Oxidation of lipids. XI. Spin trapping and identification of peroxy and alkoxy radicals of methyl linoleate

Spin trapping of peroxy and alkoxy radicals generated from the hydroperoxide of methyl linoleate was studied using methyl-N-duryl nitrone (MDN) and phenyl-N-tert-butyl nitrone (PBN) as spin traps. The conjugated dienyl carbon radical was also generated from methyl linoleate and spin-trapped. The spin adducts of peroxy, alkoxy, and dienyl carbon radicals were observed by ESR and their hyperfine splitting constants were determined. The spin adducts of peroxy and alkoxy radicals could be distinguished clearly with MDN.     

Night and day: short‐term variation in nitrogen chemistry and nitrous oxide emissions from streams

1. Diel variation in metabolism contributes to variation in oxygen (O₂) concentrations in streams. This variation in O₂ and other parameters (e.g. pH) can in turn affect the rates of microbial nitrogen (N) processing, concentrations of nitrogenous solutes and production of the greenhouse gas nitrous oxide (N₂O). We investigated diel variability in emissions of N₂O and the magnitude of short‐term variability in N solutes across 10 streams. 2. Nitrous oxide fluxes varied on average 2.3‐fold over diel cycles. Concentrations would be underestimated by sampling around noon, but N₂O fluxes would not show a consistent bias. Time‐weighted mean daily N₂O flux was strongly related to nitrate concentration (r² = 0.58). Diel patterns in N₂O and dissolved N species were often complex (rather than simple sinusoidal curves), probably reflecting complex underlying processes. 3. Reliance on samples obtained around noon would overestimate daily mean nitrate concentrations by 5% and underestimate ammonium by 32% (average bias across all streams and dates). 4. Dissolved organic N did not show consistent day–night variation. However, the magnitude of diel variability was similar to that observed for dissolved inorganic N. Organic and inorganic N concentrations were often similar. Both appear to be dynamic components of stream N budgets. 5. The Intergovernmental Panel on Climate Change (IPCC) relies upon an emission factor to estimate indirect agricultural N₂O emissions from streams and ground water. The measured emission factor (defined as the ratio of concentrations of N₂O‐N to ‐N) was typically below the recently revised IPCC default figure. Measured values varied on average 1.8‐fold over approximately 24‐h periods and were slightly higher at night than by day. The emission factor was actually highest in streams that were net sinks for N₂O, highlighting a conceptual problem in the current IPCC method. 6. Typical sampling programmes rely on daytime‐only sampling, which might cause bias in results. In our study streams, the bias was generally small. Diel variation in nitrate concentrations was related to mean temperature; variation in ammonium and N₂O concentrations was greatest at low concentrations of nitrite and ammonium.     

Microbial removal of nitrogen monoxide (NO) under aerobic conditions

Nitrogen oxide gas (NO_x), consisting of nitrogen monoxide (NO) and nitrogen dioxide (NO₂), at a low concentration corresponding to that on roads as a result of exhaust from automobiles, was supplied for 25 days through a laboratory-scale biofilter packed with soil as a packing material. The removal efficiency of NO₂ by soil was almost 100%, and the removal efficiency of NO was 60% on average and 86% at maximum. By using -irradiated soil as a packing material, NO₂ was completely removed mainly by adsorption onto or absorption into the packing material. However, the removal efficiency of NO in the sterilized soil was only 20%, suggesting that NO in soil was removed microbiologically under aerobic conditions.     

Annual nitrous oxide flux and soil nitrogen characteristics in sagebrush steppe ecosystems

Soil nitrogen transformations and nitrous oxide fluxes were measured in a range of sagebrush steppe ecosystems in south-central Wyoming. Net nitrate production, measured in laboratory incubations, was highest in the ecosystem type dominated by Artemisia tridentata ssp. vaseyana, especially early in the growing season. Fluxes of nitrous oxide, measured in closed chambers and analyzed by gas chromatography, also tended to be higher in the same type, but only for short periods in the spring. Thereafter, all nitrous oxide fluxes were low and did not differ consistently among types. Estimated average annual fluxes for three Artemisia ecosystem types (dominated by Artemisia tridentata ssp. vaseyana, Artemisia tridentata ssp. wyomingensis, and Artemisia nova) were 0.32, 0.23 and 0.13 kg N₂O-N ha^–1 y^–1 repsectively. Average annual flux, weighted by the areal extent of these and other vegetation types in the region, was approximately 0.21 kg N₂O-N ha^–1y^–1. Assuming this landscape is representative of sagebrush steppe, we calculate a flux of 9.5 × 10⁹ g y^–1 of N₂O-N from U.S. sagebrush steppe, and a flux of 1.1 × 10¹¹ g y^–1 of N₂0-N from analogous desert and semi-desert shrublands of the world.     

