A multi-level analytical approach for detection and visualization of intracellular NO production and nitrosation events using diaminofluoresceins

Diaminofluoresceins are widely used probes for detection and intracellular localization of NO formation in cultured/isolated cells and intact tissues. The fluorinated derivative, 4-amino-5-methylamino-2′,7′-difluorofluorescein (DAF-FM), has gained increasing popularity in recent years due to its improved NO-sensitivity, pH-stability, and resistance to photo-bleaching compared to the first-generation compound, DAF-2. Detection of NO production by either reagent relies on conversion of the parent compound into a fluorescent triazole, DAF-FM-T and DAF-2-T, respectively. While this reaction is specific for NO and/or reactive nitrosating species, it is also affected by the presence of oxidants/antioxidants. Moreover, the reaction with other molecules can lead to the formation of fluorescent products other than the expected triazole. Thus additional controls and structural confirmation of the reaction products are essential. Using human red blood cells as an exemplary cellular system we here describe robust protocols for the analysis of intracellular DAF-FM-T formation using an array of fluorescence-based methods (laser-scanning fluorescence microscopy, flow cytometry and fluorimetry) and analytical separation techniques (reversed-phase HPLC and LC-MS/MS). When used in combination, these assays afford unequivocal identification of the fluorescent signal as being derived from NO and are applicable to most other cellular systems without or with only minor modifications.

A framework for designing multi-functional cover crops

Cover crops are sown to provide a number of ecosystem services including nutrient management, mitigation of diffuse pollution, improving soil structure and organic matter content, weed suppression, nitrogen fixation and provision of resources for biodiversity. Although the decision to sow a cover crop may be driven by a desire to achieve just one of these objectives, the diversity of cover crops species and mixtures available means that there is potential to combine a number of ecosystem services within the same crop and growing season. Designing multi-functional cover crops would potentially help to reconcile the often conflicting agronomic and environmental agendas and contribute to the optimal use of land. We present a framework for integrating multiple ecosystem services delivered by cover crops that aims to design a mixture of species with complementary growth habit and functionality. The optimal number and identity of species will depend on the services included in the analysis, the functional space represented by the available species pool and the community dynamics of the crop in terms of dominance and co-existence. Experience from a project that applied the framework to fertility building leys in organic systems demonstrated its potential and emphasised the importance of the initial choice of species to include in the analysis

The challenge of maintaining Atlantic forest biodiversity: A multi-taxa conservation assessment of specialist and generalist species in an agro-forestry mosaic in southern Bahia

Recent developments have highlighted the importance of forest amount at large spatial scales and of matrix quality for ecological processes in remnants. These developments, in turn, suggest the potential for reducing biodiversity loss through the maintenance of a high percentage of forest combined with sensitive management of anthropogenic areas. We conducted a multi-taxa survey to evaluate the potential for biodiversity maintenance in an Atlantic forest landscape that presented a favorable context from a theoretical perspective (high proportion of mature forest partly surrounded by structurally complex matrices). We sampled ferns, butterflies, frogs, lizards, bats, small mammals and birds in interiors and edges of large and small mature forest remnants and two matrices (second-growth forests and shade cacao plantations), as well as trees in interiors of small and large remnants. By considering richness, abundance and composition of forest specialists and generalists, we investigated the biodiversity value of matrix habitats (comparing them with interiors of large remnants for all groups except tree), and evaluated area (for all groups) and edge effects (for all groups except trees) in mature forest remnants. our results suggest that in landscapes comprising high amounts of mature forest and low contrasting matrices: (1) shade cacao plantations and second-growth forests harbor an appreciable number of forest specialists; (2) most forest specialist assemblages are not affected by area or edge effects, while most generalist assemblages proliferate at edges of small remnants. Nevertheless, differences in tree assemblages, especially among smaller trees, Suggest that observed patterns are unlikely to be stable over time. (C) 2009 Elsevier Ltd. All rights reserved.

