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PubMed2002

Reduction of sulfide, ammonia compounds, and adhesion properties of Lactobacillus casei strain KE99 in vitro.

The ability of Lactobacillus casei strain KE99 to reduce sulfide, ammonia, and to adhere to bio-surfaces was characterized and compared with three lactobacillus reference strains. Sulfide reduction by strain KE99 in MRS broth increased exponentially after 10-h growth and reached a maximum (>300 ppm reduction) within 48 h. KE99 demonstrated a maximum reduction of sulfide under anaerobic (341 ppm) growth conditions at pH 6.0-8.0 range. Maximum anaerobic reduction of sulfide was demonstrated by L. casei 393 at pH 7.0 (272 ppm); L. rhamnosus at pH 8.0 (277 ppm); and L. reuteri at pH 7.0 (244 ppm). KE99 reduced sulfide more (p < 0.0001) in MRS broth spiked with Na2S (374 ppm) than (NH4)2S (340 ppm) salts. Ammonia reduction by strain KE99 and the three lactobacillus reference strains in MRS broth was low. Ammonia reduction reached a maximum within 36 h and remained unchanged over extended incubations of 48 h to 72 h or further. KE99 reduced ammonium sulfate (37 ppm) more readily than the nitrate (31 ppm), hypophosphate (29 ppm), or chloride (20 ppm) salts of ammonia. KE99 and the three reference strains of lactobacilli demonstrated avid binding to Bio-coat (Cn type-I, Cn type-IV, laminin, fibronectin), Matrigel, and Caco-2 cell monolayers in vitro. The number of lactobacilli binding to Caco-2 was estimated at 74/cell with strain KE99, which was significantly higher compared with 40/cell (p < 0.0001), 26/cell (0.0001), and 64/cell (p < 0.002) with L. casei 393, L. reuteri, and L. rhamnosus, respectively. The interaction of KE99 to immobilized Cn type-I was saturable and reached an equilibrium within 1 h at room temperature. KE99 binding to Cn type-I occurred at a wide pH range and was biphasic with maximum binding at pH 5.5 and 7.5. Inhibition and binding-displacement experiments with different salts and sugars suggested that the KE99 binding to immobilized Cn type-I may involve a combination of electrostatic and lectin-type interactions. KE99 effectively detached the Cn-adherent E. coli O157:H7 in the range of 55% (ATCC43895) to 76% (ATCC43894). The binding-displacement values for L. casei 393, L. reuteri and L. rhamnosus to detach Cn-adherent E. coli O157:H7 (ATCC43894) were 66 +/- 4%, 59 +/- 2%, and 64 +/- 2%, respectively. Also, a reconstituted solution of the freeze-dried KE99 preparation effectively detached the Cn-adherent E. coli O157:H7 in a dose-dependent manner that reached a binding-displacement equilibrium of 85% at a 1% wt/vol KE99 concentration.

Ammonia↗
PubMed2005

Characterization of transgenic poplar with ectopic expression of pine cytosolic glutamine synthetase under conditions of varying nitrogen availability.

The present study addresses the hypothesis that enhanced expression of glutamine synthetase (GS) in transgenic poplar, characterized by the ectopic expression of pine cytosolic GS, results in an enhanced efficiency of nitrogen (N) assimilation and enhanced growth. Transgenic and control poplar were supplied with low and high N levels and the role of ectopic expression of the pine GS in growth and N assimilation was assessed by using amino acid analysis, (15)N enrichment, biochemical analyses, and growth measurements. While leaves of transgenic poplar contained 85% less (P < 0.01) free ammonium than leaves of nontransgenic control plants, leaves of transgenics showed increases in the levels of free glutamine and total free amino acids. Transgenic poplar lines also displayed significant increases in growth parameters when compared with controls grown under both low (0.3 mm) and high (10 mm) nitrate conditions. Furthermore, (15)N-enrichment experiments showed that 27% more (P < 0.05) (15)N was incorporated into structural compounds in transgenic lines than in nontransgenic controls. Using the methods described here, we present direct evidence for increased N assimilation efficiency and growth in GS transgenic lines.

Amino Acids↗
PubMed1977

H2 metabolism in the photosynthetic bacterium Rhodopseudomonas capsulata: H2 production by growing cultures.

