Dissecting the impact of environment, season and genotype on blackcurrant fruit quality traits.

This work aims to determine the effect of genotype x environment (GxE) interaction that influence blackcurrant (Ribes nigrum) fruit quality. We applied metabolomics-driven analysis on fruits from four cultivars grown in contrasting European-locations over two seasons. By integrating metabolomics and sensory analysis, we also defined specific metabolic signatures associated with consumer acceptance. Our results showed that rainfall is a crucial factor associated with accumulation of delphinidin- and cyanidin-3-O-glucoside, the two mayor blackcurrant pigments meanwhile temperature affects the main organic acid levels which can be decisive for fruit taste. Sensorial analysis showed that increases in terpenoid and acetate ester volatiles were strongly associated with higher appreciation score, while proacacipetalin, a cyanogenic-glycoside, was positively associated to bitter taste. Our results pave the way for the selection of high-quality cultivars and suitable production sites for blackcurrant cultivation.

Qualitative phytochemical analysis and in vitro investigation of the immunomodulatory properties of honeys produced in Kazakhstan.

Honey is known to have antimicrobial, immunomodulatory and wound healing properties. The biological properties of honey have been attributed to phytochemicals derived from their source plants and research has focused on identifying the bioactive phytochemicals with therapeutic potential. In this study, we determined the ability of 5 honeys from Kazakhstan and manuka honey to stimulate TNF-α and TGF-ß production by human keratinocytes. TNF-α and TGF-ß levels increased over time in honey treated and untreated keratinocytes, whereas cells treated with sugar solutions that matched those of the honeys had reduced levels of both cytokines. This suggests that the non-sugar phytochemical components of the honeys may have prevented this decrease. Analysis by LC-MS confirmed that the honeys contained a diverse range of phytochemicals. Some phytochemicals e.g. pinobanksin and vanillin were present at different levels across the honey types, whereas other components, e.g. dicarboxylic acids and their glycosides, were abundant in all honeys.

Polysaccharides from European Black Elderberry Extract Enhance Dendritic Cell Mediated T Cell Immune Responses.

European black elderberry (Sambucus nigra L.) is a popular way to treat common colds or influenza infections. Mechanistically, this might be due to a direct antiviral effect or a stimulatory effect on the immune system of the host. Here, we evaluated the modulatory effects of black elderberry derived water extract (EC15) and its polysaccharide enriched fractions (CPS, BOUND, and UNBOUND) in comparison to a conventional alcoholic extract (EE25), regarding the phenotypical and functional properties of dendritic cells (DCs), which are essential cells to induce potent T cell responses. Interestingly, the water extract and its polysaccharide fractions potently induced DC maturation, while the ethanol extract did not. Moreover, the capacity to stimulate T cells by these matured DCs, as assessed using MLR assays, was statistically higher when induced by the water extracted fractions, compared to immature DCs. On the other hand, the ethanol extract EE25 did not induce T cell stimulation. Finally, the cytokine expression profiles of these DC-T cell cocultures were assessed and correlated well with increased T cell stimulation. Also, the expression of inflammatory cytokines, such as IL-6, TNF-α, and IFN-γ was highly increased in the presence of the elderberry water extract EC15, and the polysaccharide enriched CPS, BOUND, and UNBOUND fractions, but not by EE25. Thus, from these data, we conclude that the polysaccharides present in water-derived elderberry fractions induce potent immune-modulatory effects, which represents the basis for a strong immune-mediated response to viruses including influenza.

Identification of components in Kazakhstan honeys that correlate with antimicrobial activity against wound and skin infecting microorganisms.

