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At least 919 records · Page 51Linked to original sources
PubMed2000

Synthesis, conformational analysis, and biological activity of C-thioribonucleosides related to tiazofurin.

The syntheses of furanthiofurin [5beta-D-(4'-thioribofuranosyl)furan-3-carboxamide, 1] and thiophenthiofurin [5beta-D-(4'-thioribofuranosyl)thiophene-3-carboxamide, 2], two C-thioribonucleoside analogues of tiazofurin, are described. Direct trifluoroacetic acid-catalyzed C-glycosylation of ethyl furan-3-carboxylate with 1-O-acetyl-2,3,5-tri-O-benzyl-4-thio-D-ribofuranose gave 2- and 5-glycosylated regioisomers, as a mixture of alpha and beta anomers. Ethyl 5-(2,3,5-tri-O-benzyl)-beta-D-(4'-thioribofuranosyl)furan-3-carboxylate (6beta) was debenzylated and then converted into the corresponding amide (furanthiofurin) by reaction with ammonium hydroxide. A similar C-glycosylation of ethyl thiophene-3-carboxylate with 1,2,3,5-tetra-O-acetyl-4-thio-D-ribofuranose catalyzed by stannic chloride afforded an anomeric mixture of 2- and 5-glycosylated regioisomers. Deacetylation of ethyl 5-(2,3,5-tri-O-acetyl)-beta-D-(4'-thioribofuranosyl)thiophene-3-carboxylate (13beta) with methanolic ammonia and treatment of the ethyl ester with ammonium hydroxide gave thiophenthiofurin. The glycosylation site and anomeric configuration were established by (1)H NMR spectroscopy. Thiophenthiofurin was found to be cytotoxic in vitro toward human myelogenous leukemia K562, albeit 39-fold less than thiophenfurin, while furanthiofurin proved to be inactive. K562 cells incubated with thiophenthiofurin resulted in inhibition of inosine 5'-monophosphate dehydrogenase (IMPDH) and an increase in IMP pools with a concurrent decrease in GTP levels. From computational studies it was deduced that, among the C-nucleoside analogues of tiazofurin, activity requires an electrophilic sulfur adjacent to the C-glycosidic bond and an energetically favorable conformer around chi = 0 degrees. Among these, the more constrained (least flexible) compounds (tiazofurin and thiophenfurin) are more active than the less constrained thiophenthiofurin. Those compounds which contain a nucleophilic oxygen in place of the thiazole or thiophene (oxazofurin, furanfurin, and furanthiofurin) show the least activity.

Antineoplastic Agents↗
PubMed1993

Characterization of a nitrophenol reductase from the phototrophic bacterium Rhodobacter capsulatus E1F1.

The phototrophic bacterium Rhodobacter capsulatus E1F1 photoreduced 2,4-dinitrophenol to 2-amino-4-nitrophenol by a nitrophenol reductase activity which was induced in the presence of nitrophenols and was repressed in ammonium-grown cells. The enzyme was located in the cytosol, required NAD(P)H as an electron donor, and used several nitrophenol derivatives as alternative substrates. The nitrophenol reductase was purified to electrophoretic homogeneity by a simple method. The enzyme was composed of two 27-kDa subunits, was inhibited by metal chelators, mercurial compounds, and Cu2+, and contained flavin mononucleotide and possibly nonheme iron as prosthetic groups. Purified enzyme also exhibited NAD(P)H diaphorase activity which used tetrazolium salt as an electron acceptor.

2,4-Dinitrophenol↗
PubMedDate not supplied

[Mechanism of cyanide and thiocyanate decomposition by an association of Pseudomonas putida and Pseudomonas stutzeri strains].

