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Bioactive compounds including anthocyanins and other polyphenols are associated with reduced lung inflammation and improved lung function in asthma and other lung diseases. This study investigated the effects of a Boysenberry and apple juice concentrate, high in cyanidin glycosides, ellagitannins, and chlorogenic acid, on a mouse model of allergic airways inflammation. Male C57BL/6J mice were orally gavaged with 2.5 mg/kg of total anthocyanins (TAC) from BerriQi® Boysenberry and apple juice concentrate (0.2 mg/kg human equivalent dose) or water control 1 hr before an acute intranasal ovalbumin (OVA) challenge and were gavaged again 2 days after the intranasal challenge. Consumption of BerriQi® Boysenberry and apple juice concentrate significantly decreased OVA‐induced infiltrating eosinophils, neutrophils, and T cells in the lung, and mucous production. Quantification of gene expression for arginase (Arg1), chitinase 3‐like 3 (Ym‐1), found in inflammatory zone (Fizz1), which have been associated with an anti‐inflammatory macrophage phenotype (M2), found significantly increased Arg1 expression in the lung in the Boysenberry and apple juice concentrate treatment group. There was also increased production of M2‐associated cytokines C‐X‐C motif chemokine ligand (CXCL) 10 and C‐C motif chemokine ligand (CCL) 4. These results suggest that consumption of BerriQi® Boysenberry and apple juice concentrate promoted a shift toward an anti‐inflammatory environment within the lung leading to reduced immune cell infiltration and tissue damage., We identified the polyphenols in a Boysenberry and apple juice concentrate and then investigated whether consumption inhibited immune cell infiltration into the lung in response to an allergen challenge in mice. We found Boysenberry and apple juice concentrate decreased immune cell infiltration and increased anti‐inflammatory genes and proteins suggesting that Boysenberry and apple juice concentrate mediated a switch to a more anti‐inflammatory lung environment leading to reduced tissue damage.

The groups of plant flavonoid metabolites termed anthocyanins and proanthocyanins (PA) are responsible for pigmentation in seeds, flowers and fruits. Anthocyanins and PAs are produced by a pathway of enzymes which are transcriptionally regulated by transcription factors (TFs) that form the MYB-bHLH-WD40 (MBW) complex. In this study, transcriptomic analysis of purple-pigmented kiwifruit skin and flesh tissues identified MYBC1, from subgroup 5 of the R2R3 MYB family, and WRKY44 (highly similar to Arabidopsis TTG2) as candidate activators of the anthocyanin pathway. Transient over-expression of MYBC1 and WRKY44 induced anthocyanin accumulation in tobacco leaves. Dual luciferase promoter activation assays revealed that both MYBC1 and WRKY44 were able to strongly activate the promoters of the kiwifruit F3′H and F3′5′H genes. These enzymes are branch points of the pathway which specifies the type of anthocyanin accumulated. Stable over-expression of MYBC1 and WRKY44 in kiwifruit calli activated the expression of F3′5′H and PA-related biosynthetic genes as well as increasing levels of PAs. These results suggest that while previously characterised anthocyanin activator MYBs regulate the overall anthocyanin biosynthesis pathway, the PA-related TFs, MYBC1 and WRKY44, more specifically regulate key branch points. This adds a layer of regulatory control that potentially balances anthocyanin and PA levels.

Background and aims: This cross-sectional observational case-control study was initiated in July 2016 with the aim of increasing an understanding of the underlying disease mechanisms in functional gastrointestinal disorders (FGIDs) including irritable bowel syndrome (IBS), functional diarrhoea (FD), and functional constipation (FC). Specific areas of interest include the effect of food, microbiome, host and microbial genetics, metabolome, and psychological variables on unexplained chronic gastrointestinal (GI) symptoms. Methods: This study recruited consecutive patients who were attending one of two endoscopy centres in Christchurch, New Zealand, for colonoscopy and a subgroup of participants from the general public who did not undergo colonoscopy. Participants with known GI disease other than an FGID were excluded. Those with symptoms were recruited as cases, whilst those without symptoms were recruited as controls. In the days prior to preparation for colonoscopy, or an agreeable time for those not undergoing colonoscopy, demographic, symptom, psychological, dietary, and health data were collected in addition to biological samples (breath, faeces, blood, and urine). Colonic biopsies were taken at the time of colonoscopy from participants in the colonoscopy subgroup. Results: Between July 2016 and December 2018, 349 participants were recruited, 315 of whom completed the study, 220 participants were from the colonoscopy subgroup, and 95 from the non-colonoscopy subgroup. This included 129 controls and 186 cases (57 IBS-diarrhoea predominant, 30 IBS-constipation predominant, 41 IBS-mixed, 42 FC, and 16 FD). The mean age of FGID cases was 53.4 years and controls 54.4 years. Cases (149/186, 80.1%) and controls (57/72, 55.8%) were predominantly female. Education levels were similar across the cohort. Smoking and alcohol rates were also similar. Biological samples were collected as planned from participants. Conclusions: The COMFORT cohort is a unique clinical cohort of FGID cases and controls with a wide range of demographic, dietary, clinical, psychological, and health data in addition to biological samples. Future research will aim to use a systems biology approach to establish the potential role of diet, host-microbiome interactions, and other factors in the pathogenesis of FGIDs.

Gene expression and phytohormone contents were measured in response to elevating ascorbate in the absence of other confounding stimuli such as high light and abiotic stresses. Young Arabidopsis plants were treated with 25 mM solutions of l-galactose pathway intermediates l-galactose (l-gal) or l-galactono-1,4-lactone (l-galL), as well as L-ascorbic acid (AsA), with 25 mM glucose used as control. Feeding increased rosette AsA 2- to 4-fold but there was little change in AsA biosynthetic gene transcripts. Of the ascorbate recycling genes, only Dehydroascorbate reductase 1 expression was increased. Some known regulatory genes displayed increased expression and included ANAC019, ANAC072, ATHB12, ZAT10 and ZAT12. Investigation of the ANAC019/ANAC072/ATHB12 gene regulatory network revealed a high proportion of ABA regulated genes. Measurement of a subset of jasmonate, ABA, auxin (IAA) and salicylic acid compounds revealed consistent increases in ABA (up to 4.2-fold) and phaseic acid (PA, up to 5-fold), and less consistently certain jasmonates, IAA, but no change in salicylic acid levels. Increased ABA is likely due to increased transcripts for the ABA biosynthetic gene NCED3. There were also smaller increases in transcripts for transcription factors ATHB7, ERD1, and ABF3. These results provide insights into how increasing AsA content can mediate increased abiotic stress tolerance.

