Studies on the Glutathione-Dehydroascorbate Oxidoreductase (EC 1.8.5.1) from Wheat Flour

Glutathione (GSH) dehydrogenase was partially purified from wheat flour after extraction, ammonium sulfate precipitation, and ionic-exchange chromatography on diethylaminoethyl (DEAE) Sepharose CL6B. Kinetic studies showed that the optimum pH was close to 7.5. The K_m values varied between 0.15 and 0.28 mM for dehydroascorbic acid (DHA) and between 1.8 and 0.62 mM for GSH when pH was varied from 5.5 to 7.5. The kinetic pattern was consistent with a sequential mechanism for the binding of GSH and DHA. NaCl is a competitive inhibitor with respect to GSH and is uncompetitive with respect to DHA, which suggests that the enzyme combines with DHA before it does with GSH. IsoDHA can replace DHA as hydrogen acceptor but with a K_m of 1.2 mM. γ-Glu-cys was enzymically oxidized but much less efficiently than GSH (V_m = 47 nkat/mL and K_m = 5.5 mM compared to V_m = 362 nkat/mL and K_m = 1.8 mM for GSH), whereas cysteine and cys-gly were not substrates. In the presence of DHA, addition of cysteine and cys-gly to small amounts of GSH causes a large activation of the enzymatic formation of ascorbic acid suggesting coupled oxidation of these thiols.     

Where land meets sea: Intertidal areas as key-habitats for sharks and rays

Intertidal habitats (i.e. marine habitats that are (partially) exposed during low tide) have traditionally been studied from a shorebird-centred perspective. We show that these habitats are accessible and important to marine predators such as elasmobranchs (i.e. sharks and rays). Our synthesis shows that at least 43 shark and 45 ray species, of which 54.5% are currently threatened, use intertidal habitats. Elasmobranchs use intertidal habitats mostly for feeding and as refugia, but also for parturition and thermoregulation. However, the motivation of intertidal habitat use remains unclear due to limitations to observe elasmobranch behaviour in these dynamic habitats. We argue that elasmobranch predators can play an important role in intertidal food webs by feeding on shared resources during high tide (i.e. ‘high-tide predators’), which are accessible and also consumed by terrestrial or avian predators during low tide (i.e. ‘low-tide predators’). In addition, elasmobranchs are able to change the bio-geomorphology of intertidal habitats by increasing habitat heterogeneity due to feeding activities and may also alter resource availability for other consumers. We discuss how the ecological role of elasmobranchs in intertidal habitats is being affected by the continued overexploitation of these species, and conversely, how the global loss of intertidal areas poses an additional threat to an already vulnerable taxonomic group. We conclude that studies on intertidal ecology should include both low-tide (e.g. shorebirds) and high-tide (e.g. elasmobranchs) predatory guilds and their ecological interactions. The global loss of elasmobranch predatory species and intertidal habitat provides additional compelling arguments for the conservation of these areas.     

Antioxidant Capacity Analysis of Blackberry Extracts with Different Phytochemical Compositions and Optimization of their Ultrasound Assisted Extraction

High antioxidant capacity molecules, such as anthocyanins, occur naturally in blackberry (Rubus fruticosus). In particular, ‘Dasha’ blackberry cultivar has scarcely been studied and, it is becoming economically more important in Mexico. In this study, several blackberry extraction conditions, conferred different amounts of detectable phytochemical groups that in turn were analysed with a new approach to investigate their influence on antioxidant capacity (AC). Additionally, a central composite design (CCD) was proposed to study effects of temperature and acidification on AC. Finally, an original approach was used to disclose interactions between the phytochemical content and the AC. Changes in size of the particles during extraction were reported for the first time, and the results showed evidence of swelling and dissolving of particles. UAE of fine and thick powders achieved similar efficiencies in contrast with maceration which showed large differences for the extraction of the tested sizes. CCD showed that low levels of acidification and high levels of temperature resulted in higher extraction of phytochemicals and AC. HPLC show that the main anthocyanidin may represent 88% of the total anthocyanins. Due to its relatively high abundance, cyanidyn-3-glucoside showed evidence of being the main cause of the changes in AC in ‘Dasha’ extracts. The use of mild conditions resulted in no degradation of anthocyanins and, therefore, there are no AC losses. A correlation plane was proposed to study synergisms of the extracts with other anthocyanins or phenolics.     