The intermediate and terminal products of cyanide and thiocyanate decomposition by individual strains of the genus Pseudomonas, P. putida strain 21 and P. stutzeri strain 18, and by their association were analyzed. The activity of the enzymes of nitrogen and sulfur metabolism in these strains was compared with that of the collection strains P. putida VKM B-2187T and P. stutzeri VKM B-975T. Upon the introduction of CN- and SCN- into cell suspensions of strains 18 and 21 in phosphate buffer (pH 8.8), the production of NH4+ was observed. Due to the high rate of their utilization, NH3, NH4+, and CNO- were absent from the culture liquids of P. putida strain 21 and P. stutzeri strain 18 grown with CN- or SCN-. Both Pseudomonas strains decomposed SCN- via cyanate production. The cyanase activity was 0.75 micromol/(min mg protein) for P. putida strain 21 and 1.26 micromol/(min mg protein) for P. stutzeri strain 18. The cyanase activity was present in the cells grown with SCN- but absent in cells grown with NH4+. Strain 21 of P. putida was a more active CN- decomposer than strain 18 of P. stutzeri. Ammonium and CO2 were the terminal nitrogen and carbon products of CN- and SCN- decomposition. The terminal sulfur products of SCN- decomposition by P. stutzeri strain 18 and P. putida strain 21 were thiosulfate and tetrathionate, respectively. The strains utilized the toxic compounds in the anabolism only, as sources of nitrogen (CN- and SCN-) and sulfur (SCN-). The pathway of thiocyanate decomposition by the association of bacteria of the genus Pseudomonas is proposed based on the results obtained.

Ammonia↗
PubMed1985

[Measure of the activity of a quaternary aldehyde-ammonia complex against reference bacteria strains and 50 hospital strains].

Concentrations of a disinfectant complex effective against hospital strains were determined using the dilution-neutralization method (AFNOR T72 150). Results were compared to those obtained with reference bacterial strains. A 99.999% reduction in viable bacteria requires significantly higher concentrations for hospital isolates than for reference strains.

Acinetobacter↗
PubMed1976

[Changes in blood sugar and ammonium levels after ligation of the afferent hepatic vessels].

Blood sugar and ammonia levels were studied after ligation of the afferent vessels of the liver. Isolated liver anoxia and isolated splanchnic stagnation were produced by means of a side-to-side porto-caval anastomosis in dogs. Isolated liver ischaemia for 45 minutes did not lead to a typical change in blood sugar and ammonia levels. In splanchnic stagnation with intact hepatic circulation, hyperglycaemia and a high blood ammonia level appeared in the portal venous blood. Thus, in the first 45 minutes afferent hepatic vessels are primarily due to splanchnic stagnation and not to hepatic ischaemia. When splanchnic stagnation and hepatic ischaemia occur simulatneously, the enhanced production of metabolites and the lack of compensation lead to a rapid lethal outcome.

Animals↗
PubMedDate not supplied

[Utilization of nitrogen compounds by phototrophic bacteria].

Phototrophic purple and green bacteria differ by their ability to assimilate nitrogen compounds. Thiocapsa roseopersicina utilizes not only ammonium and nitrates, as a source of nitrogen, but also urea, azoguanine, cytosine and some amino acids. The non-sulphur bacterium Rhodopseudomonas palustris grows at the account of amine nitrogen of a larger number of amino acids than the sulphur bacterium. Chlorobium limicola 1. thiosulfatophilum grows only on media containing urea and methylamines. Formation of amino acids by the studied phototrophic bacterial cultures is related to amination of alpha-ketoglutarate. Purple bacteria possess also the activity of aspartate dehydrogenase.

Amino Acids↗
PubMed2007

Structural elucidation of 4-(cystein-S-yl)butyl glucosinolate from the leaves of Eruca sativa.

The structurally unique glucosinolate (GSL), 4-(cystein-S-yl)butyl GSL, was identified in the leaves of hydroponically-grown rocket salad (Eruca sativa Mill.). Its electrospray ionization mass spectrometry (ESI-MS)/MS spectrum indicated that this unusual GSL had a molecular weight of 414 as a desulfo (DS)-GSL, and a molecular formula of C(14)H(25)N(2)O(8)S(2) based on its negative ion matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) spectrum. For further confirmation, the 4-(cystein-S-yl)butyl DS-GSL was prepared with authentic L-Ser and purified dimeric 4-mercaptobutyl DS-GSL, and its chemical structure then confirmed by ESI-MS/MS data. It is named "glucorucolamine" as a trivial name from its ammonia sensitivity. This unique GSL was found to the greatest extent when rocket salad was grown in a 100% NH4+-N nutrient solution. Despite it clearly seems to reduce the detoxification of excess NH4+ in the leaves of rocket salad, present knowledge about the unique GSL is still far from being sufficient.