Kiwifruit (Actinidia chinensis) is a dioecious, long-living woody perennial vine. Reduced generation time and induction of hermaphroditism can accelerate crop improvement and facilitate alternative farming for better food security in the face of climate change. Previous studies identified that CENTRORADIALIS genes CEN and CEN4 act to repress flowering, whilst the male-specific Shy Girl (SyGl) gene with homology to type-C cytokinin response regulators could repress gynoecium development in model plants. Here we use CRISPR/Cas9 to mutagenize CEN, CEN4 and SyGl in the male kiwifruit A. chinensis 'Bruce'. Biallelic mutations of CEN and CEN4 generated rapid-flowering male plants, and simultaneous targeting of CEN4 and SyGl gave rise to rapid-flowering hermaphrodites with restored gynoecial function and viable pollen, providing functional evidence for the role of SyGl in suppression of feminization. Analysis of ovary tissues identified genes that contribute to carpel development and revealed that SyGl affected both cytokinin profiles and the expression of genes involved in cytokinin metabolism and signalling. The plant lines generated by CEN4/SyGl knockout could self-pollinate and produce fast-flowering offspring. These results establish that SyGI acts as the suppressor of feminization in kiwifruit and demonstrate the potential for accelerated breeding in an outcrossing horticultural woody perennial.

Epidemics of obesity and type 2 diabetes drive strong consumer interest in plant-based low-calorie sweeteners. Trilobatin is a sweetener found at high concentrations in the leaves of a range of crabapple (Malus) species, but not in domesticated apple (Malus × domestica) leaves, which contain trilobatin’s bitter positional isomer phloridzin. Variation in trilobatin content was mapped to the Trilobatin locus on LG 7 in a segregating population developed from a cross between domesticated apples and crabapples. Phloretin glycosyltransferase2 (PGT2) was identified by activity-directed protein purification and differential gene expression analysis in samples high in trilobatin but low in phloridzin. Markers developed for PGT2 cosegregated strictly with the Trilobatin locus. Biochemical analysis showed PGT2 efficiently catalyzed 4′-o-glycosylation of phloretin to trilobatin as well as 3-hydroxyphloretin to sieboldin. Transient expression of double bond reductase, chalcone synthase, and PGT2 genes reconstituted the apple pathway for trilobatin production in Nicotiana benthamiana. Transgenic M. × domestica plants overexpressing PGT2 produced high concentrations of trilobatin in young leaves. Transgenic plants were phenotypically normal, and no differences in disease susceptibility were observed compared to wild-type plants grown under simulated field conditions. Sensory analysis indicated that apple leaf teas from PGT2 transgenics were readily discriminated from control leaf teas and were perceived as significantly sweeter. Identification of PGT2 allows marker-aided selection to be developed to breed apples containing trilobatin, and for high amounts of this natural low-calorie sweetener to be produced via biopharming and metabolic engineering in yeast.

DORMANCY-ASSOCIATED MADS-box (DAM) and SHORT VEGETATIVE PHASE (SVP) genes have been implicated in the regulation of winter dormancy in perennials. Ectopic expression of apple (Malus × domestica Borkh. ‘Royal Gala’) DAM and SVP genes delays budbreak and constrains lateral shoot outgrowth. In this study, we used RNA interference (RNAi) to simultaneously target all apple DAM and SVP genes in order to study their role and mode of action in the regulation of bud dormancy, budbreak and flowering. A synthetic construct carrying a hairpin fragment assembled from sequences specific to coding regions of three DAM and two SVP genes was used to generate transgenic lines. Reduced expression of DAM/SVP genes resulted in delayed leaf senescence and abscission in autumn, failure to enter bud dormancy in winter and continual growth of new leaves regardless of the season for over 3 years. Precocious flowering but normal flower morphology, fertility and fruit development were observed. The non-dormant phenotype was associated with modified phytohormone composition. The content of gibberellins (GAs) and jasmonates (JAs) was significantly increased in terminal buds of RNAi lines compared with wildtype plants, accompanied by elevated expression of the key GA biosynthesis pathway gene GIBBERELLIN 20 OXIDASE-2 (MdGA20ox-2) along with the FLOWERING LOCUS T gene MdFT2. The key mediator of plasmodesmatal closure, MdCALLOSE SYNTHASE 1 (MdCALS1), was repressed in RNAi lines. This study provides functional evidence for the role of DAM/SVP genes in vegetative phenology of apple and paves the way for production of low-chill varieties suitable for growth in warming climates.

Bioactive compounds such as anthocyanins, proanthocyanins and other polyphenols are found in a wide variety of fruits and vegetables, and consumption of these have been associated with reduced lung inflammation and improved lung function in asthma and other lung diseases. We investigated whether a combination of Boysenberry and apple juice, found in BerriQi® Boysenberry and apple juice concentrate, could reduce the allergic airways inflammation associated with asthma. We characterised the polyphenolic components in BerriQi® Boysenberry and apple juice concentrate and identified the main compounds as cyanidin glycosides, ellagitannins, and chlorogenic acid. We found that consumption of 2.5 mg/kg of total anthocyanins from the BerriQi® Boysenberry and apple juice concentrate significantly reduced eosinophil infiltration following acute ovalbumin (OVA) exposure in a mouse model of allergic airways inflammation. We found that BerriQi® Boysenberry and apple juice concentrate consumption increased M2 (CD206+) macrophages and the production of the M2-associated cytokines CXCL10 and CCL4 within the lung. These results suggest that consumption of BerriQi® Boysenberry and apple juice concentrate promotes a shift towards an anti-inflammatory environment within the lung leading to reduced immune cell infiltration and tissue damage.

Following cell division, fruit growth is characterized by both expansion through increases in cell volume and biomass accumulation in cells. Fruit growth is limited by carbon starvation; however, the mechanism controlling fruit growth under restricted carbohydrate supply is poorly understood. In a previous study using red-fleshed kiwifruit, we showed that long-term carbon starvation had detrimental effects on carbohydrate, anthocyanin metabolism, and fruit growth. To elucidate the mechanisms underlying the reduction in fruit growth during kiwifruit development, we integrated phytohormone profiling with transcriptomic and developmental datasets for fruit under high or low carbohydrate supplies. Phytohormone profiling of the outer pericarp tissue of kiwifruit showed a 6-fold reduction in total cytokinin concentrations in carbon-starved fruit, whilst other hormones were less affected. Principal component analysis visualised that cytokinin composition was distinct between fruit at 16 weeks after mid bloom, based on their carbohydrate supply status. Cytokinin biosynthetic genes (IPT, CYP735A) were significantly downregulated under carbon starvation, in agreement with the metabolite data. Several genes that code for expansins, proteins involved in cell wall loosening, were also downregulated under carbon starvation. In contrast to other fleshy fruits, our results suggest that cytokinins not only promote cell division, but also drive fruit cell expansion and growth in kiwifruit.

