A comparison of nutrient availability measured by chemical analysis and calculated from bioassay yields

The concentrations of soluble biologically available nitrogen and phosphorus were determined using Selenastrum capricornutum bioassays and compared with analytically measured soluble nitrate (NO₃-N) and soluble reactive phosphorus concentrations during enrichment studies in a South African impoundment. The NO₃-N analyses consistently underestimated the soluble biologically available nitrogen and the extent of the discrepancy decreased with increasing NO₃-N concentration. Biological availability of soluble organic nitrogen during the bioassays is suggested as a reason for the discrepancies. At low soluble reactive phosphorus concentrations the analytical measurements underestimated the soluble biologically available phosphorus while at high soluble reactive phosphorus concentrations the analytical measurements were considerable overestimates of soluble biologically available phosphorus. Possible reasons for the observed trend are discussed.     

Quantitative analysis of biomarkers,drugs and toxins in biological samples by immunoaffinity chromatography coupled to mass spectrometry or tandem mass spectrometry: A focused review of recent applications

Immunoaffinity chromatography (IAC), mass spectrometry and especially tandem mass spectrometry (MS/MS) represent the most efficient and reliable analytical techniques for specific isolation, unequivocal identification and accurate quantification of numerous natural and synthetic substances in biological samples. This review article focuses on the combined use of these outstanding methodologies in basic and clinical research and in life sciences for the quantitative analysis of low- and high-molecular mass biomarkers, drugs and toxins in urine, plasma or serum samples, in tissue and other biologicals systems published in the last decade. The analytes discussed in some detail include the biomarkers of oxidative stress 15(S)-8-iso-prostaglandin F_2α {15(S)-8-iso-PGF_2α} and 3-nitrotyrosine, the major urinary metabolite of the lipid mediators cysteinyl leukotrienes, i.e., the leukotriene E₄ (LTE₄), melatonin, and the major collagen type II neoepitope peptide in human urine.     

Quantitative trait locus analysis of circulating adhesion molecules in hyperlipidemic apolipoprotein E-deficient mice

Circulating soluble adhesion molecules have been suggested as useful markers to predict several clinical conditions such as atherosclerosis, type 2 diabetes, obesity, and hypertension. To determine genetic factors influencing plasma levels of soluble vascular cell adhesion molecule-1 (VCAM-1) and P-selectin, quantitative trait locus (QTL) analysis was performed on an intercross between C57BL/6J (B6) and C3H/HeJ (C3H) mouse strains deficient in apolipoprotein E-deficient (apoE^−/−). Female F₂ mice were fed a western diet for 12 weeks. One significant QTL, named sVcam1 (71 cM, LOD 3.9), on chromosome 9 and three suggestive QTLs on chromosomes 5, 13 and 15 were identified to affect soluble VCAM-1 levels. Soluble P-selectin levels were controlled by one significant QTL, named sSelp1 (8.5 cM, LOD 3.4), on chromosome 16 and two suggestive QTLs on chromosomes 10 and 13. Both adhesion molecules showed significant or an apparent trend of correlations with body weight, total cholesterol, and LDL/VLDL cholesterol levels in the F₂ population. These results indicate that plasma VCAM-1 and P-selectin levels are complex traits regulated by multiple genes, and this regulation is conferred, at least partially, by acting on body weight and lipid metabolism in hyperlipidemic apoE^−/− mice. Zuobiao Yuan and Zhiguang Su contributed equally.     

A copper‐specific microbial fuel cell biosensor based on riboflavin biosynthesis of engineered Escherichia coli

