Comparison of the effect of various sources of saturated fatty acids on infant follow-on formulas oxidative stability and nutritional profile.

Fortification of infant follow-on formulas (IFF) with docosahexaenoic acid (DHA), which is prone to lipid oxidation, is required by European regulation. This study aimed to identify lipid formulation parameters that improve the nutritional profile and oxidative stability of IFF. Model IFF were formulated using different lipid and emulsifier sources, including refined (POM) or unrefined red palm oil (RPOM), coconut oil (COM), dairy fat (DFOM), soy lecithin, and dairy phospholipids (DPL). After an accelerated storage, RPOM and DFOM with DPL had improved oxidative stability compared to other IFF. Specifically, they had a peroxide value twice lower than POM and 20% less loss of tocopherols for DFOM-DPL. This higher stability was mainly explained by the presence of compounds such as carotenoids in RPOM and sphingomyelin in DFOM-DPL very likely acting synergistically with tocopherols. Incorporation of dairy lipids and carotenoids into DHA-enriched IFF compositions seems promising to enhance their stability and nutritional quality.

Tocopherols as antioxidants in lipid-based systems: The combination of chemical and physicochemical interactions determines their efficiency.

Lipid oxidation is a major concern in the food, cosmetic, and pharmaceutical sectors. The degradation of unsaturated lipids affects the nutritional, physicochemical, and organoleptic properties of products and can lead to off-flavors and to the formation of potentially harmful oxidation compounds. To prevent or slow down lipid oxidation, different antioxidant additives are used alone or in combination to achieve the best possible efficiency with the minimum possible quantities. In manufactured products, that is, heterogeneous systems containing lipids as emulsions or bulk phase, the efficiency of an antioxidant is determined not only by its chemical reactivity, but also by its physical properties and its interaction with other compounds present in the products. The antioxidants most widely used on the industrial scale are probably tocopherols, either as natural extracts or pure synthetic molecules. Considerable research has been conducted on their antioxidant activity, but results regarding their efficiency are contradictory. Here, we review the known mechanisms behind the antioxidant activity of tocopherols and discuss the chemical and physical features that determine their efficacy. We first describe their chemical reactivity linked with the main factors that modulate it between efficient antioxidant capacity and potential prooxidant effects. We then describe their chemical interactions with other molecules (phenolic compounds, metals, vitamin C, carotenes, proteins, and phospholipids) that have potential additive, synergistic, or antagonist effects. Finally, we discuss other physical parameters that influence their activity in complex systems including their specific interactions with surfactants in emulsions and their behavior in the presence of association colloids in bulk oils.

Stability to oxidation and interfacial behavior at the air/water interface of minimally-processed versus processed walnut oil-bodies.

Oil bodies (OB), the form of triacylglycerol storage in seeds, are interesting natural assemblies for nutritional applications. In walnuts, OB contain an important amount of polyunsaturated fatty acids that could be interesting food ingredients but may be prone to oxidation. The oxidative and interfacial behavior of walnut OB, either minimally-processed or after processing, were compared with processed complex walnut juice. The good oxidative stability of minimally-processed OB over 10 days (PV ≤ 8.4 meq O2/kg, TBARS = 1.4 mmol eq MDA/kg) and of processed walnut complex matrixes over 20 days (PV ≤ 4.8 meq O2/kg, TBARS = 1.4 mmol eq MDA/kg) was evidenced. In comparison, processing of OB promoted their oxidation. The interfacial studies led to the proposition of a new model of adsorption for minimally-processed OB that will be useful to design functional emulsion or foam in which OB act as emulsifiers.

Investigation on the Double CutOff Phenomenon Observed in Protocatechuic Acid and Its Alkyl Esters under Various CAT-Based Assays.

