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Ammonia monitoring at trace level using photoacoustic spectroscopy in industrial and environmental applications.

An ammonia traces analyser based on photoacoustic spectroscopy is described. The system uses a CO(2) laser and a properly designed resonant photoacoustic cell to achieve ammonia detection at sub-parts-per-billion (ppb) level. The instrument features unattended automatic on-line monitoring of ammonia with a detection limit of 0.1 ppb. Interferences from atmospheric CO(2) and H(2)O are efficiently suppressed by a careful selection of the laser wavelength and a compensation of the water vapour signal made with a high-precision hygrometer. The cell design enables continuous measurement at high flow rates (up to 5 l/min), which guarantees a fast response time of the system for the monitoring of ammonia, a sticky polar molecule that adheres to most surfaces. Various examples of applications of the instrument in the semiconductor industry and for atmospheric pollution monitoring are presented. They demonstrate the excellent performances of the system and its suitability for these applications.

Air↗

Biodegradation of natural and synthetic estrogens by nitrifying activated sludge and ammonia-oxidizing bacterium Nitrosomonas europaea.

This report describes the uses of nitrifying activated sludge (NAS) and ammonia-oxidizing bacterium Nitrosomonas europaea to significantly degrade estrone (E1), 17beta-estradiol (E2), estriol (E3), and 17alpha-ethynylestradiol (EE2). Using NAS, the degradation of estrogens obeyed first-order reaction kinetics with degradation rate constants of 0.056 h(-1) for E1, 1.3 h(-1) for E2, 0.030 h(-1) for E3, and 0.035 h(-1) for EE2, indicating that E2 was most easily degraded. Then, we confirmed that E2 was degraded via E1 by NAS. With/without the ammonia oxidation inhibitor, it was observed that ammonia-oxidizing bacteria in conjunction with other microorganisms in NAS degraded estrogens. Using N. europaea, the degradation of estrogens reasonably obeyed zero-order reaction kinetics, and no remarkable difference is present among the four estrogens degradation rates and it was found that E1 was not detected during E2 degradation period. We suggested that E2 was degraded to E1 in NAS could be caused by other heterotrophic bacteria, not by ammonia-oxidizing bacteria.

Ammonia↗

Lethal and sublethal toxicity of ammonia to native, invasive, and parasitised freshwater amphipods.

In lethal and sublethal ammonia toxicity tests, we examined differences in tolerance of three species of freshwater amphipods, one native and two invasive in Ireland. The native Gammarus duebeni celticus was slightly less tolerant to ammonia than the invasive G. pulex (96 h LC50= 1.155 and 1.544 mg l(-1), respectively), while another invader, Crangonyx pseudogracilis, had the lowest tolerance (LC50= 0.36 mg l(-1)). Parasitism of G. pulex by the acanthocephalan Echinorhynchus truttae greatly reduced the tolerance of the invader to ammonia (LC50= 0.381 mg l(-1)). Further, precopula pair disruption tests indicated that G. d. celticus was more sensitive to ammonia than G. pulex at sublethal levels. We discuss these results in the context of the ecological replacements of native by invader amphipods.

Adaptation, Physiological↗

Influence of operating conditions on the ammonia electro-oxidation rate in wastewaters from power plants (ELONITA technique).

Power plants, like many other industries, are confronted with the need to reduce the nitrogen content of their wastewaters. Indirect electro-oxidation, through hypochlorite formation, is a possible answer to an ammonia-nitrogen problem, through converting it into gaseous nitrogen. Several parameters affect the ammonia oxidation rate: current density, chloride concentration and the presence of oxygen-containing anions, mainly SO(4)(2-), CO(3)(2-) and PO(4)(3-). pH values between 5.5 and 10 have been found to have no effect on the ammonia oxidation rate. However, at higher pH values (above 11), the oxidation slows down and chlorate ions appear. A model can be fitted from the experiments to predict the ammonia oxidation rate based on four main parameters: pH, current density, sulfate concentration and chloride concentration. The average difference between the predicted oxidation rates and the experimental measures is only 6.5%. This model confirms that the optimal operating conditions are a high chloride concentration (7 gl(-1)), no sulfate and a high current density (1200 Am(-2)).