Copper pollution poses a serious threat to the aquatic environment; however, in situ analytical methods for copper monitoring are still scarce. In the current study, Escherichia coli Rosetta was genetically modified to express OprF and ribB with promoter P_t7 and P_cusC, respectively, which could synthesize porin and senses Cu²⁺ to produce riboflavin. The cell membrane permeability of this engineered strain was increased and its riboflavin production (1.45–3.56 μM) was positively correlated to Cu²⁺ (0–0.5 mM). The biosynthetic strain was then employed in microbial fuel cell (MFC) based biosensor. Under optimal operating parameters of pH 7.1 and 37°C, the maximum voltage (248, 295, 333, 352, and 407 mV) of the constructed MFC biosensor showed a linear correlation with Cu²⁺ concentration (0.1, 0.2, 0.3, 0.4, 0.5 mM, respectively; R² = 0.977). The continuous mode testing demonstrated that the MFC biosensor specifically senses Cu²⁺ with calculated detection limit of 28 μM, which conforms to the common Cu²⁺ safety standard (32 μM). The results obtained with the developed biosensor system were consistent with the existing analytical methods such as colorimetry, flame atomic absorption spectrometry, and inductively coupled plasma optical emission spectrometry. In conclusion, this MFC‐based biosensor overcomes the signal conversion and transmission problems of conventional approaches, providing a fast and economic analytical alternative for in situ monitoring of Cu²⁺ in water.     

Formation of 3-nitrotyrosine by riboflavin photosensitized oxidation of tyrosine in the presence of nitrite

The results of the present investigation show the susceptibility of tyrosine to undergo visible light-induced photomodification to 3-nitrotyrosine in the presence of nitrite and riboflavin, as sensitizer. By changing H₂O by D₂O, it could be established that singlet oxygen has a minor role in the reaction. The finding that nitration of tyrosine still occurred to a large extent under anaerobic conditions indicates that the process proceeds mainly through a type I mechanism, which involves the direct interaction of the excited state of riboflavin with tyrosine and nitrite to give tyrosyl radical and nitrogen dioxide radical, respectively. The tyrosyl radicals can either dimerize to yield 3,3′-dityrosine or combine with nitrogen dioxide radical to form 3-nitrotyrosine. The formation of 3-nitrotyrosine was found to increase with the concentration of nitrite added and was accompanied by a decrease in the recovery of 3,3′-dityrosine, suggesting that tyrosine nitration competes with dimerization reaction. The riboflavin photosensitizing reaction in the presence of nitrite was also able to induce nitration of tyrosine residues in proteins as revealed by the spectral changes at 430 nm, a characteristic absorbance of 3-nitrotyrosine, and by immunoreactivity using 3-nitrotyrosine antibodies. Since riboflavin and nitrite are both present endogenously in living organism, it is suggested that this pathway of tyrosine nitration may potentially occur in tissues and organs exposed to sunlight such as skin and eye.     

Iron interference in the quantification of nitrate in soil extracts and its effect on hypothesized abiotic immobilization of nitrate

Human alteration of the nitrogen cycle has stimulated research on nitrogen cycling in many aquatic and terrestrial ecosystems, where analyses of nitrate (NO₃ ⁻) by standard laboratory methods are common. A recent study by Colman et al. (Biogeochemistry 84:161–169, 2007) identified a potential analytical interference of soluble iron (Fe) with NO₃ ⁻ quantification by standard flow-injection analysis of soil extracts, and suggested that this interference may have led Dail et al. (Biogeochemistry 54:131–146, 2001) to make an erroneous assessment of abiotic nitrate immobilization in prior ¹⁵N pool dilution studies of Harvard Forest soils. In this paper, we reproduce the Fe interference problem systematically and show that it is likely related to dissolved, complexed-Fe interfering with the colorimetric analysis of NO₂ ⁻. We also show how standard additions of NO₃ ⁻ and NO₂ ⁻ to soil extracts at native dissolved Fe concentrations reveal when the Fe interference problem occurs, and permit the assessment of its significance for past, present, and future analyses. We demonstrate low soluble Fe concentrations and good recovery of standard additions of NO₃ ⁻ and NO₂ ⁻ in extracts of sterilized Harvard Forest soils. Hence, we maintain that rapid NO₃ ⁻ immobilization occurred in sterilized samples of the Harvard Forest O horizon in the study by Dail et al. (2001). Furthermore, additional evidence is accumulating in the literature for rapid disappearance of NO₃ ⁻ added to soils, suggesting that our observations were not the result of an isolated analytical artifact. The conditions for NO₃ ⁻ reduction are likely to be highly dependent on microsite properties, both in situ and in the laboratory. The so-called “ferrous wheel hypothesis” (Davidson et al., Glob Chang Biol 9:228–236, 2003) remains an unproven, viable explanation for published observations.     

