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At least 811 records · Page 45Linked to original sources
PubMed1970

Correlation of ultrastructure in Azotobacter vinelandii with nitrogen source for growth.

Azotobacter synthesizes an extensive internal membranous nework when grown with air (N(2)), i.e., under conditions when these bacteria fix nitrogen. Very slight quantities of internal membrane, concentrated mainly about the cell periphery, are formed when Azotobacter grows with fixed nitrogen, i.e., ammonia and amino acids. Compared to cells growing with ammonia, cells utilizing atmospheric nitrogen as the sole nitrogen source are smaller in size and volume, grow one-third slower, and lack detectable poly-beta-hydroxybutyrate.

Air↗
PubMed2005

Experimental generation and computational modeling of intracellular pH gradients in cardiac myocytes.

It is often assumed that pH(i) is spatially uniform within cells. A double-barreled microperfusion system was used to apply solutions of weak acid (acetic acid, CO(2)) or base (ammonia) to localized regions of an isolated ventricular myocyte (guinea pig). A stable, longitudinal pH(i) gradient (up to 1 pH(i) unit) was observed (using confocal imaging of SNARF-1 fluorescence). Changing the fractional exposure of the cell to weak acid/base altered the gradient, as did changing the concentration and type of weak acid/base applied. A diffusion-reaction computational model accurately simulated this behavior of pH(i). The model assumes that H(i)(+) movement occurs via diffusive shuttling on mobile buffers, with little free H(+) diffusion. The average diffusion constant for mobile buffer was estimated as 33 x 10(-7) cm(2)/s, consistent with an apparent H(i)(+) diffusion coefficient, D(H)(app), of 14.4 x 10(-7) cm(2)/s (at pH(i) 7.07), a value two orders of magnitude lower than for H(+) ions in water but similar to that estimated recently from local acid injection via a cell-attached glass micropipette. We conclude that, because H(i)(+) mobility is so low, an extracellular concentration gradient of permeant weak acid readily induces pH(i) nonuniformity. Similar concentration gradients for weak acid (e.g., CO(2)) occur across border zones during regional myocardial ischemia, raising the possibility of steep pH(i) gradients within the heart under some pathophysiological conditions.

Acetates↗
PubMed1975

Analysis of regulatory factors for urea synthesis by isolated perfused rat liver. I. Urea synthesis with ammonia and glutamine as nitrogen sources.

Urea synthesis was studied using the isolated liver perfusion with ammonium cholride and glutamine as nitrogen sources. The rate of urea formation increases with ammonium cholorde concentration up to 5mM, and the rate remained constant in the range between 5 and 20mM of ammonium chloride as the substrate. The concentration of ammonia in the medium to support the half-maximum velocity of urea formation was 0.7mM. The rate of urea formation was stimulated by the addition of 2.5mM ornithine, and the greater part of the ornithine which was taken up into the liver was accumulated as citrulline in the presence of ammonia. A considerable accelerating effect of N-acetylglutamate on the synthetic rate was observed, but a rather high concentration of N-acetylglutamate was required in order to obtain the maximum effect possibly, because its permeability into liver cells may be limited. A marked additive effect on the rate of urea formation was observed with the combined addition of ornithine and N-acetylglutamate. The metabolic conversion of glutamine nitrogen to urea in the perfused rat liver and the effect of several compounds which stimulated urea synthesis with ammonia were further examined. The process of conversion of glutamine nitrogen to urea might be composed of the following three steps. In the first lag phase, a small amount of glutamine was removed from the medium. In the second stage, the glutamine level decreased rapidly and ammonia was accumulated in the perfusate. The third stage was a period in which glutamine concentration remained at a constant low level, and the accumulated ammonia was rapidly conversed to urea. The rate of urea formation in this third stage was found to be much higher than that with ammonia as the substrate. The maximum rate of glutamine removal was obtained at pH 7.7 of the perfusate and at a concentration of 10mM glutamine. Urea formation with glutamine was also stimulated by the addition of ornithine, malate, or N-acetylglutamate, which had accelerating effects on the urea synthesis with ammonia. This stimulation was due to an effective conversion of ammonia to urea, but no change in the rate of removal glutamine was obtained.

