Comparative assessment of erythrocyte sphingolipid levels as potential cardiovascular health markers in women from Libya and Serbia: a small-scale study.

Aim: Cardiovascular diseases (CVDs) represent the major cause of morbidity and mortality worldwide including Libya, where they account for 43% of all deaths. Sphingolipids are involved in the pathology of numerous diseases including cardiovascular diseases and are proposed as potential biomarkers of cardiovascular health that could be more effective compared to traditional clinical biomarkers. The aim of this study was to determine the sphingolipid content in the erythrocyte membrane of Libyan migrant and Serbian resident women. In addition, to examine if sphingolipid levels could be used as a novel indicator of cardiovascular risk, we evaluated possible correlations with some well-established biomarkers of cardiovascular health.Materials and Methods: A total of 13 Libyan and 15 Serbian healthy women participated in the study. The high-performance version thin-layer chromatography (HPTLC) using the image analysis tool JustTLC was applied for quantification of erythrocytes' sphingolipids.Results: Lower mean values of erythrocytes' sphingolipids and cholesterol concentrations were found in the group of Libyan emigrants compared to Serbian resident women. Besides, in this group of apparently healthy women (n = 28), the sphingolipid content of erythrocytes was inversely related to the Omega-3 index (r =-0.492, p = 0.008) and directly linked to vitamin D status (r = 0.433, p = 0.021) and membrane cholesterol levels (r = 0.474, p = 0.011).Conclusion: The erythrocytes' sphingolipid levels should be measured/assessed as an additional biomarker of CV health, by applying a simple and routine method. Still, further investigation in a larger population-specific context is warranted.

Food Composition Data and Tools Online and Their Use in Research and Policy: EuroFIR AISBL Contribution in 2022.

Food, nutrition, and health are linked, and detailed knowledge of nutrient compositions and bioactive characteristics is needed to understand these relationships. Additionally, increasingly these data are required by database systems and applications. This communication aims to describe the contribution to databases and nutrition fields as well as the activities of EuroFIR AISBL; this member-based, non-profit association was founded to ensure sustained advocacy for food information in Europe and facilitate improved data quality, storage, and access as well as encouraging wider exploitation of food composition data for both research and commercial purposes. In addition to the description of its role and main objectives, a snapshot of EuroFIR AISBL's activities over the years is also given using a quantitative research literature analysis approach. The focus of this communication is to provide descriptions and updates of EuroFIR's online tools, i.e., FoodEXplorer, eBASIS, and PlantLIBRA, by highlighting the main uses and applications. Integrating food-related infrastructures and databases, following standardized and harmonized approaches, and considering interoperability and metrological principles are significant challenges. Ongoing activities and future plans of EuroFIR AISBL are highlighted, including, for instance, work within the Food Nutrition Security Cloud (FNS-Cloud) to make food, nutrition, and (food) security data more findable, accessible, interoperable, and ultimately reusable.

The Effect of Fish Oil-Based Foods on Lipid and Oxidative Status Parameters in Police Dogs.

The synthesis, degradation, and reconstruction of the cell membrane as a metabolic pathway of phospholipids is a constant and dynamic process. Fatty acids as bioactive lipid components of plasma and erythrocyte phospholipids as structural lipids have biological roles in the integrity of cell membranes. Fatty acids, depending on the chain length, the degree of saturation, and the synthesis pathways, can alleviate inflammation and oxidative stress caused by excessive exercise. Considering that changing food intake or diet can influence fatty acid phospholipid metabolism, our study aimed to determine the potential benefits of fish-based diets in working (police) dogs undergoing intensive training concerning bioactive lipids such as fatty acids, phospholipids of plasma, and erythrocytes. Fatty acid esters' composition of plasma and erythrocyte phospholipids as a bioactive lipids, in addition to markers of oxidative stress and metabolic parameters, were analysed by GC chromatography. The food was well tolerated by all dogs, and the compliance to the diet was high throughout the study. After the treatment with fish-based food, blood glucose, total, and LDL cholesterol levels were significantly reduced, indicating positive biochemical profiles of dogs. Correlations of fatty acid phospholipid compositions between plasma and erythrocytes have shown that both plasma and erythrocytes could represent markers of omega-3 eicosapentaenoic and docosahexaenoic acid intake levels in dogs. Morover, fish-based food supplementation caused a significant reduction in lipid peroxidation markers. The enrichment of dogs' diets with marine fish could improve oxidative status and improve roles and status of bioactive lipids, such as membrane phospholipids and fatty acids, as its components in plasma and erythrocytes in police dogs exposed to intensive exercise.

