Cardiovascular Diseases and Marine Oils: A Focus on Omega-3 Polyunsaturated Fatty Acids and Polar Lipids.

Cardiovascular diseases (CVD) remain the leading cause of death across the globe, hence, establishing strategies to counteract CVD are imperative to reduce mortality and the burden on health systems. Dietary modification is an effective primary prevention strategy against CVD. Research regarding dietary supplementation has become increasingly popular. This review focuses on the current in vivo, in vitro, and epidemiological studies associated with that of omega-3 polyunsaturated fatty acids (n-3 PUFAs) and polar lipids (PLs) and how they play a role against CVD. Furthermore, this review focuses on the results of several major clinical trials examining n-3 PUFAs regarding both primary and secondary prevention of CVD. Notably, we place a lens on the REDUCE-IT and STRENGTH trials. Finally, supplementation of PLs has recently been suggested as a potential alternative avenue for the reduction of CVD incidence versus neutral forms of n-3 PUFAs. However, the clinical evidence for this argument is currently rather limited. Therefore, we draw on the current literature to suggest future clinical trials for PL supplementation. We conclude that despite conflicting evidence, future human trials must be completed to confirm whether PL supplementation may be more effective than n-3 PUFA supplementation to reduce cardiovascular risk.

Kombucha Fermentation of Olympus Mountain Tea (Sideritis scardica) Sweetened with Thyme Honey: Physicochemical Analysis and Evaluation of Functional Properties.

This study implemented kombucha fermentation of Olympus Mountain tea (Sideritis scardica) sweetened with honey (OMTWH) in order to investigate the potential for producing a novel beverage with functional properties. The increase in the total count of bacteria and yeast suggests that the OMTWH acts as a viable substrate for supporting the proliferation of the microorganisms of the Kombucha symbiotic culture. The fermentation resulted in a reduction in pH and increased total titratable acidity. After fermentation, a statistically significant increase in the vitamins C, B1, B2, B6, B7, and B12 content was observed (p < 0.05). Total phenolics and antioxidant activity of the fermented beverage was significantly enhanced, as assessed by the method of Folin-Ciocalteu and ABTS assay, respectively. Results revealed that OMTWH had a potent inhibitory activity of α-amylase, α-glucosidase, acetylcholinesterase, and butyrylcholinesterase; OMTWH fermented with a kombucha consortium exhibited even higher inhibition. Hence, the process of kombucha fermentation can convert OMTWH into a novel beverage with enhanced functional properties.

Ultrasonic-Assisted Extraction of Astaxanthin from Shrimp By-Products Using Vegetable Oils.

BACKGROUND: The use of conventional astaxanthin extraction methods, typically involving organic solvents, leads to a heightened environmental impact. The aim of this study was to explore the potential use of environmentally friendly extraction solvents, such as vegetable oils, for recovering the shrimp by-product astaxanthin. METHODS: Ultrasound-assisted extraction (UAE) in vegetable oils, including olive oil (OO), sunflower oil (SO), and flaxseed oil (FO), was employed to extract astaxanthin. The astaxanthin antioxidant activity was evaluated using an ABTS assay, and a mixture of gum Arabic and soy lecithin was used to form coacervates to produce astaxanthin encapsulation. RESULTS: A by-product-vegetable oil ratio of 1:60, extraction time of 210 min, 60% amplitude of the extraction process, and the use of OO as the extracting medium resulted in an astaxanthin yield of 235 ± 4.07 µg astaxanthin/g by-products. The astaxanthin encapsulation efficiency on day 0 and astaxanthin recovery on day 1 were recorded at 66.6 ± 2.7% and 94.4 ± 4.6%, respectively. CONCLUSIONS: The utilization of OO as an extraction solvent for astaxanthin from shrimp by-products in UAE represents a novel and promising approach to reducing the environmental impact of shrimp by-products. The effective astaxanthin encapsulation efficiency highlights its potential application in food industries.

Εvaluation of the microbial stability and shelf life of 50% NaCl-reduced traditional Greek pork meat product "Syglino of Monemvasia" stored under vacuum at different temperatures.