Brassicaceae↗
PubMed2004

[Possible role of ammonium as a paracrine regulator in the uterine tissue].

An opportunity of formation of ammonia (NH3) in utera endometrium and its influence on exchange of Ca2+ and H+ in plasmalemma of myometrium was investigated. Dissociation of endometrium stroma cells and myocytes suspension was carried from utera of pigs and rats in accordance with the traditional techniques. In suspension of stroma cells a rather high AMP-deaminase activity (53 +/- 2 mmol IMP/hour on 1 mg of protein) was determined. It was demonstrated that ammonia release in extracellular space (measured by the changes of colouring of trinitrobenzolsulfonate acid) was significantly amplified by 1 mM acetylcholine and decreased by 0,1 mM fluoride ions, nonspecific AMP-deaminase inhibitor. It enables to assume a role of AMP-deaminase in formation of NH3 by endometrium stroma cells and its release into extracellular space during acetylcholine stimulation. The addition of ammonia (4 mM) to suspension of myocytes is accompanied by significant increase in pH (measured by the change in BCECF fluorescence) in extracellular and intracellular space, and the last parameter is inhibited by the blockers of passive H+ transport across the membrane: 0,1 mM 4-aminopyridine and tetraethylammonium. It is possible that addition of ammonia-containing solution results in increase in proton gradient on myocyte membrane and in amplification of H+ efflux. The opportunity of stimulation ofacetylcholine-activated passive Ca2+ transport in myocytes by 4 mM NH4+ that was suppressed by 1 mM cadmium and 1 nM nifedipine was also shown using fluorescent probe FURA-2AM. The increase in Ca2+ concentration in cytoplasm in the given conditions is intensively oppressed by protonophore (0.04% 2,4-dinitrophenol) and is effectively amplified by Na+/H+-exchange inhibitor 0,1 mM amyloride. It is possible to assume an amplification of lygand-activated passive Ca2+ transport caused by dispersion of transmembrane proton gradient which exists on plasmalemma and can be increased by ammonia formation in endometrium. The role of diffused from endometrium NH3 in regulation of utera functional activity requires further investigation, however already at this stage it is possible to assume, that NH3 molecules (or ion NH4+) can carry out a role of paracrine regulator in the system endometrium-myometrium.

AMP Deaminase↗
PubMed2004

Ammonium and hydroxylamine uptake and accumulation in Nitrosomonas.

Starved cells of Nitrosomonas europaea and further ammonia oxidizers were able to rapidly accumulate ammonium and hydroxylamine to an internal concentration of about 1 and 0.8 M, respectively. In kinetic studies, the uptake/accumulation rates for ammonium [3.1 mmol (g protein)(-1) min(-1)] and hydroxylamine [4.39 mmol (g protein)(-1) min(-1)] were determined. The uptake and accumulation process of ammonium and hydroxylamine was not coupled to ammonia or hydroxylamine oxidation and nitrite was not produced. In the presence of uncouplers the ammonium accumulation was completely inhibited, indicating an active, membrane-potential-driven transport mechanism. When the external ammonium or hydroxylamine pool was depleted, the internal ammonium and hydroxylamine was consumed within 12 h or 20 min, respectively. The binding of ammonium/ammonia was correlated with an energized membrane system, and hydroxylamine may bind to the hydroxylamine oxidoredutase.

Culture Media↗
PubMed2000

Biomass production of the mycorrhizal fungus Suillus grevillei: effect of pH and ammonium.

