Nitrogen metabolism in leaves of a tank epiphytic bromeliad: characterization of a spatial and functional division.

The leaf is considered the most important vegetative organ of tank epiphytic bromeliads due to its ability to absorb and assimilate nutrients. However, little is known about the physiological characteristics of nutrient uptake and assimilation. In order to better understand the mechanisms utilized by some tank epiphytic bromeliads to optimize the nitrogen acquisition and assimilation, a study was proposed to verify the existence of a differential capacity to assimilate nitrogen in different leaf portions. The experiments were conducted using young plants of Vriesea gigantea. A nutrient solution containing NO3⁻/NH4⁺ or urea as the sole nitrogen source was supplied to the tank of these plants and the activities of urease, nitrate reductase (NR), glutamine synthetase (GS) and glutamate dehydrogenase (NADH-GDH) were quantified in apical and basal leaf portions after 1, 3, 6, 9, 12, 24 and 48 h. The endogenous ammonium and urea contents were also analyzed. Independent of the nitrogen sources utilized, NR and urease activities were higher in the basal portions of leaves in all the period analyzed. On the contrary, GS and GDH activities were higher in apical part. It was also observed that the endogenous ammonium and urea had the highest contents detected in the basal region. These results suggest that the basal portion was preferentially involved in nitrate reduction and urea hydrolysis, while the apical region could be the main area responsible for ammonium assimilation through the action of GS and GDH activities. Moreover, it was possible to infer that ammonium may be transported from the base, to the apex of the leaves. In conclusion, it was suggested that a spatial and functional division in nitrogen absorption and NH4⁺ assimilation between basal and apical leaf areas exists, ensuring that the majority of nitrogen available inside the tank is quickly used by bromeliad's leaves.

Effects of atypical (risperidone) and typical (haloperidol) antipsychotic agents on astroglial functions.

Although classical and atypical antipsychotics may have different neurotoxic effects, their underlying mechanisms remain to be elucidated, especially regarding neuroglial function. In the present study, we compared the atypical antipsychotic risperidone (0.01-10 µM) with the typical antipsychotic haloperidol (0.01-10 µM) regarding different aspects such as glutamate uptake, glutamine synthetase (GS) activity, glutathione (GSH) content, and intracellular reactive oxygen species (ROS) production in C6 astroglial cells. Risperidone significantly increased glutamate uptake (up to 27%), GS activity (14%), and GSH content (up to 17%). In contrast, haloperidol was not able to change any of these glial functions. However, at concentration of 10 µM, haloperidol increased (12%) ROS production. Our data contribute to the clarification of different hypothesis concerning the putative neural responses after stimulus with different antipsychotics, and may establish important insights about how brain rewiring could be enhanced.

Effect of composted textile sludge on growth, nodulation and nitrogen fixation of soybean and cowpea.

The effect of composted textile sludge on growth, nodulation and nitrogen fixation of soybean and cowpea was evaluated in a greenhouse experiment. The compost was incorporated into soil at 0, 9.5, 19 and 38 t ha(-1) (bases upon the N requirement of the crops, i.e., 0, 50, 100 and 200 kg available N ha(-1)). Growth, nodulation and shoot accumulation of nitrogen were evaluated 36 and 63 days after plant emergence. Nodule glutamine synthetase (GS) activity and leghemoglobin content were evaluated 63 days after emergence. Composted textile sludge did not show negative effects on nodule number and weight, nodule GS activity and leghemoglobin content. Nitrogen accumulation in shoot dry matter in soybean and cowpea was higher than other treatments with application of 19 t ha(-1) of compost. Composting can be an alternate technology for the management of solid textile mill sludge. This study verifies that the composted textile sludge was not harmful to growth, nodulation and nitrogen fixation of soybean and cowpea.

Biochemical responses of two erythrinidae fish to environmental ammonia.