Evaluation of Dietary Intake and Anthropometric Status in 1-9-Year-Old Children Living in Serbia: National Food Consumption Survey according to the EU Menu Methodology.

The Serbian Food Consumption Survey among 1-9-year-old-children was conceptualized and conducted in compliance with the principles, established protocols, and guidelines of the EU Menu project between 2017 and 2021. Valid data were collected for 576 individuals (290 1-3-year-old toddlers and 276 3-9-year-old children). Regardless of age and gender category, the majority (68.80%) of children had normal weights according to the Body Mass Index-for-age classification system. The median daily energy intake was 1406.71 kcal with no differences between the settlement types. The overall median contributions of carbohydrates, protein, and fat to the total energy intake were 47.54%, 14.06%, and 37.88%, respectively. The proportions of the macronutrient intake deviated from the dietary reference values with compliance to the recommendations being particularly poor for fat and fiber. The consumption of energy-dense food groups such as meat and meat products, fat and oil, sugar, and confections was more pronounced among older children. The survey results provide a valuable insight into the nutritional status and dietary habits of toddlers and children 1-9 years old living in Serbia. They may serve as an evidence platform for public health programs, a valuable asset for decision-makers, and a reliable reference to guide nutritional policies, diet monitoring, and interventions targeting this population group in the future.

EFFICACY OF A LIPID-LOWERING DIET ON KEY FATTY ACID RATIOS AND OMEGA-3 INDEX IN HYPERLIPIDEMIC SUBJECTS.

Polyunsaturated fatty acid (PUFA) dietary intake, status and serum key fatty acid (FA) ratios may aid in cardiovascular disease-related risk assessment. The aim of this study was to investigate the effects of lipid-lowering diet on key FA ratios in serum phospholipids and omega-3 index in erythrocyte phospholipids in moderately hyperlipidemic subjects. The study included 41 subjects, mean age 56±6 years. Nutritional habits were evaluated by food frequency questionnaire. Participants followed lipid lowering diet for 12 weeks. Energy intake of omega-6 and omega-3 FA was changed from 7.6% and 0.6% to 5.7% and 1.2%, respectively. Marked decrease in four FA ratios in serum phospholipids, i.e., omega-6/omega-3, arachidonic acid (AA)/eicosapentaenoic acid (EPA), AA/docosahexaenoic acid (DHA), AA/(EPA+DHA) and omega-3 index (EPA+DHA) was found in study subjects after lipid-lowering diet. Total cholesterol/high-density lipoprotein (HDL), low-density lipoprotein (LDL)/HDL and triacylglycerol/HDL-cholesterol ratios positively correlated with all FA ratios, and negatively correlated with total omega-3 levels in serum phospholipids and omega-3 index in erythrocytes. Total serum omega-3 levels showed strongest association with lipoprotein ratios and positive correlation with homeostatic model assessment (HOMA) index. In conclusion, lipid-lowering diet resulted in decreased serum key FA ratios, increased omega-3 levels, and improved insulin sensitivity that may lead to a lower risk of cardiovascular disease in subjects with moderate hyperlipidemia.

Polyphenol-Rich Aronia melanocarpa Juice Consumption Affects LINE-1 DNA Methylation in Peripheral Blood Leukocytes in Dyslipidemic Women.