The ability of the ectomycorrhizal fungus Suillus grevillei (Klotzsch) Singer to grow in agitated submerged culture was investigated by employing the Marx-Melin-Norkrans (MMN) medium. The operating conditions suitable for improving the biomass production were determined. Batch experimental tests were carried out in either shake flasks or a stirred tank reactor. The results showed that at least two factors strongly affected the fungal growth, namely the pH and the ammonia-nitrogen concentration in the medium. By controlling the acidity in the pH range 4-5 with a Na-citrate buffer solution and introducing the ammonia-nitrogen in a step-feed way (without exceeding a concentration of approximately 0.07 kg N/m3), the exponential growth phase continued for longer than that of the control culture (no stationary phase seemed to be reached after 17 days) and an approximately 2-fold increase of the biomass/substrate growth yield was obtained compared with the control culture.

Basidiomycota↗
PubMed1991

Regulation of glutamine synthetase activity in the unicellular cyanobacterium Synechocystis sp. strain PCC 6803 by the nitrogen source: effect of ammonium.

Glutamine synthetase activity from Synechocystis sp. strain PCC 6803 is regulated as a function of the nitrogen source available in the medium. Addition of 0.25 mM NH4Cl to nitrate-grown cells promotes a clear short-term inactivation of glutamine synthetase, whose enzyme activity decreases to 5 to 10% of the initial value in 25 min. The intracellular levels of glutamine, determined under various conditions, taken together with the results obtained with azaserine (an inhibitor of transamidases), rule out the possibility that glutamine per se is responsible for glutamine synthetase inactivation. Nitrogen starvation attenuates the ammonium-mediated glutamine synthetase inactivation, indicating that glutamine synthetase regulation is modulated through the internal balance between carbon-nitrogen compounds and carbon compounds. The parallelism observed between the glutamine synthetase activity and the internal concentration of alpha-ketoglutarate suggests that this metabolite could play a role as a positive effector of glutamine synthetase activity in Synechocystis sp. Despite the similarities of this physiological system to that described for enterobacteria, the lack of in vivo 32P labeling of glutamine synthetase during the inactivation process excludes the existence of an adenylylation-deadenylylation system in this cyanobacterium.

Ammonia↗
PubMed1971

Studies on the porphobilinogen deaminase-uroporphyrinogen cosynthetase system of cultured soya-bean cells.

1. Porphobilinogenase was isolated and purified from soya-bean callus tissue; its components, porphobilinogen deaminase and uroporphyrinogen isomerase, were separated and purified. 2. The purified porphobilinogenase was resolved into two bands on starch-gel electrophoresis. The molecular weights of porphobilinogenase, deaminase and isomerase fractions were determined by the gel-filtration method. Porphobilinogenase activity was affected by the presence of air; uroporphyrinogens were only formed under anaerobic conditions, although substrate consumption was the same in the absence of oxygen as in its presence. 3. pH-dependence of both porphobilinogenase and deaminase was the same and a sharp optimum at pH 7.2 was obtained. Isomerase was heat-labile, but the presence of ammonium ions or porphobilinogen afforded some protection against inactivation. The action of several compounds added to the system was studied. Cysteine, thioglycollate, ammonium ions and hydroxylamine inhibited porphobilinogenase; certain concentrations of sodium and magnesium salts enhanced activity; some dicarboxylic acids and 2-methoxy-5-nitrotropone inhibited the deaminase. 4. delta-Aminolaevulate and ethionine in the culture media stimulated porphyrin synthesis and increased porphobilinogenase activity, whereas iron deficiency resulted in porphyrin accumulation. 5. The development of chlorophyll and porphobilinogenase on illumination of dark-grown callus was followed. 6. A hypothetical scheme is suggested for the enzymic synthesis of uroporphyrinogens from porphobilinogen.

Amino Acids↗
PubMed2002

Oxygen-limited autotrophic nitrification/denitrification by ammonia oxidisers enables upward motion towards more favourable conditions.