The non-ionized form of ammonia is very toxic to many aquatic species. It is especially important in several aspects of fish biology. A large range of organismal strategies for coping with environmental stressors is usually observed in living organisms. Among those, the responses for managing chemical stressors are well studied. The present work compares biochemical responses of two evolutionarily close species, Hoplias malabaricus and Hoplerythrinus unitaeniatus, exposed to environmental ammonia. Adult fish were submitted to 1.0 mg/L of ammonium chloride for 24 hours, and plasma ammonia and urea levels were determined. The activities of OUC enzymes OCT and ARG, and the accessory enzyme GS, were quantified in liver extract and are expressed below in mumol/min/mg of wet tissue. Increases in OUC enzymes (GS from 1.14 to 2.43, OCT from 0.81 to 1.72, and ARG from 3.15 to 4.23), plasma ammonia (from 0.95 to 1.42 mmol/L), and plasma urea (from 0.82 to 1.53 mmol/L) were observed (p < 0.05) in H. malabaricus exposed to 1 mg/L of ammonia chloride. The GS in H. unitaeniatus increased from 1.43 to 1.84, however the OCT, ARG, and plasma urea from H. unitaeniatus did not change. These data indicate that each species responds differently to the same environmental stressor.

Improved soybean root association of N-starved Bradyrhizobium japonicum.

In this study, we addressed the effects of N limitation in Bradyrhizobium japonicum for its association with soybean roots. The wild-type strain LP 3001 grew for six generations with a growth rate of 1.2 day(-1) in a minimal medium with 28 mM mannitol as the carbon source and with the N source [(NH(4))(2)SO(4)] limited to only 20 microM. Under these conditions, the glutamine synthetase (GS) activity was five to six times higher than in similar cultures grown with 1 or 0.1 mM (NH(4))(2)SO(4). The NtrBC-inducible GSII form of this enzyme accounted for 60% of the specific activity in N-starved rhizobia, being negligible in the other two cultures. The exopolysaccharide (EPS) and capsular polysaccharide (CPS) contents relative to cell protein were significantly higher in the N-starved cultures, but on the other hand, the poly-3-hydroxybutyrate level did not rise in comparison with N-sufficient cultures. In agreement with the accumulation of CPS in N-starved cultures, soybean lectin (SBL) binding as well as stimulation of rhizobial adsorption to soybean roots by SBL pretreatment were higher. The last effect was evident only in cultures that had not entered stationary phase. We also studied nodC gene induction in relation to N starvation. In the chromosomal nodC::lacZ fusion Bj110-573, nodC gene expression was induced by genistein 2.7-fold more in N-starved young cultures than in nonstarved ones. In stationary-phase cultures, nodC gene expression was similarly induced in N-limited cultures, but induction was negligible in cultures limited by another nutrient. Nodulation profiles obtained with strain LP 3001 grown under N starvation indicated that these cultures nodulated faster. In addition, as culture age increased, the nodulation efficiency decreased for two reasons: fewer nodules were formed, and nodulation was delayed. However, their relative importance was different according to the nutrient condition: in older cultures the overall decrease in the number of nodules was the main effect in N-starved cultures, whereas a delay in nodulation was more responsible for a loss in efficiency of N-sufficient cultures. Competition for nodulation was studied with young cultures of two wild-type strains differing only in their antibiotic resistance, the N-starved cultures being the most competitive.

Drought-Induced effects and recovery of nitrate assimilation and nodule activity in cowpea plants inoculated with Bradyrhizobium spp. under moderate nitrate level

This study was carried out to establish comparative effects of drought and recovery on the nitrate assimilation and nodule activity related to N2 fixation in cowpea plants [Vigna unguiculata L. (Walp.)] previously inoculated with Bradyrhizobium spp. BR-3256 (CB-756) strain in the presence of 5 mol m-3 NO-3. Twenty-eight-day-old nodulated plants were submitted to water deprivation during 4 consecutive days and afterwards resupplied with nutrient solution during 2 days. The water deprivation caused a rapid increase in the nitrate content in root and a marked reduction in leaf nitrate reductase (NR) activity. In contrast nodule NR activity was slightly increased by water deprivation. Concomitantly, in nodules of water stressed plants, leghemoglobin and glutamine synthetase (GS) activity declined and a progressive reduction in ureide-N concentration in xylem sap was observed. Leaf-NR activity increased rapidly after rehydration while leaf nitrate content declined. In contrast both GS activity and soluble protein content in the nodule continued to decline in rewatered plants. In addition the concentration of leghemoglobin recovered well, while the xylem ureide-N content experienced a slight increase after rehydration. Despite the nitrate assimilation in leaves and the nodule activity had been both severely affected by water stress, the rapid recovery of nitrate reductase activity suggests that the nitrate assimilation process is less sensitive to drought/rehydration cycle when cowpea plants are nodulated in presence of moderate nitrate level.