A strange cutoff phenomenon of a series of protocatechuic acid alkyl esters had been noticed using the conjugated autoxidizable triene (CAT) assay. Two parabolic shapes of antioxidant activities of protocatechuic acid alkyl esters described as ″the double cutoff effect″ have been speculated as a result of an oxidative driving force generated in the aqueous phase. The aim of this research was to investigate the double cutoff effect using various types of oxidation driving forces in different CAT-based assays. To further explain the phenomenon, the natural oxidation of conjugated autoxidizable triene (NatCAT) assay has been developed for the first time by relying solely on only the lipid autoxidation of tung oil-in-water (O/W) emulsions. In conclusion, NatCAT exhibited different antioxidant and oxidation patterns from both CAT and apolar radical-initiated CAT assays, and only one cutoff point was obtained. This discovery would lead to a greater understanding of the complexity of antioxidant/lipid oxidation dynamics in O/W emulsion systems.

Effect of sinapic acid ester derivatives on the oxidative stability of omega-3 fatty acids rich oil-in-water emulsions.

Oil-in-water (O/W) emulsions are important delivery systems of omega-3 fatty acids (n-3 FA). We investigated the effect of sinapic acid esters concentration and chain length, the electrical charge of the emulsifier and emulsion pH on the oxidative stability of n-3 FA rich O/W emulsions. Echium oil was applied as n-3 FA source. A 24 factorial design was used to simultaneously evaluate these factors. Peroxide value, malondialdehyde, 2,4-heptadienal and 2,4-decadienal were measured in the emulsions. pH and the electrical charge of the emulsifier modulated the antioxidant effectiveness of sinapic acid esters, while concentration was not relevant. The combination of positively charged emulsifier with neutral pH provided the best oxidative stability for echium oil emulsions. Our results also suggested that the increase of length chain of sinapic acid, from C4 to C12, reduced the secondary products of oxidation, when echium oil emulsions were prepared using negatively charged emulsifier under acidic conditions.

Resveratrol-Linoleate protects from exacerbated endothelial permeability via a drastic inhibition of the MMP-9 activity.

Gelatinolytic matrix metalloproteinases (MMP-2, -9) play a critical role not only in mammals physiology but also during inflammation and healing processes. The natural stilbenoid, resveratrol (RES), exhibits potent antioxidant effects, in a hormetic mode of action, and is known to inhibit MMP-9. However, RES administration exhibits major issues, including poor bioavailability and water solubility, hampering its potential therapeutic effect in vivo In the present study, we synthesized and evaluated five novel RES-lipid conjugates to increase their cell membrane penetration and improve their bioavailability. The best in vitro MMP-9 inhibitory activity of RES-lipids conjugates was observed with RES-linoleic acid (LA) (5 µM), when dissolved in a natural deep eutectic solvent (NADES), composed of an equimolar content of 1,2-propanediol:choline chloride (ChCl):water. The inhibition of MMP-9 expression by RES-LA in activated THP-1 monocytes, was, at least due to the deactivation of ERK1/2 and JNK1/2 MAP kinase signaling pathways. Moreover, RES-LA exhibited a strong effect protecting the TNF-α-induced exacerbated permeability in an HUVEC in vitro monolayer (by 81%) via the integrity protection of intercellular junction proteins from the MMP-9 activity. This effect was confirmed by using several complementary approaches including, the real-time monitoring of trans-endothelial electric resistance (TEER), the Transwell HUVEC permeability level, the microscopic examination of the platelet endothelial cell adhesion molecule-1 (CD31/PECAM-1) integrity as well as the fluorescence in intercellular spaces. Consequently, following this strong in vitro proof-of-concept, there is a need to test this promising RES-lipid derivative compound to control the pathological endothelial permeability in vivo.

Eleostearic phospholipids as probes to evaluate antioxidants efficiency against liposomes oxidation.

Regardless of the applications: therapeutic vehicle or membrane model to mimic complex biological systems; it is of a great importance to develop simplified, reproducible and rapid model assays allowing for a relevant assessment of the liposomal membrane oxidation and therefore antioxidant activity of selected molecules. Here, we describe a new and high-throughput assay that we called "Vesicle Conjugated Autoxidizable Triene (VesiCAT)". It is based on specific UV absorbance spectral properties of a new phospholipid probe, synthesized with natural conjugated eleostearic acid extracted from Tung oil. The VesiCAT assay has been developed with two different radical generators (2,2'-azobis(2-amidinopropane) dihydrochloride; AAPH and 2,2'-azobis(2,4-dimethylvaleronitrile); AMVN), producing a constant flux of oxidant species, either in membrane or in aqueous phase. This method appears very efficient in assessing the effect of various pure antioxidant molecules in their ability to preserve liposomes from oxidative degradation. In addition, the AAPH- and AMVN-induced oxidations offer the possibility of extracting different but complementary information with respect to the antioxidants efficacy.