Cardiovascular disease (CVD) is associated with alterations in DNA methylation and polyunsaturated fatty acid (PUFA) profile, both modulated by dietary polyphenols. The present parallel, placebo-controlled study (part of the original clinical study registered as NCT02800967 at www.clinicaltrials.gov) aimed to determine the impact of 4-week daily consumption of polyphenol-rich Aronia melanocarpa juice (AMJ) treatment on Long Interspersed Nucleotide Element-1 (LINE-1) methylation in peripheral blood leukocytes and on plasma PUFAs, in subjects (n = 54, age range of 40.2 ± 6.7 years) at moderate CVD risk, including an increased body mass index, central obesity, high normal blood pressure, and/or dyslipidemia. The goal was also to examine whether factors known to affect DNA methylation (folate intake levels, MTHFR C677T gene variant, anthropometric and metabolic parameters) modulated the LINE-1 methylation levels upon the consumption of polyphenol-rich aronia juice. Experimental analysis of LINE-1 methylation was done by MethyLight method. MTHFR C677T genotypes were determined by the polymerase chain reaction-restriction fragment length polymorphism method, and folate intake was assessed by processing the data from the food frequency questionnaire. PUFAs were measured by gas-liquid chromatography, and serum lipid profile was determined by using Roche Diagnostics kits. The statistical analyses were performed using Statistica software package. In the comparison after vs. before the treatment period, in dyslipidemic women (n = 22), we observed significant decreases in LINE-1 methylation levels (97.54 ± 1.50 vs. 98.39 ± 0.86%, respectively; P = 0.01) and arachidonic acid/eicosapentaenoic acid ratio [29.17 ± 15.21 vs. 38.42 (25.96-89.58), respectively; P = 0.02]. The change (after vs. before treatment) in LINE-1 methylation directly correlated with the presence of MTHFR 677T allele, average daily folate intake, and the change in serum low-density lipoprotein cholesterol but inversely correlated with the change in serum triacylglycerols (R = 0.72, R 2 = 0.52, adjusted R 2 = 0.36, P = 0.03). The current results imply potential cardioprotective effects of habitual polyphenol-rich aronia juice consumption achieved through the modifications of DNA methylation pattern and PUFAs in subjects at CVD risk, which should be further confirmed. Hence, the precision nutrition-driven modulations of both DNA methylation and PUFA profile may become targets for new approaches in the prevention of CVD.

Short-Term Consumption of Pomegranate Juice Alleviates Some Metabolic Disturbances in Overweight Patients with Dyslipidemia.

Pomegranate juice (PJ) has potential positive effects in patients with metabolic disturbances due to a high content of polyphenols. The objective of this study was to evaluate effects of a 2-week consumption of dietary doses of PJ on blood pressure, lipid metabolism, and oxidative stress markers in overweight patients with dyslipidemia. Twenty-four patients, 8 males and 16 females, 40-60 years of age, with established overweight and dyslipidemia were randomly assigned into intervention group, who consumed 300 mL of PJ daily for 2 weeks, or control group. After 2 weeks of juice intake, intervention group had significantly lower diastolic blood pressure, low-density lipoprotein cholesterol, aminotransferase, and activity of glutathione peroxidase. Furthermore, patients who consumed PJ had reduced percentage of docosahexaenoic acid (22:6n-3, DHA) in plasma phospholipids and increased estimated activity of stearoyl-CoA desaturase. In erythrocytes, we found a significant increase in the levels of dihomo-γ- linolenic acid (20:3n-6, DGLA) and DHA, as well as in estimated activity of Δ6 desaturase, and a decrease in estimated activity of Δ5 desaturase. These results show that even a short-term consumption of dietary doses of PJ exerts beneficial effects and affects lipid metabolism in overweight patients with dyslipidemia.

Association of meal timing with dietary quality in a Serbian population sample.