Introduction of Fourier-transform infrared (FTIR) spectroscopy would enable breeders to screen phenotypic variability in multiple fruit from large numbers of progeny. Thus far, however, there has been no comprehensive attempt to develop chemometric models for determination of quality attributes of small berry species by this approach. FTIR spectra (1800–900 cm−1) of juice from breeding populations of four genera (Actinidia, specifically red kiwifruit, Ribes, Rubus and Vaccinium) were analysed by partial least squares regression to determine the possibility of measuring soluble solids (SS), titratable acidity (TA) and total anthocyanin (ACY) concentration simultaneously using global prediction models. SS, TA and ACY concentrations across all berry juices ranged between 4.1 and 22.4 °Brix, 0.1–5.5% citric acid and 2–4697 ppm, respectively. R2 (coefficient of determination in cross-validation) and SECV (standard error of cross-validation) statistics for global models were 0.996 (0.22 °Brix), 0.996 (0.08% citric acid) and 0.893 (280 ppm). Analysis of data sets for individual berry types separately demonstrated that it was possible to develop models with superior prediction statistics for each attribute. However, these were not necessarily robust when validated against data from different seasons, locations or breeding selections. These global models represent an advance for researchers wishing to screen substantial fruit populations more rapidly.

Plant metabolomics within field-based food production systems is challenging owing to environmental variability and the complex architecture and metabolic growth cycles of plants. Kiwifruit cultivars of Actinidia chinensis are vigorous perennial vines grown as clones in highly structured orchard environments, intensively managed to maximize fruit yield and quality. To understand the metabolic responses of vines to orchard management practices, we needed to better understand the various sources of metabolic variability encountered in the orchard. Triplicate composite leaf, internode and fruit (mature and immature) samples were collected from each of six Actinidia chinensis var. deliciosa ‘Hayward’ and A. chinensis var. chinensis ‘Zesy002’ kiwifruit vines at three times during the growing season and measured by LC-MS. In general, there was more variation in metabolite concentrations within vines than between vines, with ‘Hayward’ showing a greater percentage of within-vine variability than ‘Zesy002’ (c. 90 vs. 70% respectively). In specific tissues, the sampler, infection by Pseudomonas syringae var. actinidiae and the rootstock also influenced metabolite variability. A similar pattern of metabolic variability was observed from quantitative analysis of specific carbohydrates and phytohormones. High within-vine metabolic variability indicates that it is more important to obtain sufficient replicate samples than to sample from multiple vines. These data provide an objective basis for optimizing metabolite sampling strategies within kiwifruit orchards.

Kiwifruit may be stored for prolonged periods at low temperatures during which time the fruit continue to ripen slowly and dehydration occurs. Abscisic acid (ABA) plays important roles in both fruit ripening and environmental stress response, including dehydration. To investigate the inter-relationships among postharvest dehydration, ethylene production, ABA and fruit ripening, two independent experiments were undertaken with ‘Hayward’ kiwifruit. Water loss was manipulated by holding fruit in three environments of different humidities at 20 °C for up to 6 d, and at 0 °C for up to 16 weeks. The impacts of the treatments over time after harvest were investigated on fruit softening, ABA content and ethylene production, and on the expression of genes of the ABA biosynthetic pathway and ethylene metabolism. Water loss induced earlier ethylene production and accelerated softening late in storage. However, these changes were not directly related to increased ABA content. The strongest association seen was between tissue ABA content and fruit firmness, irrespective of the degree of water loss. In conclusion, it is suggested that the accelerated softening of ‘Hayward’ kiwifruit caused by dehydration may be mediated via ethylene, but that the precise role for ABA, either directly or indirectly, is as yet difficult to ascribe.

Consumer acceptance of fruit is determined by size, flavour and ripeness. In this study we investigated how altering the carbohydrate supply to Actinidia chinensis var. chinensis ‘Zesy002’ kiwifruit altered the balance between growth and accumulation of metabolites. Canes were phloem girdled and fruit thinned to a leaf-to-fruit ratio (L:F) of either 2 (Low carbohydrate) or 6 (High carbohydrate) at either 38 (Early) or 86 (Late) days after anthesis (DAA) and compared with ungirdled control canes with a L:F of 3. Fruit growth, metabolite accumulation, cytokinin concentrations and maturation were monitored and the sensory attributes of ripe fruit were assessed. The final weight of Early-High and Late-High carbohydrate fruit was 38% and 16% greater compared with control fruit. High carbohydrate fruit had increased starch, soluble sugar and cytokinin concentrations and fruit began to mature earlier and those with a Low carbohydrate had decreased concentrations and matured later compared with control fruit. Control fruit were described by consumers as more acidic and under-ripe compared with those from Early-High carbohydrate canes, but as sweeter than those from Low carbohydrate canes. This study showed that carbohydrate supply can have a major impact on the growth, sugar accumulation and maturity of ‘Zesy002’ fruit sinks.

Blanching is an important process in the preparation of navy beans (Phaseolus vulgaris L.) for canning. We here explore the effect of blanching which can profoundly affect protein composition and introduce protein-primary-level modifications. Amino acid analysis showed significantly decreased protein abundance (58.5%) in blanched beans compared to raw beans. Proteomic analyses revealed a decrease in high molecular weight isoforms of the major storage globulin proteins phaseolin (mean fold-change −3.7) and legumin (mean fold-change −2.5) and concomitant increase in their low molecular weight isoforms (mean fold-change 6.4 and 8.3, respectively). Blanched beans also had decreased abundance of lipoxygenase (mean fold-change −13.1), an enzyme responsible for product spoilage during storage. Increased lysinoalanine (up to 47%) and highly modified protein fragments were found in the processing waters, indicating heat- induced modifications. Correlating these molecular level changes thus provides a basis for evaluating how processing parameters can be modified to increase protein food quality.

Intermittent fasting improves metabolic and cardiac health. However, increased hunger towards the end of the fasting period may affect compliance and limit its application. Our aim was to determine the effect of anorexigenic agent co-therapy on subjective ratings of appetite during the 16&ndash, 24 h period of a day-long water-only intermittent fast. Thirty adult men were recruited and required to fast for 24 h from 18:00 h to 18:00 h on the same day of the week for three subsequent weeks. Treatments of either a placebo or one of two doses (high dose, HD: 250 mg or low dose, LD: 100 mg) of a bitter hops-based appetite suppressant (Amarasate&reg, ) were given twice per day at 16 and 20 h into the fast. From 18&ndash, 24 h of the 24 h fast, both the HD and LD treatment groups exhibited a statistically significant (p &lt, 0.05) &gt, 10% reduction in hunger. Additionally, the expected lunchtime increase in hunger that was present in the placebo group (12:00 h) was absent in both the HD and LD groups. These data suggest that appetite suppressant co-therapy may be useful in reducing hunger during intermittent fasting, and show that bitter compounds may regulate appetite independently of meal timing.

