Higher Glutamate Dehydrogenase Activity Research Articles

Polyamines Metabolism Interacts with γ-Aminobutyric Acid, Proline and Nitrogen Metabolisms to Affect Drought Tolerance of Creeping Bentgrass.

Due to increased global warming and climate change, drought has become a serious threat to horticultural crop cultivation and management. The purpose of this study was to investigate the effect of spermine (Spm) pretreatment on metabolic alterations of polyamine (PAs), γ-aminobutyric acid (GABA), proline (Pro), and nitrogen associated with drought tolerance in creeping bentgrass (Agrostis stolonifera). The results showed that drought tolerance of creeping bentgrass could be significantly improved by the Spm pretreatment, as demonstrated by the maintenance of less chlorophyll loss and higher photosynthesis, gas exchange, water use efficiency, and cell membrane stability. The Spm pretreatment further increased drought-induced accumulation of endogenous PAs, putrescine, spermidine, and Spm, and also enhanced PAs metabolism through improving arginine decarboxylases, ornithine decarboxylase, S-adenosylmethionine decarboxylase, and polyamine oxidase activities during drought stress. In addition, the Spm application not only significantly improved endogenous GABA content, glutamate content, activities of glutamate decarboxylase and α-ketoglutarase, but also alleviated decline in nitrite nitrogen content, nitrate reductase, glutamine synthetase, glutamate synthetase, and GABA aminotransferase activities under drought stress. The Spm-pretreated creeping bentgrass exhibited significantly lower ammonia nitrogen content and nitrite reductase activity as well as higher glutamate dehydrogenase activity than non-pretreated plants in response to drought stress. These results indicated beneficial roles of the Spm on regulating GABA and nitrogen metabolism contributing towards better maintenance of Tricarboxylic acid (TCA) cycle in creeping bentgrass. Interestingly, the Spm-enhanced Pro metabolism rather than more Pro accumulation could be the key regulatory mechanism for drought tolerance in creeping bentgrass. Current findings provide a comprehensive understanding of PAs interaction with other metabolic pathways to regulate drought tolerance in grass species.

Oxidation of Energy Substrates in Tissues of Fish: Metabolic Significance and Implications for Gene Expression and Carcinogenesis.

Fish are useful animal models for studying effects of nutrients and environmental factors on gene expression (including epigenetics), toxicology, and carcinogenesis. To optimize the response of the animals to substances of interest (including toxins and carcinogens), water pollution, or climate changes, it is imperative to understand their fundamental biochemical processes. One of these processes concerns energy metabolism for growth, development, and survival. We have recently shown that tissues of hybrid striped bass (HSB), zebrafish, and largemouth bass (LMB) use amino acids (AAs; such as glutamate, glutamine, aspartate, alanine, and leucine) as major energy sources. AAs contribute to about 80% of ATP production in the liver, proximal intestine, kidney, and skeletal muscle tissue of the fish. Thus, as for mammals (including humans), AAs are the primary metabolic fuels in the proximal intestine of fish. In contrast, glucose and fatty acids are only minor metabolic fuels in the fish. Fish tissues have high activities of glutamate dehydrogenase, glutamate-oxaloacetate transaminase, and glutamate-pyruvate transaminase, as well as high rates of glutamate uptake. In contrast, the activities of hexokinase, pyruvate dehydrogenase, and carnitine palmitoyltransferase 1 in all the tissues are relatively low. Furthermore, unlike mammals, the skeletal muscle (the largest tissue) of HSB and LMB has a limited uptake of long-chain fatty acids and barely oxidizes fatty acids. Our findings explain differences in the metabolic patterns of AAs, glucose, and lipids among various tissues in fish. These new findings have important implications for understanding metabolic significance of the tissue-specific oxidation of AAs (particularly glutamate and glutamine) in gene expression (including epigenetics), nutrition, andhealth, as well as carcinogenesis in fish, mammals (including humans), and other animals.

