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.

Associations of fatty acids composition and estimated desaturase activities in erythrocyte phospholipids with biochemical and clinical indicators of cardiometabolic risk in non-diabetic Serbian women: the role of level of adiposity.

Introduction: Fatty acids (FAs) composition and desaturase activities can be altered in different metabolic conditions, but the adiposity-independent associations with clinical and biochemical indicators of cardiometabolic risk are still unclear. This study aimed to analyze the associations of FAs composition and estimated desaturase activities with anthropometric, clinical, and biochemical cardiometabolic risk indicators in non-diabetic Serbian women, and to investigate if these associations were independent of the level of adiposity and other confounders. Methods: In 76 non-diabetic, otherwise healthy Serbian women, aged 24-68 years, with or without metabolic syndrome or obesity (BMI=23.6±5.6 kg/m2), FA composition in erythrocyte phospholipids was measured by gas-liquid chromatography. Desaturase activities were estimated from product/precursor FAs ratios (D9D:16:1n-7/16:0; D6D:20:3n-6/18:2n-6; D5D:20:4n-6/20:3n-6). Correlations were made with anthropometric, biochemical (serum glucose, triacylglycerols, LDL-C, HDL-C, ALT, AST, and their ratios) and clinical (blood pressure) indicators of cardiometabolic risk. Linear regression models were performed to test the independence of these associations. Results: Estimated desaturase activities and certain FAs were associated with anthropometric, clinical and biochemical indicators of cardiometabolic risk: D9D, D6D, 16:1n-7 and 20:3n-6 were directly associated, while D5D and 18:0 were inversely associated. However, the associations with clinical and biochemical indicators were not independent of the associations with the level of adiposity, since they were lost after controlling for anthropometric indices. After controlling for multiple confounders (age, postmenopausal status, education, smoking, physical activity, dietary macronutrient intakes, use of supplements, alcohol consumption), the level of adiposity was the most significant predictor of desaturase activities and aforementioned FAs levels, and mediated their association with biochemical/clinical indicators. Vice versa, desaturase activities predicted the level of adiposity, but not other components of cardiometabolic risk (if the level of adiposity was accounted). While the associations of anthropometric indices with 16:1n-7, 20:3n-6, 18:0 and D9D and D6D activities were linear, the associations with D5D activity were the inverse U-shaped. The only adiposity-independent association of FAs profiles with the indicators of cardiometabolic risk was a positive association of 20:5n-3 with ALT/AST ratio, which requires further exploration. Discussion: Additional studies are needed to explore the mechanisms of the observed associations.

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.

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.

Aronia juice consumption prior to half-marathon race can acutely affect platelet activation in recreational runners.

Long-distance running, especially in non-professional runners, can increase cardiac arrest risk by enhancing platelet activation and aggregation. Polyphenols can exert cardioprotective effects by positively influencing platelet function. This study aimed to examine the acute effects of polyphenol-rich aronia juice consumption, before simulation of a half-marathon race, on platelet activation and aggregation with leukocytes in recreational runners. In this acute crossover study,10 healthy male runners (age 30.8 ± 2.3 years) consumed breakfast with 200 mL of aronia juice or 200 mL of placebo. They warmed-up and ran a simulated half-marathon race (21.1 km). Blood was collected at baseline, and at 15 min, 1 h, and 24 h after the run. All variables were analyzed with 4 (time) × 2 (group) ANOVA with repeated measures on both factors. Results revealed a significant effect of group on platelet activation parameters: P-selectin and GPIIb-IIIa expressions significantly decreased in the aronia group compared with the placebo group (F[1,9] = 10.282, p = 0.011 and F[1,9] = 7.860, p = 0.021, respectively). The effect of time was significant on both platelet aggregation markers: platelet-monocyte and platelet-neutrophil aggregates were significantly lower after the race (F[3,7] = 4.227, p = 0.014 and F[3,7] = 70.065, p = 0.000, respectively), with changes more pronounced in the later. All effects remained when platelets were exposed to an agonist. These results suggest that aronia consumption could counteract the half-marathon race-induced changes in platelet function. Novelty Aronia juice consumption significantly decreased the expression of platelet activation markers but did not affect platelet aggregation. The race itself did significantly reduce platelet-neutrophil aggregation. Aronia juice may serve as a supplement beverage for recreational runners to alleviate enhanced platelet reactivity caused by prolonged running.

Vitamin D and cardio-metabolic biomarkers: small-scale comparative study between Libyan migrants and resident women in Serbia.

