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Only 6 to 8 % of the UK adults meet the daily recommendation for dietary fibre. Fava bean processing lead to vast amounts of high-fibre by-products such as hulls. Bean hull fortified bread was formulated to increase and diversify dietary fibre while reducing waste. This study assessed the bean hull: suitability as a source of dietary fibre; the systemic and microbial metabolism of its components and postprandial events following bean hull bread rolls. Nine healthy participants (53·9 ± 16·7 years) were recruited for a randomised controlled crossover study attending two 3 days intervention sessions, involving the consumption of two bread rolls per day (control or bean hull rolls). Blood and faecal samples were collected before and after each session and analysed for systemic and microbial metabolites of bread roll components using targeted LC-MS/MS and GC analysis. Satiety, gut hormones, glucose, insulin and gastric emptying biomarkers were also measured. Two bean hull rolls provided over 85 % of the daily recommendation for dietary fibre; but despite being a rich source of plant metabolites (P = 0·04 v. control bread), these had poor systemic bioavailability. Consumption of bean hull rolls for 3 days significantly increased plasma concentration of indole-3-propionic acid (P = 0·009) and decreased faecal concentration of putrescine (P = 0·035) and deoxycholic acid (P = 0·046). However, it had no effect on postprandial plasma gut hormones, bacterial composition and faecal short chain fatty acids amount. Therefore, bean hulls require further processing to improve their bioactives systemic availability and fibre fermentation.

Purpose This study evaluated the postprandial effects following consumption of buckwheat, fava bean, pea, hemp and lupin compared to meat (beef); focussing on biomarkers of satiety, gut hormones, aminoacids and plant metabolites bioavailability and metabolism. Methods Ten subjects (n = 3 men; n = 7 women; 42 ± 11.8 years of age; BMI 26 ± 5.8 kg/m2) participated in six 1-day independent acute interventions, each meal containing 30 g of protein from buckwheat, fava bean, pea, hemp, lupin and meat (beef). Blood samples were collected during 24-h and VAS questionnaires over 5-h. Results Volunteers consumed significantly higher amounts of most amino acids from the meat meal, and with few exceptions, postprandial composition of plasma amino acids was not significantly different after consuming the plant-based meals. Buckwheat meal was the most satious (300 min hunger scores, p p = 0.01) found after hemp compared with the other plant-based meals. Decreased plasma ghrelin concentrations (iAUC p p p Conclusion Plants are valuable sources of amino acids which are promoting satiety. The impact of hemp and buckwheat on GLP-1 and, respectively, BCAAs should be explored further as could be relevant for aid and prevention of chronic diseases such as type 2 diabetes. Study registered with clinicaltrial.gov on 12th July 2013, study ID number: NCT01898351.

New findings What is the central question of this study? Does exercise affect vitamin D receptor expression in T lymphocytes in young, middle-aged and older adults? What is the main finding and its importance? Moderate-intensity cycling exercise increases vitamin D receptor expression in vitamin D-deficient men, independent of age, presenting a strategy to combat the prevalence of vitamin D deficiency. Abstract Vitamin D plays a key role in the modulation of the immune system, mediated through the intracellular vitamin D receptor (VDR). Exercise has been shown to influence the activity and availability of the VDR. The aim of this study was to investigate the effect of age on basal immune cell (T-lymphocyte) VDR expression and the subsequent effect of acute aerobic exercise to modulate VDR expression in peripheral T cells. Thirty-five men were included in the study (mean ± SD: age 44 ± 17 years and body mass index 25.7 ± 3.1 kg/m2 ), separated into three age groups: 18-30 (n = 12), 31-45 (n = 11) and 60-75 years (n = 12). Participants completed two trials [control (CON) and aerobic exercise (AE)], with blood samples collected pre- and postexercise (0, 1 and 3 h). Peripheral blood T cells were isolated and analysed for VDR expression by flow cytometry. The results show that advanced age is associated with lower VDR expression in T cells (882 ± 274, 796 ± 243 and 594 ± 174 geomean in the 18-30, 31-45 and 60-75 year age groups, respectively). Acute AE was successful at acutely increasing VDR expression in T cells, irrespective of age. Advanced age corresponds to a lower T-cell VDR expression, which might be responsible for age-associated development of chronic conditions and autoimmunity. Exercise was successful in increasing VDR expression in T cells irrespective of age and independent of exercise-induced T-cell mobilization.

In this paper, we present an overview of how the International Space Station (ISS) safety engineering methodology directed to controlling extravehicular activity (EVA) crew electrical shock hazards, caused by ISS spacecraft charging, has evolved over the past 25+ years. Long-term measurements of ISS charging severity and frequency-of-occurrence, combined with detailed probabilistic analysis of EVA electric shock-circuit completion, led to a change in hazard control methodology. The requirement for two-fault tolerant EVA shock hazard control during all EVAs was replaced with a less operationally burdensome and risky EVA shock hazard detection and warning process. The applicability of event probability-based detection-and-warning processes to human spaceflight charging hazard control beyond low-earth orbit (LEO) is also considered.

