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PubMed1996

Coronary flow and flow reserve by PET simplified for clinical applications using rubidium-82 or nitrogen-13-ammonia.

UNLABELLED: To validate routine, noninvasive determination of absolute myocardial perfusion and coronary flow reserve (CFR), cardiac PET was performed in animals using a simplified imaging protocol, high-dose dipyridamole and a simplified quantitative algorithm specific for 82Rb and 13N-ammonia. METHODS: One hundred thirty-five PET scans were obtained in eight dogs after intravenous 13N-ammonia or 82Rb using serial dynamic PET or a simple two-image dataset. A simple flow model using the two-image dataset was developed for each radionuclide to account for varying arterial input function, flow-dependent myocardial extraction and increased permeability surface area (PS) product due to capillary recruitment at high flows not incorporated into previous models. Myocardial perfusion by the simple model was compared to standard, complete, two-compartment kinetic models validated by comparison to electromagnetic flow meter. RESULTS: For 13N-ammonia, myocardial perfusion by the simple PET model correlated with that by complete compartmental analysis of multiple serial PET images with r = 0.94, slope = 0.96; CFR by compartmental analysis correlated with CFR by electromagnetic flow meter with r = 0.94, slope = 0.97. For 82Rb, myocardial perfusion determined by the simple model correlated with that determined by complete compartmental analysis of multiple serial PET images with r = 0.98, slope = 1.06; CFR determined by compartmental analysis correlated with CFR by electromagnetic flow meter with r = 0.88, slope = 1.13. CONCLUSION: A simplified PET protocol using 13N-ammonia or 82Rb and simple flow models provide noninvasive measurement of CFR up to six times baseline flow throughout the heart and diagnostic image quality for routine clinical application.

Ammonia↗
PubMed1996

Effect of sodium bisulfate on ammonia concentration, fly population, and manure pH in a horse barn.

OBJECTIVE: To evaluate the effectiveness of altering the pH of manure with sodium bisulfate, thereby decreasing ammonia concentration and fly population in a horse barn environment. ANIMALS: 4 mixed-breed pony stallions. PROCEDURE: The 4-week study was scheduled with 2 weeks of treatment (with 2 application rates) and 2 weeks with no treatment (control weeks). During treatment weeks, sodium bisulfate was applied daily to the top of the bedding and straw, then spread on top. Ponies were kept in the stalls 24 hours a day during the 7-day test period and stall cleaning was not done. On day 7 of each week, ammonia concentration, manure pH, number of flies on a fly tape, and fly-evasive behavior patterns were determined. RESULTS: Sodium bisulfate applied to the horse stall environment daily of either 2.3 or 4.5 kg/9.3 m2 (5 or 10lb/ 100 ft2) decreased ammonia concentration, manure pH, and number of flies in the stall environment, compared with a control period with no sodium bisulfate. Fly-evasive behavior patterns of ponies occupying the stalls, including tail swishes, head tosses, and kicks/strikes, were decreased during the period of sodium bisulfate application. CONCLUSION: Sodium bisulfate may be an effective method of decreasing ammonia concentration and may serve as a method of fly control in horse barns.

Ammonia↗
PubMed1996

[Changes in the blood morphofunctional characteristics of cyprinid fishes in ammonia poisoning].

Changes in the carp blood and hemoglobin caused by action of threshold levels (0.1 mg/1 litre) of ammonia have been investigated. A decrease in the quantity of erythrocytes, increase in the amount of leucocytes and hematocrit are determined. Concentration of oxyhemoglobin falls. General hemoglobin, desoxyhemoglobin and, particularly, methemoglobin increase. Changes in the curve of dissociation of oxyhemoglobin with a shift to the right, which causes an increase in P50 are found. Ammonia induces an increase in hemoglobin sensibility to denaturation effects of acids and alkalis. Modification of functions and structure of hemoglobin is in close connection with such indices of blood and hemoglobin as pH, concentration of 2,3-DFG, ammonia, ATP, amid groups of proteins and proteins glycation. A conclusion is made that ammonia breaks homeostatic equilibrium of blood, its composition and structural functional properties of hemoglobin.

Ammonia↗
PubMedDate not supplied

Anaerobic ammonia oxidation with nitrogen dioxide by Nitrosomonas eutropha.