BACKGROUND: Antimicrobial drug resistance is a major public health threat that can render infections including wound and skin infections untreatable. The discovery of new antimicrobials is critical. Approaches to discover novel antimicrobial therapies have included investigating the antimicrobial activity of natural sources such as honey. In this study, the anti-microbial activity and chemical composition of 12 honeys from Kazakhstan and medical grade manuka honey were investigated. METHODS: Agar well diffusion and broth culture assays were used to determine anti-microbial activity against a range of skin and wound infecting micro-organisms. Folin-Ciocalteu method was used to determine the total phenol content of the honeys and non-targeted liquid chromatography analysis was performed to identify components that correlated with antimicrobial activity. RESULTS: In the well diffusion assay, the most susceptible micro-organisms were a clinical isolate of Methicillin resistant Staphylococcus aureus (MRSA) and Enterococcus faecalis (ATCC 19433). Buckwheat & multi-floral honey from Kazakhstan demonstrated the highest antimicrobial activity against these two micro-organisms. Kazakhstan honeys with a buckwheat floral source, and manuka honey had the highest total phenol content. Non-targeted liquid chromatography analysis identified components that correlated with anti-microbial activity as hydroxyphenyl acetic acid, p-coumaric acid, (1H)-quinolinone, and abscisic acid. CONCLUSIONS: The Kazakhstan honeys selected in this study demonstrated antimicrobial activity against wound and skin infecting micro-organisms. Compounds identified as correlating with antimicrobial activity could be considered as potential bioactive agents for the treatment of wound and skin infections.

Extraction, Enrichment, and LC-MSn-Based Characterization of Phlorotannins and Related Phenolics from the Brown Seaweed, Ascophyllum nodosum.

Phenolic components from the edible brown seaweed, Ascophyllum nodosum, have been associated with considerable antioxidant activity but also bioactivities related to human health. This study aims to select and identify the main phlorotannin components from this seaweed which have been previously associated with potential health benefits. Methods to enrich phenolic components then further select phlorotannin components from ethanolic extracts of Ascophyllum nodosum were applied. The composition and phenolic diversity of these extracts were defined using data dependent liquid chromatography mass spectroscopic (LC-MSn) techniques. A series of phlorotannin oligomers with apparent degree of polymerization (DP) from 10 to 31 were enriched by solid phase extraction and could be selected by fractionation on Sephadex LH-20. Evidence was also obtained for the presence of dibenzodioxin linked phlorotannins as well as sulphated phlorotannins and phenolic acids. As well as diversity in molecular size, there was evidence for potential isomers at each DP. MS2 fragmentation analyses strongly suggested that the phlorotannins contained ether linked phloroglucinol units and were most likely fucophlorethols and MS3 data suggested that the isomers may result from branching within the chain. Therefore, application of these LC-MSn techniques provided further information on the structural diversity of the phlorotannins from Ascophyllum, which could be correlated against their reported bioactivities and could be further applied to phlorotannins from different seaweed species.

Glutathionyl-S-chlorogenic acid is present in fruit of Vaccinium species, potato tubers and apple juice.

A hydroxycinnamate-like component was identified in highbush blueberry (Vaccinium corymbosum) fruit, which had identical UV and mass spectrometric properties to an S-linked glutathionyl conjugate of chlorogenic acid synthesized using a peroxidase-catalyzed reaction. The conjugate was present in fruits from all highbush blueberry genotypes grown in one season, reaching 7-20% of the relative abundance of 5-caffeoylquininc acid. It was enriched, along with anthocyanins, by fractionation on solid phase cation-exchange units. Mining of pre-existing LC-MS data confirmed that this conjugate was ubiquitous in highbush blueberries, but also present in other Vaccinium species. Similar data mining identified this conjugate in potato tubers with enrichment in peel tissues. In addition, the conjugate was also present in commercial apple juice and was stable to pasteurization and storage. Although glutathionyl conjugates of hydroxycinnamic acids have been noted previously, this is the first report of glutathionyl conjugates of chlorogenic acids in commonly-eaten fruits and vegetables.

Cinnamon Shows Antidiabetic Properties that Are Species-Specific: Effects on Enzyme Activity Inhibition and Starch Digestion.

Despite considerable research the evidence around the antidiabetic properties of cinnamon remains equivocal, and this may be due to varietal differences which is an aspect that is understudied. This study systematically compared the anti-hyperglycaemic properties of the four major commercial cinnamon types used around the world (Chinese; Cinnamomum cassia [CC], Indonesian; C. burmanii [IC], Vietnamese; C. loureirii [VC], and Ceylon; C. zeylanicum [SC]). LC-MS analysis showed distinct diffrences in the phytochemical profiles of cinnamon with SC showing the lowest coumarin concentration. CC and IC had the highest polyphenol levels and antioxidant potential, and all four types differed significantly in their content (P < 0.001). All cinnamon types showed potent species-specific effects on starch digestion enzyme activity inhibition (P < 0.001), CC was most effective against α-amylase and all four strongly inhibited α-glucosidase compared to acarbose. Cinnamon significantly reduced starch breakdown during oral (P = 0.006) and gastric (P = 0.029) phases of gastro-intestinal digestion with IC and SC showing consistent effects. No effects of cinnamon were seen in the intestinal phase. IC, VC and SC showed the greatest potential to inhibit formation of advanced glycation endproducts (AGEs) during digestion. In conclusion, cinnamon demonstrates anti-hyperglycaemic properties, however effects are species-specific with best overall properties seen for Ceylon cinnamon.