BACKGROUND: The world-wide adoption of Western lifestyles and eating patterns is associated with adverse effects on nutrient intakes. Here we evaluated the relationships between timing of meals and diet quality in Serbia, a Balkan country with a traditional eating pattern that includes the largest meal of the day as a late lunch. METHODS: A dietary survey was done in the Republic of Serbia using a nationally-representative sample of 74 children and 260 non-pregnant adults. Nutrient intakes were calculated from two 24-h recalls. A Dietary Quality Score (DQS) enumerated how many European Union (EU) Science Hub recommendations were met for fruit and vegetables, fiber, saturated fat, sodium, and sugar. We evaluated whether the timing of dietary intakes is associated with DQS and body mass index. RESULTS: The dietary intakes of children ages 10-17 and adults were similar and were high in total fat intake, with an average of 40% of energy from fat. Mean fruit and vegetable intakes of 473 g/day in adults exceeded the minimal EU recommendation. The most worrisome aspects of the Serbian diet were high intakes of saturated fat, sugar and sodium. Lunch was the meal with the highest mean content of energy, followed by breakfast and dinner, and the average time for lunch was 15:15. Consumption of a higher percentage of calories before 16:00 in adults was associated with higher fruit and vegetable intakes and with higher DQS. The subgroup of adults consuming their largest meal after 20:00 had a lower mean age, more men, and a larger percentage was employed outside of the home. There were no associations of meal timing with BMI, but the prevalence of obesity in this population sample was only 13%. CONCLUSIONS: These results indicate that an earlier meal pattern, and especially consuming the largest meal of the day earlier in the day, was associated with better quality diets. Public health efforts are needed to preserve nutrient intakes as the population shifts away from the traditional Serbian eating pattern. Long-term, deterioration of nutrient intakes could contribute to the increasing rates of obesity that have been observed in Serbia and world-wide.

The Effects of Aronia melanocarpa Juice Consumption on the mRNA Expression Profile in Peripheral Blood Mononuclear Cells in Subjects at Cardiovascular Risk.

Foods and food products that contain polyphenols are proposed to modulate risk of cardiovascular disease. The aim of this three-arm, crossover, randomized, double-blind, placebo-controlled intervention study was to examine the impact of Aronia melanocarpa juice (AMJ), high-polyphenol (AMJ treatment, 1.17 g/100 mL polyphenols) and low-polyphenol (dAMJ treatment, 0.29 g/100 mL polyphenols) dose, on the transcriptome in peripheral blood mononuclear cells (PBMC) of 19 subjects at cardiovascular risk. Transcriptome data were obtained by microarray. Bioinformatic functional annotation analysis was performed on both the whole transcriptome datasets and the differentially expressed genes (DEGs). Expression of selected DEGs was validated by RT-qPCR. Administration of AMJ and dAMJ treatments during the two consecutive four-week treatment periods had additive effects on PBMC transcriptome profiles, with the most pronounced and specific effect noticed for AMJ in the last treatment period (TP3) of the trial. Between the high-dose and low-dose treatments in TP3, there was a multitude of overlapping DEGs and DEG-enriched biological processes and pathways, which primarily included immunomodulation and regulation of cell proliferation/death. Increased expression of TNF, IL1B, IL8, RGS1, OSM, and DUSP2 in TP3 was confirmed by RT-qPCR. The results suggest the immunomodulatory effects of prolonged habitual consumption of polyphenol-rich aronia juice in individuals at cardiovascular risk.

Mitochondrial Function and Dysfunction in Dilated Cardiomyopathy.

Cardiac tissue requires a persistent production of energy in order to exert its pumping function. Therefore, the maintenance of this function relies on mitochondria that represent the "powerhouse" of all cardiac activities. Mitochondria being one of the key players for the proper functioning of the mammalian heart suggests continual regulation and organization. Mitochondria adapt to cellular energy demands via fusion-fission events and, as a proof-reading ability, undergo mitophagy in cases of abnormalities. Ca2+ fluxes play a pivotal role in regulating all mitochondrial functions, including ATP production, metabolism, oxidative stress balance and apoptosis. Communication between mitochondria and others organelles, especially the sarcoplasmic reticulum is required for optimal function. Consequently, abnormal mitochondrial activity results in decreased energy production leading to pathological conditions. In this review, we will describe how mitochondrial function or dysfunction impacts cardiac activities and the development of dilated cardiomyopathy.

Investigation of Variations in the Human Urine Metabolome amongst European Populations: An Exploratory Search for Biomarkers of People at Risk-of-Poverty.

