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Vegetables rich in bitter-tasting phytochemicals may exert enhanced beneficial effects against key factors associated with type two diabetes (T2D). This study investigates whether selected cultivars of bitter and strong-tasting (BST) Brassica and root vegetables exert greater health benefits on T2D patients compared to equivalent modern mild and sweet tasting (MST) vegetables. A 12-week randomized, controlled, parallel intervention study involved 92 T2D patients, who were allocated three different diets: (1) 500 g daily of bitter and strong-tasting (BST) vegetables, (2) 500 g daily of mild and sweet-tasting (MST) vegetables, (3) 120 g daily MST normal diet (control). Both vegetable diets contained root vegetables and cabbages selected based on sensory differences and content of phytochemicals. Prior to and after the study, all participants underwent an oral glucose tolerance test (OGTT), 24 h blood pressure measurements, DEXA scans, and fasted blood samples. Both diets high in vegetables significantly reduced the participants’ BMI, total body fat mass, and HbA1c levels compared to control, but in the BST group, significant differences were also found regarding incremental area under the curve glucose 240 min (OGTT) and fasting glucose levels. A high daily intake of root vegetables and cabbages showed significant health improvements in both vegetable groups. BST vegetables had the greatest impact on insulin sensitivity, body fat mass, and blood pressure compared to control, moreover, they further improved glycemic control compared to MST vegetables.

The spectral qualities of light contain environmental information used by plants as cues to modify their biology in order to adapt and survive. LED based lamps, with their highly definable spectral properties, thus hold great perspectives for use in greenhouse production. The effects of the spectral qualities of light on plant physiology is still not fully understood, and a great fraction of the current knowledge further comes from studies looking at short term effects or using monochromatic light treatments in enclosed growth chambers, thus far from the light conditions of greenhouse production. The objective of this study was to determine whether spectral modifications to the supplemental LED lighting in greenhouse production could modify post-cultivation performance of basil, which is prone to chilling injuries (CI). We cultivated basil under greenhouse conditions using four different supplemental LED light treatments; 80%Red/20%Blue, 80%Red/20%Blue + UV-A, 40%Red/60%Blue and 80%Red/20%Green, followed by a post-cultivation shelf life simulation including a chilling treatment. Stomatal anatomical features and levels of hormonal regulators, abscisic acid (ABA) and ABA glucosylester (ABA-GE), along with soluble sugars, fatty acids and lipoxygenase activity (LOX) were determined at the end of the growing period. Without compromising grow rates, the light treatments showed marked effects on the chilling tolerance and shelf life performance with increasing ratios of blue light having negative effects, and green light having positive effects on chilling tolerance. Differences in shelf life performance seemed mediated solely by factors related to stomatal functioning. Stomatal density (number of stomata per leaf area, SD) increased with increasing ratios of blue light whereas green light showed indications to decrease SD. [ABA] and [ABA-GE] showed positive correlation to SD, whereas the ratio of [ABA-GE]/[ABA] conversely correlated negatively with increased SD. Our results thus suggest an important relationship between leaf water retention and post-cultivation chiling tolerance in basil.

Scope Proteins constitute an important part of the human diet, but understanding of the effects of different dietary protein sources on human metabolism is sparse. We aimed to elucidate diet-induced metabolic changes through untargeted urinary metabolomics after four weeks of intervention with lean-seafood or nonseafood diets. It is shown that lean-seafood intake reduces urinary excretion of metabolites involved in mitochondrial lipid and energy metabolism possibly facilitating a higher lipid catabolism in healthy subjects. Methods In a randomized controlled trial with crossover design, 20 healthy subjects consumed two balanced diets that varied in main protein sources for 4 weeks. Morning spot urine samples were collected before and after each intervention period. Untargeted metabolomics based on (1) H NMR spectroscopy and LC-MS analyses were applied to characterize the urinary metabolic response to the interventions. Results The lean-seafood diet period reduced the urinary level of l-carnitine, 2,6-dimethylheptanoylcarnitine, and N-methyl-2-pyridone-5-carboxamide, relative to the nonseafood period. The dietary analysis revealed that the higher urinary level of trimethylamine-N-oxide after the lean-seafood diet period and guanidinoacetate and 3-methylhistidine after the nonseafood diet period was related to the endogenous content of these compounds in the diets. Conclusions Our data reveal that 4 weeks of lean-seafood intake reduces urinary excretion of metabolites involved in mitochondrial lipid and energy metabolism possibly facilitating a higher lipid catabolism in healthy subjects after the lean-seafood intake.