Factors influencing half‐pearl (mabe) production and quality in the red abalone Haliotis rufescens

The factors influencing half‐pearl (mabe) production in the red abalone Haliotis rufescens were investigated, as a strategy to optimize the technology for the development of a pearling industry in Chile. The effects of abalone size (small, large), implant position (dorsal, central, ventral and their combinations), and number of nuclei (one, two, three) were analysed on the quantity, thickness of the nacre layer and quality (shape, colour, lustre, defects) of the mabe produced. The size of abalone did not exert a clear influence on the quantity and quality of mabe, but the position on the shell (firstly) and number of nuclei (secondly) did. More mabe, with thicker nacre, brighter lustre and fewer defects occurred in small or large abalone implanted with only one nucleus on dorsal position. In contrast, small and large abalone implanted in the ventral position formed less mabe, with thinner nacre, poor lustre and more defects. This likely occurs because mantle tissue folds at the ventral position and does not always cover the nucleus, leaving it partially or totally exposed. Most of the pearls were categories AA in small abalone and A in large abalone; only three AAA gems were produced in large abalone with one nucleus on dorsal position or with three nuclei (DCV). Based on these results, we recommend implanting abalone at 60–100 mm shell length, with only one nucleus on the dorsal position.     

Flavonoids and biflavonoids in Tuscan berries of Juniperus communis L.: detection and quantitation by HPLC/DAD/ESI/MS

The aim of the present work was to develop a quali-quantitative investigation, using HPLC/DAD and HPLC/ESI/MS techniques, of the phenolic composition of berries collected from wild Tuscan plants of Juniperus communis L. and grown in three different geographical zones. The applied chromatographic elution method made it possible to well separate up to 16 different compounds belonging to flavonoids, such as isoscutellarein and 8-hydroxyluteolin or hypolaetin glycosides, and six biflavonoids, among them amentoflavone, hynokiflavone, cupressoflavone, and methyl-biflavones. To the best of the authors' knowledge this is the first report on the presence of these compounds in juniper berries. The flavonoidic content in the analyzed berries ranged between 1.46 and 3.79 mg/g of fresh pulp, whereas the amount of the biflavonoids was always lower, varying between 0.14 and 1.38 mg/g of fresh weight.     

Phenols content and 2-D electrophoresis protein pattern: a promising tool to monitor <Emphasis Type="Italic">Posidonia</Emphasis> meadows health state

Background  

The endemic seagrass Posidonia oceanica (L.) Delile colonizes soft bottoms producing highly productive meadows that play a crucial role in coastal ecosystems dynamics. Human activities and natural events are responsible for a widespread meadows regression; to date the identification of "diagnostic" tools to monitor conservation status is a critical issue. In this study the feasibility of a novel tool to evaluate ecological impacts on Posidonia meadows has been tested. Quantification of a putative stress indicator, i.e. phenols content, has been coupled to 2-D electrophoretic protein analysis of rhizome samples.     