SCOPE: According to Eurostat 2016, approximately 119 million European citizens live at-risk-of-poverty (ROP). This subpopulation is highly diverse by ethnicity, age, and culture in the different EU states, but they all have in common a low income that could represent an increased risk of nutrient deficiencies due to poor nutritional habits. This study aims to investigate the human urine metabolome in the search of common biomarkers representing dietary deficiencies amongst European populations at ROP. METHODS AND RESULTS: 2732 urine samples were collected from 1391 subjects across five different European countries, including the United Kingdom, Finland, Italy, Lithuania, and Serbia, and analyzed using 1 H-NMR spectroscopy. The resulting urine metabolome data were explored according to study design factors including economic status, country, and gender. CONCLUSION: Partitioning of the effects derived from the study design factors using ANOVA-simultaneous component analysis (ASCA) revealed that country and gender effects were responsible for most of the systematic variation. The effect of economic status was, as expected, much weaker than country and gender, but more pronounced in Lithuania than in other countries. Citrate and hippurate were among the most powerful ROP biomarkers. The possible relationship between these markers and nutritional deficiencies amongst the ROP population is discussed.

The Development and Validation of Food Atlas for Portion Size Estimation in the Balkan Region.

Assessment of portion sizes is an important factor for the accuracy of food consumption surveys. The objective of this study was to develop and validate a food atlas of commonly consumed foods in the Balkan region in order to improve the accuracy of portion size estimation for food consumption surveys. A list of 135 foods and their portion sizes was based on previously conducted food consumption surveys in this region. Food was cooked, measured and served in three or four portion sizes right before being photographed. A validation study was conducted through the visual perception method. Without receiving training on usage of the food picture book, participants were asked to evaluate two portion sizes of 20 selected foods by comparison with a photo series of each food. Portion sizes were evaluated by 18 nutrition professionals and 17 lay individuals who had no nutritional education. Mean differences and the standard deviations of the mean differences (SD) between the portions estimated by each participant and the served portion were calculated. The percentages of participants who selected the correct, adjacent or distant portion size also were calculated. The number of food items that were quantified within the predefined acceptable range (i.e., mean difference < ∣ 0·7 ∣ and SD < 1) was 16 (80%) among lay individuals and 17 (85%) among nutritional professionals. Among 16 photo series that were assessed as "acceptable," the percentage of all participants, who selected the correct picture, was between 44.3 and 82.9%, with an average of 60.2%. Only three foods were assessed correctly by <50% participants. The percentage of participants who selected the correct or adjacent serving size was above 98% for both lay and professional evaluators. This is the first food atlas containing representative foods and recipes commonly consumed in the Balkan region. However, further adjustments of the methodology should include larger number of food items to be tested, involvement of more participants and provision of training for the users of the food atlas. This food atlas could be used in food consumption surveys in the Balkan region after further testing and validation.

Short-Term Fish Oil Treatment Changes the Composition of Phospholipids While Not Affecting the Expression of Mfsd2a Omega-3 Transporter in the Brain and Liver of the 5xFAD Mouse Model of Alzheimer's Disease.

Long-term fish oil (FO) supplementation is able to improve Alzheimer's disease (AD) pathology. We aimed to determine the impact of short-term fish oil (FO) intake on phospholipids composition and plaque pathology in 5xFAD mice, a widely used animal model of AD. A 3-week-long FO supplementation administered at 3 months of age decreased the number of dense core plaques in the 5xFAD cortex and changed phospholipids in the livers and brains of wild-type (Wt) and 5xFAD mice. Livers of both genotypes responded by increase of n-3 and reciprocal decrease of n-6 fatty acids. In Wt brains, FO supplementation induced elevation of n-3 fatty acids and subsequent enhancement of n-6/n-3 ratio. However, in 5xFAD brains the improved n-6/n-3 ratio was mainly due to FO-induced decrease in arachidonic and adrenic n-6 fatty acids. Also, brain and liver abundance of n-3 fatty acids were strongly correlated in Wts, oppositely to 5xFADs where significant brain-liver correlation exists only for n-6 fatty acids. Expression of omega-3 transporter Mfs2a remained unchanged after FO supplementation. We have demonstrated that even a short-term FO intake improves the phospholipid composition and has a significant effect on plaque burden in 5xFAD brains when applied in early stages of AD pathology.