Background Agitated or aggressive patients pose a high risk of emotional and physical harm to hospital staff. Healthcare associates have the highest rate of workplace violence among studied fields. Learning to effectively de-escalate a patient who is a danger to self or others is key to reducing these incidents. This course was designed to improve education and communication among staff on a hospital surgical floor when verbal de-escalation is needed. Methods A ninety-minute simulation-based multidisciplinary curriculum was developed. This included a 30-minute didactic discussion, 10-minute simulation, and 50-minute debrief. Learners included nurses, patient service associates, and protective services officers from a medical/surgical unit. Data were collected using a validated return on investment in learning protocol and the Management of Aggression and Violence Attitude Scale (MAVAS) tool. Results Our return on investment in learning showed that more than 97% of learners felt safer in managing agitated patients after participating in the training. The MAVAS tool was used in pre- and post-format and showed a significant trend toward the importance of clear communication and role clarity when de-escalating a patient. Conclusions A combination of didactic teaching and simulated experience allowed for greater confidence, communication, and teamwork in de-escalating an agitated patient in a community hospital surgical unit.

Cereal foods are commonly contaminated with multiple mycotoxins resulting in frequent human mycotoxin exposure. Children are at risk of high-level exposure because of their high cereal intake relative to body weight. Hence, this study aims to assess multimycotoxin exposure in UK children using urinary biomarkers. Spot urines (n = 21) were analyzed for multimycotoxins (deoxynivalenol, DON; nivalenol, NIV; ochratoxin A, OTA; zearalenone, ZEN; α-zearalenol, α-ZEL; β-zearalenol, β-ZEL; T-2 toxin, T-2; HT-2 toxin, HT-2; and aflatoxin B1 and M1, AFB1, AFM1) using liquid chromatography-coupled tandem mass spectrometry. Urine samples frequently contained DON (13.10 ± 12.69 ng/mL), NIV (0.36 ± 0.16 ng/mL), OTA (0.05 ± 0.02 ng/mL), and ZEN (0.09 ± 0.07 ng/mL). Some samples (1-3) contained T-2, HT-2, α-ZEL, and β-ZEL but not aflatoxins. Dietary mycotoxin estimation showed that children were frequently exposed to levels exceeding the tolerable daily intake (52 and 95% of cases for DON and OTA). This demonstrates that UK children are exposed to multiple mycotoxins through their habitual diet.

Fusarium mycotoxins are common contaminants in cereals and often co-occur with plant-derived mycotoxin sugar conjugates. Several of these modified mycotoxins are not degraded in the small intestine and hence carried through to the large intestine where microbial transformation may occur. This study aims to assess the gastrointestinal stability of the trichothecenes HT-2 toxin (HT-2), HT-2-β-glucoside (HT-2-Glc), diacetoxyscirpenol (DAS), DAS-α-glucoside (DAS-Glc) and fumonisin B1 (FB1), N-(1-deoxy-d-fructos-1-yl) fumonisin-B1 (NDF-FB1). All tested modified mycotoxins were stable under upper gastrointestinal (GI) conditions. In faecal batch culture experiments, HT-2-Glc was hydrolysed efficiently and no further microbial biotransformation of HT-2 was observed. DAS-Glc hydrolysis was slow and DAS was de-acetylated to 15-monoacetoxyscripenol. NDF-FB1 was hydrolysed at the slowest rate and FB1 accumulation varied between donor samples. Our results demonstrate that all tested modified mycotoxins are stable in the upper GI tract and efficiently hydrolysed by human gut microbiota, thus potentially contributing to colonic toxicity. Hence the microbial biotransformation of any novel modified mycotoxins needs to be carefully evaluated.

Mycotoxins are important food contaminants that commonly co-occur with modified mycotoxins such as mycotoxin-glucosides in contaminated cereal grains. These masked mycotoxins are less toxic, but their breakdown and release of unconjugated mycotoxins has been shown by mixed gut microbiota of humans and animals. The role of different bacteria in hydrolysing mycotoxin-glucosides is unknown, and this study therefore investigated fourteen strains of human gut bacteria for their ability to break down masked mycotoxins. Individual bacterial strains were incubated anaerobically with masked mycotoxins (deoxynivalenol-3-&beta, glucoside, DON-Glc, nivalenol-3-&beta, glucoside, NIV-Glc, HT-2-&beta, glucoside, HT-2-Glc, diacetoxyscirpenol-&alpha, glucoside, DAS-Glc), or unconjugated mycotoxins (DON, NIV, HT-2, T-2, and DAS) for up to 48 h. Bacterial growth, hydrolysis of mycotoxin-glucosides and further metabolism of mycotoxins were assessed. We found no impact of any mycotoxin on bacterial growth. We have demonstrated that Butyrivibrio fibrisolvens, Roseburia intestinalis and Eubacterium rectale hydrolyse DON-Glc, HT-2 Glc, and NIV-Glc efficiently and have confirmed this activity in Bifidobacterium adolescentis and Lactiplantibacillus plantarum (DON-Glc only). Prevotella copri and B. fibrisolvens efficiently de-acetylated T-2 and DAS, but none of the bacteria were capable of de-epoxydation or hydrolysis of &alpha, glucosides. In summary we have identified key bacteria involved in hydrolysing mycotoxin-glucosides and de-acetylating type A trichothecenes in the human gut.