OBJECTIVES: The links between food, gastrointestinal (GI) function and comfort, and mental well-being are at the forefront of nutritional research. Irritable Bowel Syndrome (IBS) is a functional GI disorder and the combination of detailed patient reported outcomes (PROs) combined with omic data could better define these disorders. METHODS: The Christchurch COMFORT cohort is a case-control study: 337 participants with functional GI disorders (functional constipation FC, functional diarrhoea FD, IBS constipation IBS-C, IBS diarrhoea IBS-D) and asymptomatic controls. Demographics, symptom scores, psychological scores and dietary intake were recorded using the ‘Modified Hunter New England’ (Rome IV diagnostic criteria questionnaires; Medical History Questions; Hospital Anxiety and Depression Scale, HADS). Demographics included Structured Assessment of Gastrointestinal Symptoms (SAGIS); and Diet Diary and Live Symptoms Score (FAST) along with PRO Measurement Information System (PROMIS). Biological samples collected for an untargeted LCMS analysis of plasma and shot-gun metagenome analysis of faecal DNA. Ethical approval was obtained from the University of Otago Human Ethics Committee (H16/094). RESULTS: Symptom questionnaires were able to cluster 287 subjects into: IBS (42%), functional disorders (18%) and healthy controls (40%). Within IBS, 46% were IBS-D, 23% were IBS-C and 31% were IBS-M. Severity scores were higher for all IBS cases with PROMIS-GI scales. A higher score of health worry was reported in IBS-C than other IBS subtypes. HADS anxiety and depression scales were higher in IBS cases (vs. healthy controls). The FAST diary symptoms correlated with PROMIS GI scales (exception for constipation). Metabolomic analyses detected differential plasma metabolites and pathways (bile acids, lipids, specific amino acids) affected between IBS-C + FC vs IBS-D + FD, healthy vs IBS-D and healthy vs IBS-C. Metagenomics suggests that carbohydrate, methane, and sulfur metabolism may be important in IBS. CONCLUSIONS: These data will allow us to apply a systems biology approach to identify key pathways and correlate them with the questionnaire data to better understand functional GI disorders. FUNDING SOURCES: Funded by the NZ National Science Challenge High-Value Nutrition programme.

Background Affective responses experienced during exercise are a significant determinant on exercise adherence. We have previously demonstrated that consumption of New Zealand (NZ) blackcurrants preserves cognition by attenuating the feeling of fatigue. This positive affective response correlated with the ability of blackcurrant polyphenols to support monoamine neurotransmission via inhibition of monoamine oxidase-B (MAO-B) activity. Here we explore how the consumption of a NZ blackcurrant juice (BJ) influenced affective responses and potential ergogenic action on the motivation to adhere to a low impact walking exercise. Methods In a parallel randomized controlled study (Trial registration #: ACTRN12617000319370p, registered 28th February 2017, http://www.anzctr.org.au/), 40 healthy sedentary male and female participants drank a BJ or matched placebo (PLA) (n = 20 per group), 1 h prior to a self-motivated treadmill walk, where heart rate and affective responses (exertion [ES] or feeling / mood [FS]) scores) were recorded at 3 or 5 min intervals. Blood glucose, lactate, malondialdehyde (MDA) and platelet MAO-B activity were measured pre- and post-exercise and comparisons were conducted using with Student’s t-tests. Subjective data were analysed using 2-way ANOVA with appropriate post hoc tests. Results Consuming a BJ 1 h prior to exercise caused a 90% decline in platelet MAO-B activity. The exercise had no significant (p > 0.05) effect on blood lactate, glucose or plasma MDA levels. Assessment of affective responses over the first 60 mins (adjusting for participant drop-out) revealed a time-dependent ES increase in both groups, with ES reported by participants in the BJ group consistently lower than those in the PLA group (p 10 km compared to only 10% for the PLA group (p = 0.28), statistical significance was not achieved. Conclusion Our findings demonstrate that drinking a polyphenolic-rich NZ blackcurrant juice 1 h prior to exercise supports positive affective responses during a self-motivated exercise.

Anthocyanins are a group of secondary metabolites that colour fruit and flowers orange, red, purple or blue depending on a number of factors, such as the basic structure, co-pigmentation, metal ion complexation and vacuolar pH. The biosynthesis of anthocyanin is regulated at the transcriptional level by a group of transcription factors, the MYB–bHLH–WD40 (MBW) complex. In this study, the purple colouration in several kiwifruit (Actinidia) species was identified and characterised as red cyanidin-based and blue delphinidin-based anthocyanins. The differential pigmentation in the skin and flesh can be attributed to the differential ratio of cyanidin and delphinidin derivatives accumulated in the total anthocyanin profile. The expression of anthocyanin biosynthetic genes chalcone synthase (CHS), flavonoid 3-O-glucosyltransferase (F3GT), flavonoid 3′-hydroxylase (F3′H) and flavonoid 3′5′-hydroxylase (F3′5′H) is crucial for anthocyanin accumulation. However, the balance of expression of the F3′H and F3′5′H genes appears responsible for the ratio of cyanidin and delphinidin derivatives, while a lack of CHS, F3GT and MYB110 expression is responsible for a lack of total anthocyanins. The transcriptional regulation of the F3′H and F3′5′H promoters by the R2R3 MYB transcription factor MYB110 is markedly different in tobacco transient assays. When kiwifruit MYB10 or MYB110 are over-expressed in Actinidia chinensis both cyanidin-based and delphinidin-based anthocyanins are elevated, but F3′H and F3′5′H genes are not strongly correlated with MYB expression. These results suggest that the core kiwifruit anthocyanin pathway genes are dependent on characterised MYB transcription factors, while other regulatory proteins are more directly responsible for the expression of the F3′H and F3′5′H genes., Anthocyanins: Coloring Kiwifruit The genetic processes that determine the colors in purple kiwifruit have been revealed by researchers in New Zealand. Anthocyanin pigments are not only essential for plant growth and development, but also provide color to make fruit attractive to pollinators and indicate ripeness for the food industry. Andrew Allan and co-workers at the University of Auckland and The New Zealand Institute for Plant and Food Research Limited examined several different kiwifruit species, and found that their color was determined by the ratio of red cyanidin-based to blue delphinidin-based anthocyanins. The team identified four biosynthetic genes that are crucial for accumulating anthocyanins, and showed that two of those genes control the red-to-blue pigment ratio. This new understanding of the anthocyanin profile could allow metabolic engineering of novel fruit colors, possibly with greater health benefits for humans.

