Regulation of nitrogen metabolism in salt tolerant and salt sensitive Frankia strains.

Effect of salinity (0, 50, 100, 250, 500 and 750 mM NaCl) was observed on some important physiological parameters of nitrogen metabolism such as nitrate uptake, intracellular and extracellular ammonium status and activities of nitrogenase, nitrate reductase, nitrite reductase and glutamine synthetase among Frankia strains differing in their salt tolerance capacity. Nitrogenase activity closely followed the growth pattern with regular decline on NaCl supplementation. All the other enzymes showed optimum activity at 100 mM and declined further. Co-regulation of the nitrate uptake system and sequential enzyme activities plays a crucial role in governing the nitrogen status of strains during salt stress. HsIi10 experiencing minimum decline in enzyme activities and best possible nitrogen regulation under NaCl replete condition showed adequate nutritional management. Among all the strains, HsIi10 proved to be salt tolerant on account of above features while the salt sensitive strain HsIi8 lacked the ability to regulate various steps of nitrogen metabolism during salinity, and thus Frankia strain HsIi10 can potentially serve as a potential biofertilizer in the saline soil.

beta-catenin activated hepatocellular adenoma

No abstract available.

Glial cell activity is maintained during prolonged inflammatory challenge in rats

We evaluated the expression of glial fibrillary acidic protein (GFAP), glutamine synthetase (GS), ionized calcium binding adaptor protein-1 (Iba-1), and ferritin in rats after single or repeated lipopolysaccharide (LPS) treatment, which is known to induce endotoxin tolerance and glial activation. Male Wistar rats (200-250 g) received ip injections of LPS (100 µg/kg) or saline for 6 days: 6 saline (N = 5), 5 saline + 1 LPS (N = 6) and 6 LPS (N = 6). After the sixth injection, the rats were perfused and the brains were collected for immunohistochemistry. After a single LPS dose, the number of GFAP-positive cells increased in the hypothalamic arcuate nucleus (ARC; 1 LPS: 35.6 ± 1.4 vs control: 23.1 ± 2.5) and hippocampus (1 LPS: 165.0 ± 3.0 vs control: 137.5 ± 2.5), and interestingly, 6 LPS injections further increased GFAP expression in these regions (ARC = 52.5 ± 4.3; hippocampus = 182.2 ± 4.1). We found a higher GS expression only in the hippocampus of the 6 LPS injections group (56.6 ± 0.8 vs 46.7 ± 1.9). Ferritin-positive cells increased similarly in the hippocampus of rats treated with a single (49.2 ± 1.7 vs 28.1 ± 1.9) or repeated (47.6 ± 1.1 vs 28.1 ± 1.9) LPS dose. Single LPS enhanced Iba-1 in the paraventricular nucleus (PVN: 92.8 ± 4.1 vs 65.2 ± 2.2) and hippocampus (99.4 ± 4.4 vs 73.8 ± 2.1), but had no effect in the retrochiasmatic nucleus (RCA) and ARC. Interestingly, 6 LPS increased the Iba-1 expression in these hypothalamic and hippocampal regions (RCA: 57.8 ± 4.6 vs 36.6 ± 2.2; ARC: 62.4 ± 6.0 vs 37.0 ± 2.2; PVN: 100.7 ± 4.4 vs 65.2 ± 2.2; hippocampus: 123.0 ± 3.8 vs 73.8 ± 2.1). The results suggest that repeated LPS treatment stimulates the expression of glial activation markers, protecting neuronal activity during prolonged inflammatory challenges.

Scavenging of nickel and chromium toxicity in Aulosira fertilissima by immobilization: effect on nitrogen assimilating enzymes

The ubiquity of heavy metals in the biosphere results in the introduction of high amounts of toxic metals into the food chain from various sources. In the present study, one of the strongest nitrogen fixing cyanobacterium of the rice fields, Aulosira fertilissima, was subjected to nickel and chromium stress and the ameliorating effect of immobilization was investigated. Cell immobilization could protect the organism's growth against the toxicity of both heavy metals at LC50 as compared to lethal concentrations. The nitrate reductase activity in free cells treated with the metals was substantially inhibited but immobilized cells treated with 0.1 ppm nickel was not affected by the metal treatment. Cell immobilization also resulted in a significant protection against sub-lethal concentration of chromium but to a lesser degree than it did with sub- lethal levels of nickel. Control immobilized cells also had higher Nitrogenase activity than control free cells. Nickel and chromium addition markedly decreased the enzyme activity in free cells but immobilized cells exposed to sublethal concentrations of both metals could overcome this decrease. Glutamine synthetase showed similar response under immobilized conditions compared to free cells with both metals. The addition of algal filtrate in 3:1 ratio further increased the nitrogenase activity compared with immobilized cells treated with sublethal doses of both metals. Immobilization facilitated higher uptake of nickel as compared to chromium. The observations of the present study clearly demonstrate the protective effect of immobilization on Aulosira fertilissima against Nickel and chromium toxicity. Rice field ecosystem thus possess a bidirectional natural metal ameliorating system where Aulosira mats act as a naturally immobilized system and the decay of Aulosira along with other cyanobacteria act as natural chelators protecting the rice plants from deleterious effects of the heavy metals. Most importantly is...

