On the development of an unstructured grid solver for inert and reactive high speed flow simulations

An unstructured grid Euler solver for reactive compressible flow applications is presented. The method is implemented in a cell centered, finite volume context for unstructured triangular grids. Three different schemes for spatial discretization are implemented and analyzed. Time march is implemented in a time-split fashion with independent integrators for the flow and chemistry equations. The capability implemented is tested for inert flows in a hypersonic inlet and for inert and reactive supersonic flows over a 2-D wedge. The results of the different schemes are compared with each other and with independent calculations using a structured grid code. The strengths and the possible weaknesses of the proposed methods are discussed.

Nitrogen doses and weed control via intercropping with gliricidia for corn production

Many studies have demonstrated the beneficial influence of nitrogen doses on corn dry grain yield and green ear yield. Due to a growing concern with environmental degradation, many agricultural practices, adopted in the past, are being reexamined. With regard to weed control, strategies that employ mechanical control, including intercrops, are being the object of renewed interest. The purpose of this study was to evaluate the effects of the application of nitrogen doses (0, 40, 80, and 120 kg N ha-1; as ammonium sulfate) and weed control on the growth, green ear yield, and grain yield of the AG 1051 corn cultivar. A randomized block experimental design with split-plots and nine replications was adopted. In addition to nitrogen rates, the AG 1051 cultivar was submitted to the following treatments, applied to subplots: no weeding, two hoeings (at 20 and 40 days after sowing), and intercropping with gliricídia (Gliricidia sepium). Gliricidia was sowed at corn planting, between the corn rows, using two seedlings per pit, in pits spaced 0.30 m apart. Gliricidia did not provide weed control, and gave plant growth, green ear yield and grain yield values similar to the no weeding treatment. However, regarding the number of mature ears got, intercropping with gliricidia did not differ from the two-hoeing treatment. Weed control did not have an effect on plant height and number of marketable, husked green ears, with the application of 120 kg N ha-1; indicating that nitrogen improved the corn's competitive ability. The two-hoeing treatment provided the best means for total green ears weight, number of marketable husked ears, both unhusked and husked marketable ear weight, grain yield and its components than the other treatments. Nitrogen application increased corn growth, green ear yield, and grain yield, as well as weed green biomass, but reduced the stand and growth of gliricidia.

Leaf contents of nitrogen and phenolic compounds and their bearing with the herbivore damage to Tibouchina pulchra Cogn. (Melastomataceae), under the influence of air pollutants from industries of Cubatão, São Paulo

The Atlantic Forest on the slopes of Serra do Mar around Cubatão (São Paulo, Brazil) has been affected by massive emissions of pollutants from the local growing industrial complex. The effects of air pollution on the amounts of leaf nitrogen, total soluble phenols and total tannins of Tibouchina pulchra Cogn., a common species in the area of Cubatão, were investigated, as well as the possible influence of the altered parameters on the leaf area damaged by herbivores. Fully expanded leaves were collected at two sites: the valley of Pilões river (VP), characterized by a vegetation virtually not affected by air pollution and taken as a reference; and valley of Mogi river (VM), close to the core region of the industrial complex, and severely affected by air pollution. No differences were observed for any parameters between samples collected in the summer and winter in both sites. On the other hand, compared to VP, individuals growing in VM presented higher amounts of nitrogen and lower amounts of total soluble phenols and total tannins, as well as higher percentages of galls per leaf and higher leaf area lost to herbivores. Regression analysis revealed that the increase in leaf area lost to herbivores can be explained by the increase of the content of nitrogen and decrease in the contents of total soluble phenols and total tannins. Although significant, the coefficients of explanation found were low for all analyses, suggesting that other biotic or abiotic factors are likely to influence leaf attack by herbivores.

Effects of different ammoniacal nitrogen sources on N-metabolism of the atmospheric bromeliad Tillandsia pohliana Mez

Increasing levels of atmospheric ammonia from anthropogenic sources have become a serious problem for natural vegetation. Short-term effects of different ammoniacal sources on the N metabolism of Tillandsia pohliana, an atmospheric bromeliad, were investigated. One-year-old, aseptically grown plants were transferred to a modified Knudson medium lacking N for three weeks. Plants were subsequently transferred to Knudson media supplemented with 0.5, 1.0, or 1.5 mM of N in the forms of NH3 or NH4+ as the sole N source. The activities of glutamine synthetase (GS) and glutamate dehydrogenase (GDH-NADH) were determined after 40 h. The GS activity was stimulated significantly by increasing the levels of the gaseous form. The GDH-NADH activity increased significantly under increasing N concentrations with NH3, while no significant differences were observed with NH4+ as a N source. These results may reflect a faster NH3 absorption by T. pohliana compared to NH4+ uptake. The increased activity of GDH-NADH in NH3 treatment may play a role in protecting the cells from the toxic effects of increased endogenous level of free ammonium. A raise in the concentration of N, especially in the form of NH3, greatly increased the content of free amino acids and soluble proteins. A possible utilisation of T. pohliana to evaluate the changes of atmospheric gaseous ammonia is proposed.

