Isolation and biochemical characterization of a basic myrosinase from ripe Crambe abyssinica seeds,highly specific for epi-progoitrin

On the basis of previous studies on the mechanism-based inhibition, activation, and active site structure of myrosinase(s) isolated from Sinapis alba and other cruciferous seeds, crambe myrosinase shows uncommon properties and behavior. For this reason homogeneous crambe myrosinase was isolated and investigated to establish the most important physicochemical features, including kinetic properties determined with the epimers progoitrin (R) and epi-progoitrin (S) as substrates, with and without ascorbate as an activator. The results of this study demonstrate that crambe myrosinase is highly specific for epi-progoitrin due to a better stabilization of the enzyme-substrate complex. This stabilization is caused by additional hydrogen bonding that only epi-progoitrin can set up between its hydroxyl group and a suitable residue in the hydrophobic pocket where the "docking" of the glucosinolates side chain takes place.     

8S globulin of mungbean [Vigna radiata (L.) Wilczek]: cloning and characterization of its cDNA isoforms, expression in Escherichia coli, purification, and crystallization of the major recombinant 8S isoform

Three isoforms of the cDNA of the major 8S globulin of mungbean, 8Salpha, 8Salpha', and 8Sbeta, were isolated, cloned, and characterized. The cDNA sequences of 8Salpha, 8Salpha', and 8Sbeta had open reading frames of 1362, 1359 or 1362, and 1359 bp, respectively, which code for 454, 453 or 454, and 453 amino acids corresponding to molecular weights of 51 973, 51 627 or 51 758, and 51 779, respectively. Homology in terms of cDNA and amino acid sequences was 91-92% between 8Salpha and 8Salpha', 87% between 8Salpha and 8Sbeta, and 86-88% between 8Salpha' and 8Sbeta. The signal peptide was found to be 1-25, 1-24 or 25, and 1-23 for 8Salpha, 8Salpha', and 8Sbeta, respectively, using the signalP website (Nielsen, H.; Engelbrecht, J.; Brunak, S.; von Heijne, G. Protein Eng. 1997, 10, 1-6). The propeptide was determined to be IVHREN. A single site for glycosylation (N-X-S/T) was observed about 90 amino acids from the C terminus. Homology between mungbean 8S isoforms and other 7-8S proteins ranged from 45 to 68% within members of the legume family and 29 to 34% for crops of different species. The major isoform 8Salpha was expressed in Escherichia coli and purified by successive ammonium sulfate fractionation, hydrophobic interaction, and Mono Q column chromatography. The recombinant 8Salpha, but not the native form, was successfully crystallized producing rhombohedral crystals.     

Mercury contamination in the Brazilian Amazon. Environmental and occupational aspects

Mercury (Hg) contamination of miners, riparian and Indian populations and fish in the Amazon region, due to gold extracting activities, has been studied. Samples of hair, urine, and blood of Indians and prospectors, and hair from riparian fish-eating population and fishes from Madeira river, respectively, were collected and analyzed by Cold Vapor, Atomic Absorption Spectrometry (CV-AAS) techniques. The results obtained showed that the aquatic food chains in the Amazonian ecosystems are contaminated by methylmercury (MeHg), exposing Indians, prospectors and riverines to thr risk of severe health hazard. The highest levels of contamination, based upon hair analysis, were found in riparian of the Madeira river, followed by Cuniã Lake population, Indians (Fresco river) and prospectors, in that order. Blood analysis showed 59% of the samples from Indians and 33% from prospectors with Hg contents above 10 ng mL^?1. Analysis of urine, on the other hand, showed 44% of Indians with Hg levels below the detection limit (d.l.), and 30% above 20 ng mL^?1, whereas 38% of the prospectors presented Hg concentrations over 20 ng mL^?1, and 20% below the d.l. These results prove that prospectors, who hardly ever eat fish but are badly exposed to inorganic Hg vapor, are ocupationally contaminated, while the rest of the populations under survey are exposed to environmental contamination by Hg, through polluted fish ingestion.     

Nitrogen Release from Slow-Release Fertilizers in Soils with Different Microbial Activities

Soil microbial activity is recognized as an important factor affecting nitrogen (N) release from slow-release fertilizers. However,studies on the effect of size and activity of soil microflora on fertilizer degradation have provided contrasting results. To date, no clear relationships exist between soil microbial activity and the release of N from slow-release fertilizers. Hence, the aim of this study was to better understand such relationships by determining the release of N from three slow-release fertilizers in soils with different microbial activities. Soils were amended with urea-formaldehyde (UF), isobutylidene diurea (IBDU), and crotonylidene diurea (CDU). Urea, a soluble fertilizer, was used as the control. Fertilized soil samples were placed in a leaching system, and the release of N was determined by measuring ammonium-N and nitrate-N concentrations in leachates during 90 d of incubation. Non-linear regression was used to fit N leaching rate to a first-order model. In all the treated soils, N was released in the order: urea (89%–100%) IBDU (59%–94%) UF (46%–73%) CDU (44%–56%). At the end of incubation, N released from CDU did not differ (P 0.05) among soils. On the contrary, UF and IBDU released significantly lower (P 0.05) amounts of N in the soil with higher microbial activity and lower pH.The rate constant (K_0) for UF was lower (P 0.05) in the soil with lower pH. Taken together, our results indicated that soil microbial size and microbial activity had a marginal effect on fertilizer mineralization.     