Ammonium Chloride↗
PubMed2006

Retention of ionisable compounds on high-performance liquid chromatography XVII. Estimation of the pH variation of aqueous buffers with the change of the methanol fraction of the mobile phase.

The use of methanol-aqueous buffer mobile phases in HPLC is a common election when performing chromatographic separations of ionisable analytes. The addition of methanol to the aqueous buffer to prepare such a mobile phase changes the buffer capacity and the pH of the solution. In the present work, the variation of these buffer properties is studied for acetic acid-acetate, phosphoric acid-dihydrogenphosphate-hydrogenphosphate, citric acid-dihydrogencitrate-hydrogencitrate-citrate, and ammonium-ammonia buffers. It is well established that the pH change of the buffers depends on the initial concentration and aqueous pH of the buffer, on the percentage of methanol added, and on the particular buffer used. The proposed equations allow the pH estimation of methanol-water buffered mobile phases up to 80% in volume of organic modifier from initial aqueous buffer pH and buffer concentration (before adding methanol) between 0.001 and 0.01 mol L(-1). From both the estimated pH values of the mobile phase and the estimated pKa of the ionisable analytes, it is possible to predict the degree of ionisation of the analytes and therefore, the interpretation of acid-base analytes behaviour in a particular methanol-water buffered mobile phase.

Chromatography, High Pressure Liquid↗
PubMed1994

Determination of ammonium and L-glutamine in hybridoma cell cultures by sequential flow injection analysis.

A flow injection analytical system based on a gas diffusion membrane module for ammonia and an ammonium flow-through potentiometric detector has been set up for measurement of L-glutamine and ammonium ions in hybridoma cell cultures. The main feature of the system is that the same basic analytical concept and equipment is used in both measurements, the only difference being for the determination of L-glutamine, in which the sample flows through an immobilized glutaminase cartridge. The conditions to enable the performance of both analysis consecutively, avoiding potential interferences by unwanted deamination of other compounds in the samples, have been determined. Finally, the proposed system has been compared with reference analytical methods for batch hybridoma cell culture experiments.

Animals↗
PubMed2004

Retention of ionisable compounds on high-performance liquid chromatography. XV. Estimation of the pH variation of aqueous buffers with the change of the acetonitrile fraction of the mobile phase.

The most commonly used mobile phases in reversed-phase high-performance liquid chromatography (RP-HPLC) are hydro-organic mixtures of an aqueous buffer and an organic modifier. The addition of this organic solvent to buffered aqueous solutions involves a variation of the buffer properties (pH and buffer capacity). In this paper, the pH variation is studied for acetic acid-acetate, phosphoric acid-dihydrogenphosphate-hydrogenphosphate, citric acid-dihydrogencitrate-citrate, and ammonium-ammonia buffers. The proposed equations allow pH estimation of acetonitrile-water buffered mobile phases up to 60% (v/v) of organic modifier and initial aqueous buffer concentrations between 0.001 and 0.1 mol L(-1), from the initial aqueous pH. The estimated pH variation of the mobile phase and the pKa variation of the analytes allow us to predict the degree of ionisation of the analytes and from this and analyte hydrophobicities, to interpret the relative retention and separation of analyte mixtures.

Acetonitriles↗
PubMed2005

Preparation and evaluation of ammonia-treated collagen/chitosan matrices for liver tissue engineering.

To further enhance the properties of existing collagen/chitosan scaffolds for liver tissue engineering, a very simple method was developed to form noncovalently linked mimic of the liver extracellular matrices. Collagen/chitosan mixtures in various proportions (i.e., 1:0, 3:2, 1:1, 2:3, and 0:1 v/v) were lyophilized or evaporated to form sponges or flat films before they were gelled using an aqueous 25% ammonia solution. The porosities of the obtained sponges were above 90% with various pore sizes. The highest mechanical strength (1.9+/-0.7 MPa) and the lowest degradation time (65+/-1.7 days) were achieved by the collagen/chitosan (1:1) matrices. Hepatocytes cultured on the collagen/chitosan (1:1) matrices exhibited relatively high glutamate-oxaloacetate transaminase and glucose secretion functions 25 days post-seeding. Nuclues of the hepatocytes were more elongated and arranged in certain directions on the 1:1 matrices. The cytocompatibility and enhanced biostability of our new ammonia-treated collagen/chitosan matrices suggest that they could be used as scaffolds for liver tissue engineering.