Nitrosomonas eutropha, an obligately lithoautotrophic bacterium, was able to nitrify and denitrify simultaneously under anoxic conditions when gaseous nitrogen dioxide (NO2) was supplemented to the atmosphere. In the presence of gaseous NO2, ammonia was oxidized, nitrite and nitric oxide (NO) were formed, and hydroxylamine occurred as an intermediate. Between 40 and 60% of the produced nitrite was denitrified to dinitrogen (N2). Nitrous oxide (N2O) was shown to be an intermediate of denitrification. Under an N2 atmosphere supplemented with 25 ppm NO2 and 300 ppm CO2, the amount of cell protein increased by 0.87 mg protein per mmol ammonia oxidized, and the cell number of N. eutropha increased by 5.8 x 10(9) cells per mmol ammonia oxidized. In addition, the ATP and NADH content increased by 4.3 micromol ATP (g protein)-1 and 6.3 micromol NADH (g protein)-1 and was about the same in both anaerobically and aerobically grown cells. Without NO2, the ATP content decreased by 0.7 micromol (g protein)-1, and the NADH content decreased by 1.2 micromol (g protein)-1. NO was shown to inhibit anaerobic ammonia oxidation.

Adenosine Triphosphate↗
PubMed1997

Effects of simulated upper gastrointestinal hemorrhage on ammonia and related amino acids in blood and brain of chronic portacaval-shunted rats.

Gastrointestinal (GI) hemorrhage during compromised liver function is known to precipitate portal-systemic encephalopathy (PSE). Hypothetically, the induced hyperammonemia depletes cerebral glutamate pools. To investigate this hypothesis, rats were studied 14 days after portacaval shunt (PCS) or sham surgery (SHAM). Rats received 3 mL bovine erythrocytes or saline at t = 0, 1, 2, and 3h via a previously placed gastrostomy catheter. At t = 0, 2, 4, 6 and 8h arterial blood and at t = 8h cerebral cortex were sampled for determination of ammonia and amino acids. Control rats (NORM) were sampled without previous surgery. Repeated intragastric blood administration increased the already elevated arterial ammonia levels in PCS rats further. This resulted in higher cerebral cortex ammonia and glutamine levels after blood administration. Despite the accumulation of ammonia and glutamine, cerebral cortex glutamate concentrations remained unaltered. Yet, PCS rats became more encephalopathic after blood gavages, suggesting that there is not a clear-cut relation between cerebral cortex glutamate concentrations and degree of PSE. Interestingly, cerebral cortex concentrations of GABA, tyrosine and phenylalanine were markedly increased. Whether these observations are pathogenetically related to PSE remains to be established. The present model of simulated GI hemorrhage in PCS rats seems to be a suitable, clinically valid model for future research regarding hepatic encephalopathy.

Amino Acids↗
PubMed1997

Treatment of cirrhotic rats with L-ornithine-L-aspartate enhances urea synthesis and lowers serum ammonia levels.

CCl4-induced cirrhosis of rats was used for studying the influence of L-ornithine-L-aspartate (OA) on hyperammonemia. OA given to cirrhotic rats (2 g/kg daily) for 2 wk slightly increased net body weight and led to a significant increase in plasma urea levels and a decrease in plasma ammonia levels. Serum concentrations of glutamate, glutamine and arginine decreased significantly. In the livers of the OA-treated rats the activities of carbamoylphosphate synthetase I and arginase increased by 30 and 40%, respectively, approaching normal levels. No change in the activities of the other urea cycle enzymes as well as of glutamate dehydrogenase, glutaminase and glutamine synthetase was found. The negative correlation between glutamine synthetase activity and plasma ammonia levels reported previously for cirrhotic rats (Gebhardt and Reichen, Hepatology 20:684-691, 1994) was corroborated for cirrhotic animals not treated with OA, but was no longer apparent in OA-treated cirrhotic rats. Despite this improvement, plasma ammonia levels still varied considerably reflecting the variable accessibility and activities of glutamine synthetase in cirrhotics. Cultured hepatocytes from the two groups of rats showed a similar stimulation of urea production by addition of ammoniumacetate and/or OA to Hanks' buffered salt solution. In Williams medium E, however, the hepatocytes from the OA group produced significantly more urea than those from controls. These results suggest that treatment of cirrhotic rats with OA considerably improves urea production favoring the detoxification of ammonia that, however, is still limited by the severe alterations in liver architecture that are not influenced by OA in a 2-wk period.

Amino Acids↗
PubMedDate not supplied

[Effect of hypothermia and hyperoxia on the ammonia-glutamic acid system in the brain of rats].