Effects of sodium selenite and L-selenomethionine on feed intake, clinically relevant blood parameters and selenium species in plasma, colostrum and milk from high-yielding sows

A field study in periparturient sows fed different dietary concentrations of either sodium selenite or L-selenomethionine (SeMet) was conducted to evaluate feed intake, haematological and biochemical parameters as well as to describe some key selenium (Se) species, namely selenoprotein P (SelP), selenoalbumin (SeAlb) and selenomethionine (SeMet) as well as total Se in plasma, colostrum and milk.Thirty-two sows were allotted to four treatments from 30 days (d) prepartum throughout on average a 32 d lactation period. Sodium selenite supplemented diets contained 0.40 and 0.60 mg Se/kg feed, while SeMet supplemented feed contained 0.26 and 0.43 mg Se/kg feed. Concentrations of sodium selenite and SeMet in complete feed exceeded the upper limits for total dietary Se and added organic Se, respectively, according to the European Union legislation. Blood samples were collected at initiation of the study, at farrowing and at weaning. Colostrum samples were collected at farrowing and milk samples at weaning. Se species were subjected to liquid chromatography, and total Se and Se species were determined using inductively coupled plasma mass spectrometry.The SeMet supplemented diets resulted in higher feed intake and in higher levels of total Se, SelP, SeAlb and SeMet in colostrum compared with sows fed sodium selenite. Similar results were obtained for levels of total Se and SeMet in milk at weaning. The higher dietary sodium selenite concentration in sows’ feed did not increase the Se transfer into colostrum or milk when compared with those receiving the lower level of sodium selenite. However, the increase in serum-Zn from initiation until farrowing, observed in sows fed SeMet as well as the higher glutamate dehydrogenase activity in sodium selenite supplemented sows in this period might indicate a higher requirement of antioxidant defence in sodium selenite-supplemented sows.To our knowledge, the present data on Se species in plasma, colostrum and milk of sows represent the most complete investigation of Se in sows conducted to date. A higher amount of the above-mentioned Se species in the colostrum of sows supplemented with SeMet might strengthen the piglets’ antioxidative system and passive immunity as well as improve their average daily weight gain. The higher feed intake in sows fed diets supplemented with SeMet is an interesting finding that warrants further investigation.

Chlorogenic acids inhibit glutamate dehydrogenase and decrease intracellular ATP levels in cultures of chick embryo retina cells

Chlorogenic acids (CGAs) are a group of phenolic compounds found in worldwide consumed beverages such as coffee and green tea. They are synthesized from an esterification reaction between cinnamic acids, including caffeic (CFA), ferulic and p-coumaric acids with quinic acid (QA), forming several mono- and di-esterified isomers. The most prevalent and studied compounds are 3-O-caffeoylquinic acid (3-CQA), 4-O-caffeoylquinic acid (4-CQA) and 5-O-caffeoylquinic acid (5-CQA), widely described as having antioxidant and cell protection effects. CGAs can also modulate glutamate release from microglia by a mechanism involving a decrease of reactive oxygen species (ROS). Increased energy metabolism is highly associated with enhancement of ROS production and cellular damage. Glutamate can also be used as an energy source by glutamate dehydrogenase (GDH) enzyme, providing α-ketoglutarate to the tricarboxylic acid (TCA) cycle for ATP synthesis. High GDH activity is associated with some disorders, such as schizophrenia and hyperinsulinemia/hyperammonemia syndrome. In line with this, our objective was to investigate the effect of CGAs on GDH activity. We show that CGAs and CFA inhibits GDH activity in dose-dependent manner, reaching complete inhibition at high concentration with IC50 of 52 μM for 3-CQA and 158.2 μM for CFA. Using live imaging confocal microscopy and microplate reader, we observed that 3-CQA and CFA can be transported into neuronal cells by an Na+-dependent mechanism. Moreover, neuronal cells treated with CGAs presented lower intracellular ATP levels. Overall, these data suggest that CGAs have therapeutic potential for treatment of disorders associated with high GDH activity.