Amines↗
PubMed2001

Water-soluble luminal contents of the gut of the earthworm Lumbricus terrestris L. and their physiological significance.

In the post-gizzard gut of the earthworm Lumbricus terrestris, distinguishing the functions of the luminal epithelium from those of the chloragogenous tissue has been hindered by the close apposition of these two tissues. Moreover, both tissues may have different functions from the anterior to the posterior of the animal. We analyzed the gut luminal contents of L. terrestris so as to gain a better understanding of the function of the luminal epithelium. The intestine was divided into four regions from anterior to posterior, and the water-soluble portion of the luminal contents of these four regions was analyzed for protease and amylase activity, calcium and ammonium ions, and protein. The same four regions of the gut wall were analyzed for glutamate dehydrogenase (GDH) and serine dehydratase (SDH) to determine their location with reference to the site of ammonia production. We observed high levels of proteases, amylase, protein and calcium ions in the gut luminal contents of the first two regions, and a significant decline of all four variables in region III. Conversely, ammonia was low in the gut contents of regions I and II but rose sharply in region III, which was also the region to which the tissue enzymes GDH and SDH were localized. The ammonia content of earthworm casts was observed to be much higher than that of the surrounding soil. These data are presented as partial evidence for the proposal that the excretory ammonia produced by feeding earthworms is a product of the luminal epithelium of region III of the gut. It is also proposed that ammonia and calcium may function as ion-exchangers in the absorptive function of the earthworm gut.

Animals↗
PubMed2001

An experimental and theoretical study of the basicity of tetra-tert-butyltetrahedrane.

The gas-phase basicity (GB) of tetra-tert-butyltetrahedrane (tBu4THD) was determined by FT-ICR mass spectrometry and comparison with reference compounds of known basicity. Its GB, 1035+/-10 kJ x mol(-1), makes tetra-tert-butyltetrahedrane one of the strongest bases reported so far. Ab initio calculations [B3LYP/6-31G(d) and B3LYP/6-311 + G(d,p)//6-31G(d)] have been carried out in order to compare the high experimental basicity of tBu4THD with that estimated theoretically. Both B3LYP/6-31G(d) and QCISD(T) calculations were used to determine the reaction path which connects the initial tetrahedrane-ammonium complex with the final products, protonated cyclobutadiene (CBDH+) and ammonia.

Journal Article↗
PubMed1997

Evidence for two possible glnB-type genes in Herbaspirillum seropedicae.

Two glnB-like genes have been isolated from Herbaspirillum seropedicae by complementation of the Klebsiella pneumoniae glnB502 mutant for growth on nitrate. One of these glnB-like genes has been sequenced and shows strong identity with GlnB proteins derived from other organisms. A Tn5-20 mutation of this glnB was Nif negative.

Ammonium Chloride↗
PubMed1975

Nutritional control of differentiation (sclerotization) of the myxomycete Physarum flavicomum.

During differentiation (sclerotization) of the Myxomycete Physarum flavicomum, the acellular phasmodium converts into numerous dormant cells surrounded by cell walls. This work establishes that a condition of nutrient imbalance triggers the differentiation process. Specifically, the unavailability of an adequate spectrum of amino acids in the medium initiates the metabolic and morphological alterations characteristic of the sclerotizing plasmodium. In the absence of extracellular amino acids, the cellular pool of amino acids and cellular protein were catabolized as differentiation proceeded. The pattern of amino acids in the cellular pool also changed during differentiation, as the content of pool amino acids was reduced at least 75 percent. The decrease in protein content was negligible after 12 h incubation but was about 40 percent at 48 h when differentiation was complete. However, in the presence of extracellular amino acids, protein degradation, amino acid pool depletion, and differentiation were all inhibited. Ammonium ions (12.4 mM) similarly delayed differentiation. Differentiation, amino acid pool depletion, and the degradation of cellular protein readily occurred in the presence of an extracellular supply of dextrose, which stimulated cell wall formation. The effect of dimethyl sulfoxide, cyclic 3'-5'-adenosine monophosphate, glutathione, diamide, and other compounds on the differentiation process are reported also.

