RP-HPLC/MS-APCI Analysis of Branched Chain TAG Prepared by Precursor-Directed Biosynthesis with <Emphasis Type="Italic">Rhodococcus</Emphasis><Emphasis Type="Italic">erythropolis</Emphasis>

Reversed phase liquid chromatography–atmospheric pressure chemical ionization mass spectrometry (RP-HPLC/MS-APCI) was used to analyze both synthetic triacylglycerols (TAG) having 1–3 branched fatty acids (FA) in the molecule, and natural TAG prepared by precursor directed biosynthesis from valine, leucine and isoleucine and the corresponding branched short-chain acids in cultivations of Rhodococcus erythropolis. The technique made it possible to identify and quantify TAG differing in a single branched-chain FA. Altogether 11 TAG were synthesized, out of which 8 were synthesized stereospecifically. Branched- and straight-chain-TAG were separated and identified while TAG differing only in iso or anteiso FA could not be separated. The APCI mass spectra of iso-, anteiso- and straight-chain TAG were completely identical. The natural material was found to contain 19 TAG having at least one branched FA. Cultivation on six different substrates showed, apart from the presumed and common incorporation of precursors to iso-even, iso-odd and anteiso FA, also some unusual features such as an increase in the content of odd-FA after the addition of Val (attributed to catabolism of Val to propionate) or the appearance of branched monounsaturated FA. The two-sample paired t test, when applied to the TAG, showed that only the pair Val and isobutyrate differ in incorporation into FA—see, e.g. proportions of M/M/O and brM/brM/O (1.2:1.2 and 1.9:1.2, respectively). Also, incorporation of Val (isobutyrate) yielded only TAG having two branched FA in the molecule, whereas Leu and Ile (isovalerate and 2-methyl-butyrate) gave only TAG with a single branched FA in the molecule.     

Ascorbic Acid Supplementation Improves Skeletal Muscle Growth in Pacu (Piaractus mesopotamicus) Juveniles: In Vivo and In Vitro Studies

In fish, fasting leads to loss of muscle mass. This condition triggers oxidative stress, and therefore, antioxidants can be an alternative to muscle recovery. We investigated the effects of antioxidant ascorbic acid (AA) on the morphology, antioxidant enzyme activity, and gene expression in the skeletal muscle of pacu (Piaractus mesopotamicus) following fasting, using in vitro and in vivo strategies. Isolated muscle cells of the pacu were subjected to 72 h of nutrient restriction, followed by 24 h of incubation with nutrients or nutrients and AA (200 µM). Fish were fasted for 15 days, followed by 6 h and 15 and 30 days of refeeding with 100, 200, and 400 mg/kg of AA supplementation. AA addition increased cell diameter and the expression of anabolic and cell proliferation genes in vitro. In vivo, 400 mg/kg of AA increased anabolic and proliferative genes expression at 6 h of refeeding, the fiber diameter and the expression of genes related to cell proliferation at 15 days, and the expression of catabolic and oxidative metabolism genes at 30 days. Catalase activity remained low in the higher supplementation group. In conclusion, AA directly affected the isolated muscle cells, and the higher AA supplementation positively influenced muscle growth after fasting.     

Mechanism Controlling High-Temperature Degradation of Sunflower Oil Triacylglycerols in the Absence of Oxygen

The aim of the present study is to describe the mechanism controlling heat-induced formation of sunflower oil triacylglycerol and fatty acid methyl ester oligomers. The unique combination of high-performance size-exclusion chromatography with hyphenated electrospray ionization mass spectrometry (MS), atmospheric pressure chemical ionization-MS, and high-temperature gas chromatography-MS techniques allows differentiating between radical coupling species and Diels–Alder cycloadducts. Targeted analysis of thermally degraded sunflower oils confirms the exact structures of various acyclic oligomers accompanied by less-abundant products of pericyclic transformations. A series of model experiments simulate the impact of dienophile nature on the course of Diels–Alder reactions. Thus, α-tocopherylquinone, δ-tocopherylquinone, and methyl-(E)-11-oxoundec-9-enoate are synthesized as naturally occurring dienophiles bearing electron-withdrawing groups. The geometry of poor dienophiles does not affect concerted cyclization, while the structure of electron deficient dienophiles can overcome low reactivity. Practical Application: In the absence of oxygen, heat-induced degradation of polyunsaturated triacylglycerols proceed predominantly via a radical pathway, whereas concerted reactions represent minor mechanisms. Sunflower oil triacylglycerol molecules in the system without propagation stage can be effectively protected by natural and/or synthetic antioxidants. Application of chelates is also recommended. However, antioxidant-derived quinones, such as α-tocopherylquinone, can enter the Diels–Alder reaction even more easily than dienophiles without electron-withdrawing groups. Unsaturated core aldehydes possess the same reactivity. Examination of the mechanism controlling high-temperature degradation of triacylglycerols is especially important for processing engineers in edible oil refineries and food technologists. New perspective may help them to minimize undesirable changes in polyunsaturated species.     