Water stress response on the enzymatic activity in cowpea nodules

A greenhouse experiment was carried out aiming to study the effect of water stress on metabolic activity of cowpea nodules at different plant development stages. Cowpea plants were grown in pots with yellow latosol soil under three different matric potentials treatments: -7.0 (control-S1), -70.0 (S2) and <-85.0 KPa (S3). The experimental design was randomized blocks with sub-divided plots, each plot containing a different degree of water stress, divided in sub-plots for the four different developmental stages: E1 (0-15), E2 (15-30), E3 (20-35) and E4 (30-45) days after emmergence. Water stress treatments were applied by monitoring soil water potential using a set of porous cups. The effect of water stress was most harmful to cowpea when it was applied at E2 than at other symbiotic process stages. Shoot/root ratio decreased from 2.61 to 2.14 when matric potential treatment was <-85.0 and -70.0 KPa respectively. There was a reduction in the glutamine synthetase activity and phosphoenolpyruvate carboxilase activity with increased stress, while glutamine synthase activity was the enzyme most sensitive to water stress. Glutamate dehydrogenase activity increased in more negative matric potential, indicating that this enzyme is sufficiently activitye under water stress.

Oxidative damage to lipids and DNA concurrent with decrease of antioxidants in rat testes after acute iron intoxication.

The effect of acute iron overload was studied in rat testes 20 h after a single administration of iron-dextran (500 mg/kg body wt, ip). Total testes iron content was 6.1-fold higher in iron-treated rats compared to controls. The endogenous level of lipid peroxidation was evaluated as 2-thiobarbituric acid-reactive substances (TBARS). Testes iron concentration (0.12-2.67 mumol/g of tissue) was positively correlated (r = 0.86; P < 0.01) with testes TBARS (26.2-77.5 nmol/g tissue). Testes content of lipid-soluble antioxidants, alpha-tocopherol, ubiquinol-9, and ubiquinol-10, were inversely correlated with testes iron content. The steady-state level of 8-oxo-7,8-dihydro-2'-deoxyguanosine in testes DNA was 25% higher (P < 0.01) in iron-treated rats compared to controls (2.4 +/- 0.2 oxo8dG/10(5)dG). The content of protein carbonyl groups (1.45 +/- 0.13 nmol/mg protein) and the activity of glutamine synthase (1.32 +/- 0.07 units/mg protein) were similar for the iron-treated and control rats. Fe treatment did not affect superoxide dismutase, catalase, and glutathione peroxidase activities. The results indicate that acute iron overload causes iron accumulation in rat testes, which is associated with increased lipid and DNA oxidative damage and depletion of lipid-soluble antioxidants.

Glutamine synthesis is a regulatory signal controlling glucose catabolism in Saccharomyces cerevisiae.

The effect of glutamine biosynthesis and degradation on glucose catabolism in Saccharomyces cerevisiae was studied. A wild-type strain and mutants altered in glutamine biosynthesis and degradation were analyzed. Cells having low levels of glutamine synthetase activity showed high ATP/ADP ratios and a diminished rate of glucose metabolism. It is proposed that glutamine biosynthesis plays a role in the regulation of glucose catabolism.

Nitrogen metabolism and chloramphenicol production in Streptomyces venezuelae.

The relationship between chloramphenicol production and nitrogen metabolism in Streptomyces venezuelae was examined in stirred jar cultures under pH control. Nitrogen sources that supported rapid biomass accumulation gave low rates of antibiotic synthesis during growth. This was consistent with a general incompatibility between fast growth and high yields of chloramphenicol. In media where the growth rate was reduced below the attainable maximum by the rate at which nitrogen could be assimilated, chloramphenicol production was associated with biomass accumulation. Enzymes that are potentially associated with nitrogen assimilation pathways were assayed in cultures supplied with nitrogen sources supporting markedly different growth rates. The results indicated that glutamine synthetase and alanine dehydrogenase levels were relatively insensitive to changes in growth rate and nitrogen source depletion. Glutamate dehydrogenase and glutamate synthase, on the other hand, showed high activity in cultures assimilating ammonium nitrogen and markedly decreased activity with poorer nitrogen sources or when ammonium was depleted. If chloramphenicol biosynthesis is coordinately controlled by mechanisms that regulate nitrogen assimilation, glutamate synthase and glutamate dehydrogenase are the most likely enzymes that manifest the regulatory linkage.