Amides↗
PubMed2000

Molecular evidence for genus level diversity of bacteria capable of catalyzing anaerobic ammonium oxidation.

Recently, a bacterium capable to oxidize ammonium anaerobically at a high rate was identified as novel member of the Planctomycetales (Strous, M., Fuersi, J. A., Kramer, E. H. M., Logemann, S., Muyzer, G., van de Pas-Schoonen, K. T., Webb, R. I., Kufnen, J. G., and Jetten, M. S. M.: Nature 400, 446-449, 1999). Here we investigated the microbial community structure of a trickling filter biofilm with a high anaerobic ammonium oxidation activity. Fluorescence in situ hybridization (FISH) with a set of nine probes designed for specific identification of the recently described anaerobic ammonium oxidizer demonstrated that only one probe hybridized to bacteria within the biofilm. For phylogenetic characterization of putative biofilm anaerobic ammonium oxidizers a full-cycle 16S rDNA approach was performed by using a Planctomycetales-specific forward primer for PCR amplification. Of the twenty-five 16S rDNA fragments (1364 bp in length) amplified from the biofilm, nine were affiliated to the Planctomycetales. Comparative analysis showed that these sequences were more than 98.9% similar to each other but only distantly related to the previously recognized anaerobic ammonium oxidizer (below 91% similarity) and all other organisms represented in public 16S rRNA databases (similarities of below 79%). The retrieved sequences and the previously recognized anaerobic ammonium oxidizer represent two well-separated groups of a deep-branching lineage within the Planctomycetales. Quantitative FISH analysis with a newly designed specific probe showed that the novel bacterium, provisionally classified as "Candidatus Kuenenia stuttgartiensis" constituted the dominant fraction of the biofilm bacteria. In situ probing revealed that ammonia-oxidizing bacteria of the beta-subclass of Proteobacteria were also present, albeit in significant smaller amounts, within the anoxic biofilm. Comparative sequence analysis of a stretch of the gene encoding ammonia-monooxygenase (amoA) demonstrated the occurrence of the DNA of at least three different populations of beta-subclass ammonia oxidizers within the biofilm.

Anaerobiosis↗
PubMed1992

Structure of pentaamminechlororuthenium(III) bisulfate tetrahydrate.

[RuCl(NH3)5](HSO4)2.4H2O, M(r) = 487.88, triclinic, P1, a = 10.0422 (5), b = 14.1044 (7), c = 6.3273 (6) A, alpha = 100.369 (5), beta = 98.655 (6), gamma = 81.354 (4) degree, V = 864.5 (3) A3, Z = 2, Dm = 1.86 (1), D chi = 1.874 g cm-3, lambda(MO K alpha) = 0.71073 A, mu = 13.3 cm-1, F(000) = 498, T = 296 (1) K, R = 0.025 for 4622 unique reflections with I greater than 3 sigma(I). The structure contains (NH3)5RuCl2+ cations, bisulfate anions and solvate water molecules all linked by an extensive hydrogen-bonding network. Each Ru ion is coordinated in a distorted octahedral fashion by five ammonia molecules and a chloride ion. The Ru--Cl distance [2.3742 (5) A] is typical for RuIII--Cl linkages. The Ru--NH3 distances span a narrow range [2.096 (2)-2.119 (2) A] and provide no evidence for a trans effect.

Molecular Structure↗
PubMed2006

Potentiometric behaviour of ion selective electrodes based on iron porphyrins: the influence of porphyrin substituents on the response properties and analytical determination of diclofenac in pharmaceutical formulations.