Benefits from micronutrients within fruit juice and smoothies are well documented, but fewer studies research the role of phytochemicals. Well-controlled human studies are essential to evaluate their impact, particularly on glucose and lipid regulation but also gastrointestinal health. Planning these studies requires data on the potential molecular targets. Here we report a comprehensive metabolomic (LC-MS) analysis of the phytochemical composition of four commonly consumed beverages, including data on whether they are free to be absorbed early in the gastrointestinal tract or bound to other plant components. Smoothies contained a wide range of phenolics (free and bound), whereas the fruit juices contained higher amounts of fewer compounds. Orange juice was rich in bound hesperidin (1.97 &plusmn, 0.39 mg/100 mL) and hydroxycinnamic acids, likely to be delivered to the colon with the potential to have an impact on gut health. Apple juice contained free chlorogenic acid (3.11 &plusmn, 1.03 mg/100 mL), phloridzin (0.40 &plusmn, 0.03 mg/100 mL), catechin (0.090 &plusmn, 0.005 mg/100 mL), and epicatechin (0.38 &plusmn, 0.02 mg/100 mL), suggesting potential roles in glucose uptake reduction or positive effects on systemic blood flow. Redox screening established that differences in chemical composition impacted on bioactivity, highlighting the importance of availability from the matrix. This suggests that fruit-based beverage interventions should target specific mechanisms depending on the fruits from which they are comprised and in particular, the availability of the individual constituents.

Legumes are a source of health-promoting macro- and micronutrients, but also contain numerous phytochemicals with useful biological activities, an example of which are saponins. Epidemiological studies suggest that saponins may play a role in protection from cancer and benefit human health by lowering cholesterol. Therefore, they could represent good candidates for specialised functional foods. Following the consumption of a soya-rich high-protein weight-loss diet (SOYA HP WL), the concentrations of Soyasaponin I (SSI) and soyasapogenol B (SSB) were determined in faecal samples from human volunteers (n = 10) and found to be between 1.4 and 17.5 mg per 100 g fresh faecal sample. SSB was the major metabolite identified in volunteers’ plasma (n = 10) after consumption of the soya test meal (SOYA MEAL); the postprandial (3 h after meal) plasma concentration for SSB varied between 48.5 ng/mL to 103.2 ng/mL. The metabolism of SSI by the gut microbiota (in vitro) was also confirmed. This study shows that the main systemic metabolites of soyasaponin are absorbed from the gut and that they are bioavailable in plasma predominantly as conjugates of sapogenol. The metabolism and bioavailability of biologically active molecules represent key information necessary for the efficient development of functional foods.

Apios americana Medik, a native American plant has potential as a commercially viable Northern European-grown rootcrop, mainly due to its resistance to extreme climate and nutritional quality. Analysis of A. americana sourced from two UK sites; South (51.4690 °N, 1.1150 °W) and North (55.9661 °N, 3.2063 °W) showed that the tubers were a complete source of amino acids (UPLC-TUV analysis), were rich in protein (15.0 ± 0.0160 and 17.3 ± 0.0779%; Vario Max CN analysis), fibre (total non-starch polysaccharides, 10.4 ± 0.570 and 10.6 ± 0.280%; GC analysis) and micronutrients (calcium, manganese, iron, zinc, molybdenum, potassium and phosphorus; ICP-MS analysis). Apios americana tubers were also rich in bioactive phytochemicals. From the 156 plant metabolites measured using LC-MS/MS analysis, genistein was the major phytophenol in both the Southern- and Northern UK tubers (259 ± 12.2 mg Kg−1 and 356 ± 29.9 mg Kg−1 respectively); the peel having similar phytochemical profiles. The protein and fibre content of the leaves (17.3 ± 0.0434% and 11.7 0.0445%) and rhizomes (18.4 ± 0.0152% and 13.5 ± 0.590%) were significantly higher (p

The hypoglycaemic effects of two Ribes sp. i.e., anthocyanin-rich black currants (BC) were compared to green currants (GC), which are low in anthocyanins to establish which compounds are involved in the regulation of postprandial glycaemia. We determined the effect of the currants on inhibiting carbohydrate digestive enzymes (α-amylase, α-glucosidase), intestinal sugar absorption and transport across CaCo-2 cells. The digestion of these currants was modelled using in vitro gastrointestinal digestion (IVGD) to identify the metabolites present in the digested extracts by LC-MS/MS. Freeze-dried BC and IVDG extracts inhibited yeast α-glucosidase activity (P

Using simple solvent extraction and enzymatic hydrolysis, a rapid LC-MS/MS method for quantification of free and conjugated forms of anthocyanidins and anthocyanins in plasma and urine samples was developed and validated. A mixed enzymatic treatment containing β-glucuronidase (100 U mL

Mycotoxin contamination of cereal grains causes well-recognized toxicities in animals and humans, but the fate of plant-bound masked mycotoxins in the gut is less well understood. Masked mycotoxins have been found to be stable under conditions prevailing in the small intestine but are rapidly hydrolyzed by fecal microbiota. This study aims to assess the hydrolysis of the masked mycotoxin deoxynivalenol-3-glucoside (DON3Glc) by the microbiota of different regions of the porcine intestinal tract. Intestinal digesta samples were collected from the jejunum, ileum, cecum, colon, and feces of 5 pigs and immediately frozen under anaerobic conditions. Sample slurries were prepared in M2 culture medium, spiked with DON3Glc or free deoxynivalenol (DON; 2 nmol/ml), and incubated anaerobically for up to 72 h. Mycotoxin concentrations were determined using liquid chromatography-tandem mass spectrometry, and the microbiota composition was determined using a quantitative PCR methodology. The jejunal microbiota hydrolyzed DON3Glc very slowly, while samples from the ileum, cecum, colon, and feces rapidly and efficiently hydrolyzed DON3Glc. No further metabolism of DON was observed in any sample. The microbial load and microbiota composition in the ileum were significantly different from those in the distal intestinal regions, whereas those in the cecum, colon and feces did not differ. IMPORTANCE Results from this study clearly demonstrate that the masked mycotoxin DON3Glc is hydrolyzed efficiently in the distal small intestine and large intestine of pigs. Once DON is released, toxicity and absorption in the distal intestinal tract likely occur in vivo . This study further supports the need to include masked metabolites in mycotoxin risk assessments and regulatory actions for feed and food.

