Interactions between N application rate, CH4 oxidation and N2O production in soil

Here we report on a controlled environment experiment in which we applied ¹³C- and ¹⁵N-enrichment approaches to quantify methane oxidation rates and source partition N₂O production in a silt loam soil following application of NH₄NO₃, enabling us to look for potential interactions between methane oxidation and nitrifier-N₂O production. ¹⁵N-N₂O, ¹⁴⁺¹⁵N-N₂O and CO₂ fluxes and mineral N concentrations were measured over a 23-day period after application of NH₄NO₃ (5 at.% excess ¹⁵N) at rates of 0, 5, 10, 20, 30 and 40 g N m^?2 to a silt loam soil. Change in ^12/13C-CH₄ concentrations (as indicative of ¹³C-CH₄ oxidation rates) and production of ¹³C-CO₂ were monitored over the first 72 h after addition of 1.7 ??l ¹³C-CH₄ l^?1 (10 at.% excess ¹³C) to these N treatments. Oxidation of applied ¹³C-CH₄ was slower in the 5, 10, 20 and 30 g N m^?2 (5 at.% excess ¹⁵N) treatments (0.24?C0.32 ??g ¹³C-CH₄ l^?1 day^?1) than in the control (0.40 ??g ¹³C-CH₄ l^?1 day^?1), suggesting that these N loadings inhibited oxidation. N₂O production was raised after N addition, and in the 10, 20 and 30 g N m^?2 treatments nitrification was the predominant source of N₂O accounting for 61, 83 and 57% of the total ¹⁵N-N₂O produced, respectively. Our results point towards the possibility of methylotrophs switching function to oxidise ammonia in the presence of N, which may result in greater atmospheric loading of both CH₄ and N₂O.     

Rate of accumulation and emission of N2, N2O and CH4 from a flooded rice soil

In a field experiment using microplots, a flooded Crowley silt loam (Typic Albaqualfs) rice soil was fertilized with ¹⁵N labelled (60–74 atom %) urea and KNO₃. Emission of N₂, N₂O and CH₄ and accumulation in soil were measured for 21 d after fertilizer application.Emission of ¹⁵N₂-N measured from the urea and KNO₃ treated plots ranged from <15 to 570 and from 330 to 3,420 g ha^–1 d^–1, respectively. Entrapped ¹⁵N₂-N in the urea treated microplots was significantly lower (<15 g to 2.1 kg ha^–1) on all sampling dates compared to the ¹⁵N₂-N gas accumulation in the KNO₃ treated plots (6.4 to 31.5 kg ha^–1). Emissions of N₂O-N were low and did not exceed 4 g ha^–1 d^–1. Fluxes of CH₄ from the fertilizer and control plots were low and never exceeded 33 g ha^–1 d^–1. Maximum accumulation of CH₄ in the flooded soil measured 460 and 195 g ha^–1 for the urea and KNO₃ treatments, respectively.     

Design,synthesis and biological evaluation of N2,N4-disubstituted-1,1,3-trioxo-2H,4H-pyrrolo[1,2-b][1,2,4,6]thiatriazine derivatives as HIV-1 NNRTIs

A series of N₂,N₄-disubstituted-1,1,3-trioxo-2H,4H-pyrrolo[1,2-b][1,2,4,6]thiatriazine derivatives (PTTDs) was designed and synthesized by a facile route. The biological assay results showed that five most potent compounds displayed inhibitory activity against HIV-1 at low micromolar concentrations (EC₅₀ = 5.1–8.9 μM). Structure–activity relationship analysis indicated that N₂-(3-halogenated-benzyl) analogues were more potent than N₂-(unsubstituted-benzyl) analogues. The N₄-substitutions contributed to the antiviral activity in the following order: 2-/3-cyano substituted benzyl > 2-/3-halogenated benzyl > non-substituted benzyl > 4-halogenated benzyl. Docking studies of the representative compound revealed the binding conformation of these compounds and provided critical insights for the further development of PTTD analogues.     

Synthesis of N2, N2, 7-trimethylguanosine cap derivatives.

Several derivatives of N2,N2-7-trimethylguanosine (m3(2,2,7G)-cap, which was found at the 5' ends of small nuclear RNAs, were synthesized by use of S-phenyl N2,N2,7-trimethylguanosine 5'-phosphorothioate (PhSpm3(2,2,7)G) as a key intermediate. This compound was activated by iodine in the presence of phosphoric acid and diphosphoric acid to give N2,N2,7-trimethylguanosine-5'-diphosphate (ppm3(2,2,7)G) and 5'-triphosphate (ppm3(2,2,7)G), respectively. Similar reactions of PhSpm3(2,2,7)G with ADP and GDP gave capped dinucleoside triphosphates, m3(2,2,7)G5'pppA and m3(2,2,7)G5'pppG, respectively.     