Purple photosynthetic bacteria produce H2 from organic compounds by an anaerobic light-dependent electron transfer process in which nitrogenase functions as the terminal catalyst. It has been established that the H2-evolving function of nitrogenase is inhibited by N2 and ammonium salts, and is maximally expressed in cells growing photoheterotrophically with certain amino acids as sources of nitrogen. In the present studies with Rhodopseudomonas capsulata, nutritional factors affecting the rate and magnitude of H2 photoproduction in cultures growing with amino acid nitrogen sources were examined. The highest H2 yields and rates of formation were observed with the organic acids: lactate, pyruvate, malate, and succinate in media containing glutamate as the N source; under optimal conditions with excess lactate, H2 was produced at rates of ca. 130 ml/h per g(dry weight) of cells. Hydrogen production is significantly influenced by the N/C ratio in the growth substrates; when this ratio exceeds a critical value, free ammonia appears in the medium and H2 is not evolved. In the "standard" lactate + glutamate system, both H2 production and growth are "saturated" at a light intesity of ca. 600 ft-c (6,500 lux). Evolution of H2, however, occurs during growth at lithe intensities as low as 50 to 100 ft-c (540 to 1,080 lux), i.e., under conditions of energy limitation. In circumstances in which energy conversion rate and supplies of reducing power exceed the capacity of the biosynthetic machinery, energy-dependent H2 production presumably represents a regulatory device that facilitates "energy-idling." It appears that even when light intensity (energy) is limiting, a significant fraction of the available reducing power and adenosine 5'-triphosphate is diverted to nitrogenase, resulting in H2 formation and a bioenergetic burden to the cell.

Amino Acids↗
PubMed2003

Nitrogen isotope analysis of ammonium in aqueous solutions using a perfluorosulfonated ionomer membrane for solid-phase microextraction.

The use of custom-made solid-phase microextraction (SPME) fibers coated with a perfluorosulfonated ionomer, Nafion, was investigated for nitrogen isotopic analysis of ammonium in aqueous solutions. Aqueous ammonium was converted to ammonia by addition of a base, followed by absorption from the headspace, desorption in the injection port of a gas chromatograph, and analysis by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). Fibers coated with a Nafion tubing were chosen due to a higher fiber-gas distribution constant and a higher Nafion thickness than fibers coated with Nafion solution, both leading to a higher amount of ammonia absorbed at equilibrium. The Nafion membrane-coated fiber absorbed approximately 20 times more than a commercial polydimethylsiloxane (PDMS) fiber. The isotopic fractionation between fiber and gas was 1.0117 +/- 0.0009 (standard deviation, SD, of all measurements) at an initial ammonia gas concentration of 21-210 microM. At 390 microM initial gas concentration it was slightly lower. When sampling from liquid samples, an ammonium concentration of 10 mM was needed to obtain a sufficient amount of ammonia absorbed. Modeling of the absorption at different temperatures showed that the absorption was approximately constant in the temperature range suitable for SPME experiments. Absorption at room temperature was therefore used for simplicity. A pilot study was conducted in which absorption was achieved from a single 9 microL droplet of sample. The preliminary results showed that delta(15)N analysis was possible for only 0.4-0.5 micromol of ammonium with a SD of 0.8 per thousand (n = 5).

Algorithms↗
PubMed1977

Utilization of nitrogen compounds and ammonia assimilation by Chromatiaceae.

Chromatium vinosum strain D, Thiocapsa roseopersicina strain 6311 and Ectothiorhodospira mobilis strain 8112 were grown anaerobically in the light with various single nitrogen sources. When substituted for NH4Cl only glutamine and casamino acids supported good growth of all strains tested. Peptone and urea were utilized by C. vinosum and T. roseopersicina, glutamate, asparagine and nitrate only by C. vinosum. The strains were able to grow with molecular nitrogen; complete inhibition of this growth was observed in the presence of alanine with E. mobilis, and of alanine or asparagine with T. roseopersicina. Glutamate dehydrogenase, requiring either NADH or NADPH, NADH-linked glutamate synthase, and glutamine synthetase were demonstrated in the above organisms grown on NH4Cl.

Amino Acids↗
PubMed1998

Use of soybean oil and ammonium sulfate additions to optimize secondary metabolite production.

A valine-overproducing mutant (MA7040, Streptomyces hygroscopicus) was found to produce 1.5 to 2.0 g/L of the immunoregulant, L-683,590, at the 0.6 m3 fermentation scale in a simple batch process using soybean oil and ammonium sulfate-based GYG5 medium. Levels of both lower (L-683,795) and higher (HH1 and HH2) undesirable homolog levels were controlled adequately. This batch process was utilized to produce broth economically at the 19 m3 fermentation scale. Material of acceptable purity was obtained without the multiple pure crystallizations previously required for an earlier culture, MA6678, requiring valine supplementation for impurity control. Investigations at the 0.6 m3 fermentation scale were conducted, varying agitation, pH, initial soybean oil/ammonium sulfate charges, and initial aeration rate to further improve growth and productivity. Mid-cycle ammonia levels and lipase activity appeared to have an important role. Using mid-cycle soybean oil additions, a titer of 2.3 g/L of L-683,590 was obtained, while titers reached 2.7 g/L using mid-cycle soybean oil and ammonium sulfate additions. Both higher and lower homolog levels remained acceptable during this fed-batch process. Optimal timing of mid-cycle oil and ammonium sulfate additions was considered a critical factor to further titer improvements.