Fructan production in Vernonia herbacea (Vell.) Rusby is related to adequate nitrogen supply and period of cultivation

Previous studies showed that plants of Vernonia herbacea grown for one year under a limited nitrogen supply presented reduced growth and higher fructan content than plants treated with sufficient nitrogen supply. However, the total fructan production was similar in both plant groups due to the higher biomass of the underground reserve organ in nitrogen-sufficient (N-sufficient) plants. In the present study we aimed to evaluate if a stress growing condition under nitrogen-limited (N-limited) supply, following cultivation under N-sufficient supply would have a positive effect on fructan production. Plants cultivated during one year under N-sufficient supply (10.7 mmol L-1 N-NO3-) were separated in two groups. During the following six months, one group continued to receive the same treatment (control) while the other received an N-limited supply (1.3 mmol L-1 N-NO3-). Growth, photosynthesis and soluble carbohydrates were measured at days 0, 30, 60, 90 and 180. At day 30, plants transferred to N-limited supply showed a significant increase in growth and a decrease in fructan concentration, as a response to the stressing condition. However, in the following period growth was reduced and fructan concentration was increased, confirming the inverse relationship between nitrogen concentration and fructan content. After 180 days, although the fructan concentration in N-limited was significantly higher, with a fructan production of 6.0 g plant¹, the higher gain in rhizophore biomass after 18 months of cultivation in N-sufficient solution led to a fructan production of 8.3 g plant¹, thus surpassing the higher fructan concentration of N-limited plants.

Variation in nitrogen use strategies and photosynthetic pathways among vascular epiphytes in the Brazilian Central Amazon

The variation in nitrogen use strategies and photosynthetic pathways among vascular epiphyte families was addressed in a white-sand vegetation in the Brazilian Central Amazon. Foliar nitrogen and carbon concentrations and their isotopic composition (δ15N and δ13C, respectively) were measured in epiphytes (Araceae, Bromeliaceae and Orchidaceae) and their host trees. The host tree Aldina heterophylla had higher foliar N concentration and lower C:N ratio (2.1 ± 0.06% and 23.6 ± 0.8) than its dwellers. Tree foliar δ15N differed only from that of the orchids. Comparing the epiphyte families, the aroids had the highest foliar N concentration and lowest C:N ratios (1.4 ± 0.1% and 34.9 ± 4.2, respectively). The orchids had more negative foliar δ15N values (-3.5 ± 0.2‰) than the aroids (-1.9 ± 0.7‰) and the bromeliads (-1.1 ± 0.6‰). Within each family, aroid and orchid taxa differed in relation to foliar N concentrations and C:N ratios, whereas no internal variation was detected within bromeliads. The differences in foliar δ15N observed herein seem to be related to the differential reliance on the available N sources for epiphytes, as well as to the microhabitat quality within the canopy. In relation to epiphyte foliar δ13C, the majority of epiphytes use the water-conserving CAM-pathway (δ13C values around -17‰), commonly associated with plants that live under limited and intermittent water supply. Only the aroids and one orchid taxon indicated the use of C3-pathway (δ13C values around -30‰).

Concanavalin A-reactive nuclear matrix glycoprotein

The binding capacity of concanavalin A (Con A) to condensed euchromatin and heterochromatin was investigated in chicken erythrocyte nuclei (CEN), mouse liver cells, Zea mays mays meristematic cells and Drosophila melanogaster polytene chromosomes after 4 N HCl hydrolysis to determine whether binding was preferentially occurring in bands and heterochromatin. Dry mass (DM) variation was investigated in CEN by interference microscopy. Feulgen and Con A reactions were employed for all materials to correlate the loci of the two reactions. Quantifications and topological verifications were carried out by video image analysis (high performance cytometry). It was observed that 4 N HCl hydrolysis caused an important DM loss in CEN leaving a level corresponding to the average DNA DM content. In this material, Con A binding was restricted to the nuclear envelope, which reinforces the idea of the absence of a nuclear matrix in these cells. The other cell types exhibited a correspondence of Feulgen-positive and Con A-reactive areas. The Con A reaction was highly positive in the condensed chromatin areas and heterochromatin. This fact led us to speculate that Con A-positive proteins may play a role in the chromatin condensation mechanism, endowing this structure with physico-chemical stability towards acid hydrolysis and contributing to its rheological properties.

