 2018 Society of Chemical Industry.

Virgin Olive Oil as Frying Oil.

Frying is one of the oldest cooking procedures and is still among the most popular ones for food preparation. Due to their unique sensory characteristics, fried foods are consumed often and with pleasure. During frying, part of the oil is absorbed by the food, thereby becoming part of our diet; most interestingly, in the Mediterranean area approximately 50% of total fat intake is provided by cooking fats. Olive oil is the key lipid component of the Mediterranean diet, the health-promoting effects of which have been largely attributed to olive oil intake. Olive oil is unique among vegetable oils due to its desirable lipid profile and some of its minor components. Scientific evidence now indicates that during frying olive oil behavior is usually equal or superior to that of refined vegetable oils. Herein, an overview of virgin olive oil performance under frying is given, with special reference to the fate of olive oil microconstituents. The compositional changes of foods fried in olive oil are also reviewed and discussed in detail.

Sun dried Corinthian currant (Vitis Vinifera L., var. Apyrena) simple sugar profile and macronutrient characterization.

Corinthian currant simple sugar profile was determined by high-pressure liquid chromatography coupled with refractive index detection; the content in other macroconstituents was also assessed. Corinthian currants analyzed were obtained over three consecutive crop years, while the effect of cultivation area altitude was also evaluated. Sugar extraction was optimized with respect to solvent, process, and time. Simple sugar chromatographic analysis was validated for linearity (R2>0.999), limits of detection (6.23-186.66mg/L) and quantification (18.89-565.65mg/L), precision (%RSD<2.67), and recovery (97.99-101.99%). Fructose, glucose, sucrose, maltose, and total sugar content was 32±1%, 32.5±0.6%, 0.40±0.04%, 0.72±0.07% and 66±2%, respectively; moisture, ash, fat, and protein content was 13.8±0.5%, 2.6±0.7%, 0.43±0.06%, and 2.2±0.4%, respectively. Relatively small variations in sugar and other macroconstituent content were detected over the consecutive crops. Cultivation area altitude affected Corinthian currant sugar, fat, and protein content, yet with no remarkable differences.

Amelioration of oxidative and inflammatory status in hearts of cholesterol-fed rats supplemented with oils or oil-products with extra virgin olive oil components.

PURPOSE: The contribution of extra virgin olive oil (EVOO) macro- and micro-constituents in heart oxidative and inflammatory status in a hypercholesterolemic rat model was evaluated. Fatty acid profile as well as α-tocopherol, sterol, and squalene content was identified directly in rat hearts to distinguish the effect of individual components or to enlighten the potential synergisms. METHODS: Oils and oil-products with discernible lipid and polar phenolic content were used. Wistar rats were fed a high-cholesterol diet solely, or supplemented with one of the following oils, i.e., EVOO, sunflower oil (SO), and high-oleic sunflower oil (HOSO) or oil-products, i.e., phenolics-deprived EVOO [EVOO(-)], SO enriched with the EVOO phenolics [SO(+)], and HOSO enriched with the EVOO phenolics [HOSO(+)]. Dietary treatment lasted 9 weeks; at the end of the intervention blood and heart samples were collected. RESULTS: High-cholesterol-diet-induced dyslipidemia was shown by increase in serum total cholesterol, low-density lipoprotein cholesterol, and triacylglycerols. Dyslipidemia resulted in increased malondialdehyde (MDA) and tumor necrosis factor-α (TNF-α) levels, while glutathione and interleukin 6 levels remained unaffected in all intervention groups. Augmentation observed in MDA and TNF-α was attenuated in EVOO, SO(+), and HOSO(+) groups. Heart squalene and cholesterol content remained unaffected among all groups studied. Heart α-tocopherol was determined by oil α-tocopherol content. Variations were observed for heart ß-sitosterol, while heterogeneity was reported with respect to heart fatty acid profile in all intervention groups. CONCLUSIONS: Overall, we suggest that the EVOO-polar phenolic compounds decreased MDA and TNF-α in hearts of cholesterol-fed rats.

Serum lipid profile and inflammatory markers in the aorta of cholesterol-fed rats supplemented with extra virgin olive oil, sunflower oils and oil-products.