Nowadays, consumers are increasingly concerned about nutrition and health issues. "Syglino of Monemvasia" is a traditional Greek cooked and smoked, sliced pork meat product. Although this is a nutritious food, its consumption should be done in moderation due to the pickling process of its preparation. This product was thus here optimized to contain half salt (NaCl) amount and its microbial stability and shelf life was then assessed in comparison to the already available commercial product. For this, the total viable counts (TVCs) and some critical specific spoilage associations were enumerated at each product type during vacuum incubation at four different temperatures (0, 5, 10, and 15 °C). The alterations in pH, aw, color, and some other crucial sensory attributes of each product were also periodically monitored. The new low-salt product was found to remain microbiologically stable under refrigerated vacuum storage for approximately two weeks, being finally spoiled by Brochothrix thermosphacta grown above 107 CFU/g, ultimately resulting in the deterioration of taste, odor, and overall appearance of the product, and thus leading to its subsequent organoleptic rejection. Despite its limited shelf life, the 50% NaCl-reduced "Syglino" could be released in the local market provided that the cooling chain is maintained throughout its distribution (≤5 °C) and at the same time consumers are willing to accept its milder taste. Although salt replacers could have been used to improve its flavor and at the same time increase its shelf life, the product here developed without the use of such alternatives will hopefully contribute to the taste training of its consumers for less salt in their diet, keeping in parallel its clean label with no added preservatives and other food additives.

Bioactive Lipids of Marine Microalga Chlorococcum sp. SABC 012504 with Anti-Inflammatory and Anti-Thrombotic Activities.

Microalgae are at the start of the food chain, and many are known producers of a significant amount of lipids with essential fatty acids. However, the bioactivity of microalgal lipids for anti-inflammatory and antithrombotic activities have rarely been investigated. Therefore, for a sustainable source of the above bioactive lipids, the present study was undertaken. The total lipids of microalga Chlorococcum sp., isolated from the Irish coast, were fractionated into neutral-, glyco-, and phospho-lipids, and were tested in vitro for their anti-inflammatory and antithrombotic activities. All tested lipid fractions showed strong anti-platelet-activating factor (PAF) and antithrombin activities in human platelets (half maximal inhibitory concentration (IC50) values ranging ~25-200 µg of lipid) with the highest activities in glyco- and phospho-lipid fractions. The structural analysis of the bioactive lipid fraction-2 revealed the presence of specific sulfoquinovosyl diacylglycerols (SQDG) bioactive molecules and the HexCer-t36:2 (t18:1/18:1 and 18:2/18:0) cerebrosides with a phytosphingosine (4-hydrosphinganine) base, while fraction-3 contained bioactive phosphatidylcholine (PC) and phosphatidylethanolamine (PE) molecules. These novel bioactive lipids of Chlorococcum sp. with putative health benefits may indicate that marine microalgae can be a sustainable alternative source for bioactive lipids production for food supplements and nutraceutical applications. However, further studies are required towards the commercial technology pathways development and biosafety analysis for the use of the microalga.

In Vitro Antithrombotic Properties of Salmon (Salmo salar) Phospholipids in a Novel Food-Grade Extract.

Marine and salmon polar lipids (PLs) extracted by conventional extractions with non-food-grade solvents (CE-salmon-PLs) possess antithrombotic bioactivities against platelet-activating factor (PAF) and thrombin. Similar effects of food-grade-extracted (FGE) marine PLs have not yet been reported. In this study, food-grade solvents were used to extract PLs from Irish organic farmed salmon (Salmo salar) fillets (FGE-salmon-PLs), while their antithrombotic bioactivities were assessed in human platelets induced by platelet aggregation agonists (PAF/thrombin). FGE-salmon-PLs were further separated by thin layer chromatography (TLC) into lipid subclasses, and the antithrombotic bioactivities of each subclass were also assessed. LC-MS was utilized to elucidate the structure-activity relationships. FGE-salmon-PLs strongly inhibited PAF-induced platelet aggregation, while their relevant anti-thrombin effects were at least three times more potent than the previously reported activities of CE-salmon-PLs. TLC-derived lipid fractions corresponding to phosphatidylcholines (PC) and phosphatidylethanolamines (PE) were the most bioactive lipid subclasses obtained, especially against thrombin. Their LC-MS analysis elucidated that they are diacyl- or alkyl-acyl- PC and PE moieties baring ω3 polyunsaturated fatty acids (PUFA) at their sn-2 position, such as eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA). Our results concerning the potent antithrombotic effects of FGE-salmon-PLs against both PAF and thrombin pathways strongly suggest that such food-grade extracts are putative candidates for the development of novel cardioprotective supplements and nutraceuticals.