Behavior of Cd,Pb, and Cu in the marine deltaic area of the Po River (North Adriatic Sea)

An analysis of the spatial trend of concentrations and of the correlation between heavy metals and total suspended matter was performed in a multidisciplinary study of transport of pollutants in the marine coastal area involving in situ investigations, remote sensing techniques and modeling. This paper presents results concerning the chemical behavior of heavy metals (Cu, Cd, Pb) in the North Adriatic Sea and in particular in the marine deltaic area of the Po river. The samples were collected during the JRC Remote Sensing Campaigns ADRIA 84 and ADRIA 85, carried out in the entire North Adriatic Sea and in the selected area in front of the Po when passing from the fluvial to the marine environment. In the dissolved phase, Cu undergoes a dilution process consistent with the mixing theory, while there is a tenfold increase in Pb concentrations. A pronounced tendency to augment the level in the dissolved phase is observed for all three elements in the pelagic waters. In the particulate phase, Cd and Cu, after a diminution in the nearshore stations, their concentration increases as the pelagic area is approached. The total amount of heavy metals in solid phase of the water decreases according to the dispersion and the sedimentation processes of the fluvial load in the marine water. A satisfactory correlation was found between the Cu and Pb with the total suspended matter (TSS). An attempt was made to characterize sediments and suspended matter in order to assess the role of the organic and mineralogical components on heavy metal behavior.     

Foot-and-mouth disease vaccination: a multifactorial study of the influence of antigen dose and potentially competitive immunogens on the response of cattle of different ages

Groups of 68 and 66 cattle aged 12 and 24 months respectively were each subdivided into 16 groups and inoculated with foot-and-mouth disease vaccines containing O1 Campos, A24 Cruzeiro and C3 Pando virus strains. The 140S antigen mass of the O1 and A24 valencies was varied while that of C3 was held constant. Multifactorial comparisons between the 21 day serum neutralising antibody titres showed that over most of the range there was a linear log dose response relationship. Doubling the antigen dose increased the serum antibody titres against both A24 Cruzeiro and O1 Campos by approximately 0.15 log10. The A24 antigen was about 30 times more immunogenic than the O1 with C3 intermediate between the two. At high antigen doses the responses flattened but the level at which this occurred depended on the immunogen administered. No difference could be demonstrated between the responses of 12- and 24-month-old cattle and there was no evidence of competitive inhibition or enhancement between the virus strains included in the vaccines.     

Post‐grazing cutting management in tall fescue affects the sward structure,forage and liveweight production

Tall fescue is the main perennial grass of the pastures of the temperate region of Argentina. However, after flowering in spring, tall fescue loses productivity and quality. Based on this, the objective of this work was to evaluate the effects of different post‐grazing mechanical cutting managements on the forage mass, leaf proportion, stocking rate, liveweight gain and liveweight production of tall fescue pastures. The treatments were post‐grazing mechanical cutting at anthesis (FC), post‐grazing mechanical cutting throughout spring and summer (SSC), and no post‐grazing mechanical cutting (NC). The experiment was performed from 2011 to 2014 in Argentina. The greatest and lowest forage mass were determined in September–November and May–September respectively. The leaf proportion of the SSC treatment was greater than that of the NC treatment, except in September–November. NC had higher stocking rate and lower liveweight gain than SSC, and neither NC nor SSC differed from FC. The liveweight production of the treatments was characterized by a trade‐off between stocking rate and liveweight gain. We conclude that FC is an attractive management because, with a single post‐grazing cutting, swards remain productive and leafy.     

Antigenotoxicity and Antioxidant Activity of Acerola Fruit (Malpighia glabra L.) at Two Stages of Ripeness

Genotoxic and antigenotoxic effects of acerola fruit at two stages of ripeness were investigated using mice blood cells. The results show that no ripeness stage of acerola extracts presented any genotoxic potential to damage DNA (Comet assay) or cytotoxicity (MTT assay). When antigenotoxic activity was analyzed, unripe fruit presented higher DNA protection than ripe fruit (red color) extract. The antioxidant capacity of substances also showed that unripe samples inhibit the free radical DPPH more significantly than the ripe ones. The results about determination of compounds made using HPLC showed that unripe acerola presents higher levels of vitamin C as compared to ripe acerola. Thus, vitamin C and the complex mixture of nutrients of Malpighia glabra L., and especially its ripeness stages, influenced the interaction of the fruit extract with the DNA. Acerola is usually consumed when ripe (red fruit), although it is the green fruit (unripe) that has higher potential as beneficial to DNA, protecting it against oxidative stress.