Reformulation of existing processed food or formulation of new foods using natural products (plant-based) will inherently confer to new products with less calories, fat, salt, phosphates and other synthetic components, and higher amounts of fibre, antioxidants, vitamins and other beneficial components. Plant ingredients, such as food plant powders, are currently being used in food manufacturing, predominantly for flavouring and colouring purposes. To expand their use as a food ingredient, freeze-dried powders representing major vegetable groups were characterised by targeted LC-MS/MS analysis of their phytochemicals. All the plant powders were found to be rich in flavonoids, phenolic acids and derivatives; total content in these compounds varied from around 130 mg kg(-1) (green pea) to around 930 mg kg(-1) (spinach). The food plant powders' phytochemical content represents valuable information for the food industry in the development of healthier novel foods and for the reformulation of existing food products in relation to antioxidants, food preservatives and alternatives to nitrite use.

Patterns of DNA methylation (5-methylcytosine, 5mC) are rearranged during differentiation contributing to the regulation of cell type-specific gene expression. TET proteins oxidise 5mC to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC). Both 5fC and 5caC can be recognised and excised from DNA by thymine-DNA glycosylase (TDG) followed by the subsequent incorporation of unmodified cytosine into the abasic site via the base excision repair (BER) pathway. We previously demonstrated that 5caC accumulates during lineage specification of neural stem cells (NSCs) suggesting that such active demethylation pathway is operative in this system, however it is still unknown if TDG/BER-dependent demethylation is utilised during other types of cellular differentiation. Here we analyse dynamics of the global levels of 5hmC and 5caC during differentiation of human pluripotent stem cells (hPSCs) towards hepatic endoderm. We show that, similar to differentiating NSCs, 5caC transiently accumulates during hepatic differentiation. The levels of 5caC increase during specification of foregut, peak at the stage of hepatic endoderm commitment and drop in differentiating cells concurrently with the onset of expression of Alpha Fetoprotein, a marker of committed hepatic progenitors. Moreover, we show that 5caC accumulates at promoter regions of several genes expressed during hepatic specification at differentiation stages corresponding to the commencement of their expression. Our data indicate that transient 5caC accumulation is a common feature of two different types (neural/glial and endoderm/hepatic) of cellular differentiation. This suggests that oxidation of 5mC may represent a general mechanism of rearrangement of 5mC profiles during lineage specification of somatic cells in mammals.

Dietary exposure to deoxynivalenol (DON) has been reported previously in the UK, but levels were low and most individuals are well protected by the maximum permitted levels in food set by the European Commission. However, no information is available on annual fluctuation in dietary DON exposure. We hypothesised that dietary DON exposure may vary when individuals consume cereals derived from harvests with low (2011) and high (2012) Fusarium prevalence. In this pilot study, spot urine samples were collected in years 1 and 2 from 15 volunteers following their habitual diet. Urinary DON was analysed by LC-MS/MS to estimate 24-h DON excretion and daily dietary DON intake. DON was detectable in all urine samples with an average excretion of 10.08 ± 9.13 µg/24-h urine in year 1 which significantly (p = 0.005) increased to 24.84 ± 13.83 µg/24-h urine in year 2. This resulted from an estimated dietary intake of 195.94 ± 166.44 ng DON kg(-1) BW in year 1 and 518.64 ± 292.49 ng DON kg(-1) BW in year 2. Based on these estimates, the tolerable daily intake for DON was exceeded in 13% of occasions in year 2 and none in year 1. This pilot study is based on estimates of DON intake derived from urinary DON excretion. Results suggest that DON exposure varies annually and that current maximum levels might not sufficiently protect consumers during years of high Fusarium prevalence.

Scope Plant secondary metabolites, such as phenolic acids are commonly associated with benefits for human health. Two of the most abundant phenylpropanoid-derived compounds detected in human faecal samples are phenylacetic acid (PAA) and 4-hydroxylphenylacetic acid (4-hydroxyPAA). Although they have the potential to be derived from diets rich in plant-based foods, evidence suggests that these compounds can be derived from the microbial fermentation of aromatic amino acids (AAAs) in the colon. Methods and results To identify the bacteria responsible, 26 strains representing 25 of the dominant human colonic species were screened for phenyl metabolite formation. Seven strains produced significant amounts of both PAA and 4-hydroxyPAA. These included five out of seven Bacteroidetes (Bacteroides thetaiotaomicron, Bacteroides eggerthii, Bacteroides ovatus, Bacteroides fragilis, Parabacteroides distasonis), and two out of 17 Firmicutes (Eubacterium hallii and Clostridium bartlettii). These species also produced indole-3-acetic acid (IAA), the corresponding tryptophan metabolite, but C. bartlettii showed 100 times higher IAA production than the other six strains. Four strains were further tested and PAA formation was substantially increased by phenylalanine, 4-hydroxyPAA by tyrosine and IAA by tryptophan. Conclusion This study demonstrates that certain microbial species have the ability to ferment all three AAAs and that protein fermentation is the likely source of major phenylpropanoid-derived metabolites in the colon.