Tyrosine nitration: Localisation,quantification, consequences for protein function and signal transduction

The nitration of free tyrosine or protein tyrosine residues generates 3-nitrotyrosine the detection of which has been utilised as a footprint for the in vivo formation of peroxynitrite and other reactive nitrogen species. The detection of 3-nitrotyrosine by analytical and immunological techniques has established that tyrosine nitration occurs under physiological conditions and levels increase in most disease states. This review provides an updated, comprehensive and detailed summary of the tissue, cellular and specific protein localisation of 3-nitrotyrosine and its quantification. The potential consequences of nitration to protein function and the pathogenesis of disease are also examined together with the possible effects of protein nitration on signal transduction pathways and on the metabolism of proteins.     

Kinetics of 3-nitrotyrosine modification on exposure to hypochlorous acid

Abstract

The markers 3-nitrotyrosine and 3-chlorotyrosine are measured as surrogates for reactive nitrogen species and hypochlorous acid respectively, which are both elevated in inflamed human tissues. Previous studies reported a loss of 3-nitrotyrosine when exposed to hypochlorous acid, suggesting that observations of 3-nitrotyrosine underestimate the presence of reactive nitrogen species in diseased tissue (Whiteman and Halliwell, Biochemical and Biophysical Research Communications, 258, 168–172 (1999)). This report evaluates the significance of 3-nitrotyrosine loss by measuring the kinetics of the reaction between 3-nitrotyrosine and hypochlorous acid. The results demonstrate that 3-nitrotyrosine is chlorinated by hypochlorous acid or chloramines to form 3-chloro-5-nitrotyrosine. As 3-nitrotyrosine from in vivo samples is usually found within proteins rather than as free amino acid, we also examined the reaction of 3-nitrotyrosine modification in the context of peptides. The chlorination of 3-nitrotyrosine in peptides was observed to occur up to 700-fold faster than control reactions using equivalent amino acid mixtures. These results further advance our understanding of tyrosine chlorination and the use of 3-nitrotyrosine formed in vivo as a biomarker of reactive nitrogen species.     

Site-specific Nitration of Apolipoprotein A-I at Tyrosine 166 Is Both Abundant within Human Atherosclerotic Plaque and Dysfunctional

We reported previously that apolipoprotein A-I (apoA-I) is oxidatively modified in the artery wall at tyrosine 166 (Tyr¹⁶⁶), serving as a preferred site for post-translational modification through nitration. Recent studies, however, question the extent and functional importance of apoA-I Tyr¹⁶⁶ nitration based upon studies of HDL-like particles recovered from atherosclerotic lesions. We developed a monoclonal antibody (mAb 4G11.2) that recognizes, in both free and HDL-bound forms, apoA-I harboring a 3-nitrotyrosine at position 166 apoA-I (NO₂-Tyr¹⁶⁶-apoA-I) to investigate the presence, distribution, and function of this modified apoA-I form in atherosclerotic and normal artery wall. We also developed recombinant apoA-I with site-specific 3-nitrotyrosine incorporation only at position 166 using an evolved orthogonal nitro-Tyr-aminoacyl-tRNA synthetase/tRNA_CUA pair for functional studies. Studies with mAb 4G11.2 showed that NO₂-Tyr¹⁶⁶-apoA-I was easily detected in atherosclerotic human coronary arteries and accounted for ∼8% of total apoA-I within the artery wall but was nearly undetectable (>100-fold less) in normal coronary arteries. Buoyant density ultracentrifugation analyses showed that NO₂-Tyr¹⁶⁶-apoA-I existed as a lipid-poor lipoprotein with <3% recovered within the HDL-like fraction (d = 1.063–1.21). NO₂-Tyr¹⁶⁶-apoA-I in plasma showed a similar distribution. Recovery of NO₂-Tyr¹⁶⁶-apoA-I using immobilized mAb 4G11.2 showed an apoA-I form with 88.1 ± 8.5% reduction in lecithin-cholesterol acyltransferase activity, a finding corroborated using a recombinant apoA-I specifically designed to include the unnatural amino acid exclusively at position 166. Thus, site-specific nitration of apoA-I at Tyr¹⁶⁶ is an abundant modification within the artery wall that results in selective functional impairments. Plasma levels of this modified apoA-I form may provide insights into a pathophysiological process within the diseased artery wall.     

