Inhibition of inducible nitric oxide synthase expression by an acetonic extract from Feijoa sellowiana Berg. fruits

Feijoa sellowiana Berg. fruits and especially the acetonic extract have been shown to possess biological activities, although the responsible compounds have never been identified. The present study was designed to evaluate the anti-inflammatory activity of an acetonic extract from F. sellowiana Berg. fruits on the nitric oxide (NO) pathway, which plays an important role in inflammation. To this aim the J774 cell line, which expresses inducible nitric oxide synthase (iNOS) following stimulation with lipopolysaccharide (LPS), has been utilized, and the effects of this extract and its fractions on NO production, iNOS protein expression, and signal pathways involved in its regulation have been evaluated. This study demonstrates that at least some part of the anti-inflammatory activity of the acetonic extract is due to the suppression of NO production by flavone and stearic acid. The mechanism of this inhibition seems to be related to an action on the expression of the enzyme iNOS through the attenuation of nuclear factor kappaB (NF-kappaB) and/or mitogen-activated protein kinase (MAPK) activation.     

Lipoxygenase distribution in coffee (Coffea arabica L.) berries

In this paper lipoxygenase (LOX) presence was investigated in coffee berries to determine its involvement in lipid degradative metabolism of plants grown in organic and conventional cultivations. An immunochemical analysis has evidenced a ca. 80 kDa protein, cross-reacting with an anti-LOX antibody, only in the pulp fraction of berries obtained from plants of both cultivations. LOX activity in this fraction could be monitored either as conjugated diene formation or reaction products (determined by HPLC) and was mainly associated with a heavy membrane fraction (HMF, enriched in tonoplast, endoplasmic reticulum, plasma membrane, and mitochondria) and a light membrane fraction (LMF, enriched in plasma membrane and endoplasmic reticulum, with low levels of tonoplast and mitochondria). The LOX activity of LMF from berries of both cultivations showed an optimum at pH 8.0. The HMF exhibited a different activity peak in samples from conventional (pH 8.0) and organic (pH 5.5) cultures, suggesting the presence of different isoenzymes. These findings were also confirmed by variation of the ratio of 9- and 13-hydroperoxides in organic (1:1) and conventional cultivations (1:10), indicating that the organic one was subjected to an oxidative stress in the coffee pulp fraction leading to the expression of an acidic LOX. Such de novo synthesized LOX activity could be responsible for the production of secondary metabolites, which may interfere with the organoleptic profile of coffee.     

You cannot conserve a species that has not been found: The case of the marine sponge Axinella polypoides in Liguria,Italy

1. Detailed knowledge about the distribution of species in need of protection is required for the management of Marine Protected Areas, a major tool to reduce marine biodiversity loss. Such knowledge is deficient for most marine invertebrates.
2. Axinella polypoides is a marine sponge included on the list of protected species by the Barcelona Convention (1976) and the Bern Convention (1987). This large and erect species has an important ecological role in habitat forming and benthic–pelagic coupling.
3. Bathymetrical, geographical and ecological data over the last 60 years were collated from publications and reports, together with new surveys to assess the distribution and protection status in Liguria of A. polypoides. It identified a more widespread distribution than previously thought, which points at a general need for dedicated investigations on the occurrence of species that require protection.
4. Bathymetrical distribution was trimodal, with peaks corresponding to different geomorphological settings: coastal cliff bases (around 38 m depth), inner shelf shoals (52 m) and rocks amidst coarse sediment on the outer shelf (79 m). Density was significantly greater at the shallowest depths. The species was mostly found in the coralligenous biocoenosis, in association with other characteristic species or forming a monospecific facies.
5. On (sub)vertical cliffs, A. polypoides often exhibited an unusual cane shape, rather than the typical bushy morphology, thus causing confusion with the congeneric A. cannabina, a more southern species. Records of the latter in the Ligurian Sea therefore need confirmation.
6. Only a minority (22.6%) of A. polypoides records were in Marine Protected Areas, the remainder being located in areas with no current environmental protection plans in place. While the occurrence of this species in MPAs remained stable over the decades, the only quantitative historical data available indicated that populations in non-protected areas were declining owing to anthropogenic impacts (fishing and anchoring).

     

Reconstructing the history of the sand tiger shark (Carcharias taurus) in the Mediterranean Sea

1. Sharks are globally exposed to several anthropogenic threats, which, in many cases, have severely reduced their distribution and have impacted populations. In the Mediterranean Sea, because of its long history of exploitation and the relatively short span of scientific monitoring, reconstructing shark baselines is challenging. Many vulnerable species declined in population abundance and geographic distribution before it was possible to adequately track these changes. Consequently, for many of these species, current conservation assessments are now suffering from a severe case of shifting baseline syndrome, whereby their historical occurrence in the area is questioned.
2. The sand tiger shark (Carcharias taurus) is one of these cases. Characterized by high philopatry, restricted home range, and low interchange between populations, its Mediterranean population may have been severely impacted by the high historical fishing pressure in the region before scientific monitoring began.
3. In this study, the history of the abundance and distribution of C. taurus in the Mediterranean Sea was reconstructed through a comprehensive search of occurrence records in the literature and in museum collections. Between 1810 and 2008, 31 occurrence records and 54 publications provided information on the presence of the species in the area. These records were sparse but systematic over time and indicated occurrence hot spots in the south‐western Mediterranean Sea and in the eastern Adriatic Sea.
4. The presence of ten sightings of juveniles from a total of 18 sightings with length information suggested local parturition. Habitat and extinction models indicate that the area has suitable oceanographic conditions for the occurrence of the species and that the species cannot be considered extinct in the Mediterranean Sea. Our study suggests that there is still hope for the recovery of the species and underlines the crucial role of historical investigations to reconstruct the history of large elasmobranchs in the Mediterranean Sea.