Diurnal Oscillations in Gas Production (O2, CO2, CH4, and N2) in Soil Monoliths

Soil cores (35 cm long, 7 cm diameter) from the Macaulay Land Use Research Institute's Sourhope Research Station in the Scottish Borders were kept and monitored at constant temperature (18± 1°C) for gas production using a 1.6 mm diameter stainless steel probe fitted with a membrane inlet and connected to a quadrupole mass spectrometer. This provided a novel method for on-line, real time monitoring of soil gas dynamics. In closed-system headspace experiments, O₂ and CO₂ (measured at m/z values 32 and 44, respectively) showed anti-phase diurnal fluctuations in low-intensity simulated daylight and under a light-dark (LD, 12:12 h) regime. O₂ increased during periods of illumination and decreased in the dark. The inverse was true for CO₂ production. Ar (m/z = 40) concentration and temperature (°C) remained constant throughout the experiments. The same phase-related oscillations, in CO₂ and O₂ concentrations, were observed at 2 and 5 cm depth in soil cores. The O₂ concentration did not oscillate diurnally at 10 cm depth. In below-ground experiments, CH₄ (m/z = 15) concentration showed diurnal cycles at 2, 5 and 10 cm depth. The CH₄ production had the same diurnal phase cycle as CO₂ but with lower amplitude. Evidence of below-ground diurnal oscillations in N₂ (m/z = 28) concentration was provided at 5 cm depth. The scale of production and consumption of gases associated with soil-atmosphere interactions and below-ground processes, are shown to be a multifaceted output of several variables. These include light, circadian-controlled physiological rhythms of plants and microbes, and the interactions between these organisms.     

Diurnal Oscillations in Gas Production (O2, CO2, CH4, and N2) in Soil Monoliths

Soil cores (35 cm long, 7 cm diameter) from the Macaulay Land Use Research Institute's Sourhope Research Station in the Scottish Borders were kept and monitored at constant temperature (18± 1°C) for gas production using a 1.6 mm diameter stainless steel probe fitted with a membrane inlet and connected to a quadrupole mass spectrometer. This provided a novel method for on-line, real time monitoring of soil gas dynamics. In closed-system headspace experiments, O₂ and CO₂ (measured at m/z values 32 and 44, respectively) showed anti-phase diurnal fluctuations in low-intensity simulated daylight and under a light-dark (LD, 12:12 h) regime. O₂ increased during periods of illumination and decreased in the dark. The inverse was true for CO₂ production. Ar (m/z = 40) concentration and temperature (°C) remained constant throughout the experiments. The same phase-related oscillations, in CO₂ and O₂ concentrations, were observed at 2 and 5 cm depth in soil cores. The O₂ concentration did not oscillate diurnally at 10 cm depth. In below-ground experiments, CH₄ (m/z = 15) concentration showed diurnal cycles at 2, 5 and 10 cm depth. The CH₄ production had the same diurnal phase cycle as CO₂ but with lower amplitude. Evidence of below-ground diurnal oscillations in N₂ (m/z = 28) concentration was provided at 5 cm depth. The scale of production and consumption of gases associated with soil-atmosphere interactions and below-ground processes, are shown to be a multifaceted output of several variables. These include light, circadian-controlled physiological rhythms of plants and microbes, and the interactions between these organisms.     

Fluxes of N2O,CH4 and CO2 on afforested boreal agricultural soils

After drainage of natural boreal peatlands, the decomposition of organic matter increases and peat soil may turn into a net source of CO₂ and N₂O, whereas CH₄ emission is known to decrease. Afforestation is a potential mitigation strategy to reduce greenhouse gas emission from organic agricultural soils. A static chamber technique was used to evaluate the fluxes of CH₄, N₂O and CO₂ from three boreal organic agricultural soils in western Finland, afforested 1, 6 or 23 years before this study. The mean emissions of CH₄ and N₂O during the growing seasons did not correlate with the age of the tree stand. All sites were sources of N₂O. The highest daily N₂O emission during the growing season, measured in the oldest site, was as high as 29 mg N₂O m^–2d^–1. In general, organic agricultural soils are sinks for methane. Here, the oldest site acted as a small sink for methane, whereas the two youngest afforested organic soils were sources for methane with maximum emission rates (up to 154 mg m^–2d^–1) similar to those reported for minerogenous natural peatlands. Soil respiration rates decreased with the age of the forest. The high soil respiration in the younger sites, probably resulted from the high biomass production of herbs, could create soil anaerobiosis and increase methane production. Our results show that afforestation of agricultural peat soils does not abruptly terminate the N₂O emissions during the first two decades, and afforestation can even enhance methane emission for a few years. The carbon accumulation in the developing tree stand can partly compensate the carbon loss from soil.     