Sustainable sources of high-protein plants could help meet future protein requirements. Buckwheat, green pea, fava bean, hemp, and lupin were analyzed by proximate analysis and inductively coupled plasma mass spectrometry to determine their macro- and micronutrient contents, and liquid chromatography–mass spectrometry was used to elucidate the phytochemical profiles. The protein contents ranged from 20 to 43% (w/w), and all samples were found to be rich in insoluble fiber: 9–25% (w/w). The selected crops had a favorable micronutrient profile, with phosphorus levels ranging from 2.22 ± 0.05 to 9.72 ± 0.41 g kg–1, while iron levels ranged from 20.23 ± 0.86 to 69.57 ± 7.43 mg kg–1. The crops contained substantial amounts of phytophenolic compounds. In particular, buckwheat was a rich source of pelargonidin (748.17 ± 75.55 mg kg–1), epicatechin (184.1 ± 33.2 mg kg–1), quercetin (35.66 ± 2.22 mg kg–1), caffeic acid (41.74 ± 22.54 mg kg–1), and 3-hydroxyphenylacetic acid (63.64 ± 36.16 mg kg–1); hemp contained ...

A methyl-deficient diet (MD) lacking folic acid and the associated methyl donors choline and methionine, fed to the laboratory rat during the periods of oocyte and embryo development, has been shown to programme glucose metabolism in the offspring. The hepatic proteome of the male offspring of female rats fed MD diets for 3 weeks prior to mating and for the first 5 days of gestation has been examined by 2-dimensional gel electrophoresis. Three groups of differentially abundant proteins associated with energy metabolism, amino acid metabolism and antioxidant defence were identified in the soluble proteins extracted from the liver from the MD offspring at both 6 and 12 months of age. Altered mitochondrial activity in other programming models leads to a similar pattern of differential protein abundance. Two of the differentially abundant proteins were identified as GAPDH and PGK-1 by mass spectrometry. Western blotting showed that there were multiple isoforms of both proteins with similar molecular weights but different isoelectric points. The differentially abundant spots reduced in the MD offspring corresponded to minor isoforms of GAPDH and PGK-1. The levels of PPAR-alpha, SREBP and glucocorticoid receptor mRNAs associated with other models of prenatal programming were unchanged in the MD offspring. The data suggest that a diet deficient in folic acid and associated methyl donors fed during the peri-conception and early preimplantation periods of mammalian development affects mitochondrial function in the offspring and that the posttranslational modification of proteins may be important.

Purpose: Platelets play a key role in haemostasis and wound healing, contributing to formation of vascular plugs. They are also involved in formation of atherosclerosic plaques. Some traditional diets, like the Mediterranean diet, are associated with a lower risk of cardiovascular disease. Components in these diets may have anti-platelet functions contributing to their health benefits. Methods: We studied the effects of alperujo extract, an olive oil production waste product containing the majority of polyphenols found in olive fruits, through measurement of effects on platelet aggregation and activation in isolated human platelets, and through identification of changes in the platelet proteome. Results: Alperujo extract (40 mg/L) significantly decreased in vitro ADP- (p = 0.002) and TRAP- (p = 0.02) induced platelet activation as measured by the flow cytometry using the antibody for p-selectin (CD62p), but it did not affect the conformation of the fibrinogen receptor as measured by flow cytometry using the antibodies for anti-fibrinogen, CD42a and CD42b. Alperujo extract (100 mg/L) inhibited both collagen- and TRAP-induced platelet aggregation by 5% (p < 0.05), and a combination of hydroxytyrosol and 3,4-dihydroxyphenylglycol were, at least partly, responsible for this effect. Proteomic analysis identified nine proteins that were differentially regulated by the alperujo extract upon ADP-induced platelet aggregation. These proteins represent important mechanisms that may underlie the anti-platelet effects of this extract: regulation of platelet structure and aggregation, coagulation and apoptosis, and signalling by integrin αIIb/β3. Conclusions: Alperujo extract may protect against platelet activation, platelet adhesion and possibly have anti-inflammatory properties. © 2010 Springer-Verlag.

Despite fruit, vegetables and many processed products counting towards achieving the recommended five-a-day strategy, it is inevitable that produce choice will affect the benefits delivered. Fruits locally produced and commonly consumed in Scotland were compared for their phenolic acid content and form. The phenolic acid composition was highly variable, but the locally produced fruits were significantly ( p – 4.89 g/kg compared to 0.06 – 0.22 g/kg). The majority of the phenolic acids were conjugated to other plant components, suggesting that any health benefits derived from these compounds are likely to be after they are released/metabolised by the colonic microbiota. Although the potential protective effects of the individual compounds will not be ascertained until the exact role of these compounds in disease prevention has been clarified, it is clear that the total amount of phenolic acids in the diet will vary enormously depending on the types of fruits consumed.

The development of insulin resistance in the obese is associated with chronic, low-grade inflammation. We aimed to identify novel links between obesity, insulin resistance and the inflammatory response by comparing C57BL/6 with type I interleukin-1 receptor knockout (IL-1RI(-/-)) mice, which are protected against diet-induced insulin resistance. Mice were fed a high-fat diet for 16 wk. Insulin sensitivity was measured and proteomic analysis was performed on adipose, hepatic and skeletal muscle tissues. Despite an equal weight gain, IL-1RI(-/-) mice had lower plasma glucose, insulin and triacylglycerol concentrations, compared with controls, following dietary treatment. The higher insulin sensitivity in IL-1RI(-/-) mice was associated with down-regulation of antioxidant proteins and proteasomes in adipose tissue and hepatic soluble epoxide hydrolase, consistent with a compromised inflammatory response as well as increased glycolysis and decreased fatty acid beta-oxidation in their muscle. Their lower hepatic triacylglycerol concentrations may reflect decreased flux of free fatty acids to the liver, decreased hepatic fatty acid-binding protein expression and decreased lipogenesis. Correlation analysis revealed down-regulation of classical biomarkers of ER stress in their adipose tissue, suggesting that disruption of the IL-1RI-mediated inflammatory response may attenuate cellular stress, which was associated with significant protection from diet-induced insulin resistance, independent of obesity.