Chemical Bonding and Physical Trapping of Sulfur in Mesoporous Magnéli Ti4O7 Microspheres for High‐Performance Li–S Battery

Various host materials have been investigated to address the intrinsic drawbacks of lithium sulfur batteries, such as the low electronic conductivity of sulfur and inevitable decay in capacity during cycling. Besides the widely investigated carbonaceous materials, metal oxides have drawn much attention because they form strong chemical bonds with the soluble lithium polysulfides. Here, mesoporous Magnéli Ti₄O₇ microspheres are prepared via an in situ carbothermal reduction that exhibit interconnected mesopores (20.4 nm), large pore volume (0.39 cm³ g^?1), and high surface area (197.2 m² g^?1). When the sulfur cathode is embedded in a matrix of mesoporous Magnéli Ti₄O₇ microspheres, it exhibits a superior reversible capacity of 1317.6 mA h g^?1 at moderate current (C/10) and a low decay in capacity of 12% after 400 cycles at C/5. Strong chemical bonding of the lithium polysulfides to Ti₄O₇, as well as effective physical trapping in the mesopores and voids in the matrix are considered responsible for the improved electrochemical performance. A mechanism of the physical and chemical interactions between mesoporous Magnéli Ti₄O₇ microspheres and sulfur is proposed based on systematic investigations.     

Twoplex 12/13C6 aniline stable isotope and linkage‐specific sialic acid labeling 2D‐LC‐MS workflow for quantitative N‐glycomics

Quantitative glycomics represents an actively expanding research field ranging from the discovery of disease‐associated glycan alterations to the quantitative characterization of N‐glycans on therapeutic proteins. Commonly used analytical platforms for comparative relative quantitation of complex glycan samples include MALDI‐TOF‐MS or chromatographic glycan profiling with subsequent data alignment and statistical evaluation. Limitations of such approaches include run‐to‐run technical variation and the potential introduction of subjectivity during data processing. Here, we introduce an offline 2D LC‐MS^E workflow for the fractionation and relative quantitation of twoplex isotopically labeled N‐linked oligosaccharides using neutral ¹²C₆ and ¹³C₆ aniline (Δmass = 6 Da). Additional linkage‐specific derivatization of sialic acids using 4‐(4,6‐dimethoxy‐1,3,5‐trizain‐2‐yl)‐4‐methylmorpholinium chloride offered simultaneous and advanced in‐depth structural characterization. The potential of the method was demonstrated for the differential analysis of structurally defined N‐glycans released from serum proteins of patients diagnosed with various stages of colorectal cancer. The described twoplex ¹²C₆/¹³C₆ aniline 2D LC‐MS platform is ideally suited for differential glycomic analysis of structurally complex N‐glycan pools due to combination and analysis of samples in a single LC‐MS injection and the associated minimization in technical variation.     

CX3CR1 Is Expressed by Human B Lymphocytes and Meditates CX3CL1 Driven Chemotaxis of Tonsil Centrocytes

Background

Fractalkine/CX₃CL1, a surface chemokine, binds to CX₃CR1 expressed by different lymphocyte subsets. Since CX₃CL1 has been detected in the germinal centres of secondary lymphoid tissue, in this study we have investigated CX₃CR1 expression and function in human naïve, germinal centre and memory B cells isolated from tonsil or peripheral blood.