Antioxidant effect of two virgin olive oils depends on the concentration and composition of minor polar compounds

In vitro studies show that some individual minor polar phenolic compounds (MPC) present in virgin olive oil prevent oxidation of human low-density lipoproteins (LDL), but few data are available on the antioxidant effect of whole oil extract. Thus, whole virgin olive extracts were studied to determine whether they maintain the antioxidant activity and whether this last is linked to MPC composition of a single virgin oil. Using HPLC-DAD the MPC content in Taggiasca and Seggianese virgin olive oils was measured. Taggiasca oil was less rich in total MPC (208.5 mg/L) than Seggianese oil (441.9 mg/L). In addition, the major compounds of Taggiasca oil were lignan derivatives, whereas the major compounds in Seggianese oils were secoiridoid derivatives. Moreover, Taggiasca oil was practically free of 5-hydroxytyrosol and 5-hydroxytyrosol derivatives, deacetoxy-oleuropein aglycone and oleuropein aglycone. The antioxidant activity of the oils on human LDL was evaluated by measuring malondialdehyde and conjugate diene generation induced by copper ions. In both tests, the oil extracts dose-dependently reduced malondialdehyde and conjugate diene generation. Moreover, antioxidant potency correlated with total MPC; thus, Seggianese extract was more active. The two oils differed quantitatively and qualitatively, and these differences influenced their biological activities; thus clinical trials focused on studying the effects of olive oils should specify the oils used.     

Anatomical and biochemical trait network underlying genetic variations in tomato fruit texture

Firmness is an indicator of fruit freshness and a main component of tomato (Solanum lycopersicum) fruit texture. In this work, the genetic variability in fruit firmness and stiffness was analyzed in pre- and postharvest periods and underlying anatomical and biochemical traits were identified. Three tomato contrasted parental lines and six derived quantitative trait loci (QTL)-NILs harboring texture QTL on chromosome 4 (QTL4) and 9 (QTL9) were analyzed; the seasonal variability was assessed on two distant trusses. Firmness and stiffness were measured by compression and puncture tests at harvest and after 7-day storage at 20 °C. QTL4 poorly influenced the textural variables, on the contrary to QTL9 which increased firmness measured by puncture test and had similar effects in the two genetic backgrounds. According to this test, firmness increased along the season, but ranks among genotypes and QTL effects were hardly affected. Only some of the QTL effects were still significant after storage and firmness losses were not predicted by firmness at harvest. Fruit firmness and stiffness measured by puncture tests correlated with both morphological (locular number, R = −0.89), histological (cell size, R < −0.80) and biochemical (dry matter (R > 0.82) and soluble sugar content (R < −0.74)) fruit traits. In contrast, compression test values hardly correlated with any of the measured traits. This work provided an original comprehensive approach to analyse fleshy fruit firmness and paves the way for a future predictive model.     

Staphylococcus aureus proteins differentially recognized by the ovine immune response in mastitis or nasal carriage

Staphylococcus aureus is an opportunistic pathogen in dairy ruminants where it is found in healthy carriage and can be a major cause of mastitis. A better knowledge of the host-pathogen interactions is needed to tackle this serious animal health problem. This study aimed at identifying S. aureus proteins differentially expressed by S. aureus in nasal colonization versus mastitis. Serological proteome analysis (SERPA) was used to examine protein samples prepared from culture supernatants of S. aureus strains originally isolated from gangrenous mastitis and nasal carriage (O11) or subclinical mastitis (O46) and to compare patterns of immune-reactive proteins. These staphylococcal proteins were revealed by sera obtained from ewes suffering from S. aureus mastitis and by sera obtained from healthy nulliparous ewes (i.e. no lactation and no mastitis or other symptoms) that were nasally colonized by S. aureus. Altogether 49 staphylococcal immune-reactive proteins were identified in this study. Patterns of proteins revealed by sera from infected- or healthy carrier- animals were comparable and analysis singled out one immune-reactive protein, N-acetylmuramyl-L-alanine amidase, which was recognized by each of the 6 sera from infected animals, when tested individually, and not by the sera of healthy carriers. This is the first study that compares the S. aureus seroproteome in colonization versus mastitis context in ruminants. These results open avenues for studies aiming at a better understanding of the balance between infection and commensal lifestyle in this opportunistic pathogen and at new prevention strategies.