The hypothesis is formulated that in case of oxygen limitation in the sediment, nitrifiers switch from nitrification to oxygen-limited autotrophic nitrification-denitrification (OLAND) in order to survive and maintain activity. During OLAND, ammonium is oxidised using nitrite as e-acceptor to form dinitrogen gas. As an additional advantage they benefit from the gaseous N(2) formed as a means of transport. In this way, the nitrifiers can move out of the sediment and rise through the water column towards more favourable conditions. At the surface, the bacteria could take up oxygen, and recommence nitrification. In order to test this hypothesis, nitrifying sediment with an overlaying water column was simulated in lab-scale columns. Nitrogen transformations and material transport through the water column were followed after addition of different forms of nitrogen under oxygen-limited conditions. (15)N-labelling experiments showed a large contribution of OLAND to the observed nitrogen deficits. Nitrifier enumerations, fluorescent in situ hybridisation and 16S rRNA gene analysis revealed increased populations of ammonia oxidising nitrifiers in the upper water layers. The results presented support the proposed hypothesis of transport using OLAND. Nitrifying activity in the sediment immediately recovered almost completely from prolonged oxygen-limited incubation when oxygen concentrations were increased.

Anaerobiosis↗
PubMed2005

Relevance of ammonium oxidation within biological soil crust communities.

Thin, vertically structured topsoil communities that become ecologically important in arid regions (biological soil crusts or BSCs) are responsible for much of the nitrogen inputs into pristine arid lands. We studied N(2) fixation and ammonium oxidation (AO) at subcentimetre resolution within BSCs from the Colorado Plateau. Pools of dissolved porewater nitrate/nitrite, ammonium and organic nitrogen in wetted BSCs were high in comparison with those typical of aridosoils. They remained stable during incubations, indicating that input and output processes were of similar magnitude. Areal N(2) fixation rates (6.5-48 micromol C(2)H(2) m(-2) h(-1)) were high, the vertical distribution of N(2) fixation peaking close to the surface if populations of heterocystous cyanobacteria were present, but in the subsurface if they were absent. Areal AO rates (19-46 micromol N m(-2) h(-1)) were commensurate with N(2) fixation inputs. When considering oxygen availability, AO activity invariably peaked 2-3 mm deep and was limited by oxygen (not ammonium) supply. Most probable number (MPN)-enumerated ammonia-oxidizing bacteria (6.7-7.9 x 10(3) cells g(-1) on average) clearly peaked at 2-3 mm depth. Thus, AO (hence nitrification) is a spatially restricted but important process in the nitrogen cycling of BSC, turning much of the biologically fixed nitrogen into oxidized forms, the fate of which remains to be determined.

Cyanobacteria↗
PubMed1999

Use of molecular and isotopic techniques to monitor the response of autotrophic ammonia-oxidizing populations of the beta subdivision of the class proteobacteria in arable soils to nitrogen fertilizer.

This study examined the effects of NH(4)NO(3) fertilizer on the size and activity of nitrifying, autotrophic, ammonia-oxidizing populations of the beta subdivision of the class Proteobacteria in arable soils. Plots under different long-term fertilizer regimes were sampled before and after NH(4)NO(3) additions, and the rates of nitrification were determined by (15)N isotopic pool dilution assays. Ammonia-oxidizing populations in the plots were quantified by competitive PCR assays based on the amoA and ribosomal 16S genes. Prior to fertilizer addition, ammonium concentrations and nitrification rates in the plots were comparatively low; ammonia-oxidizing populations were present at 10(4) to 10(5) gene copies g of soil(-1). Three days after the application of fertilizer, nitrification rates had risen considerably but the size of the ammonia-oxidizing population was unchanged. Six weeks after fertilizer treatment, ammonium concentrations and nitrification rates had fallen while the ammonia-oxidizing populations in plots receiving fertilizer had increased. The rapidity of the rise in nitrification rates observed after 3 days suggests that it results from phenotypic changes in the ammonia-oxidizing bacterial population. Associated increases in population sizes were only observed after 6 weeks and did not correlate directly with nitrifying activity. Phylogenetic analyses of PCR products from one of the plots revealed a population dominated by Nitrosospira-type organisms, similar to those prevalent in other soils.