The content of ammonia, glutamine, dicarboxylic amino acids and GABA was studied in the brain under 1, 2, 4-fold separate and simultaneous effect of hypothermia (19-20 C) and hyperoxia (3 atm.). A two-fold hypothermia of rats is accompanied by a greater increase of ammonia in the brain than a three-fold one. The content of glutamine under two-fold cooling is unchanged and under three-fold cooling it is twice as low as compared to its content in the brain of the control rats. The content of glutamic acid decreased after two-fold hypothermia is almost unchanged by the third seance of hypothermia. The repeated actions of hyperoxia also cause a considerable increase in the ammonia content but the dynamics of changes in the content of the nitrogenous metabolic products is contary to that in animals subjected to repeated seances of hypothermia. A simultaneous combined action of hypothermia and hyperoxia produces no additive effect on the system ammonia-glutaminic acid.

Ammonia↗
PubMedDate not supplied

Rapid determination of volatile bases in fish by using an ammonia ion-selective electrode.

A simple and rapid method using an ammonia ion-selective electrode (ISE) to measure volatile bases in fish is proposed. Accuracy and precision were determined with 5, 10, 20, and 30 ppm NH3 standard solutions. Ammonia values obtained with the method correlate strongly with total volatile basic nitrogen (r2 = 0.88). Recoveries of added ammonia to homogenized fish samples ranged from 83.7 to 96.0%. Responses of the probe to trimethylamine (TMA), calculated as NH3 (mg/100 mL), ranged from 74.9 to 91.7%. These findings indicate that the probe measured TMA as well as ammonia. Storage trials on 8 fish species illustrated that the results obtained with the ISE method reflected nitrogen concentrations based on total volatile base (TVB) analysis. This procedure may be used in lieu of the traditional TVB method for on-site rapid screening of fish.

Ammonia↗
PubMed2005

Friedel-Crafts-type mechanism for the enzymatic elimination of ammonia from histidine and phenylalanine.

The surprisingly high catalytic activity and selectivity of enzymes stem from their ability to both accelerate the target reaction and suppress competitive reaction pathways that may even be dominant in the absence of enzymes. For example, histidine and phenylalanine ammonia-lyases (HAL and PAL) trigger the abstraction of the nonacidic beta protons of these amino acids while leaving the much more acidic ammonium hydrogen atoms untouched. Both ammonia-lyases have a catalytically important electrophilic group, which was believed to be dehydroalanine for 30 years but has now been revealed by X-ray crystallography and UV spectroscopy to be a highly electrophilic 5-methylene-3,5-dihydroimidazol-4-one (MIO) group. Experiments suggest that the reaction is initiated by the electrophilic attack of MIO on the aromatic ring of the substrate. This incomplete Friedel-Crafts-type reaction leads to the activation of a beta proton and its stereospecific abstraction, followed by the elimination of ammonia and regeneration of the MIO group. The plausibility of such a mechanism is supported by a synthetic model. The application of the PAL reaction in the biocatalytic synthesis of enantiomerically pure alpha-amino beta-aryl propionates from aryl acrylates is also discussed.

Ammonia↗
PubMed2003

X-ray diffraction studies on mesophases of cetyl- and dodecyltrimethylammoniumbromide in liquid ammonia.

We have studied solutions of the surfactants cetyltrimethylammoniumbromide (CTAB) and dodecyltrimethylammoniumbromide (DTAB) in liquid ammonia with respect to the formation of lyotropic phases. For this purpose, a set-up for performing X-ray scattering experiments at temperatures up to 120 degrees C on samples containing liquid ammonia has been developed. Both systems form hexagonal and monoclinic lyotropic phases above the dissolving temperature of the surfactant, thus representing the first examples for lyotropic phases in liquid ammonia, and for monoclinic phases in nonaqueous solvents. The phase diagrams of CTAB/liquid NH(3) and DTAB/liquid NH(3) show similarities to their respective aqueous systems. However, the regions of existence of monoclinic phases are much larger in the ammonia system, while the cubic phases, as observed in the water based systems, do not seem to exist. The liquid-crystalline phases found provide potentiality for preparing mesoporous, nitride-based solids.

Journal Article↗
PubMed2001

Reactivity of anomeric 1alpha- and 1beta-pentofuranosyl azide derivatives in ammonia chemical ionization.