Development of a new analytical approach based on cellulose membrane and chelator for differentiation of labile and inert metal species in aquatic systems

A new procedure was developed in this study, based on a system equipped with a cellulose membrane and a tetraethylenepentamine hexaacetate chelator (MD-TEPHA) for in situ characterization of the lability of metal species in aquatic systems. To this end, the DM-TEPHA system was prepared by adding TEPHA chelator to cellulose bags pre-purified with 1.0 mol L-1 of HCl and NaOH solutions. After the MD-TEPHA system was sealed, it was examined in the laboratory to evaluate the influence of complexation time (0-24 h), pH (3.0, 4.0, 5.0, 6.0 and 7.0), metal ions (Cu, Cd, Fe, Mn and Ni) and concentration of organic matter (15, 30 and 60 mg L-1) on the relative lability of metal species by TEPHA chelator. The results showed that Fe and Cu metals were complexed more slowly by TEPHA chelator in the MD-TEPHA system than were Cd, Ni and Mn in all pH used. It was also found that the pH strongly influences the process of metal complexation by the MD-TEPHA system. At all the pH levels, Cd, Mn and Ni showed greater complexation with TEPHA chelator (recovery of about 95-75%) than did Cu and Fe metals. Time also affects the lability of metal species complexed by aquatic humic substances (AHS); while Cd, Ni and Mn showed a faster kinetics, reaching equilibrium after about 100 min, and Cu and Fe approached equilibrium after 400 min. Increasing the AHS concentration decreases the lability of metal species by shifting the equilibrium to AHS-metal complexes. Our results indicate that the system under study offers an interesting alternative that can be applied to in situ experiments for differentiation of labile and inert metal species in aquatic systems. (c) 2006 Elsevier B.V. All rights reserved.

Neural network based on adaptive resonance theory with continuous training for multi-configuration transient stability analysis of electric power systems

This work presents a methodology to analyze electric power systems transient stability for first swing using a neural network based on adaptive resonance theory (ART) architecture, called Euclidean ARTMAP neural network. The ART architectures present plasticity and stability characteristics, which are very important for the training and to execute the analysis in a fast way. The Euclidean ARTMAP version provides more accurate and faster solutions, when compared to the fuzzy ARTMAP configuration. Three steps are necessary for the network working, training, analysis and continuous training. The training step requires much effort (processing) while the analysis is effectuated almost without computational effort. The proposed network allows approaching several topologies of the electric system at the same time; therefore it is an alternative for real time transient stability of electric power systems. To illustrate the proposed neural network an application is presented for a multi-machine electric power systems composed of 10 synchronous machines, 45 buses and 73 transmission lines. (C) 2010 Elsevier B.V. All rights reserved.

In situ application of a cellulose bag and an ion exchanger for differentiation of labile and inert metal species in aquatic systems

This work involved the development and application of a new analytical procedure for in-situ characterization of the lability of metal species in aquatic systems by using a system equipped with a diffusion membrane and cellulose organomodified with p-aminobenzoic acid groups (DM-Cell-PAB). To this end, the DM-Cell-PAB system was prepared by adding cellulose organomodified with p-aminobenzoic acid groups (Cell-PAB) to pre-purified cellulose bags. After the DM-Cell-PAB system was sealed, it was examined in the laboratory. The in-situ application involved immersing the DM-Cell-PAB system in two different rivers, enabling us to study the relative lability of metal species (Cu, Cd, Fe, Mn, and Ni) as a function of time and quantity of exchanger. The procedure is simple and opens up a new perspective for understanding environmental phenomena relating to the complexation, transport, stability, and lability of metal species in aquatic systems rich in organic matter.

Morphology and histology of male and female reproductive systems in the inseminating species Scoloplax distolothrix (Ostariophysi : Siluriformes : Scoloplacidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

New analytical procedure based on a cellulose bag and ionic exchanger with p-aminobenzoic acid groups for differentiation of labile and inert metal species in aquatic systems

A new procedure was developed for the in situ characterization of the lability of metal species in aquatic systems by using a system equipped with a diffusion membrane and cellulose organomodified with p-aminobenzoic acid groups (DM-Cell-PAB). To this end, the DM-Cell-PAB system was prepared by adding cellulose organomodified with p-aminobenzoic acid groups (Cell-PAB) to pre-purified cellulose bags. After the DM-Cell-PAB system was sealed, it was examined in the laboratory to evaluate the influence of complexation time, mass of exchanger, pH, metal ions (Cu, Cd, Fe, Mn, and Ni), and concentration of organic matter on the relative lability of metal species. It was found that the pH and kinetics strongly influence the process of metal complexation by the DM-Cell-PAB system. At all pH levels, Cd, Mn, and Ni showed lower complexation with Cell-PAB resin than Cu and Fe metals. Note that relative lability of metals complexed to aquatic humic substances (AHS) in the presence of Cell-PAB resin showed the following order: Cu congruent to Fe >> Ni > Mn=Cd. The results presented here also indicate that increasing the AHS concentration decreases the lability of metal species by shifting the equilibrium to AHS-metal complexes. Our results indicate that the system under study offers an interesting alternative that can be applied to in situ experiments for differentiation of labile and inert metal species in aquatic systems.