HNO_2诱变选育链霉菌702菌株

以链霉菌702-20为出发菌株,经HNO2诱变处理,获得高产突变株。实验结果表明:HNO2处理20 m in对菌株的致死率可达83.10%,突变率高达14.13%,经过摇瓶筛选获得高产突变株20-29-148,产链霉素能力达到1.404 mg/mL,比出发菌株提高了37.65%。经传代培养考察,该突变菌株具有良好的遗传稳定性。     

Snow depth, soil freezing, and fluxes of carbon dioxide, nitrous oxide and methane in a northern hardwood forest

Soil–atmosphere fluxes of trace gases (especially nitrous oxide (N₂O)) can be significant during winter and at snowmelt. We investigated the effects of decreases in snow cover on soil freezing and trace gas fluxes at the Hubbard Brook Experimental Forest, a northern hardwood forest in New Hampshire, USA. We manipulated snow depth by shoveling to induce soil freezing, and measured fluxes of N₂O, methane (CH₄) and carbon dioxide (CO₂) in field chambers monthly (bi-weekly at snowmelt) in stands dominated by sugar maple or yellow birch. The snow manipulation and measurements were carried out in two winters (1997/1998 and 1998/1999) and measurements continued through 2000. Fluxes of CO₂ and CH₄ showed a strong seasonal pattern, with low rates in winter, but N₂O fluxes did not show strong seasonal variation. The snow manipulation induced soil freezing, increased N₂O flux and decreased CH₄ uptake in both treatment years, especially during winter. Annual N₂O fluxes in sugar maple treatment plots were 207 and 99 mg N m⁻² yr⁻¹ in 1998 and 1999 vs. 105 and 42 in reference plots. Tree species had no effect on N₂O or CO₂ fluxes, but CH₄ uptake was higher in plots dominated by yellow birch than in plots dominated by sugar maple. Our results suggest that winter fluxes of N₂O are important and that winter climate change that decreases snow cover will increase soil:atmosphere N₂O fluxes from northern hardwood forests.     

Spin trapping of glutathiyl and protein radicals produced from nitric oxide-derived oxidants

Despite the importance of protein radicals in cell homeostasis and cell injury, their formation, localization, and propagation reactions remain obscure, mainly because of the difficulties in detecting and characterizing radicals, in general, and protein radicals, in particular. New approaches based on spin trapping coupled with other methodologies are under development/testing but so far they have been applied mainly to the study of protein-tyrosyl and protein-tryptophanyl radicals. Here, our aim is to emphasize the importance of developing new methodologies for the detection of glutathyil and protein-cysteinyl radicals under physiological conditions. To this end, we summarize current EPR evidence supporting the view that glutathione and protein-cysteines are among the preferential targets of nitric oxide-derived oxidants and that they are oxidized to the glutathiyl and protein-cysteinyl radicals, respectively. The possible intermediacy of these species in the biological formation of mediators of protein-cysteine redox signaling, such as S-nitrosothiols and sulfenic acids, is also discussed.     

新疆天山高寒草原不同放牧管理下的CO_2,CH_4和N_2O通量特征

以中国科学院新疆巴音布鲁克草原生态站为依托,于2010年5月—2011年10月利用静态箱-气相色谱法对短期禁牧(2005年围封)、长期禁牧(1984年围封)和自由放牧(冬季放牧)3种草地的CO2、CH4、N2O气体通量进行了野外连续试验研究。结果表明:新疆天山高寒草原对CO2,CH4和N2O通量表现出明显的季节排放特点。在植物的生长季(5—10月),新疆天山高寒短期禁牧、长期禁牧和自由放牧草原的CO2通量平均值分别为:(89.8±49.3)、(52.8±28.7)、(57.0±30.7)mg·m-2·h-1,CH4通量平均值分别为:(-66.3±21.3)、(-104.5±32.8)、(-103.0±39.0)μg·m-2·h-1,N2O通量平均值分别为:(21.2±11.8)、(13.6±6.9)、(13.2±6.2)μg·m-2·h-1;短期禁牧草原与长期禁牧和自由放牧草原CH4平均通量具有显著性差异(P0.05),但CO2和N2O差异不显著(P0.05)。在植物的非生长季(11月—翌年4月),新疆天山高寒短期禁牧、长期禁牧以及自由放牧草原的3种温室气体的通量较低且差异均不显著。     