Methodology/Principal Findings

We demonstrate unambiguously that highly purified human B cells from tonsil and peripheral blood expressed CX₃CR1 at mRNA and protein levels as assessed by quantitative PCR, flow cytometry and competition binding assays. In particular, naïve, germinal centre and memory B cells expressed CX₃CR1 but only germinal centre B cells were attracted by soluble CX₃CL1 in a transwell assay. CX₃CL1 signalling in germinal centre B cells involved PI3K, Erk1/2, p38, and Src phosphorylation, as assessed by Western blot experiments. CX₃CR1⁺ germinal centre B cells were devoid of centroblasts and enriched for centrocytes that migrated to soluble CX₃CL1. ELISA assay showed that soluble CX₃CL1 was secreted constitutively by follicular dendritic cells and T follicular helper cells, two cell populations homing in the germinal centre light zone as centrocytes. At variance with that observed in humans, soluble CX₃CL1 did not attract spleen B cells from wild type mice. OVA immunized CX₃CR1⁻/⁻ or CX₃CL1⁻/⁻ mice showed significantly decreased specific IgG production compared to wild type mice.

Conclusion/Significance

We propose a model whereby human follicular dendritic cells and T follicular helper cells release in the light zone of germinal centre soluble CX₃CL1 that attracts centrocytes. The functional implications of these results warrant further investigation.     

Potential Sources of Error in the Measurement of Low Rates of Planktonic Photosynthesis and Excretion

THE introduction of liquid scintillation counting techniques to the study of aquatic productivity^1–3 has stimulated interest in the accurate determination of low rates of algal photosynthesis, such as occur in tropical waters or in deep living phytoplankton populations. Attention is also being directed to the measurement of soluble extracellular products of photosynthesis⁴. Our studies have revealed some potential sources of interference in the widely used ¹⁴C technique when used to study low rates: (1) retention of NaH¹⁴C0₃ by filters; (2) passive adsorption of radioactivity by filters and/or seston and (3) soluble and insoluble radioactive impurities in commercially available NaH¹⁴C0₃.     

Luminescent and hydrophilic LaF3–polymer nanocomposite for DNA detection

Luminescent LaF₃–Ce³⁺/Tb³⁺ nanocrystals have been successfully prepared via a simple wet chemical technique. For the next bioapplication, these nanoparticles dispersed in cyclohexane have also been functionalized with poly(St‐co‐MAA), based on a designed oil‐in‐water microemulsion system. These polymer‐coated nanospheres are water‐soluble and bioconjugable. Unlike semiconductor quantum dots, the as‐prepared lanthanum fluoride nanocrystals possess non‐size‐dependent emissions and completely stable photocycles. With functionalized LaF₃ nanospheres as fluorescence probes, a fluorescence method was developed for the rapid quantitative analysis of DNA, due to the quenching effect of fluorescence by the DNA. Under optimum conditions, the fluorescence intensity was proportional to the concentration of the introduced DNA over the range 2.5–35 µg/mL for calf thymus DNA (ctDNA) and 2.5–30 µg/mL for fish sperm DNA (fsDNA), respectively. Copyright © 2008 John Wiley & Sons, Ltd.     

Translocation and Intracellular Distribution of Tritiated Gibberellin A3

The localization of tritium-radioactivity in dwarf kidney bean plants (Phaseolus vulgaris) of ³H-gibberellm A₃(³H-GA₃) applied in a large quantity was investigated in advance of the study on GA₃ metabolism in this plant. Immediately after the application of ³H-GA₃, the radioactivity was distributed uniformly in the top of this plant; no further transportation of the radioactivity into the growing apical region from mature leaves and stems was the observed as the growth stage proceeded. An investigation on the intracellular localization of the radioactivity demonstrated that most part of the radioactivity was found in the cellular soluble fraction, while no radioactivity was detected in such subcellular particles as nuclei, mitochondria and microsomes. Examinations of the occurrence of GA₃ bound with such macromolecules as RNA and protein gave negative results.     

