Effect of phosphatidylcholine regioisomerism on lateral segregation of milk sphingomyelin in bilayer membranes.

Milk fat globule membrane (MFGM) promotes the lateral phase separation of milk lipids and stabilizes the fat globules in milk. The composition and structures of lipids have a significant impact on physicochemical properties of MFGM, which in turn influences the digestion and absorption of milk lipids. Phospholipids (PL), sphingolipids, and cholesterol are the major lipid constituents of MFGM. While the effects of the head-group and structure of the fatty acids (FAs) on membrane properties are commonly studied, little is known on the impact of PL regioisomerism. The present study investigated the impact of phosphatidylcholine (PC) regioisomerism on lateral segregation of milk-sphingomyelin (milk-SM) as well as the influence on the interaction of milk-SM with ceramide and cholesterol in simulated membrane systems. The regioisomer pairs of four molecular species PC 16:0/18:1n-9, PC 16:0/18:2n-6, PC 16:0/18:3n-3, and PC 16:0/20:4n-6 were included in this study. The lateral segregation was determined using lifetime analysis of trans-parinaric acid (tPA) fluorescence. Thermostability of the domains was detected using steady-state anisotropy of tPA. Our results demonstrated a clear impact of PC regioisomerism on membrane properties. PC regioisomers having the unsaturated FAs at the sn-2 position enhanced the lateral segregation of milk-SM with and without the presence of ceramide and cholesterol compared to the regioiosmers having 16:0 at the sn-2 position. Furthermore, the characteristics i. e. the acyl chain length and degree of unsaturation of sn-2 FA of the PCs had a major impact on the milk-SM gel phase and the intermolecular forces between milk-SM and ceramide/cholesterol. This work is the first investigation showing the effect of PL regioisomerism on milk-SM domains, which might have significant influence on functional properties of MFGM.

Enantiomeric Separation of Triacylglycerols Consisting of Three Different Fatty Acyls and Their Chiral Chromatographic Elution Behavior.

Chromatographic separation of triacylglycerol (TG) enantiomers is a highly challenging task of analytical chemistry because of the similar physicochemical properties. The analysis of chiral TGs is crucial for improving the knowledge of lipid biochemistry and for understanding the nutritional properties of fats and oils. Thus, this study aimed to systematically investigate the chiral resolution of TGs consisting of three different fatty acyls (FAs). Thirty-three asymmetric TG enantiopairs, including 49 synthesized enantiopure TGs and racemic TGs, were analyzed with a recycling chiral HPLC system. Twenty-six enantiopairs were successfully separated. Overall, having both unsaturated and saturated FAs in the outer positions or a difference in carbon chain length between two saturated FAs at the outer positions favored the separation of enantiomers. The retention time at separation correlated negatively with the sn-3 carbon number of the early eluting enantiomer and positively with the carbon number difference between sn-1 and sn-3. When the samples were studied in separate groups based on unsaturation and regioisomers, both the acyl carbon number and the degree of unsaturation of FAs in all three positions influenced the separation and elution behavior of chiral TGs, indicating an active role of both intermolecular interactions and steric hindrances. This is the first systematic study of the chiral separation of TGs consisting of three different FAs using a large number of enantiopairs. The novel findings on the behavior of TG enantiomers in a chiral environment provide important guidance and reference for a stereospecific study of the chemistry and biochemistry of natural lipids.

Enzymatic acylation of cyanidin-3-O-glucoside with aromatic and aliphatic acid methyl ester: Structure-stability relationships of acylated derivatives.

Anthocyanins are water-soluble pigments, but they tend to be unstable in aqueous solutions. Modification of their molecular structure offers a viable approach to alter their intrinsic properties and enhance stability. Aromatic and aliphatic acid methyl esters were used as acyl donors in the enzymatic acylation of cyanidin-3-O-glucoside (C3G), and their analysis was conducted using ultraperformance liquid chromatography-mass spectrometry (UPLC-MS). The highest conversion rate achieved was 96.41 % for cyanidin-3-O-(6″-feruloyl) glucoside. Comparative evaluations of stability revealed that aromatic acyl group-conjugated C3G exhibited superior stability enhancement compared with aliphatic acyl group derivatives. The stability of aliphatic C3G decreased with increasing carbon chain length. The molecular geometries of different anthocyanins were optimized, and energy level calculations using density functional theory (DFT) identified their sites with antioxidant activities. Computational calculations aligned with the in vitro antioxidant assay results. This study provided theoretical support for stabilizing anthocyanins and broadened the application of acylated anthocyanins as food colorants and nutrient supplements.