Predominance of the 1,N2-propano 2'-deoxyguanosine adduct among 4-hydroxy-2-nonenal-induced DNA lesions

4-Hydroxy-2-nonenal (HNE), one of the main aldehydic compounds released during lipid peroxidation, has been proposed to react with DNA bases in cells. Several classes of DNA lesions involving addition of either HNE or its 2,3-epoxide (epox-HNE) have been identified. In the present work, HPLC associated with tandem mass spectrometry was used to determine the pattern of HNE-induced DNA lesions. First, adducts were quantified within isolated DNA treated with HNE under peroxidizing conditions. The 1,N2-propano-2'-deoxyguanosine adduct of HNE (HNE-dGuo) was found to be the major lesion under all conditions studied. 1,N6-Ethenoadenine and 1,N2-ethenoguanine together with their (1,2-dihydroxyheptyl)-substituted derivatives, which all arise from the reaction of epox-HNE with DNA, were produced in significantly lower yields, even in the presence of 20 mM H2O2. The pyrimidopurinone malondialdehyde-2'-deoxyguanosine adduct was also found to be produced, although in very low yield. Similar results were obtained in cultured human monocytes incubated with HNE, because the HNE-dGuo adduct represented more than 95% of the overall adducts to DNA. In addition, the former lesion was poorly repaired, in contrast to 1,N2-ethenoguanine and, to a lesser extent, 1,N6-ethenoadenine. Altogether, these results suggest than HNE-dGuo may represent the best biomarker of the genotoxic effects of HNE.     

Analysis of ifosfamide, 4-hydroxyifosfamide, N2-dechloroethylifosfamide, N3-dechloroethylifosfamide and iphosphoramide mustard in plasma by gas chromatography-mass spectrometry

A sensitive and specific method for the simultaneous quantitation of ifosfamide (IF), 4-hydroxylifosfamide (4-OHIF), N2-dechloroethylifosfamide (N2D), N3-dechloroethylifosfamide (N3D) and iphosphoramide mustard (IPM) has been developed using gas chromatography-mass spectrometry (GC-MS) with an ion-trap mass spectrometer. Deuterium labeled analogues for each of these analytes were synthesized as the internal standards. The labile 4-OHIF in plasma was first converted to the more stable cyanohydrin adducts before dichloromethane extraction. IPM was extracted by C₁₈ reversed-phase resin. All analytes were converted to their silyl derivatives before GC-MS analysis. The sensitivity limits ranged from 0.1 to 0.5 μg/ml when 100 μl of plasma was used. This method was validated with within-run coefficients of variation less than 5% (n=8) and between-run coefficients of variation less than 12% (n=6). The method was applied to the determination of plasma levels of IF and metabolites in the rat.     

Synthesis of 3, N4-Dimethylcytidine

Abstract

Two procedures are described for the synthesis of 3, N⁴-dimethylcytidine, a compound usually obtained in only trace quantities.     

Improving anti-trypanosomal activity of 3-aminoquinoxaline-2-carbonitrile N1,N4-dioxide derivatives by complexation with vanadium

New vanadium complexes of the type [V(IV)O(L)(2)], where L are 3-aminoquinoxaline-2-carbonitrile N(1),N(4)-dioxide derivatives, were prepared as an effort to obtain new anti-trypanosomal agents improving the bioactivity of the free ligands. Complexation to vanadium of the quinoxaline ligands leads to excellent antiprotozoal activity, similar to that of the reference drugs nifurtimox and benznidazole and in all cases higher than that of the corresponding free ligands. In addition, it is for the first time that the V((IV))O-quinoxaline complexes are reported as a family of anti-Trypanosoma cruzi agents. Finally, the anti-trypanosomal activity of these vanadium complexes could be explained on the basis of their lipophilicity and the electronic characteristics of the quinoxaline substituents.     

Biomass fast pyrolysis in a fluidized bed reactor under N2, CO2, CO, CH4 and H2 atmospheres

Biomass fast pyrolysis is one of the most promising technologies for biomass utilization. In order to increase its economic potential, pyrolysis gas is usually recycled to serve as carrier gas. In this study, biomass fast pyrolysis was carried out in a fluidized bed reactor using various main pyrolysis gas components, namely N₂, CO₂, CO, CH₄ and H₂, as carrier gases. The atmosphere effects on product yields and oil fraction compositions were investigated. Results show that CO atmosphere gave the lowest liquid yield (49.6%) compared to highest 58.7% obtained with CH₄. CO and H₂ atmospheres converted more oxygen into CO₂ and H₂O, respectively. GC/MS analysis of the liquid products shows that CO and CO₂ atmospheres produced less methoxy-containing compounds and more monofunctional phenols. The higher heating value of the obtained bio-oil under N₂ atmosphere is only 17.8 MJ/kg, while that under CO and H₂ atmospheres increased to 23.7 and 24.4 MJ/kg, respectively.     

