Increased understanding of the cereal phytase complement for better mineral bio-availability and resource management

The present paper summarizes the current state of knowledge on cereal phytase that are particular relevant for improving mineral and phosphate bio-availability. Phytases can initiate the hydrolysis of phytate, the main storage form of phosphate in cereals and the major anti-nutritional factor for the bio-availability of micronutrients in human nutrition. The composition and levels of mature grain phytase activity (MGPA) in cereals is of central importance for efficient phytate hydrolysis. The MGPA varies considerably between species. Substantial activity is present in Triticeae tribe cereals like wheat, barley and rye whereas non-Triticeae cereals such as maize and rice have very little MGPA. Recent studies have determined the evolutionary relationships of phytases in Triticeae and non-Triticeae and highlighted the importance of the purple acid phosphatase phytases (PAPhys). In the Triticeae, PAPhys are synthesized during grain development (PAPhy_a) and during germination (PAPhy_b). In non-Triticeae species, only PAPhys that are mainly synthesized during germination were identified. The new knowledge provides new opportunities for modulating the MGPA in Triticeae cereals but also indicates that nutritionally relevant levels of MGPA are unlikely to be achieved in non-Triticeae by conventional breeding. Increasing MGPA in barley via cisgenesis is discussed.     

Manufacture and characterization of pasta made with wheat flour rendered gluten-free using fungal proteases and selected sourdough lactic acid bacteria

Wheat flour, which was rendered gluten-free by sourdough lactic acid bacteria fermentation and fungal proteases, was used for manufacturing experimental gluten-free pasta (E-GFp), according to a traditional process with low temperature drying cycle. Chemical, technological, structural, nutritional and sensory features were characterized and compared with those of commercial gluten-free (C-GFp) and durum wheat pasta (C-DWp). As shown through immunological analyses, the residual concentration of gluten of the hydrolyzed wheat flour was below 10 ppm. E-GFp showed rapid water uptake and shorter optimal cooking time compared to the other pastas. Despite the absence of the gluten network, the supplementation with pre-gelatinized rice flour allowed structural properties of E-GFp, which were comparable to those of C-GFp. The in vitro protein digestibility of E-GFp resulted the highest. Probably due to proteolysis during sourdough fermentation; chemical scores, essential amino acid profile, biological value and nutritional index of E-GFp were higher than those of C-DWp. The hydrolysis index (HI) of E-GFp was ca. 30% lower than that found for C-GFp. As shown by sensory analysis, the characteristic of E-GFp were acceptable. The manufacture of E-GFp should be promising to expand the choice of gluten-free foods, which combine sensory and nutritional properties.     

Alternative matrices for cortisol measurement in fish

Plasma cortisol is the most commonly used indicator of stress in fish but, as the blood sampling procedure itself can be a source of stress, it would be helpful to measure cortisol using less invasive matrices. It is also necessary to find alternative matrices as stress indicators in dead fish in which blood sampling is impossible. In the present study, we investigated transport stress in three aquaculture species, European sea bass (Dicentrarchus labrax L.), common carp (Cyprinus carpio L.) and rainbow trout (Oncorhynchus mykiss Walbaum), by cortisol determination (radioimmunoassay) in plasma and other matrices (skin mucus, gut content, lateral muscle and caudal fin). Cortisol significantly increased after transport in all species and matrices, except in the sea bass gut content, where it remained unchanged. The three species responded to transport stress by producing different cortisol levels. In conclusion, the significant correlation found between plasma cortisol and most of the other matrices opens up the possibility of using them to evaluate stress in fish: mucus sampling is a less invasive method than blood sampling, and in addition to muscle and fin sampling, it can be used in postmortem fish.     

Using Static and Dynamic Indicators to Evaluate Soil Physical Quality in a Sicilian Area

Both capacitive indicators derived from the water retention curve and dynamic measurements of the flow‐weighted mean pore radius, R₀, were used to assess the soil physical quality of two agricultural areas (cropland and olive orchard) and two natural areas (grassland and managed woodlot plantation) potentially subject to soil degradation. The overall idea of the study was to investigate whether a dynamic indicator quantitatively derived from hydraulic conductivity measurements could be used to supplement the traditionally applied capacitive indicators retrieved from water retention measurements. According to the available criteria, only the surface layer of the cropland site showed optimal soil physical quality. In the grassland and woodlot sites, the physical quality was deteriorated also as a consequence of compaction because of grazing. Overall, the physical quality was better in tilled than nontilled soils. The optimal soil in terms of capacitive indicators had hydraulic conductivity close to saturation that was intermediate among the different land uses, and it remained 1·3–1·9 times higher than that observed in the natural sites even when the largest pores emptied. A depth effect on R₀ was observed only when larger macropores were activated. It was suggested that water transmission parameters are more affected by changes in large pore domain. The plant available water content and Dexter's S‐index showed inverse statistically significant regressions with R₀. The empirical relationships were physically convincing given that, at increasing R₀, the contribution of macropores increases, water is transmitted faster below the root zone and the soil's ability to store water is reduced. Copyright © 2013 John Wiley & Sons, Ltd.     