Summary Apple (Malus × domestica) accumulates bioactive ursane-, oleanane-, and lupane-type triterpenes in its fruit cuticle, but their biosynthetic pathway is still poorly understood. We used a homology-based approach to identify and functionally characterize two new oxidosqualene cyclases (MdOSC4 and MdOSC5) and one cytochrome P450 (CYP716A175). The gene expression patterns of these enzymes and of previously described oxidosqualene cyclases were further studied in 20 apple cultivars with contrasting triterpene profiles. MdOSC4 encodes a multifunctional oxidosqualene cyclase producing an oleanane-type triterpene, putatively identified as germanicol, as well as β-amyrin and lupeol, in the proportion 82 : 14 : 4. MdOSC5 cyclizes 2,3-oxidosqualene into lupeol and β-amyrin at a ratio of 95 : 5. CYP716A175 catalyses the C-28 oxidation of α-amyrin, β-amyrin, lupeol and germanicol, producing ursolic acid, oleanolic acid, betulinic acid, and putatively morolic acid. The gene expression of MdOSC1 was linked to the concentrations of ursolic and oleanolic acid, whereas the expression of MdOSC5 was correlated with the concentrations of betulinic acid and its caffeate derivatives. Two new multifuntional triterpene synthases as well as a multifunctional triterpene C-28 oxidase were identified in Malus × domestica. This study also suggests that MdOSC1 and MdOSC5 are key genes in apple fruit triterpene biosynthesis.

The aim of this study was to provide insight into how curcumin reduces colon inflammation in the Mdr1a(-/-) mouse model of human inflammatory bowel disease using a combined transcriptomics and proteomics approach. Mdr1a(-/-) and FVB control mice were randomly assigned to an AIN-76A (control) diet or AIN-76A+0.2% curcumin. At 21 or 24weeks of age, colonic histological injury score (HIS) was determined, colon mRNA transcript levels were assessed using microarrays and colon protein expression was measured using 2D gel electrophoresis and LCMS protein identification. Colonic HIS of Mdr1a(-/-) mice fed the AIN-76A diet was higher (P

Aim: To evaluate blackcurrant anthocyanin-rich extract (BAE) consumption on time- and dose-dependent plasma anthocyanin bioavailability and conduct a pilot study to explore the potential effect of BAE in promoting recovery from exercise-induced oxidative stress, and maintenance of circulating neutrophil function.Methods: Time- and dose-dependent blackcurrant anthocyanin bioavailability was assessed using LC-MS in 12 participants over 6 h after the ingestion of a placebo or BAE containing 0.8, 1.6, or 3.2 mg/kg total anthocyanins. In a separate pilot intervention exercise trial, 32 participants consumed either a placebo or 0.8, 1.6, or 3.2 mg/kg BAE (8 individuals per group), and then 1 h later performed a 30 min row at 70% VO2max. Blood was collected during the trial for oxidative, antioxidant, inflammatory, and circulating neutrophil status.Results: Consumption of BAE caused a time- and dose-dependent increase in plasma anthocyanins, peaking at 2 h after ingestion of 3.2 mg/kg BAE (217 ± 69 nM). BAE consumed 1 h prior to a 30 min row had no effect on plasma antioxidant status but hastened the recovery from exercise-induced oxidative stress: By 2 h recovery, consumption of 1.6 mg/kg BAE prior to exercise caused a significant (P < 0.05) 34 and 32% decrease in post-exercise plasma oxidative capacity and protein carbonyl levels, respectively, compared to placebo. BAE consumption prior to exercise dose-dependently attenuated a small, yet significant (P < 0.01) transient 13 ± 2% decline in circulating neutrophils observed in the placebo group immediately post-exercise. Furthermore, the timed consumption of either 1.6 or 3.2 mg/kg BAE attenuated a 17 ± 2.4% (P < 0.05) decline in neutrophil phagocytic capability of opsonised FITC-Escherichia coli observed 6 h post-exercise in the placebo group. Similarly, a dose-dependent increase in neutrophil surface expression of complement receptor-3 complex (CR3, critical for effective phagocytosis of opsonised microbes), was observed 6 h post-exercise in both 1.6 and 3.2 mg/kg BAE intervention groups.Conclusions: Consumption of BAE (>1.6 mg/kg) 1 h prior to exercise facilitated recovery from exercise-induced oxidative stress and preserved circulating neutrophil function. This study provides data to underpin a larger study designed to evaluate the efficacy of timed BAE consumption on post-exercise recovery and innate immunity.

Emerging evidence from human intervention trials indicates health benefits of consuming blackcurrant fruit, including improvements to cognitive performance, modulation of blood flow, regulation of blood glucose and inhibition of enzymes underpinning normal cognitive function. Of particular relevance is our previous demonstration of monoamine oxidase (MAO)-A and B inhibition after the consumption of a New Zealand “Blackadder” blackcurrant juice in humans. The current study uses a double-blind, placebo-controlled, randomised cross- over design to assess the pharmacodynamics of the effects on platelet MAO-B inhibition and associated substrates, plasma prolactin levels and blood glucose levels after consumption of a single serve of “Blackadder” blackcurrant juice standardised to 500 mg polyphenols. Eight healthy male (20-–35 years) participants completed the trial. Measurements were obtained at baseline 15, 30, 45, 60, 100, 120, 150, 180, 240 mins and 24 h post dose. A fast, absolute and reversible inhibition of blood platelet MAO-B (P < 0.001) and a significant but delayed reduction in plasma prolactin (P < 0.001) were observed following the consumption of “Blackadder” blackcurrant juice when compared to a placebo control. No interpretable changes in substrates of MAO or associated metabolites were seen. These data provide a clear time course of the reversible inhibition of MAO-B after the single consumption of a of New Zealand “Blackadder” blackcurrant juice standardised at 500 mg of polyphenols and, therefore, provide a therapeutic window on which to base future nutritional interventions.