Ammonium Sulfate↗
PubMed1985

Effects of ammonium salts on synaptic transmission to hippocampal CA1 and CA3 pyramidal cells in vivo.

The effects of ammonium acetate or chloride, perfused through the lateral ventricle, were studied on the hippocampal formation of the rat. During perfusion with ammonia, the population spikes, evoked by stimuli delivered to the fimbria, were first increased and then reduced. On the other hand, the late positive wave gradually decreased throughout the application of ammonia. The inhibition, studied by the paired-pulse test, was found to be reduced when the population spike was transiently enhanced, indicating that disinhibition could be responsible for the enhancement of synaptically evoked responses. Neither antidromically evoked population spikes nor the typical effects of iontophoretically applied glutamate, aspartate or gamma-aminobutyrate were changed by ammonia. These findings can be accounted for by a single action of ammonia, a depression of excitatory synaptic transmission, the excitatory synapses on inhibitory interneurons being more readily depressed than those on the pyramidal cells. Both effects, early hyperexcitability and late depression, are probably due to a reduction in the release of the excitatory neurotransmitter, glutamate and/or aspartate. We tentatively suggest that these mechanisms are responsible for some of the symptoms observed during the development of hyperammonemic encephalopathies.

Acetates↗
PubMed1992

Choline transport in Fusarium graminearum A 3/5.

Fusarium graminearum A 3/5 possesses a high affinity system (Km = 32 +/- 8 microM; mean +/- SE) for uptake of choline, which was shown to be energy-dependent and constitutive. The maximum rate of choline uptake by this system was repressed by ammonia and glucose, showing a three-fold increase in maximum activity after nitrogen (2 h) or carbon (4 h) starvation. The system was highly specific for choline with only dimethylethanolamine (Ki = 198 +/- 29 microM), betaine aldehyde (Ki = 95 +/- 14 microM) and chlorocholine (Ki = 352 +/- 40 microM) acting as competitive inhibitors. Hemicholinium-3 acted as a mixed (non-competitive) inhibitor (KIES = 1.9 +/- 0.6 microM; KIE = 3.6 +/- 1.9 microM).

Biological Transport, Active↗
PubMed2006

Simultaneous removal of phenol, ammonium and thiocyanate from coke wastewater by aerobic biodegradation.

A laboratory-scale activated sludge plant composed of a 20 L volume aerobic reactor followed by a 12 L volume settling tank and operating at 35 degrees C was used to study the biodegradation of coke wastewater. The concentrations of ammonium nitrogen (NH(4)(+) -N), phenols, chemical oxygen demand (COD) and thiocyanate (SCN(-)) in the wastewater ranged between 504 and 2,340, 110 and 350, 807 and 3,275 and 185 and 370 mg/L, respectively. The study was undertaken with and without the addition of bicarbonate. The addition of this inorganic carbon source was necessary to favour nitrification, as the alkalinity of the wastewater was very low. Maximum removal efficiencies of 75%, 98% and 90% were obtained for COD, phenols and thyocianates, respectively, without the addition of bicarbonate. The concentration of ammonia increased in the effluent due to both the formation of NH(4)(+) as a result of SCN(-) biodegradation and to organic nitrogen oxidation. A maximum nitrification efficiency of 71% was achieved when bicarbonate was added, the removals of COD and phenols being almost similar to those obtained in the absence of nitrification. Batch experiments were performed to study the influence of pH and alkalinity on the biodegradation of phenols and thiocyanate.

Aerobiosis↗
PubMed1998

Water purification system by using the biofilter for long-term experiment equipment with aquatic animals for the space station.