Structural Elucidation of Irish Organic Farmed Salmon (Salmo salar) Polar Lipids with Antithrombotic Activities.

While several marine polar lipids (PL) have exhibited cardioprotective properties through their effects on the platelet-activating factor (PAF) pathways, salmon PL have not been tested so far. In this study, the antithrombotic activities of salmon PL were assessed in human platelets and the structural characterisation of bioactive salmon PL was performed by GC-MS and LC-MS analyses. PL from fillets of Irish organic farmed salmon (Salmo salar) were extracted and separated into several lipid subclasses by thin-layer chromatography (TLC), while their fatty acid profile was fully characterised by GC-MS. Salmon total lipids (TL), total neutral lipids (TNL), total polar lipids (TPL), and each PL subclass obtained by TLC were further assessed for their in vitro effects towards PAF-induced and thrombin-induced platelet aggregation in human platelets. Salmon PL exhibited antithrombotic effects on human platelet aggregation, mostly through their strong inhibitory effects against the PAF pathway with IC50 values comparable to other marine PL, but with lower effects towards the thrombin pathway. PL fractions corresponding to phosphatidylcholine and phosphatidylethanolamine derivatives exhibited the most potent anti-PAF effects, while LC-MS analysis putatively elucidated their structure/function relationship. Several diacyl-PC/PE and alkyl-acyl-PC/PE species containing mostly docosahexaenoic acid at their sn-2 glycerol-backbone may be responsible for the bioactivity. The data presented suggests that salmon contains PL with strong antithrombotic bioactivities.

Evaluation of sensory and in vitro anti-thrombotic properties of traditional Greek yogurts derived from different types of milk.

Given that fermented dairy products exhibit high bioactivities against cardiovascular diseases (CVDs), the anti-thrombotic properties, fatty acid profiles and sensory properties of cow, goat and ewe derived Greek yogurts have been assessed and compared. The total lipids (TL), total polar lipids (TPL), total neutral lipids (TNL) were obtained and the polar lipid fractions were further separated by thin layer chromatography (TLC). These lipid samples (TL, TPL and TLC fractions) were subsequently assessed for their biological activity against atherosclerosis based on the in vitro inhibition of Platelet Activating Factor (PAF)-induced platelet aggregation. The fatty acid compositions of all yogurts were analyzed by Gas Chromatography with flame ionization detector (GC-FID). Goat yogurt lipids have been found to exert more potent inhibitory activity (i.e. lower IC50 values in both TL and TPL samples) in contrast to the corresponding fractions of cow and ewe yogurts. The observed sensory data indicates that ewe yogurt was the most palatable of all three Greek yogurts.

Localization of strawberry (Fragaria x ananassa) and Methylobacterium extorquens genes of strawberry flavor biosynthesis in strawberry tissue by in situ hybridization.

Strawberry flavor is one of the most popular fruit flavors worldwide, with numerous applications in the food industry. In addition, the biosynthetic origin of the most important strawberry flavor components, such as 2,5-dimethyl-4-hydroxy-2H-furan-3-one (DMHF), is a challenging research area. DMHF's precursor, 2-hydroxy-propanal (or lactaldehyde), is biosynthesized by the endophytic bacterium Methylobacterium extorquens (M. extorquens). In particular, the alcohol dehydrogenase (ADH) enzymes of M. extorquens are involved in the biogenesis of DMHF precursors since they have the capacity to oxidize the strawberry-derived 1,2-propanediol to lactaldehyde. In this study, the expression of the endophytic ADH and the plant DMHF biosynthesis genes was examined in the tissues of raw and ripe strawberry receptacles by in situ hybridization. The presence of endophytic bacteria was studied in the same tissues by probes targeting bacterial 16S ribosomal ribonucleic acid. Hybridization signals of probes specific for endophytic ADH and plant DMHF biosynthesis genes, as well as bacteria-specific probes, were detected in the same locations. The probes were localized near the plasma membranes or intercellular spaces of cortical and vascular tissues of the receptacle, and intracellularly in the tissues of achenes. By localizing the expression of the endophytic methanol ADH and plant DMHF biosynthesis genes to the same tissues, we have reinforced our original hypothesis that an intimate symbiotic relationship between strawberry and endophytic cells exists and leads to the biosynthesis of DMHF.

The bioaccumulation and physiological effects of heavy metals in carrots, onions, and potatoes and dietary implications for Cr and Ni: a review.