Multi-spectroscopic monitoring of cisplatin-derived species delivery from ureasil polyether hybrid matrix

Driven by the challenges involved in the development of new advanced materials with unusual drug delivery profiles capable of improving the therapeutic and toxicological properties of existing cancer chemotherapy, the one-pot sol-gel synthesis of flexible, transparent and insoluble urea-cross-linked polyether-siloxane hybrids has been recently developed. In this one-pot synthesis, the strong interaction between the antitumor cisplatin (CisPt) molecules and the ureasil-poly(propylene oxide) (PPO) hybrid matrix gives rise to the incorporation and release of an unknown CisPt-derived species, hindering the quantitative determination of the drug release pattern from the conventional UV-Vis absorption technique. In this article, we report the use of an original synchrotron radiation calibration method based on the combination of XAS and UV-Vis for the quantitative determination of the amount of Pt-based molecules released in water. Thanks to the combination of UV-Vis, XAS and Raman techniques, we demonstrated that both the CisPt molecules and the CisPt-derived species are loaded into an ureasil-PPO/ureasil-poly(ethylene oxide) (PEO) hybrid blend matrix. The experimentally determined molar extinction coefficient of the CisPt-derived species loaded into ureasil-PPO hybrid matrix enabled the simultaneous time-resolved monitoring of each Pt species released from this hybrid blend matrix.

Minimization of lead and copper interferences on spectrophotometric determination of cadmium using electrolytic deposition and ion-exchange in multi-commutation flow system

A new strategy for minimization of Cu2+ and Pb2+ interferences on the spectrophotometric determination of Cd2+ by the Malachite green (MG)-iodide reaction using electrolytic deposition of interfering species and solid phase extraction of Cd2+ in flow system is proposed. The electrolytic cell comprises two coiled Pt electrodes concentrically assembled. When the sample solution is electrolyzed in a mixed solution containing 5% (v/v) HNO3, 0.1% (v/v) H2SO4 and 0.5 M NaCl, Cu2+ is deposited as Cu on the cathode, Pb2+ is deposited as PbO2 on the anode while Cd2+ is kept in solution. After electrolysis, the remaining solution passes through an AG1-X8 resin (chloride form) packed minicolumn in which Cd2+ is extracted as CdCl4/2-. Electrolyte compositions, flow rates, timing, applied current, and electrolysis time was investigated. With 60 s electrolysis time, 0.25 A applied current, Pb2+ and Cu2+ levels up to 50 and 250 mg 1-1, respectively, can be tolerated without interference. For 90 s resin loading time, a linear relationship between absorbance and analyte concentration in the 5.00-50.0 μg Cd 1-1 range (r2 = 0.9996) is obtained. A throughput of 20 samples per h is achieved, corresponding to about 0.7 mg MG and 500 mg KI and 5 ml sample consumed per determination. The detection limit is 0.23 μg Cd 1-1. The accuracy was checked for cadmium determination in standard reference materials, vegetables and tap water. Results were in agreement with certified values of standard reference materials and with those obtained by graphite furnace atomic absorption spectrometry at 95% confidence level. The R.S.D. for plant digests and water containing 13.0 μg Cd 1-1 was 3.85% (n = 12). The recoveries of analyte spikes added to the water and vegetable samples ranged from 94 to 104%. (C) 2000 Elsevier Science B.V.

Growth parameters and nutrient content in four multipurpose tree species with potential characteristics for agroforestry systems in a Cerrado region in Botucatu, São Paulo State, Brazil

In order to evaluate growth characteristics, adaptability, biomass production, nutrient recycling, nutrient distribution and the ability to regenerate degraded land, a trial using four multipurpose tree species (Leucaena leucocephala, Leucaena diversifolia, Acacia melanoxylon and Mimosa scabrella) was undertaken over two years in a distrophic red yellow latosol (oxisol) following a randomized block experimental design with four replications. At the age of two years, A. melanoxylon and L. diversifolia were the tallest species (5.25 and 4.97 m, respectively) and A. melanoxylon and M. scabrella had the largest diameters at 20 cm from tree base. Mimosa scabrella and A. melanoxylon had the highest dry matter production and quantity of nutrients in the above ground biomass. In all species, the highest nutrient contents were found in the leaves, followed by branches and stems. From all species, the highest Nutrient Utilization Efficiency Indexes were obtained for sulphur, phosphorous, and magnesium; L. diversifolia was the most efficient for nitrogen, potassium, calcium, sulphur, and manganese, while A. melanoxylon was the most efficient for phosphorus, magnesium, boron, iron, and zinc. Litter production levels over a three month period were as follows: M. scabrella > A. melanoxylon > L. diversifolia > L. leucocephala. Litter nutrient content was higher in M. scabrella than in the other species.