The potentiometric response characteristics of diclofenac selective electrodes based on Fe(III) tetraphenylporphyrin-chloride (Fe(III)TPP-Cl) and Fe(III) tetrakis(pentafluorophenyl)porphyrin-chloride (Fe(III)TPFPP-Cl) in different mediator solvents and ionic additives are compared. The sensitivity, working range, detection limit, response mechanism, and selectivity of the membrane sensor show a significant dependence on the type of carrier substituent and on the pH value of the sample solution. Studies performed with different amounts of cationic additive (tetra-n-octylammoniumbromide (TOABr)) and anionic additive (sodium tetraphenylborate (NaTPB)) in the membranes allowed the determination of the potentiometric mechanism of action of the used metalloporphyrins. For the analysis of real samples, Fe(III)TPFPP-Cl (type G), prepared in o-NPOE, incorporating 10 mol% of TOABr, was used. This potentiometric unit presented a linear response towards diclofenac concentrations between 10(-5) and 10(-2)mol l(-1) ( I=0.1 mol l(-1)) and slopes of about -59 mV dec(-1), exhibiting a response time of 10s in a buffered solution of ammonia-ammonium sulphate with pH 9.9. The potentiometric analysis of sodium diclofenac in pharmaceutical formulations was carried out by direct potentiometry and the obtained results were compared to those provided by HPLC, presenting relative errors inferior to 1.0%.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed2002

The global distribution of acidifying wet deposition.

The acid-base status of precipitation is a result of a balance between acidifying compounds--mainly oxides of sulfur and nitrogen--and alkaline compounds--mainly ammonia and alkaline material in windblown soil dust. We use current models of the global atmospheric distribution of such compounds to estimate the geographical distribution of pH in precipitation and of the rate of deposition of hydrogen ion or bicarbonate ion. The lowest pH values--mainly due to high concentration of sulfuric acid--occur in eastern parts of North America, Europe, and China. A comparison with observed pH values shows fair agreement in most parts of the world. However, in some areas, e.g. western North America, southwestern Europe, and northern China the estimated pH is too low, indicating that we have underestimated the deposition flux of alkaline material, probably mainly CaCO3. Our neglect of organic acids may have contributed to an overestimate of pH especially in certain tropical areas. To illustrate the potential effects of acidifying deposition on nitrogen saturated terrestrial ecosystems we also calculate the deposition of "potential acidity" that takes into account the microbial transformation of ammonium to nitrate in such ecosystems, resulting in the release of hydrogen ion. Compared to the deposition of acidity, with its maxima over Europe, eastern North America, and southern China, the deposition of potential acidity exhibits an additional maximum in India and Bangladesh and in several other smaller hot spots where the cycling of ammonia is enhanced by a dense cattle population. To the extent that soils in these areas of high potential acidity deposition actually become nitrogen saturated a depletion of base cations and other changes in soil chemistry and biology should be expected. Potential problem areas forfuture soil acidification include several regions with sensitive soils in southern, southeastern, and eastern Asia as well as in central parts of South America.

Acid Rain↗
PubMed2006

Ammonium toxicity and potassium limitation in yeast.

DNA microarray analysis of gene expression in steady-state chemostat cultures limited for potassium revealed a surprising connection between potassium and ammonium: potassium limits growth only when ammonium is the nitrogen source. Under potassium limitation, ammonium appears to be toxic for Saccharomyces cerevisiae. This ammonium toxicity, which appears to occur by leakage of ammonium through potassium channels, is recapitulated under high-potassium conditions by over-expression of ammonium transporters. Although ammonium toxicity is well established in metazoans, it has never been reported for yeast. To characterize the response to ammonium toxicity, we examined the filtrates of these cultures for compounds whose excretion might serve to detoxify the ammonium (such as urea in mammals). Using liquid chromatography-tandem mass spectrometry to assay for a wide array of metabolites, we detected excreted amino acids. The amounts of amino acids excreted increased in relation to the severity of growth impairment by ammonium, suggesting that amino acid excretion is used by yeast for ammonium detoxification.

Amino Acids↗