Lipase Activity of Tropical Oilseed Plants for Ethyl Biodiesel Synthesis and Their Typo- and Regioselectivity.

The aim of this work was to investigate lipase activities in crude extracts from Adansonia suarezensis, Adansonia grandidieri, Moringa drouhardii, Moringa oleifera, Jatropha mahafalensis, and Jatropha curcas seeds in ethanolysis and hydrolysis reactions. All crude extracts from germinated seeds showed both ethanolysis and hydrolysis activities. The influence of germination, the delipidation procedure, and the triacylglycerol/ethanol molar ratio on their ethanolysis activity was studied. Crude extracts of Jatropha and Adansonia seeds showed optimal activity at pH 8 with an optimum temperature of 30 and 40 °C, respectively. The study of the regioselectivity of crude extracts from J. mahafalensis and A. grandidieri seeds, which had the most active hydrolysis reaction, showed 1,3 regioselectivity in the hydrolysis reaction of vegetable oils. The crude extract from A. grandidieri seeds showed no typoselectivity, whereas the typoselectivity of the crude extract of J. mahafalensis seeds depended on the type of reaction.

Genetic architecture of palm oil fatty acid composition in cultivated oil palm (Elaeis guineensis Jacq.) compared to its wild relative E. oleifera (H.B.K) Cortés.

We searched for quantitative trait loci (QTL) associated with the palm oil fatty acid composition of mature fruits of the oil palm E. guineensis Jacq. in comparison with its wild relative E. oleifera (H.B.K) Cortés. The oil palm cross LM2T x DA10D between two heterozygous parents was considered in our experiment as an intraspecific representative of E. guineensis. Its QTLs were compared to QTLs published for the same traits in an interspecific Elaeis pseudo-backcross used as an indirect representative of E. oleifera. Few correlations were found in E. guineensis between pulp fatty acid proportions and yield traits, allowing for the rather independent selection of both types of traits. Sixteen QTLs affecting palm oil fatty acid proportions and iodine value were identified in oil palm. The phenotypic variation explained by the detected QTLs was low to medium in E. guineensis, ranging between 10% and 36%. The explained cumulative variation was 29% for palmitic acid C16:0 (one QTL), 68% for stearic acid C18:0 (two QTLs), 50% for oleic acid C18:1 (three QTLs), 25% for linoleic acid C18:2 (one QTL), and 40% (two QTLs) for the iodine value. Good marker co-linearity was observed between the intraspecific and interspecific Simple Sequence Repeat (SSR) linkage maps. Specific QTL regions for several traits were found in each mapping population. Our comparative QTL results in both E. guineensis and interspecific materials strongly suggest that, apart from two common QTL zones, there are two specific QTL regions with major effects, which might be one in E. guineensis, the other in E. oleifera, which are independent of each other and harbor QTLs for several traits, indicating either pleiotropic effects or linkage. Using QTL maps connected by highly transferable SSR markers, our study established a good basis to decipher in the future such hypothesis at the Elaeis genus level.

Antioxidant properties of chlorogenic acid and its alkyl esters in stripped corn oil in combination with phospholipids and/or water.