The bioaccumulation of heavy metals in food tubers (carrots, onions, and potatoes) as a result of polluted irrigation water has been studied in this review paper. Given that heavy metals can cause a considerable oxidative stress, the impact of these metals to the physiology of the plants has also been assessed. The consumption of vegetables cross-contaminated with heavy metals carries a considerable risk for humans (especially for children and pregnant women) and these dietary implications are discussed while European Food Safety Authority has been urged to look into this matter of concern.

Structural elucidation of olive pomace fed sea bass (Dicentrarchus labrax) polar lipids with cardioprotective activities.

The purpose of this study was to structurally characterise the polar lipids of sea bass (Dicentrarchus labrax), fed with an experimental diet containing olive pomace (OP), that exhibit cardioprotective activities. OP has been added to conventional fish oil (FO) feed at 4% and this was the OP diet, having been supplemented as finishing diet to fish. Sea bass was aquacultured using either FO or OP diet. At the end of the dietary experiment, lipids in both samples of fish muscle were quantified and HPLC fractionated. The in vitro cardioprotective properties of the polar lipid fractions, using washed rabbit's platelets, have been assessed and the two most biologically active fractions were further analysed by mass spectrometry. The gas-chromatrograpy-mass spectrometric data shows that these two fractions contain low levels of myristic (14:0), oleic (18:1 cis ω-9) and linoleic acids (18:2 ω-6), but high levels of palmitic (16:0) and stearic acids (18:0) as well as eicosadienoic acid (20:2 ω-6). The first fraction (MS1) also contained significant levels of arachidonic acid (20:4 ω-6) and the omega-3 fatty acids: eicosapentaenoic acid (22:5) and docosahexaenoic acid (22:6). Electrospray-mass spectrometry elucidated that the lipid composition of the two fractions contained various diacyl-glycerophospholipids species, where the majority of them have either 18:0 or 18:1 fatty acids in the sn-1 position and either 22:6 or 20:2 fatty acids in the sn-2 position for MS1 and MS2, respectively. Our research focuses on the structure/function relationship of fish muscle polar lipids and cardiovascular diseases and structural data are given for polar lipid HPLC fractions with strong cardioprotective properties.

Evaluation of the in vitro anti-atherogenic properties of lipid fractions of olive pomace, olive pomace enriched fish feed and gilthead sea bream (Sparus aurata) fed with olive pomace enriched fish feed.

Given the pivotal role of Platelet-Activating-Factor (PAF) in atherosclerosis and the cardio-protective role of PAF-inhibitors derived from olive pomace, the inclusion of olive pomace in fish feed has been studied for gilthead sea bream (Sparus aurata). The aim of the current research was to elucidate the anti-atherogenic properties of specific HPLC lipid fractions obtained from olive pomace, olive pomace enriched fish feed and fish fed with the olive pomace enriched fish feed, by evaluating their in vitro biological activity against washed rabbit platelets. This in vitro study underlines that olive pomace inclusion in fish feed improves the nutritional value of both fish feed and fish possibly by enriching the marine lipid profile of gilthead sea bream (Sparus aurata) with specific bioactive lipid compounds of plant origin.

Hen egg yolk lipid fractions with antiatherogenic properties.

Three different types of hen egg yolk, cage-free, organic and daily fresh, were tested for their antiatherogenic properties. Total lipids (TL) of all hen egg yolk samples were extracted by the method of Bligh and Dyer and further separated into total polar lipids (TPL) and total neutral lipids (TNL) by counter current distribution chromatography. TPL and TNL were further separated by preparative thin-layer chromatography (TLC). TL, TPL, TNL and the obtained polar and neutral lipid fractions after TLC separation were tested to determine whether they induce platelet activation or inhibit platelet activating factor (PAF)-induced platelet activation. All three hen egg yolk TL samples possessed strong inhibitory activity against PAF-induced platelet activation that was mainly attributed to TPL, especially to PL fraction 4. Cage-free hen egg yolk exhibited the most potent anti-PAF activity in all lipid classes (TL, TPL and TNL). Thus hen egg yolk contains PAF inhibitors that reinforce their nutritional value in terms of protection against cardiovascular diseases, since PAF is a crucial inflammatory phospholipid mediator that is implicated in the mechanism of atherogenesis.

Fish polar lipids retard atherosclerosis in rabbits by down-regulating PAF biosynthesis and up-regulating PAF catabolism.