Storage-induced changes are known to be more prominent in lactose-hydrolyzed (LH) milk compared to conventional milk. Therefore the present study aimed at identifying off-flavors resembling from formation of volatiles during storage of ultra-high temperature treated (UHT) LH milk and conventional UHT milk. Further, the influence of heat processing, indirect or direct, on UHT LH milk was also examined. Storage-induced changes in sensory attributes, volatiles and primary amines were investigated during a 4 months period. Conventional UHT milk (with 5 % lactose) processed using indirect heat treatment (CONVI) and two types of UHT LH milk (with less than 0.01 % lactose) produced using either direct heat treatment (LHD) or indirect heat treatment (LHI) were represented in the study. Sensory descriptive analysis showed that fresh samples of CONVI, LHI and LHD differed in sensory properties and the samples could be differentiated according to boiled and stale aroma as well as color saturation. Differentiation of the fresh samples based on the volatile gas chromatography–mass spectrometry profile was not achievable. During storage, samples developed differently with respect to sensory characteristics, volatiles and the amount of primary amines. Partial least squares models (PLS1) including only methyl ketones and aldehydes showed that 2-butanone, 2-pentanone, 2-heptanone, 2-nonanone, heptanal, octanal and nonanal predicted stale flavor. Bitter taste, on the other hand, correlated with the amount of primary amines (Pearson’s correlation, r 2 = 0.71). This finding indicates that storage-induced changes in sensory characteristics, volatiles and primary amines depend on both differences in lactose content and the applied heat processing.

Proteolytic activity in milk may release bitter-tasting peptides and generate free amino terminals that react with carbohydrates, which initiate Maillard reaction. Ultrahigh temperature (UHT) heat treatment inactivates the majority of proteolytic enzymes in milk. In lactose-hydrolyzed milk a β-galactosidase preparation is applied to the milk after heat treatment, which has proteolytic side activities that may induce quality deterioration of long-term-stored milk. In the present study proteolysis, glycation, and volatile compound formation were investigated in conventional (100% lactose), filtered (60% lactose), and lactose-hydrolyzed (

Genetic variation is the most influential factor for carrot (Daucus carota L.) composition. However, difference in metabolite content between carrot varieties has not been described by NMR, although primary metabolites are important for human health and sensory properties. The aim of the present study was to investigate the effect of genotype on carrot metabolite composition using a (1)H NMR-based metabolomics approach. After extraction using aqueous and organic solvents, 25 hydrophilic metabolites, β-carotene, sterols, triacylglycerols, and phospholipids were detected. Multiblock PCA showed that three principal components could be identified for classification of the five carrot varieties using different spectroscopic regions and the results of the two solvent extraction methods as blocks. The varieties were characterized by differences in carbohydrate, amino acid, nucleotide, fatty acid, sterol, and β-carotene contents. (1)H NMR spectroscopy coupled with multiblock data analysis was an efficient and useful tool to map the carrot metabolome and identify genetic differences between varieties.

Recently we showed that exchanging intact casein with extensively hydrolysed casein in Western diets prevented diet-induced obesity in obesity-prone C57BL/6J mice. To gain further insight into the underlying mechanisms for the metabolic alterations induced by intake of hydrolysed casein, we performed an exploratory investigation using proton NMR spectroscopy, multi-block PCA (MBPCA) and a multi-compartment model including analyses of plasma, urine, faeces and tissue samples from mice fed diets with intact or hydrolysed casein and 16 or 32 energy% protein. The MBPCA superscores showed a clear separation between samples from mice fed intact and hydrolysed casein diets, respectively. Block loadings revealed that fecal fat content was higher, and tissue and plasma lipid levels were lower in mice fed hydrolysed casein diets compared with mice fed intact casein. Amino acid metabolism was also altered by dietary protein form, and levels of branched-chain amino acids were higher in faeces and urine and lower in plasma and spleen in mice fed hydrolysed protein. Moreover, hepatic levels of the sulphur-containing metabolites taurine and glutathione were increased in mice fed hydrolysed casein, and hepatic glycogen amount was increased in mice fed hydrolysed casein. In contrast, the levels of glucose and its metabolite lactate were reduced in faeces, liver and plasma. Taken together, NMR-based metabolomic analyses indicated that pathways within lipid, amino acid and carbohydrate metabolism were altered by intake of hydrolysed casein, and that these alterations are likely to be underlying mechanisms for the observed prevention against diet-induced obesity associated with hydrolysed casein intake.