In bulk oil, it is generally thought that hydrophilic antioxidants are more active than lipophilic antioxidants. To test this hypothesis, the antioxidant activity of phenolics with increasing hydrophobicity was evaluated in stripped corn oil using both conjugated diene and hexanal measurements. Chlorogenic acid and its butyl, dodecyl, and hexadecyl esters were used as model phenolic antioxidants with various hydrophobicities. Results showed that hydrophobicity did not correlate well with antioxidant capacity. The combination of chlorogenic acid derivatives with dioleoylphosphatidylcholine (DOPC) and/or water was also studied to determine if the physical structure in the oil affected antioxidant activity. DOPC alone made hexadecyl chlorogenate a less effective antioxidant, but it did not change the antioxidant capacity of chlorogenic acid. In contrast, the combination of DOPC and water (∼400 ppm) renders chlorogenic acid a less active antioxidant, whereas it does not change the activity of hexadecyl chlorogenate. These results show, in bulk oil, that intrinsic parameters such as the hydrophobicity of lipophilized phenolics do not exert a strong influence on antioxidant capacity, but they can be highly influential if potentialized by extrinsic factors such as physical structures in the oil.

Carica papaya lipase: a naturally immobilized enzyme with interesting biochemical properties.

Triacylglycerol (TAG) lipases have been thoroughly characterized in mammals and microorganisms, whereas very little is known about plant TAG lipases. The lipolytic activity occurring in all the laticies is known to be associated with sedimentable particles, and all attempts to solubilize the lipolytic activity of Carica papaya latex have been unsuccessful so far. However, some of the biochemical properties of the lipase from Carica papaya latex (CPL) were determined from the insoluble fraction of the latex. The activity was optimum at a temperature of 37°C and a pH of 9.0, and the specific activities of CPL were found to be 2,000 ± 185 and 256 ± 8 U/g when tributyrin and olive oil were used as substrates, respectively. CPL was found to be active in the absence of any detergent, whereas many lipases require detergent to prevent the occurrence of interfacial denaturation. CPL was inactive in the presence of micellar concentrations of Triton X-100, sodium dodecyl sulfate (SDS) and tetradecyl trimethylammonium bromide (TTAB), and still showed high levels of activity in the presence of sodium taurodeoxycholate (NaTDC) and the zwitterionic Chaps detergent. The effects of various proteases on the lipolytic activity of CPL were studied, and CPL was found to be resistant to treatment with various enzymes, except in the presence of trypsin. All these properties suggest that CPL may be a good candidate for various biotechnological applications.

Conjugated autoxidizable triene (CAT) assay: a novel spectrophotometric method for determination of antioxidant capacity using triacylglycerol as ultraviolet probe.

Described here is a novel spectrophotometric method for estimating antioxidant capacity in a 96-well microplate using as UV probes the conjugated triene triacylglycerols (TAGs) naturally present in tung oil. The TAGs of this commercially available oil contain around 86% eleostearic acid, an octadecatrienoic acid with conjugated trienes exhibiting strong UV absorption at 273 nm. In an oil-in-water emulsion at 37 degrees C, the azo initiator 2,2'-azobis(2-amidinopropane) dihydrochloride generated a constant flux of peroxy radicals, which destroyed the conjugated trienes. The absorbance decay at 273 nm, related to oxidative degradation of conjugated triene TAGs, was monitored in the absence and presence of various concentrations of antioxidants. To validate this new method, the antioxidant capacity of four phenolic compounds (gallic acid, (-)-epicatechin, chlorogenic acid, and rosmarinic acid) was measured using an area-under-the-curve calculation and expressed as Trolox equivalents, the value for Trolox being taken as reference. The order of efficacy was found to be rosmarinic acid > chlorogenic acid approximately epicatechin>Trolox>gallic acid, which can be partially explained by the number of catecholic moieties and the polarity of these antioxidants. Moreover, the conjugated autoxidizable triene (CAT) assay provided good insight into antioxidant behavior (i.e., retarder or chain breaker). The same molecules were then analyzed in 2,2-diphenyl-1-picrylhydrazyl and fluorescein-based oxygen radical absorbance capacity assays, and the results are discussed critically with respect to those obtained with the CAT assay. This new method may constitute an easy-to-use, sensitive, and high-throughput in vitro protocol for evaluation of the antioxidant capacity of pure molecules and natural extracts in lipid oxidation.

Quantitative and qualitative study of gastric lipolysis in premature infants: do MCT-enriched infant formulas improve fat digestion?