Reduced seed viability in exchange for transgenerational plant protection: Does the defensive mutualism concept pass the fitness test?

BACKGROUND AND AIMS: Epichloë endophytes are vertically transmitted via grass seeds and chemically defend their hosts against herbivory. Endophyte-conferred plant defence via alkaloid biosynthesis may occur independently of costs for host plant growth. However, fitness consequences of endophyte-conferred defence and transgenerational effects on herbivore resistance of progeny plants, are rarely studied. The aim of this study was to test whether severe defoliation in mother plants affects their seed production, seed germination rate, and the endophyte-conferred resistance of progeny plants. METHODS: In a field study, we tested the effects of defoliation and endophyte symbiosis (Epichloë uncinata) on host plant (Festuca pratensis) performance, loline alkaloid concentrations in leaves and seeds, seed biomass and seed germination rates. In a subsequent greenhouse study, we challenged the progeny of the plants from the field study to aphid herbivory and tested whether defoliation of mother plants affects endophyte-conferred resistance against aphids in progeny plants. KEY RESULTS: Defoliation of the mother plants resulted in a reduction of alkaloid concentrations in leaves and elevated the alkaloid concentrations in seeds when compared with non-defoliated endophyte-symbiotic plants. Viability and germination rate of seeds of defoliated endophyte-symbiotic plants were significantly lower compared to those of non-defoliated endophyte-symbiotic plants and endophyte-free (defoliated and non-defoliated) plants. During six weeks growth, seedlings of defoliated endophyte-symbiotic mother plants had elevated alkaloid concentrations, which negatively correlated with aphid performance. CONCLUSIONS: Endophyte-conferred investment in higher alkaloid levels in seeds -elicited by defoliation- provided herbivore protection in progenies during the first weeks of plant establishment. Better protection of seeds via high alkaloid concentrations negatively correlated with seed germination indicating trade-off between protection and viability.

Synergistic effect of bath-ultrasonication and heating treatments on two-steps treatment of brewers' spent grain.

The study aimed to evaluate the chemical composition, antioxidant activity and techno-functionality of brewers' spent grain (BSG) treated with two-steps treatment involving 5, 15, and 25 min bath-ultrasonication (USB) continued with autoclave (AH) at 90, 110, and 130 °C and/or water-bath (CWH) at 80, 90, and 100 °C. The two-steps treatments slightly affected the water- and oil-holding capacity and extractable fat content. Most of the two-steps treatments increased the amount of flavan-3-ols and phenolic acids, up to 4 times higher compared to its control. The two-steps treatment involving CWH had no significant (p > 0.05) impact on fat content, antioxidants and techo-functionality of BSG. Up to 15 min USB increased the poly-unsaturated fatty acids and lowered the amount of saturated fatty acids. In conclusion, the two-steps treatment consists of USB (up to 15 min) continued with AH and CWH increased the amount of nutritional-related chemical composition such as UFA and phenolic acids as well as antioxidant activity of BSG.

Anthocyanins as natural bioactives with anti-hypertensive and atherosclerotic potential: Health benefits and recent advances.