Spin trapping endogenous radicals in MC-1010 cells: Evidence for hydroxyl radical and carbon-centered ascorbyl radical adducts

Incubation of MC-1010 cells with the spin-trapping agent 5,5-dimethyl-1-pyrroline 1-oxide (DMPO) followed by brief treatment with the solid oxidant lead dioxide (PbO₂) yielded, after filtration, a cell-free solution that contained two nitroxyl adducts. The first was the hydroxyl radical adduct, 5,5-dimethyl-2-hydroxypyrrolidine-1-oxyl (DMPO-OH), which formed immediately upon PbO₂ oxidation. The second had a 6-line EPR spectrum typical of a carbon-centered radical (A_N=15.9 G; A_H=22.4 G) and formed more slowly. No radical signals were detected in the absence of either cells or PbO₂ treatment. The 6-line spectrum could be duplicated in model systems that contained ascorbate, DMPO and DMPO-OH, where the latter was formed from hydroxyl radicals generated by sonolysis or the cleavage of hydrogen peroxide with Fe²⁺ (Fenton reaction). In addition, enrichment of MC-1010 cells with ascorbate prior to spin trapping yielded the 6-line EPR spectrum as the principal adduct following PbO₂ oxidation and filtration. These results suggest that ascorbate reacted with DMPO-OH to form a carbon-centered ascorbyl radical that was subsequently trapped by DMPO. The requirement for mild oxidation to detect the hydroxyl radical adduct suggests that DMPO-OH formed in the cells was reduced to an EPR-silent form (i.e., the hydroxylamine derivative). Alternatively, the hydroxylamine derivative was the species initially formed. The evidence for endogenous hydroxyl radical formation in unstimulated leukocytes may be relevant to the leukemic nature of the MC-1010 cell line. The spin trapping of the ascorbyl radical is the first report of formation of the carbon-centered ascorbyl radical by means other than pulse radiolysis. Unless it is spin trapped, the carbon-centered ascorbyl radical immediately rearranges to the more stable oxygen-centered species that is passive to spin trapping and characterized by the well-known EPR doublet of A_H4=1.8 G.Abbreviation EPR Electron Paramagnetic Resonance     

Curcumin inhibits nitrogen dioxide induced oxidation of hemoglobin

Curcumin protects hemoglobin from nitrogen dioxide induced oxidation. Curcumin was also found to scavenge nitrogen dioxide in a concentration dependent way. The study also explains the ability of curcumin to protect hemoglobin from nitrite induced oxidation, where nitrogen dioxide is a key intermediate.     

大气NO2影响植物生长与代谢的研究进展

二氧化氮(NO₂)是大气氮氧化物之一,是大气气溶胶颗粒形成的主要成分,降低大气NO₂浓度可减轻空气中的雾霾.大气NO₂通过干沉降和湿沉降两种方式降落到植物叶片.植物吸收NO₂后主要通过两种代谢途径来降低空气中NO₂浓度: 一是主要在细胞质和叶绿体中利用还原酶的氮代谢途径,二是在质外体和细胞质中的歧化反应.植物吸收NO₂干扰了植物正常的生长和生理代谢,包括: 植物营养和生殖生长,植物体内硝酸还原酶(NaR)活性、亚硝酸还原酶(NiR)活性、氮素吸收、光合等生理代谢过程.对目前国内外有关大气NO₂影响植物生长与代谢的研究进展进行了综述,并对植物吸收NO₂的生理及分子机制的未来研究方向进行了展望.     

Change in fluxes of carbon dioxide,methane and nitrous oxide due to forest drainage of mire sites of different trophy

Northern peatlands accumulate atmospheric CO₂ thus counteracting climate warming. However, CH₄ which is more efficient as a greenhouse gas than CO₂, is produced in the anaerobic decomposition processes in peat. When peatlands are taken for forestry their water table is lowered by ditching. We studied long-term effects of lowered water table on the development of vegetation and the annual emissions of CO₂, CH₄ and N₂O in an ombrotrophic bog and in a minerotrophic fen in Finland. Reclamation of the peat sites for forestry had changed the composition and coverage of the field and ground layer species, and increased highly the growth of tree stand at the drained fen. In general, drainage increased the annual CO₂ emissions but the emissions were also affected by the natural fluctuations of water table. In contrast to CO₂, drainage had decreased the emissions of CH₄, the drained fen even consumed atmospheric CH₄. CO₂ and CH₄ emissions were higher in the virgin fen than in the virgin bog. There were no N₂O emissions from neither type of virgin sites. Drainage had, however, highly increased the N₂O emissions from the fen. The results suggest that post-drainage changes in gas fluxes depend on the trophy of the original mires.     