Foetal growth is particularly sensitive to the protein content of the mother's diet. Microarray data from the foetal liver of pregnant rats fed normal (HP) or reduced protein diets (LP) were compared by gene set enrichment analysis. Soluble proteins from a second portion of the liver were analysed by two-dimensional gel electrophoresis. Genes associated with progesterone, insulin-like growth factor-1 and vascular endothelial growth factor were upregulated in HP compared to LP, in addition to genes associated with cell differentiation and signalling from the extracellular matrix. In contrast, cytokine signalling was downregulated. Proteomics showed that proteins associated with amino acid metabolism, mitochondrial function and cell motility were differentially abundant in the HP compared to the LP groups. These growth factor and extracellular matrix signalling pathways linked to cell motility may be important mediators of the changes in liver structure that occur in utero and persist into adult life.

Low folate intake is associated with colon cancer. We combined a proteomics and biochemical approach to identify proteins and pathways affected by folate deficiency in human colonocytes. Folate differentially altered activity and expression of proteins involved in proliferation [e.g., PCNA], DNA repair [e.g., XRCC5, MSH2], apoptosis [e.g., BAG family chaperone protein, DIABLO and porin], cytoskeletal organization [e.g., actin, ezrin, elfin], and expression of proteins implicated in malignant transformation [COMT, Nit2].

The importance of folic acid and the methionine cycle in fetal development is well recognised even though the mechanism has not been established. Since the cycle is active in the maternal liver, poor folate status may modify hepatic metabolism. Pregnant rats were fed diets deficient in folic acid (–F) or in three key methyl donors, folic acid, choline and methionine (–FLMLC) and the maternal liver was analysed on day 21 of gestation. Two-dimensional gel electrophoresis of soluble proteins identified differentially abundant proteins, which could be allocated into nine functional groups. Five involved in metabolic processes, namely, folate/methionine cycle, tyrosine metabolism, protein metabolism, energy metabolism and lipid metabolism, and three in cellular processes, namely, endoplasmic reticulum function, bile production and antioxidant defence. The mRNA for sterol regulatory element-binding protein-1c and acetyl-CoA carboxylase-1 (fatty acid synthesis) were decreased by both –F and –FLMLC diets. The mRNA for PPARα and PPARγ and carnitine palmitoyl transferase (fatty acid oxidation) were increased in the animals fed the –FLMLC diets. Changes in the abundance of proteins associated with intracellular lipid transport suggest that folate deficiency interferes with lipid export. Reduced fatty acid synthesis appeared to prevent steatosis in animals fed the –F diet. Even with increased oxidation, TAG concentrations were approximately three-fold higher in animals fed the –FLMLC diet and were associated with an increase in the relative abundance of proteins associated with oxidative stress. Fetal development may be indirectly affected by these changes in hepatic lipid metabolism.

We assessed the effects of Picual and Arbequina olive oil, rich and poor in polyphenols, respectively, on plasma lipid and glucose metabolism, hepatic fat content, and the hepatic proteome in female Apoe-/- mice. Both olive oils increased hepatic fat content and adipophilin levels (p < 0.05), though Picual olive oil significantly decreased plasma triglycerides (p < 0.05). Proteomics identified a range of hepatic antioxidant enzymes that were differentially regulated by both olive oils as compared with palm oil. We found a clear association between olive oil consumption and differential regulation of adipophilin and betaine homocysteine methyl transferase as modulators of hepatic triglyceride metabolism. Therefore, our "systems biology" approach revealed hitherto unrecognized insights into the triglyceride-lowering and anti-atherogenic mechanisms of extra virgin olive oils, wherein the up-regulation of a large array of anti-oxidant enzymes may offer sufficient protection against lesion development and diminish oxidative stress levels instigated by hepatic steatosis.

Cycle-by-cycle data have been shown to be effective in the analysis of a signalized intersection with measures of effectiveness such as volume-to-capacity ratios, arrival type, and average vehicular delay. Currently, actuated traffic controllers are unable to store vehicle counts and vehicle occupancy in cycle-by-cycle bins, requiring extra equipment and personnel to collect data in these bins. The objective of this research was to develop an integrated general purpose data collection module that time-stamps detector and phase state changes within a National Electrical Manufacturers Association actuated traffic signal controller and uses those data to provide quantitative graphs to assess arterial progression, phase capacity utilization, movement delay, and served volumes on a cycle-by-cycle basis. Given that the United States recently received a grade of 61 of 100 on the National Traffic Signal Report Card, it is particularly important that procedures such as these be used to provide performance measures over extended periods so agencies have cost-effective mechanisms for assessing and priority-ranking signal timing efforts.

