In this work we applied a multimodal approach to define the age- and atherosclerosis-related biochemical and functional alterations in red blood cells (RBCs) in ApoE/LDLR^-/- mice. Our results revealed that age-related changes in RBCs, such as decreases in RBC deformability and mean height, were more pronounced in ApoE/LDLR^-/- mice than in age-matched control mice (C57BL/6J). The decreases in phospholipid content and level of lipid unsaturation were accompanied by an increase in cholesterol esters and esterified lipids in RBC membranes in aged C57BL/6J mice. The age-related decrease in the phospholipid content was more pronounced in ApoE/LDLR^-/- mice. In contrast, the increase in the total lipid content in RBC membranes occurred only in ApoE/LDLR^-/- mice with advanced atherosclerosis. The age-related alterations also included a decrease in the ratio of turns to α-helices in the secondary structure of hemoglobin (Hb) inside intact RBCs. On the other hand, an increase in the ratio of unordered conformations to α-helices of Hb was observed only in ApoE/LDLR^-/- mice and occurred already at the age of 5-weeks. This was related to hypercholesterolemia and resulted in an increased oxygen-carrying capacity. In conclusion, progressive mechanical and functional alterations of RBCs in aged ApoE/LDLR^-/- mice were more pronounced than in age-matched C57BL/6J mice. Although, several biochemical changes in RBCs in aged ApoE/LDLR^-/- mice recapitulated age-dependent changes observed in control mice, some biochemical features of RBC membranes attributed to hypercholesterolemia were distinct and could contribute to the accelerated deterioration of RBC function in ApoE/LDLR^-/- mice.

The aim of this study was to evaluate the anti-atherosclerotic effect of pomegranate seed oil as a source of conjugated linolenic acid (CLnA) (cis-9,trans-11,cis-13; punicic acid) compared to linolenic acid (LnA) and conjugated linoleic acid (CLA) (cis-9,trans-11) in apoE/LDLR^−/− mice. In the LONG experiment, 10-week old mice were fed for the 18 weeks. In the SHORT experiment, 18-week old mice were fed for the 10 weeks. Diets were supplied with seed oils equivalent to an amount of 0.5% of studied fatty acids. In the SHORT experiment, plasma TCh and LDL+VLDL cholesterol levels were significantly decreased in animals fed CLnA and CLA compared to the Control. The expression of PPARα in liver was four-fold increased in CLnA group in the SHORT experiment, and as a consequence the expression of its target gene ACO was three-fold increased, whereas the liver’s expression of SREBP-1 and FAS were decreased in CLnA mice only in the LONG experiment. Punicic acid and CLA isomers were determined in the adipose tissue and liver in animals receiving pomegranate seed oil. In both experiments, there were no effects on the area of atherosclerotic plaque in aortic roots. However, in the SHORT experiment, the area of atherosclerosis in the entire aorta in the CLA group compared to CLnA and LnA was significantly decreased. In conclusion, CLnA improved the lipid profile and affected the lipid metabolism gene expression, but did not have the impact on the development of atherosclerotic plaque in apoE/LDLR^−/− mice.

The objective of this study was to produce eggs enriched with conjugated linoleic acid (CLA) and ameliorate their fatty acid profile using the appropriate combination of dietary CLA with or without vegetable oils (olive oil or rapeseed oil) and vitamin E. In Experiment 1, 25-week-old laying hens were randomly distributed into eight groups of nine. Birds were fed with a standard diet with four different levels of CLA (0.0, 0.5, 0.75, 1.0%) and vegetable oils (olive oil or rapeseed oil, both in the amount of 1.46%). In Experiment 2, hens were randomly distributed into 12 groups of nine. The same four levels of CLA with three doses of vitamin E (0; 150; 300 mg/kg of diet) were applied. In both experiments, eggs were collected twice (at 4 and 8 weeks) for fatty acid profiling using GCMS. The differences between treatment means were considered significant at P<0.05. CLA treatments significantly increased the content of CLA, saturated fatty acids (SFA), and significantly decreased the content of monounsaturated fatty acids (MUFA) in the egg yolk, whereas levels of polyunsaturated fatty acids (PUFA) were unaffected. The vegetable oils used did not prevent the negative effects of CLA effectively. Only after eight weeks of experiment 1 SFA levels were significantly lower, but MUFA levels were significantly higher in groups fed with rapeseed oil compared to groups fed with olive oil. In experiment 2, the addition of vitamin E to the hen diet did not have an essential influence on the lipid profile of egg yolks.

The aim of this study was to investigate and confirm the properties of eggs produced by laying hens fed a diet consisting of pomegranate seed oil as a source of CLnA and flaxseed oil as a source of α-linolenic acid. The study involved determining the chemical composition of the eggs, including their fatty acid profile. The results showed that modifying the laying hens’ feed composition resulted in eggs with high nutritional value, with a favorable change in their fatty acid profile. In most cases, the addition of linseed oil or pomegranate seed oil did not affect the physical and chemical properties of the eggs. However, the diet of laying hens had a positive effect on the fatty acid profile of the egg yolk. The presence of conjugated linolenic acid trienes in eggs produced by laying hens fed a modified diet makes them a potential source of these compounds for human consumption.