Ammonia↗

Effects of ammonia on synaptosomal membranes.

Acute and sustained hyperammonemia in mice resulted in a decrease of the transition temperature of Arrhenium plots of synaptosomal (Na+-K+)ATPase. The activation energies in both phases of the plots were increased. "In vitro" addition of ammonia produced similar changes. This seems to indicate that ammonia alters the physical properties of synaptosomal membranes. The "in vitro" interaction of ammonia and ethanol at the membrane level was also investigated. Both agents together produced a further shift in the transition temperature and affected the activation energies. The relevance of these findings regarding the mechanism of ammonia toxicity and the protective effect of ethanol thereon is discussed.

Acetates↗

Multilayered film analysis: evaluation of ammonia and creatinine slides.

We evaluated the Kodak Ektachem multilayer ammonia and creatinine slides. For various concentrations of control material the precision (CV) within- and (between-day) ranged from 0.8 to 7.8% (4.4-9.0%) for ammonia, and 1.9-25.1% (3.6-13.9%) for creatinine. A number of commercial control fluids were unacceptable choices due to the presence of elevated endogenous ammonia levels. For both analytes, analytical recovery was in the range of 100 to 108%. Comparison with the Jaffe reaction demonstrated good correlation for creatinine, and linearity was judged acceptable for both analytes. For ammonia, serum values were nearly double those of paired plasma samples. Plasma and serum samples, therefore, cannot be used interchangeably. No serum versus plasma differences were observed for creatinine.

Ammonia↗

Plasma ammonia levels in very low birth weight preterm infants.

The mean plasma ammonia level at birth of 36 very low birth weight infants (< or = 32 weeks of gestation) was 71 +/- 26 mumol/L (121 +/- 45 micrograms/dl), which is similar to the mean level in preterm infants born at > or = 32 weeks of gestational age. Plasma ammonia levels declined to 42 +/- 14 mumol/L (72 +/- 24 micrograms/dl) at 7 days of age; mean ammonia levels at 14, 21, and 28 days of age were similar to that at 7 days of age and to the mean plasma ammonia level of 14 healthy term infants at birth (45 +/- 9 mumol/L (77 +/- 16 micrograms/dl)).

Ammonia↗

Nitrification of high strength ammonia wastewaters: comparative study of immobilisation media.

Due to legislative pressures, sludge production and processing in the UK will increase substantially in the future resulting in a supernatant liquid high in ammonia (500-1000 mg l-1) and "hard" COD (approximately 500 mg l-1). A small footprint reactor is required to effectively nitrify this effluent, and the aim of this work was to compare a number of immobilisation media under a variety of conditions in order to determine which media held the most promise for future development. Laboratory-scale continuously stirred tank reactors containing freely suspended and immobilised biomass were operated with a high-strength synthetic ammonia wastewater (500 mg N l-1) to determine the nitrification rates at various temperatures, and ammonia and COD loadings. COD:NH3 ratios in sludge liquors vary widely depending on the treatment processes employed, and therefore ratios of 1:1 and 2:1 were tested as being fairly typical. The freely suspended nitrifiers were washed out of the reactors at a 1 d hydraulic retention time (HRT), whereas the reactors containing adsorption particles (Linpor and Kaldnes) and PVA-encapsulated nitrifiers continued partially nitrifying down to 12 h, and oxygen addition enhanced nitrification. A decrease in temperature from 25 to 16 degrees C only caused a small (10%) decrease in nitrification in the immobilised cell reactors, demonstrating that nitrification was mass transfer rather than kinetically controlled. A reduction in nitrification occurred when glucose (500 mg l-1) was added to the feed due to the growth of a heterotrophic population. The adsorbed biomass reactors lost 35% of nitrification compared to only 7% with PVA, and it appears that the colonisation of PVA by heterotrophs is more difficult than for Linpor and Kaldnes. Respiration rates for all particles increased with time in the reactors, and nitrifiers immobilised in PVA retained approximately 40% of their viability after immobilisation. Volumetric nitrification rates were generally higher for the PVA reactor than for Linpor and Kaldnes, and were: suspended biomass reactor: 0.36; Linpor: 0.57; Kaldnes: 0.53 and PVA: 0.70 kg N m-3-reactor d-1 for a 25% reactor fill. These equate to 2.28, 4.24 and 3.97 g N m-2-media d-1 for Linpor, Kaldnes and PVA respectively, hence other reactor fill rates for Kaldnes warrant further investigation. However, the PVA particles with the highest nitrification rates under all conditions showed promise as an immobilisation medium, and are amenable to further optimisation for the nitrification of high-strength ammonia wastewaters.

