Strong and Bitter Vegetables from Traditional Cultivars and Cropping Methods Improve the Health Status of Type 2 Diabetics: A Randomized Control Trial.

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.

Effect of long-term heat exposure on rheological and intrinsic water characteristics of bone-derived beef stocks.

Bone-derived protein stocks are used in food industry to enhance taste of soups, sauces, and a range of other products. Both during commercial manufacturing and when used for culinary purposes, the stocks may be exposed to high temperatures for an extended time period. The present study investigated the effect of retention at 90°C for 0, 3, 6, 9, 24, 48, 72, and 168 hr on the functional attributes of concentrated bone-derived beef stocks (57% Dry matter (DM)). Visual inspection and rheological analyses showed that during increasing heat exposure, the gel strength as well as viscosity of the concentrated stocks decreased incrementally and significantly (P > 0.001). Nuclear magnetic resonance (NMR) relaxation measurements conducted on the beef stocks also revealed strong effects of heat exposure on the transverse (T2 ) relaxation time, which increased incrementally and significantly (P > 0.001) with longer heat exposure. Thus, the present study demonstrated that heat-induced changes in rheological properties of bone-derived beef stocks can be ascribed to changes in intrinsic water-protein interactions and water attributes as a result of heat-induced protein modifications. In conclusion, the study proves that NMR relaxometry is a valuable tool for monitoring changes in intrinsic water mobility that are manifested in modified functional attributes of concentrated beef stocks.

Consumption of Whey in Combination with Dairy Medium-Chain Fatty Acids (MCFAs) may Reduce Lipid Storage due to Urinary Loss of Tricarboxylic Acid Cycle Intermediates and Increased Rates of MCFAs Oxidation.

SCOPE: The aim of the paper is to investigate whether changes in the metabolome could explain observed changes in body composition in overweight adults after consumption of butter with high level of medium-chain fatty acids (MCFAs) in combination with casein or whey. METHODS AND RESULTS: With GC-TOF and LC-Q/MS, metabolites in plasma and urine from a 12-week randomized double-blinded human intervention including 52-abdominally overweight adults were analyzed. The participants consumed 63 g per day of milk fat (high or low in MCFAs) and 60 g per day of protein (whey or casein). Urinary loss of the tricarboxylic acid cycle metabolites and a concomitantly increase of glycerol in blood were observed in the whey + high-MCFAs group, indicating potential lower anabolic processes, such as lipogenesis, by draining substrates. High intake of MCFAs resulted in elevated level of urinary adipic (independently of protein type) and plasma sebacic acid (with whey), indicating a potential increase in oxidation of MCFAs, which might lead to energy loss. CONCLUSION: The type of protein showed highest effect on the overall metabolic profiles, but ω-oxidation of MCFAs in the liver seemed to be the main reason for the observed reduction in body fat mass after consumption of high MCFAs, independent of type of protein.

Lean-seafood intake decreases urinary markers of mitochondrial lipid and energy metabolism in healthy subjects: Metabolomics results from a randomized crossover intervention study.

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.

Intake of hydrolyzed casein is associated with reduced body fat accretion and enhanced phase II metabolism in obesity prone C57BL/6J mice.

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.

Mapping the variation of the carrot metabolome using 1H NMR spectroscopy and consensus PCA.

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, ß-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 ß-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.

Photoperiodic variations induce shifts in the leaf metabolic profile of Chrysanthemum morifolium.

Plants have a high ability to adjust their metabolism, growth and development to changes in the light environment and to photoperiodic variation, but the current knowledge on how changes in metabolite contents are associated with growth and development is limited. We investigated the effect of three different photoperiodic treatments with similar daily light integral (DLI) on the growth responses and diurnal patterns in detected leaf metabolites in the short day plant Chrysanthemum×morifolium Ramat. Treatments were long day (LD, 18h light/6h dark), short day (SD, 12h light/12h dark) and short day with irregular night interruptions (NI-SD,12h light/12h dark, applied in a weekly pattern, shifting from day-to-day). Photoperiodic variation resulted in changes in the phenotypic development of the plants. The plants grown in the SD treatment started to initiate reproductive development of the meristems and a decrease in leaf expansion resulted in lower leaf area of expanding leaves. In contrast, plants in the NI-SD and LD treatments did not show reproductive development at any stage and final leaf area of the expanding leaves was intermediate for the NI-SD plants and largest for the LD plants. Photoperiodic variation also resulted in changes in the leaf metabolic profile for most of the analysed metabolites, but only carbohydrates, citrate and some amino acids displayed a shift in their diurnal pattern. Further, our results illustrated that short days (SD) increased the diurnal turnover of 1-kestose after 2 weeks, and decreased the overall contents of leaf hexoses after 3 weeks. In the two other treatments a diurnal turnover of 1-kestose was not stimulated before after 3 weeks, and hexoses together with the hexose:sucrose ratio steadily increased during the experiment. Our results enlighten the plasticity of leaf growth and metabolism to environmental changes, and demonstrate that diurnally regulated metabolites not always respond to photoperiodic variation.

Time-saving design of experiment protocol for optimization of LC-MS data processing in metabolomic approaches.

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.

Enhancing the power of liquid chromatography-mass spectrometry-based urine metabolomics in negative ion mode by optimization of the additive.

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.

Bioactivity of sour cherry cultivars grown in Denmark.

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.

Bioactivity and chemical composition of blackcurrant (Ribes nigrum) cultivars with and without pesticide treatment.

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.

Relationship between the metabolite profile and technological properties of bovine milk from two dairy breeds elucidated by NMR-based metabolomics.

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.