Electron impact (EI), fast atom bombardment (FAB) and ammonia chemical ionization [CI(NH3)] mass spectrometry were applied with the aim of differentiating between the anomeric 1alpha- and 1beta-azidopentofuranosyl derivatives. Calculated ammonium affinities [AA(M)] and proton affinities [PA(M)] show that beta-anomers have higher affinities for H+ and NH4+ ions than alpha-azides. Protonated molecules, obtained by CI(NH3) of azidofuranosyl derivatives, lose HN3 giving abundant furanosyl (S+) ions. Ammonia solvation of MH+ ions competes with the previous reaction producing the [SNHN2NH3]+ ion, a competitive product to the ammonium-attached [SN3NH4]+ ion. The fragmentation pathways of the stable and metastable [MNH4]+, MH+ ions, and several other important fragment ions, were determined using mass analyzed ion kinetic energy spectrometry (MIKES). The abundance of the [SN3NH4]+ and/or [SNHN2NH3]+ ions was found to correlate inversely with the exothermicity of ammonia solvation of the MH+ ion. The abundance of the fragment ions [SNHNH3]+, [SNH3]+ and SNH+ in some examples correlates with the exothermicity of the corresponding [MNH4]+ and MH+ parent ion formation. The fragment ions SNH3+ and SNHNH3+ can be formed, at least in part, in the ammonia solvation reaction of the S+ and SNH+ ions taking place within the high-pressure region of the CI ion source.

Journal Article↗
PubMed1991

Ammonia-induced astrocyte swelling in primary culture.

The effect of ammonia on water space of astrocytes in culture was determined as a means of studying the neurotoxicity of ammonia in fulminant hepatic failure (FHF). Treatment of primary astrocyte cultures obtained from neonatal rat cortices with 10 mM NH4Cl for 4 days resulted in a 29% increase in astrocytic water space, as measured by an isotopic method utilizing 3-O-methyl-[3H]-glucose. This effect was time- and dose-dependent. The ammonia-induced swelling was reversible as the water space in cultures treated with 10 mH NH4Cl for 3 days, and then returned to normal culture media for 1 day, was similar to control cultures. These findings suggest that elevated levels of ammonia lead to astrocyte swelling and may contribute to the brain edema in FHF.

3-O-Methylglucose↗
PubMed1977

Regulation of ammonia uptake in aspergillus nidulans.

The ammonia uptake in A. nidulans was found to be linear for about 20 min, and was proportional up to 1.5 mg/ml dry cell density. The transport of ammonia does not involve energy. Normal and biotin deficient A. nidulans showed an identical Km-values of 10.26 X 10(-5) M ammonia for uptake. The uptake of ammonium ion has been shown to be regulated by the intracellular concentration of ammonia.

Aspergillus nidulans↗
PubMed1992

Growth study of lactate and ammonia double-resistant clones of HL-60 cells.

The possibility that lactate and ammonia accumulation may have less detrimental effect on cell growth than usually admitted is investigated. We report here the isolation of several HL-60 subclones able to proliferate in the presence of 60 mM sodium lactate and 4 mM ammonium chloride, concentrations usually considered to be toxic for cell proliferation. Growth kinetics and final cell densities of these clones in suspension cultures were similar to the HL-60 cell population in control medium as well as in medium containing ammonia and lactate in which control cells were unable to grow. The metabolic pattern of the double-resistant clones revealed that lactate and ammonia formation was inhibited in the presence of lactate and ammonia in the medium, while alanine production and arginine consumption were enhanced irrespective of the medium.

Ammonium Chloride↗
PubMed2001

Optical rubbery ormosils sensor for the detection of ammonia.

Rubbery ormosil films with immobilized aminofluorescein (AF) were investigated to develop an optochemical sensor for the determination of ammonia in water. The gel precursors with tetramethoxysilane (TMOS) and dimethyldimethoxysilane (DiMeDMOS) were deposited on glass supports, and characterized in terms of response to pH, and to dissolved ammonia at constant pH. After preconditioning the sensing film was stable for 6 months. The detection limit for ammonia in water was 0.2 microg mL(-1) (S/N 2), the response being linearly dependent on concentration in the range of 0.5 to 80 microg mL(-1) ammonia. The response time was less than 5 min. The effects of sodium chloride concentration, temperature, and coexisting metal ions and compounds were investigated.

Journal Article↗
PubMed1998

Functional Gene Hybridization Patterns of Terrestrial Ammonia-Oxidizing Bacteria.