Preparation of N 2, N 2'7-Trimethylguanosine Affinity Columns

Abstract

2,2,7-trimethylguanosine (TMG) binding proteins from human cells were purified through TMG-affinity columns. TMG synthesis was improved and the TMG obtained was shown to be similar to the TMG in the 5′ cap of the UsnRNAs. The eluates obtained with TMG-affinity chromatographies were very different from those isolated with m⁷G-affinity columns, thus suggesting that specific TMG-binding proteins were obtained. The fraction may be enriched with factors associated with import and/or hypermethylation of UsnRNPs.     

Determination of N6-Threoninocarbonyladenosine,N2, N2-Dimethylguanosine,Pseudouridine and Other Ribonucleosides in Human Breast Milk

Abstract

Seven modified ribonucleosides from degraded tRNA and rRNA were quantified in milk from mothers with preterm infants. The amounts of N⁶-threoninocarbonyladenosine, N², N²-dimethylguanosine and pseudouridine supplied in the milk have been estimated and related to the respective urinary amounts excreted by preterm infants.     

Effects of CO2, CH4 and N2 on growth and nutrition of rice seedlings

Summary Flooded soils, which accumulate gaseous products of anaerobic fermentation, are often associated with poor rice plant growth. In the present experiment the effects of CO₂, CH₄, N₂, and air on rice seedling growth and nutrition were evaluated. Nutrient culture techniques were used to avoid secondary soil effects normally experienced.Carbon dioxide gas in the root zone of rice reduced seedling growth significantly, whereas CH₄ and N₂ had no significant effect. Methane gave no stimulatory benefits, unlike results reported by some earlier workers. Of three major nutrient elements studied, P uptake was affected more than N or K. Phosphorus uptake was significantly reduced in leaves and sheaths by all three gases, but was significantly increased in roots. This suggests an immobilization mechanism affecting P in roots, and since CO₂, CH₄, and N₂ behaved similarly in contrast to air, a lack of oxygen in the root system is suspected as the causal mechanism rather than toxic effects of gases. Effects on N and K uptake were minimal and insignificant.Contribution from the Department of Agronomy and Range Science, University of California, Davis, California 95616.Contribution from the Department of Agronomy and Range Science, University of California, Davis, California 95616.     

Syntheses of optically active 2-amino-4-oxobutyric acid and N,O-protected derivatives

Summary Strategies for the synthesis of optically active aspartaldehyde derivatives are reviewed. Most of them are using the chiral pool: allylglycine or naturally occurring homoserine, aspartic acid or methionme and side chain modifications. This will be developed in the first part. Some other original routes are also displayed in the second part. Different aspects of each strategy are discussed: the nature and number of steps, the problem of protecting groups, the price and availability of starting materials. Some synthetic applications of such interesting chiral synthons are shown in the last part.Abbreviations Ac acetyl - An Anisyl or 4-methoxy benzyl - Bn benzyl - Boc tert-butoxycarbonyl - BOP-PF₆ benzotriazol-l-yloxytris(dimethylamino)phosphonium hexafluorophosphate - Cbz benzyloxycarbonyl - DCC dicyclohexylcarbodiimide - DIBAL diisobutyl aluminum hydride - DIPEA diisopropyl ethyl amine - DMF dimethyl formamide - DMSO dimethylsulfoxide - EDCI l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride - HP 4-hydroxy phenyl - MP 4-methoxy phenyl - NCS N-chlorosuccinimide - NMR nuclear magnetic resonance - PCC pyridinium chlorochromate - Pht phthaloyl - Ser serine - tBu tert-butyl - TEMPO 2,2,6,6-tetramethyl piperidine-l-oxyl - TFA trifluoro acetic acid - Trityl triphenyl methyl - Val valine This paper is dedicated to RV.     

N1,N5,N10-Tris(4-hydroxycinnamoyl)spermidines from Microdesmis keayana roots

Three new N1,N5,N10-tris(4-hydroxycinnamoyl)spermidines were isolated from a methanolic root extract of Microdesmis keayana. They were identified as N5,N10-di(p-coumaroyl)-N1-feruloylspermidine,N5-(p-coumaroyl)-N1,N10-diferuloylspermidine, and N1,N5,N10-triferuloylspermidine, and were named keayanidines A, B, and C (1-3), respectively. Their structures were established by spectral techniques(electrospray mass spectrometry, one- and two-dimensional NMR). A 4',4',4'-trimethylated derivative was prepared by methylation of keayanidine C, and the same compound was synthesized fromspermidine and 3,4-dimethoxycinnamic acid to confirm the spectral attributions of the NMR data of the natural compounds. Radical-scavenging properties of all compounds were evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical spectrophotometric assay.