Potato miR828 Is Associated With Purple Tuber Skin and Flesh Color.

Anthocyanins are plant pigments responsible for the colors of many flowers, fruits and storage organs and have roles in abiotic and biotic stress resistance. Anthocyanins and polyphenols are bioactive compounds in plants including potato (Solanum tuberosum L.) which is the most important non-cereal crop in the world, cultivated for its tubers rich in starch and nutrients. The genetic regulation of the flavonoid biosynthetic pathway is relatively well known leading to the formation of anthocyanins. However, our knowledge of post-transcriptional regulation of anthocyanin biosynthesis is limited. There is increasing evidence that micro RNAs (miRNAs) and other small RNAs can regulate the expression level of key factors in anthocyanin production. In this study we have found strong associations between the high levels of miR828, TAS4 D4(-) and purple/red color of tuber skin and flesh. This was confirmed not only in different cultivars but in pigmented and non-pigmented sectors of the same tuber. Phytochemical analyses verified the levels of anthocyanins and polyphenols in different tissues. We showed that miR828 is able to direct cleavage of the RNA originating from Trans-acting siRNA gene 4 (TAS4) and initiate the production of phased small interfering RNAs (siRNAs) whose production depends on RNA-dependent RNA polymerase 6 (RDR6). MYB transcription factors were predicted as potential targets of miR828 and TAS4 D4(-) and their expression was characterized. MYB12 and R2R3-MYB genes showed decreased expression levels in purple skin and flesh in contrast with high levels of small RNAs in the same tissues. Moreover, we confirmed that R2R3-MYB and MYB-36284 are direct targets of the small RNAs. Overall, this study sheds light on the small RNA directed anthocyanin regulation in potato, which is an important member of the Solanaceae family.

Extracts from the edible seaweed, Ascophyllum nodosum, inhibit lipase activity in vitro: contributions of phenolic and polysaccharide components.

A polyphenol-rich extract (PRE) from the edible seaweed, Ascophyllum nodosum, inhibited pancreatic lipase activity in an oil-based turbidimetric assay with an IC50 of 200 µg gallic acid equivalents (GAE) perassay) [∼230 µg DW] whereas the known inhibitor, Orlistat, gave an IC50 at 0.4 µg per assay. A phlorotannin-enriched fraction (TRF) purified from the PRE was more potent with an IC50 = 60 µg GAE per assay (∼65 µg DW). When the assay was started by the addition of lipase, both Orlistat and TRF were much less effective which suggests that pre-incubation of enzyme and inhibitor improved inhibition. Based on phenol content, water extracts from Ascophyllum were more potent lipase inhibitors than PRE (IC50 ∼ 150 µg GAE per assay). However, this was equivalent to ∼580 µg DW and these extracts contained polysaccharides (e.g. alginate content = 110 µg mL-1) which may also contribute to inhibition. Indeed, a polysaccharide-enriched fraction obtained by ethanol precipitation gave an IC50 of 1000 µg DW which was equivalent to 130 µg GAE and 420 µg alginate per assay. Therefore a >3 fold increase in alginate content did not markedly improve inhibition. Re-precipitation increased alginate content and reduced polyphenol content but lipase inhibition was markedly reduced (i.e. IC50 at ∼1100 µg DW per assay, 700 µg alginate and 25 µg GAE). Purifying the polysaccharide fraction by ion exchange removed all phenolics but the IC50 increased to >2500 µg DW, equivalent to >1970 µg alginate per assay. In conclusion, polysaccharides and phlorotannins may inhibit lipase in an additive fashion, with phlorotannins apparently more effective in vitro. However, interactions between these components may be important when food products containing this edible seaweed are consumed.