We have developed a water purification system that enables long-term experiment with aquatic animals for 90 days or more on the space station. We designed the system that combined a biofilter for ammonia removal (nitrification) with another for nitrate removal (denitrification). The experiment with goldfish was for 90 days with an aquatic animals' examination device. The equipment consists of a fish tank, a filter module, pumps, and an artificial lung for gas exchange. The goldfish were kept in the tank without any water replacement throughout the experiment. When a filter module consists of adsorbents without bacteria, the concentration of the nitrite and ammonia begin to increase so that the goldfish die. On the contrary, neither ammonia nor nitrite accumulated throughout the experiment, and the concentration of T-N also maintained 30 ppm or less when the combined biofilter was used. Moreover, no fish died throughout the period. The water purification system with biofilter enabled us to examine the long-term life support testing. We also report a new denitrification (correction of dentrification) method for the life support system.

Animals↗
PubMedDate not supplied

[Glutamine metabolism in the brain and in the liver in critical conditions].

Experiments on cats and rats have established that critical conditions caused by acute hemorrhage, hepatotoxin, and hepatectomy lead to ammonia accumulation in the brain and liver due to the predominance of decay of glutamine over its formation in these organs. With this, the depressed formation of glutamine is a universal cell response to a pathogenic agent whereas a change in glutamine deamination in disease depends on both the nature of a pathogenic agent and the organ wherein this reaction occurs.

Animals↗
PubMed2005

Determination of ammonium in river water and sewage samples by capillary zone electrophoresis with direct UV detection.

We developed capillary zone electrophoresis (CZE) with direct UV detection for determination of ammonium in environmental water samples. Ammonium in the samples was partly converted into ammonia in the alkaline background electrolyte (BGE) during migration and was detected by molecular absorption of ammonia at 190 nm in approximately 7 min. The limit of detection (LOD) for ammonium was 0.24 mg/l (as nitrogen) at a signal-to-noise ratio of three. The respective values of the relative standard deviation (RSD) of peak area, peak height, and migration time for ammonium were 2.1, 1.8, and 0.46%. Major alkali and alkaline earth metal ions coexisting in the samples did not interfere with ammonium determination by the proposed method. The proposed method determined ammonium in surface water and sewage samples. The results were compared to those obtained using ion chromatography (IC).

Calibration↗
PubMed1988

Protection against acute hyperammonemia: the role of quaternary amines.

The quaternary amine L-carnitine is able to protect Swiss Albino mice from hyperammonemia when administered in high doses before ammonium acetate. This has been explained by its specific ability to shuttle fatty acids into mitochondria. The structure of L-carnitine resembles the chemical structure of other substances that have been described as being able to protect living cells against osmotic stress. We subjected Swiss Albino mice to hyperammonemia after pretreatment with L-carnitine or "osmoprotectants" such as the quaternary amines choline and betaine, and trimethylamine N-oxide. L-Carnitine proved to be the drug of choice to protect against acute hyperammonemia. Nevertheless, the other tested compounds appeared also to be effective, suggesting that osmoregulation plays a major role in protection against hyperammonemia.

Acetates↗
PubMed1997

A Neurospora crassa mutant altered in the regulation of L-amino acid oxidase.

The isolation and characterization of a Neurospora crassa mutant altered in L-amino oxidase regulation is reported. The previously isolated gln-1bR8 strain, which only synthesizes the glutamine synthetase alpha monomer and lacks the beta monomer, was used as parental strain. A mutant derivative of strain was selected for its ability to grow on minimal medium in the presence of DL-methionine-SR-sulfoximine (MSO), an inhibitor of glutamine synthetase activity. This gln-1bR8;MSOR mutant overcame the inhibitory effect of MSO by increasing the activity of L-amino acid oxidase, an enzyme capable of degrading this compound. In contrast with the wild-type strain, the L-amino acid oxidase of the MSOR mutant was resistant to glutamine repression; in fact, it was induced by this amino acid but repressed by ammonium. This mutant is different from other nitrogen regulatory N. crassa mutants reported and is only altered in the regulation of L-amino acid oxidase. The MSOR mutation is epistatic to nit-2 since the nit2;MSOR double mutant regulated the L-amino acid oxidase in the same way as the MSOR single mutant.

Amino Acid Oxidoreductases↗
PubMed2001

Intensely positively charged perineuronal nets in the adult rat brain as detected by staining with anionic iron colloid.

Ferric chloride, when boiled with ammonium thiocyanate, ammonia and cacodylic acid, is converted into a fine anionic iron colloid which consists of 1.0-1.5 nm electron dense granules and gives a distinct Prussian blue reaction (OHTSUKA and MURAKAMI, 1986). Light microscopy of tissue sections stained with this fine anionic iron colloid at pH values of 6.0, 7.0 and 8.0 showed that the healthy adult rat brain contains a considerable number of neurons which possess an intensely positively charged perineuronal net. This net was most clearly demonstrable by staining with the anionic iron colloid at a pH value of 8.0, at which ionizations of almost all cationic sites of the tissue elements were obliterated. Transmission electron microscopy of ultrathin sections stained at a pH value of 8.0 showed that the anionic iron colloid was preferentially deposited in the perineuronal tissue spaces. These findings indicate that the intensely positively charged perineuronal net contains some strongly basic substances such as guanidino compounds, and occupies the perineuronal (perisynaptic) tissue space.