Ammonia↗

The effect of residual ammonia concentration under aerobic conditions on the growth of Microthrix parvicella in biological nutrient removal plants.

It is demonstrated with two parallel single reactor intermittently aerated nitrification denitrification systems fed municipal wastewater as influent, that Microthrix parvicella bulking can be stimulated and cured by manipulating the ammonia concentration in the aerobic period (by inhibiting the nitrifiers) to high and low values respectively. The proliferation or not of M. parvicella is hypothesized to be due to their requirement for ammonia as a nitrogen source for growth. In terms of this hypothesis, if nitrification is rapid and complete, ammonia is not freely available and will limit M. parvicella growth. If nitrification is not complete for whatever reason, ammonia is available for the growth of the slow growing M. parvicella, enabling their proliferation to cause a bulking sludge. This hypothesis does not overturn or replace the anoxic-aerobic (AA, or low Food/Microorganism, F/M, ratio) filament bulking hypothesis of Casey et al. (Water SA 25(4) (1999) 425) but appears to be additional to it. Future research will focus on determining how elements of both hypotheses superimpose on the conditions in BNR systems, to produce an AA filament bulking sludge or not.

Actinobacteria↗

Excretion of ammonia by Lucilia cuprina larvae suppresses immunity in sheep.

Attempts to immunise sheep against natural infestations by Lucilia cuprina larvae have not been effective. Yet it is known that the larvae excrete the immunosuppressant ammonium bicarbonate. The effect of larval ammonium and nonionic ammonia on immunopathobiology was evaluated in 12 infested sheep. The concentration of ammonium in veins draining infested sites was measured in another group of four sheep. Mean jugular unionized ammonia concentration increased 3.5 to 5.6 times above pre-infested control levels. Mean venous ammonium concentrations draining infested sites were 13 times higher than pre-infested jugular or carotid levels. Increases in jugular ammonia concentrations correlated with increased number of larvae, area of infestation, earlier death, neutropenia, eosinopenia, lymphocytopenia, large declines in serum globulins and zinc, and large rises in toxic neutrophils. The high concentrations of toxic unionized ammonia in blood directly permanently damaged neutrophils and lymphocytes and depressed serum globulin production. The results show that the ammonium from the excreta of larvae of L. cuprina may be highly immunosuppressive.

Ammonia↗

Kinetic characterization of the inactivation of ammonia monooxygenase in Nitrosomonas europaea by alkyne, aniline and cyclopropane derivatives.

The kinetic mechanisms of seven inactivators of ammonia oxidation activity in cells of the nitrifying bacterium, Nitrosomonas europaea were investigated. The effects of the inactivators were specific for ammonia monooxygenase (AMO) which oxidizes ammonia to hydroxylamine. The aniline derivatives, 1,3-phenylenediamine and p-anisidine, were potent inactivators of AMO while other derivatives were ineffective as inactivators. Two cyclopropane derivatives, 1, 2-dimethylcyclopropane and cyclopropyl bromide, were inactivators while cyclopropane was not an inactivator. The mechanisms of three alkynes, 1-hexyne, 3-hexyne, and acetylene, were also examined. For all seven compounds, the inactivation of AMO was irreversible, time-dependent, first-order, and dependent on catalytic turnover. Saturation of the rate of inactivation was indicated for p-anisidine (kinact=2.85 min-1; KI=1.0 mM) and cyclopropyl bromide (kinact=4.4 min-1; KI=97 microM), but not for any of the remaining five inactivators, including acetylene. Ammonia slowed the rate of inactivation for acetylene and cyclopropyl bromide, but enhanced the rate of inactivation for the remaining inactivators. All seven compounds appear to be mechanism-based inactivators of AMO.