Anthocyanins and their gut metabolites attenuate monocyte adhesion and transendothelial migration through nutrigenomic mechanisms regulating endothelial cell permeability.

Cardioprotective effects of dietary anthocyanins are partly attributed to their ability to maintain endothelial function. However, the underlying cellular and molecular mechanisms of action are not fully understood. This study aimed to evaluate the effect of anthocyanins and their gut metabolites, at physiologically-relevant conditions, on endothelial cell (EC) function and decipher the underlying molecular mechanisms of action using integrated omics approaches. Primary EC were treated with a mixture of 0.1 µM cyanidin-3-arabinoside, 0.1 µM cyanidin-3-galactoside, 0.1 µM cyanidin-3-glucoside, 0.1 µM delphinidin-3-glucoside, 0.1 µM peonidin-3-glucoside and 0.5 µM 4-hydroxybenzaldehyde for 3 h or a mixture of gut metabolites: 0.2 µM protocatechuic, 2 µM vanillic, 1 µM ferulic and 2 µM hippuric acids for 18 h. Also, successive exposure of EC to both mixtures was performed to mimic anthocyanin pharmacokinetics following their intake. Inflammatory stress was induced using TNFα and monocytes added to assess adhesion and transmigration. Effects of these mixtures on gene, miRNA expression and their potential interaction with cell signalling were investigated. Anthocyanins and their gut metabolites significantly reduced monocyte adhesion and transendothelial migration. Gene expression analysis, using macroarrays, showed that tested compounds modulated the expression of genes involved in cell-cell adhesion, cytoskeleton organisation or focal adhesion. Bioinformatics analyses of gene expression data identified potential transcription factors involved in the observed nutrigenomic effects and signalling proteins regulating their activity. Molecular docking revealed cell signalling proteins to which these bioactives may bind to and potentially affect their activity and the activation of downstream signalling, effects that were in agreement with the results of Western blot analyses. Microarray analysis showed that anthocyanins and their gut metabolites affected miRNA expression in EC, especially those involved in regulation of EC permeability, contributing to the observed changes in EC function. Integration of these results revealed endothelial-protective properties of anthocyanins and their gut metabolites and deciphered new underlying multi-target and multi-layered mode of action.

Assessment of vitamin D intake among Libyan women - adaptation and validation of specific food frequency questionnaire.

Vitamin D deficiency (VDD) has pandemic proportions worldwide. Numerous studies report on high prevalence of VDD in sunny regions like Near East and North Africa (NENA). Previous studies indicated that Libyan population was at risk of VDD. To contribute to the body of evidence, measurement of vitamin D status on children, adults, in Misurata region was conducted, and confirmed with validated dietary intake study. Serum 25(OH)D was analysed using electrochemiluminescence protein binding assay. Existing Food Frequency Questionnaires (FFQ) were adapted to Libyan Women Food Frequency Questionnaire (LW-FFQ). Repeated 24 h dietary recalls and LW-FFQ were employed in vitamin D intake evaluation. LW-FFQ was validated using 24 h dietary recall and vitamin D status as referent methods. The questionnaires included anthropometry and lifestyle information. Vitamin D status assessment revealed inadequate levels (25(OH)D < 50 nmol/l) in almost 80% of participants. Women (25-64 y) were identified as the most vulnerable group with vitamin D inadequacy present in 82% (61.6% had 25(OH)D < 25 nmol/l, and 20.2% had 25-50 nmol/l 25(OH)D). Average Vitamin D intake within the study sample (n = 316) was 3.9 ± 7.9 µg/d, with 92% participants below both Institute of Medicine (IOM) (10 µg/d) and European Food Safety Authority (15 µg/d) recommendations. Measured vitamin D status, in 13% of this group, correlated significantly (p = 0.015) with intake estimates. Based on self-report, consumption of vitamin D supplements does not exist among study participants. Additional lifestyle factors influencing vitamin D status were analysed. Only 2% of study participants spend approximately 11 min on the sun daily, 60.4% were obese, 23.1% were overweight and 71.2% reported low physical activity. These findings confirm previous reports on high prevalence of VDD in women across NENA, and in Libya. The situation calls for multi-sectoral actions and public health initiatives to address dietary and lifestyle habits.