Abstract The biochemical pathway and genetics of autotrophic ammonia oxidation have been studied almost exclusively in Nitrosomonas europaea. Terrestrial autotrophic ammonia-oxidizing bacteria (AAOs), however, comprise two distinct phylogenetic groups in the beta-Proteobacteria, the Nitrosomonas and Nitrosospira groups. Hybridization patterns were used to assess the potential of functional probes in non-PCR-based molecular analysis of natural AAO populations and their activity. The objective of this study was to obtain an overview of functional gene homologies by hybridizing probes derived from N. europaea gene sequences ranging in size from 0.45 to 4.5 kb, and labeled with 32P to Southern blots containing genomic DNA from four Nitrosospira representatives. Probes were specific for genes encoding ammonia monooxygenase (amoA and amoB), hydroxylamine oxidoreductase (hao), and cytochrome c-554 (hcy). These probes produced hybridization signals, at low stringency (30 degreesC), with DNA from each of the four representatives; signals at higher stringency (42 degreesC) were greatly reduced or absent. The hybridization signals at low stringency ranged from 20 to 76% of the total signal obtained with N. europaea DNA. These results indicate that all four functional genes in the ammonia oxidation pathway have diverged between the Nitrosomonas and Nitrosospira groups. The hao probe produced the most consistent hybridization intensities among the Nitrosospira representatives, suggesting that hao sequences would provide the best probes for non-PCR-based molecular analysis of terrestrial AAOs. Since N. europaea can also denitrify, an additional objective was to hybridize genomic DNA from AAOs with probes for Pseudomonas genes involved in denitrification. These probes were specific for genes encoding heme-type dissimilatory nitrite reductase (dNir), Cu-type dNir, and nitrous oxide reductase (nosz). No hybridization signals were observed from probes for the heme-type dNir or nosz, but Nitrosospira sp. NpAV and Nitrosolobus sp. 24-C hybridized, under low-stringency conditions, with the Cu-type dNir probe. These results indicate that AAOs may also differ in their mechanisms and capacities for denitrification.

Journal Article↗
PubMed1993

Assessment of the effects of dobutamine on myocardial blood flow and oxidative metabolism in normal human subjects using nitrogen-13 ammonia and carbon-11 acetate.

The dual purposes of this study with positron emission tomography were to measure the effects of dobutamine on myocardial blood flow and oxidative metabolism, and to compare carbon-11 (C-11) acetate versus nitrogen-13 (N-13) ammonia in quantitating flow in normal subjects. Flow was quantitated with N-13 ammonia at rest and at peak dobutamine infusion (40 micrograms/kg/min) in 21 subjects. In 11 subjects, oxidative metabolism was also estimated at rest and peak dobutamine infusion using the clearance rate of C-11 acetate, k mono (min-1). A 2-compartment kinetic model was applied to the early phase of the C-11 acetate data to estimate flow. The rest and peak dobutamine rate-pressure products were 7,318 +/- 1,102 and 19,937 +/- 3,964 beats/min/mm Hg, respectively, and correlated well (r = 0.77) with rest and peak dobutamine flows of 0.77 +/- 0.14 and 2.25 ml/min/g determined using N-13 ammonia as a flow tracer. Rest and dobutamine flows estimated with C-11 acetate were highly correlated with those determined with N-13 ammonia (r = 0.92). k mono increased from 0.05 +/- 0.01 to 0.18 +/- 0.02 min-1, and correlated highly with the increase in flows (r = 0.91) and rate-pressure products (r = 0.94). Thus, the increase in cardiac demand associated with dobutamine is highly correlated with an increase in supply and oxidative metabolism. C-11 acetate is a unique tracer that can be used to image both flow and metabolism simultaneously.

Adult↗
PubMed1990

Effect of ammonia on calcium homeostasis in primary astrocyte cultures.

Calcium influx, accumulation and efflux were studied in primary cultures of rat astrocytes treated with ammonium chloride. Treatment of the cells for 3 days with 10 mMN4Cl resulted in a 35% reduction in 45Ca influx. The decrease in calcium influx was dose-dependent between 2 and 10 mM NH4Cl. Short-term (30 min) exposure to ammonia had no effect on calcium influx. Calcium accumulation, as measured by 20-min exposure to 45Ca, decreased after treating cultures with 10 mM NH4Cl for one or 3 days; a greater effect was observed after the 3-day treatment. Studies with lanthanum, an inhibitor of calcium transport, indicated that the effect of ammonia was not due to non-specific leakage of calcium. Calcium efflux was not affected by exposure of the cultures to ammonium chloride. Purinergic-evoked calcium influx and mobilization was not altered by ammonia. While the mechanism(s) of calcium homeostasis affected by long-term hyperammonemia remain to be defined, these results suggest that reduced astrocytic calcium may be related to the pathogenesis of ammonia-related disorders such as hepatic encephalopathy.

Adenosine Triphosphate↗