BACKGROUND: Hypertension is a highly prevalent chronic metabolic illness affecting individuals of all age groups. Furthermore, it is a significant risk factor for the development of atherosclerosis (AS), as a correlation between hypertension and AS has been observed. However, the effective treatments for either of these disorders appear to be uncommon. METHODS: A systematic search of articles published in PubMed, Web of Science, ScienceDirect, Scopus, and Google Scholar databases over the last decade was performed using the following keywords: hypertension, AS, anthocyanins, antioxidants, gut microbes, health benefits, and bioactivity. RESULTS: The available research indicates that anthocyanin consumption can achieve antioxidant effects by inducing the activation of intracellular nuclear factor erythroid 2-related factor (Nrf2) and the expression of antioxidant genes. Moreover, previous reports showed that anthocyanins can enhance the human body's ability to fight against inflammation and cancer through the inhibition of inflammatory factors and the regulation of related signaling pathways. They can also protect the blood vessels and nervous system by regulating the production and function of endothelial nitric oxide synthase (eNOS). Gut microorganisms play an important role in various chronic diseases. Our research has also investigated the role of anthocyanins in the metabolism of the gut microbiota, leading to significant breakthroughs. This study not only presents a unique strategy for reducing the risk of cardiovascular diseases (CVDs) without the need for medicine but also provides insights into the development and utilization of intestinal probiotic dietary supplements. CONCLUSION: In this review, different in vitro and in vivo studies have shown that anthocyanins slow down the onset and progression of hypertension and AS through different mechanisms. In addition, gut microbial metabolites also play a crucial role in diseases through the gut-liver axis.

Chemoenzymatic Synthesis of ABC-Type Enantiostructured Triacylglycerols by the Use of the p-Methoxybenzyl Protective Group.

This report demonstrates the first asymmetric synthesis of enantiopure structured triacylglycerols (TAGs) of the ABC type presenting three non-identical fatty acids, two of which are unsaturated. The unsaturated fatty acids included monounsaturated oleic acid (C18:1 n-9) and polyunsaturated linoleic acid (C18:2 n-6). This was accomplished by a six-step chemoenzymatic approach starting from (R)- and (S)-solketals. The highly regioselective immobilized Candida antarctica lipase (CAL-B) played a crucial role in the regiocontrol of the synthesis. The synthesis also benefited from the use of the p-methoxybenzyl (PMB) ether protective group, which enabled the incorporation of two different unsaturated fatty acids into the glycerol skeleton. The total of six such TAGs were prepared, four constituting the unsaturated fatty acids in the sn-1 and sn-2 positions, with a saturated fatty acid in the remaining sn-3 position of the glycerol backbone. In the two remaining TAGs, the different unsaturated fatty acids accommodated the sn-1 and sn-3 end positions, with the saturated fatty acid present in the sn-2 position. Enantiopure TAGs are urgently demanded as standards for the enantiospecific analysis of intact TAGs in fats and oils.

Advanced Tandem Mass Spectrometric Analysis of Complex Mixtures of Triacylglycerol Regioisomers: A Case Study of Bovine Milk Fat.

Comprehensive analysis of triacylglycerol (TAG) regioisomers is extremely challenging, with many variables that can influence the results. Previously, we reported a novel algorithmic method for resolving regioisomers of complex mixtures of TAGs. In the current study, the TAG Analyzer software and its mass spectrometric fragmentation model were further developed and validated for a much wider range of TAGs. To demonstrate the method, we performed for the first time a comprehensive analysis of TAG regioisomers of bovine milk fat, a very important and one of the most complex TAG mixtures in nature containing FAs ranging from short to long carbon chains. This analysis method forms a solid basis for further investigation of TAG regioisomer profiles in various natural fats and oils, potentially aiding in the development of new and healthier foods and nutraceuticals with targeted lipid structures.

Growth conditions trigger genotype-specific metabolic responses that affect the nutritional quality of kale cultivars.

Kales (Brassica oleracea convar acephala) are fast-growing, nutritious leafy vegetables ideal for year-round indoor farming. However, selection of best cultivars for growth under artificial lighting necessitates a deeper understanding of leaf metabolism in different kale types. Here we examined a curly leaved cultivar Half Tall and a lacinato type cultivar Black Magic under moderate growth light (130 µmol photons m-1s-1/22°C) and high light (800 µmol photons m-1s-1/26°C) conditions. These conditions induced genotype-dependent differences in nutritionally important metabolites, especially anthocyanins and glucosinolates (GSLs), in the kale cultivars. In the pale green Half Tall, growth under high light conditions did not induce changes in either pigmentation or total GSL content. In contrast, the purple pigmentation of Black Magic intensified due to increased anthocyanin accumulation. Black Magic showed reduced amounts of indole GSLs and increased amounts of aliphatic GSLs under high light conditions, with notable cultivar-specific adjustments in individual GSL species. Correlation analysis of metabolite profiles suggested cultivar-specific metabolic interplay between serine biosynthesis and the production of indole GSLs. RNA sequencing identified candidate genes encoding metabolic enzymes and regulatory components behind anthocyanin and GSL biosynthesis. These findings improve the understanding of leaf metabolism and its effects on the nutritional quality of kale cultivars.