Optimization of γ-polyglutamic acid synthesis using response surface methodology of a newly isolated glutamate dependent Bacillus velezensis Z3.

A new glutamate-dependent γ-polyglutamic acid (γ-PGA) producer Z3 isolated from soil samples in Daxinganling forest region of China was identified, and its optimal medium components were investigated using response surface methodology. Strain Z3 was identified as Bacillus velezensis by physiology and biochemistry and 16S rDNA sequence analysis. This is the first report of glutamate-dependent B. velezensis with the ability to synthesize γ-PGA. Then, the optimum γ-PGA yield (5.58g/L) was achieved with glutamate 86g/L, glucose 36g/L, yeast extract powder 5.5g/L, and NaH2PO4 7.5g/L. Furthermore, activities of enzymes participating in glutamate synthesis were assessed, and the results showed that lower ketoglutaric dehydrogenase activity (KGDH) and higher glutamate dehydrogenase activity (GDH) resulted in higher γ-PGA yield. Identification of glutamate-dependent γ-PGA producer named B. velezensis Z3 enriches microbiological resources with γ-PGA-producing capacity. B. velezensis optimization of nutrients and analysis of enzymes activities will not only help to increase γ-PGA productivity but also to understand the γ-PGA synthesis mechanism in B. velezensis Z3.

Formation of volatile compounds, peptidolysis and carbohydrate fermentation by mesophilic lactobacilli and streptoccocci cultures in a cheese extract

Many studies about the influence of Lactococcus lactis on cheese flavour have been reported, while the impact of mesophilic lactobacilli and Streptococcus thermophilus, either as single or mixed cultures, have been less studied. The ability of Lactobacillus paracasei 90, Lb. casei 72 (INLAIN collection) and S. thermophilus 2 (commercial strain) to produce flavour-related biochemical changes, as single or mixed (lactobacilli + streptococci) cultures, was assessed in a cheese model. These three strains, with different activities of glutamate dehydrogenase (GDH) and aminotransferases (AT), were incubated (14d/37 °C) alone or in mixed cultures in a cheese model consisting of a sterile extract of a fresh cheese. Mesophilic lactobacilli showed fermentation of carbohydrates, an increase of peptidolysis and production of volatile compounds that were correlated with their AT activities. In addition, these strains also produced completely different profiles of the amino acceptor compounds: pyruvate and α-ketoglutarate. S. thermophilus 2 increased the level of α-ketoglutarate due to its high GDH activity, but its contribution to flavour compound production was negligible; small additional changes existed when S. thermophilus 2 was mixed with the lactobacilli. Biochemical changes leading to flavour formation were mainly due to the activity of lactobacilli. Our results suggest that the AT profile and the peptidolytic activity of each strain influenced the volatilome of the extracts during incubation. So, both lactobacilli strains could be used as adjunct cultures in cheeses to increase/diversify the flavour, but more studies are needed to deepen the knowledge about the potential of S. thermophilus 2 for the production of flavour compounds.

Genetic Examination of Initial Amino Acid Oxidation and Glutamate Catabolism in the Hyperthermophilic Archaeon Thermococcus kodakarensis