     

Space–time patterns of co‐variation of biodiversity and primary production in phytoplankton guilds of coastal marine environments

• 1. The relevance of biodiversity to ecosystem processes is a major topic in ecology. Here, we analyse the relationship between biodiversity and productivity of the nano‐ and micro‐phytoplankton guilds in coastal marine ecosystems.
• 2. The patterns of variation of species richness, diversity and primary productivity (as ¹⁴C assimilation) were studied in two marine areas: a eutrophic–mesotrophic area beside the River Po delta (northern Adriatic) and an oligotrophic area around the Salento peninsula (southern Adriatic–Ionian). The study was carried out at 23 sites in the northern area and at 45 sites in the southern area. Sites were arranged on expected spatial and temporal gradients of primary productivity variation, according to distance from the coast, optical depths and seasonal period.
• 3. 167 taxa were identified in the northern area and 153 taxa in the southern area. In both areas, the taxonomic composition of the nano‐ and micro‐phytoplankton guilds exhibited greater temporal than spatial variation. The latter was much higher in the southern area than in the northern area (average dissimilarity between stations being 70.7±0.8% and 44.7±4.2% respectively).
• 4. Primary productivity varied in space and time on the gradients considered. Phytoplankton species richness and diversity exhibited significant patterns of variation in space and time; overall, these were inversely related to the primary productivity patterns in the northern area, whereas they were directly related in the southern area.
• 5. The small individual size and the high turnover rate of phytoplankton are likely to underlie the observed relationships, which emphasized a threshold response to nutrient enrichment in agreement with the ‘paradox of enrichment’. Under resource enrichment conditions, the high turnover of producers leads to hierarchical partitioning of the available resources with an increasing dominance of a few species. Therefore, the relationship observed here seems likely to be explained by the complementarity hypothesis.

Copyright © 2003 John Wiley & Sons, Ltd.     

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.     

Hydrolysis of cellobiose by β-glucosidase in the presence of soil minerals - Interactions at solid-liquid interfaces and effects on enzyme activity levels

Extracellular enzymatic activities in soils are essential for the cycling of organic matter. These activities take place in multiphase environments where solid phases profoundly affect biocatalytic activities. Aspergillus niger is ubiquitous in soils; its β-glucosidase plays an important role in the degradation of cellulose, and therefore in the global carbon cycle and in the turnover of soil organic matter. However, the information on the interactions of this protein with soil minerals is very limited, and even less is known about their consequences for the hydrolysis of the natural substrate cellobiose. We therefore characterised the sorptive interactions of this enzyme with the soil minerals montmorillonite, kaolinite and goethite and quantified the resulting changes in the hydrolysis rate of cellobiose. Fractions of adsorbed protein, and the resulting catalytic activity loss, were lower for montmorillonite than for kaolinite and goethite at given experimental conditions; adsorption was 9.7 ± 7.3% for montmorillonite, 70.3 ± 3.1% for kaolinite and 71.4 ± 1.8% for goethite, respectively. Adsorption of the protein to the minerals caused a total decrease in the catalytic activity of 18.8 ± 3.4% for kaolinite and 17.9 ± 4.7% for goethite whereas it was not significant for montmorillonite. The average catalytic activity lost by the pool of adsorbed molecules was 26.8% for kaolinite and 25.0% for goethite. Both the amount of adsorbed protein and the resulting loss of catalytic activity were found to be independent of the specific surface areas yet were influenced by the electrical properties of the mineral surfaces. Under the experimental conditions, montmorillonite and kaolinite are negatively charged whereas goethite is positively charged. However, because of the adsorption of phosphate anions from the buffer, a charge reversal took place at the surface of goethite. This was confirmed by zeta (ζ)-potential measurements in phosphate buffer, revealing negative values for all the tested minerals. Indeed goethite interacted with the enzyme as a negatively charged surface: the amount of adsorbed protein and the resulting catalytic activity loss were very similar to those of kaolinite. Our results show that, even if an important fraction of β-glucosidase is adsorbed to the minerals, the catalytic activity is largely retained. We suggest that this strong activity retention in presence of soil minerals results from a selective pressure on A. niger, which benefits from the activity of the adsorbed, and thus stabilized, enzyme pool.     

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.     

Comparative analysis of a wood: adhesive bondline

The wood–adhesive interface was analyzed using five methods with the objective of quantitatively assessing penetration of adhesive into the porous wood network. Methods included fluorescence microscopy, scanning electron microscopy, backscatter electron imaging, wavelength dispersive spectroscopy, and X-ray microtomography (XMT). Each method provided a visual inspection, and all of the analysis methods were applied to the same field of view. XMT was the primary technique of interest. Rubidium hydroxide was used in place of sodium hydroxide in the formulation of phenol–formaldehyde adhesive. Rubidium was found to increase the X-ray attenuation of the adhesive. However, rubidium migrated beyond the adhesive interphase during specimen preparation, thus reducing its effectiveness for image contrast enhancement. The wood species studied included red oak (Quercus rubra), Douglas-fir (Pseudotsuga menziesii), and hybrid poplar (Populus deltoides × Populus trichocarpa). All techniques used for this study were useful, but each presented some limitations for bondline analysis. Despite the problem with rubidium migration, XMT for this application was promising.     

A hierarchical model for the analysis of spatial rainfall extremes

In this article, we propose a spatial model for analyzing extreme rainfall values over the Triveneto region (Italy). We assess the existence of a long-term trend in the extremes. To integrate data coming from the different stations, we propose a hierarchical model. At the first level, for each monitoring station we model data by making use of a generalized extreme value distribution; at the second level, we combine results from the first stage by exploiting recent advances in modeling nonstationary spatial random fields.