Organization,replication and modification of the human genome: Differential methylation of two classes of HeLa nuclear DNA separated on Ag+−Cs2SO4 gradients

HeLa nuclear DNA sediments as a single peak, in neutral CsCl, while it is separated in a heavier and a lighter components, in alkaline Ag⁺–Cs₂SO₄. The heavy fraction, on the average, represents about 20% of the total DNA. CsCl analytical ultracentrifugation shows that heavy DNA bands at 1.715 g/cm³ and contains 53% GC (10% of the total GC), whereas light DNA bands at 1.703 g/cm³ and contains 40% GC (32% of the total GC). Coherently, Tm values in 0.1 x SSC are 82.5°C, for heavy DNA, and 72.5°C, for light DNA. After treatment with [³H-methyl-S-adenosyl-L-methionine in isolated nuclei, the concentration of labelled 5-methylcytosine was found to be highest in the more dense regions of the heavy peak and in the less dense regions of the light peak. Exposure to ultrasound modifies the quantitative relationship of the two peaks and improves the separation of supermethylated AT- and GC-rich DNAs. Four possible triplets as sites for DNA-methylase recognition are discussed.     

Nitric oxide and peroxynitrite. The ugly,the uglier and the not so good

Nitric oxide, a gaseous free radical, is poorly reactive with most biomolecules but highly reactive with other free radicals. Its ability to scavenge peroxyl and other damaging radicals may make it an important antioxidant in vivo, particular in the cardiovascular system, although this ability has been somewhat eclipsed in the literature by a focus on the toxicity of peroxynitrite, generated by reaction of O^·-₂ with NO^· (or of NO^- with O₂). On balance, experimental and theoretical data support the view that ONOO^- can lead to hydroxyl radical (OH^·) generation at pH 7.4, but it seems unlikely that OH^· contributes much to the cytotoxicity of ONOO^-. The cytotoxicity of ONOO^- may have been over-emphasized: its formation and rapid reaction with antioxidants may provide a mechanism of using NO^· to dispose of excess O^·-₂, or even of using O^·-₂ to dispose of excess NO^·, in order to maintain the correct balance between these radicals in vivo. Injection or instillation of “bolus” ONOO^- into animals has produced tissue injury, however, although more experiments generating ONOO^- at steady rates in vivo are required. The presence of 3-nitrotyrosine in tissues is still frequently taken as evidence of ONOO^- generation in vivo, but abundant evidence now exists to support the view that it is a biomarker of several “reactive nitrogen species”. Another under-addressed problem is the reliability of assays used to detect and measure 3-nitrotyrosine in tissues and body fluids: immunostaining results vary between laboratories and simple HPLC methods are susceptible to artefacts. Exposure of biological material to low pH (e.g. during acidic hydrolysis to liberate nitrotyrosine from proteins) or to H₂O₂ might cause artefactual generation of nitrotyrosine from NO^-₂ in the samples. This may be the origin of some of the very large values for tissue nitrotyrosine levels quoted in the literature. Nitrous acid causes not only tyrosine nitration but also DNA base deamination at low pH: these events are relevant to the human stomach since saliva and many foods are rich in nitrite. Several plant phenolics inhibit nitration and deamination in vitro, an effect that could conceivably contribute to their protective effects against gastric cancer development.     

Inositol Trisphosphate Metabolism in Carrot (Daucus carota L.) Cells

The metabolism of exogenously added d-myo-[1-³H]inositol 1,4,5-trisphosphate (IP₃) has been examined in microsomal membrane and soluble fractions of carrot (Daucus carota L.) cells grown in suspension culture. When [³H]IP₃ was added to a microsomal membrane fraction, [³H]IP₂ was the primary metabolite consisting of approximately 83% of the total recovered [³H] by paper electrophoresis. [³H]IP was only 6% of the [³H] recovered, and 10% of the [³H]IP₃ was not further metabolized. In contrast, when [³H]IP₃ was added to the soluble fraction, approximately equal amounts of [³H]IP₂ and [³H]IP were recovered. Ca²⁺ (100 micromolar) tended to enhance IP₃ dephosphorylation but inhibited the IP₂ dephosphorylation in the soluble fraction by about 20%. MoO₄²⁻ (1 millimolar) inhibited the dephosphorylation of IP₃ by the microsomal fraction and the dephosphorylation of IP₂ by the soluble fraction. MoO₄²⁻, however, did not inhibit the dephosphorylation of IP₃ by the soluble fraction. Li⁺ (10 and 50 millimolar) had no effect on IP₃ metabolism in either the soluble or membrane fraction; however, Li⁺ (50 millimolar) inhibited IP₂ dephosphorylation in the soluble fraction about 25%.     