Amino acid catabolism in Thermococcales is presumed to proceed via three steps: oxidative deamination of amino acids by glutamate dehydrogenase (GDH) or aminotransferases, oxidative decarboxylation by 2-oxoacid:ferredoxin oxidoreductases (KOR), and hydrolysis of acyl-coenzyme A (CoA) by ADP-forming acyl-CoA synthetases (ACS). Here, we performed a genetic examination of enzymes involved in Glu catabolism in Thermococcus kodakarensis. Examination of amino acid dehydrogenase activities in cell extracts of T. kodakarensis KUW1 (ΔpyrF ΔtrpE) revealed high NADP-dependent GDH activity, along with lower levels of NAD-dependent activity. NADP-dependent activities toward Gln/Ala/Val/Cys and an NAD-dependent threonine dehydrogenase activity were also detected. In KGDH1, a gene disruption strain of T. kodakarensis GDH (Tk-GDH), only threonine dehydrogenase activity was detected, indicating that all other activities were dependent on Tk-GDH. KGDH1 could not grow in a medium in which growth was dependent on amino acid catabolism, implying that Tk-GDH is the only enzyme that can discharge the electrons (to NADP(+)/NAD(+)) released from amino acids in their oxidation to 2-oxoacids. In a medium containing excess pyruvate, KGDH1 displayed normal growth, but higher degrees of amino acid catabolism were observed compared to those for KUW1, suggesting that Tk-GDH functions to suppress amino acid oxidation and plays an anabolic role under this condition. We further constructed disruption strains of 2-oxoglutarate:ferredoxin oxidoreductase and succinyl-CoA synthetase. The two strains displayed growth defects in both media compared to KUW1. Succinate generation was not observed in these strains, indicating that the two enzymes are solely responsible for Glu catabolism among the multiple KOR and ACS enzymes in T. kodakarensis.

Quantitative in silico Analysis of Neurotransmitter Pathways Under Steady State Conditions

The modeling of glutamate/GABA-glutamine cycling in the brain tissue involving astrocytes, glutamatergic and GABAergic neurons leads to a complex compartmentalized metabolic network that comprises neurotransmitter synthesis, shuttling, and degradation. Without advanced computational tools, it is difficult to quantitatively track possible scenarios and identify viable ones. In this article, we follow a sampling-based computational paradigm to analyze the biochemical network in a multi-compartment system modeling astrocytes, glutamatergic, and GABAergic neurons, and address some questions about the details of transmitter cycling, with particular emphasis on the ammonia shuttling between astrocytes and neurons, and the synthesis of transmitter GABA. More specifically, we consider the joint action of the alanine-lactate shuttle, the branched chain amino acid shuttle, and the glutamine-glutamate cycle, as well as the role of glutamate dehydrogenase (GDH) activity. When imposing a minimal amount of bound constraints on reaction and transport fluxes, a preferred stoichiometric steady state equilibrium requires an unrealistically high reductive GDH activity in neurons, indicating the need for additional bound constants which were included in subsequent computer simulations. The statistical flux balance analysis also suggests a stoichiometrically viable role for leucine transport as an alternative to glutamine for replenishing the glutamate pool in neurons.

In vitro and in situ activity of carboxymethyl cellulase and glutamate dehydrogenase according to supplementation with different nitrogenous compounds

Two experiments were carried out to evaluate the effect of supplementation with different nitrogenous compounds on the activities of carboxymethil cellulase (CMCase) and glutamate dehydrogenase (GDH). In the first experiment, four treatments were evaluated in vitro: cellulose, cellulose with casein, cellulose with urea, and cellulose with casamino acids. After 6, 12 and 24 hours of incubation, CMCase and GDH activity, pH, and concentrations of ammonia nitrogen (AN) and microbial protein were measured. In the three incubation periods, the concentration of AN was higher when urea was used as a supplemental source of nitrogen. The activity of CMCase was higher with the addition of urea and casamino acids when compared with the control and the casein treatment. Supplementation with casamino acids provided higher GDH activity when compared with the control at 6 hours of incubation. At 12 hours of incubation, the GHD activity was also stimulated by casein. At 24 hours, there was no difference in GHD activity among treatments. In the second experiment, three rumen-fistulated bulls were used for in situ evaluation. Animals were fed Tifton hay (Cynodon sp.) ad libitum. The treatments consisted of control (no supplementation), supplementation with non-protein nitrogenous compounds (urea and ammonium sulphate, 9:1) and supplementation with protein (albumin). In treatments with nitrogenous compound supplementation, 1 g of crude protein/kg of body weight was supplied. The experiment was conducted in a 3 × 3 Latin square design. The measurements were performed at 6, 12 and 24 hours after supplementation. No difference in GDH activity was observed among treatments. The control treatment showed higher CMCase activity when compared with the treatments containing supplemental sources of nitrogen. However, urea supplementation provided higher CMCase activity compared to albumin.