The consumption of berry fruits engenders a number of benefits in animal models, including improvements in cognitive performance, slowing of cognitive decline during natural ageing, and neuroprotection. These findings, along with limited human epidemiological evidence, suggest a potential role for the consumption of berry fruit polyphenols in improving human cognitive performance. The current study assessed the effects of two blackcurrant extracts on cognitive outcomes, mood, autonomic measures, peripheral and central monoamine tone, and anthocyanin bioavailability to plasma. A randomised, double-blind, placebo-controlled, crossover study was conducted using 36 healthy young participants (18–35 years). Findings from the intervention illustrate a cognitive benefit of acute blackcurrant supplementation in healthy young humans and the first description of a clinically significant inhibition of monoamine oxidase-B and monoamine oxidase-A using a commonly consumed fruit. These data also illustrate that compounds other than anthocyanins may be responsible for the observed in vivo MAO inhibition and that the degree of processing and the cultivar of blackcurrant fruit used substantially alter the neuroendocrinological and cognitive benefits conveyed.

Pseudomonas syringae pv. actinidiae (Psa) and Pseudomonas syringae pv. actinidifoliorum (Pfm) are closely related pathovars infecting kiwifruit, but Psa is considered one of the most important global pathogens, whereas Pfm is not. In this study of Actinidia deliciosa ‘Hayward’ responses to the two pathovars, the objective was to test whether differences in plant defense responses mounted against the two pathovars correlated with the contrasting severity of the symptoms caused by them. Results showed that Psa infections were always more severe than Pfm infections, and were associated with highly localized, differential expression of phytohormones and putative defense gene transcripts in stem tissue closest to the inoculation site. Phytohormone concentrations of jasmonic acid (JA), jasmonate isoleucine (JA-Ile), salicylic acid (SA) and abscisic acid were always greater in stem tissue than in leaves, and leaf phytohormones were not affected by pathogen inoculation. Pfm inoculation induced a threefold increase in SA in stems relative to Psa inoculation, and a smaller 1.6-fold induction of JA. Transcript expression showed no effect of inoculation in leaves, but Pfm inoculation resulted in the greatest elevation of the SA marker genes, PR1 and glucan endo-1,3-beta-glucosidase (β-1,3-glucosidase) (32- and 25-fold increases, respectively) in stem tissue surrounding the inoculation site. Pfm inoculation also produced a stronger response than Psa inoculation in localized stem tissue for the SA marker gene PR6, jasmonoyl-isoleucine-12-hydrolase (JIH1), which acts as a negative marker of the JA pathway, and APETALA2/Ethylene response factor 2 transcription factor (AP2 ERF2), which is involved in JA/SA crosstalk. WRKY40 transcription factor (a SA marker) was induced equally in stems by wounding (mock inoculation) and pathovar inoculation. Taken together, these results suggest that the host appears to mount a stronger, localized, SA-based defense response to Pfm than Psa.

Interaction Between Strigolactone and Cytokinin on Axillary and Adventitious Bud Development in Zantedeschia Strigolactones are known to be involved in the control of branching, particularly axillary bud outgrowth. However, the effect of strigolactone on adventitious bud development is presently unknown, and the interaction of strigolactone with cytokinin on bud outgrowth has not been fully understood. Using a germination assay, high strigolactone levels were detected in a low-branched cultivar (Best Gold) but was less abundant in a high-branched cultivar (Goldilocks), particularly at an early stage of the growth cycle when branching was not visibly evident. In contrast, the concentration of cytokinins directly correlated with branching, suggesting an antagonistic interaction between strigolactones and cytokinins on branching. Supporting this hypothesis, strigolactone reduced the cytokinin stimulation of both axillary and adventitious shoot number in Zantedeschia grown in vitro. The reduced cytokinin concentration in a phenotypically low branched cultivar may have been associated with the inhibition effect of strigolactones. Hence, the fact that the alteration of branching in Zantedeschia or other horticultural species depends on the hormonal balance between these two hormones is quite likely. The idea may be useful for the generation of plants of desired branching at least within the in vitro system.

Summary The polyphenol profile of apple (Malus x domestica) is dominated by the dihydrochalcone glycoside phloridzin, however, its physiological role is yet to be elucidated. Biosynthesis of phloridzin occurs as a side-branch of the main phenylpropanoid pathway with the final step mediated by the phloretin-specific glycosyltransferase UGT88F1. Unexpectedly, given that UGTs are sometimes viewed as ‘decorating enzymes’, UGT88F1 knockdown lines were severely dwarfed with greatly reduced internode lengths, narrow lanceolate leaves, and changes in leaf and fruit cellular morphology. These changes suggested that auxin transport had been altered in the knockdown lines, which was confirmed in assays showing that auxin flux from the shoot apex was increased in the transgenic lines. Metabolite analysis revealed no accumulation of the phloretin aglycone, as well as decreases in many non-target phenylpropanoid compounds. This decreased accumulation of metabolites appeared to be mediated by repression of the phenylpropanoid pathway via reduction in key transcript levels (e.g. phenylalanine ammonia lyase, PAL) and enzyme activities (PAL and chalcone synthase). Application of exogenous phloridzin to the UGT88F1 knockdown lines in tissue culture enhanced axial leaf growth and partially restored some aspects of ‘normal’ apple leaf growth. Together, our results strongly implicate dihydrochalcones as critical compounds in modulating phenylpropanoid pathway flux and establishing auxin patterning early in apple development. This article is protected by copyright. All rights reserved.

Heteroxylans in the plant cell wall have been proposed to have a role analogous to that of xyloglucans or heteromannans, forming growth-restraining networks by interlocking cellulose microfibrils. A xylan endotransglycosylase has been identified that can transglycosylate heteroxylan polysaccharides in the presence of xylan-derived oligosaccharides. High activity was detected in ripe fruit of papaya (Carica papaya), but activity was also found in a range of other fruits, imbibed seeds and rapidly growing seedlings of cereals. Xylan endotransglycosylase from ripe papaya fruit used a range of heteroxylans, such as wheat arabinoxylan, birchwood glucuronoxylan and various heteroxylans from dicotyledonous primary cell walls purified from tomato and papaya fruit, as donor molecules. As acceptor molecules, the enzyme preferentially used xylopentaitol over xylohexaitol or shorter-length acceptors. Xylan endotransglycosylase was active over a broad pH range and could perform transglycosylation reactions up to 55 °C. Xylan endotransglycosylase activity was purified from ripe papaya fruit by ultrafiltration and cation exchange chromatography. Highest endotransglycosylase activity was identified in fractions that also contained high xylan hydrolase activity and correlated with the presence of the endoxylanase CpaEXY1. Recombinant CpaEXY1 protein transiently over-expressed in Nicotiana benthamiana leaves showed both endoxylanase and xylan endotransglycosylase activities in vitro, suggesting that CpaEXY1 is a single enzyme with dual activity in planta. Purified native CpaEXY1 showed two- to fourfold higher endoxylanase than endotransglycosylase activity, suggesting that CpaEXY1 may act primarily as a hydrolase. We propose that xylan endotransglycosylase activity (like xyloglucan and mannan endotransglycosylase activities) could be involved in remodelling or re-arrangement of heteroxylans of the cellulose-non-cellulosic cell wall framework.