Lipid and volatile profiles of Finnish oat batches of pure cultivars: Effect of storage on the volatile formation.

Recent data showing the compositional variation and storage behavior among different oat batches for the purpose of food remains limited. Lipids of twenty oat flour samples of pure cultivars grown in Finland during 2019 were extracted and fractionated into neutral and polar-rich lipids. Flour was stored for nine months, and profiles of volatiles and tocols were analyzed to reveal oxidative stability. The lipid content was 5.9-8.9 g per 100 g of flour [DW] and consisted of 78.7 ± 2.5 % neutral and 21.3 ± 2.5 % polar lipids. Palmitic (16 %), oleic (36 %), and linoleic (39 %) acids were the most abundant fatty acids. Neutral lipids had more oleic and less linoleic and palmitic acids than polar lipids. The fresh samples correlated with tocols, pentanal, 2-pentylfuran, 2-heptanone, nonanal, 2-butanone, and heptanal, while stored samples were associated with 3-octen-2-one, 2-octenal, hexanal, and octanal. Lipid composition and oxidative stability are essential factors for selecting oat batches for food applications.

Influence of Dietary Triacylglycerol Structure on the Accumulation of Docosahexaenoic Acid [22:6(n-3)] in Organs in a Short-Term Feeding Trial with Mildly Omega-3 Deficient Rats.

SCOPE: To study the effect of positional distribution of docosahexaenoic acid (DHA) in dietary triacylglycerols (TAG) on the tissue fatty acid content and composition of mildly (n-3) deficient rats. METHODS AND RESULTS: In a 5-day feeding trial, mildly (n-3) deficient rats received 360 mg daily structured TAGs: sn-22:6(n-3)-18:0-18:0, sn-18:0-18:0-22:6(n-3), sn-18:0-22:6(n-3)-18:0, or tristearin. A fifth group receives standard (n-3) adequate feed AIN-93G from birth till the end of the trial. The DHA-fed groups show significantly higher DHA levels in the liver and visceral fat compared to the tristearin or normal feed groups showing that the dose and the short feeding period of DHA were sufficient to restore the DHA content in the organs of (n-3) deficient rats. Feeding sn-1 DHA resulted in higher levels of DHA in the liver TAG compared to sn-3 DHA feeding, although the difference did not reach statistical significance. CONCLUSION: These findings indicated a possible difference in the tissue accumulation and/or metabolic fate of DHA from the sn-1 and sn-3 positions of TAG.

Selecting autochthonous lactic acid bacteria for co-inoculation in Chinese bayberry wine production: Stress response, starter cultures application and volatilomic study.

This study focused on isolating and characterising autochthonous lactic acid bacteria (LAB) from spontaneously fermented Chinese bayberry (CB) and their potential application in CB wine fermentation in co-inoculation with yeast starter cultures. Numerous LAB, including Lactiplantibacillus (Lp.) plantarum (9), Limosilactobacillus (Lb.) fermentum (6), Lactococcus (Lc.) lactis (3), Enterococcus (Ec.) hirae (1), Leuconostoc (Le.) mesenteroides (1), and Weissella (Ws.) cibaria (1), were isolated and identified. The isolated strains Lp. plantarum ZFM710 and ZFM715, together with Lb. fermentum ZFM720 and ZFM722, adapted well to unfavourable fermentation environment, including ethanol, osmolality, and acidity stresses, were selected for producing CB wine by co-inoculation with Saccharomyces cerevisiae. During fermentation, the presence of LAB promoted the development of S. cerevisiae, while the population dynamics of LAB in different groups at different stages showed strain-specific differences. Fermentation trials involving LAB yielded a lower ethanol concentration except for Lp. plantarum ZFM715. Compared to the pure S. cerevisiae fermented sample, the addition of LAB led to a clear modulation in organic acid composition. Lb. fermentum strains in co-fermentation led to significant decreases in each classified group of aroma compounds, while Lp. plantarum ZFM715 significantly increased the complexity and intensity of aroma compounds, as well as the intensities of fruity and floral notes. The study selects interesting strains for the design of starter cultures for use in CB wine production, underlining the interest in the selection of autochthonous LAB in fruit wines, with the aim of improving the adaptation of bacteria to specific environmental conditions and shaping the unique traits of the finished products.