Liver-related biochemical changes in the serum of dogs being treated with phenobarbitone

The concentrations of phenobarbitone, albumin, bile acids and cholesterol, and the activities of alkaline phosphatase (AP), alanine aminotransferase (ALT), gamma glutamyl transferase (GGT) and glutamate dehydrogenase (GLDH) were measured in...

Lipid, Carbohydrates and Protein Patterns During Metamorphosis of the Mediterranean Fruit Fly, <I>Ceratitis capitata</I> (Diptera: Tephritidae)

We investigated the patterns of total lipid, glycogen, hemolymph carbohydrate, and trichloroacetic acid-precipitable protein contents starting 8 h before puparium formation and throughout the whole metamorphosis of the Medfly, Ceratitis capitata (Wiedemann). In addition, during the early stages of metamorphosis, we measured the activity of acid proteinases and glutamate dehydrogenase (GDH), an important enzymatic link between amino acids and the tricarboxylic acid cycle. Lipids accumulate during the larval-dispersal phase and the first 20 h of the prepupal stage. Thereafter, levels decline slightly throughout the pupal stage and first half of the pharate-adult stage, and drastically decline toward the last hours of metamorphosis. Glycogen and hemolymph carbohydrates are highly used from the moment the larvae leave the diet until the mid-pupal stage, then glycogen increases toward the mid pharate-adult and decreases again to low levels at the moment of adult emergence. Protein levels decrease during the prepupal stage (coinciding with a rise in acid proteinase activity) recovering toward the pharate-adult stage. GDH activity increases during the prepupal stage, attaining maximal activity at the onset of the pupal stage and then declines to low levels. This high GDH activity may be linked to the conversion of glutamate to α-ketoglutarate while proteins are being broken during tissue histolysis. Our results are discussed in view of the differential utilization of metabolites during metamorphosis.

Purification and characteristics of glutamate dehydrogenase (GDH) from triticale roots

In the investigated 14 day old triticale seedlings a much higher GDH activity was observed in roots than in leaves. The enzyme from the roots was purified up to the state of homogeneity (about 400 fold). The purified enzyme showed a higher activity in the presence of reduced coenzyme forms (NAD(P)H) than their oxidated forms. In the presence of NAD(P)H the enzyme showed absolute specificity to 2-oxoglutarate and in cooperation with NAD(P)+ to L-glutamate. The Km values determined for particular substrates indicate a high affinity of NADPH-GDH to ammonium ions.

Clinicopathological features of equine primary hepatic disease: a review of 50 cases

The clinicopathological features of 50 cases of equine hepatic disease were reviewed. There was a wide range of clinical signs and at least 50 per cent of the animals exhibited...

In situ measurement of glutamate concentrations in the periportal, intermediate, and pericentral zones of rat liver.

We developed a quantitative histochemical assay for measurement of local glutamate concentrations in cryostat sections of rat liver. Deamination of glutamate by glutamate dehydrogenase (GDH) was coupled to the production of formazan and formazan precipitation was used for colorimetric visualization. The method was tested and validated with gelatin model sections with known glutamate concentrations. Calibration graphs showed linear relationships with high correlation coefficients (> 96%) between glutamate concentrations or section thickness and absorbance values. The method was reproducible, with a constant percentage of 60 +/- 5% of glutamate being converted in gelatin model sections containing glutamate concentrations of 2 mM and higher. Glutamate concentrations were estimated in periportal, intermediate, and pericentral zones of liver lobules that contain low, intermediate, and high GDH activity, respectively. In fed adult male rat livers, periportal zones contained the highest concentrations of glutamate (approximately 14 mM) and intermediate and pericentral zones approximately 13 and 9 mM, respectively. On starvation, glutamate concentrations increased only in the small rim of pericentral cells that express glutamine synthetase, to approximately 15 mM. In livers of fetal and newborn rats, glutamate was homogeneously distributed, with a concentration of approximately 5 mM. In suckling rat liver, distribution of glutamate was still homogeneous but the concentration was increased to approximately 8 mM. These glutamate distribution patterns were in agreement with those detected immunohistochemically.

