The stability of three strawberry cultivars was evaluated for changes in jam color quality during processing and storage at 20°, 30° and 37°C for 200 days. Anthocyanin content was determined by HPLC. The effect of cultivar, processing and storage on jam pigments, instrumental color (L*, a*, b*) and consumer preference were also determined. ‘Oso grande’ jam had the lowest anthocyanin concentration (110 mg/g f.w), higher monomeric pigment degradation during processing and storage, highest pH, least desirable color score from the sensory panel and shortest shelf‐life. Similarities were found between jams prepared with ‘Chandler'and Tudla’ cultivars, as well as initial differences in total anthocyanin concentrations (195 and 130 mg/g f.w.).

Ion trap mass spectrometry (ITMS) was used to obtain further qualitative information about the chemical composition of humic-like substances (HULIS) in atmospheric particulate matter. Particles ≤10 µm (PM 10) were collected on quartz fiber filters for 24 h in the region of Basel (Switzerland) and extracted with water. HULIS were separated from inorganic salts by size exclusion chromatography (SEC) and detected by electrospray ionization in the negative ion mode (ESI(−)). Series of consecutive fragment ion spectra (MS n) were recorded by ITMS. Full scan mass spectra of the extracts showed a mass distribution pattern characteristic for HULIS. Different molecular ions were selected from this pattern for further fragmentations. Among them the molecular ion m/z 299 was considered as representative and intensively studied. Many MS 2 and MS 3 fragment spectra contained a fragment m/z 97 and a neutral loss of 80 u. Time-of-flight (TOF) MS and deuterium exchange experiments identified m/z 97 as hydrogen sulfate. MS 2 and MS 3 fragment spectra supported the existence of sulfate covalently bound to HULIS. The fragmentation behavior of sulfated HULIS could be confirmed by model compounds.

Knowledge of the molecular basis of plant resistance to pathogens in species other than Arabidopsis is limited. The function of Fa WRKY1, the first WRKY gene isolated from strawberry (Fragaria x ananassa), an important agronomical fruit crop, has been investigated here. Fa WRKY1 encodes a IIc WRKY transcription factor and is up-regulated in strawberry following Colletotrichum acutatum infection, treatments with elicitors, and wounding. Its Arabidopsis sequence homologue, At WRKY75, has been described as playing a role in regulating phosphate starvation responses. However, using T-DNA insertion mutants, a role for the At WRKY75 and Fa WRKY1 in the activation of basal and R-mediated resistance in Arabidopsis is demonstrated. At wrky75 mutants are more susceptible to virulent and avirulent isolates of Pseudomonas syringae. Overexpression of Fa WRKY1 in At wrky75 mutant and wild type reverts the enhanced susceptible phenotype of the mutant, and even increases resistance to avirulent strains of P. syringae. The resistance phenotype is uncoupled to PATHOGENESIS-RELATED (PR) gene expression, but it is associated with a strong oxidative burst and glutathione-S-transferase (GST) induction. Taken together, these results indicate that At WRKY75 and Fa WRKY1 act as positive regulators of defence during compatible and incompatible interactions in Arabidopsis and, very likely, Fa WRKY1 is an important element mediating defence responses to C. acutatum in strawberry. Moreover, these results provide evidence that Arabidopsis can be a useful model for functional studies in Rosacea species like strawberry.

Important losses in strawberry production are caused by species of the fungus Colletotrichum, the causal agent of anthracnose. However, very limited studies at molecular level exist of the mechanisms related to strawberry susceptibility against this pathogen. We have analysed a moderately resistant cultivar (cv. Andana) together with a very susceptible one (cv. Camarosa) during the process of infection with Colletotrichum acutatum at a molecular level. To gain insight into this interaction we have identified a large number of strawberry genes involved in signalling, transcriptional control, defence and many genes with unknown function with altered expression in response to C. acutatum infection. Spatial and temporal gene expression profiles after infection showed that the response was dependant on the tissue and cultivar analysed and also quicker and/or stronger in the moderately resistant cultivar (cv. Andana) than in the susceptible one (cv. Camarosa). Interestingly, we found that genes described as being induced during pathogen infection such as g-thionins, peroxidases, chitinases and b-1-3-glucanases were downregulated in fruit and/ or crown tissues of the very susceptible cultivar. Our results yielded a first insight on some of the genes responding to this plant-pathogen interaction at molecular level and suggest that pathogen progression can be dependent upon a reduction of the active defences of strawberry and this is genotype and tissue dependent.