The Effect of Lipopolysaccharide from <Emphasis Type="Italic">Proteus mirabilis</Emphasis> on the Level of the Stable End Metabolic Products of Nitric Oxide in Blood Platelets

Nitric oxide (^•NO) plays an important role in a number of physiologic processes. Evidence exists that ^•NO, which stimulates soluble guanylate cyclase and enhances cyclic guanosine monophosphate (cGMP) levels, may inhibit platelet activation. In contrast, during platelet activation induced by different agonists, synthesis of ^•NO in platelets occurs. In these studies, production of the stable end-products of ^•NO-nitrite and nitrate (NO_x) in human platelets, stimulated by different doses of lipopolysaccharide from Proteus mirabilis (LPS; endotoxin), has been evaluated. LPS is a weak platelet agonist that may activate various steps of platelet activation with the generation of reactive oxygen species. The mechanism of platelet activation induced by the endotoxin is not known. The aim of the present study was to measure the level of nitrite and NO_x in blood platelets treated with LPS and to examine the level of nitrotyrosine in platelet proteins caused by LPS. Our results show that LPS at a low concentration (6.8 ng/ml) caused a decrease (approximately 80%) in the NO_x level, whereas at higher concentrations (13.6 and 25 ng/ml) it induced an increase in the NO_x level (approximately 210% and 260%, respectively). Our results indicate that LPS, like other agonists (thrombin, platelet-activating factor), can stimulate ^•NO production in platelets. After incubating platelets with LPS, we also observed a distinct increase in platelet protein nitration (3-nitrotyrosine).     

Reassessing the nitrogen relations of Arctic plants: a mini-review

The Arctic is often assumed to be an NH₄⁺-dominated ecosystem. This review assesses the validity of this assumption. It also addresses the question of whether Arctic plant growth is limited by the ability to use the forms of nitrogen that are available. The review demonstrates that several sources of soil nitrogen are available to Arctic plants, including soluble organic nitrogen (e.g. glycine, aspartic acid and glutamic acid), NH₄⁺ and NO^?₃. In mesic Arctic soils, soluble organic nitrogen is potentially more important than either NH⁺₄ or NO^?₃. Many Arctic species are capable of taking up soluble organic nitrogen (either directly and/or in association with ectomycorrhizae), with the greatest potential for soluble organic nitrogen uptake being exhibited by deciduous species. The ability to take up soluble organic nitrogen may enable some Arctic plants to avoid nitrogen limitations imposed by the slow rate of organic matter decomposition. NO^?₃ is also present in many Arctic soils, especially calcareous soils and soils near flowing water, animal burrows and bird cliffs. Arctic species characteristic of mesic and xeric habitats are capable of taking up and assimilating NO^?₃. Even when present in lower concentrations in soils than NH⁺₄, NO^?₃ is still an important source of nitrogen for some Arctic plants. Arctic-plants therefore have a variety of nitrogen sources available to them, and are capable of using those nitrogen sources. Taken together, these findings demonstrate that the Arctic is not an NH⁺₄dominated ecosystem. Symbiotic fixation of atmospheric N₂ does not appear to be an important source of nitrogen for Arctic plants. The reliance of Arctic plants on internal recycling of nitrogen substantially reduces their dependence on soil nitrogen uptake (this is particularly the case for slow-growing evergreens). Despite the high level of internal nitrogen recycling, Arctic plant growth remains limited by the low levels of available soil nitrogen. However, Arctic plant growth is not limited by an inability to utilize any of the available forms of nitrogen. The potential effects of climatic warming on nitrogen availability and use are discussed. The question of whether the Arctic ecosystem is uniquely different from temperate nitrogen-deficient ecosystems is also assessed.