Kinetics of the reactions of nitrogen dioxide with glutathione,cysteine, and uric acid at physiological pH

Nitrogen dioxide (NO₂^•) is a key biological oxidant. It can be derived from peroxynitrite via the interaction of nitric oxide with superoxide, from nitrite with peroxidases, or from autoxidation of nitric oxide. In this study, submicromolar concentrations of NO₂^• were generated in < 1 μs using pulse radiolysis, and the kinetics of scavenging NO₂^• by glutathione, cysteine, or uric acid were monitored by spectrophotometry. The formation of the urate radical was observed directly, while the production of the oxidizing radical obtained on reaction of NO₂^• with the thiols (the thiyl radical) was monitored via oxidation of 2,2′-azino-bis-(3-ethylthiazoline-6-sulfonic acid). At pH 7.4, rate constants for reaction of NO₂^• with glutathione, cysteine, and urate were estimated as 2 × 10⁷, 5 × 10⁷, and 2 × 10⁷ M⁻¹ s⁻¹, respectively. The variation of these rate constants with pH indicated that thiolate reacted much faster than undissociated thiol. The dissociation of urate also accelerated reaction with NO₂^• at pH > 8. The thiyl radical from GSH reacted with urate with a rate constant of 3 × 10⁷ M⁻¹ s⁻¹. The implications of these values are: (i) the lifetime of NO₂^• in cytosol is < 10 μs; (ii) thiols are the dominant ‘sink’ for NO₂^• in cells/tissue, whereas urate is also a major scavenger in plasma; (iii) the diffusion distance of NO₂^• is 0.2 μm in the cytoplasm and < 0.8 μm in plasma; (iv) urate protects GSH against depletion on oxidative challenge from NO₂^•; and (v) reactions between NO₂^• and thiols/urate severely limit the likelihood of reaction of NO₂^• with NO• to form N₂O₃ in the cytoplasm.     

Ecological ramifications of the direct foliar uptake of nitrogen

The foliar incorporation of various reactive forms of nitrogen (N) has been identified and studied for nearly 30 years. However, the ecosystem-level ramifications of this uptake pathway have only recently been considered by the scientific community. In this review, I present our current understanding of the foliar uptake process and then discuss why this pathway of N addition to ecosystems should be considered separately from the bulk deposition of N to the soil surface. Direct foliar uptake is a direct addition of N to plant metabolism and could potentially more readily influence plant growth compared to soil-deposited N. Current ecosystem process models do not partition reactive N between foliar and soil entry pathways and the influence of N deposition on ecosystem C sequestration is likely inadequately represented in most models. I also outline several research priorities for the future understanding of the ecological consequences of foliar uptake of reactive N.     

Denitrification and nitrous oxide emissions from riparian forests soils exposed to prolonged nitrogen runoff

Compared to upland forests, riparian forest soils have greater potential to remove nitrate (NO₃) from agricultural runoff through denitrification. It is unclear, however, whether prolonged exposure of riparian soils to nitrogen (N) loading will affect the rate of denitrification and its end products. This research assesses the rate of denitrification and nitrous oxide (N₂O) emissions from riparian forest soils exposed to prolonged nutrient runoff from plant nurseries and compares these to similar forest soils not exposed to nutrient runoff. Nursery runoff also contains high levels of phosphate (PO₄). Since there are conflicting reports on the impact of PO₄ on the activity of denitrifying microbes, the impact of PO₄ on such activity was also investigated. Bulk and intact soil cores were collected from N-exposed and non-exposed forests to determine denitrification and N₂O emission rates, whereas denitrification potential was determined using soil slurries. Compared to the non-amended treatment, denitrification rate increased 2.7- and 3.4-fold when soil cores collected from both N-exposed and non-exposed sites were amended with 30 and 60 μg NO₃-N g⁻¹ soil, respectively. Net N₂O emissions were 1.5 and 1.7 times higher from the N-exposed sites compared to the non-exposed sites at 30 and 60 μg NO₃-N g⁻¹ soil amendment rates, respectively. Similarly, denitrification potential increased 17 times in response to addition of 15 μg NO₃-N g⁻¹ in soil slurries. The addition of PO₄ (5 μg PO₄-P g⁻¹) to soil slurries and intact cores did not affect denitrification rates. These observations suggest that prolonged N loading did not affect the denitrification potential of the riparian forest soils; however, it did result in higher N₂O emissions compared to emission rates from non-exposed forest soils.     