Systematic Review of Observational Studies with Dose-Response Meta-Analysis between Folate Intake and Status Biomarkers in Adults and the Elderly.

BACKGROUND: Dietary reference values for folate intake vary widely across Europe. METHODS: MEDLINE and Embase through November 2016 were searched for data on the association between folate intake and biomarkers (serum/plasma folate, red blood cell [RBC] folate, plasma homocysteine) from observational studies in healthy adults and elderly. The regression coefficient of biomarkers on intake (ß) was extracted from each study, and the overall and stratified pooled ß and SE (ß) were obtained by random effects meta-analysis on a double log scale. These dose-response estimates may be used to derive folate intake reference values. RESULTS: For every doubling in folate intake, the changes in serum/plasma folate, RBC folate and plasma homocysteine were +22, +21, and -16% respectively. The overall pooled regression coefficients were ß = 0.29 (95% CI 0.21-0.37) for serum/plasma folate (26 estimates from 17 studies), ß = 0.28 (95% CI 0.21-0.36) for RBC (13 estimates from 11 studies), and ß = -0.21 (95% CI -0.31 to -0.11) for plasma homocysteine (10 estimates from 6 studies). CONCLUSION: These estimates along with those from randomized controlled trials can be used for underpinning dietary recommendations for folate in adults and elderly.

Effects of anthocyanins and their gut metabolites on adenosine diphosphate-induced platelet activation and their aggregation with monocytes and neutrophils.

Accumulating evidence suggests that anthocyanins play an important role in the cardioprotective effects associated with consumption of anthocyanin-rich foods. These benefits may partly be attributed to their effects on platelets, significant contributors to cardiovascular disease development. This study aimed to investigate the impact of physiologically relevant concentrations of anthocyanins and their metabolites on platelet activation and platelet-leukocyte aggregation. Whole blood from seven healthy volunteers was treated with anthocyanins: cyanidin-3-arabinoside, cyanidin-3-glucoside, cyanidin-3-galactoside, delphinidin-3-glucoside and peonidin-3-glucoside at 0.1 µM concentration or gut metabolites: 4-hydroxybenzaldehyde, protocatechuic, vanillic, ferulic and hippuric acids at 0.5 µM, 0.2 µM, 2 µM, 1 µM, 2 µM concentration, respectively. Markers of adenosine diphosphate-induced platelet activation (P-selectin and GPIIb-IIIa expression) and platelet-monocyte and platelet-neutrophil aggregation were analyzed using flow cytometry. Cyanidin-3-arabinoside, delphinidin-3-glucoside, and peonidin-3-glucoside decreased agonist-induced P-selectin expression, while cyanidin-3-galactoside and cyanidin-3-arabinoside reduced platelet-neutrophil aggregation. Hippuric and protocatechuic acids inhibited P-selectin expression, ferulic acid reduced platelet-monocyte aggregation, while 4-hydroxybenzaldehyde affected P-selectin expression, platelet-neutrophil and monocyte aggregation. Only cyanidin-3-glucoside and protocatechuic acid decreased GPIIb-IIIa expression. These results demonstrate the bioactivity of anthocyanins and their gut metabolites at physiologically relevant concentrations on platelet function and interaction with leukocytes, presenting mechanisms by which they contribute to the beneficial effects of habitual consumption of anthocyanin-rich foods on cardiovascular health.

Effect of urolithins on oxidative stress of colorectal adenocarcinomacells-Caco-2.