BACKGROUND: Platelet activating factor (PAF) has been proposed as a key factor and initial trigger in atherosclerosis. Recently, a modulation of PAF metabolism by bioactive food constituents has been suggested. In this study we investigated the effect of fish polar lipid consumption on PAF metabolism. RESULTS: The specific activities of four PAF metabolic enzymes; in leukocytes, platelets and plasma, and PAF concentration; either in blood cells or plasma were determined. Samples were acquired at the beginning and at the end of a previously conducted study in male New Zealand white rabbits that were fed for 45 days with atherogenic diet supplemented (group-B, n = 6) or not (group-A, n = 6) with gilthead sea bream (Sparus aurata) polar lipids.The specific activity of PAF-Acetylhydrolase (PAF-AH); a catabolic enzyme of PAF, was decreased in rabbits' platelets of both A and B groups and in rabbits' leukocytes of group A (p < 0.05). On the other hand the specific activity of Lipoprotein-associated Phospholipase A2 (Lp-PLA2); the catabolic enzyme of PAF in plasma was increased in both A and B groups in both leukocytes and platelets (p < 0.05). PAF-cholinephosphotransferase (PAF-CPT); a biosynthetic enzyme of PAF showed increased specific activity only in rabbits' leukocytes of group A (p < 0.05). Neither of the two groups showed any change in Lyso-PAF-acetyltransferase (Lyso-PAF-AT) specific activity (p > 0.05). Free and bound PAF levels increased in group A while decreased in group B (p < 0.05). CONCLUSIONS: Gilthead sea bream (Sparus aurata) polar lipids modulate PAF metabolism upon atherosclerotic conditions in rabbits leading to lower PAF levels and activity in blood of rabbits with reduced early atherosclerotic lesions compared to control group.

Effects of olive pomace and olive pomace oil on growth performance, fatty acid composition and cardio protective properties of gilthead sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax).

Gilthead sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax) were fed with two experimental diets: olive pomace diet and olive pomace oil diet, in order to examine whether fish oil substitution in fish feeds by olive oil production wastes, such as olive pomace and olive pomace oil, has an influence on growth performance, fatty acid composition and cardio protective properties of fish. Gilthead sea bream fed with both experimental diets exhibited satisfactory growth performance factors while sea bass did not. The total lipids of gilthead sea bream fed with olive pomace diet contained statistically decreased levels of fatty acids, while exhibited the most potent biological activity against platelet aggregation induced by Platelet Activating Factor. These data indicate that olive pomace can be used as a partial substitute of fish oil in fish feed improving its cardio protective properties.

Antibacterial and anti-PAF activity of lipid extracts from sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata).

The anti-PAF and the antibacterial activities of lipid extracts obtained from cultured sea bass (Dicentrarchus labrax) and cultured gilthead sea bream (Sparus aurata) were evaluated. Total lipids of sea bass and gilthead sea bream exerted PAF-like activity while, in higher amounts they inhibited this PAF activity. Neutral lipids of both sea bass and gilthead sea bream contained only PAF antagonists while the polar lipid fractions contained both PAF antagonists and agonists. Total lipids of sea bass exhibited stronger PAF-like activity than did those of gilthead sea bream; however, neutral lipids of sea bass contained stronger PAF antagonists than did gilthead sea bream. Total lipids of both sea bass and gilthead sea bream exhibited antibacterial activity only towards Staphylococcus aureus (S. aureus) with those of sea bass being more potent. Subsequently, neutral lipids of both sea bass and gilthead sea bream also showed antibacterial activity against S. aureus and less so towards Escherichia coli (E. coli), while only neutral lipids of sea bass showed antibacterial activity against Enterococcusfaecalis (E. faecalis). Sea bass neutral lipids were more active against S. aureus than were those of gilthead sea bream, while their activity towards E. coli was similar. Polar lipids of both sea bass and gilthead sea bream showed antibacterial activity against all bacteria strains. Sea bass polar lipids were more active towards S. aureus than were those of gilthead sea bream, while their activities against E. faecalis and E. coli were the same. The detected antibacterial activities of the lipid extracts isolated from sea bass and gilthead sea bream were observed in amounts equal to those that exerted either PAF inhibition or PAF-like activity, suggesting that PAF antagonists and agonists of fish lipids may be responsible for the antibacterial activity.