Spin Trapping of Free Radicals Produced in vivo in Heart and Liver During Endotoxemia

Studies using free radical scavengers and measurements of lipid peroxidation have suggested that free radicals are generated during endotoxemia. Conclusions from these studies have implied that free radicals may participate in the sequence of pathologic events following endotoxin challenge in the experimental animal. Current inferences of free radical generation and involvement have been derived from indirect evidence and are therefore inconclusive. To quantitate the generation of free radicals in vivo during endotoxemia this study employed the use of electron paramagnetic resonance spectroscopy (EPR) combined with spin trapping techniques. Five minutes before intraperitoneal endotoxin administration, trimethoxy-a-phenyl-t-butyl-nitrone [(MeO), PBN] was administered intraperitoneally. Experimental animals were always matched with control animals receiving no endotoxin. At either five minutes or twenty-five minutes following endotoxin administration animals were decapitated and hearts and livers were rapidly taken for lipid extraction and EPR evaluation. Analysis of the EPR spectra revealed hyperfine splitting constants that indicated the presence of carbon-centered radical spin adducts in both organ tissues from animals exposed to endotoxin for twenty-five minutes. No signals were present in hearts and livers taken five minutes after endotoxin administration. EPR evaluation did not indicate spin adduct formation in control tissue. These data directly demonstrate that activation of processes in vivo involving free radical generation occur early during endotoxemia, but are not detectable immediately after the endotoxin challenge.     

No significant nitrous oxide emissions during spring thaw under grazing and nitrogen addition in an alpine grassland

A recent study (Wolf et al., 2010) suggests that short—lived pulses of N₂O emission during spring thaw dominate the annual N₂O budget and that grazing decreases N₂O emissions during the spring thaw. To verify this we conducted year—round N₂O flux measurements from June 2010 to May 2011 in Tianshan alpine grassland in central Asia. No pulse emissions of N₂O were found at grazing management sites and nitrogen addition sites during the spring thaw. The contribution of the spring thaw to the total annual N₂O budget was small and accounted for only 6.6% of the annual fluxes, with winter emissions accounting for 16.7% and growing season emissions accounting for 76.7%. The difference in N₂O emissions attributable to grazing management was not significant (P > 0.05). Nitrogen input tended to increase N₂O emissions at N addition sites during the grass growing season compared with those at unfertilized sites. N₂O fluxes showed a significant correlation with air temperature and also with both soil temperature and soil water content at 10 cm depth.     

Exogenous sodium sulfide improves morphological and physiological responses of a hybrid Populus species to nitrogen dioxide

Gaseous nitrogen dioxide (NO₂) can disturb normal plant growth and trigger complex physiological responses. NO₂-induced responses are influenced by biotic or abiotic factors. In this study, we investigated the effects of exogenous sodium sulfide (Na₂S, 5 mmol L⁻¹) on epidermis and stomata related physico-chemical responses of hybrid poplar cuttings (Pouplus alba × P. berolinensis) to gaseous NO₂ (4 μl 1⁻¹) for three time periods (0, 14 and 48 h). We also investigated hydrogen sulfide (H₂S), nitrate-nitrogen and nitrate reductase activity (NR) in control and Na₂S treated plants. Our results showed that NO₂ exposure for 48 h led to the decline of NR, maximal PSII quantum yield (F_v/F_m), net photosynthetic rate (P_n), and dark respiration rate (R_d). The maximum rate for the post-illumination carbon dioxide burst (PIB) occurred in 48-h exposed leaves 13–15 s after darkening. Moreover, NO₂ exposure resulted in a significant increase in nitrogen percentage (from 0 to 33%) and a decrease in the macro and micro-elements of leaf surface. Spraying Na₂S aqueous solution on the leaf surfaces significantly increased the thicknesses of palisade/spongy tissue and H₂S content. Na₂S pretreatment alleviated NO₂-caused toxic effects as indicated by increased NR and higher values of P_n, F_v/F_m, and actual photochemical efficiency in light (ФPSII) compared with the control. Na₂S pretreatment had no significant impacts on PIB-based photorespiration or elements composition of a leaf surface.