The dietary phenolic compound, salicylic acid, decreases oxidative stress and pro-inflammatory and potentially neo-plastic prostaglandins with a concomitant increase in glutathione peroxidase activity. Salicylic acid, a dietary plant-based phenolic compound and also the main metabolite of aspirin, may contribute to the colon protective effects of plant-based diets. Oxidative stress is a characteristic of pre-cancerous and cancerous colon and inflammatory bowel diseases (IBD) that increase colon cancer risk. The mechanism(s) whereby salicylic acid modulates potentially pro-cancerous activity associated with oxidative stress is further investigated here using a proteomic approach. A rat model of oxidative stress was supplemented with salicylic acid (1 mg/kg diet, mean plasma levels 310+/-32 micromol/l). Soluble colon protein extracts were subjected to 2D PAGE. Salicylic acid modulated proteins, identified using MALDI-TOF and LC/MS/MS, are involved in protein folding, transport, redox, energy metabolism and cytoskeletal regulation. A partial least squares (PLS) analysis approach was used to assist biological interpretation of the altered protein profiles via their associations between previously published biochemical measurements of oxidative stress, prostaglandin levels and increased glutathione peroxidase activity. Early detection of altered homeostasis in colon may assist in identifying pre-pathological features preceding colon tumorigenesis and contribute to an understanding of epidemiological evidence supporting a protective effect of plant-based diets.

Bacterial butyryl-CoA CoA-transferase activity plays a key role in butyrate formation in the human colon, but the enzyme and corresponding gene responsible for this activity have not previously been identified. A novel CoA-transferase gene is described from the colonic bacteriumRoseburiasp. A2-183, with similarity to acetyl-CoA hydrolase as well as 4-hydroxybutyrate CoA-transferase sequences. The gene product, overexpressed in anEscherichia colilysate, showed activity with butyryl-CoA and to a lesser degree propionyl-CoA in the presence of acetate. Butyrate, propionate, isobutyrate and valerate competed with acetate as the co-substrate. Despite the sequence similarity to 4-hydroxybutyrate CoA-transferases, 4-hydroxybutyrate did not compete with acetate as the co-substrate. Thus the CoA-transferase preferentially uses butyryl-CoA as substrate. Similar genes were identified in other butyrate-producing human gut bacteria from clostridial clusters IV and XIVa, while other candidate CoA-transferases for butyrate formation could not be detected inRoseburiasp. A2-183. This suggests strongly that the newly identified group of CoA-transferases described here plays a key role in butyrate formation in the human colon.

Background: The objective of the present study was to examine the outcome of nasal fracture reduction under both local anaesthesia and general anaesthesia techniques. Methods: Patient records were obtained from clinic lists of patients attending the otolaryngology (ORL) outpatient unit at ­Wellington Hospital or the plastic surgical unit at Hutt Hospital over a 24-month period. To meet criteria for inclusion, patients had to have an isolated nasal deformity, be aged 14 years or older and be at least 6 months post reduction. Functional and aesthetic results were assessed using subjective and objective end outcome measures. A questionnaire was developed and used to assess subjective outcomes. The number of patients proceeding to open operative correction was used for the objective measure, which included septoplasty, septorhinoplasty or rhinoplasty. Results: A total of 197 patients had nasal fractures manipulated over this period. One hundred and seven were manipulated under general anaesthesia (GA) and 90 were manipulated under local anaesthesia (LA). The questionnaire completion rate was 66%. Patient demographics were statistically similar between the two groups. There was no statistical difference in patients" subjective assessment of outcome with treatment by GA or LA. More patients preferred to have their fracture manipulated under GA but this was not ­statistically significant. There was no significant difference between GA and LA groups in progression to open operative correction. Conclusions: Both LA and GA appear to be acceptable and satisfactory methods of anaesthesia in the reduction of simple displaced nasal fracture.

Cereal grains are commonly contaminated with Fusarium mycotoxins and their plant-derived masked metabolites. The fate of masked mycotoxins in the human gut is poorly understood. Here we assess the metabolism and transport of glucoside metabolites of common trichothecenes (deoxynivalenol, nivalenol, T-2 toxin) and zearalenone compounds (zearalenone, α- and β-zearalenol) in the human gut in vitro.Masked mycotoxins were incubated with artificial digestive juices and absorption was assessed in differentiated Caco-2/TC7 cells. Colonic metabolism was studied using fecal batch cultures from five donors and mycotoxins were detected using LC-MS/MS. All masked mycotoxins were stable under upper GI tract conditions and no absorption was observed. Free trichothecenes were absorbed intact whereas free zearalenone compounds were absorbed and metabolized to undetected compounds by Caco-2/TC7 cells. Human gut microbiota efficiently hydrolyzed all masked mycotoxins. Trichothecenes were fully recovered as parent mycotoxins whereas 40-70% of zearalenone compounds were further metabolized to unknown metabolites.Our results demonstrate that masked trichothecenes will reach the colon intact to be released as parent mycotoxins by gut microbiota, hence contributing to mycotoxin exposure. Masked zearalenone compounds are metabolized by gut microbiota and epithelial cells and the identity and toxicity of metabolites remain to be determined.

Phytophenols present in cereals are metabolised to compounds that could be partly responsible for the reduced risk of chronic diseases and all-cause mortality associated with fibre-rich diets. The bioavailability, form and in vivo concentrations of these metabolites require to be established.Eight healthy volunteers consumed a test meal containing a recommended dose (40 g) and high dose (120 g) of ready-to-eat wheat bran cereal and the systemic and colonic metabolites determined quantitatively by LC-MS.Analysis of the systemic metabolomes demonstrated that a wide range of phytophenols were absorbed/excreted (43 metabolites) within 5 h of consumption. These included 16 of the 21 major parent compounds identified in the intervention product and several of these were also found to be significantly increased in the colon. Not all of the metabolites were increased with the higher dose, suggesting some limitation in absorption due to intrinsic factors and/or the food matrix. Many compounds identified (e.g. ferulic acid and major metabolites) exhibit anti-inflammatory activity and impact on redox pathways. The combination of postprandial absorption and delivery to the colon, as well as hepatic recycling of the metabolites at these concentrations, is likely to be beneficial to both systemic and gut health.