Physiological and biochemical alterations in Anabaena 7120 under iron stress.

Various physiological and biochemical process like growth, NO3- -uptake, nitrate reductase, glutamine synthetase and ATPases (Mg2+ and Ca2+ dependent) in the cyanobacterium Anabaena 7120 were observed under iron stress. Growth was found to be maximum in 50 microM Fe3+ added cells however, 20 microM Fe3+ (the Fe3+ concentration generally used for routine culturing of cyanobacterial cell in Chu 10 medium) incubation resulted in lower growth. Fe3+ starvation on the other hand showed very poor growth up to 4th day but once the growth started it reached at significant level on 7th day. Higher Fe3+ concentration reflected reduced growth with lethality at 500 microM Fe3+. Chlorophyll a fluorescence under Fe3+ stress reflected almost the similar results as in case of growth. However, the pigment was found to be more sensitive as compared to protein under Fe3+ stress. Similar results have been observed in case of NO3-uptake with only 80% reduction in nutrient uptake in 500 microM Fe3+ incubated cells. Nitrate reductase activity was lower in Fe3+ starved cells as compared to significant enzyme activity in 20 and 50 microM Fe3+ incubated cells. Similar to nitrate reductase, glutamine synthetase also showed maximum level in 50 microM Fe3+ added cells, however, higher Fe3+ concentration (300-500 microM ) resulted in reduced enzymatic activity. Glutamine synthetase activity was less sensitivity as compared to nitrate reductase activity under Fe3+ stress. ATPase (Mg2+ and Ca2+ dependent) always showed higher level with increasing Fe3+ concentration.

Metabolite and light regulation of metabolism in plants: lessons from the study of a single biochemical pathway

We are using molecular, biochemical, and genetic approaches to study the structural and regulatory genes controlling the assimilation of inorganic nitrogen into the amino acids glutamine, glutamate, aspartate and asparagine. These amino acids serve as the principal nitrogen-transport amino acids in most crop and higher plants including Arabidopsis thaliana. We have begun to investigate the regulatory mechanisms controlling nitrogen assimilation into these amino acids in plants using molecular and genetic approaches in Arabidopsis. The synthesis of the amide amino acids glutamine and asparagine is subject to tight regulation in response to environmental factors such as light and to metabolic factors such as sucrose and amino acids. For instance, light induces the expression of glutamine synthetase (GLN2) and represses expression of asparagine synthetase (ASN1) genes. This reciprocal regulation of GLN2 and ASN1 genes by light is reflected at the level of transcription and at the level of glutamine and asparagine biosynthesis. Moreover, we have shown that the regulation of these genes is also reciprocally controlled by both organic nitrogen and carbon metabolites. We have recently used a reverse genetic approach to study putative components of such metabolic sensing mechanisms in plants that may be conserved in evolution. These components include an Arabidopsis homolog for a glutamate receptor gene originally found in animal systems and a plant PII gene, which is a homolog of a component of the bacterial Ntr system. Based on our observations on the biology of both structural and regulatory genes of the nitrogen assimilatory pathway, we have developed a model for metabolic control of the genes involved in the nitrogen assimilatory pathway in plants

Nitrate levels and stages of growth in hypernodulating mutants of Lupinus Albus. II.Enzymatic activity and transport of N in the xylem SAP

The enzymatic study and transport of N in the xylem sap was carried out with a view to observing the influence of different nitrate levels and growth stages of the plant in chemically treated mutants of Lupinus albus. Several stresses induce a reduction in plant growth, resulting in the accumulation of free amino acids, amides or ureides, not only in the shoot, but also in the roots and nodules. Although enzyme activity is decisive in avoiding products that inhibit nitrogenase by ammonium, little is known about the mechanism by wich the xylem carries these products. However, this process may be the key to the function of avoiding the accumulation of amino acids in the cells of infected nodules. The behaviour of the enzymes nitrate reductase (NR), phosphoenolpyruvate carboxylase (PEPC), glutamine synthetase (GS) and nitrogen compounds derived from fixation, such as N-Ó-amino, N-ureides and N-amide in mutant genotypes were observed. The NR enzyme was highly influenced by the application of nitrate showing much higher values than those in the non-application of nitrate, independently of genotype, being that the NR, the best evaluation period was in the tenth week. The L-62 genotype characterized with nitrate-resistance, clearly showed that the enzyme PEPC is inhibited by presence of nitrate. The L-135 genotype (nor fix) showed GS activity extremely low, thus demonstrating that GS is an enzyme highly correlated with fixation. With regard to the best growth stage for GS, Lupinus albus should be evaluated in the seventh week.