Animals↗
PubMed2001

Formation of gas-phase clusters monitored during electrospray mass spectrometry: a study of quaternary ammonium pesticides.

Gas-phase cluster formation between the quaternary ammonium pesticides paraquat, diquat, difenzoquat, chlormequat and mepiquat, and chloride and acetate anions present in a liquid chromatography (LC) mobile phase, has been studied using electrospray mass spectrometry. The clusters of paraquat, mepiquat and chlormequat were revealed over the entire m/z range of the mass spectrometer, and their formation is dependent on the concentrations of both the cationic and the anionic species. Mepiquat and chlormequat form clusters of the type [2M(q)(+) + A(-)](+), where M(q)(+) is the quaternary ammonium cation and A(-) is the anion. Paraquat forms a cluster species with ammonia and also an ion-pair complex with chloride anions. Diquat and difenzoquat did not form observable ion-pair complexes or clusters with any of the anions studied. Competitive binding of acetate and chloride anions reflects the higher charge density of chloride, which forms the dominant clusters with mepiquat and chlormequat. The formation of cluster species has implications for the quantification of quaternary ammonium pesticides and may have an influence on the linearity of calibrations.

Acetates↗
PubMed2002

Change of mobile phase pH during gradient reversed-phase chromatography with 2,2,2-trifluoroethanol-water as mobile phase and its effect on the chromatographic hydrophobicity index determination.

We have shown previously that using a trifluoroethanol containing mobile phase provides a unique chromatographic selectivity. This is essential to derive molecular descriptors by HPLC which requires retention data from several systems. It also requires that the ionisation is suppressed so that retention times reflect the properties of the neutral molecules. Therefore the pH change of the mobile phase during gradient elution and its effect on the solute ionisation have been studied. During gradient elution of mixtures of ammonium acetate and butylammonium formate with trifluoroethanol as an organic modifier it was found that the pH was almost constant when the gradient started with a low pH. However, when the starting mobile phase pH was above 8 the pH dropped very quickly as the trifluoroethanol concentration increased in the mobile phase. The CHI descriptor (a retention index derived directly from gradient retention times) of several basic compounds as a function of starting mobile phase pH has been measured using trifluoroethanol gradient. The effect of the trifluoroethanol on the pKa change of the compounds has been investigated. The experimental data fit closely to a previously derived equation that describes gradient retention times as a function of mobile phase pH and analyte ionisation constant (pKa). This equation makes it possible to predict the CHI descriptor for ionisable compounds at various pH values. We have used butylamine for high pH mobile phase preparation as is more basic than ammonia and for many basic drugs the retention of the neutral form could be obtained directly (without extrapolation).

Buffers↗
PubMed1999

An evaluated improvement of the extinction dilution method for isolation of ammonia-oxidizing bacteria.

An improved method for isolation of ammonia-oxidizing bacteria (AOB) by the extinction dilution technique is described. It is important to prevent the growth of heterotrophic organisms, which may easily outnumber the AOB in mixed cultures. This was achieved by careful elimination of C sources in the medium and by sealing the cultures from contact with the atmosphere, thus excluding air-borne, volatile compounds which support growth of heterotrophs. The sealing of the cultures reduced the number of heterotrophs by a factor of 10, thus grossly increasing the chances of obtaining pure AOB cultures. Another important factor is to use actively growing 'late log' cultures during the final isolation step. This was achieved by adjusting the buffer capacity to ensure a clearly visible pH indicator shift at a stage when one-third to one-half of the ammonia had been oxidized. By this improved isolation procedure, AOB were isolated from three different locations: an arable soil, a lead-contaminated soil and an animal house. For an unknown reason, several attempts to isolate pure cultures from a forest soil were unsuccessful, despite the presence of AOB in the primary extinction dilution cultures. The isolates from soils were all Nitrosospira spp. For isolation of AOB from the animal house, two growth media were used, one containing ammonium sulfate, and one containing urea. From the cultures with ammonium sulfate, Nitrosomonas spp. were isolated, whereas Nitrosospira spp. were isolated from the cultures with urea as the main ammonia source. The identifications of all isolates are based on morphology and 16S rDNA sequences.

Ammonia↗