A comparative study between responses of isolated bovine and equine digital arteries to vasoactive mediators

Hemodynamic perturbations, partly resulting from abnormal vasoconstriction of digital vessels, have been implicated in the pathogenesis of bovine and equine laminitis. This study compared the responsiveness of isolated bovine (BDA) and equine (EDA) digital arteries to pharmacological agents that stimulate receptor systems involved in the regulation of normal vessel tone. The role of the endothelium and the short‐ and longer‐term effects of an experimentally induced endothelial damage were also evaluated. Species‐related differences were found in the vessel reactivity to all of the receptor agonists tested. In intact BDA, as compared to intact EDA, norepinephrine was a more effective vasoconstrictor, 5‐hydroxytryptamine a more effective but less potent vasoconstrictor, isoproterenol a less effective vasodilator and carbamylcholine a less potent vasodilator. In BDA, but not in EDA, the contractile responses to norepinephrine and 5‐hydroxytryptamine were enhanced immediately after endothelium removal. However, the contractile reactivity of denuded BDA returned to basal values following overnight incubation. The differences suggest species specificity for the pathophysiology of digital vasomotor tone and function in horses and cattle.     

Haematological, biochemical and immunological parameters as stress indicators in Dicentrarchus labrax and Sparus aurata farmed in off-shore cages

In 2000–2001, an investigation was performed in two Sicilian mariculture sites (Pachino and Castellammare) to monitor physiological and biochemical parameters in sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax) specimens farmed in off-shore sea cages. Plasma cortisol and glucose levels, tissue lactate concentration, haematocrit and haemoglobin content, lysozyme, haemolytic and haemagglutinating activities were determined. During the experiment, an increase in tissue lactate and plasma cortisol levels and a reduction in haemolytic and haemagglutinating titers, were recorded. Changes occurring in these haematological, immunological and biochemical values suggest that these parameters can be useful indicators in assessing the condition of chronic stress induced by mariculture operations.     

Metallomics approach to trace element analysis in ustilago maydis using cellular fractionation, atomic absorption spectrometry, and size exclusion chromatography with ICP-MS detection

Huitlacoche is the ethnic name of the young fruiting bodies of Ustilago maydis, a common parasite of maize. In Mexico and other Latin American countries, this fungus has been traditionally appreciated as a local delicacy. In this work a metallomics approach was used with the determination of eight elements in huitlacoche by electrothermal atomic absorption spectrometry as one facet of this approach. The results obtained indicated relatively lower concentrations of commonly analyzed metals, as referred to the data reported for other mushroom types. This effect was ascribed to different accessibilities of elements, depending on fungus substrate (lower from plant than from soil). Subcellular fractionation was accomplished by centrifugation of cell homogenates suspended in Tris-HCl buffer. Recoveries of the fractionation procedure were in the range of 71-103%. For six elements (Cr, Cu, Fe, Mn, Ni, and Pb), the mean relative contributions in cytosol, cell walls, and mixed membrane fraction were 50.7, 48.2, and 1.1% respectively. To attain the molecular weight distribution of compounds containing target elements as an additional aspect of the metallomics approach, the fungus extract (1% sodium dodecyl sulfate in Tris-HCl, 30 mmol L(-)(1), pH 7.0) was analyzed by size exclusion chromatography with UV and ICP-MS detection. With spectrophotometric detection (280 nm), the elution of high molecular weight compounds was observed in the form of one peak (MW > 10 kDa), and several lower peaks appeared at higher retention times (MW < 10 kDa). On ICP-MS chromatograms, a coelution of (59)Co, (63)Cu, (57)Fe, (202)Hg, (60)Ni, and (80)Se with the first peak on the UV chromatogram was clearly observed, indicating that a fraction of each element incorporated with high molecular weight compounds (12.7, 19.8, 33.7, 100, 19.4, and 45.8%, respectively, based on the peak area measurements). From a comparison of (80)Se and (33)S chromatograms (for sulfur analysis, the extract was obtained in the absence of SDS), both elements coeluted with the first UV peak, but their lower molecular weight compounds were apparently different. These findings may contribute to a better understanding of the accumulation of elements in mushrooms.     

Development of a methodology to forecast the nutritional value of new tomato hybrids

Tomato is a food of great relevance both due to its large-scale consumption and its richness in bioactive components. It is an important component of the traditional Mediterranean diet as well as of other diets. Nowadays, concerns about human health and environment preservation have changed the objectives of tomato breeding. In this study, eight tomato F₁ hybrids and their parental lines were analyzed for nutritional properties and agronomic traits using a new selection method that combines biochemical and agronomic evaluation. Eight traits contributing to the nutritional quality of tomato (lycopene, β-carotene, other carotenoids, flavonoids, phenolic acids, vitamins C and E, dry residue) and average yield were assessed in fifteen tomato genotypes. Furthermore the pathogen resistances possessed from these genotypes were, also, considered. In order to select valuable tomato hybrids, a nutritional index (I_QUAN) and an agronomic index (AI) were calculated. Our results suggested that the I_QUAN nutritional index may be very useful to forecast the nutritional value of F₁ hybrids based on parental performance. Combining the use of the I_QUAN and AI indexes, we were able to select two hybrids (MR 48 and MR 47) that contain considerable amounts of antioxidants and acceptable parameters for commercial production.     