Susceptibility of primary cultures of proximal tubular and distal tubular cells from rat kidney to chemically induced toxicity

Isolated proximal tubular (PT) and distal tubular (DT) cells from rat kidney were cultured for up to 9 days under serum-free, hormonally-defmed conditions on 35-mm polystyrene culture dishes. Several hormonal and growth factor supplements were assessed for their ability to promote growth (increased protein and DNA content) and stability of differentiated phenotype (high activities of γ-glutamyltransferase and alkaline phosphatase as brush-border membrane markers in PT cells; maintenance of high activities of glutamate dehydrogenase as a mitochondrial marker in both PT and DT cells; maintenance of low and high activities of lactate dehydrogenase in PT and DT cells, respectively; expression of cytokeratins). Basal supplemented media (DMEM/F12, 1:1 v/v) contained insulin, hydrocortisone, epidermal growth factor, sodium selenite and transferrin as supplements. Additionally, triiodothyronine selectively promoted growth and stability of differentiated phenotype in PT cells and thyrocalcitonin selectively promoted growth and stability of differentiated phenotype in DT cells. On Day 3 of primary culture, PT and DT cells were incubated for up to 8 h with either ten-butyl hydroperoxide (tBH; 0.5–100 mM), methyl vinyl ketone (MVK; 1–10 mM), or p-aminophenol (PAP; 1−10 mM) and cellular injury, as assessed by cellular release of lactate dehydrogenase, was determined. DT cells were significantly more susceptible to injury from both tBH and MVK, but the two cell populations were equally susceptible to injury from PAP, which is the same susceptibility pattern seen in freshly isolated cells. These results suggest that primary cultures of rat renal PT and DT cells reflect similar biochemical properties as freshly isolated cells and are, therefore, useful models for study of chemically induced injury.

Disturbed nitrogen metabolism associated with the hyperhydric status of fully habituated callus of sugarbeet

The content of polyamines and proline was much lower in a normal (N) callus of Beta vulgaris L. than in a fully habituated hyperhydric (H) callus. The H callus also contained more glutamate and had a higher glutamate dehydrogenase activity. The excess of glutamate, in this chlorophyll‐deficient callus, was linked to accumulation of proline and polyamines. Experiments with α‐difluoromethylornithine (DFMO) and α‐difluoromethylarginine (DFMA) showed that both ornithine decarboxylase and arginine decarboxylase participated in the synthesis of polyamines (especially spermidine and putrescine) and removal of ammonia. It is hypothesized that the H callus was subjected to ammonia stress from the start of the culture. Experiments with gabaculine, an inhibitor of ornithine aminotransferase, showed that this enzyme linked proline degradation to polyamine synthesis through the production of ornithine. This disturbed nitrogen metabolism appeared to be characteristic of the fully habituated callus and might explain the low growth of this hyperhydric tissue.

Enzymes of ammonium assimilation inStreptomyces avermitilis

Glutamine synthetase (GS), glutamate synthase (GOGAT), glutamate dehydrogenase (GDH), alanine dehydrogenase (ADH) and alanine aminotransferase (GPT) were detected in the cell-free homogenate ofStreptomyces avermitilis grown in a defined medium containing ammonium sulfate as the only nitrogen source. At an initial NH4 + concentration of 7.5 mmol/L, high activities of GS, GOGAT and GDH were found while that of ADH was low. The ADH activity was markedly increased at initially millimolar NH4 + concentrations. In some characteristics of its NH4 +-assimilating system (e.g. control of some enzyme activities, the NADPH specificity of GOGAT, the presence of alanine aminotransferase),S. avermitilis differs from other known streptomycetes.