Betaproteobacteria↗
PubMed2004

Basolateral ammonium transport by the mouse inner medullary collecting duct cell (mIMCD-3).

The renal collecting duct is the primary site for the ammonia secretion necessary for acid-base homeostasis. Recent studies have identified the presence of putative ammonia transporters in the collecting duct, but whether the collecting duct has transporter-mediated ammonia transport is unknown. The purpose of this study was to examine basolateral ammonia transport in the mouse collecting duct cell (mIMCD-3). To examine mIMCD-3 basolateral ammonia transport, we used cells grown to confluence on permeable support membranes and quantified basolateral uptake of the radiolabeled ammonia analog [14C]methylammonia ([14C]MA). mIMCD-3 cell basolateral MA transport exhibited both diffusive and transporter-mediated components. Transporter-mediated uptake exhibited a Km for MA of 4.6 +/- 0.2 mM, exceeded diffusive uptake at MA concentrations below 7.0 +/- 1.8 mM, and was competitively inhibited by ammonia with a Ki of 2.1 +/- 0.6 mM. Transporter-mediated uptake was not altered by inhibitors of Na+-K+-ATPase, Na+-K+-2Cl(-) cotransporter, K+ channels or KCC proteins, by excess potassium, by extracellular sodium or potassium removal or by varying membrane potential, suggesting the presence of a novel, electroneutral ammonia-MA transport mechanism. Increasing the outwardly directed transmembrane H+ gradient increased transport activity by increasing Vmax. Finally, mIMCD-3 cells express mRNA and protein for the putative ammonia transporter Rh B-glycoprotein (RhBG), and they exhibit basolateral RhBG immunoreactivity. We conclude that mIMCD-3 cells express a basolateral electroneutral NH4+/H+ exchange activity that may be mediated by RhBG.

Acid-Base Equilibrium↗
PubMed1990

Stimulation of ammonification and nitrification in soils by the insecticides monocrotophos and quinalphos.

The application (up to 5 kg ha-1) of monocrotophos or quinalphos to four types of agricultural soils significantly stimulated the mineralization of peptone-nitrogen and the oxidation of ammonium-nitrogen. In soils treated with 2.5 kg ha-1 of either insecticide, the rate of ammonification and nitrification was fairly rapid after 2 and 4 weeks of incubation. The enhancement of both transformations, mediated by microorganisms, was significantly more in the quinalphostreated soils. The results suggest that a balance between the effect of insecticides on insect pests and their impact on beneficial microbial activities in soil must be determined.

Ammonia↗
PubMed1990

[Determination of traces of gallium in air by oscillographic polarography].

Gallium and its compounds in air collected by millipore filter membrane are leached by 10% HNO3 at room temperature. After being adjusted with ammonia solution to pH8-9, the leachate is filtered and the filtrate is evaporated to dryness on a water bath. The residue is dissolved with 2 ml of 10% HNO3 and then into the solution are added 0.5 ml of 5% hydroxylamine hydrochloride solution, 0.20 ml of 5% sulfosalicyclic acid solution, 0.5 ml of NaF solution (1mg F/ml) and 1 drop of methyl red solution. Let it stand for 5 min after the color of the solution turns orange by addition of 1:1 NH4OH solution. Add 0.20ml of 0.20% cupferron solution, 0.20ml of 0.5% diphenyl-guanidine solution (prepared in 95% ethanol) and 0.5ml of acetic acid-ammonium acetate buffer (pH 7.0). Then add water to make the final volume of the test solution to be 10ml and measure the height of the second derivative polarographic wave at -1.08V (vs.SCE). The sensitivity of the method is 0.005-0.10 microgram/ml, the coefficient of variation 6.8%, recovery 97.8%. Within 0.005-0.10 microgram/ml, the wave height is linear with the concentration of gallium.

Air↗