Levels of C-reactive protein in serum samples from healthy children and adults in São Paulo, Brazil

C-reactive protein (CRP) was measured by ELISA in the sera of 165 healthy blood donors and 125 normal children 1 to 14 years old. The serum levels of blood donors ranged from 0.05 to 57.6 mg/l with median and mean values of 1.8 mg/l and 4.86 mg/l, respectively. CRP levels ranged from 0.02 to 14.4 mg/l in the children's sera, the median being 0.45 mg/l and the mean 1.65 mg/l. No individual lacking CRP was detected. The high CRP levels observed in the present study suggest that the population of the State of São Paulo may usually be exposed to subacute infections and/or inflammation without presenting clinical symptoms

Precipitated immune complexes of IgM induce the generation of reactive oxygen species by rabbit polymorphonuclear leucocytes

We report that immune complexes of IgM (ICIgM) antibodies and ovalbumin in the form of a precipitate from the equivalence zone induce the generation of reactive oxygen species by rabbit blood polymorphonuclear leucocytes (PMN), as measured by the chemiluminescence (CL) production in the presence of luminol. The kinetics of CL generation induced by ICIgM is quite different from that induced by precipitated immune complexes of IgG (ICIgG): the maximum rate of CL production for ICIgM occurs around 14 min, whereas for ICIgG it occurs about 5 min after incubation with the cells. Also the triggering of the process requires a higher concentration of ICIgM than of ICIgG. Evidence is presented that these effects are not mediated by interaction of the antigen (ovalbumin) with the cell, since immune precipitates of ovalbumin and the F(ab')2 fragment had no effect. Our observations that precipitated ICIgM can also be an effective stimulus for CL generation and thus for O2- production reveal a new functional capability of PMN. These results may have implications for the understanding of the participation of ICIgM (as well as of ICIgG) in inflammatory reactions mediated by PMN in immune complex diseases, and in the mechanisms of defense against microbes and other non-self agents.

Purification and binding analysis of the nitrogen fixation regulatory NifA protein from Azospirillum brasilense

NifA protein activates transcription of nitrogen fixation operons by the alternative sigma54 holoenzyme form of RNA polymerase. This protein binds to a well-defined upstream activator sequence (UAS) located at the -200/-100 position of nif promoters with the consensus motif TGT-N10-ACA. NifA of Azospirillum brasilense was purified in the form of a glutathione-S-transferase (GST)-NifA fusion protein and proteolytic release of GST yielded inactive and partially soluble NifA. However, the purified NifA was able to induce the production of specific anti-A. brasilense NifA-antiserum that recognized NifA from A. brasilense but not from K. pneumoniae. Both GST-NifA and NifA expressed from the E. coli tac promoter are able to activate transcription from the nifHDK promoter but only in an A. brasilense background. In order to investigate the mechanism that regulates NifA binding capacity we have used E. coli total protein extracts expressing A. brasilense nifA in mobility shift assays. DNA fragments carrying the two overlapping, wild-type or mutated UAS motifs present in the nifH promoter region revealed a retarded band of related size. These data show that the binding activity present in the C-terminal domain of A. brasilense NifA protein is still functional even in the presence of oxygen.

Cytotoxic activity of BCG-activated macrophages against L929 tumor cells is nitric oxide-dependent

The tumoricidal activity of activated macrophages has been attributed largely to the release of tumor necrosis factor (TNF), or to the production of reactive oxygen or nitrogen intermediates. The L929 tumor cell line (a murine fibroblast-like cell) when treated with actinomycin D (ActD) has been used to measure TNFa cytotoxicity. In the present study, we determined the cytotoxic activity of BCG-activated peritoneal macrophages against ActD-untreated L929 tumor cells. Furthermore, we measured the production of hydrogen peroxide (H2O2), nitric oxide (NO) and TNF by macrophages cultured in the presence or absence of L929 cells. As expected, BCG-activated macrophages produced significant amounts of H2O2 (16.0 ± 3.0 µM), TNF (512 U/ml) and NO (71.5 ± 3.2 µM). TNF (256 U/ml) and NO (78.9 ± 9.7 µM) production was unchanged in co-cultures of L929 cells with BCG-activated macrophages but H2O2 production was totally inhibited. The cytotoxic activity was dependent on NO release since L-NAME (2.5, 5.0 and 10 mM), which blocks NO synthase, inhibited the killing of L929 cells. Addition of anti-TNF (20 µg/ml) antibodies to the cultures did not affect the tumoricidal activity of macrophages. Our results indicate that macrophage-mediated killing of L929 cells is largely dependent on NO production but independent of H2O2 or TNF release.