Comparison of properties of glutamate dehydrogenases in members of the Bacteroides fragilis group.

Dual coenzyme-specific glutamate dehydrogenase (GDH) is regulated by extracellular concentrations of ammonia in members of the Bacteroides fragilis group, including B. fragilis, B. ovatus, B. distasonis, and B. vulgatus, as shown previously in B. fragilis (YAMAMOTO et al., J. Gen. Appl. Microbiol., 30, 499 (1984); J. Gen. Microbiol., 133, 2773 (1987)). High specific activities of GDH were observed in extracts prepared from the respective species grown with limited amounts of ammonia, and GDHs were inactivated by adding NH4Cl at high concentrations to cultures growing with limited amounts of ammonia. The values of Km of GDHs from the bacterial species were in ranges of 1-5mM, 0.14-0.7mM, and 0.006-0.02mM for NH4Cl, 2-oxoglutarate, and NADPH, respectively, and the optimum pHs were around 8.0 in the reductive amination. GDH activities in the four species were inhibited to the same extent by an antiserum prepared against the B. fragilis GDH, and immunoprecipitin bands fused with each other without a spur in a double immunodiffusion test. Activities of glutamine synthetase and glutamate synthase were also found but were 1% of the NADPH-activities of GDH in all the bacterial species.

MASKING BY ENZYME INHIBITOR OF RAISED SERUM GLUTAMATE DEHYDROGENASE ACTIVITY IN REYE'S SYNDROME

Serum glutamate dehydrogenase (GDH) activity was greatly raised (up to 830 times the upper limit of normal) in 16 patients with Reye's syndrome. The serum activity was masked by an inhibitor, and the rises were observed only after dialysis or sample dilution. Serum GDH values from 38 paediatric patients, including 10 with hyperammonaemia due to other causes, showed no such rise after dialysis. Only 1 of 13 adult patients with liver disease had high GDH activity, but this level was not increased after dialysis. Serum ornithine carbamyl transferase activity was also raised in patients with Reye's syndrome, but levels were not increased after dialysis. The ratio of dialysed/undialysed GDH activity clearly distinguished all Reye's patients from controls. The inhibition of a mitochondrial enzyme which regulates ammonia metabolism may contribute to the hyperammonaemia of Reye's syndrome. Serum GDH levels before and after dialysis would seem to be a useful diagnostic aid in Reye's disease.

Changing Activities of Nitrogen-assimilating Enzymes During Growth and Senescence of Dwarf Beans (Phaseolus vulgaris L.)

Nitrate reductase, glutamine synthetase, glutamate synthase and glutamate dehydrogenase were measured in extracts of leaves, pods and seeds of Phaseolus vulgaris L. grown hydroponically with 16 mM nitrate as nitrogen source. Nitrate reductase activity per leaf was highest in young expanding and well illuminated leaves, and decreased rapidly in the lower leaves after full expansion. Low nitrate reductase activity was found in senescing leaves and pods. No nitrate reductase activity was detectable in seeds. Glutamine synthetase and methylviologen dependent glutamate synthase both increased in activity with leaf expansion, remained active longer than nitrate reductase and decreased during leaf nitrogen mobilization. Glutamine synthetase and glutamate synthase activities were low in pods, and in seeds, only glutamine synthetase was measurable with the methods used. Glutamate dehydrogenase was present in all organs analyzed. In leaves, glutamate dehydrogenase activity increased in fully expanded leaves and remained fully active during leaf senescence. Nitrate reductase activity was always lowest of the enzymes measured. The time courses of extractable nitrate reductase activity suggest that nitrate assimilation within the shoot was most active in developing and well illuminated leaves. Since glutamine synthetase and glutamate synthase remained active longer than nitrate reductase in the individual leaves, these two enzymes could continue to reassimilate ammonia released from mitochondria during photo-respiration. The high glutamate dehydrogenase activity in senescing leaves is consistent with its possible function in the oxidation of glutamate.