Evaluation of phenolic compounds in virgin olive oil by direct injection in high-performance liquid chromatography with fluorometric detection

Hydrophilic phenols are the most abundant natural antioxidants of virgin olive oil (VOO), in which tocopherols and carotenes are also present. The prevalent classes of hydrophilic phenols found in VOO are phenyl alcohols, phenolic acids, secoiridoids such as the dialdehydic form of decarboxymethyl elenolic acid linked to (3,4-dihydroxyphenyl)ethanol or (p-hydroxypheny1)ethanol (3,4-DHPEA-EDA or p-HPEA-EDA) and an isomer of the oleuropein aglycon (3,4-DHPEA-EA), lignans such as (+)-1-acetoxypinoresinol and (+)-pinoresinol, and flavonoids. A new method for the analysis of VOO hydrophilic phenols by direct injection in high-performance liquid chromatography (HPLC) with the use of a fluorescence detector (FLD) has been proposed and compared with the traditional liquid-liquid extraction technique followed by the HPLC analysis utilizing a diode array detector (DAD) and a FLD. Results show that the most important classes of phenolic compounds occurring in VOO can be evaluated using HPLC direct injection. The efficiency of the new method, as compared to the liquid-liquid extraction, was higher to quantify phenyl alcohols, lignans, and 3,4-DHPEA-EA and lower for the evaluation of 3,4-DHPEA-EDA and p-HPEA-EDA.     

Assessment of geographic variation by RAPD markers among Italian open-pollinated progenies of Populus alba L.

Random amplified polymorphic DNA (RAPD) analysis wasapplied to evaluate variation among provenances of white poplar(Populus alba L.) collected indifferent geographic areas in Italy. Fifteen decamer oligonucleotidesamplified a total of 71 distinct fragments of which 57(80% of the total observed) were polymorphic among53 plants of white poplar. Among provenances, the highest degree ofpolymorphism was detected in the population from Sinni river(62%), while the lowest level was recorded in thepopulation from Arno river (42%). Cluster(UPGMA) analysis was performed on the RAPD data and itdifferentiated the analysed plants in three distinct groupscorresponding to the Apennine, Southern and Northern Italian areas.The RAPD clusters were congruent with groups based on phenologicaltraits.     

Intergeneric relationships in the Australian Vitaceae: new evidence from cpDNA analysis

Taxa related to important agricultural species are likely to contain a considerable amount of potentially valuable genetic diversity. Nevertheless, before breeding programs or gene discovery projects can be initiated it is important to understand the phylogenetic relationships between the species involved. A component of a major gene discovery project in grapes at the Centre for Plant Conservation Genetics (Southern Cross University, Australia) is directed at the discovery of novel genes in native Vitaceae. As a result a study was conducted in order to assess the phylogenetic relationships between V. vinifera and the native members of the three major Australian genera: Cayratia, Cissus and Tetrastigma. CpDNA sequence analysis (from the trnL intron) adequately resolved intergeneric relationship between the majority of the species studied and provided some useful new information on the phylogenetic relationships within the Vitaceae. This preliminary project identified two species, C. hypoglauca and C. sterculiifolia, as being closely related to V. vinifera and worthy of further in-depth investigation.     

On the chemical composition and nutritional value of pleurotus taxa growing on umbelliferous plants (apiaceae)

Unpublished data on the chemical composition and nutritional value of Pleurotus mushrooms, growing on Umbelliferous plants (Apiaceae), are here reported. Cultivated basidiomata of four different Pleurotus taxa were analyzed in order to evaluate the composition in lipids, sugars, nitrogen, water, vitamins, ashes, and energetic values. The results showed that Pleurotus mushrooms are suitable in every type of diet thanks to their low caloric content, gastronomic value, vitamins, and mineral salt contents. The presence of a high content of vitamin B(12) and riboflavin in Pleurotus nebrodensis is noteworthy.     

The impact of clonality on an endangered tree (Elaeocarpus williamsianus) in a fragmented rainforest

The occurrence and distribution of clonality in the endangered rainforest tree Elaeocarpus williamsianus (Elaeocarpaceae) was investigated using SSR and RAPD analyses for 170 apparent individual trees found across seven sites. The results obtained with the two molecular techniques were in complete agreement in showing that single clones are present in most of the E. williamsianus sites with two genets occurring at the largest and most adequately protected site. In addition, seed production, viability and germinability were determined for four of these populations. Fruit were produced in all four populations tested although sterile fruit were very common. Only two E. williamsianus trees representing different genets within the same site produced viable seed. The overall genetic diversity within E. williamsianus is much lower than expected and thus the potential for sexual reproduction has been significantly diminished. It is concluded that habitat fragmentation removed an existing balance between vegetative and sexual reproduction in this species. Such findings have added urgency to the management of this species which could include a reintroduction program incorporating the collation of all clones within selected sites.     