SummaryA continuous supply of boron (B) is vital for the formation of new cells, with reproductive growth being particularly sensitive to boron deficiency. An inadequate B supply therefore has the potential to effect fruit set, which is a major problem in avocado (Persea americana Mill.) cultivation. B is not phloem mobile in many plant species, but when sorbitol or mannitol (both polyols) are present in phloem sap, then B forms a complex with the polyol, enabling transport in the phloem. Phloem mobility of B has never been demonstrated in avocado. Avocados are unusual in that they produce the 7-carbon sugar, D-mannoheptulose, and its polyol form, perseitol. In this work, we demonstrate that B forms a complex with perseitol in vitro, that both B and perseitol are found in avocado phloem sap, that B is found in higher concentrations within immature leaves than in mature leaves, and that 10B is exported from mature leaves into inflorescences and new leaves. All these findings are consistent with B being tra...

Inflammatory bowel disease (IBD) is characterized by intestinal inflammation and is believed to involve complex interactions between genetic, immunological, and environmental factors. We measured changes in the proteome associated with bacterially induced intestinal inflammation in the interleukin 10 gene-deficient (Il10(-/-)) mouse model of IBD, established effects of the dietary polyunsaturated fatty acids (PUFAs) n-3 eicosapentaenoic acid (EPA) and n-6 arachidonic acid (AA) on protein expression (using oleic acid as a control fatty acid), and compared these changes with previously observed transcriptome changes in the same model. Ingenuity pathways analysis of proteomics data showed bacterially induced inflammation was associated with reduced expression of proteins from pathways of metabolism and digestion/absorption/excretion of nutrients/ions, and increased expression of cellular stress and immune response proteins. Both PUFA treatments showed anti-inflammatory activity; EPA appeared to act via the PPARα pathway, whereas AA appeared to increase energy metabolism and cytoskeletal organization and reduce cellular stress responses, possibly enabling a more robust response to inflammation. While there was agreement between proteomic and transcriptomic data with respect to pathways, there was limited concordance between individual gene and protein data, reflecting the importance of having both gene and protein data to better understand complex diseases such as IBD.

Inflammatory bowel disease (IBD) is a collective term for conditions characterised by chronic inflammation of the gastrointestinal tract involving an inappropriate immune response to commensal micro-organisms in a genetically susceptible host. Previously, aqueous and ethyl acetate extracts of gold kiwifruit (Actinidia chinensis) or green kiwifruit (A. deliciosa) have demonstrated anti-inflammatory activity usingin vitromodels of IBD. The present study examined whether these kiwifruit extracts (KFE) had immune-modulating effectsin vivoagainst inflammatory processes that are known to be increased in patients with IBD. KFE were used as a dietary intervention in IL-10-gene-deficient (Il10− / −) mice (anin vivomodel of IBD) and the C57BL/6J background strain in a 3 × 2 factorial design. While allIl10− / −mice developed significant colonic inflammation compared with C57BL/6J mice, this was not affected by the inclusion of KFE in the diet. These findings are in direct contrast to our previous study where KFE reduced inflammatory signalling in primary cells isolated fromIl10− / −and C57BL/6J mice. Whole-genome gene and protein expression level profiling indicated that KFE influenced immune signalling pathways and metabolic processes within the colonic tissue; however, the effects were subtle. In particular, expression levels across gene sets related to adaptive immune pathways were significantly reduced using three of the four KFE in C57BL/6J mice. The present study highlights the importance of investigating food components identified by cell-based assays with appropriatein vivomodels before making dietary recommendations, as a food that looks promisingin vitromay not be effectivein vivo.

The positive roles of fruit polyphenols (PPs) and dietary fibres (DFs) in promoting human health and well-being are now well recognised. This study investigates the effects of added fruit PPs and pectin on the composition and extractability of PPs and proteins from finished breads, and complements our recently published papers that focus on the rheological and physicochemical properties of breads containing added PPs and pectin. Breads were prepared in the absence or presence of a high methoxyl (HM) apple pectin and kiwifruit, blackcurrant or apple PP extract, and subjected to texture measurements (crumb firmness), informal sensory evaluations and chemical analyses. An epicatechin standard was also used in place of the fruit PP extract to estimate the extractability and recovery of PPs from bread. Results show that adding a PP extract and pectin to bread modified bread firmness, specific volume, recovery of added PPs, amount of free thiol groups of bread proteins, and also the extractability of PPs and proteins from bread. The recovery of added PPs was the lowest for the APE + HM pectin bread which had the highest bread firmness. High performance liquid chromatography (HPLC)-mass spectrometry (MS) PP profiling revealed that baking caused oxidation of some added PPs such as quercetin and myricetin. Size-Exclusion HPLC analysis showed that adding a combination of PP extract and HM pectin decreased the S–H group concentration and increased the percentage of the Unextractable High Molecular Weight (UHMW) proteins, compared to the control bread. The added pectin and PPs, and possibly the oxidation products of the added PPs, affected the interactions between water and bread components (such as gluten proteins) during dough development and bread baking, causing differences in bread cross-linking microstructure and textural properties, which ultimately influenced the extractability and/or stability of added PPs.

Red-fleshed fruit occur in a few taxa of the genus Actinidia. In such taxa, anthocyanins can accumulate in different parts of the fruit including the skin, the whole pericarp or only the inner pericarp. Differences in the relative amounts of cyanidin- and delphinidin-based anthocyanins account for the different shades of red observed. Red-fleshed fruit of A. chinensis accumulate mainly cyanidin 3-Oxylo-galactoside and cyanidin 3-O-galactoside, which usually are restricted to the inner pericarp of the fruit, creating a characteristic and colourful pattern in mature fruit. In order to understand the process of anthocyanin synthesis in Actinidia fruit, we have studied the expression of the anthocyanin biosynthetic genes identified in our Actinidia EST database. Of the different genes of the pathway, only the expression of one glycosyltransferase (F3GT1), confirmed functionally to be cyanidin 3-O-galactosyltransferase, was strongly correlated with red pigmentation of the fruit. However, inheritance of the gene could not be associated with the segregation of the red phenotype of the fruit, suggesting that other factors are able to regulate the biosynthetic pathway. We show that the F3GT1 gene is regulated by a MYB-bHLH mechanism; allelic variation on these genes is likely to account for the expression of the different phenotype.