Ammonia↗

Identification of major subgroups of ammonia-oxidizing bacteria in environmental samples by T-RFLP analysis of amoA PCR products.

A cloning-independent method based on T-RFLP (terminal restriction fragment length polymorphism) analysis of amoA PCR products was developed to identify major subgroups of autotrophic ammonia oxidizers of the beta-subclass of the class Proteobacteria in total community DNA. Based on a database of 28 partial gene sequences encoding the active-site polypeptide of ammonia monooxygenase (amoA), defined lengths of terminal restriction fragments (= operational taxonomic units, OTUs) of amoA were predicted to correlate in TaqI-based T-RFLP analysis with phylogenetically defined subgroups of ammonia oxidizers. Members of the genus Nitrosospira showed a specific OTU of 283 bp in length, while a fragment size of 219 bp was indicative of Nitrosomonas-like sequence types including N. europaea, N. eutropha, and N. halophila. Two amoA sequence clusters designated previously as the lineages 'PluBsee' and 'Schöhsee' [Rotthauwe, J.-H., Witzel, K.-P., Liesack, W., 1997. Appl. Environ. Microbiol. 63, 4704-4712] shared a TaqI-based OTU with a fragment size of 48 bp, but sequence types of these two lineages could be differentiated by AluI-based T-RFLP analysis. A survey of various environmental samples and enrichment cultures by T-RFLP analysis and by comparative analysis of cloned amoA sequences confirmed the predicted correlations between distinct OTUs and phylogenetic information. Our data suggest that amoA-based T-RFLP analysis is a reliable tool to rapidly assess the complexity of ammonia-oxidizing communities in environmental samples with respect to the presence of major subgroups, i.e. nitrosospiras versus nitrosomonads.

Ammonia↗

Vasopressin accelerates experimental ammonia-induced brain edema in rats after portacaval anastomosis.

BACKGROUND/AIMS: Cerebral hyperemia is an important contributor to the development of brain edema in fulminant hepatic failure. Rats receiving an ammonia infusion after portacaval anastomosis (PCA) demonstrate a rise in cerebral blood flow (CBF) with brain edema at 180 min. Vasopressin (VP), a systemic vasoconstrictor which in the rat dilates cerebral vessels through V(2) receptors, was used to ascertain the effects of increasing CBF. METHODS: Changes in CBF were measured with Laser Doppler flowmetry (LDF). Absolute CBF was measured with radioactive microspheres to calculate oxygen and ammonia uptake. RESULTS: Compared to the NH(3)+Vehicle group, VP+NH(3) infusion accelerated the rise in CBF (117+/-21 vs. -6+/-12%, P<0.01), and the development of brain edema (81.09+/-0.17 vs. 80.29+/-0.06%, P<0.01). Radioactive microspheres confirmed these results (254+/-44 vs. 106+/-9.5 ml/min/100 g, P<0.01). Oxygen uptake was similar. Ammonia uptake was more than twofold higher in the VP+NH(3) group. A V(1) antagonist negated the higher mean arterial pressure (MAP) that occurs with VP but cerebral hyperemia still occurred. A V(2) antagonist resulted in similar systemic pressures, CBF and brain water compared to the VP+NH(3) group. CONCLUSIONS: In this model, an increase in CBF with VP hastens the development of brain edema while increasing ammonia delivery to the brain.

Ammonia↗

Ammonia prevents glutamate-induced but not low K(+)-induced apoptosis in cerebellar neurons in culture.