Metabolic changes were investigated in overwintering Jerusalem artichoke (Helianthus tuberosus L.) tubers using proton nuclear magnetic resonance ((1)H NMR) metabolomics. Three varieties were studied; as a result of overwintering, the amount of inulin was found to decrease in Jerusalem artichoke tubers. This was mainly due to its conversion to sucrose and, at the same time, formation of inulin with a lower degree of polymerization. Major effects on the concentration of citric acid, malic acid, γ-aminobutyric acid (GABA), and adenosine were also found. Intriguingly, malic acid concentration increased and citric acid concentration decreased. These changes, together with an increase in sucrose and GABA concentrations, were ascribed to mobilization of nutrients prior to sprouting, suggesting that malic acid and GABA serve as carbon and nitrogen sources during sprouting of Jerusalem artichokes.

Thirty four varieties of sour cherries (Prunus cerasus) were investigated for their total antioxidant activity, Caco-2 cancer cell proliferation inhibitory activity and effect on prostaglandin E2 (PGE2) production. Total phenolic content, oxygen radical absorbance capacity (ORAC) and cancer cell proliferation inhibitory activity of sour cherries were closely correlated but not PGE2 production. The cultivars 'Birgitte × Bottermo', 'Fanal' and 'Tiki' were the three cultivars with the highest ORAC values (180, 147 and 133 µmol TE/g, respectively) and inhibition against Caco-2 cancer cell proliferation (74%, 79% and 73%, respectively). 'Stevnsbaer Birgitte' (22%) and 'Stevnsbaer Viki' (22%) inhibited PGE2 production with a similar potency as the positive controls indomethacin and NS-398. Significant differences between cultivars in all bioactivity experiments indicated that selection of cultivars is important to obtain sour cherries with better potential health promoting effects.

The aim of the present study was to investigate the relationship between the metabolite profile of milk and important technological properties by using nuclear magnetic resonance (NMR)-based metabolomics. The metabolomics approach was introduced for the metabolic profiling of a set of milk samples from two dairy breeds representing a wide span in coagulation properties. The milk metabolite profiles obtained by proton and carbon NMR spectroscopy could be correlated to breed and, more interestingly, also with the coagulation profile, as established by traditional methods by using principal component analysis (PCA). The metabolites responsible for the separation into breed could mainly be ascribed to carnitine and lactose, whereas the metabolites varying in the samples with respect to coagulation properties included citrate, choline, carnitine, and lactose. The results found in the present study demonstrated a promising potential of NMR-based metabolomics for a rapid analysis and classification of milk samples, both of which are useful for the dairy industry.

Characterization of peroxides by size exclusion chromatography (SEC) of milk following exposure to riboflavin and light showed that hydrogen peroxide was the most abundant peroxide formed since it could be removed by catalase. Formation of peroxides after separation by SEC showed that hydrogen peroxide formation was primarily increased in the presence of caseins and ascorbate, although whey proteins also were found to contribute. Caseins and beta-lactoglobulin also formed catalase-resistant peroxides, presumably protein hydroperoxides. A catalase-resistant and unstable peroxide was observed in fractions containing urate. Experiments performed with pure urate suggested that urate radicals reacted further with superoxide leading to a urate hydroperoxide. Electron paramagnetic resonance spectroscopy using spin-traps showed that the presence of oxygen was required for urate radical formation, which could be assigned as nitrogen-centered radicals. These results suggest a new route during light-induced oxidation sensitized by flavins, in effect making urate pro-oxidative.

Radical scavenging activities of bovine milk components were quantified following size exclusion chromatography (SEC) with postcolumn characterization of fractions using the scavenging of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radicals (ABTS*(+)) in the Trolox equivalent antioxidant capacity (TEAC) assay and peroxyl radicals formed from cleavage of 2,2'-azobis(2-amidinopropane) (AAPH) in the oxygen radical absorbance capacity (ORAC) fluorometric assay. Caseins were quantitatively the major radical scavenger species in both assays, whereas beta-lactoglobulin (beta-lg) and alpha-lactalbumin (alpha-la) were much less active and only in the peroxyl radical assay. The radical scavenging activity of the caseins could be quantitatively accounted for by their constituent amino acids, as there were no effects of denaturing agents or complete digestion with proteases. In contrast, the activities of the whey proteins were dependent on denaturation or partial hydrolysis and dominated by the free thiol in beta-lg. A component in milk serum with a molecular mass of approximately 100 kDa contributed significantly to both ABTS*(+) and peroxyl radical scavenging but was absent in whey. This radical scavenger was identified as beta-casein. The only significant low molecular weight radical scavenger species were identified as ascorbate and urate in both assays.