Dendritic cells (DCs) constitute the first point of contact between gut commensals and our immune system. Despite growing evidence of the immunomodulatory effects of probiotics, the interactions between the cells of the intestinal immune system and bacteria remain largely unknown. Indeed,, the aim of this work was to determine whether the probiotic Bifidobacterium breve CNCM I-4035 and its cell-free culture supernatant (CFS) have immunomodulatory effects in human intestinal-like dendritic cells (DCs) and how they respond to the pathogenic bacterium Salmonella enterica serovar Typhi, and also to elucidate the molecular mechanisms involved in these interactions. Human DCs were directly challenged with B. breve/CFS, S. typhi or a combination of these stimuli for 4 h. The expression pattern of genes involved in Toll-like receptor (TLR) signaling pathway and cytokine secretion was analyzed. CFS decreased pro-inflammatory cytokines and chemokines in human intestinal DCs challenged with S. typhi. In contrast, the B. breve CNCM I-4035 probiotic strain was a potent inducer of the pro-inflammatory cytokines and chemokines tested, i.e., TNF-α, IL-8 and RANTES, as well as anti-inflammatory cytokines including IL-10. CFS restored TGF-β levels in the presence of Salmonella. Live B.breve and its supernatant enhanced innate immune responses by the activation of TLR signaling pathway. These treatments upregulated TLR9 gene transcription. In addition, CFS was a more potent inducer of TLR9 expression than the probiotic bacteria in the presence of S. typhi. Expression levels of CASP8 and IRAK4 were also increased by CFS, and both treatments induced TOLLIP gene expression. Our results indicate that the probiotic strain B. breve CNCM I-4035 affects the intestinal immune response, whereas its supernatant exerts anti-inflammatory effects mediated by DCs. This supernatant may protect immune system from highly infectious agents such as Salmonella typhi and can down-regulate pro-inflammatory pathways.

Probiotic bacteria have been shown to modulate immune responses and could have therapeutic effects in allergic and inflammatory disorders. However, little is known about the signalling pathways that are engaged by probiotics. Dendritic cells (DCs) are antigen-presenting cells that are involved in immunity and tolerance. Monocyte-derived dendritic cells (MoDCs) and murine DCs are different from human gut DCs; therefore, in this study, we used human DCs generated from CD34+ progenitor cells (hematopoietic stem cells) harvested from umbilical cord blood; those DCs exhibited surface antigens of dendritic Langerhans cells, similar to the lamina propria DCs in the gut. We report that both a novel probiotic strain isolated from faeces of exclusively breast-fed newborn infants, Lactobacillus paracasei CNCM I-4034, and its cell-free culture supernatant (CFS) decreased pro-inflammatory cytokines and chemokines in human intestinal DCs challenged with Salmonella. Interestingly, the supernatant was as effective as the bacteria in reducing pro-inflammatory cytokine expression. In contrast, the bacterium was a potent inducer of TGF-β2 secretion, whereas the supernatant increased the secretion of TGF-β1 in response to Salmonella. We also showed that both the bacteria and its supernatant enhanced innate immunity through the activation of Toll-like receptor (TLR) signalling. These treatments strongly induced the transcription of the TLR9 gene. In addition, upregulation of the CASP8 and TOLLIP genes was observed. This work demonstrates that L. paracasei CNCM I-4034 enhanced innate immune responses, as evidenced by the activation of TLR signalling and the downregulation of a broad array of pro-inflammatory cytokines. The use of supernatants like the one described in this paper could be an effective and safe alternative to using live bacteria in functional foods.

No abstract

The fragmentation behaviour of seven pairs of isomeric flavone/isoflavone aglycones (solely hydroxylated and/or methoxylated) was studied using ion trap mass spectrometry with atmospheric pressure ionisation (API, both electrospray and APCI) in the positive and negative ion modes. A major difference was found in the neutral loss of 56 u, which was a common feature of all isoflavones in API(+). It was identified as a double loss of CO by accurate mass tandem mass spectrometric (MS/MS) measurements using a hybrid quadrupole time-of-flight (Q-TOF) instrument. Fragmentation of daidzein with (13)C-isotope labelling of the carbon C2 showed that this double loss occurred from the central ring of the molecule. A mechanism for this selective fragmentation is given. Further isoflavone-specific fragmentations were used to develop a guideline for the identification of isoflavone structures. A software-based neutral loss scan of 56 u in the API(+)-MS(2) mode was applied to extracts of leaves of Lupinus albus and to soy flour. The structure elucidation guideline allowed identification of hydroxy and/or methoxy isoflavones. Structures could be confirmed for those available as reference compounds.