Intragastric fat digestion was investigated by analyzing the products of lipolysis and the gastric lipase (HGL) levels of premature infants fed with a formula enriched with medium chain triglycerides (MCT) and those of infants fed with human milk. Infants were fed using a gastric tube and the gastric contents were aspirated twice a day for 5 d, before and at various times after gavage feeding. HGL levels were measured using the pHstat technique. After extraction, lipids were separated and quantified using thin-layer chromatography coupled to a flame ionization detector. Fatty acid methyl esters were analyzed by gas chromatography. HGL concentration increased during digestion, reaching 77.4 +/- 43.1 microg/mL (around 75% of those recorded in adults). Mean HGL output was 115 +/- 43 microg for 3 h and the overall intragastric lipolysis was 6.1 +/- 2.6%. Although the formula was enriched with octanoic and decanoic acid, the main fatty acids released in the stomach were palmitic (C16:0, 17.03 +/- 0.23% wt/wt) and oleic (C18:1 n-9, 28.23 +/- 1.26% wt/wt) acid. Similar results were obtained with infants fed with human milk. MCT supplementation has no quantitative or qualitative effects on the intragastric lipolysis, which is not higher in premature infant than in adults.

Comparison of the lipase activity in hydrolysis and acyl transfer reactions of two latex plant extracts from babaco (Vasconcellea x Heilbornii Cv.) and Plumeria rubra: Effect of the Aqueous microenvironment.

The enzymatic properties of Plumeria rubra latex have been evaluated for the first time, showing a high activity in both hydrolysis and synthesis reactions, and compared to the biocatalytic behavior of babaco (Vasconcellea x Heilbornii cv.) latex. Both biocatalysts have been optimized by studying the various parameters that influence reaction kinetics. The optimum temperatures for hydrolysis reactions were 50 and 55 degrees C for babaco and Plumeria, respectively. The optimum pH for babaco latex was 7, whereas for Plumeria latex, two optimal pH values (4 and 7) were observed. With regard to esterification and acyl transfer reactions such as alcoholysis and interesterification, the influence of thermodynamic water activity on reaction yields was determined and correlated with water sorption and desorption isotherms. When babaco latex is used as a biocatalyst, optimal synthesis reaction yields are obtained when the enzymatic extract is stabilized at a water activity value of 0.38, which corresponds to a water content of 5.7%. This optimal level of hydration is located on the linear portion of the biocatalyst's sorption isotherm, where the water molecules exhibit high-energy interactions with the protein network. In synthesis reactions (esterification, alcoholysis, and interesterification) biocatalyzed by Plumeria latex, correlation between best reaction yields and water activity cannot be done. Indeed, the sorption isotherm plot has an atypical shape, indicating that water might be trapped in the latex matrix and, consequently, that the water content of the biocatalyst is highly dependent on the hydration history of the latex.

Altered processing of the neurotensin/neuromedin N precursor in PC2 knock down mice: a biochemical and immunohistochemical study.

Neurotensin (NT) and neuromedin N (NN) are generated by endoproteolytic cleavage of a common precursor molecule, pro-NT/NN. To gain insight into the role of prohormone convertases PC1, PC2, and PC7 in this process, we investigated the maturation of pro-NT/NN in the brain of PC7 (PC7-/-), PC2 (PC2-/-), and/or PC1 (PC1+/- and PC2-/-; PC1+/-) knock down mice. Inactivation of the PC7 gene was without effect, suggesting that this convertase is not involved in the processing of pro-NT/NN. By contrast, there was a 15% decrease in NT and a 50% decrease in NN levels, as measured by radioimmunoassay, in whole brain extracts from PC2 null as compared with wild type mice. Using immunohistochemistry, we found that this decrease in pro-NT/NN maturation products was uneven and that it was most pronounced in the medial preoptic area, lateral hypothalamus, and paraventricular hypothalamic nuclei. These results suggest that PC2 plays a critical role in the processing of pro-NT/NN in mouse brain and that its deficiency may be compensated to a regionally variable extent by other convertases. Previous data have suggested that PC1 might be subserving this role. However, there was no change in the maturation of pro-NT/NN in the brain of mice in which the PC1 gene had been partially inactivated, implying that complete PC1 knock down may be required for loss of function.