Plant species identification using multi-scale fractal dimension applied to images of adaxial surface epidermis

This paper presents the study of computational methods applied to histological texture analysis in order to identify plant species, a very difficult task due to the great similarity among some species and presence of irregularities in a given species. Experiments were performed considering 300 ×300 texture windows extracted from adaxial surface epidermis from eight species. Different texture methods were evaluated using Linear Discriminant Analysis (LDA). Results showed that methods based on complexity analysis perform a better texture discrimination, so conducting to a more accurate identification of plant species. © 2009 Springer Berlin Heidelberg.

Multi-dimensional effects on Cladoceran (Crustacea, Anomopoda) assemblages in two cascade reservoirs in Southeast Brazil

The Cladocera assemblages in two cascade reservoirs located in the Paranapanema River in Brazil were studied during two consecutive years. Upstream Chavantes Reservoir is an accumulation system, with a long water retention time, high depth and oligo-mesotrophic status. The downstream Salto Grande Reservoir is a small, run-of-river reservoir, with a short water retention time, shallow depth and meso-eutrophic status. The goal of this study was to determine the inter- and intra-reservoir limnological differences with emphasis on the Cladocerans assemblages. The following questions were posed: (i) what are the seasonal dynamics of the reservoir spatial structures; (ii) how dynamics, seasonally, is the reservoirs spatial structure; and (iii) are the reservoir independent systems? A total of 43 Cladoceran species were identified in this study. Ceriodaphnia silvestrii was the most abundant and frequent species found in Chavantes Reservoir, while C. cornuta was most abundant and frequent in Salto Grande Reservoir. The Cladoceran species richness differed significantly among sampling sites for both reservoirs. In terms of abundance, there was a significant variation among sampling sites and periods for both reservoirs. A cluster analysis indicated a higher similarity among the deeper compartments, and the intermediate river-reservoir zones was grouped with the riverine sampling sites. For the smaller Salto Grande Reservoir, the entrance of a middle size tributary causes major changes in the system. A distinct environment was observed in the river mouth zone of another small tributary, representing a shallow environment with aquatic macrophyte stands. A canonical correlation analysis between environmental variables and Cladoceran abundance explained 75% of the data variability, and a complementary factorial analysis explained 65% of the variability. The spatial compartmentalization of the reservoirs, as well as the particular characteristics of the two study reservoirs, directly influenced the structure of the Cladoceran assemblages. The conditions of the lacustrine (dam) zone of the larger Chavantes Reservoir were reflected in the upstream zone of the smaller downstream Salto Grande Reservoir, highlighting the importance of plankton exportation in reservoir cascade systems. The comparative spatial-temporal analysis indicated conspicuous differences between the two reservoirs, reinforcing the necessity of considering tropical/subtropical reservoirs as complex, multi-compartmental water systems. © 2010 The Authors. Journal compilation © 2010 Blackwell Publishing Asia Pty Ltd.

Eimeria species in young and adult sheep raised under intensive and/or semi-intensive systems of a herd from Umuarama city, Parana State, Brazil

The present study aimed to identify Eimeria species in young and adult sheep raised under intensive and / or semi-intensive systems of a herd from Umuarama city, Parana State, Brazil using the traditional diagnostic methods and to correlate the infection level/types of infection in the different age/system in this herd. Fecal samples were collected from the rectum of 210 sheep and were subjected to laboratory analysis to differentiate the species. Furthermore, animals were observed to determine the occurrences of the clinical or subclinical forms of eimeriosis. Out of the 210 collected fecal samples, 147 (70%) were positive for Eimeria oocysts, and 101 (47.86%) belonged to young animals that were raised under intensive and / or semi-intensive farming systems. Oocysts from 9 species of Eimeria parasites were identified in the sheep at the following prevalence rates: E. crandallis, 50.0%; E. parva, 21.6%; E. faurei, 8.1%; E. ahsata, 8.1%; E. intricata, 5.4%; E. granulosa, 2.7%; E. ovinoidalis, 2.0%; E. ovina, 1.3%; and E. bakuensis, 0.6%. There were no differences regarding the more frequent Eimeria species among the different ages of animals or between the different farming management systems. Based on these data, E. crandallis was the most prevalent, followed by E. parva and E. faurei species, regardless of the age. Higher parasitism was diagnosed in the young animals that were raised in a confinement regime, and the disease found in the herd was classified as subclinical. Further studies should be conducted in this herd, to verify if the eimeriosis subclinical can cause damage especially in young animals with a high level of infection.