Urolithins (UROs) are metabolites derived from ellagic acid (EA) and ellagitannins (ETs) by gut microbiota after consumption of different ETs. The health effects attributed to UROs are numerous and diverse, ranging from antimalarial properties to anticancer activities and regulation of gene expression. The aim of this work was at assessing the effect of URO-A; -B; -C; -D on the oxidative status of colon epithelium using as a model colorectal adenocarcinoma cell line (Caco-2). No significant cytotoxic effects of UROs was noted, with the applied treatments. Supplementation of cell growth medium with a mixture of UROs decreased the level of intracellular reactive oxygen species both after short- and long-term exposure. UROs also affected the activity of antioxidative enzymes within the cell, especially catalase. CONCLUSIONS: At concentrations reached in the lumen of the gut, UROs can exert beneficial effects on the cells by decreasing oxidative stress thus preventing the damage caused by reactive oxygen species.

Validity of the Food Frequency Questionnaire Assessing the Folate Intake in Women of Reproductive Age Living in a Country without Food Fortification: Application of the Method of Triads.

The study aimed to examine the external validity of the Folate Food Frequency Questionnaire (F-FFQ) designed for assessing the folate intake in Serbian women of reproductive age. The F-FFQ was tested against repeated 24 h dietary recalls and correspondent nutritional biomarkers (red blood cells (RBC) and serum folate concentrations) using the method of triads. In a cross sectional study, 503 women aged 18-49 years completed dietary questionnaires and representative validation subsample (n = 50) provided fasting blood samples for biomarker analyses. Correlation coefficients were calculated between each of the dietary methods and three pair-wise correlations were applied for the calculation of validity coefficients. Correlation coefficients observed between F-FFQ and three 24 h recalls were r = 0.56 (p < 0.001) and r = 0.57 (p < 0.001) for total sample and validation group, respectively. Bland-Altman plot and cross-classification analyses indicated good agreement between methods. High validity coefficients were determined between the true intake (I) and dietary assessment methods, F-FFQ (Q) and 24 h dietary recalls (R) (ρQIrbc = 0.871 and ρQIser = 0.814; ρRIrbc = 0.652 and ρRIser = 0.698), and moderate ones for biomarkers (B) (ρBIrbc = 0.428 and ρBIser = 0.421). The F-FFQ is valid instrument for the assessment of dietary folate intake in women living in Serbia, a country without mandatory folic acid food fortification.

Preparatory training attenuates drastic response of the insulin-like growth factor binding protein 1 at the point of maximal oxygen consumption in handball players.

BACKGROUND: Intensive exercise changes physiological need for glucose and several biochemical pathways responsible for its metabolism response. Among them are those which involve insulin, insulin-like growth factor (IGF-1), and IGF-binding proteins (IGFBPs). Different types and degrees of exercise, as well as an athlete's fitness, may induce a range of responses regarding concentrations and time needed for the alteration. The idea of the work was to find out whether and how insulin/IGF axis responds to additional physical activity in the already trained subjects and if so, is the adaptation potentially beneficial from the aspect of metabolic control. METHODS: The effect of 4-week intensive training on campus (preparatory training) on the levels of insulin, IGF-1, and IGFBPs during maximal progressive exercise test (MPET) on a treadmill was compared to the results obtained during MPET conducted after a regular training season of a female elite handball team (n = 17, age: 17 ± 1 years, height: 171 ± 8 cm, weight: 65 ± 8 kg, body mass index: 22 ± 1 kg/m2 at the beginning of the study; there were no significant changes at the end). Serum samples were obtained from players immediately before the test (basal), at the end of the test after reaching the point of maximal oxygen consumption (VO2max), and after recovery. RESULTS: The concentration of insulin decreased at VO2max, but remained higher in players after preparatory training (12.2 ± 2.5 mU/L vs. 8.9 ± 4.4 mU/L, p = 0.049). The level of IGFBP-1 decreased in players at VO2max in either case of training, but it remained much higher in tests performed after the preparatory regime than before (p = 0.029). Concentrations of IGF-1, IGFBP-2, -3, and -4 did not change significantly. CONCLUSION: The inverse relation between insulin and IGFBP-1 was lost during MPET, as these 2 molecules changed in the same direction. The results obtained suggest less severe stress-induced depression of insulin and IGFBP-1 after preparatory training. But another metabolic mechanism cannot be excluded, and that is potentially impaired insulin sensitivity resulting in higher level of IGFBP-1.