Dairy products are an important component in the Western diet and represent a valuable source of nutrients for humans. However, a reliable dairy intake assessment in nutrition research is crucial to correctly elucidate the link between dairy intake and human health. Metabolomics is considered a potential tool for assessment of dietary intake instead of traditional methods, such as food frequency questionnaires, food records, and 24-h recalls. Metabolomics has been successfully applied to discriminate between consumption of different dairy products under different experimental conditions. Moreover, potential metabolites related to dairy intake were identified, although these metabolites need to be further validated in other intervention studies before they can be used as valid biomarkers of dairy consumption. Therefore, this review provides an overview of metabolomics for assessment of dairy intake in order to better clarify the role of dairy products in human nutrition and health.

Jerusalem artichoke (Helianthus tuberosus L.) tubers have many health benefits and are suitable for human consumption. The tubers store energy as the fructose polymer inulin. The content of inulin and the degree of polymerization is dependent on cultivar, growing conditions, harvest time, and the maturity of the tubers. Jerusalem artichoke tubers can be stored in the field during winter, or in cold rooms. During storage inulin is degraded, and shorter chain inulin polymers and sucrose are formed. The content of phenolic compounds in Jerusalem artichoke tubers is highest in mature tubers and declines during winter. The major phenolic compounds are the phenolic acids chlorogenic acid, gallic acid, salicylic acid, and caffeic acid. The content of phenolic compounds is 2–12 times higher in the peel than in the flesh of the tubers.

Milk contains lipids, carbohydrates, proteins, immunoglobulins, peptides, amino acids, nucleotides, vitamins, minerals, and other metabolites. Many factors influence the milk composition; feed, seasonal variation, genetics, health, and number of and stage of lactation. There is a clear correlation between the milk composition and its nutritional value, technological properties, and content of functional ingredients. For a number of years, breeding programs have been in place to actively improve milk quality. Additionally, a large focus has been on the effect of feed on milk quality. However less attention has been paid to changes in the milk metabolite profile through alterations in feeding regime. In bovine milk, NMR-based metabolomics has previously been used to identify small molecules in several studies. The use of NMR metabolomics to assess the milk metabolome and in dairy research in general has recently been reviewed. In the present study NMR spectroscopy was used to identify how changes in the feeding regime could cause changes in the milk metabolome. More specifically, we wanted to examine if addition of oregano (Origanum vulgare) and caraway (Carum carvi) essential oils in the feed, which are known to introduce differences in the milk volatile compounds immediately upon addition, would also affect the milk metabolite profile.

The amount and form of dietary casein have been shown to affect energy metabolism and lipid accumulation in mice, but the underlying mechanisms are not fully understood. We investigated 48 hrs urinary metabolome, hepatic lipid composition and gene expression in male C57BL/6J mice fed Western diets with 16 or 32 energy% protein in the form of extensively hydrolyzed or intact casein. LC-MS based metabolomics revealed a very strong impact of casein form on the urinary metabolome. Evaluation of the discriminatory metabolites using tandem mass spectrometry indicated that intake of extensively hydrolyzed casein modulated Phase II metabolism associated with an elevated urinary excretion of glucuronic acid- and sulphate conjugated molecules, whereas glycine conjugated molecules were more abundant in urine from mice fed the intact casein diets. Despite the differences in the urinary metabolome, we observed no differences in hepatic expression of genes involved in Phase II metabolism, but it was observed that expression of Abcc3 encoding ATP binding cassette c3 (transporter of glucuronic acid conjugates) was increased in livers of mice fed hydrolyzed casein. As glucuronic acid is derived from glucose and sulphate is derived from cysteine, our metabolomic data provided evidence for changes in carbohydrate and amino acid metabolism and we propose that this modulation of metabolism was associated with the reduced glucose and lipid levels observed in mice fed the extensively hydrolyzed casein diets.

We describe a time-saving protocol for the processing of LC-MS-based metabolomics data by optimizing parameter settings in XCMS and threshold settings for removing noisy and low-intensity peaks using design of experiment (DoE) approaches including Plackett-Burman design (PBD) for screening and central composite design (CCD) for optimization. A reliability index, which is based on evaluation of the linear response to a dilution series, was used as a parameter for the assessment of data quality. After identifying the significant parameters in the XCMS software by PBD, CCD was applied to determine their values by maximizing the reliability and group indexes. Optimal settings by DoE resulted in improvements of 19.4% and 54.7% in the reliability index for a standard mixture and human urine, respectively, as compared with the default setting, and a total of 38 h was required to complete the optimization. Moreover, threshold settings were optimized by using CCD for further improvement. The approach combining optimal parameter setting and the threshold method improved the reliability index about 9.5 times for a standards mixture and 14.5 times for human urine data, which required a total of 41 h. Validation results also showed improvements in the reliability index of about 5-7 times even for urine samples from different subjects. It is concluded that the proposed methodology can be used as a time-saving approach for improving the processing of LC-MS-based metabolomics data.