Identification of anthocyanins in Rhamnus alaternus L. berries

Anthocyanin composition in berries of Rhamnus alaternus L., a perennial wild shrub typical of the Mediterranean area, was determined for the first time. The pigments were extracted from the berries with 0.1% HCl in methanol and purified using a C-18 solid-phase cartridge. High-performance liquid chromatography diode array detection-mass spectrometry analysis showed that delphinidin 3-O-rutinoside represented about 62.4% of the total pigments. Other anthocyanins were 3-O-rutinoside derivatives of cyanidin (8.4%), petunidin (15.8%), pelargonidin (4.7%), and peonidin and malvidin (8.7%). The concomitant presence of the six most common anthocyanidins suggested that R. alaternus berries, besides being a good pigment source, could also be a useful tool for anthocyanin identification.     

The soil skeleton, a forgotten pool of carbon and nitrogen in soil

To evaluate the contribution of rock fragments to the soil's total carbon content, the soil of 26 sites, ranging from the Canadian Arctic to the Jordan desert, was analysed for the content of organic C and total N in both fine earth and skeleton fractions. The soils, uncultivated and cultivated, are derived from 11 parent materials: sandstone, mica-schist, granite, gneiss, basaltic pyroclastites, trachyte, dolomite, beach deposits, clay schist, marl and serpentinite. For each soil horizon the contents of fine earth and skeleton were determined by volume. Both fractions were analysed for bulk density, total and organic C and total N. Our results indicate that rock fragments contain amounts of C and N that depend on the nature of the parent material and on its resistance to the weathering processes. The C and N of both fine earth and skeleton were used to calculate the contents of these elements for three depths. At each depth, the skeleton contributes C and N to the soil depending on its abundance. We conclude that the contribution of the rock fragments to the soil C and N cannot be predicted from the soil taxa, but can from the parent material. Calculations that exclude C and N of the skeleton could lead to errors in the estimates of these two elements in soils.     

Development of new fungicides against Magnaporthe grisea: synthesis and biological activity of pyrazolo[3,4-d][1,3]thiazine,pyrazolo[1,5-c][1,3,5]thiadiazine,and pyrazolo[3,4-d]pyrimidine derivatives

Some pyrazolo[3,4-d]pyrimidin-4(5H)-thione, pyrazolo[3,4-d][1,3]thiazin-4-one/thione, and pyrazolo[1,5-c][1,3,5]thiadiazine-4-one/thione derivatives were synthesized and screened for antifungal activity against the causal agent of rice blast disease, Magnaporthe grisea. In all cases a remarkable inhibition of fungal growth was found in the range from 10 to 200 microg x mL(-1). Several compounds were able to control mycelium growth at a rate of 10 microg x mL(-1), a concentration at which the reference compound tricyclazole was completely ineffective. At least in the case of the most active substance, at the same dose the growth of seedlings or cultured cells of rice was substantially unaffected. Results allowed definition of structural requirements either to maintain or to enhance mycotoxic activity.     

Changes in phenolic composition and antioxidant activity of virgin olive oil during frying

The concentration of hydroxytyrosol (3,4-DHPEA) and its secoiridoid derivatives (3,4-DHPEA-EDA and 3,4-DHPEA-EA) in virgin olive oil decreased rapidly when the oil was repeatedly used for preparing french fries in deep-fat frying operations. At the end of the first frying process (10 min at 180 degrees C), the concentration of the dihydroxyphenol components was reduced to 50-60% of the original value, and after six frying operations only about 10% of the initial components remained. However, tyrosol (p-HPEA) and its derivatives (p-HPEA-EDA and p-HPEA-EA) in the oil were much more stable during 12 frying operations. The reduction in their original concentration was much smaller than that for hydroxytyrosol and its derivatives and showed a roughly linear relationship with the number of frying operations. The antioxidant activity of the phenolic extract measured using the DPPH test rapidly diminished during the first six frying processes, from a total antioxidant activity higher than 740 micromol of Trolox/kg down to less than 250 micromol/kg. On the other hand, the concentration of polar compounds, oxidized triacylglycerol monomers (oxTGs), dimeric TGs, and polymerized TGs rapidly increased from the sixth frying operation onward, when the antioxidant activity of the phenolic extract was very low, and as a consequence the oil was much more susceptible to oxidation. The loss of antioxidant activity in the phenolic fraction due to deep-fat frying was confirmed by the storage oil and oil-in-water emulsions containing added extracts from olive oil used for 12 frying operations.     

A neutralizing antibody selected from plasma cells that binds to group 1 and group 2 influenza A hemagglutinins

The isolation of broadly neutralizing antibodies against influenza A viruses has been a long-sought goal for therapeutic approaches and vaccine design. Using a single-cell culture method for screening large numbers of human plasma cells, we isolated a neutralizing monoclonal antibody that recognized the hemagglutinin (HA) glycoprotein of all 16 subtypes and neutralized both group 1 and group 2 influenza A viruses. Passive transfer of this antibody conferred protection to mice and ferrets. Complexes with HAs from the group 1 H1 and the group 2 H3 subtypes analyzed by x-ray crystallography showed that the antibody bound to a conserved epitope in the F subdomain. This antibody may be used for passive protection and to inform vaccine design because of its broad specificity and neutralization potency.