Metabolomic Investigation of Brain and Liver in Rats Fed Docosahexaenoic Acid in Regio- and Enantiopure Triacylglycerols.

SCOPE: N-3 polyunsaturated fatty acids (n-3 PUFAs) play important roles in cognitive functions. However, there is a lack of knowledge on the metabolic impact of regio- and stereo-specific positioning of n-3 PUFAs in dietary triacylglycerols. METHODS AND RESULTS: Rats in a state of mild n-3 PUFA deficiency are fed daily with 360 mg triacylglycerols containing DHA (docosahexaenoic acid) at sn (stereospecific numbering)-1, 2, or 3 positions and 18:0 at remaining positions, or an equal amount of tristearin for 5 days. Groups fed with n-3 deficient diet and normal n-3 adequate diet are included as controls. The metabolic profiles of the brain and liver are studied using NMR (nuclear magnetic resonance)-based metabolomics. Several metabolites of significance in membrane integrity and neurotransmission, and glutamate, in particular, are significantly lower in the brain of the groups fed with sn-1 and sn-3 DHA compared to the sn-2 DHA group. Further, the tristearin and DHA groups show a lower lactate level compared to the groups fed on normal or n-3 deficient diet, suggesting a prominent role of C18:0 in regulating energy metabolism. CONCLUSION: This study sheds light on the impact of stereospecific positioning of DHA in triacylglycerols and the role of dietary stearic acid on metabolism in the brain and liver.

Flavonoid diversity in bitter and debittered seeds of Andean lupin (Lupinus mutabilis Sweet).

Seeds of ten Andean lupin (Lupinus mutabilis Sweet) ecotypes were collected from different regions of Peru and treated with an aqueous debittering method. Both untreated and treated seeds were analyzed by using LC-MS to investigate flavonoid profiles of different ecotypes and impact of debittering process on these compounds. Thirteen isoflavones (mainly as glycosides of genistein and methoxy-genistein) and eight flavones (glycosylated apigenins and methyl-luteolins) were characterized as the main flavonoids in the seed samples. The untreated lupin seeds contained 187-252 mg/100 g (dry weight) of flavonoids. The main difference among lupin ecotypes was observed in the levels of genistein-malonylhexoside, methoxy-genistein-malonylhexoside, and methyl-luteolin-malonylhexoside. After the debittering treatment, the total flavonoid content in the seeds was decreased to 125-203 mg/100 g dry weight, the aglycones of genistein, methoxy-genistein, and methyl-luteolin being the key distinguishing compounds of ecotypes. The aqueous treatment was effective in degrading flavonoid glycosides and releasing the corresponding aglycones.

Metabolic fate of DHA from regio- and stereospecific positions of triacylglycerols in a long-term feeding trial in rats.

This study investigated the impact of regio- and stereospecific position of docosahexaenoic acid (DHA) in dietary triacylglycerols (TAGs) on the fatty acid composition of tissues and organs in rats. Four-week feeding with TAGs containing DHA in sn-1, 2, or 3 position and palmitic acid in the remaining positions at a daily dosage of 500 mg TAG/kg body weight significantly increased the DHA content in all organs and tissues in rats, except in the brain, where the change in DHA level was not statistically significant. The group fed sn-1 DHA showed a significantly higher content of DHA in the plasma TAG than the group fed sn-3 DHA. The sn-3 DHA group had higher levels of DHA in the visceral fat compared to the sn-1, sn-2, as well as all other groups. This is the first study showing that DHA from sn-1 and sn-3 positions of dietary TAGs have differential accumulation in tissues. The new findings improved the current knowledge on the significance of TAG isomeric structure for the bioavailability and metabolic fate of DHA.