Effects of endotoxin and influence of cyclooxygenase-2 on β-adrenergic mediated relaxation in isolated equine digital artery

The effects of endotoxin on β-adrenergic-mediated relaxation were investigated in the equine digital artery (EDA). Possible involvement of cyclooxygenase-2 (COX-2) in endotoxin-induced effects and basal EDA β-adrenoceptor functionality was also evaluated. Endothelium-intact (e(+)) and/or -denuded (e(-)) EDA rings were incubated overnight with lipopolysaccharide (LPS), LPS+NS398 (selective COX-2 inhibitor) or NS398 alone. Vessel rings were then mounted in organ baths and relaxant responses to isoproterenol (ISOP) recorded on U44069-induced pre-contraction. Response to ISOP was further evaluated in either incubated or freshly isolated (e(-)) rings acutely exposed to NS398. Fresh and incubated (e(-)) EDAs were also analysed for COX-2 expression by Western blotting. LPS caused endothelium-dependent enhancement of β-adrenergic mediated relaxation. NS398 did not reverse endotoxin effects, suggesting that COX-2 did not have a mediating role. In the absence of LPS, NS398 significantly increased ISOP-induced relaxation. This finding, together with immunoblot detection of COX-2 in both fresh and incubated (e(-)) vessels, revealed the existence of a constitutive COX-2 exerting tonic inhibitory modulation on EDA β-adrenergic-mediated relaxation. The results support the possible role of endotoxin in the vascular disturbances associated with equine laminitis. Moreover, the involvement of COX-2 in the physiological regulation of EDA tone warrants further clinical investigation into the efficacy and safety of selective COX-2 inhibitors on digital circulation in horses.     

Characterization of selenium species in Brazil nuts by HPLC-ICP-MS and ES-MS

Brazil nuts have been classified as the foodstuffs that contain the highest level of unadulterated selenium, an essential trace element that appears to prevent cancer. To date, characterization of the selenium species in brazil nuts has not yet been investigated. In this work, various sample preparation approaches, including microwave extractions and enzymatic treatments, are examined with the goal of species preservation and subsequent selenium speciation; of these approaches, an enzymatic treatment with Proteinase K proved most effective. High-performance liquid chromatography (HPLC) separation strategies and inductively coupled plasma mass spectrometry (ICP-MS) detection schemes will also be presented. Extracts are evaluated against available standards for the commercially obtainable seleno-amino acids, selenomethionine (SeMet), selenoethionine (SeEt), and selenocystine (SeCys); selenomethionine was demonstrated to be the most abundant of these seleno-amino acids. Further characterization of unidentified selenium-containing peaks is attempted by the employment of several procedures, including electrospray-mass spectrometry (ES-MS). A peptide structure was identified; however, this was considered a tentative proposal due to the large background produced by the extremely complicated brazil nut matrix.     

Water Quality Simulation for Planning Restoration of a Mined Watershed

Water quality simulation was performed to evaluate the effects of restoration alternatives on metals transport in a mountainous watershed in Montana, U.S.A. impacted by hundreds of abandoned hardrock mines. The Water Quality Analysis Simulation Program (WASP5), developed by the U.S. Environmental Protection Agency (U.S. EPA), was used to assist in planning restoration of the Upper Tenmile Creek watershed, a major drinking water supply for the City of Helena. Synoptic survey data collected by U.S. EPA and the U.S. Geological Survey were used for model calibration and validation. The effectiveness of eight restoration alternatives was modeled under steady-state, low flow conditions. These alternatives ranged from removal of adit and point source discharges to modification of the water supply scheme to provide higher in-stream flows. The model was also used for a number of related purposes, including evaluation of metals loadings and losses, exceedances of water quality standards, interactions between metals in water and bed sediment, and model and data uncertainties. Although standards exceedances are common throughout the watershed, modeling results indicated that removal of point sources, mine waste near watercourses, and streambed sediment can help improve water quality. Alteration of the water supply scheme and increasing baseflow will also ultimately be required to meet standards for all metals. The model also showed that although adits and point sources contribute significant metals loadings to the stream during baseflow, in some areas shallow groundwater and bed sediment can also be sources of metals. Adsorption and precipitation onto bed sediments are also important loss mechanisms in some locations. The model helped to identify uncertainties in the metal partition coefficients associated with sediment, significance of precipitation reactions, and locations of unidentified sources and losses of metals.