Application of a hop by‐product in brewing: reduction in the level of haze‐active prolamines and improved antioxidant properties of the beer

After supercritical CO₂ extraction of hops or formation of lupulin‐enriched pellets (type 45), polyphenol‐rich by‐products remain. These have some industrial and agricultural applications (preparation of polyphenol extracts and in the production of low‐bitterness beers). In this work, both materials were used in pilot‐scale brewing experiments for the production of beer enriched with polyphenols. Samples of beer made from these residues underwent analyses for haze‐active prolamines, total polyphenols, phenolic monomers, antioxidant capacity, bitterness and colour. An almost identical decrease in the content (from hopped wort to final beer) of the prolamines of approximately 30% was detected in standard beer as well as in the beer prepared from the hop residue (HR 1) after supercritical CO₂ extraction. In contrast, the level of prolamines was reduced by more than 50% in the beer following addition of the hop residue (HR 2) after production of lupulin enriched pellets. Both hop materials contributed to an increase in the antioxidant capacity of the final beer by 21.5 and 11.9%, respectively. Another part of this work was aimed at the investigation of colour and bitterness changes in beers made ??using hop by‐products. Addition of the material HR 2 into the wort caused an increase in colour and in bitterness of the matured beer by 20 and 11% rel., respectively. Copyright © 2014 The Institute of Brewing & Distilling     

Development of a LC-IT-TOF MS Procedure to Quantify Veterinary Drug Residues in Milk Employing a QuEChERS Approach

A sensitive liquid chromatography coupled to high-resolution tandem mass spectrometry (LC-IT-TOF MS) with electrospray ionization method for the identification and quantification of five antibiotic residues in milk has been developed. For sample preparation, a simple modification of the QuEChERS method was used with further sample cleanup using dispersive solid phase extraction according to AOAC official standards 2007.01. The method yielded acceptable accuracy values for each drug at each level, with mean recoveries (n?=?3) ranging from 83 to 92 %. The recovery of antibiotics of the intra-assay ranged from 85 to 95 % (RSD?<?9 %) and the inter-assay from 84 to 95 % (RSD?<?11 %). A total of 31 milk samples, either pasteurized or fresh whole milk, were analyzed following the method. Amoxicillin and oxytetracycline (two of the pasteurized milk samples), ampicillin and amoxicillin (3 of 25 fresh milk samples), and tetracycline and oxytetracycline (two other fresh milk samples out of 25) were detected at levels below the current Brazilian maximum residue levels.     

Stability of vitamin A during storage of enteral feeding formulas

Vitamin A (13-cis-retinol, all-trans-retinol and total carotenes) content was evaluated in 15 samples of enteral feeding formulas during 3–6, 9 and 12 months of storage in closed containers, protected from light and at room temperature (22–30 °C). All formulas were nutritionally complete and/or recommended for different diseases. Equivalent batches were also evaluated after preparation and refrigerated storage for 24 h. Nutrient content did not decrease during both storage scenarios. Stability of vitamin A in the enteral feeding formulas, as determined in this study, is probably due to production and storage conditions, e.g. absence of oxygen in contact with the packaged product, the technological process used in the microencapsulation of vitamins, and protected storage in the absence of light and at a temperature maximum of 30 °C.     

The effect of lactose, NaCl and an aero/anaerobic environment on the tyrosine decarboxylase activity of Lactococcus lactis subsp. cremoris and Lactococcus lactis subsp. lactis

The aim of this work was to study, under model conditions, combined effects of the concentration of lactose (0-1% w/v), NaCl (0-2% w/v) and aero/anaerobiosis on the growth and tyramine production in 3 strains of Lactococcus lactis subsp. lactis and 2 strains of L. lactis subsp. cremoris. The levels of the factors tested were chosen with respect to the conditions which can occur during the real process of natural cheese production, including the culture temperature (10 ± 1 °C). In all strains tested, tyrosine decarboxylation was most influenced by NaCl concentration; the highest production of tyramine was obtained within the culture with the highest (2% w/v) salt concentration applied. Two of the strains L. lactis subsp. lactis produced tyramine only in broth with the highest NaCl concentration tested. In the remaining 3 strains of L. lactis, tyramine was detected under all conditions applied. The tested concentration of lactose and aero/anaerobiosis had a less significant effect on tyramine decarboxylation. However, it was also found that at the same concentrations of NaCl and lactose, a higher amount of tyramine was detected under anaerobic conditions. In all strains tested, tyramine decarboxylation started during the active growth phase of the cells.     

Is contextual animation needed in multimedia learning games for children? An eye tracker study

The present study investigates affective-motivational, attention, and learning effects of unexplored emotional design manipulation: Contextual animation (animation of contextual elements) in multimedia learning game (MLGs) for children. Participants (N = 134; M_age = 9.25; Grades 3 and 4) learned either from an experimental version of the MLG with a high amount of contextual animation or from an identical MLG with no contextual animation (control). Children strongly preferred (χ² = 87.04, p < .001) and found the experimental version more attractive (p < .001, d = −1.11). No significant differences in overall enjoyment and learning outcomes were found. Attention differences, measured by dwell times and fixation durations, were small and reached only borderline significance (p = .035; d = −0.39). The implication is that contextual animation in MLG for children increases such instructional materials' attractiveness without compromising cognitive processes needed for learning; however, it does not lead to their higher instructional efficiency.     

To solve or to observe? The case of problem-solving interactivity within child learning games

We explored whether problem-solving interactivity within an instructional game fosters learning for children aged 8–10 years. Participants (N = 139) studied a biological topic either through a game-based learning environment (in which they solved assigned problems by interacting with a plant model) or from a standard learning environment (in which they observed how the problems were solved in an animation) (i.e., between-subject design). The treatments were equal with regard to learning content and guidance. No between-group differences in learning outcomes were detected (comprehension: d = 0.16; transfer: d = −0.01). Self-rating of enjoyment tended to be higher in the game group (d = 0.32), and when the children could choose between the treatments in a free-choice period, they strongly preferred the interactive game (87.5%). The results suggest that both the interactive and the non-interactive treatments are useful, but their applicability may differ depending on the learning context (e.g., school vs. home).