Much of the diversity of anthocyanins is due to the action of glycosyltransferases, which add sugar moieties to anthocyanidins. We identified two glycosyltransferases, F3GT1 and F3GGT1, from red-fleshed kiwifruit (Actinidia chinensis) that perform sequential glycosylation steps. Red-fleshed genotypes of kiwifruit accumulate anthocyanins mainly in the form of cyanidin 3-O-xylo-galactoside. Genes in the anthocyanin and flavonoid biosynthetic pathway were identified and shown to be expressed in fruit tissue. However, only the expression of the glycosyltransferase F3GT1 was correlated with anthocyanin accumulation in red tissues. Recombinant enzyme assays in vitro and in vivo RNA interference (RNAi) demonstrated the role of F3GT1 in the production of cyanidin 3-O-galactoside. F3GGT1 was shown to further glycosylate the sugar moiety of the anthocyanins. This second glycosylation can affect the solubility and stability of the pigments and modify their colour. We show that recombinant F3GGT1 can catalyse the addition of UDP-xylose to cyanidin 3-galactoside. While F3GGT1 is responsible for the end-product of the pathway, F3GT1 is likely to be the key enzyme regulating the accumulation of anthocyanin in red-fleshed kiwifruit varieties.

Oleic acid (OA) has been used as a control fatty acid in dietary polyunsaturated fatty acid (PUFA) intervention studies due to its lack of effect on eiconasoid biosynthesis. Since the effect of OA as a control fatty acid has not yet been investigated for transcriptomics and proteomics studies, this study aimed to test whether colonic transcriptome and proteome profiles associated with colitis development in mice fed a linoleic acid-rich corn oil-AIN-76A diet (Il10(-/-) compared to C57 mice) where similar to those of OA-fed Il10(-/-) compared to C57 mice (genotype comparison). A close clustering of colonic gene and protein expression profiles between the mice fed the AIN-76A or OA diet was observed. Inflammation-induced regulatory processes associated with cellular and humoral immune responses, cellular stress response and metabolic processes related to energy utilization were identified in Il10(-/-) compared to C57 mice fed either diet. Thus OA was considered as a suitable control unsaturated fatty acid for use in multi-omics PUFA studies. The second aim of this study was to test the effect of an OA-enriched AIN-76A diet compared to a linoleic acid-rich corn oil-AIN-76A diet on colonic transcriptome and proteome changes within Il10(-/-) or C57 mice (diet comparison). Overall, there was a limited concordance observed between measureable transcriptomics and proteomics profiles for genotype and diet comparisons. This underlines the importance and validity of a systems biology approach to understand the effects of diet on gene expression as a function of the genotype.

Epidemiological studies reveal that fruit consumption reduces the prevalence of airway inflammation and childhood asthma. In particular, blackcurrant polyphenolic extracts have been shown to alleviate lung inflammation. Since IL-4-stimulated eotaxin-3 (CCL26) secretion is a major factor in the continuous eosinophil recruitment observed in atopic asthma, our focus was to evaluate the effectiveness of blackcurrant polyphenolic compounds on CCL26 secretion in human alveolar epithelial cells. Our results indicate that a proanthocyanin-enriched blackcurrant extract (BC-P), but not anthocyanin-enriched blackcurrant extract suppressed both IL-4- and IL-13-stimulated CCL26 secretion in a dose-dependent manner. Furthermore pre-incubation of cells with BC-P caused a time-dependent suppression of IL-4-stimulated CCL26 secretion. Moreover, epigallocatechin (EGC), and to a lesser extent epicatechin, metabolites identified in the proanthocyanidin extract, suppressed IL-4-stimulated CCL26 secretion. EGC was also effective at reducing the cellular phosphorylated STAT-6/STAT-6 ratio. Furthermore, both BC-P and purified EGC potentiated the ability of IFN-gamma to suppress IL-4-stimulated CCL26 secretion. The progression of an allergic immune response is complex, identifying plant compounds that target specific cellular events and complement the body's own immune actions is important for the development of functional foods. Our findings support the potential for blackcurrant polyphenolic compounds to reduce eosinophil recruitment and alleviate eosinophilic-driven airway inflammation.

Preeclampsia (PE) is a common, potentially life-threatening pregnancy syndrome triggered by placental factors released into the maternal circulation, resulting in maternal vascular dysfunction along with activated inflammation and coagulation. Currently there is no screening test for PE. We sought to identify differentially expressed plasma proteins in women who subsequently develop PE that may perform as predictive biomarkers. In seven DIGE experiments, we compared the plasma proteome at 20 wk gestation in women who later developed PE with an appropriate birth weight for gestational age baby (n=27) or a small for gestational age baby (n=12) to healthy controls with uncomplicated pregnancies (n=57). Of the 49 differentially expressed spots associated with PE-appropriate for gestational age, PE-small for gestational age or both (p0.05, false discovery rate corrected), 39 were identified by LC-MS/MS. Two protein clusters that accurately (90%) classified women at risk of developing PE were identified. Immunoblots confirmed the overexpression of fibrinogen gamma chain and alpha-1-antichymotrypsin in plasma prior to PE. The proteins identified are involved in lipid metabolism, coagulation, complement regulation, extracellular matrix remodeling, protease inhibitor activity and acute-phase responses, indicating novel synergism between pathways involved in the pathogenesis of PE. Our findings are remarkably similar to recently identified proteins complexed to high-density lipoprotein and linked to cardiovascular disease.

The colour of mangosteen (Garcinia mangostana L.) fruit changes from green to purple black after harvest as the fruit ripens, and is used as a quality guide for growers and consumers. We determined the relationship between anthocyanin composition and content during fruit colour development in relation to fruit maturity and postharvest quality. Fruit at different stages of maturity (light greenish yellow with 5% scattered pink spots to purple black) were harvested and kept at 25 °C (85–90% RH). Fruit from each maturity stage all developed to the final purple black stage. During the postharvest period, hue angle values and pericarp firmness decreased significantly, while soluble solids contents increased. Anthocyanin contents in the outer pericarp were higher than in the inner pericarp and increased to a maximum at the final colour stage. Sensory evaluation and fruit quality (hue angle values, soluble solids and titratable acidity) of fruit harvested at the different stages did not differ once the fruit had finally developed to the purple black stage. The anthocyanins in the outer pericarp mainly consisted of five compounds, identified by HPLC/MS as cyanidin-sophoroside, cyanidin-glucoside, cyanidin-glucoside-pentoside, cyanidin-glucoside-X, cyanidin-X2 and cyanidin-X, where X denotes an unidentified residue of m/z 190, a mass which does not correspond to any common sugar residue. Cyanidin-3-sophoroside and cyanidin-3-glucoside were the major compounds and the only ones that increased with fruit colour development.