Libyan women are at high risk of vitamin D deficiency, mostly due to their lifestyle and low exposure to sun. In the last decades, Libyan residents have been forced to seek refuge in countries such as Serbia, a country with high incidence of cardio-metabolic diseases. Serbian residents tend to be deficient in vitamin D, mostly due to the lack of vitamin D fortification policy. The aim of this study was to evaluate vitamin D status in Libyan adult women migrating to Serbia, with the assessment of cardio-metabolic and nutritional biomarkers, including erythrocytes fatty acid composition, magnesium concentration, and dietary intake. The same markers were measured in Serbian women, and comparisons between the groups were made. Despite low vitamin D dietary intake in both study groups, we observed lower plasma vitamin D status in Libyan women. This was accompanied by a significantly lower concentration of magnesium in Libyan women. Libyan women had significantly higher omega-3 index and lower n-6/n-3 ratio in erythrocytes' phospholipids. We observed significant negative correlation between vitamin D and n-6 polyunsaturated fatty acids (PUFA) concentrations in both study groups. Despite lower vitamin D status in the Libyan group, erythrocyte fatty acid composition, along with blood lipids' concentrations, indicated a lower cardiovascular risk. Based on our results, the discrepancy in the vitamin D status could not be ascribed to the participants' dietary intake of the micronutrient, rather is potentially associated with ethnic-specific cardio-metabolic profile, which should be confirmed in larger cohorts.

Chokeberry Juice Containing Polyphenols Does Not Affect Cholesterol or Blood Pressure but Modifies the Composition of Plasma Phospholipids Fatty Acids in Individuals at Cardiovascular Risk.

Chokeberry polyphenols have been suggested to reduce cholesterol and blood pressure and thus protect against cardiovascular diseases (CVD), but the evidence in humans is limited and inconsistent. This randomized double-blinded three-parallel groups trial investigated the changes in various anthropometric and clinical biomarkers, and in plasma phospholipids fatty acids (PPFA) in volunteers at cardiovascular risk after a four-week intervention with 100 mL/day of (1) chokeberry juice with a high-dose of polyphenols (1177.11 mg gallic acid equivalents, GAE); (2) chokeberry juice with a low-dose of polyphenols (294.28 mg GAE) and; (3) a nutritionally matched polyphenol-free placebo drink. Our results indicate that the intake of chokeberry juice containing either the low or the high dose of polyphenols cannot be linked with a reduction in total- and low-density lipoprotein (LDL)cholesterol or in systolic (SBP) and diastolic (DBP) blood pressure in comparison with the consumption of the placebo drink. However, we found evidence of moderate changes in the PPFA, i.e., increased saturated fatty acids (SFA), mostly palmitic acid, and reduced n-6 polyunsaturated fatty acids (PUFA), principally linoleic acid (LA) with the intake of chokeberry against the placebo. These effects may be associated with the polyphenols but we could not differentiate a clear dose-response effect. Further research is still needed to elucidate the contribution of the polyphenolic fraction to the potential cardiovascular effects of the chokeberry and to build up the evidence of its potential benefit via the modulation of PPFA composition.

Aronia (Aronia melanocarpa) Polyphenols Modulate the Microbial Community in a Simulator of the Human Intestinal Microbial Ecosystem (SHIME) and Decrease Secretion of Proinflammatory Markers in a Caco-2/endothelial Cell Coculture Model.

SCOPE: To explore the mechanisms behind the health effects of Aronia (Aronia melanocarpa), the microbial community modulating and anti-inflammatory effects of Aronia polyphenols are investigated by combining the similutor of the human intestinal microbial ecosystem (SHIME) with a coculture of intestinal and endothelial cells. RESULTS: Administration of Aronia juice (6.5g L-1 ) to the SHIME for 2 weeks increases the abundance of firmicutes to 92% in the ascending colon (AC), 85% in the transverse colon (TC), and 82% in the descending colon (DC; p < 0.001), proteobacteria (6.7% in AC, p < 0.001), and Akkermansia (14% in TC and 18% in DC, p < 0.001) and decreases the abundance of Bifidobacterium species, associated with a decrease of acetate and increase of propionate and butyrate, whereas no significant difference is observed upon placebo juice treatment. After addition of the digests to TNF-α challenged Caco-2/endothelial cocultures, intercellular adhesion molecule (ICAM)-1, IL-8, and monocyte chemoattractant protein-1 levels are significantly downregulated. Interestingly, Aronia juice treats digests from each colon compartment resulting in a stronger decrease of the ICAM-1 secretion (up to 73%, p < 0.001) compared to their corresponding placebo treated digests, thereby pointing to a polyphenol-dependent effect. CONCLUSIONS: Aronia polyphenols modulate intestinal microbial composition, induce beneficial short chain fatty acid production, and prevent inflammatory stress in endothelial cells. This opens perspectives for the use of Aronia polyphenols as prebiotics in the context of intestinal and cardiovascular health.

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.