Cultured rat cerebellar granule neurons are widely used as a model system for studying neuronal apoptosis. Either low K(+) (5 mM) or low concentrations of glutamate (1-10 microM) induce apoptosis in cerebellar neurons in culture. However, the molecular mechanism(s) involved remain unclear. We show that long-term treatment with ammonia prevents glutamate-induced but not low K(+)-induced apoptosis in cerebellar neurons, as assessed by measuring DNA fragmentation and activation of caspase 3. Ammonia prevented glutamate-induced increase of intracellular calcium, depolarization of the inner mitochondrial membrane, release of cytochrome c to the cytosol, activation of caspase 3 and fragmentation of DNA. However, ammonia did not prevent low K(+)-induced activation of caspase 3 and fragmentation of DNA. These results indicate that the initial steps involved in the induction of apoptosis by low K(+) or by glutamate are different and that ammonia prevents glutamate-induced apoptosis by reducing glutamate-induced rise of intracellular Ca(2+), thus avoiding the activation of subsequent events of the apoptotic process.

Ammonia↗

Measurement of regional myocardial blood flow with N-13 ammonia and positron-emission tomography in intact dogs.

N-13 ammonia mimics certain properties of microspheres. It rapidly clears from blood into myocardium where it becomes fixed in proportion to myocardial blood flow. Used with positron emission tomography as a means for quantifying in vivo myocardial indicator concentrations, N-13 ammonia may be useful for noninvasive determination of myocardial blood flow with the arterial reference sampling technique. This possibility was examined in 27 experiments in 10 chronically instrumented dogs at control, high and low blood flows. Myocardial blood flow was calculated in vivo from the myocardial N-13 tissue activity concentrations derived from serial cross-sectional images of the heart, the 2 minute arterial input function and the withdrawal rate of arterial blood. These calculations were compared with blood flow determined by the standard microsphere technique. Blood flow determined in vivo with N-13 ammonia and positron emission tomography correlated with microsphere blood flow by y = -36.2 + 1.53x -0.0027x2 (r = 0.94 with a standard error of the estimate of 16 ml/min per 100 g). For flows from 44 to 200 ml/min per 100 g, the relation between in vivo and in vitro measured myocardial blood flow was nearly linear but reached a plateau at flows higher than 200 ml/min per 100 g. These results indicate that in dogs, blood flow in the physiologic range can be quantified in vivo with N-13 ammonia and positron emission tomography.

Ammonia↗

Assessment of diagnostic performance of quantitative flow measurements in normal subjects and patients with angiographically documented coronary artery disease by means of nitrogen-13 ammonia and positron emission tomography.

OBJECTIVES: Regional myocardial blood flow (MBF) and flow reserve measurements using nitrogen-13 (N-13) ammonia positron emission tomography (PET) were compared with quantitative coronary angiography to determine their utility in the detection of significant coronary artery disease (CAD). BACKGROUND: Dynamic PET protocols using N-13 ammonia allow regional quantification of MBF and flow reserve. To establish the diagnostic performance of this method, the sensitivity and specificity must be known for varying decision thresholds. METHODS: MBF and flow reserve for three coronary territories were determined in 20 normal subjects and 31 patients with angiographically documented CAD by means of dynamic PET and a three-compartment model for N-13 ammonia kinetics. Ten normal subjects defined the normal mean and SD of MBF and flow reserve, and 10 normal subjects were compared with patients. PET flow obtained in the territory with the most severe stenosis in each patient was correlated with the angiographic assessment of the stenosis (severity > or = 50%, > or = 70%, > or = 90%). Receiver operating characteristic (ROC) curve analysis was performed for 1.5, 2.0, 2.5, 3.0 and 4.0 SD of flow abnormalities. RESULTS: MBF and flow reserve values from the normal subjects and from territories with documented stenoses > or = 50% were significantly different (p < 0.05). A significant difference was found between normal subjects and angiographically normal territories of patients with CAD. High diagnostic accuracy and sensitivity, with moderately high specificity, were demonstrated for detection of all stenoses. CONCLUSIONS: Quantification of myocardial perfusion using dynamic PET and N-13 ammonia provides a high performance level for the detection and localization of CAD. The specificity of dynamic PET was excellent in patients with a low likelihood of CAD, whereas an abnormal flow reserve in angiographically normal territories was postulated to represent early functional abnormalities of vascular reactivity.