Extra virgin olive oil (EVOO) major and minor component anti-inflammatory effect on aorta was evaluated; Wistar rats were fed (9 weeks) on either a high-cholesterol diet (HCD) or a HCD supplemented with oils, i.e. EVOO, sunflower oil (SO), high-oleic sunflower oil (HOSO), or oil-products modified to their phenolic content, i.e. phenolics deprived-EVOO [EVOO(-)], SO enriched with the EVOO phenolics [SO(+)], HOSO enriched with the EVOO phenolics [HOSO(+)]. HCD induced dyslipidemia and resulted in higher aorta adhesion molecules levels at euthanasia. Groups receiving EVOO, EVOO(-), HOSO, HOSO(+) presented higher serum TC and LDL-c levels compared to cholesterol-fed rats; attenuation of aorta E-selectin levels was also observed. In EVOO/EVOO(-) groups, aorta vascular endothelial adhesion molecule-1 (VCAM-1) was lower compared to HCD animals. SO/SO(+) diets had no effect on endothelial dysfunction amelioration. Overall, our results suggest that major and/or minor EVOO constituents improve aorta E-selectin and VCAM-1, while serum lipids do not benefit.

Anthocyanins content and antioxidant capacity of Corinthian currants (Vitis vinifera L., var. Apyrena).

Corinthian currants are dried vine products used in the Mediterranean diet. Currant is produced almost exclusively in Southern Greece; three commercial subcategories are recognised, i.e. Vostizza, Gulf, and Provincial. Thirty-five batches of Corinthian currants obtained over two crop years and belonging to the several subcategories, were evaluated in terms of total and individual anthocyanins, total phenolics and antioxidant capacity. Variations were observed among batches, cultivation regions, and subcategories with respect to the studied parameters. Up to five anthocyanidin-3-O-glucosides were identified and quantified in currants for the first time. Overall, Vostizza currants presented the higher total anthocyanins content and the lower total phenolics content and antiradical activity; reducing power showed no differentiation among currants subcategories and crop years studied. Though the contribution of currants to the daily anthocyanin intake is rather low, the combined presence of other phenolics as well, makes this product a source of hydrophilic antioxidants.

Nutritional evaluation and health promoting activities of nuts and seeds cultivated in Greece.

Available data suggest that genetic as well as environmental factors may influence nuts and seeds nutrients content. In this context nuts and seeds cultivated in Greece were studied. Macronutrients content was in agreement with that from other areas. Total phenolics content was in the range of 43.0 ± 2.1-1512.7 ± 60.7 mg GAE/100 g for chestnut and walnut, respectively. Thirteen to 22 individual phenolics were identified in the studied species. Oleanolic acid was in the range of 0.10-9.03 mg/100 g. Pumpkin seeds contained the higher squalene content (71.6 mg/100 g). ß-Sitosterol predominated in all samples except pumpkin seeds. Tocopherols ranged from 8.9 mg/100 g (chestnut) to 29.3 mg/100 g (almond). Nuts and seeds hydrophilic extracts at quantities corresponding to the estimated daily consumption by the Greeks succeeded in inhibiting LDL oxidation in vitro by increasing lag time 1.1-14.1 times. One serving of nuts or seeds may cover a significant fraction of health promoting microconstituents daily intake.

Breast milk fat concentration and fatty acid pattern during the first six months in exclusively breastfeeding Greek women.

PURPOSE: To determine fat and fatty acid (FA) profile of Greek mother's milk during the first 6 months of exclusive breastfeeding and to examine their correlation with dietary and other maternal characteristics. METHODS: Milk samples and dietary records were obtained by mothers at 1st (n = 64), 3rd (n = 39), and 6th (n = 24) month postpartum. Fatty acid methylesters were separated and quantified by gas chromatography (GC/FID) and fat concentration by the creamatocrit method. RESULTS: At the 3 time points, milk fat concentration ranged between 26.3 and 30.2 g/l (p > 0.05). Milk's FA composition was expressed as weight percentage (% wt/wt of all FAs detected with a C6 to C22 chain length). Maternal macronutrient and FA dietary intake, as well as the FAs' profile in maternal milk, remained constant over the 6 months. Saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA) represented approx. 46, 35, and 18 % of all FAs, while ω6 and ω3 PUFA were 17.4 and 0.8 %, respectively. Body weight gain during pregnancy was positively related to breast milk's concentration in SFA (p < 0.01) and negatively to milk's concentration in MUFA (p < 0.01). Age and parity were also independent factors affecting the FA profile in maternal milk. A strong positive effect was found during the first month postpartum, between mother's PUFA intake and the concentration of PUFA, ω3 fatty acids, docosahexaenoic and linoleic acid (LA) in the milk, while MUFA intake was strongly correlated with the concentration of PUFA, ω6 fatty acids, and LA. CONCLUSION: This study is among few in literature to determine FA profile of breast milk in European populations and verified certain dietary factors that influence this profile. Maternal PUFA and MUFA intake were found to be important factors affecting breast milk's FA profile.