Effect of nitrate on nodulation and n2-fixation of lupinus mutabilis mutants at different growth stages

Uma das principais técnicas usadas para estudar o efeito inibidor do NO3 na fixaçäo biológica do N2, tem sido o uso de mutantes de estirpes de Rhizobium ou de plantas hospedeiras que näo apresentam o padräo usual de nodulaçäo. Este estudo foi conduzido usando genótipos parentais e mutantes de Lupinus mutabilis, submetidos a dois níveis de NO3: 0 e 5mM, com duas épocas de colheita: sete e dez semanas após emergência. A concentraçäo de 5mM revelou-se da maior importância na inibiçäo da nodulaçäo inicial de Lupinus, apesar de os dados demonstrarem que os mutantes L-114 e L-105 foram mais eficientes quanto a nodulaçäo, N-total e atividade da nitrogenase que a cv. SCG-25, e podem permitir um aumento da fixaçäo de N2. Mas näo houve diferenças na atividade da glutamina sintetase (GS) dos nódulos entre os mutantes L-114, L-105 e a cv. SCG-25, quando as plantas näo receberam NO3, mas na presença de NO3 houve tendência para os mutantes, especialmente para os L-114, apresentarem maior atividade de GS. Isto indica que o mutante L-114 tem uma maior capacidade de assimilaçäo do N fixado, associada com um maior suprimento de carboidratos disponíveis, como indicado pela atividade da PEP-carboxilase. Portanto, a tolerância parcial ao NO3 apresentada pelos mutantes L-114 e L-105 está associada com o fenótipo hipernodulante. Esta sugestäo pode ser observada de reduçäo de acetileno, que foram maiores para o mutante L-114 que para a cv. SCG-25, na presença ou na ausência de NO3

A hyperoxidant state at the start of each developmental stage during Neurospora crassa conidiation

Neurospora crassa conidiation and other microorganism differentiation processes can be explained as a response to a hyperoxidant state. Many proteins, among them NADP-glutamate dehydrogenase and glutamine synthetase, were oxidatively modiried and degraded at the start of all three morphogenetic transitions of the conidiation process. A shift in the redox state of the main cellular electron carriers, from a highly reduced state to a predominantly oxidized one, occurred stepwise with a highly oxidizing event at the start of each transition. Catalase activity increased during cell differentiation in various microorganisms and one of two catalases accumulated to a high level in the differentiated cell. Catalases in Neurospora were oxidized in vitro and in vivo giving rise to active enzyme conformers. Carotenoids, which have antioxidant functions in fungi, were consumed in each morphogenetic stage and oxidized carotenoids accumulated in differentiated cells. Carotenoid biosynthesis was induced following oxidative stress and during conidiation in each new cell structure. Increased generation of reactive oxygen species was detected by chemiluminescence at the onset of all transitions during differentiation. Antioxidants inhibited both light emission and cell differentiation. These and other data from the literature are discussed in the context of the Dioxygen Avoidance Theory of cell differentiation.

Effect of lactose induced diarrhea on intestinal glutaminase and muscle glutamine synthetase activities in rats

O objetivo deste estudo foi avaliar o efeito da diarréia induzida por lactose na senzimas chaves do metabolismo de glutamina no músculo esqulético e no intestino delgado, em ratos. Comparados com os controles de peso pareado, os animais com diarréia apresentaram atividade maior de glutamina sintetase do músculo, concomitante com uma reduçäo na concentraçäo de glutamina nesse tecido, e uma queda na concentraçäo de glutamina arterial. Essas alteraçöes säo semelhantes àquelas relatadas por outros investigadores em condiçöes em que ocorre a proteólise muscular tais como, durante a fase pós-operatória e septicemia. Além dos dados que sugerem alteraçöes gerais no metabolismo da glutamina, um achado importante deste estudo foi a verificaçäo de aumento na atividade específica de glutaminase intestinal dependente de fosfato em ratos com diarréia. A alteraçäo da atividade dessa enzima näo tem sido demonstrada em diveresas condiçöes tais como, acidose, alcalose, aumento na ingestäo de gluamina através de água ou dieta, situaçöes que supostamente, poderiam interferir na sua atividade

Photorespiratory ammonia assimilation in the leaves of barley, sorghum and Moricandia arvensis.

Effects of methionine sulphoximine (MSX) on glutamine synthetase activity and ammonia accumulation in the leaves of barley, sorghum and Moricandia arvensis were studied. Glutamine synthetase activity of sorghum leaves was relatively resistant to MSX treatment during initial stages. All the three plant species exhibited considerable accumulation of ammonia arising from non-photorespiratory sources, in response to MSX treatment. Minimal estimates of the rate of glycine decarboxylation were made employing MSX and isonicotinyl hydrazide together in the treatment solution. Changes in the pool sizes of amino acids involved in photorespiratory nitrogen metabolism as affected by MSX have also been investigated.