Adult↗

Cross-talk and ammonia channeling between active centers in the unexpected domain arrangement of glutamate synthase.

INTRODUCTION: The complex iron-sulfur flavoprotein glutamate synthase catalyses the reductive synthesis of L-glutamate from 2-oxoglutarate and L-glutamine, a reaction in the plant and bacterial pathway for ammonia assimilation. The enzyme functions through three distinct active centers carrying out L-glutamine hydrolysis, conversion of 2-oxoglutarate into L-glutamate, and electron uptake from an electron donor. RESULTS: The 3.0 A crystal structure of the dimeric 324 kDa core protein of a bacterial glutamate synthase was solved by the MAD method, using the very weak anomalous signal of the two 3Fe-4S clusters present in the asymmetric unit. The 1,472 amino acids of the monomer fold into a four-domain architecture. The two catalytic domains have canonical Ntn-amidotransferase and FMN binding (beta/alpha)8 barrel folds, respectively. The other two domains have an unusual "cut (beta/alpha)8 barrel" topology and an unexpected novel beta-helix structure. Channeling of the ammonia intermediate is brought about by an internal tunnel of 31 A length, which runs from the site of L-glutamine hydrolysis to the site of L-glutamate synthesis. CONCLUSIONS: The outstanding property of glutamate synthase is the ability to coordinate the activity of its various functional sites to avoid wasteful consumption of L-glutamine. The structure reveals two polypeptide segments that connect the catalytic centers and embed the ammonia tunnel, thus being ideally suited to function in interdomain signaling. Depending on the enzyme redox and ligation states, these signal-transducing elements may affect the active site geometry and control ammonia diffusion through a gating mechanism.

Ammonia↗

Evaluation of regional myocardial perfusion in patients with severe left ventricular dysfunction: comparison of 13N-ammonia PET and 99mTc sestamibi SPECT.

OBJECTIVE: Positron emission tomography (PET) scanning with 13N-ammonia and 18FDG is well established for the detection of myocardial viability. Due to the limited availability of PET facilities, recent studies have combined technetium 99m sestamibi single photon emission computed tomography (SPECT) with 18FDG PET or 18FDG SPECT. This approach enables simultaneous assessment of regional myocardial blood flow and metabolism and substantially increases the capacity for viability detection. To validate whether 99mTc-Sestamibi SPECT can replace 13N-ammonia PET, we compared these two modalities in patients with severe left ventricular dysfunction due to coronary artery disease. MATERIALS AND METHODS: Thirty-one patients (mean age 57+/-8 years; mean ejection fraction 27%+/-8%) with angiographically verified coronary artery disease were included. In random order, ammonia-PET and sestamibi-SPECT scans were performed. In a 20-segment model of the left ventricle, two blinded observers scored a total of 610 segments on a five-point scale. In a subset of 20 patients, 400 segments were scored twice to evaluate the observer variations of the two techniques. Segmental score differences were used to compare the imaging modalities. The impact on viability detection was assessed by combining the two flow tracers with FDG PET. RESULTS: Segmental comparison of the PET and SPECT studies yielded similar (difference < or = 1) results in 74% of segments, reflecting regional concordance values in the lateral, apical, anterior, septal, and inferior myocardial walls of 86%, 82%, 71%, 66%, and 63%, respectively. The differences in the septal and inferior walls were primarily due to overestimation of perfusion defects by sestamibi SPECT, which yielded a higher proportion of mismatch patterns in those regions. The overall observer variations of the PET and SPECT studies were 7.5% and 5.8%. CONCLUSION: Myocardial perfusion imaging with 13N-ammonia PET and 99mTc-sestamibi SPECT yielded similar results in patients with severe left ventricular dysfunction, except for the septal and inferior regions. In these regions, SPECT tended to overestimate perfusion defects. Hence, attenuation correction should be considered when combining FDG PET and sestamibi SPECT for diagnosing myocardial viability to avoid overestimation of mismatch patterns in those regions.

Adult↗