Low B vitamin status is linked with human vascular disease. We employed a proteomic and biochemical approach to determine whether nutritional folate deficiency and/or hyperhomocysteinemia altered metabolic processes linked with atherosclerosis in ApoE null mice. Animals were fed either a control fat (C; 4 % w/w lard) or a high-fat [HF; 21 % w/w lard and cholesterol (0/15 % w/w)] diet with different B vitamin compositions for 16 weeks. Aorta tissue was prepared and global protein expression, B vitamin, homocysteine and lipoprotein status measured. Changes in the expression of aorta proteins were detected in response to multiple B vitamin deficiency combined with a high-fat diet (P

Saprolegnia parasitica is a freshwater oomycete that is capable of infecting several species of fin fish. Saprolegniosis, the disease caused by this microbe, has a substantial impact on Atlantic salmon aquaculture. No sustainable treatment against saprolegniosis is available, and little is known regarding the host response. In this study, we examined the immune response of Atlantic salmon to S. parasitica infection and to its cell wall carbohydrates. Saprolegnia triggers a strong inflammatory response in its host (i.e., induction of interleukin-1β 1 [IL-1β 1 ], IL-6, and tumor necrosis factor alpha), while severely suppressing the expression of genes associated with adaptive immunity in fish, through downregulation of T-helper cell cytokines, antigen presentation machinery, and immunoglobulins. Oomycete cell wall carbohydrates were recognized by fish leukocytes, triggering upregulation of genes involved in the inflammatory response, similar to what is observed during infection. Our data suggest that S. parasitica is capable of producing prostaglanding E 2 (PGE 2 ) in vitro , a metabolite not previously shown to be produced by oomycetes, and two proteins with homology to vertebrate enzymes known to play a role in prostaglandin biosynthesis have been identified in the oomycete genome. Exogenous PGE 2 was shown to increase the inflammatory response in fish leukocytes incubated with cell wall carbohydrates while suppressing genes involved in cellular immunity (gamma interferon [IFN-γ] and the IFN-γ-inducible protein [γ-IP]). Inhibition of S. parasitica zoospore germination and mycelial growth by two cyclooxygenase inhibitors (aspirin and indomethacin) also suggests that prostaglandins may be involved in oomycete development.

Mammalian metallothioneins (MT), are characteristically N(alpha)-acetylated and the presence of an unblocked N-terminus has not previously been reported. On-line capillary electrophoresis-electrospray mass spectrometry of hepatic MT-2 from rats injected with zinc revealed two isoforms differing by a mass equivalent to that of a single acetyl group. The lower mass component constituted > 20% of total MT-2 protein and both MT-2 isoforms were separated by reversed-phase high-performance liquid chromatography. The identity of each fraction was confirmed by matrix-assisted laser desorption ionisation mass spectrometry, and amino acid analysis and N-terminal sequencing revealed that the lower mass isoform was unblocked at the N-terminus and had an amino acid composition and sequence which is characteristic of rat MT-2. Thus the complementary techniques of mass spectrometry and N-terminal sequencing demonstrated conclusively that purified MT-2 from zinc-treated rats contains an unacetylated isoform. We propose that the cotranslational acetylation of rat MT-2 may under some circumstances be inefficient compared to that in other nonrodent species, where we have detected only trace levels of unacetylated MT isoforms.

Conjugated linoleic acids (CLA) affect atherogenesis, but mechanisms are not well understood. We explored how two isomers of CLA, cis9, trans11-CLA and trans10, cis12-CLA, affected lipid and glucose metabolism, as well as hepatic protein expression, in apolipoprotein E knockout mice. After 12 wk of intervention, plasma triglyceride, NEFA, and glucose concentrations were significantly higher in the trans10, cis12-CLA group, whereas plasma triglyceride, NEFA, glucose, and insulin concentrations were significantly lower in the cis9, trans11-CLA group, compared with control mice consuming linoleic acid. Proteomics identified significant up- or down-regulation of 113 liver cytosolic proteins by either CLA isomer. Principal component analysis revealed that the treatment effect of cis9, trans11-CLA was mainly explained by the up-regulation of different posttranslational forms of heat shock protein 70 kD. In contrast, the treatment effect of trans10, cis12-CLA was mainly explained by up-regulation of key enzymes in the gluconeogenic, beta-oxidation, and ketogenesic pathways. Correlation analysis again emphasized the divergent effects of both CLA isomers on different pathways, but also revealed a linkage between insulin resistance and increased levels of hepatic serotransferrin. Thus, our systems biology approach provided novel insights into the mechanisms by which individual CLA isomers differentially affect pathways related to atherogenesis, such as insulin resistance and inflammation.

Whole-grain diets are linked to reduced risk of several chronic diseases (heart disease, cancer, diabetes, metabolic syndrome) and all-cause mortality. There is increasing evidence that these benefits are associated with the gut microbiota and that release of fibre-related phenolic metabolites in the gut is a contributing factor. Additional sources of these metabolites include fruits and vegetables, but the evidence for their protective effects is less well established. With respect to the availability of bound phytophenols, ready-to-eat cereals are compared with soft fruits (considered rich in antioxidants) and other commonly consumed fruits and vegetables. The results demonstrated that when compared with an equivalent serving of fruits or vegetables, a recommended portion of whole-grain cereals deliver substantially higher amounts of bound phytophenols, which are available for metabolism in the colon. The increased amount of these phenolic metabolites may, in part, explain the evidence for the protective effects of whole-grain cereals.