Correction: Kulkarni et al. Tissue-Specific Content of Polyunsaturated Fatty Acids in (n-3) Deficiency State of Rats. Foods 2021, 11, 208.

The authors would like to make the following corrections to a published paper [...].

Campaign education and communication to the potential consumers of brewers' spent grain (BSG)-added food products as sustainable foods.

Byproduct of the brewery industry, brewers' spent grain (BSG), has been reported for enhancing the nutritional value of food products thus potentially as a future sustainable food ingredient. As it is a byproduct, society as potential consumers tend to consider BSG as waste. Consequently, commercialization of BSG-added food products would be responded negatively by potential consumers. Campaign education seems to be important in order to familiarize such products to the society thus change the consumers perspective and finally increase the consumers acceptability. This study, which was conducted by an online survey platform (surveymonkey.com), aims to educate the society about the positive value of BSG for human health and its valorisation to the environment which generates sustainable food and to observe their opinion and expectation regarding such products. Majority of participants (57.38%) never knew BSG before. After BSG was introduced, most of the provided information about BSG is new for the majority of participants and they are convinced that consuming BSG-added food products would be beneficial for their health and help a sustainable environment. Furthermore, if such products are claimed as healthy foods, 81.97% of the participants are willing to buy. Besides the mindset of consumers, several issues might need to be considered in commercializing BSG-added food products including food regulation and safety as well as specific health conditions. Further study is still needed in order to educate society in a wider and to be specific in certain groups of community.

Conventional water bath heating on undried brewer's spent grain: Functionality, fatty acids, volatiles, polyphenolic and antioxidant properties.

Brewers' spent grain (BSG) contains bioactive compounds. It was hypothesized that heating treatments using conventional water bath heating (CWH) on brewers' spent grain (BSG) would modify the functionality, chemical constituents and antioxidant activities of BSG. Different temperatures and time exposures (80, 90 and 100 °C at 15, 30 and 60 min) were applied on fresh undried BSG. CWH at 80 °C increased the amount of flavan-3-ols, while 100 °C at 30 and 60 min improved the ABTS (2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) value. CWH significantly declined saturated fatty acid and enhanced the poly-unsaturated fatty acid. Moreover, CWH discharged pungent, floral, spice and mushroom odor perceptions and formed fruity, sweet and pleasant odor perceptions, as well as essential-oil-related compounds. Additionally, CWH improved water-holding and oil-holding capacities. In conclusion, CWH as a low-cost treatment improved the functionality, fatty acid composition and aromatic profile of BSG.

Composition of Sugars, Organic Acids, Phenolic Compounds, and Volatile Organic Compounds in Lingonberries (Vaccinium vitis-idaea L.) at Five Ripening Stages.

Wild lingonberries are a traditional source of food in the Nordic countries and an important contributor to economic activity of non-wood forest products in the region. Lingonberries are a rich source of bioactive compounds and can be a valuable contributor to a healthy diet. However, there are few studies available on how the bioactive compounds in lingonberries develop as they ripen. In this investigation, we examined the content of 27 phenolic compounds, three sugars, four organic acids, and 71 volatile organic compounds at five ripening stages. The study showed that, while the highest content of phenolic compounds was found early in the development, the organoleptic quality of the fruits improved as they ripened. From the first to the last stage of development, anthocyanins went from being nearly absent to 100 mg/100 g fw, and there was an increased content of sugars from 2.7 to 7.2 g/100 g fw, whereas the content of organic acids decreased from 4.9 to 2.7 g/100 g fw, and there were several changes in the profile of volatiles. The contents of flavonols, cinnamic acid derivatives, flavan-3-ols, and the total concentration of phenolic compounds were significantly lower in the fully ripe berries compared to berries in the early green stage. In addition to the changes occurring due to ripening, there was observed variation in the profile of both phenolic compounds and volatiles, depending on the growth location of the berries. The present data are useful for the assessment of harvest time to obtain the desired quality of lingonberries.