Effects of the aglycone of ascaulitoxin on amino acid metabolism in Lemna paucicostata

Ascaulitoxin and its aglycone (2,4,7-triamino-5-hydroxyoctanoic acid, CAS 212268-55-8) are potent phytotoxins produced by Ascochyta caulina, a plant pathogen being developed for biocontrol of weeds. The mode of action of this non-protein amino acid was studied on Lemna paucicostata. Ascaulitoxin is a potent growth inhibitor, with an I₅₀ for growth of less than 1 μM, almost completely inhibiting growth at about 3 μM. Its action is slow, starting with growth inhibition, followed by darker green fronds, and then chlorosis and death. Most amino acids, including non-toxic non-protein amino acids, reversed the effect of the toxin when supplemented in the same medium. Supplemental sucrose slightly increased the activity. d-Amino acids were equally good inhibitors of ascaulitoxin activity, indicating the amino acid effects may not be due to inhibition of amino acid synthesis. Oxaloacetate, the immediate precursor of aspartate, also reversed the activity. LC-MS did not detect interaction of the compound with lysine, an amino acid that strongly reversed the effect of the phytotoxin. Metabolite profiling revealed that the toxin caused distinct changes in amino acids. Reduction in alanine, paralleled by enhanced levels of the branched chain amino acids valine, leucine and isoleucine and nearly unchanged levels of pyruvate, might indicate that the conversion of pyruvate to alanine is affected by ascaulitoxin aglycone. In addition, reduced levels of glutamate/glutamine and aspartate/asparagine might suggest that synthesis and interconversion reactions of these amino group donors are affected. However, neither alanine aminotransferase nor alanine: glyoxylate aminotransferase were inhibited by the toxin in vitro. Our observations might be explained by three hypotheses: (1) the toxin inhibits one or more aminotransferases not examined, (2) ascaulitoxin aglycone affects amino acid transporters, (3) ascaulitoxin aglycone is a protoxin that is converted in vivo to an aminotransferase inhibitor.     

Assessment of mast cells degranulation in rainbow trout (Oncorhynchus mykiss Walbaum) by means of gray level and texture analysis (Gray Level Correlation Matrices)

Degranulation of intestinal mast cells in rainbow trout (Oncorhynchus mykiss Walbaum) was studied by means of image analysis technique. Two strips from the same intestinal segment from ten clinically healthy trout were sampled. One was immediately mounted in an organ bath, treated with compound 48/80 and then processed, as the remnant untreated strip, for light microscopy. Colour pictures taken from each section were converted in their gray levels RGB stacks equivalent and in 8 bits gray levels. Five granular cytoplasm areas of mast cells (MCs) for each section were analysed for the following parameters: mean gray values and Gray Level Correlation Matrices parameters. All RGB channels and the 8 bit gray levels transformed images showed higher mean gray level values after compound 48/80 exposure (Anova, p<0.01). Only two channels (Red and Green) and the 8 bit gray levels transformed images of a unique texture parameter (contrast) appeared to be significantly higher after compound 48/80 exposure (Anova, p<0.05). Red channel and Blue channel means gray level values were able to discriminate between not degranulated and degranulated MCs. The described model represents a promising alternative in mast cell research.     

The effects of restriction of movement on the reliability of heart rate variability measurements in the horse (Equus caballus)

Analysis of heart rate variability (HRV) is a noninvasive approach for investigating the sympathovagal balance of the autonomic nervous system. In recent years, HRV has been increasingly evaluated in animal research. In horses, it has been suggested that basal resting conditions can be achieved by restraining them. The aim of this study was to verify how restriction of movement influences HRV i2n horses. Ten healthy standardbred mares were used to measure the electrocardiographic signal under 2 conditions: free to move in the stall and restrained in the stock. Results indicate that the restriction of movement is associated with increased nervous system sympathetic activity not consistent with resting conditions.     

Applications of high-resolution solid-state NMR spectroscopy in food science

The principal applications of high-resolution solid-state NMR spectroscopy, in the field of food science, are reviewed, after a short general introduction, mainly focusing on the potential of these investigations, which are, today, routine tools for resolving technological problems. Selected examples of the applications in the field of food science of high-resolution solid-state NMR spectroscopy both in (13)C and in (1)H NMR particularly illustrative of the results obtainable are reported in some detail.