We investigated the effect of a 24-week energy-restricted intervention with low or high dairy intake (LD or HD) on the metabolic profiles of urine, blood and feces in overweight/obese women by NMR spectroscopy combined with ANOVA-simultaneous component analysis (ASCA). A significant effect of dairy intake was found on the urine metabolome. HD intake increased urinary citrate, creatinine and urea excretion, and decreased urinary excretion of trimethylamine-N-oxide (TMAO) and hippurate relative to the LD intake, suggesting that HD intake was associated with alterations in protein catabolism, energy metabolism and gut microbial activity. In addition, a significant time effect on the blood metabolome was attributed to a decrease in blood lipid and lipoprotein levels due to the energy restriction. For the fecal metabolome, a trend for a diet effect was found and a series of metabolites, such as acetate, butyrate, propionate, malonate, cholesterol and glycerol tended to be affected. Overall, even though these effects were not accompanied by a higher weight loss, the present metabolomics data reveal that a high dairy intake is associated with endogenous metabolic effects and effects on gut microbial activity that potentially impact body weight regulation and health. Moreover, ASCA has a great potential for exploring the effect of intervention factors and identifying altered metabolites in a multi-factorial metabolomic study.

Untargeted liquid chromatography-mass spectrometry (LC-MS)-based metabolomics studies are usually carried out in both positive and negative ion modes; however, it is frequently ignored that the optimal conditions in positive ion mode and negative ion mode are often not the same. We carried out a systematic investigation on urine samples to evaluate the additive effects in negative ion mode. It was found that the widely used conditions, 0.1% formic acid (FA) and NH(4)Ac at different pH, are far from the optimum for untargeted urine metabolomics studies. Compared to 0.1% FA, the use of 1 mM acetic acid (HAc) resulted in almost three times as many detected peaks (401 vs 148) and around five times the size of the peak area (33.55 × 10(6) vs 6.47 × 10(6)). The remarkable improvement can be explained by two factors: (i) a significantly enhanced ionization efficiency due to the combination of an appropriate pH at around 4.0-4.5, the reducibility of H(+), and the high gas-phase basicity of Ac(-) and (ii) a reproducible LC separation due to an acceptable buffering capacity. Our study revealed the importance and necessity of additive optimization, which can be of benefit in related metabolomics studies.

Thirty four varieties of sour cherries (Prunus cerasus) were investigated for their total antioxidant activity, Caco-2 cancer cell proliferation inhibitory activity and effect on prostaglandin E2 (PGE2) production. Total phenolic content, oxygen radical absorbance capacity (ORAC) and cancer cell proliferation inhibitory activity of sour cherries were closely correlated but not PGE2 production. The cultivars 'Birgitte × Böttermö', 'Fanal' and 'Tiki' were the three cultivars with the highest ORAC values (180, 147 and 133 µmol TE/g, respectively) and inhibition against Caco-2 cancer cell proliferation (74%, 79% and 73%, respectively). 'Stevnsbaer Birgitte' (22%) and 'Stevnsbaer Viki' (22%) inhibited PGE2 production with a similar potency as the positive controls indomethacin and NS-398. Significant differences between cultivars in all bioactivity experiments indicated that selection of cultivars is important to obtain sour cherries with better potential health promoting effects.

Eleven blackcurrant cultivars grown with pesticide (PT) and without pesticide treatment (PF) were evaluated to compare the differences in plant growth and physical condition, total anthocyanin content, ascorbic acid content, total antioxidant capacity, effect on prostaglandin E2 (PGE2) production and anticancer cell proliferation activities. Results showed that the yield and growth of PT blackcurrants were higher. However, PF blackcurrants contained a higher amount of ascorbic acid, and displayed an increased inhibition against cancer cells compared to PT blackcurrants, indicating that PF blackcurrants have an increased potential to deliver health-promoting benefit for consumers. Significant differences were observed between blackcurrant cultivars in relation to plant growth and physical condition, total anthocyanin content and PGE2 assay, highlighting the importance of cultivar selection.