The antioxidant action of Polypodium leucotomos extract and kojic acid: reactions with reactive oxygen species

Two natural products Polypodium leucotomos extract (PL) and kojic acid (KA) were tested for their ability to scavenge reactive oxygen species (·OH, ·O2-, H2O2, ¹O2) in phosphate buffer. Hydroxyl radicals were generated by the Fenton reaction, and the rate constants of scavenging were 1.6 x 10(9) M-1 s-1 for KA and 1.0 x 10(9) M-1 s-1 for PL, similar to that of ethanol (1.4 x 10(9) M-1 s-1). With superoxide anions generated by the xanthine/hypoxanthine system, KA and PL (0.2-1.0 mg/ml) inhibited ·O2-dependent reduction of nitroblue tetrazolium by up to 30 and 31%, respectively. In the detection of ¹O2 by rose bengal irradiation, PL at 1.0 mg/ml quenched singlet oxygen by 43% relative to azide and KA by 36%. The present study demonstrates that PL showed an antioxidant effect, scavenging three of four reactive oxygen species tested here. Unlike KA, PL did not significantly scavenge hydrogen peroxide.

Different biochemical strategies of two Neotropical fish to cope with the impairment of nitrogen excretion during air exposure

The exposure of fish to air is normally expected to interfere with the nitrogen excretion process. Hoplias malabaricus and Hoplerythrinus unitaeniatus, two teleost species, display distinct behaviors in response to decreases in natural reservoir water levels, although they may employ similar biochemical strategies. To investigate this point, plasma levels of ammonia, urea, uric acid, and the two urea cycle enzymes, ornithine carbamoyl transferase (OCT) and arginase (ARG), as well as glutamine synthetase (GS) were determined for both species after exposure to air. Plasma ammonia increased gradually during exposure to air, but only H. malabaricus showed increased concentrations of urea. Plasma uric acid remained very low in both fish. Enzymatic activities (mean ± SD, µmol min-1 g protein-1) of H. malabaricus showed significant increases (P<0.05, N = 6) in OCT from 0.84 ± 0.05 to 1.42 ± 0.03, in ARG from 8.07 ± 0.47 to 9.97 ± 0.53 and in GS from 1.15 ± 0.03 to 2.39 ± 0.04. The OCT and ARG enzymes remained constant in H. unitaeniatus (N = 6), but GS increased from 1.49 ± 0.02 to 2.06 ± 0.03. Although these species are very closely related and share the same environment, their biochemical strategies in response to exposure to air or to increased plasma ammonia are different.

A method for multiple sequential analyses of macrophage functions using a small single cell sample

Microbial pathogens such as bacillus Calmette-Guérin (BCG) induce the activation of macrophages. Activated macrophages can be characterized by the increased production of reactive oxygen and nitrogen metabolites, generated via NADPH oxidase and inducible nitric oxide synthase, respectively, and by the increased expression of major histocompatibility complex class II molecules (MHC II). Multiple microassays have been developed to measure these parameters. Usually each assay requires 2-5 x 10(5) cells per well. In some experimental conditions the number of cells is the limiting factor for the phenotypic characterization of macrophages. Here we describe a method whereby this limitation can be circumvented. Using a single 96-well microassay and a very small number of peritoneal cells obtained from C3H/HePas mice, containing as little as <=2 x 10(5) macrophages per well, we determined sequentially the oxidative burst (H2O2), nitric oxide production and MHC II (IAk) expression of BCG-activated macrophages. More specifically, with 100 µl of cell suspension it was possible to quantify H2O2 release and nitric oxide production after 1 and 48 h, respectively, and IAk expression after 48 h of cell culture. In addition, this microassay is easy to perform, highly reproducible and more economical.

The role of complement in the modulation by fluid-phase IgG of the production of reactive oxygen species by polymorphonuclear leukocytes stimulated with IgG immune complexes

The production of reactive oxygen species (ROS) by polymorphonuclear leukocytes (PMN) can be induced by immune complexes and is an important component of phagocytosis in the killing of microorganisms, but can also be involved in inflammatory reactions when immune complexes are deposited in tissues. We have observed that fluid-phase IgG can inhibit the generation of ROS by rabbit PMN stimulated with precipitated immune complexes of IgG (ICIgG) in a dose-dependent manner, acting as a modulatory factor in the range of physiological IgG concentrations. This inhibitory effect is compatible with the known affinity (Kd) of monomeric IgG for the receptors involved (FcRII and FcRIII). The presence of complement components in the immune complexes results in a higher stimulation of ROS production. In this case, however, there is no inhibition by fluid-phase IgG. The effect of complement is strongly dependent on the presence of divalent cations (Ca2+ or Mg2+) in the medium, whereas the stimulation of ICIgG (without complement) does not depend on these cations. We have obtained some evidence indicating that iC3b should be the component involved in the effect of complement through interaction with the CR3 receptor. The absence of the inhibitory effect of fluid-phase IgG in ROS production when complement is present in the immune complex shows that complement may be important in vivo not only in the production of chemotactic factors for PMN, but also in the next phase of the process, i.e., the generation of ROS.